CA2190761A1 - Novel plants and processes for obtaining them - Google Patents

Novel plants and processes for obtaining them

Info

Publication number
CA2190761A1
CA2190761A1 CA002190761A CA2190761A CA2190761A1 CA 2190761 A1 CA2190761 A1 CA 2190761A1 CA 002190761 A CA002190761 A CA 002190761A CA 2190761 A CA2190761 A CA 2190761A CA 2190761 A1 CA2190761 A1 CA 2190761A1
Authority
CA
Canada
Prior art keywords
starch
plant
mutant
grain
genotype
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
CA002190761A
Other languages
French (fr)
Inventor
Peter Lewis Keeling
Frances Katz
Ming-Tang Chang
Richard Hauber
Robert Friedman
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Syngenta Ltd
American Maize-Products Co
Original Assignee
Peter Lewis Keeling
Frances Katz
Ming-Tang Chang
Richard Hauber
Robert Friedman
Zeneca Limited
American Maize-Products Company
Syngenta Limited
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to US26392194A priority Critical
Priority to US08/263,921 priority
Priority to US34660294A priority
Priority to US08/346,602 priority
Priority to US08/487,466 priority
Priority to US47406395A priority
Priority to US08/487,466 priority patent/US5576048A/en
Priority to US08/474,063 priority
Application filed by Peter Lewis Keeling, Frances Katz, Ming-Tang Chang, Richard Hauber, Robert Friedman, Zeneca Limited, American Maize-Products Company, Syngenta Limited filed Critical Peter Lewis Keeling
Publication of CA2190761A1 publication Critical patent/CA2190761A1/en
Application status is Abandoned legal-status Critical

Links

Classifications

    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23GCOCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G1/00Cocoa; Cocoa products, e.g. chocolate; Substitutes therefor
    • A23G1/30Cocoa products, e.g. chocolate; Substitutes therefor
    • A23G1/32Cocoa products, e.g. chocolate; Substitutes therefor characterised by the composition containing organic or inorganic compounds
    • A23G1/42Cocoa products, e.g. chocolate; Substitutes therefor characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
    • A23G1/423Cocoa products, e.g. chocolate; Substitutes therefor characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing microorganisms, enzymes
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/10Seeds
    • AHUMAN NECESSITIES
    • A21BAKING; EDIBLE DOUGHS
    • A21DTREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
    • A21D2/00Treatment of flour or dough by adding materials thereto before or during baking
    • A21D2/08Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
    • A21D2/14Organic oxygen compounds
    • A21D2/18Carbohydrates
    • A21D2/186Starches; Derivatives thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23GCOCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G1/00Cocoa; Cocoa products, e.g. chocolate; Substitutes therefor
    • A23G1/30Cocoa products, e.g. chocolate; Substitutes therefor
    • A23G1/56Cocoa products, e.g. chocolate; Substitutes therefor making liquid products, e.g. for making chocolate milk drinks and the products for their preparation, pastes for spreading, milk crumb
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23GCOCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G3/00Sweetmeats; Confectionery; Marzipan; Coated or filled products
    • A23G3/34Sweetmeats, confectionery or marzipan; Processes for the preparation thereof
    • A23G3/346Finished or semi-finished products in the form of powders, paste or liquids
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23GCOCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G3/00Sweetmeats; Confectionery; Marzipan; Coated or filled products
    • A23G3/34Sweetmeats, confectionery or marzipan; Processes for the preparation thereof
    • A23G3/36Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds
    • A23G3/364Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
    • A23G3/366Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins containing microorganisms, enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A23B - A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/20Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents
    • A23L29/206Foods or foodstuffs containing additives; Preparation or treatment thereof containing gelling or thickening agents of vegetable origin
    • A23L29/212Starch; Modified starch; Starch derivatives, e.g. esters or ethers
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C12N15/8245Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; THEIR TREATMENT, NOT COVERED BY OTHER CLASSES
    • A23GCOCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
    • A23G2200/00COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF containing organic compounds, e.g. synthetic flavouring agents
    • A23G2200/06COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF containing organic compounds, e.g. synthetic flavouring agents containing beet sugar or cane sugar if specifically mentioned or containing other carbohydrates, e.g. starches, gums, alcohol sugar, polysaccharides, dextrin or containing high or low amount of carbohydrate

Abstract

The invention relates to a transgenic or mutated plant having genomic material which alters the normal starch synthesis pathway within the plant. More specifically, the present invention relates to a plant having a genotype which creates new forms of starch in significant quantity. Particularly, the invention relates to grain having an embryo with a genotype heterozygous for two or more wild type genes (for example, Aa/Bb) and an endosperm having a genotype heterozygous for such genes (for example, AAa/BBb or AAa/bbB or aaA/BBb or aaA/bbB) and the starch produced therefrom.

Description

WO 9513~026 2 1 9 0 7 6 1 PCr~llS95J0~828 NOVEL PLANTS AND PROCESSES FOR OBTAINING THEM
FIELD OF THE ~VENlION
5 The invention relates to a transgenic or mutated plant having b~enomic maoerial which alters the normal starch synthesis pathway within the plant. More specifically, the present invention relates to a plant having a genotype which creates new forms of starch in significant quantity. Particularly, the invention relates to grain having an embryo with a genotype h~ u~bu.,.. for two or more wild type genes (for example,10 Aa/Bb) and an endosperm having a genotype ll~,t~,.uL~bu.~. for such genes (for example, AAa/BBb or AAa/bbB or aaA/BBb or aaA/bbB) and the starch produced therefrom.
Such grain are produced by pollinating a plant having a genotype llul~u~ ~bU..S
15 recessive for at least one gene and wild type for another gene (for example, aalBB) with pollen from another plant having a genotype llullluL~buuD recessive for at least one other gene and wild type for the other gene (for example, AA~bb).
BACKGROUND OF THE ~IVEN~ON
Most plants produce and store starch. These plants have a starch synthesis pathway for starch prûduction. The arnount of starch produced varies with the type of plant. The most commonly known starch producing plants are the cereal grains. These cereals include rice, maize, sorghum, barley, wheat, rye, and oats. Additionally, the 25 potato family, including the sweet potatoes and certain fruits, like the banana, are known as starch producing.
Starch is an impor~ant end-product of carbon fixation during ~Jl.vlu~ ' in leaves and is an important storage product in seeds and fruits. In economic oerrns, the 3 0 starch produced by the edible portions of three grain crops, wheat, rice and maize, provide ~ , two-thirds of the world's food calculaoed as calories.
I

SUBSTITUT~ SHEET (FiUL~ 26 W09513~026 p~"~ c ~

2 1 ~076 1 Starch from plants is used in various ways. For examp~e, it can be extracted and used for cooking and food processing. Starch can be left in the grain or plant and 35 used for animal and human . Starch can also be used in the distillation process for processing alcohols, for example, starch can be converted into ethanol.
Additional starch can convert to high-fructose syrup and other industrial Starch is defined in the dictionary as a granular solid which is chemically a 40 complex carbohydrate which is used in adhesives, sizes, foods, cosmetics, medicine, etc. More generally, starch is comprised of amylose and ~ y., Amylose and lu~ is synthesized in the plastid ,:, t...~,.-L (the chloroplast in 1' , ' cells or the amyloplast in non-~,l.v~u,y,.~..,Li., cells). Different plants generate differing proportions of ~".~, and amylose. r. , the different 45 branching pattems of ~..,!u~li,, and different chain lengths of amylose and ~ lu~lill chains gives rise to different starch propefies~ Thus, the fine structure of amy]ose and o~ lu~ill is different in different plants so that the branching patterns and ~ vaty UUllSl~ld~ resulting in new and novel properties which are useful in different 1." Until now there have been four ways of making 50 starches with special properties: (i) using statches extracted ftom different plant species, (ii) using statches extracted from mutant lines of palticular plants, (iii) using natutal and mutant starches which had been chemically modified, and (iv) using natural and mutant statches which had been physically modified. In all cases the new statches were valuable because of the special properties provided for by the new starch type.
It is known that mutant genes in plants affect the propetties of the statch. A
variety of statch related mutant genes in maize have been identified and some have been cled. These mutant genes were named accotding to the physical appeatance (phenotype) of the mai~e ketnel ot the properlies of the statch. These tecessive mutant 60 genes include waxy (wx), sugaty (su) [which includes but is not limited to sugary-l (sul), sugary-2 (su2), sugaty-3 (su3), sugary4 (su4)] dull (du), amylose extender (ae), SUBSTITUT~ Sl iEET (RULE ~6) WO 9~/35026 ~ 1 ~ 0 7 6 1 PCTIUS95107828 homy (h), shrunken (sh) which includes, but is not limited to, shrunken-l (sh-l), shrunken-2 (sh-2). Some of these recessive gene mutarts produce an isoform of a known enzyme in the starch synthesis pathway. The recessive mutant alleles of these 65 genes result in a complete or nearly complete reduction in the activity of a specific isoform of one enzyme (hereinafter defined as complete reduction of enzyme isoform activity) in the pathway when IIUIIIU~6V..D in a plarlt or when expressed in sufficient levels in a transgenic plant. This change in the starch synthesis pathway causes the fommation of starches with different properties, Several crop varieties are known which produce different types of starch. The type of quality of starch makes it suitable for certain purposes, including particular methods of processing or particular end-uses. Naturally-occurring maize mutants produce starches of differing fine structure suitable for use in various food products and 75 other l A]though known mutants produce altered starch, some of these lines are not suitable for crop breeding and/or for the farmers' purposes. For example, they can give relatively poor yields, and/or are difficult to process and/or carl have poor . .
In order to generate different starches, single and double mutant plarlts have been bred. A single mutant is a plant that is IIUIIIV~6~JL.D for one recessive mutant gene. For example, waxy maize, waxy rice, waxy barley, and waxy sorghum have thehu~llv~y~v~D mutant waxy (wx) gene. Whilst starches from waxy genotypes have very little or no amylose, another mutation known as amylose extender (ae) results in starch 85 with high amylose. A double mutant is a single plant that has t~ v~ .,., (or full expression) of two recessive mutant genes. For example, the wxfll double mutant is taught in U.S. patent 4,789,738. Many other novel starches have been provided in other starch patents in which double or triple mutants are generated (for example, US Patent Serial Numbers: 4789557, 4790997, 4774328, 4770710, 4798735, 4767849, 4801470, 90 4789738, 4792458 and 5009911 which describe naturally-occurfiing maize mutants producing starches of differing fine structure suitable for use in various food products).

SUBSTlTvTE SHEET (RULE 26) wo ss/3so26 2 1 9 ~ 7 6 1 PCT/IIS95/07U8 The present invention is highly surprising in light of these ~ because it produces altered starch and does not require double or triple mutants.
Normal starch is defined as starch which is not chemically modified (by peop1e) or which is produced from a plant that has the expected genes (wild type) regulating the starch synthesis pathway. For ease of reading, double lower-case letters, for example aa, shall refer to a liuil~u~6v~o recessive mutant gene, double upper case letters, for example AA, shall refer to a l1UIIIU~ D nûn-mutant gene (wild type), and 100 one upper-case and one lowercase letter, for example Aa, shall refer to a non-I~UIIIU~A~ o set of genes, one mutant, one non-mutant. Different letters in the same size shall mean different genes; "aalbb" would be a double mutant; "aalbB" would be a single homozygous mutant gene and a l.~,t~,.u~gu.,. mutant gene in the genome of the plant. For purposes of this application, the order of any three letters on one side of the 105 slash can be , ' , ' and will not define the parent that donated the gene. For example, AAa~bbB is defined to be equivalent to aAA/bBb, AaA/Bbb, AaAlbBb, aAA/Bbb, and the like.
Although maize plants and the embryo are diploid, maize endosperm is triploid.
110 The endosperm genotype has two gene doses which are inherited from the fernale plant portion and one gene dose which is inherited from the pollen or male plantportion. Thus, if a single mutant plant "aa" is used as the female and crossed to a non-mutant plant "AA" male, then the endospemm in the kemel of this female plant would be "aaA". If a non-mutant plant "AA" is crossed to a mutant plant "aa" with the non-115 mutant as the female, the endospetm on the kemel of the female plant will be "Aaa",because two gene doses come from the female and one from the male plant. Classic teæhing is that the mutant gene is recessive and the non-mutant is dominant; therefore, the starch produced by a plant having the following gene doses in the endospemm "aaA"
or "AAA" or "AAa" results in the nommal starch in the expected amounts. However,120 the endosperm of a IIUIIIUL~6~..0 mutant plant "aa" æting as the female crossed to a l~u~u~ o mutant plant "aa" acting as the male plant results in the endospemm having SUBSTITUT~ SHEET (RULE 26) ~ .

WO 95135026 r~

the gene dosage "aaa". This endosperm gives starch with different properties from normal starch. Likewise, the starch from a double mutant having of an endosperm which is "aaalbbb" shows differences in starch properties from normal starch. These 12 5 starch differences are useful in that they can replace chemically modified starches or be used with or in foodstuffs or as grain in alcohol production or in general starch industria] ~
Clearly, it appears that production of grain having starch with different physical 13 0 properties of starch requires the crossing of two mutated plants to generate grain which ;S IIUIIIUL~ recessive for both genes. Mutant plants are less predictable than standard plants.
There is a recurrent problem with the production of grain and extraction of 135 starch from double mutant hybrids and/or inbreds and some single mutants. The amount of starch produced is usually less than the amount of starch produced by the non-mutant plant, there is also a loss in starch granule size and/or starch granule integrity. This problem with known double mutant lines which produce !
altered starch in which the quantity of starch produced in the crop is relatively low can 140 result in poor g ' ".y of the seed. r~.tl,~.",u.~, the reduced starch yield of the seed appears to be I ~ui.ld~l~ since the mutations cause the normal starch synthesis ~ of the cells to be disrupted. There remains a need for a way to produce grain having structurally altered starch structurally altered starch or altered properties without a significant loss of yield or reduced starch granule size or integrity.145 SUMMARY OF TE~ INVEN~ON
A first object of the present invention is to provide a method of developing - hybrid plants having altered complex ~uI,vh~.' content of the grain which does not 15 0 require the crossing of double mutant inbreds.

SUBSrlTUTE SHEET (RULE 26) W095/35026 2 ~ ~Q76 t PCI/US95/07828 An object of the present invention is to provide plants that produce grain having altered starch properties.
155 Another object of the present invention is to provide transgenic plants that produce grain having altered starch properties.
Yet another object of the present invention is to provide a m~ize plant that produces both altered starch and larger quantities of starch than the associated mutant 16 0 plants produce.
Still another object of the present invention is to provide new plants which contain genes which produce incomplete reduction of the activity of at least twoisoforms of the specific enzymes in the starch synthesis pathway of the plant.

A further object of the present invention is to provide maize plants which have with the genotype "AAalBBb or AAalbbB or aaAlBBb or aaA/bbB".
Yet another object of the prvsent invention is a plant producirlg the following 17 0 endosperm genotype "wxwxWX/AeAeae".
Still an additional object of the present invention is to provide the altered starch which can be produced by plants having the genotype "AAalBBb or AAalbbB or aaA/BBb or aaA/bbB".

Yet another object of the present invention is to provide novel uses of the starch obtained from the Maize plants of the present invention.
The present invention broadly covers a method of producing 180 generally which have the endosperm genotype of AAa/bbB and certain including endosperm which is waxy, waxy, amylose extender (wxwxWx/AEAEae).

SUBSTITUT~ SHEET (RULE 26) The method of producing grain with altered starch qualities includes the steps of planting the female acting parent which is capable of flowering. The female parent having ' ly complete reduction of at least one specific isoform enzyme in the 185 starch synthesis pathway. This can be by a bul~u~,uu~ recessive mutant or by the partial down regulation of the wild type gene through the use of a cloned gene using techniques generally known as antisense or co ~ ' or sense-down regulation.
Additionally the female has incomplete reduction of at least one specific isofoTm enzyme in starch synthesis pathway. This can be by a l.~.~.u~,~.~ recessive mutant 190 gene or partial down regulation. Regardless of how the female is produced it should only act as the female portion. To assure this a step includes eliminating the first parent's capability to produce pollen. The method includes the step of pollinating the female acting parent with the pollen of the male acting parent which is a non-mutant parent. Harvesting the grain produced by said first parent. Additionally, the method can 195 include the extraction of starch from the grain.
This invention also ~ a plant having genomic material which includes genes which give incomplete reduction of the activity of at least two specific isoforms of the enzymes in the starch synthesis pathway of said planL And the starch it 200 produces which has altered structure when compared with the starch formed by a similar plant as described but which comprises genomic material which does not form isoforms of the enzymes in the starch synthesis pathway of the plant.
A plant which forms said starch in grain such as cereal grains. Grain produced 205 by a female plant having for example a waxy genotype (wxwx) crossed with a male plant having for example an amylose extender genotype (aeae) in which the genotype of the endosperm of the gr~un is wxwxWx/AeAeae.
In other words, the present invention is a starch producmg plant comprising 210 genomic material which includes genes which give incomplete reduction of the activity of at least two specific isoforms enzymes in the starch synthesis pathway of said plant SUBSTITUTE SHEET (RULE 26) wo ss/3so26 ~ r~ /n~s whereby said plant produces ! ' ' " 1~/ more starch than said plant would produce if said genes gave complete reduction of the activity of the same two specific isoforms of the enzymes within the starch synthesis pathway.

Grain having an endosperm that has two genes which contain one gene dose of recessive mutant gene and two doses of wild type; and, having two gene doses of recessive mutant and one dose of wild type. Within this description the invention r~ grain having endosperm genotypes of wxwxWx/AeAeae, or 220 aeaeAe/WxWxwx, or wxwxWx/DuDudu, or duduDu/WxWxwx, or aeæAe/DuDudu, or duduDu/AeAeae, or wxwxWx/SuSusu, or susuSu/WxWxwx, or æaeAe/SuSusu, or susuSu/AeAeæ, or duduDu/SuSusu, or susuSu/DuDudu and the like.
The starch from a grain having a genotype of wxwxWx/AeAeae. The starch 22 5 from a grain having a genotype of A= /WxWxwx.
A female plant having a diploid genotype of aa/BB and having a hiploid genotype of aaA/BBb where a is a recessive mutant gene and A is the wild type gene, and b is a recessive mut~nt gene and B is the wild type gene such that the starch is 2 3 0 altered from the normal starch where a and b can be selected from ae, wx, sh, bt, h, su, fl, op and B and A can be selected from Ae, Wx, Sh, Bt, H, Su, rl, Op.
The starch obtained in accordance with the preænt invention produces a strong resilient gel which clears from the mouth uniquely fast. The starch of the preænt 235 invention produces a gel with a unique and distinctive texture compared to .,u,l~. ' starches. The urlique and distinctive texture makes the starch of the preænt invention suitable as a "' for cu.~ iU~ ge]ling gums such as natural gums and gelatin, in whole or in part in food r,.. ~ The starch of the present invention has also been found to produce a more resilient gel than common 2~0 starch. r. ;h~.l,.u.~, it has been found that comstarch produced from maize produces a gel which has improved clarity compared to a gel made from a common starch. Such SUBSTITUTE SHEET (RULE 26 wo95/350t6 2 1 907 6 1 . ~
improved clarity is visible to the human eye and lends itself to a more appetizing foodstuff.
2 4 5 BRIEF DESCRlPI~lON OF THE DRAWINGS
The present invention will now be described, by way of illustration, by the following description and examples with reference to the . , g drawings of which:

FIG I is a graph of enzyme activity for different gene dosages of a single mutant;
FIG 2a is a graph of the DSC scan of waxy, amylose extender and common 255 maize;
FIG 2b is a graph of the DSC scan of a double mutant (~a~t~iAWA WA);
FIG 2c is a graph of the DSC scan of starch from an 2 6 0 (~La ~,AeJ W A W A W A ) .
FlG2d is a graph of the DSC scan of starch from another (wxwxWx/AeAeae).
2 6 5 FIG 3a is a graph of Brabender data of common starch in various pH.
FIG 3b is a graph of Brabender data of waxy starch in various pH.
- FIG 3c is a graph of Brabender data of 70% amylose starch in various pH.

FIG 3d is a graph of Brabender data of double mutant starch in various pH.

SUBSTITUT~ S! IEET (R' 'LE 2 WO 95/3~026 PCT/US9~107828 21 9a76~ -FIG 3e is a graph of Brabender data of a first inter nutant starch in various pH.
2 7 5 FIG 3f is a graph of Brabender data of a second intermutant starch in various pH.
FIG 4a is a schematic showing the design and KStriCtion enzyrne sites of plant ' vectors used to alter gene expression levels of branching 2 8 0 enzyme 1.
FIG 4b is a schematic showing the design and restriction enzyrne sites of plant 1 "" ,_ r .., ~ ;~ ... vectors used to alter gene expression levels of branchingenzyme 11.

FIG 4c is a schematic showing the design and KStriCtion enzyrne sites of plant vectors used to alta gene eXpKsSion levels of bound starch synthase (waxy), 2 9 0 FIG 4d is a schematic showing the design and restriction enzyme sites of plant -- vectors used to alter gene expression levels soluble starch synthase.
FIG 5 is a plot of the elastic modulus (G') over time comparing A gel made 2 9 5 from the starch of the present invention to a gel made from an aewx starch and a gel made from a waxy (~) starch; and FIG 6 is a plot of the elastic modulus (G') plotted against strain for both the pKsent invention and an aewx starch.

SUBSTITUTE SHEET (RULE 26 wo 95/35026 DETAILED DESCRlPI ION OF THE INVENTION
Broadly then the present inYention is an improved crop line which has , ' I e~pression of at least two starch-~y..~ ;"~ enzymes which alter the 3 0 5 amount and type of starch, and, , 1!" alters the grain produced by the plant.
It has been discovered that plants which contain at least two genes which partial1y down regulated or reduce the ætivity of specific isofomms of enzymes in the starch synthesis pathway will ~JI;~;II~,ly produce significant amounts of starch in the 310 grain and will produce altered starch types.
Specialty maize or mutant plants differ from "nommal" maize because of its altered endospemn. The changed endospemm gives rise to a high degree of starch branching, or changed sugar content, or different kemel structure. The endospemm of 15 course is forlned by the spemn and ovule, and the selection of both parents effects the makeup.
The present invention can be formed by two principle methods. The invention can be fommed within a selected crop species by the use of mutant breeding. And the 320 invention can be formed in various plants by the use of ~ of the plants with genes which partially down regulate two or more enzymes in the starch synthesis pathway. More particularly, down regulation of one of the isoforms of the enzyme in the starch synthesis pathway to ,.~ , 1/3 of the nommal ætivity and 2/3 of the normal ætivity in the other isoform enzyme or down regulation of both isoform 325 enzymes in the starch synthesis pathway ~", '~, 2/3 of the nommal activity.
Eæh of these methods has its own ædvantages.
First, the use of mutants to develop unique grain and starches in cereal crops is widely known. However, the present invention is highly unique and surpnsing because Il SUBSTITUTE S~IEET (RULE 26 WO 9~/35026 p~ " c 330 it was expected to prr~duce grain having nortnal starch ~,llola~i~,l;~li-,~. The following table explains how unexpected the present invention iS.
~ .
3 3 5 Genotyloe of Parents Genotyp Type of Sta~h (f~male Ist) of endosplm Starch Ybld Wild-Type MYM AM Normal Nomlal 3 4 0 Gene dosage AAi'aa AAa Nomul Nomlal aa#AA aaA Normal Normal Singlemutant aai'aa aaa Altered Lowered 345 Doublemutant a~Vbb*a~Vbb aaa/bbb Altered Lowered PXESI~NT INV~NTION - EXPECTED RI~SULTS
Intemmut~nt aa/BB~M/bb aaAlBBb Normal Nonnal AAlBBi'LILVbb AALVBBb Nomlal Nomlal 3 5 0 aLvbb~AAlBB aaA/bbB Nomlal Nomlal aalbb~aalBB ar~vbbB Nomlal Normal aalBB~aLvbb aa~vBBb Normal Nomlal PRESENrl.. vr,~ ACrUALRESULrS
355 Int~nnruant aolBB~AAlbb aaA/BAb Altcrc4 Mc41umlo AAalBBb AlJc~cd h;~h - 70%
odbb~AA/BB aaAlbbB A~r~4 of Irornu aolbb~oalBR an~VbbB Alttrc4 3 6 0 aolBO~o~/bb aaalBBb Alt~rc4 na = mut~nt gene " , O
AA = wild tyloe (or non-mutant homoygous gene) i~ = signifies a ~ ; '" ' between two varieties 3 65 arvbb = two mutant genes (both homoygous) SUBSTITUTE SHEET (RULE 26 wo ss/3so26 2 1 9 Q7 6 l r~ L~
Clearly, since the genotype of the endosperm of the present invention shown in the table did not have completely recessive genes, the starch yield and structure was expected to be normal. In fact, according to the present invention, the grain does not 370 evidence normal starch structure altered starch. Historically, when altered starch is produced only small quantities are usually produced. The altered starch of tbe present invention was . ~, produced in larger than expe~ted quantities. Additionally, production of this starch is much simpler than the production of double mutant crops.
Previously only single mutant hybrids have been used extensively for large-scale starch 375 production. Previously, to develop the double mutants, both parents had to carry both mutations which requires significant research and d~ efforts and results in poorstarch yield and poor seed g ~ y. Heretofore, only small-scale production of thedouble mutants has been possible.
3 80 The present invention: . a method of producing gr~un v~ith altered starch qualities which includes the steps of planting a parent which is capable of flowering, this parent having ' "~ complete reduction of at least one specific isoform of an enzyme (A) in the starch synthesis pathway and having no reduction of at least one other specific isoform of an enzyme (13) in the starch synthesis pathway. The 3 8 5 other parent has no reduction of one isofolm of an enzyme (A) and ' "~ complete reduction of at least one other specific iwform of an enzyme (B) in the starch synthesis pathway. It is then necessary to eliminate said first parents capability to produce pollen and allow pollination to proceed from said second mutant parent, and finally harvesting the grain produced by said first parent. Additionally, the method can include the 3 9 0 extracdon of starch from the grain and using said starch as a specialty starch for a variety of uses for which it is shown to be valuable.
In order to prepare a sol in accordance with the present invention, a slurry is prepared which comprises water and an effective amount of starch of the present invention 3 9 5 and the sol subject to a cooking step to form a paste. Generally, cooking entails raising SUBSTITUT~ SHEET (RULE 26) W09513S026 2 ~ 9~ 7 ~ ~ PCTIUS95/07828 the i , ~ of the slurry to above about the ~ i . of the starch and subjecting the starch to enough shear such that the granules rupture and a paste is formed. It is not necessary that all the granules rupture. Preferably, the sol contains the sta~ich of the present invention in the amount of about 1 to about 20% by weight total sol. 00 The sluniy is cooked at a i . of about 90 C and above to provide thickening prior to adding to the foodstuff. Cooking time is about lO minutes. The sol in accordance with the present invention need not be cooked if the starch has already been subjected to a process which makes it cold water swellable. Cooking generally comprises raising the r ' _ of an aqueous slurry of the starch of the present 4 05 invention to the ~ ~ of the starch and subjecting the starch to shear such that the starch granules rupture and form a paste.
A sol or a thickener ~ ;- of the starch of the present invention is added to a foodstuff in a ~,u..v. ' manner in order to provide the benefits of the starch of the 410 present invention to the foodstuff.
In order to prepare the thickened foodstuff, a sol made in accordance with the present invention is combined with a foodstuff and the . is cooked to tbe necessary degree to provide a thickened foodstuff. Conventional mixing is employed to 415 combine the sol with the foodstuff. Cooking of the sol and foodstuff ~ . is also carried out in a ,u.l ~ ' manner, Altematively, starch of the present invention is mixed ~vith the foodstuff or a slurry comprising the starch of the preænt invention and water is mixed with a foodstuff 42 0 and the resulting mixture is cooked to the desired degree to obtain a thickened foodstuff, When the starch itself or a slurry containing the starch itself is mixed with a foodstuff, the resulting mixture must be cooked in order to provide a thickened foodstuff. The mixing as well as the cooking is r , " ' ' in a ~u~ t;u~ l manner. Cooking is carried out at a: , of about 90 C and above. Cooking time is about lO minutes but may SUBSTITUTE SHEET (RU~ 2~) WO 95~35026 2 1 9 0 7 6 1 PCTIUS9510~828 42 5 vary depending on the amount of foodstuffpresent and the amount of shear that the mix is subject to during cooking.
Such a thickencr c . ~ ;- can provide ' ' ' economic advantage to the user. Those familiar with the art have long uscd a variety of gelling gums for their clean 43 0 breaking texture. Application of the present invention have included but are not limited to gum candies, gellcd desserts, glazes and spreads and can be used to replace ~.u..~. ' gelling gums such as kappa ~ O agar, pectin, or gelatin. These ~.u...~...;~...d gelling gums can be quite expensive however, and have other ~' " . ,, including the presence of off-flavors, lack of hcat or acid stability, limited availability, or lack of Kosher 435 approval. It has been found that the starch of the present invention can replace dl or a portion of these cu~ ' gelling gums.
In order to replace a gdling gum in food r~ . a weight ratio of about 1:1, starch of the present i..~, ' ,. gum, can be employed. Larger or smdler amounts44 O of the starch of the present invention may be used to replace a gelling Oum. Such gelling gums include gelatin, pectin" " gum arabic, tragacanth, guar, locust bean, zanthan, agar, algin and ~I,u~ ' jl cellulose.
Naturally, the starch of the present invention can be qsed in any food ' 44 5 where there is a need to provide gel ~ ` ;`1 ;' ` and a clean brcak from the mouth. For example, the starch of the present invention can be used in a food r ~ '- which had heretofore employed a common starch, thereby providing the food with improved properties, i.e. clean break when compared to the same food ' ' using a common starch.

The clean break of a gel made with the starch of the present invention is useful in a - variety of food nl~ The clean break of the starch gel has vdue in a variety of bakery ~ ;,. " for example cream or fruit fillings for pies such as lemon, banana cream or Bavarian cream; and in low or reduced fat high solids fruit centers for cookies, SUBSTITUTE Sl IEET ~RULE ~6 WO 95/35026 2 1 ~ ~ 7 6 1 PCT/US95/07828 455 for example, in fig bars. The starch of the present invention dso creates an improved Kxture in mousses, egg custards, flans and aspics.
The Krm starch as used in the ~ r; A~ and claims means not only the ,uI,A~,lidl~ pure starch granules as extracted from a starch bearing plant but also grain 4 6 0 products of the starch granule such as flour, grit, hominy and meal.
The following examples of the present invention are given for illustrative purposes only. These examples are not inKnded to limit the type or the uses of the present invention. The present invention or the gr~un or starch or sugar thereof can oe useful in, 465 but may not be limited to, the preparation of foodstuff, paper, plastics, adhesives, patnts, production of ethanol and corn syrup products.
Example I
4 7 0 The physical properties of various e.. ~ " of the present invention (similar genotypes are from different maize crosses). These tables display data well known to people skilled in the arts of evaluating new and novel starches. Data on moistures, and percent oil, protein, solubles and starch are 475 useful in evaluating yield and milling poKntial. Starch DSC
(Differentid Scanning (~ ) data are vduable for evaluating starch cooking and g. 1 ~; A' ;~ . properties. Starch particle size data are vduable for making decisions on starch milling and separation properties. Brabender and starch paste 4 8 0 data are most important for evduating a new starch's potentiaA
for improved food ~ r ~ where particular starch thickening and pasting and i' properLies are mostly desired. Such data when interpreted as a whole collection of ~ enable one skilled in the art to SUBSTITUT~ SHEET (RULE 26 W095135026 2 19 0 7 6 1 r~l~u~ 6 485 decide whether to conduct further more detailed tests of the starches properties and potent;al.
Genotype %Moisture %Starch %Protein %Oil %Solubles aeae/wx 8.01 72.12 11.30 3.63 6.51 aeæ/wx 8.00 7134 11.74 3.79 6AI
aeæ/wx 8.03 67.52 12.6] 3.67 7.01 aeae/æ 8.13 60.11 14.59 6.60 10.14 sulsul/wx 8.18 69.66 12.09 4.03 6.91 dudu/wx 7.98 71.10 11.57 3.92 6.61 dudu/sul 7.90 70.53 11.67 3.93 6.81 flfl/o 7.78 70.23 1 1.60 4.23 6.59 sulsul/du 7.81 69.00 12.20 4.M 7.98 sulsul/æ 7.88 69.03 12.27 4.88 8.02 sulsul/su2 795 69.67 12.28 4.55 7.49 .
.

SUBSTITUTE SHEET (RULE 26) WO 95/3s026 . 2 1 9 0 7 6 ~ PCT/US95/07828 ~9 0 Table 2 - Corn Data S93 % % % % %
EDIr,~ l~n~ Row Moisture ~ta~h l~ill ~lil ~QIIlk~
] aeaewx 4660 9.15 64.80 14.58 4.47 7.64 2 aeaesu 4636 9.27 66.25 14.77 4.54 7.40 3 aeaedu 4612 9.24 66.01 IA65 4.73 7.80 4 wxwxae 4600 9.07 65.72 ]3.91 A38 8.]4 5 wxwxsu 4648 9.06 66.03 ]4.79 4.08 7.83 6 wxwxdu 4624 8.77 65.32 ]5.62 427 8.23 7 susuae 4594 8.85 64.83 13.11 4.71 8.49 8 susudu 4672 8.73 65.39 ]3.20 5.01 8.35 9 susudu 4618 8.77 65.]5 14.52 435 8.26 ]0 sing]eae 4582 9.]4 6052 ]5.]9 5.95 9.29 ] ]single wx 4654 8.83 65.44 ]4.04 4.77 9.43 12 single su 4630 7.75 54.98 14.25 6.50 9.78 13 duduae 4588 8.97 6926 1230 A89 6.85 ]4 duduwx 4666 8.88 70A3 ] 1.25 4.40 6.96 15 dudusu 4642 9A8 70.43 13.48 4.67 6.92 16 single du 4606 850 65.54 12.02 6.03 933 - Starch Data S93 % % % % %
Entr.~ ~2~1YQ ~ ~Q~ ~ ~ ~ ~ ~8 aeaewx 4660 3.82 86.44 1.68 0.18 31.60 597.1 2 aeaesu 4636 6.51 86.95 1.56 0.20 31.74 603.9 3 aeaedu 4612 5.22 86.44 1.70 0.11 3338 598.8 4 wxwxae 4600 733 87.89 0.39 0.09 21.10 601.1 s wxwxsu 4648 7.27 85.87 1.32 0.09 2A.68 591.7 6 wxwxdu 462A 7.31 86.07 1.12 Q10 24.57 595.2 7 susuae 4594 8.19 86.32 Q93 0.07 29.C0 604.5 8 susudu 4672 8.09 87.16 052 0.08 27.86 599.6 9 susudu 4618 7.67 84.20 0.60 0.12 3Q82 ~501.1 10 sing]eae 4582 7.16 8331 1.37 0.15 6555 6Q~I
Il single wx 4654 8.30 85.16 0.51 0.14 0.43 xx 12 single su 4630 6.82 80.92 5.08 xx 29.32 598A
13 duduae 4588 6.58 84.31 0.62 0.17 29.86 xx 14 duduwx 4666 11.08 84.77 0.48 0.16 28.74 ~x 15 dudusu 4642 8.48 87.34 Q62 Q16 26.07 xx 16 singledu 4606 8.36 8Q70 0.46 0.08 37.88 608.9 SUBSrlTUT~ SHEET (RULE 26) 4 9 5 Table 2 - (Starch Data) %qO % %
aeae/wx 9.56 83.84 0.63 QQS 29.44 593.1 2 ae ae / wx 7.96 85.94 0.70 0.05 27.89 596.6 3 ae ae / wx 11.71 82 54 0.82 Q05 42.54 595.1 4 aeae/ae 6.Q9 84.17 12A 0.14 63.60 602A
5 sul sul /wx 4.37 90.00 Q56 0.Q9 28.36 600.0 6 dudu/wx 5.95 8850 0.61 0.Q9 27.98 596.9 7 dudu/sul 6.93 82~66 Q69 0.10 29.98 600.8 8 flfl/0 9.24 8272 093 0.14 2929 596.8 9 sul sul /du 9.01 84.82 Q71 Q02 2957 600.8 10sul sul /ae 7.78 84.81 0.63 0.03 29.90 6C0.8 Ilsul sul /su2 7.61 85.02 0.49 QQ2 29.68 6023 12White waxy 1834 9A94db 0.77 Q09 2.74 526.9 - Starch DSC Data Pellk Delta H Peek n Onset Endset sL cc l¢g C C C
aeae/wx 67.7 11.67 IQ0.0 62.6 76.6 2 aeae/wx 665 10.33 96.8 61.9 74.8 3 aeae/wx 7A6 IQ00 100.8 68.8 83A
4 aeae/ae 81.0 11.83 xx 66.0 107A
5 sul sul /wx 69.7 1200 98.3 64.2 77.7 6 dudu/wx 72.2 1150 99.5 66.6 82.3 7 dudu/sul 702 9.67 98A 65.7 78.3 8 fl fl/0 71.4 12.33 100.9 67.1 8Q7 9 sul sul /du 68.8 7.83 995 63.2 79.1 10sul sul /ae 68.1 IQ33 99.8 61.9 78.3 Ilsulsul/su2 67A 12.00 96.1 61.3 79.5 12White waxy 72.7 1533 xx 66.1 82.6 - Starch DSC Data S93 Pe~k Delta H Peek n Onset E.ndset ~lY~a~k~ ~ oc J/~ C C ~
Deaewx 4660 73.8 11.17 99A 67.7 82.8 2 aeaewx 4636 73.0 11.50 101.0 66.6 83A
3 aeaewx 4612 73.3 10.83 98.6 66.7 83.0 4 wxwsae 46~0 72.9 12.83 99.9 67A 81.7 5 wxwssu 4648 73.7 12.17 98.2 682 81.6 6 wxwsdu 4624 73.6 1133 99.6 69.3 80.5 7 susllae 4594 72.0 IQ33 97.7 67.2 79.7 8 susudu 4672 71.9 9.83 98.9 67.0 79.1 9 susudu 4618 725 1033 97.0 68A 79.8 10single ae 4582 84.6 15.67 xx 68.7 106.9 I lsingle wx 4654 72.5 16.17 ~ 68.1 81.4 12single su 4630 709 15.50 100.8 63.6 772 13duduae 4588 71.8 1350 1005 67.1 81.1 14duduwx 4666 71.9 10.17 98.8 67.2 79.5 15dudusu 4642 71.7 IQ17 97.2 66.8 79.0 16singledu 4606 71.6 9.50 100.1 65.8 79.8 .
SUBSTITUTE SHEET (RUL~ 26~

W0 95/35026 ~ &
2 ~ 9376 1 Table 2 5 0 0 - Brabender Data S93 lR HP HF CP CF
ntrY73ackw~und ]35~ ~BI~ C L3U . 1~_ }3U ~
aeaewx 4660 460g/5.5% 65.0 315 285 940 940 2 aeaewx 4636 460g/5.5% 81.5 225 225 470 430 3 aeaewx 4612 460g/5.5% 84.5 180 180 410 380 4 wxwsae 4600 460g/5.5% 74.0 360 355 590 460 5 wxwssu 4648 460g/55% 74D 325 325 540 470 6 wxwsdu 4624 460g/5.5% 80.0 315 270 480 425 7 susuæ 4594 460g/5.5% 77.0 270 270 610 585 8 susudu 4672 460g/5.5% 77.0 295 295 785 610 9 susudu 4618 460g/5.5% 77.0 295 295 620 620 10 single ae 4582 460g/12% 90.5 170 170 240 240 I l single wx 4654 460g/5.5% 68.0 750 360 415 405 12 single su 4630 90g/55% 93.5 35 35 40 40 13 duduae 4588 460g/55% 83.0 225 225 545 520 14 duduwx 4666 460gl5.5% 86.0 245 245 600 550 15 dudusu 4642 460g/55% 84.5 270 270 620 585 16 single du 4606 460gl5.5% 89.0 70 70 160 160 IR. = n~it~ isc CP = cooling pe~k CF = cooling final - Brabender Data IRHP HF CP CF
C~ ~ ~ ~
ae æ / wx 460g/5.5% 83.0 220 220 520 460 2 .aeae/wx 460g/5.5% 89.0 220 220 540 510 3 aeae/wx 460g/5.5% 845 270 270 510 450 4 aeae/ae 460g/129~o 90.5 490 490 1220 845 5 sul sul /wx 460g/5.5% 80.0 250 240 595 535 6 du du / wx 460glS5% 83.0 280 250 560 470 7 dudu/sul 460g/5.5% 50.0/83.0 230 230 575 535 8 flfl/0 460g/5.5% 81.5 280 255 630 560 9 sul sul /du 460g/5.5% 84.5 200 200 500 460 10sul sul /ae 460g/5.5% 86.0 205 205 455 415 Ilsul sul /su2 460g/5.5% 84.5 180 180 420 385 12White waxy 90g/5.5% 68.0 830 220 310 270 SUBSTITUTE SHEET (RULE 26) woss/3so26 2 ~ ~ 0 7 6 i 1~l,., ",~
Table 2 - Starch Particle Size Data (Volume Distribution) S93 % Starch ~Y Backeround Row Mode ~n Mean ~un ~di3nJL_ Re~over,v aeaewx 4660 16.90 12.57 16.04 45.9 2 aeaewx 4636 16.63 11.82 15.23 57.9 3 aeaewx 4612 16.36 11.48 15.20 52A
4 wxwsae 4600 17.43 11.62 16.53 41.5 5 wxwssu 4648 17.72 12A3 16.59 58.8 6 wxwsdu 462A 17.48 12.22 16.26 54.1 7 susuae 4594 16.36 11.96 15.66 55.5 8 susudu 4672 16.65 9.83 15.29 58.7 9 susudu 4618 16.36 11.82 15.61 55.9 10 single ae 4582 13.09 9.07 12.25 64.0 Il singlewx 4654 18.03 11.99 16.85 55.7 12 single su 4630 7.43 5.16 7.95 5.6 13 duduae 4588 16.90 11.01 15.37 SQ9 14 duduwx 4666 17.19 11.85 16.21 52.5 15 dudusu 4642 17.17 12.22 16.01 44.9 16 singledu 4606 14.41 9.77 13.19 58.9 - Starch Particle Size Data (Volume 1~ ' ) 9~o Starch E~~Ll;L ModeILm ~LU_ ~ Recoverv aeae/wx 16.63 10.90 15.70 51A
2aeae/wx 16.12 10.44 14.92 52.8 3ae ae / wx 15.60 10.55 14.60 59A
4aeae/ae 11.53 8.40 1130 63.7 5sul sul / wx 14.88 IQ2A 14.21 52.5 6du du / wx 16.38 IQ73 15.12 52.0 7dudu/sul 15.87 11.03 14.98 53.2 8flfl/0 16.12 10.73 1535 69.3 9sul sul /du 15.87 10.73 14.92 53.7 10sul sul /ae 15.11 7.85 14.35 48.5 I lsul sul / su2 15.85 IQ33 14.20 58A
y17.1 845 1~81 71.5 SUBSTITUTE: SHEET (RULE 26~

Wo 95135026 2 1 9 0 7 6 1 PCT~S9S/07828 515 Table 2 - Starch Paste Data BrQokfield Freeze Viscosity Gel-2Ahr Thaw EDg:Y Genot~e CPS. 20 rvm ~m~ cles ~eæ/wx 6,200 166.6 0 2 ae ae / wx 6,700 2272 0 3 æ æ / wx 8,000 256.3 0 4 ææ/ae 14,250 111.1 0 5 sulsul/wx 6,800 173A Q
6 dudu/wx 6,000 141.4 0 7 dudu/sul 7,100 215.4 0 8 fl fl / 0 7,500 209A 0 9 8ul sul /du 6,300 219.7 0 10sul sul /æ 6,000 172.2 0 Ilsul sul /su2 5,500 158.8 0 12White waxy 1.900 15.3 3 - Starch Paste Data Brookfield Freeze S93 Viscosity Thaw ~EYGenQtvee Row = ~P,C 2n Q~
aeaelwx 4660 25,000 151.8 0 2 aeæ/wx 4636 6,500 240.1 0 3 ææ/wx 4612 3,150 216.6 0 4 ae æ / ae 4600 5,600 159.5 0 5 sul sul /wx 4648 6,100 126A 0 6 du du / wx 4624 5,700 114.4 0 7 du du / sul 4594 8,700 254.8 0 8 fl fl /0 46?2 12,000 272.8 0 9 sul sul /du 4618 10,250 233.6 0 10 sul sul /æ 4582 2,500 53.7* 0 I Isul sul / su2 4654 3,100 13.3 0 12 White WIXy 4630 275 29.0 3 13 duduæ 4588 7,200 226.5 0 14 duduwx 4666 7,300 233A 0 15 dudusu 4642 5,100 3329 0 16 single du 4606 1,650 99.3 0 $ came out as a plug SUBSI ~TUTE SHEET (RULE 26 w095/35026 2 1 9076 1 ~."~ ",~"
DEFlNlTIONS:
DIFFERENTIAL SCANNING CALORIMETRY (DSC) IR denotes initial rise 525 HP denotes heat peak HF denotes heat final CP denotes cooling peak CF denotes cooling final.

The Brookfield Viscometer measures shear-strength (in centipoise, cP) and stability of starch pastes.
BRABENDER VISCO-AMYLOGRAPH DATA
53 5 Pasting i , denotes the i , of paste formation.
Peak Viscosity denotes the . . _ needed to provide a useable paste.
Viscosity at 95C denotes the ease of cooking of the starch.
Viscosity at 50C denotes the setback in paste viscosity during cooling of a hot paste.
540 Viscosity after I hour at 50C denotes the stability of the cooked paste.
ORN PERCENT PROTEIN, STARCH, OIL AND MOISTURE
of oil starch and protein m com give a measure of starch yield how ,~, . ~,.~1~ the starch is.

STARCH PERCENT PROTE~, STARCH, OL AND MOISTURE
re. ~ of oil starch and protein in starch give a measure of how well purified the starch is and indicates millability.

These data provide a measure of apparent amylose levels in starch.

SUBSTITUTE SHEET (RULE ~6 woss/35026 21 ~76 1 STARCH PARTICLE SlZE DATA
Starch particle size gives an indication of starch yield and ~u._.~;lily through5 5 5 the milling process.
SHORT HAND
aeaewx in table 2 refers to aeaeAE/wxWxWx, likewise 1' ' A_' ' r)U/WXWXWX Throughout this table the wild type is not listed.
Figure 1 is a graph of enzyme activities for individual gene-dosages (e.g., MMM,560 ~MM, mmM, mmm) of mutant alleles of the single mutants of amylose extender and dull. These data show the enzyme activities of sucrose synthase (SS), UDP-glucose ~ yl~x, (UDM-PP), O' ' (GK), rl u~luki,.~., (FK), (PGM), ~ , O isomerase (MI), ATP-dependent 1' , ' ' u~lvl~ill~ (PFK), PPi dependent I ' , ' ', ' (PFP), ADP-565 glucose ,u,,' yl~_ (ADM-PP), soluble starch synthase (SSS), branching enzyme (BE) and bound starch synthase (BSS). Enzyme activities are presented as relative to the wild-type control (MMM). In the case of the full mutants (mmm) there is a dramatic effect on expression levels of various enzymes in the pathway of starch synthesis. In the case of the partial mutants (mMM and mmM) 5 7 0 there is almost no detectable change in expression levels. These data mdicated that the alteration in starch quality observed with the single mutants is a . of the u . _.~ A,U.~;VII of æveral enzymes as well as elimination of the enzyme coded for by the mutated allele. By combining two mutant doses (e.g., wxwxWx) with other doses of another mutation (e.g., AeAeae) there would be partial reduction in two575 enzymes without the U._~AIJ' seen in the rest of the pathway.
hgure 2 is a graph of the DSC scan of starches extracted from grain taken frorn waxy, amylose extender and common (wild type) com. Such DSC scans enable one skilled in the art to provide numerical data (see tables in text for data on Pea~A
Tc r ', Delta H, Peak II T~ . , Onset Temperature and Endset 5 8 0 Temperature). It is particularly noteworthy that the profile of the high amylose starch is different from the common starch and waxy starch.

SUBSTITUTE S~IEET (RULE 26 WO 95~026 2 1 9 0 7 6 1 PCTIUS95107828 Figure 2b is a graph of the DSC scan of starches extracted from grain taken fromthe double mutant (aeaeae/wxwxwx) com. Such DSC scans enable one skilled in the art to provide numerical data (see tables in text for data on Peak T; . Delta 585 H, Peak n T~ . , Onset Temperature and Endset Temperature). It is particularly noteworthy that the profile of the double mutants is different from the data provided in Figure 2a on common starch and the single mutants, waxy and high amylose.
59 0 Figure 2c is a graph of the DSC scan of starches extracted from grain taken from (aeaeAe/WxWxwx) com. Such DSC scans enable one skilled in the art to provide numerical data (see tables in text for data on Peak T , Delta H, Peak ~ T~ r ' , Onset Temperature and Endset Temperature). It is particularly noteworthy that the profile of the intermutant starch is different from the starch of the 595 double mutant and appears to be similar to that of waxy starch.
hgure 2d is a graph of the DSC scan of starches extracted from grain taken from (wxwxWx/AeAeae) corn. Such DSC scans enable one skilled in the art to provide numerical data (see tables in text for data on Peak T~ , ` . Delta H,600 Peak 11 Temperature, Onset Temperature and Endset T: , ). It is particularly noteworthy that the profile of the intermutant starch is different from the starch of the double mutant and appears to be similar to that of waxy starch.
Figure 3a is a graph of Brabender data taken from common starch in either neutral 605 or acid conditions. Common corn starch shows substantial breakdown in viscosity using acid conditions.
F~gure 3b is a graph of Brabender data taken from waxy starch in either neutral or acid conditions. The waxy mutation most particularly affects viscosity of the starch 610 in neutral conditions.

SUBSTITUTE SHEET (PULE 26 WO 95/35026 . 2 ~ 9 ~ 7 ~ 1 PCT/US95/07828 Figure 3c is a graph of Brabender data taken from amylose extender (70%
amylose) starch in either neutral or acid conditions. High amylose starches increase in viscosity in either acid or neutral conditions, Figure 3d is a graph of Brabender data taken from double mutant WAWA) s~arch in neutral conditions. Double mutant starches maintain Yiscosity despite being IIVIIIV~J~.JU;~ for the waxy mutation.
62 0 Figure 3e is a graph of Brabender data taken from intemmutant (aeaeAe/WxWxwx) starch in neutral conditions. It is particularly noteworthy from these data that the new intermutant starches provide an increasing strength of viscosity similar to tbat seen with high amylose mutants, despite containing no increase in apparent amylose content.

Flgure 3f is a graph of Brabender data taken from intermutant (wxwxWx/AeAeae) starch in neutral conditions. It is particularly noteworthy frvm these data that the new intermutant starches providc an increasing strength of viscosity similar to that seen with high amylose mutants, despite containing no increase in apparent amylose 630 content.
Example 2 This example illustr~tes the production of maize grain 635 possessing starch of the present invention. Maize plants of various 1~.. ' can be converted to mutant genotypes using either traditional breeding and/or v~h.lua~
techniques or else using ,, such as chemical treatments of pollen. ~ the altemative, waxy mbreds and 6~ 0 hybrids can also be purchased from a number of suppliers and foundation seed companies. Any maize line with good SUBST~TUTE ~HEET (RULE 26) WO 95/35026 PCrlUS95107828 2191~
agronomic traits and relatively high yield can be employed. In the present invention, normal inbred lines are converted to mutant inbred lines using chemical ,, followed by 6 4 5 careful selection of the mutant grain type from the segregating offspring. This method is well known to those skilled m the art (see for example, Neuffer, M.G. and Chang, M.T. 1989.
Induced mutations in biological and agronomic research.
Vortr. I`f' .g 16, 165-178). Any '~, 650 valuable inbred line may be used for this process. The lines were confirmed to carry the mutation of interest by an allelism test in which the line may be crossed with a known mutant line, a process well known to those skilled in the art.
r. i ~, the kernels from the line will have the 655 appearance and iodine-staining ~ typical of the mutation selected, a method well known to those skilled in the art. In order to obtain the highest yields from the plants it is best next to produce a hybrid cross between two inbreds, both inbreds carrying the same mutation (e.g., both inbreds being 660 waxy or amylose extender types). It is preferred to produce two hybrids, one being the male and being l.~ L~b~.,D for one mutation and the other being the female and being IIU~ IL~6~JUD for the other mutation. The male and female hybrids can be made up of the same or different genetic 665 1 ~ ' it is merely import3nt for the two lines to have similar maturities in the field (i.e., require similar heat units from, to silking and pollen shed). In order to make the intermutant aoss in the field it is necessary to eliminate pollen production from the female. This can h 670 done by a variety of methods including, but not limited to, hand pollination, hand and mechanical ~'~

SUBSTITUTE SHEET (RULE 26) WO 95/35026 2 ~ 9 0 7 6 ~ PCT/US95107828 v6.~ genetic or ~: r~ male sterility into the female plants, introducing male sterility through genetic r ~ ti.~ll and use of chemical detasseling agents. The 675 grain of this crvss contains the present invention with a genotype in the endosperm of aaAlBBb, with the starch froln this genotype called intemmutant starch. It is well known to those skilled in the art that the genetic v~k6.. ' can be optimized for best starch qualities.

Example 3 Starch may be extracted from gr~un by a number of different methods. The most commonly used method involves a "wet 685 milling" procedure known and used throughout the world The basic principle involves steeping and starch separation.
The key step in this process involves softening the grain in a steep tank a process which has been optimized to permit optimal separation of the com grain ----r This method 690 was employed to extract the starch, wxwxWx/AeAeae from the grain of an intermutant developed in accordance with Example 2. The gemm was easily liberated intact and freed from adhering endosperm and hull. The endospemm is macerated under water, the starch was easi]y separated as a 695 white floc and gluten proteins are obtained as a yellow floc.
The grain was steeped for 30-40 hrs at 48-52C in tanks usually holding 50-90 metric tons of grain. The steep water contains 0.2% sulphur dioxide (SO2 gas is bubbled-in) and so is mildly acidic (pH 4.0). The sulphur dioxide helps break-up 700 the protein matrix permitting the endosperm matrix to break-up into granules. After steeping, the grain were coarsely SUbSTlTUTE SHEET (RULE 26 w0 9~/35026 2 1 9 0 7 6 ~ 8 ground or pulped. The oil-rich embryo floats Io the surface and dense starchy endosperm sinks, Separation is achieved through hydreclones (continuous separation). The starch was 705 purified further after being milled through "an impact mill known as Entoleter mill" which smashes the slurry at high speed through counter-rotating grooYed plates made of hardened steel alloy, followed by impact with an outer impact ring. The defibered starch was separated from gluten by 710 ~ .~;r.. ~,. ;.. to give two fractions: protein (70% protein) arld starch (2% protein). Processing i , ~ was maintained above 45C to prevent microbial growth. The starch was dried by flash-drying by injection into an air stream heated to 200 260F.

Varjous intermutant grains can have tbe starch purified and prepared in this manner is suitable for a variety of food, feed and industrial uses. It may be used directly as unmodified com starch. It may be modified by chemical or physical treatments 72 0 that preserve granule structure and granules may be washed to remove residual reactants. Bleaching is sometimes used lo create , ..' starches. The starch ean be gelatinized using a high l . treatment and sold directly as gel$inized starch. Such starch may be ehemieally modified 7 2 5 and dried. The polymer itself may be hydrolysed partially or completely to produce ' ' or glucose. Such products ean be further modified by F~ to produce ethanol for the gasoline industry, or the glueose ean be converted to high-fructose corn syrup for the sweetener 7 3 0 industry.

SUBSTITUTE SHEET (RULE 26 w0 95135026 r~l,u~
2 1 9~76 ~ --Example 4 The mutations called shrunken-2 (sh2), brittle-2 (bt2), dull 735 (du), sugary (su), waxy (wx) and amylose extender (ae) encode isoforms of ADP glucose ~,~ U ~JIIUI ~l~, enzyme, soluble starch synthase, bound starch synthase and branching enzymes:
740 Shrunken-2 encodes one subunit of ADP glucose Brittle-2 encodes one subunit of ADP glucose ~v~ h~
Waxy encodes granule bound starch synthase, 745 Amylose Extender encodes an isoform of branching enzyme, Dull alters expression of an isoforms of soluble starch synthase and branching enzyme, Sugary alters expression and activities of soluble starch synthases and ' ' ' ~ enzyme.

Using known mutants and the gen~dosage crossing regimes we have examined the effects of altered gene expression on starch deposition in griun (See figures). With the bt2 mutan~ we see a progressive loss in measurable ADP
755 Glc l~J.. ' ,' ~' activity which correlates well v~ith a loss in starch synthesis in the grain. The control strength exerted by this enzyme over flux to starch cannot be quantified from these data. In fæt our studies indicate that this enzyme is one of the major 1' of the duration of 7 60 starch synthesis and may have little cont~ol over rate of starch synthesis. This mutation does not appreciably alter starch SUBSTITUTE SHEET ~RULE 26 w095135026 2190761 r~ s structure. When the mutations are with sugary. dull, waxy and amylose extender we now do detect changes in starch fine structure (branched chain length changes as well as changes in - 765 ~.. ,~ i.. ratios). In these cases there is more minor control of flux to starch (except with the sugary mutant which is used to make sweet-com genotypes). In all of these cases it is the changes in ratios of the starch synthases and branching enzymes which have resulted in alterations in starch 770 fine structure. A dramatic new finding in these studies was the discovery that not only does the mutation reduce expression of key enzymes, but also it induces an u. .~ - - of other enzymes in the pathway. r. IS, it is only in the full mutant (mmm) genotypes where we see changes in starch fine 775 structure .~ that the structural changes occur only when there is an enzyme isoform loss in u ' - with an enzyme isoform u. , Whilst not wishing to be bound by this proposal, these data illustrate the means by which starch structure may be influenced by not only reducing 7 8 0 expression (eg using antisense constructs) an enzyme but also be ' '~ increasing expression (eg using sense construct).
Plant ~ r, vectors for use in the method of the invention may be 785 constructed using standard techniques. Since these enzymes are localized in the amyloplast . , of the cell, the gene construct requires the presence of an amyloplast transit peptide to ensure its correct localization in the amyloplast. The construct may carry the gene either in the partial ænse orientation or in the antisense orientation. Expression of said gene in the plant results in a reduction 790 in expression of the enzyme by effects well known in the art as "sense . .,or antisense". When only a reduction in expression is needed the transit peptide is not SUBSTITUT~ SHEET ~ULE 26) wo 9~J3~026 9 ~ 7 6 1 ~ liD
required. However, when enzyme U.vlvhlJlCDD;vll iS rvequired then a correct piastid targeting se~iuence is needed in the construct. Key enzymes required for this invention include branching enzyme and soluble and bound starch synthase.
795 Branching enzyme [1,4-a-D-glucan: 1,4~-D-glucan v~ -D-(1,4-a-D-glucano ) transferase] convertD arnylose to .u.-.t' r ", (a segment of a 1,4-a-D-glucan chain is transferred to a primary hydroxyl group in a similar glucan chain) sometimes cailed Q-enzyme. Soluble starch synthase [ADPglucose: I ,4-a-D-glucan 4-a-D-vUD~ ;lal~D~v~aDC] extends the ~ of alll~5v~vlill and perhaps also 8 0 0 amylose. Bound starch synthase LAi~Pgiucose: I ,4-v~-D-glucan 4-a-D-yltl~lDfvlaDv] extends the chain length of amylose and perhaps aiso alll,r~utfvvlill~
For any antisenæ or senæ- . . construct only a partiai cDNA clone is 805 re~iuired to be expresæd in a transgenic plant. Where enzyme U~v~CALllvDD;UII iS
rc~iuired, then a full length cDNA clone is needed. The se~iuence of maize branching enzyme-I waD ~ , ' by Baba, T., Nishihara, M., Mizuno, K., Kawasai~i, T., Shimada, H., KobayaDhi, E., Ohnishi, S., TanalAa, K, and Arai, Y. (1~1. . ;r;.~
cDNA Cloning, and Gene ~xpression of Soluble Starch Synthaæ in Rice (Oryza-810 sativaL) Immature Seeds. Plant Physiology. 103:565-573, 1993) . Starch branching enzyme-ll from maize endosperrn was ~ ' by Fisher, D.K., Boyer, CD., and Hannah, L.C. (Starch Branching Bnzyme-ll from Maize Fn~1n~i~rm Plant Physiology. 1021045-lvn46~ 1993). The article by Mu, C., Harn, C., Ko, Y., Singletary, G. W., Keeling, P. i,. and Wasserman, B. P. shows an association of a 815 76i~Da l,ul.~ livie with soluble starch synthase I ætivity in maize (cv B73) endosperm. Plant Joumai 6, 151-159 (1994). The maize waxy locus for UDP-glucose starch glycosyl transferaæ was cloned in 1986 by Kioesgen, R.B., Gierl, A., Schwarz-Sommer, Z. and Saedler, H (Molecuiar anaiysis of the waxy locus of Zea. Mol. Gen. Genet. 203, 237-244). Recently, the sequence for the maize sugary 820 locus was ûbserved by James, M. and Wright, A. (The Plant Journai) using transposon ~, to locate the gene. The gene for any such i~rotein is thought SUBSTITUTE SHEET (~ULE 26 -WO 95/35026 2 l 9 0 7 6 1 PCTIUS95107828 to be a 1' ' ' ~ enzyme and may be used in constructs according to this invention.
82 5 It is believed that the chloroplast transit peptides have similar sequences (Heijne et al describe a database of chloroplast transit peptides in 1991, Plant Mol Biol Reporter, 9(2)~ 126). Other potential transit peptides are those of ADPG
(1991, Plant Mol Biol Reporter, 9:1û4 126), small subunit RUBISCO, ' synthase, ~1~ '' ' /Je-3P-d~.b~ub~ c and nitrite 830 reductase. For examp~e, the consensus sequence of the transit peptide of small subunit RUBISCO from many genotypes has the sequenoe:
MASSMLSSAAVATRTNPAQAS~ VAPFTGLKSAAFPVSRKQNLDlTSlAS
NGGRVOC
The com small subunit transit peptide of RUBISCO has the sequence:
8 3 5 MAPI`VMMASSATATRTNPAQAS AVAPFQGLKSTASLPVARR.~R.~T r.
VASNGGRIRC.
The transit peptide of leaf starch synthase from com has the sequence:
MAALATSQLVATRAGLGVPDAS TFRRGAAQGLRGARASAAADTL
SMRTASARAAPRHQQQARRGGR FPSLWC.

Example ~
Production of fertile transgenic maize plants has been done since 1990. Although a number of DNA deliwry systems are 845 known, the selection is a particle ~ ' .' As noted above, constructs of the various maize mutant genes are available from ~ in the U.S. and Europe. Attached are a few examples of some of these constructs as shown in Figures 4a - d. Figure 4c shows a promoter, which is CaMv 850 (cauliflower mosaic virus), an Adhl, the waxy gene, the and the pat gene which is useful as a selectable SUBSTITUTE SHEET (RULE 26 wo g~ 026 2 1 9 0 7 6 I PCT/[IS95/078Z8 marker and amp. Figure 4d is similar but shows the soluble starch syntheses first isoform gene in the construct. Figure 4a again has the same construct but shows the branching enzyme 855 first isoform. Figure 4b shows the second branching enzyme second isoform. Of course, other constructs associated with the gene mutants used in maize breeding are also available.
For purposes of this example reference is made to Figure 4c, the waxy construct.8 6 0 The purpose of this experiment is to form an inbred that has partial down regulation of the waxy gene. If the inbred selected is already a mutant for ae, then the gr~un produced by crossing with a non-mutant inbred will be the grain of an Depending on the strength of the down regulation, the female inbreds grain will resemble the mm*tmmY' or the mm*tm~ type of starch and grain. Clearly, the 865 ~.~, r~ " allows a more precise way of down regulation of the starch synthesis activity such that the alteration of tbe starch can be finely tuned.
To assure reasonable levels of down regulation of the waxy gene, the target tissue is immature zygotic embryos, through cll~blylO
870 callus can also be employed. Immature zygotic embryos from A188 plants 12 days after pollinated with the B73 ae inbred can be selected. The medium for the callus was 6 mM L,proline, 2% (w/v) sucrose, 2 mg/l 2,4- d;~ lu~ acid (2,4-D) and 0.3% (w/v) Gelrite (Caroline Biological Supply) (pH 6.0). Callus is grownand suspension cultures were initiated.

A MS-based liquid medium containing 100 mg/l myo-inositol, 2 mg/l 2,4-D, 2 mg/l l-l .' ' ' ~ acid (NAA), 6 mM proline, 200 mg/l casein hydrolysate (Difco T ' ), 3% (w/v) sucrose, and 5% (v/v) coconut water (Difco T ~ ) (pH 6.0). Cell , were maintained in this medium in 125 ml 880 r ~ , . flasks at 28C in the dark on a gyrating shaker at 125 rpm.

SUBSTITUT~ SHEET (RULE 2~) wo ssl3so26 2 1 9 0 7 6 ~
The . ' Il.dli~". vector in Figure 4c is selected. This plasmid contains a 355-ladh-pat nos 3' selectable gene expression cassette.
8 85 The cell r ' are sieved and then suspended in 5 ml of suspension medium and placed on filter paper through vacuum. The construct was coated into particles as is know in the art. The plates were then ~ ' ' ' The cells are then transferred to a N-6 medium and after 14 days II_~- F~ cells are selected by I mg/l bialaphos. The cells are then suspended in a medium containing .6% (w/v) (Sea-8 9 o Plaque; FMC) and held at 37C.
Two to five weeks later, growing calli are removed and transferred to the surface of fresh selection medium. Plants were ~ O ' in a MS based medium having 6% sucrose Ig/l myo-inositol, I mg/l NAA (34), and 0.3% (w/v) Gelrite (pH 6.0).
895 Next the embryo " cccurred in a MS media of 0.25 mgll NAA and 3%
(w/v) sucrose and light. Plants are grown and transferred to the O ' The expression levels in the plant can then be evaluated.
The plant is bred and developed to an inbred having the mutant and the down 900 reO~ulated pathway. Alternatively, the selected inbred can have the mutant crossed onto a transgenic after 1.. r ....- ~;.... to form the desired starch in the grain when the trdnsgenic plant is employed as the female.
Exampk 6 This example illustrates the i ' I . . of the starch of present invention compared to other starches. The O ' i . are listed in Table 3 below.

SUBSTITUTE SHEET (RULE 26) WO 95/35026 PCT/I~S9~/07828 2~ qQ~
910 Tsble3 h Samvles % Amylose~ T~ ture C
NatiYe common maize 2S 71 2 AMY V native 57 80 3 AMY Vll native 73 90 4 Present Invention 21 73 5 Native aewx 25 gO
6 Native wxwxwx 3 72 ~Values rounded to a whole number Sample I was a commercial product sold by American Maize-Products Company of Hammond, Indiana. The percent amylose and the ~ A~ for 915 Sample l above are mear~ values determined by random sampling of product. The 9g% confidence level for percent amylose and ~;, IA~ A~ I j r ' ~ are 25.9 to 29.3 and 68.7 to 72.9, ~
AMY V and AMY VII are commercial high amylose com starches sold by 920 American 1'~ -Pr. ' Company of Hammond, Indiana. The percent amylose and the t, l''; ~';'' , . in Table 3 above are mean values determined from a random sampling of product. The 99% confidence interval for the percent amylose in AMY V and AMY Vll was 53.4 to 62.5 and 65.5 to 73.8"~li~
The 99% confidence interval for the E,~ A~ -- i r ' for AMY V and 925 AMY vn was 72.8 to 84.4 and 83.1 to 90.8, .~li~ Both AMY V and AMY
Vll were grown in native maize.
Starch Sa~nple 4 w~ u~ to the Pr~sent Invention, while Sample 5 WII~IUIIJ~ to the average values from ~xample I of U. S. Patent No. 5,009,911.
930 Sample 6 cu.-. r to a commercial waxy starch sold by American Maize-Products Company.

SUBSTITUTE SHEET ~RULE 26) WO 95135026 2 1 9 3 7 6 1 r~l,o~)s. I~ ~
The method for ~ ~ botb the percent amylose and the g.
_ was:
935 The percent amylose ~vas determined using standard iodine procedures wherein the starch is first gelatinized with sodium hydroxide and then reacted with an iodine solution and the resulting sample measured using a ;"~11" ' in a I cm cell at 600 nm against a blank of 2% iodine solution.
940 The DSC ,, ' i , was measured using a scanning calorimeter ~ by Mettler Moddle No. 300 using 30% starch solids following the procedure outlined in the owner's manual for that model.
It is readily apparent from Table 3 above that the L ' : . _ of the 9 4 5 srarch of the present invention is comparable to common com starch.
Example 7 T.his example illustrates the gel strength of a sol made from 950 the com starch of the present invention compared to a sol made from aewx com starch, a sol made from common com starch, and a sol made from wxwxwx starch. The results of the test are reported in Table 4 below.
955 ~ Table 4 SamDle str~ne!h (grams) Present ~vention 159.5 Common Com Starch 225.0 aewx Com Starch 55.0 WaxyComStarch 16.0 SUBST~TU~E SHEET (RULE 2 2l 90761 In order to perform the gel strength test rcported in Table 4 above, sols were prepared by mixing water with starch and subjecting the slurry to a rapid heat mode in the Brabender Visco-Amylograph to heat the sample to 50 C. Once 50 C was 960 reached, the instrument was set at a controlled rate of heating, ~.5C1minute, until a of 95 C was reached. The sample was then held at 95 C for 30 minutes. Next, the sample was cooled at 1.5 C to a i . c of 50 C for 30 minutes. Portions of these sols were added separately to 4 ounce jars into which a plumger was placed. The sols were then allowed to stand at ambient conditions for 9 65 24 hours. Gcl strength was measured by ,:" ,, the force needed to remove the plunger from the sol.
This example illustrates that the gel strength of a sol made in accordance with the present invention is comparable to common com starch sols.

E:xample 8 This example illustrates the difference between aewx starch, a waxy starch wherein the plant had a triple dose of the 975 waxy gene, and the starch of the present invention. All starches were obtained from maize.
All starches were tested for their rheological prope~ties. Each starch was subjected to the same test procedure using the same method. The starch granules were pasting 9 8 0 using a Brabender Visco-Amylograph with the cooling probe down and with the 750 g cm cartridge. The starch slurry, 5.5% initial solids, was rapidly heated to 60 C in the Brabender cup, and then pasted while increasing the I . to 95 C at 1.5 C per minute. The starch paste was held at this t~ . for 20 minutes, and then ' 1~ loaded onto the measuring geometry of the rheometer which had been 9 8 5 preheated to 70 C. A four-patt rheological ~ -, was performed. The gel SUBSTITUTE SHEET ~RULE 26 woss~3so26 2 1 90 7 6 1 ~ C,07828 cure segment which monitored the formation of structure at .2 hertz and .2% strain (well within the linear viscoelastic region) was measured as the sample was cooled from 70 C to 25 C and held for 4 hours. Also messuled was the strain sweep, which measures the rheological response of the paste or gel to increasing levels of 9 9 0 strain, also at 02 hertz, at 25 C.
Using this technique, distinct differences between aewx starch and the starch ofthe present invention were found. Figure 5 shows the results of the gel cure analysis.
The starch of the present invention, while having an initial Modulus (G') lower than 9 9 5 aewx starch, more quickly formed structure or gelled as evidenced by the rapid rise in G'. Thus the starch of the Present Invention was distinct from aewx starch in the rate of gel formation despite similar apparent iodine binding contents. Figure 6 shows the results of the strain sweep analysis only for the starch of the present invention and the aew% starch. Here the starch of the present invention showed 10 0 0 dilatant behavior as evidenced by the increase in G' as the strain level was increased.
Under the applied levels of strain, the structure was not destroyed. In contrast, the structure of the ~ç~ starch was rapidly destroyed when the applied strain becamegreater than 4%. Thus, when compared to aewx starch, the starch of the present invention formed a gel which didn't break under the applied strain of this test. In 1005 contract, aewx starch showed much less time-dependent structure formation and did "break" or was destroyed by the strains applied in this technique.
Example g 1010 This example illustrates preparing a thickena ~ in accordance with the present invention.
The starch of the present invention is mixed with water in an amount to produce a slurry having 10% by weight starch. The sol has a short texture and a bland taste.
1015 The sol when cooked at about 90 C for lO minutes produces a thickener SUBSTlTUT~ SHEET (RULE 26 w095/3s026 21 q~7~ &
." which had better clarity than a similar thickener ~ ;.- . made from common com starch and a shorter texture.
Example 10 1020 ~
This example compares the mouth feel of a gel made from the starch of the present invention to a gel made with a common starch.
1025 The common starch and starch of the present invention were pasted using a Brabender visco-a~ . The starch was slurried at 5.5% solids, and then heated using the rapid heat mode to 50 C. Using controlled heat of 1.5 C per minute the slurries were heated to 95 C, and then held at this i , ~i for 30 minutes. The final solids was 5.9%. The sample starch pastes were then poured into small jelly 1030 jars, covered with cellophane, and allowed to age 24 hours before analysis. A taste panel was then asked to rank the samples for the following attributes.
First they ranked the two for relative firmness to the touch.
~Qmm~ Present ~vl~n~inn 5.1 5.6 Next, they ranked the relative break, firmness, and dearing of this sample whilebeing masticated.
Present Common ~j~
Degree of clean break 3.2 8.5 Firmness 4.4 4.5 Rate of clearing 2.5 4.6 SUBSTITUTE SHEET (RULE 26) wo gs/35026 2 1 9 0 7 6 1 PCTIUS95JO'1828 These results show that the firmness of these samples is similar. However the 10 4 0 starch of the present invention has a much cleaner break while being masticated.
r. I~ u~e~ it tends to clear from the mouth faster than a common based starch gel. The panel all agreed that the starch of the present invention produced a gel which had a "clean" mouth feel similar to that of a gelatin or a pectin.
10~5 Exsmple ll This example illustrates making a gum candy using the starch of the present invention.
10 5 0 The following ingredients and procedure are used:
Table 5 % by Weight In redients Present~v~.n~inn 44/62 Corn Syrup Unmixed 56.34 Sugar, fine granular 23.98 Water 7.73 P~sent Invention Starch l 1.80 Citric Acid Qo7 Sodium Citrate 0.08 100.00 Procedure 1055 All ingredients are mixed and then cooked to 340 F using ,UIl~,l..iUllal equipment such as a jet cooker. The cooked slurry is then poured into candy molds and allowed to solidify.

SUESTITUTE SHEET (RULE 26) woss/3so26 2.~ q~761 r~l,u .
Example 12 060 This example illustrates making a Bavarian cream pie using starch of the present invention.
The following ingredients and procedure are used:

Table 6 % by Weight In~redients Present ~vention Whole milk, fresh 3.5% 72.794 Sugar, fine grain ] 7.586 Salt, Flour QlOl Present Invention SAI0 Banana Flavoring 0.300 Egg Yolk, fresh 3.809 100.000 Procedure 1070 All of the pie filling ingredients except for egg yolks are combined and cooked at 195 F for 3 to 5 minutes. Then the ingredients are cooled to 120F with constant stirring. Next, egg yolks are added and the admixture well blended. This mixture is then added to a w~ iullal pie crust and allowed 1075 to cool to room i r ' before ser~ing.

SUBSTITUTE SHEET (RUL~ 2~) W09S/35026 2 1 9 0 7 6 1 PCTIUS9Sl07828 Example 13 This example illustrates preparing a lemon pie filling with the - 10 8 0 starch of the present invention.
The following ingredients and procedure are used:
Table 7 % By Weight In~redie~t~ p~cPnt InV~ntj~n Water 62.94 Sugar 19.30 ~'' ' 6.67 Present Invention 4.50 Corn Syrup Solids 2.50 Lemon Juice 2.50 Vegetable Shortening 1.03 Salt 0.23 Citric Acid 0.20 Emulsifier o.lo Lemon Oil (2x) 0.03 100.00 P~cedure Half of the water is combined with the sugar and brought to a boil. All of the remainmg ingredients are slurried together and then added to the boiling sugar and water. The 1090 ; , of this mixture is then adjusted to 200 F. and held there for two minutes. The mixture is then poured mto prepared pie crusts and allowed to cool and solidify.

SUBST~TUTE SHEET (RUL~

wo ss/3so26 r~m~
Example 14 10 9 5 This example illustrates making a chocolate mousse using the starch of the present invention. The formulation in Table g is employed to prepare a mousse mix.
Tllble X

%
Frodex 24-924 39.20 Sugar (Baker's) 30.75 Whiptreme 3554 (Kerry b~gredients) 12.88 Starch of Present Invention 9.80 Cocoa, Dutch Red (Gill .5c Duffus Products) 7.17 Leceitreme 40 (Kerry Ingredients) 0.20 Flavor as desired 100.00 Procedure Combine ingredients to form a uniform blend.
1105 Use Combine 200 gms. of mousse mix with l cup (250 grs.) milk. Using an electric mixer, combine on low speed for l minute. Scrape bowl. Mix on high speed for 3 minutes, until light and fluffy. Spoon into serving dishes and refrigerate for 1110 l hourbeforeserving.
To prepare the mousse mix, all the ingredients are mixed.
To prepare the mousse itself, 200 grams of mousse mix are SUBSTITUTE SHEET ~RULE 26) Wo 9513s026 1 ~ ~
~ 2 1 ~076 1 combined with 250 grams of milk and combined at a low 1115 speed. Then the mixture is sti~ed at a high speed to make it light and fluffy and the mixture is ,, ' ,, ' for one hour.
In this way, a light, fluffy mousse is prepared.
Accordingly, the present invention has been described with some degree of 112 0 I,~u Li~u L ;~y directed to the preferred ~ ' " of the p~sent invention. It should be Tr 1, though, that the present invention is defined by the following claimsconstrued in light of the prior art so that ,~ or changes may be made to the preferred e ' ' of the present invention without departing from the inventive concepts contained herein.

SU~STITUTE SHEET (RUL~ 2~)

Claims (35)

1. A plant comprising:
genomic material which includes genes which give incomplete reduction of the activity of at least two specific isoforms of the enzymes in the starch synthesis pathway of said plant.
2. A plant according to Claim 1 which forms starch which has different branchingstructure than the starch formed by a similar plant as described in Claim 1 which comprises genomic material which does not form isoforms of the enzymes in the starch synthesis pathway of the plant.
3. A plant according to Claim 2 in which said starch forms in grain.
4. Grain produced by a plant in which the genotype of the grain is mm*/n**, where m = first mutant n=second mutant and * equals wild type.
5. A starch producing plant comprising:
genomic material which includes genes which give incomplete reduction of the activity of at least two specific isoform enzymes in the starch synthesis pathway of said plant whereby said plant produces substantially more starch than said plant would produce if said genes gave complete reduction of the activity of thesame two specific isoform enzymes within the starch pathway.
6. Grain having an endosperm genotype selected from the group consisting of:
wxwxWx/AeAeae, aeaeAe/WxWxwx, wxwxWx/SuSusu, susuSu/wxWxwx, aeaeAe/SuSusu, susuSu/AeAeae, wxwxWx/DuDudu, aeaeAe/DuDudu, susuSu/DuDu du.
7. Grain having an endosperm genotype comprising two doses of a first mutant allele of a gene which affects starch structure and one dose of a second mutant allele of a second gene which affects starch structure, said genes can be selected from waxy, amylose extender, dull, horny, sugary, shrunken, brittle, floury, opaque.
8. The starch extracted from a grain according to claim 7 having a genotype of wxwxWx/AeAeae.
9. The starch extracted from a grain according to claim 7 having a genotype of Aeaeae/WxWxwx.
10. A plant having a diploid genotype of aa/BB and a triploid endosperm genotypeof aaA/BBb which has starch where a is a recessive mutant gene and A is the wild type gene, and b is a recessive mutant gene and B is the wild type gene andthe starch is altered from the normal starch, a and b can be selected from ae, du, wx, sh, bt, h, su, fl, op and B and A can be selected from Ae, Du, Wx, Sh, Bt, H, Su,F?,Op.
11. A pant having a diploid genotype of aA/Bb/Cc and a triploid endosperm genotype of aaA/BBb/CCc (and other combinations thereof) where a is a recessive mutant gene and A is the wild type gene, and b is a recessive mutant gene and B is the wild type gene, and c is a recessive mutant gene and C is the wild type gene and the starch is altered from the normal starch where a and b can be selected from ae, wx, sh, bt, h, su, fl, op, du and B and A can be selected from Ae, Wx, Sh, Bt, H, Su, Fl, Op, Du.
12. A method of producing grain with altered starch qualities which includes the steps of:

a) planting a parent which is capable of flowering and has complete reduction of at least one specific isoform enzyme in the starch synthesis pathway;
b) planting a second parent having a complete reduction of at least one other specific isoform enzyme in the starch synthesis pathway;
c) eliminating said first parent's capability to produce pollen;
d) pollinating said flowering first parent with said second parent's pollen;
and e) harvesting the grain produced by said first parent.
13. The method according to claim 12 including the step of extracting said altered starch from said grain.
14. The starch extracted from the grain according to claim 4 wherein a and b designate the same mutant and B and A designate the same wild type.
15. The starch extracted from grain having at least four doses of mutant and twodoses of wild type, such that the genotype has wild type on each side.
16. The starch extracted from grain having at least three doses of mutant and three doses of wild type, such that the genotype has mutation on each side.
17. A single mutant male sterile plant where the mutant is selected from ae, wx, sh, bt, h, su, fl, op, du.
18. A sol comprising water and an effective amount of a starch extracted from a starch bearing plant having a waxy, waxy, amylose extender genotype.
19. The sol of claim 18 wherein the starch is present in an amount of about 1% to about 20% by weight.
20. The sol of claim 18 wherein the plant is maize.
21. The sol of claim 18 wherein the starch is cold water soluble.
22. The sol of claim 18 wherein said starch is in granular form.
23. The sol of claim 21 wherein said starch is extracted from maize.
24. A foodstuff comprising a foodstubb and having as an essential ingredient therein an effective amount of a starch extracted from a starch bearing plant having a waxy, waxy, amylose extender genotype.
25. The foodstuff of claim 24 wherein said starch is present in an amount of about 0.1% to about 10% by weight foodstuff.
26. The foodstuff of claim 24 wherein said starch is extracted from maize.
27. A method for making a sol containing starch comprising the steps of:
forming a slurry comprising water and an effective amount of a starch extracted from a starch bearing plant having waxy, waxy, amylose genotype; and cooking said starch to gelatinize said starch.
28. The method of claim 27 wherein said effective amount is about 1% to about 20% by weight slurry.
29. The method of claim 27 wherein said starch is extracted from maize.
30. A method for thickening a foodstuff comprising the steps of:
combining with a foodstuff an effective amount of a starch extracted from a starch bearing plant having a waxy, waxy, amylose extender genotype; and cooking said foodstuff to thicken said foodstuff.
31. The method of claim 30 wherein said starch is extracted from maize.
32. The method of claim 30 wherein said starch is present in an amount of from about 0.1% to about 10% by weight of said foodstuff.
33. An improved method for making a foodstuff which contains a gelatin said improvement comprising replacing at least a portion of the gelatin in said foodstuff with the sol of claim 18.
34. An improved method for making a foodstuff which contains a natural gum, saidimprovement comprising replacing at least a portion of the natural gum with the sol of claim 18.
35. The method of claim 34 wherein said foodstuff is a gum candy, a gelled dessert, a glaze, or a spread.
CA002190761A 1994-06-21 1995-06-20 Novel plants and processes for obtaining them Abandoned CA2190761A1 (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
US26392194A true 1994-06-21 1994-06-21
US08/263,921 1994-06-21
US34660294A true 1994-11-29 1994-11-29
US08/346,602 1994-11-29
US47406395A true 1995-06-07 1995-06-07
US08/487,466 1995-06-07
US08/487,466 US5576048A (en) 1995-06-07 1995-06-07 Foodstuffs containing a waxy waxy amylose extender starch
US08/474,063 1995-06-07

Publications (1)

Publication Number Publication Date
CA2190761A1 true CA2190761A1 (en) 1995-12-28

Family

ID=27500804

Family Applications (1)

Application Number Title Priority Date Filing Date
CA002190761A Abandoned CA2190761A1 (en) 1994-06-21 1995-06-20 Novel plants and processes for obtaining them

Country Status (6)

Country Link
EP (1) EP0802720A4 (en)
JP (2) JPH10507622A (en)
CN (1) CN1156951A (en)
AU (1) AU706849B2 (en)
CA (1) CA2190761A1 (en)
WO (1) WO1995035026A1 (en)

Families Citing this family (174)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9705746D0 (en) * 1997-03-20 1997-05-07 Spp Food rheology improvements
CA2295931C (en) 1997-06-26 2010-05-18 Sapporo Breweries Ltd. A method for identifying a barley variety and a barley having a brewing property
AU758890B2 (en) * 1998-06-15 2003-04-03 National Starch And Chemical Investment Holding Corporation Improvements in or relating to plants and plant products
CA2416632C (en) * 2000-08-28 2012-01-24 E.I. Du Pont De Nemours And Company Novel starches produced by the expression of heterologous granule bound starch synthase genes
US7169982B2 (en) * 2001-10-17 2007-01-30 Basf Plant Science Gmbh Starch
US6960703B2 (en) 2002-03-06 2005-11-01 National Starch And Chemical Investment Holdings Corporation Grain production method for maize starch with novel functionality
US6828474B2 (en) 2002-03-06 2004-12-07 National Starch & Chemical Investment Holding Corporation Method of grain production for heterozygous waxy sugary-2 maize
JP4695471B2 (en) * 2005-09-16 2011-06-08 松谷化学工業株式会社 Soft candy and a method of manufacturing the same
CL2007003744A1 (en) 2006-12-22 2008-07-11 Bayer Cropscience Ag Composition comprising a derivative 2-pyridylmethylbenzamide and an insecticide compound; and method for curatively controlling phytopathogenic fungi or preventive crop and insects.
EP1969934A1 (en) 2007-03-12 2008-09-17 Bayer CropScience AG 4-cycloalkyl or 4-aryl substituted phenoxy phenylamidines and their use as fungicides
EP1969930A1 (en) 2007-03-12 2008-09-17 Bayer CropScience AG Phenoxy phenylamidines and their use as fungicides
EP1969929A1 (en) 2007-03-12 2008-09-17 Bayer CropScience AG Substituted phenylamidines and their use as fungicides
EP2136627B1 (en) 2007-03-12 2015-05-13 Bayer Intellectual Property GmbH Dihalophenoxyphenylamidines and use thereof as fungicides
WO2008110280A2 (en) 2007-03-12 2008-09-18 Bayer Cropscience Ag Phenoxy substituted phenylamidine derivatives and their use as fungicides
DE102007045953B4 (en) 2007-09-26 2018-07-05 Bayer Intellectual Property Gmbh Drug combinations with insecticidal and acaricidal properties
DE102007045920B4 (en) 2007-09-26 2018-07-05 Bayer Intellectual Property Gmbh Synergistic combinations of active substances
DE102007045919B4 (en) 2007-09-26 2018-07-05 Bayer Intellectual Property Gmbh Drug combinations with insecticidal and acaricidal properties
DE102007045922A1 (en) 2007-09-26 2009-04-02 Bayer Cropscience Ag Drug combinations with insecticidal and acaricidal properties
EP2072506A1 (en) 2007-12-21 2009-06-24 Bayer CropScience AG Thiazolyloxyphenylamidine or thiadiazolyloxyphenylamidine und its use as fungicide
EP2090168A1 (en) 2008-02-12 2009-08-19 Bayer CropScience AG Method for improving plant growth
EP2168434A1 (en) 2008-08-02 2010-03-31 Bayer CropScience AG Use of azols to increase resistance of plants of parts of plants to abiotic stress
PE06722011A1 (en) 2008-08-14 2011-09-25 Bayer Cropscience Ag 4-phenyl-1-H-pyrazoles insecticides
DE102008041695A1 (en) 2008-08-29 2010-03-04 Bayer Cropscience Ag Methods to improve plant growth
EP2201838A1 (en) 2008-12-05 2010-06-30 Bayer CropScience AG Active ingredient-beneficial organism combinations with insecticide and acaricide properties
EP2198709A1 (en) 2008-12-19 2010-06-23 Bayer CropScience AG Method for treating resistant animal pests
EP2204094A1 (en) 2008-12-29 2010-07-07 Bayer CropScience AG Method for improved utilization of the production potential of transgenic plants Introduction
US9763451B2 (en) 2008-12-29 2017-09-19 Bayer Intellectual Property Gmbh Method for improved use of the production potential of genetically modified plants
EP2039772A2 (en) 2009-01-06 2009-03-25 Bayer CropScience AG Method for improved utilization of the production potential of transgenic plants introduction
EP2039770A2 (en) 2009-01-06 2009-03-25 Bayer CropScience AG Method for improved utilization of the production potential of transgenic plants
EP2039771A2 (en) 2009-01-06 2009-03-25 Bayer CropScience AG Method for improved utilization of the production potential of transgenic plants
JP5558490B2 (en) 2009-01-19 2014-07-23 バイエル・クロップサイエンス・アーゲーBayer Cropscience Ag Cyclic dione and pesticides, their use as acaricides and / or fungicides
EP2227951A1 (en) 2009-01-23 2010-09-15 Bayer CropScience AG Application of enaminocarbonyl compounds for combating viruses transmitted by insects
CN102300852B (en) 2009-01-28 2015-04-22 拜尔农科股份公司 Fungicide N-cycloalkyl-N-bicyclicmethylene-carboxamide derivatives
AR075126A1 (en) 2009-01-29 2011-03-09 Bayer Cropscience Ag Method for the best use of the potential production of transgenic plants
CN102317259B (en) 2009-02-17 2015-12-02 拜尔农科股份公司 Fungicidal n- (phenyl cycloalkyl) carboxamide, n- (benzyl cycloalkyl) carboxamide and thiocarboxamide derivative
EP2218717A1 (en) 2009-02-17 2010-08-18 Bayer CropScience AG Fungicidal N-((HET)Arylethyl)thiocarboxamide derivatives
TW201031331A (en) 2009-02-19 2010-09-01 Bayer Cropscience Ag Pesticide composition comprising a tetrazolyloxime derivative and a fungicide or an insecticide active substance
EP2223602A1 (en) 2009-02-23 2010-09-01 Bayer CropScience AG Method for improved utilisation of the production potential of genetically modified plants
EP2232995A1 (en) 2009-03-25 2010-09-29 Bayer CropScience AG Method for improved utilisation of the production potential of transgenic plants
WO2010108508A2 (en) 2009-03-25 2010-09-30 Bayer Cropscience Ag Active ingredient combinations with insecticidal and acaricidal properties
US8828906B2 (en) 2009-03-25 2014-09-09 Bayer Cropscience Ag Active compound combinations having insecticidal and acaricidal properties
EP2410847A1 (en) 2009-03-25 2012-02-01 Bayer CropScience AG Active ingredient combinations having insecticidal and acaricidal properties
WO2010108507A2 (en) 2009-03-25 2010-09-30 Bayer Cropscience Ag Synergistic combinations of active ingredients
EP2239331A1 (en) 2009-04-07 2010-10-13 Bayer CropScience AG Method for improved utilization of the production potential of transgenic plants
BRPI1015543A2 (en) 2009-05-06 2015-09-01 Bayer Cropscience Ag cyclopentanedione compounds and their use as insecticides, acaricides and / or fungicides.
EP2251331A1 (en) 2009-05-15 2010-11-17 Bayer CropScience AG Fungicide pyrazole carboxamides derivatives
AR076839A1 (en) 2009-05-15 2011-07-13 Bayer Cropscience Ag Pyrazole carboxamides derivatives fungicides
EP2255626A1 (en) 2009-05-27 2010-12-01 Bayer CropScience AG Use of succinate dehydrogenase inhibitors to increase resistance of plants or parts of plants to abiotic stress
CN102595889A (en) 2009-06-02 2012-07-18 拜耳作物科学公司 Use of succinate dehydrogenase inhibitors for controlling sclerotinia ssp.
AU2010272872B2 (en) 2009-07-16 2014-08-28 Bayer Intellectual Property Gmbh Synergistic active substance combinations containing phenyl triazoles
JP5641179B2 (en) * 2009-07-22 2014-12-17 公立大学法人大阪府立大学 Penetration substance holding cereals
WO2011015524A2 (en) 2009-08-03 2011-02-10 Bayer Cropscience Ag Fungicide heterocycles derivatives
EP2292094A1 (en) 2009-09-02 2011-03-09 Bayer CropScience AG Active compound combinations
EP2343280A1 (en) 2009-12-10 2011-07-13 Bayer CropScience AG Fungicide quinoline derivatives
TWI528898B (en) 2009-12-28 2016-04-11 Bayer Ip Gmbh Fungicide hydroximoyl-heterocycles derivatives
WO2011080256A1 (en) 2009-12-28 2011-07-07 Bayer Cropscience Ag Fungicide hydroximoyl-tetrazole derivatives
JP5782657B2 (en) 2009-12-28 2015-09-24 バイエル・クロップサイエンス・アクチェンゲゼルシャフト Fungicides hydroximoyl - tetrazole derivatives
NZ601341A (en) 2010-01-22 2014-02-28 Bayer Ip Gmbh Acaricide and/or insecticide active substance combinations
US20110218103A1 (en) 2010-03-04 2011-09-08 Bayer Cropscience Ag Fluoroalkyl-substituted 2-amidobenzimidazoles
JP2013523795A (en) 2010-04-06 2013-06-17 バイエル・インテレクチユアル・プロパテイー・ゲー・エム・ベー・ハー The use of 4-phenylbutyric acid and / or salts thereof for enhancing the stress tolerance of a plant
CN102933083B (en) 2010-04-09 2015-08-12 拜耳知识产权有限责任公司 (1-cyano-cyclopropyl) phenyl phosphinic acid derivative or an ester thereof and / or a salt thereof to improve the use of plants to abiotic stress tolerance
EP2563772A1 (en) 2010-04-28 2013-03-06 Bayer Cropscience AG Fungicide hydroximoyl-heterocycles derivatives
US20130116287A1 (en) 2010-04-28 2013-05-09 Christian Beier Fungicide hydroximoyl-heterocycles derivatives
WO2011134911A2 (en) 2010-04-28 2011-11-03 Bayer Cropscience Ag Fungicide hydroximoyl-tetrazole derivatives
KR20130109940A (en) 2010-06-03 2013-10-08 바이엘 크롭사이언스 아게 N-[(het)arylethyl] pyrazole(thio)carboxamides and their heterosubstituted analogues
KR20130088022A (en) 2010-06-03 2013-08-07 바이엘 크롭사이언스 아게 N-[(het)arylalkyl] pyrazole(thio)carboxamides and their heterosubstituted analogues
AU2011260331B2 (en) 2010-06-03 2014-04-17 Bayer Cropscience Ag Fungicide N-[(trisubstitutedsilyl)methyl]-carboxamide derivatives
CN103119169B (en) 2010-06-09 2018-11-20 拜尔作物科学公司 Transformation of the plant genome and commonly used method of modifying the plant genome of tools in nucleotide sequence
AU2011264074B2 (en) 2010-06-09 2015-01-22 Bayer Cropscience Nv Methods and means to modify a plant genome at a nucleotide sequence commonly used in plant genome engineering
EP2595961B1 (en) 2010-07-20 2017-07-19 Bayer Intellectual Property GmbH Benzocycloalkenes as antifungal agents
CN103228141B (en) 2010-09-03 2016-04-20 拜耳知识产权有限责任公司 Substituted fused pyrimidinone and dihydropyrimidinone
CN103298341B (en) 2010-09-22 2016-06-08 拜耳知识产权有限责任公司 Active ingredient for use in controlling nematodes nematode resistant crops
RS58401B1 (en) 2010-10-07 2019-04-30 Bayer Cropscience Ag Fungicide composition comprising a tetrazolyloxime derivative and a thiazolylpiperidine derivative
EP2630135A1 (en) 2010-10-21 2013-08-28 Bayer Intellectual Property GmbH 1-(heterocyclic carbonyl) piperidines
UA107865C2 (en) 2010-10-21 2015-02-25 Байєр Інтелекчуал Проперті Гмбх Heterocyclic carboxamides
MX2013004878A (en) 2010-11-02 2013-07-02 Bayer Ip Gmbh N-hetarylmethyl pyrazolylcarboxamides.
AR083874A1 (en) 2010-11-15 2013-03-27 Bayer Cropscience Ag 5-halogenopirazol (thio) carboxamides
WO2012065944A1 (en) 2010-11-15 2012-05-24 Bayer Cropscience Ag N-aryl pyrazole(thio)carboxamides
JP5833663B2 (en) 2010-11-15 2015-12-16 バイエル・インテレクチュアル・プロパティ・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツングBayer Intellectual Property GmbH 5-halogeno-pyrazole carboxamides
EP3372081A3 (en) 2010-12-01 2018-10-24 Bayer CropScience Aktiengesellschaft Use of fluopyram for controlling nematodes in crops
EP2460406A1 (en) 2010-12-01 2012-06-06 Bayer CropScience AG Use of fluopyram for controlling nematodes in nematode resistant crops
EP2460407A1 (en) 2010-12-01 2012-06-06 Bayer CropScience AG Agent combinations comprising pyridylethyl benzamides and other agents
BR112013016755A2 (en) 2010-12-29 2016-07-12 Bayer Intelectual Property Gmbh tetrazoiloxima derivative of formula (I) compound and method for controlling phytopathogenic fungi of crops
EP2474542A1 (en) 2010-12-29 2012-07-11 Bayer CropScience AG Fungicide hydroximoyl-tetrazole derivatives
EP2471363A1 (en) 2010-12-30 2012-07-04 Bayer CropScience AG Use of aryl-, heteroaryl- and benzylsulfonamide carboxylic acids, -carboxylic acid esters, -carboxylic acid amides and -carbonitriles and/or its salts for increasing stress tolerance in plants
EP2494867A1 (en) 2011-03-01 2012-09-05 Bayer CropScience AG Halogen-substituted compounds in combination with fungicides
EP2683239A1 (en) 2011-03-10 2014-01-15 Bayer Intellectual Property GmbH Use of lipochito-oligosaccharide compounds for safeguarding seed safety of treated seeds
JP2014509599A (en) 2011-03-14 2014-04-21 バイエル・インテレクチユアル・プロパテイー・ゲー・エム・ベー・ハー Fungicides hydroximoyl - tetrazole derivatives
BR112013025871A2 (en) 2011-04-08 2016-07-26 Bayer Ip Gmbh compound of formula (I) and their use composition for the control of phytopathogenic harmful fungi, a method for controlling phytopathogenic fungi of crops and the process for making the compositions
AR090010A1 (en) 2011-04-15 2014-10-15 Bayer Cropscience Ag 5- (cyclohex-2-en-1-yl) penta-2,4-dienes and 5- (cyclohex-2-en-1-yl) pent-2-en-4-ynes substituted as active against abiotic stress in plants, uses and methods of treatment
EP2511255A1 (en) 2011-04-15 2012-10-17 Bayer CropScience AG Substituted prop-2-in-1-ol and prop-2-en-1-ol derivatives
AR085568A1 (en) 2011-04-15 2013-10-09 Bayer Cropscience Ag 5- (bicyclo [4.1.0] hept-3-en-2-yl) penta-2,4-dienes and 5- (bicyclo [4.1.0] hept-3-en-2-yl) -pent- 2-en-4-ynes substituted as active against plant abiotic stress
AR085585A1 (en) 2011-04-15 2013-10-09 Bayer Cropscience Ag Vinyl- and alquinilciclohexanoles substituted as active against plant abiotic stress
BR112013027091A8 (en) 2011-04-22 2018-01-16 Bayer Ip Gmbh "Combination of the active compound composition for controlling phytopathogenic harmful fungi, the method for controlling phytopathogenic harmful fungi, a process for producing compositions for controlling phytopathogenic harmful fungi and uses a combination of the active compound"
US20140173770A1 (en) 2011-06-06 2014-06-19 Bayer Cropscience Nv Methods and means to modify a plant genome at a preselected site
WO2013004652A1 (en) 2011-07-04 2013-01-10 Bayer Intellectual Property Gmbh Use of substituted isoquinolinones, isoquinolindiones, isoquinolintriones and dihydroisoquinolinones or in each case salts thereof as active agents against abiotic stress in plants
CN103717076B (en) 2011-08-10 2016-04-13 拜耳知识产权股份有限公司 Active compound composition containing the specific tetramic acid derivatives
JP2014524455A (en) 2011-08-22 2014-09-22 バイエル・インテレクチユアル・プロパテイー・ゲー・エム・ベー・ハー Fungicidal hydroximoyl - tetrazole derivatives
CN103981149A (en) 2011-08-22 2014-08-13 拜尔作物科学公司 Methods and means to modify a plant genome
EP2561759A1 (en) 2011-08-26 2013-02-27 Bayer Cropscience AG Fluoroalkyl-substituted 2-amidobenzimidazoles and their effect on plant growth
EP2753177A1 (en) 2011-09-09 2014-07-16 Bayer Intellectual Property GmbH Acyl-homoserine lactone derivatives for improving plant yield
US9090600B2 (en) 2011-09-12 2015-07-28 Bayer Intellectual Property Gmbh Fungicidal 4-substituted-3-{phenyl[(heterocyclylmethoxy)imino]methyl}-1,2,4-oxadizol-5(4H)-one derivatives
CN108552186A (en) 2011-09-16 2018-09-21 拜耳知识产权有限责任公司 Use of phenylpyrazolin-3-carboxylates for improving plant yield
WO2013037955A1 (en) 2011-09-16 2013-03-21 Bayer Intellectual Property Gmbh Use of acylsulfonamides for improving plant yield
MX357718B (en) 2011-09-16 2018-07-20 Bayer Ip Gmbh Use of 5-phenyl- or 5-benzyl-2 isoxazoline-3 carboxylates for improving plant yield.
JP2014527973A (en) 2011-09-23 2014-10-23 バイエル・インテレクチユアル・プロパテイー・ゲー・エム・ベー・ハー The use of 4-substituted 1-phenyl-pyrazole-3-carboxylic acid derivatives as agents to abiotic plant stresses
CA2844868A1 (en) 2011-10-04 2013-04-11 Bayer Intellectual Property Gmbh Rnai for the control of fungi and oomycetes by inhibiting saccharopine dehydrogenase gene
WO2013050324A1 (en) 2011-10-06 2013-04-11 Bayer Intellectual Property Gmbh Combination, containing 4-phenylbutyric acid (4-pba) or a salt thereof (component (a)) and one or more selected additional agronomically active compounds (component(s) (b)), that reduces abiotic plant stress
US9617286B2 (en) 2011-11-21 2017-04-11 Bayer Intellectual Property Gmbh Fungicide N-[(trisubstitutedsilyl)methyl]-carboxamide derivatives
US9725414B2 (en) 2011-11-30 2017-08-08 Bayer Intellectual Property Gmbh Fungicidal N-bicycloalkyl and N-tricycloalkyl pyrazole-4-(thio)carboxamide derivatives
WO2013092519A1 (en) 2011-12-19 2013-06-27 Bayer Cropscience Ag Use of anthranilic acid diamide derivatives for pest control in transgenic crops
EP2797891B1 (en) 2011-12-29 2015-09-30 Bayer Intellectual Property GmbH Fungicidal 3-[(pyridin-2-ylmethoxyimino)(phenyl)methyl]-2-substituted-1,2,4-oxadiazol-5(2h)-one derivatives
EP2797895B1 (en) 2011-12-29 2015-08-05 Bayer Intellectual Property GmbH Fungicidal 3-[(1,3-thiazol-4-ylmethoxyimino)(phenyl)methyl]-2-substituted-1,2,4-oxadiazol-5(2h)-one derivatives
EP2816897B1 (en) 2012-02-22 2018-02-07 Bayer CropScience AG Use of fluopyram for controlling wood diseases in grape
UA113198C2 (en) 2012-02-27 2016-12-26 Combinations of active compounds
WO2013139949A1 (en) 2012-03-23 2013-09-26 Bayer Intellectual Property Gmbh Compositions comprising a strigolactame compound for enhanced plant growth and yield
EP2836489B1 (en) 2012-04-12 2016-06-29 Bayer Cropscience AG N-acyl-2-(cyclo) alkylpyrrolidines and piperidines useful as fungicides
EP2838363A1 (en) 2012-04-20 2015-02-25 Bayer Cropscience AG N-cycloalkyl-n-[(trisubstitutedsilylphenyl)methylene]-(thio)carboxamide derivatives
WO2013156559A1 (en) 2012-04-20 2013-10-24 Bayer Cropscience Ag N-cycloalkyl-n-[(heterocyclylphenyl)methylene]-(thio)carboxamide derivatives
CN104245940A (en) 2012-04-23 2014-12-24 拜尔作物科学公司 Targeted genome engineering in plants
EP2662364A1 (en) 2012-05-09 2013-11-13 Bayer CropScience AG Pyrazole tetrahydronaphthyl carboxamides
EP2662361A1 (en) 2012-05-09 2013-11-13 Bayer CropScience AG Pyrazol indanyl carboxamides
EP2662362A1 (en) 2012-05-09 2013-11-13 Bayer CropScience AG Pyrazole indanyl carboxamides
EP2662360A1 (en) 2012-05-09 2013-11-13 Bayer CropScience AG 5-Halogenopyrazole indanyl carboxamides
EP2662370A1 (en) 2012-05-09 2013-11-13 Bayer CropScience AG 5-Halogenopyrazole benzofuranyl carboxamides
CN104768934B (en) 2012-05-09 2017-11-28 拜耳农作物科学股份公司 Pyrazol-indanyl carboxamide
US9375005B2 (en) 2012-05-09 2016-06-28 Bayer Cropscience Ag 5-halogenopyrazole indanyl carboxamides
EP2662363A1 (en) 2012-05-09 2013-11-13 Bayer CropScience AG 5-Halogenopyrazole biphenylcarboxamides
AR091104A1 (en) 2012-05-22 2015-01-14 Bayer Cropscience Ag Combinations of active compounds comprising a lipo-chito-oligosaccharide derivative and a compound nematicide, insecticide or fungicide
AU2013289301A1 (en) 2012-07-11 2015-01-22 Bayer Cropscience Ag Use of fungicidal combinations for increasing the tolerance of a plant towards abiotic stress
AU2013311826A1 (en) 2012-09-05 2015-03-26 Bayer Cropscience Ag Use of substituted 2-amidobenzimidazoles, 2-amidobenzoxazoles and 2-amidobenzothiazoles or salts thereof as active substances against abiotic plant stress
EA026839B1 (en) 2012-10-19 2017-05-31 Байер Кропсайенс Аг Active compound combinations comprising carboxamide compounds
EA025669B1 (en) 2012-10-19 2017-01-30 Байер Кропсайенс Аг Method of plant growth promotion using carboxamide derivatives
US9668480B2 (en) 2012-10-19 2017-06-06 Bayer Cropscience Ag Method for treating plants against fungi resistant to fungicides using carboxamide or thiocarboxamide derivatives
JP6184502B2 (en) 2012-10-19 2017-08-23 バイエル・クロップサイエンス・アクチェンゲゼルシャフト Method of enhancing tolerance to abiotic stress in plants using carboxamide or thiocarboxamide derivatives
WO2014063442A1 (en) * 2012-10-23 2014-05-01 中国农业大学 Method for propagating sterile male plant line
CN102965391B (en) * 2012-10-23 2014-09-24 中国农业大学 High-efficiency seed labeling method for propagation of plant male sterile line
EP2735231A1 (en) 2012-11-23 2014-05-28 Bayer CropScience AG Active compound combinations
WO2014079957A1 (en) 2012-11-23 2014-05-30 Bayer Cropscience Ag Selective inhibition of ethylene signal transduction
UA117819C2 (en) 2012-11-30 2018-10-10 Байєр Кропсайєнс Акцієнгезелльшафт Binary pesticidal and fungicidal mixtures
MX2015006327A (en) 2012-11-30 2015-10-05 Bayer Cropscience Ag Ternary fungicidal mixtures.
UA117820C2 (en) 2012-11-30 2018-10-10 Байєр Кропсайєнс Акцієнгезелльшафт Binary fungicidal or pesticidal mixture
CN104812247A (en) 2012-11-30 2015-07-29 拜耳作物科学股份公司 Binary fungicidal mixtures
CN104918493B (en) 2012-11-30 2018-02-06 拜尔农作物科学股份公司 Three yuan fungicidal and insecticidal mixture
EP2740720A1 (en) 2012-12-05 2014-06-11 Bayer CropScience AG Substituted bicyclic and tricyclic pent-2-en-4-inic acid derivatives and their use for enhancing the stress tolerance in plants
BR112015012926A2 (en) 2012-12-05 2017-07-11 Bayer Cropscience Ag Use of 1- (aryl ethynyl) - 1- (ethynyl heteroaryl) -, 1- (ethynyl heterocyclyl) - substituted and 1- (ethynyl cycloalkenyl) -ciclohexanóis as active agents against plant abiotic stress
EP2740356A1 (en) 2012-12-05 2014-06-11 Bayer CropScience AG Substituted (2Z)-5(1-Hydroxycyclohexyl)pent-2-en-4-inic acid derivatives
WO2014090765A1 (en) 2012-12-12 2014-06-19 Bayer Cropscience Ag Use of 1-[2-fluoro-4-methyl-5-(2,2,2-trifluoroethylsulfinyl)phenyl]-5-amino-3-trifluoromethyl)-1 h-1,2,4 tfia zole for controlling nematodes in nematode-resistant crops
AR093996A1 (en) 2012-12-18 2015-07-01 Bayer Cropscience Ag bactericides fungicides and binary logic
CN104995174A (en) 2012-12-19 2015-10-21 拜耳作物科学股份公司 Difluoromethyl-nicotinic-tetrahydronaphtyl carboxamides
TW201446759A (en) 2013-03-07 2014-12-16 Bayer Cropscience Ag Fungicidal 3-{phenyl[(heterocyclylmethoxy)imino]methyl}-heterocycle derivatives
BR112015025006A2 (en) 2013-04-02 2017-10-10 Bayer Cropscience Nv genomic engineering targeted in eukaryotes
WO2014167008A1 (en) 2013-04-12 2014-10-16 Bayer Cropscience Ag Novel triazolinthione derivatives
EP2984081B1 (en) 2013-04-12 2017-08-09 Bayer CropScience Aktiengesellschaft Novel triazole derivatives
EP2986117A1 (en) 2013-04-19 2016-02-24 Bayer CropScience Aktiengesellschaft Binary insecticidal or pesticidal mixture
US20160058001A1 (en) 2013-04-19 2016-03-03 Bayer Cropscience Aktiengesellschaft Method for improved utilization of the production potential of transgenic plants
TW201507722A (en) 2013-04-30 2015-03-01 Bayer Cropscience Ag N-(2-halogen-2-phenethyl)carboxamides as nematicides and endoparasiticides
WO2014177514A1 (en) 2013-04-30 2014-11-06 Bayer Cropscience Ag Nematicidal n-substituted phenethylcarboxamides
BR112015031235A2 (en) 2013-06-26 2017-07-25 Bayer Cropscience Ag derivatives of N-cycloalkyl-N - [(biciclil-phenyl) methylene] - (thio) carboxamide
EA201600097A1 (en) 2013-07-09 2016-06-30 Байер Кропсайенс Акциенгезельшафт Application of selected pyridon carboxamides or their salts as active substances against the abiotic stress of plants
ES2705577T3 (en) 2013-12-05 2019-03-26 Bayer Cropscience Ag Derivatives of N-cyclopropyl-N - {[2- (1-cyclopropyl substituted) phenyl] methylene} - (thio) carboxamide
CN105873907B (en) 2013-12-05 2019-03-12 拜耳作物科学股份公司 N- naphthenic base-N- { [2- (naphthenic base that 1- replaces) phenyl] methylene }-(thio) carboxamides derivatives
AR101214A1 (en) 2014-07-22 2016-11-30 Bayer Cropscience Ag Cyano-2,4-dienes cicloalquilpenta-, cyano-cicloalquilpent-2-en-4-inas, cyano-2,4-dienes heterociclilpenta-cyano-heterociclilpent and 2-en-4-inas substituted as active anti-stress abiotic plant
AR103024A1 (en) 2014-12-18 2017-04-12 Bayer Cropscience Ag selected Piridoncarboxamidas or their salts as active substances against abiotic stress of plants
EP3283476B1 (en) 2015-04-13 2019-08-14 Bayer Cropscience AG N-cycloalkyl-n-(biheterocyclyethylene)-(thio)carboxamide derivatives
BR112019001764A2 (en) 2016-07-29 2019-05-07 Bayer Cropscience Ag combinations of active compounds and methods for plant propagation material protecting
EP3515907A1 (en) 2016-09-22 2019-07-31 Bayer CropScience Aktiengesellschaft Novel triazole derivatives
BR112019005660A2 (en) 2016-09-22 2019-06-04 Bayer Cropscience Ag new triazole derivatives and their use as fungicides
US20190225974A1 (en) 2016-09-23 2019-07-25 BASF Agricultural Solutions Seed US LLC Targeted genome optimization in plants
CN109890204A (en) 2016-10-26 2019-06-14 拜耳作物科学股份公司 Pyraziflumid is used to control the purposes of Sclerotinia kind in seed treatment application
CA3046145A1 (en) 2016-12-08 2018-06-14 Bayer Cropscience Aktiengesellschaft Use of insecticides for controlling wireworms
EP3332645A1 (en) 2016-12-12 2018-06-13 Bayer Cropscience AG Use of substituted pyrimidine diones or their salts as agents to combat abiotic plant stress
WO2018108627A1 (en) 2016-12-12 2018-06-21 Bayer Cropscience Aktiengesellschaft Use of substituted indolinylmethyl sulfonamides, or the salts thereof for increasing the stress tolerance of plants
WO2019025153A1 (en) 2017-07-31 2019-02-07 Bayer Cropscience Aktiengesellschaft Use of substituted n-sulfonyl-n'-aryl diaminoalkanes and n-sulfonyl-n'-heteroaryl diaminoalkanes or salts thereof for increasing the stress tolerance in plants

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4770710A (en) * 1987-07-02 1988-09-13 American Maize-Products Company Novel starch and products produced therefrom
US4792458A (en) * 1987-07-02 1988-12-20 American Maize-Products Company Food stuffs containing starch of a dull sugary-2 genotype
US4798735A (en) * 1987-07-02 1989-01-17 American Maize-Products Company Foodstuffs containing starch of an amylose extender sugary-2 genotype
US4790997A (en) * 1987-07-02 1988-12-13 American Maize-Products Company Food stuffs containing starch of an amylose extender dull genotype
US4767849A (en) * 1987-07-02 1988-08-30 American Maize-Products Company Starch of the wxsh1 genotype and products produced therefrom
US5004864A (en) * 1988-11-28 1991-04-02 Iowa State University Research Foundation, Inc. Dominant amylose-extender mutant of maize
AU631403B2 (en) * 1989-11-20 1992-11-26 National Starch Llc Novel maize products
US5502270A (en) * 1993-03-30 1996-03-26 E. I. Du Pont De Nemours And Company Starch and grain with a novel genotype
JP5241158B2 (en) * 2007-07-10 2013-07-17 キヤノン株式会社 Image forming apparatus

Also Published As

Publication number Publication date
EP0802720A1 (en) 1997-10-29
EP0802720A4 (en) 1999-01-13
JPH10507622A (en) 1998-07-28
AU2905395A (en) 1996-01-15
JP2007267740A (en) 2007-10-18
CN1156951A (en) 1997-08-13
AU706849B2 (en) 1999-06-24
WO1995035026A1 (en) 1995-12-28

Similar Documents

Publication Publication Date Title
Albrecht et al. Cell-Wall Composition and Digestibility of Alfalfa Stems and Leaves 1
CA2421679C (en) Monocotyledon plant cells and plants which synthesise modified starch
US6255561B1 (en) Nucleic acid molecules coding for debranching enzymes from maize
CN100509853C (en) Starch
US6897358B2 (en) Nucleic acid molecules encoding wheat enzymes involved in starch synthesis
DE69637153T2 (en) Improvements in or relating to pfanzenstärkeverbindungen
AU730569B2 (en) Nucleic acid molecules encoding starch phosphorylase from maize
AU715944B2 (en) Plants which synthesize a modified starch, process for the production thereof and modified starch
Peña Wheat for bread and other foods
EP0791066B1 (en) Dna molecules that code for enzymes involved in starch synthesis, vectors, bacteria, transgenic plant cells and plants containing said molecules
Belderok et al. Bread-making quality of wheat: a century of breeding in Europe
US20030074685A1 (en) Soybean plant producing seeds with reduced levels of raffinose saccharides and phytic acid
US6013861A (en) Plants and processes for obtaining them
CA2069422C (en) A starch-derived, food-grade, insoluble bulking agent
US4777057A (en) Process for making a packaged dough for a baked confectionery
US6379968B1 (en) Transgenic plants or algae expressing an AGP enzyme coupled to a transit peptide
US6635804B2 (en) Nucleic acid molecules encoding soluble starch synthases from maize
AU690517B2 (en) Novel plants and processes for obtaining them
EP0874908B1 (en) Nucleic acid molecules from plants encoding enzymes which participate in the starch synthesis
JP4923171B2 (en) Wheat was modified activity of branching enzymes, and starch and starch containing products obtained therefrom
JP5283814B2 (en) Starch and starch-containing product Ssii activity decreased is barley and amylopectin content decreased
EP0987949B1 (en) Milled cereal by-product which is an additive for flour and dough
CA1308709C (en) Starch and products produced therefrom
KR101274849B1 (en) Rice and products thereof having starch with an increased proportion of amylose
EP1950303A1 (en) Genetically modified plants which synthesise a starch with low amylase content and higher swelling ability

Legal Events

Date Code Title Description
EEER Examination request
FZDE Dead