GB2586970A - Plants - Google Patents

Abstract

A method of producing Sorghum bicolor plants displaying day length neutrality that are propagatable from seed and which display at least one trait selected from i) ultra-earliness; ii) seeds being capable of sprouting and growing into mature plants at a latitude of up to 64 degrees North (N) in Europe; iii) display cold tolerance in the seed head at latitudes of up to 55 degrees North (N) in Europe; and iv) being able to mature under low light intensity. Further disclosed are sorghum plants that grow at latitudes from 30 degrees up to 64 degrees North and show substantial uniformity in inheritance of characteristics selected from: day length neutrality, high sugar level in the stem, cold tolerance in the seeds head, or increased levels of antioxidants in the leaves. Further disclosed are the seed and sorghum plants of CN7-8, W18, and HD6 having NCIMB deposit numbers 43458, 43459, and 43460, respectively.

Description

PLANTS

The present invention relates to crop plants and parts thereof, seeds, methods for the production of plants and seeds and uses therefor. In particular, the invention relates to Sorghum bicolor plants suitable for growing and cropping under European weather conditions at northern latitudes of from 300 North up to 64° North.

Crop plants of the species Sorghum bicolor are grown in tropical regions of Africa, Asia and South America for their seeds. Sorghum seeds are utilised in sorghum liquor production, typically for beer brewing processes, as a human food staple, and for animal fodder. In brewing processes, the seeds are used in producing sorghum wort, a liquor from which beer and other alcoholic liquors may be manufactured Seeds are also used to make sorghum meal (a kind of porridge), sorghum flour for baked products such as sorghum bread, sorghum pasta and other seed-based products for human consumption.

The grain, stems and leaves of sorghum plants grown in tropical regions are typically used as animal fodder (feed), the grains, leaves and stems of sorghum containing nutritive components. Leaf and stem plant material is also used as a primary resource material for the fencing and other industries. As such, sorghum plants grown in the tropics provide a useful and C4-carbon neutral source of primary materials for various industries.

Sorghum plants grown in sub-Saharan Africa are able to produce significant biomass and grain yield in poor environments, even when grown in semi-arid regions, and display distinct anatomical, morphological and physiological features that have enabled them to adapt and thrive in, for example, water limited environments. Sorghum has potential to become important as a mainstream crop in regions outside of the tropical zone.

Sorghum plants grow quickly in tropical areas and can be cropped up to two or three times a year, thus making it a potentially more desirable crop to grow than maize (also referred to as 'corn' in the art), a competitor staple product which requires up to 40% more water. In addition, it is thought that sorghum seed typically contains more antioxidants per unit volume of biomass than does maize and is thought to make for a healthy and nutritious staple for the human and/or animal diet.

While the agricultural world looks to constantly improve the availability of food and new plant products for use in manufacturing, sorghum has not been considered hitherto as a viable crop plant for growing in climes outside of tropical zones, in particular in northern and southern climes more than 10° either side of the equator. Sorghum plants grown in the tropics are short day plants (SD) which do not display day length neutrality, a necessary attribute for producing cold tolerant seed heads. Furthermore, sorghum plants are not generally grown at northern latitudes above about 10°N of the Equator or at southern latitudes, below about 10°S of the Equator.

Surprisingly, the inventor has, after many years, been able to produce sorghum plants of several different kinds which are capable of growing and maturing in northern climes above 10°N in Europe and in southern climes below 10°S, in for example, South America.

According to the present invention there is provided a method of producing sorghum plants displaying day length neutrality that are propagatable from seed and which display at least one trait selected from i) ultra-earliness; ii) seeds being capable of sprouting and growing into mature plants at latitudes from 50° North up to 64° North (N) in Europe; iii) display cold tolerance in the seed head at latitudes from 30° North up to 550 North (N) in Europe; and iv) being able to mature under low light intensity comprising: a) producing sorghum plants wherein each plant displays the trait of day length neutrality; b) selecting and crossing plants of step a) over at least four generations and selecting plant lines that reliably display at least the selectable trait of day length neutrality in Europe; c) selecting and crossing the plants obtained through step b) with other sorghum plants displaying at least one of the traits selected from i) ultra-earliness; ii) being capable of growing into mature plants at latitudes from 500 North up to 64° North (N) in Europe; iii) display cold tolerance in the seed head at latitudes from 30° North up to 55° North (N) in Europe; and iv) being able to mature under low light intensity; and d) producing sorghum plant populations that, when crossed, will reliably produce a population of plants that substantially uniformly display at least one of the traits i) to iv; e) harvesting the seeds of said crosses; f) sowing said seeds; and g) growing sorghum plants from the seeds of step f).

A Sorghum bicolor plant of the invention that displays 'ultra-earliness' is one which is capable of producing a fodder crop with a dry matter content above 30% from late August onwards, being cropped in Northern latitudes or Southern latitudes as defined herein. Ultra-earliness means that the sorghum plants are able to produce ripe seeds, preferably from 5 to 10 days earlier at northern latitudes, and also are 'from 30 to 35 days earlier' than conventional early sorghums which produce ripe seed at between 100 to 150 days, post sowing, at tropical latitudes, such as at the equator. Thus, sorghum plants displaying ultra-earliness are ones which are able to produce ripe seed in September from plants sown in May in northern climes and corresponding southern climes, including tropical climes, such as equatorial climes as defined herein. This is important for sorghum plants grown in northern and southern non-equatorial climes because it means that plants having the trait of ultra-earliness can be sown in a period where the risk of frost is low and seeds can be harvested before first frosts occur. Sorghum plants of the invention ('ultra-early' sorghum plants) that display ultra-earliness are typically able to produce ripe seed from 70 to 85 days after sowing, depending on latitude. This compares favourably with conventional sorghum plants displaying the trait of 'earliness' that are able to produce ripe seed from 100 to 150 days after sowing, again depending on latitude. Generally, conventional sorghum plants cannot be successfully grown and seed harvested therefrom in latitudes outside of 30reither side of the equator because they lack the ability to respond well to low irradiation levels of light. Such conventional sorghum plants that display 'earliness' ('early sorghum plants') are not able to mature efficiently and set seed quick enough for northern and southern climes as defined herein.

An internationally acceptable scale for recording the development stages of cereals, including sorghum, as they can be readily observed in the field was developed by Zadoks J.0 et al., 'A Decimal Code for the Growth Stages of Cereals', Weed Res. 14:415-21 (1974). Ultra-earliness of sorghum plants of the invention attracts a Decimal Code value of >90, preferably a Decimal Code value of 91.

A Sorghum bicolor plant displaying 'day length neutrality' is one which is able to flower independently of the day-length. Such plants are referred to in the art as day length neutral. They typically flower in a photoperiod which is neither long nor short. Conventional Sorghum bicolor plants grown in the tropics are not day length neutral. Sorghum bicolor plants in their native tropical habitat have short day (SD) photoperiodicity and are known as SD plants. Thus, conventional Sorghum bicolor plants in their native tropical habitat do not display the trait of day length neutrality.

For the purposes of the present invention 'sorghum plants', 'sorghum plant', 'Sorghum bicolor plant', Sorghum bicolor (L.) Moench and 'Sorghum bicolor plants' are all used interchangeably in the singular and/or plural, depending on context and refer to the same thing. Similarly, 'seed' and 'seeds' is used interchangeably in the singular and/or plural, depending on context and refer to the same thing.

In a preferment, there is provided a method of producing sorghum plants displaying day length neutrality that are propagatable from seed and which display at least the trait of cold tolerance in the seed head and are capable of producing seeds that sprout and grow into mature plants that set seed in northern latitudes of from about 30°N to about 55°N. Plants of this aspect of the invention are able to set seed in, for example, Sweden, Denmark and The Netherlands.

In a further preferment, there is provided a method of producing Sorghum plants displaying day length neutrality that are propagatable from seed and which display at least the trait of growing to full size but do not reliably set seed in latitudes of from 55°N up to 64° North (N) in Europe. Such plants can be promoted and used in the diet of domestic livestock and so can be used successfully as animal fodder in northern latitudes.

In another preferment, there is provided a method of producing sorghum plants displaying the trait of day length neutrality that are propagatable from seed and which display at least the further trait of earliness. 'Earliness' for the purposes of the invention means that the sorghum plants of the invention are capable of producing ripe seeds within 106 to 130 days from sowing, depending on latitude, sowing date and soil-type.

Cold tolerance in the seed head' means that Sorghum plants of the invention are able to reliably produce seed capable of germinating at a latitude of up to 550 North (N) in Europe. Furthermore, it has been found that sorghum plants displaying the trait of cold tolerance in the seed head in such northern climes also display the said trait in cooler southern climes, between the southern latitudes of from 30°S to 40°S, preferably from about 34°S to 40°S, such as 33°S, 34°S, 35°S, 36°S, 37°S, 38°S, 39°S, and 40°S and at any latitude therein between.

A Sorghum bicolor plant of the invention that displays the trait of 'being able to mature under low light intensity' is one which is capable of maturing in Northern latitudes or Southern latitudes as defined herein under a light intensity, measured as a downward thermal annual infrared measurement of an average value found between 5 to 9.5 kW-hr/m2/day and between 5,5 to 9,5 kW-hr/m2/day, preferably from 6,5 to 9,5 kW-hr/m2/day. This trait is not known to be present in conventional sorghum plants.

Plants of the invention, that is, plants being able to mature under low light intensity are able to grow and produce ripe seed in a temperature range of from about 12°C to about 20°C from sowing to harvest, preferably from 12°C to 20°C.

Europe' means countries in Europe such as Poland and also includes Russia, Belarus and The Ukraine. Preferably, Europe for the purposes of the present invention includes countries having weather patterns that are influenced by passing above water (seas). Sorghum plants of the invention typically grow and set seed in northern European latitudes of from about 35°N to about 55°N such as 35°N, 36°N, 37°N, 38°N, 39°N, 40°N, 41°N, 42°N, 43°N, 44°N, 45°N, 46°N, 47°N, 48°N, 49°N, 50°N, 51°N, 52°N, 53°N, 54°N and at any latitude therein between. Preferably, seed producing sorghum plants of the invention set seed within northern latitudes in the range of 40°N to 55°N, more preferably in the range of 44°N to 55°N, still more preferably in the range from 46°N to 53°N, and most preferably within the range of 48°N to 52°N. Plants produced according to the method of the invention and which set seed in northern climes such as at 51°N include the restorer lines W18 (grain ( chalky white), CN7-8 ( grain light red-brown) and I4D6 ( grain highly coloured dark red), deposits of which have been made at the NCIMB, Aberdeen, Scotland under NUMB Numbers 43459, 43458 and 43460, respectively.

Sorghum plants of the invention which do not set seed but which have healthy stems and leaves at northern latitudes as herein defined may be grown for non-seed related products such as animal feed, fencing materiel, and hardboard production from about 55°N to 64°N, such as 55°N, 56°N, 57°N, 58°N, 59°N, 60°N, 61°N, 62°N, 63°N and 64°N and at any latitude therein between. Examples of plants produced according to the method of the invention and which have healthy stems and leaves at latitudes of up to about 55°N to 64°N are plants W18, CN7-8, and HD6.

Naturally, the man skilled in the art will appreciate that many varieties of plants of the invention will be capable of either being grown on major land masses in northern and southern climes such as the United States of America, Russia, Belarus, Ukraine, Argentina, Chile, South Africa and China and/or will be capable of serving as parent plants for producing further varieties of Sorghum bicolor which may be bred for growing conditions found in the hinterland of such land masses. Thus, the man skilled in the art will appreciate that sorghum plants of the invention will be able to grow and display all the traits alluded to herein in locales outside of Northern Europe which display similar temperate climates thereto, such as areas in North America, for example in the current wheat belt, and in South America, for example Patagonia, China and South Africa. Sorghum plants grown at these latitudes typically possess medium to high brix content and can be promoted and used in the diet of domesticated livestock. 'Domesticated livestock' for the purposes of the invention means animals selected from mammals and birds that are bred for commercial purposes. Examples of domesticated livestock to which sorghum plants and/or seeds of the invention or parts thereof may serve as fodder include pigs, cattle, sheep, goats, horses, chickens, geese, ducks, racing pigeons and game birds such as pheasants, partridges, quails and the like.

In a further preferment there is provided Sorghum plants of the invention displaying day length neutrality that are propagatable from seed and which display at least the trait of growing to full size but do not reliably set seed in latitudes of from 41°S to 45°S in South America (Patagonia). Such plants can be used in the diet of domestic livestock and so can be grown for livestock fodder in southern latitudes.

In a further embodiment, the method of the invention provides sorghum plants capable of growing at latitudes of from 30° North up to 550 North that are: i) selected having at least a further trait selected from ultra-earliness, cold tolerance in the seed head, high sugar levels in the stem, high levels of antioxidants in the leaves, and high levels of antioxidants in the seed; and inbreeding the sorghum plants until the said further trait or traits of interest is/are stably inherited and wherein the inbred sorghum plants obtained are capable of being reliably reproduced in further generations at latitudes of from 30° North up to 55° North.

The number of inbreeding steps is typically between 4 and 10.

Typically, the parental populations of step c) show substantial uniformity of inheritance for one or more traits or characteristics selected from the group consisting of ultra-earliness, being able to mature under low light intensity, display day length neutrality, cold tolerance in the seed head, substantial sugar levels in the stem, and an ability to set seed at latitudes in Europe from 30° North up to 55° North.

In a further embodiment of the invention there is provided Fi sorghum plants produced by methods of the invention wherein said plants display the traits of ultra-earliness, day length neutrality and cold tolerance in the seed heads at latitudes from 30° North up to 55° North. Furthermore, such plants of the invention display the further traits of being capable of growing at latitudes between 55° North and 64° North and display at least the trait of high sugar levels in the stem.

In a further embodiment there is provided seed of sorghum plants of the invention.

In another embodiment of the invention there is provided a method of producing Fi hybrid sorghum plants from the plants obtained from the methods outlined hereinabove by: a) selecting and crossing a first parental line or clone with a second parental line or a clone that, when crossed, will reliably produce Fi plants that bear seeds; and wherein both parental lines or clones thereof show substantial uniformity in inheritance of characteristics selected from the group consisting of cold tolerant seed heads (e.g. temperatures around 12°C to 20°C), substantial sugar levels in the stem, ability to set seed in a wider range of latitudes than that of conventional (SD) sorghum plants; and b) harvesting the seed of said cross; wherein the germination percentage of the seed of b) is at least 60% and the germinated plants mature into sorghum plants that produce viable seeds at latitudes from 30° North up to 55° North.

Thus, there is further provided F1 hybrid sorghum seed produced according to the method outlined hereinabove, and plants grown from such hybrid sorghum seed. Furthermore, there is provided Sorghum plants that grow at a latitudes from 55° North up to 64° North which show substantial uniformity in inheritance of characteristics selected from the group consisting of ultra-earliness, day length neutrality, high sugar levels in the stem, and increased levels of antioxidants in the plant.

It will be apparent from the foregoing description that Sorghum plants (Sorghum bicolor L. Moench) are typically bred using self-pollination techniques. Using mechanisms employing male sterility (either genetic or cytoplasmic) and cross pollination breeding techniques may also be employed. Natural pollination occurs in sorghum when the anthers (male flowers) open and pollen falls onto receptive stigma (female flowers). Successful self-pollination in sorghum plants is very high (average 94%). Cross pollination may occur when wind or convection currents move pollen from the anthers of one plant to receptive stigma on another plant. Cross pollination is greatly enhanced using male sterility techniques which renders male flowers nonviable without affecting the female flowers. Successful pollination in the case of male sterile flowers requires cross pollination.

Sorghum is in the same family as maize and has a similar growth habit, but with more tillers and a more extensively branched root system. Sorghum is more drought tolerant and heat-tolerant than maize. Wild species of sorghum tend to grow to a height of 1.5 to 2 metres or more. However, in order to improve harvesting ability, dwarfing genes have been selected in cultivated varieties and hybrids such that most cultivated varieties and hybrids grow to between 60 and 120 cm tall.

As alluded to hereinabove, the development of sorghum hybrids requires the use of homozygous inbred lines, the crossing of these lines, and the evaluation of the crosses. Pedigree breeding methods may be used in preference to population breeding methods (as described above) to develop inbred lines how breeding populations. Breeding programs combine desirable traits from two or more inbred lines into breeding pools from which new inbred lines are developed by selling and selection of desired phenotypes. The new inbred lines may be crossed with other inbred lines and resultant hybrids from these crosses may be evaluated to determine commercial potential.

Pedigree breeding starts with the crossing of two genotypes, each of which may have one or more desirable characteristics that is lacking in the other or which complement the other. If the two original parents do not provide all of the desired characteristics, other sources can be included in the breeding population. In the pedigree method, superior plants are selfed and selected in successive generations. In the succeeding generations the heterozygous condition gives way to homogeneous lines as a result of self-pollination and selection. Typically, in the pedigree method of breeding five or more generations of selfing and selection is practiced. Fi to F2; F2 to F3; F3 to F4, F4 to F5, and so on.

Backcrossing can be used to improve an inbred line. Backcrossivag transfers a specific desirable trait from one inbred or source to an inbred that lacks that trait. This can be accomplished for example by first crossing a superior inbred (A) (recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate genes(s) for the trait in question. The progeny of this cross is then crossed back to the superior recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny will be heterozygous for loci controlling the characteristic being transferred but will be genetically similar to the superior parent for most or almost all other genes. The last backcross generation is typically selfed to give pure breeding progeny for the gene(s) being transferred.

Sorghum varieties are mainly self-pollinated; therefore, self-pollination of the parental varieties must be controlled to make hybrid development feasible. A pollination control system and effective transfer of pollen from one parent to the other offers improved plant breeding and an effective method for producing hybrid seed and plants.

Use of male sterility systems is an acknowledged means by which hybrid seed and plants of commercial use may be obtained. An example of a cytoplasmic male sterility method that may be employed in hybrid seed production is the milo or Ai cytoplasmic male sterility (CMS) system, developed via a cross between milo and lcafir cultivars, and is frequently used in hybrid sorghum production (Stephens J C & Holland P F, Cytoplasmic Male Sterility for Hybrid Sorghum Seed Production, Agron. J. 46:20-23 (1954)). Other CMS systems for sorghum include, but are not limited to, A2, isolated from IS 12662c (Schertz K F, Registration of AzTx 2753 and BT x 2753 Sorghum Gerrnplasm, Crop Sci. 17: 983 (1977)), A3, isolated from IS 1112c or converted Nilwa (Quinby.1 R, Interactions of Genes and Cytoplasms in Male-Sterility in Sorghums, Proc. 35th Corn Sorghum Res. Conf. Am. Seed Trade Assoc. Chicago, III., pp. 5-8 (1980)), Ar, isolated from IS 7920e (Worstell et al, Relationship among Male-Sterility Inducing Cytoplasms of Sorghum, Crop Sci. 24:186-189 (1984)).

In developing improved new sorghum hybrid varieties, breeders may use a CMS plant as the female parent. In using these plants, breeders may improve the efficiency of seed production and the quality of the F1 hybrids and so reduce breeding costs. When hybridization is conducted without using CMS plants, it is more difficult to obtain and isolate the desired traits in the progeny (F1 generation) because the parents are capable of undergoing both cross-pollination and self-pollination. If one of the parents is a CMS plant that is incapable of producing pollen, only cross pollination will occur. By eliminating the pollen of one parental variety in a cross, a plant breeder is assured of obtaining hybrid seed of uniform quality, provided that the parents are of uniform quality and the breeder conducts a single cross.

Thus, in one technique the production of Ft hybrid plants may include crossing a CMS female parent with a pollen-producing male parent To reproduce effectively, however, the male parent of the Fr hybrid must have a fertility restorer gene (RI' gene). The presence of an RI gene means that the Fi generation will not be completely or partially sterile, so that either self-pollination or CTOSS pollination may occur. Self-pollination of the Fi generation to produce several subsequent generations ensures that a desired trait is heritable and stable and that a new variety has been isolated.

Promising advanced breeding lines commonly are tested and compared to appropriate standards in environments representative of the commercial target area(s). The best lines are candidates for new commercial lines; and those still deficient in a few traits may be used as parents to produce new populations for further selection.

A hybrid sorghum variety is the cross of two inbred lines. The hybrid progeny of the first generation is designated F1. In the development of hybrids only the Fi hybrid plants are sought. The Fi hybrid is more vigorous than its inbred parents. Thus, hybrid vigour, or heterosis, may be observed in many ways, such as in increased vegetative growth and increased yield.

The development of a hybrid sorghum variety typically involves five steps: (1) the formation of "restorer" and "non-restorer" germplasm pools; (2) the selection of superior plants from various "restorer" and "non-restorer" germplasm pools; (3) the selfmg of the superior plants for several generations to produce a series of inbred lines, which although different from each other, each breed true and are highly uniform; (4) the conversion of inbred lines classified as non-restorers to cytoplasmic male sterile (CMS) forms, and (5) crossing the selected cytoplacmic male sterile (CMS) inbred lines with selected fertile inbred lines (restorer lines) to produce the hybrid progeny (FO.

Since sorghum is normally a self-pollinated plant and because both male and female flowers are in the same panicle, large numbers of hybrid seed can only be produced by using cytoplasmic male sterile (CMS) inbred plants. Flowers of the CMS inbred are fertilized with pollen from a male fertile inbred carrying genes which restore male fertility in the hybrid (Ft) plants. Once the inbred plants that produce the best hybrid have been identified, the hybrid seed can be reproduced indefinitely as long as the homogeneity of the inbred parents is maintained.

A single cross hybrid is produced when two inbred lines are crossed to produce the F1 progeny. Hybrid vigour exhibited by F; hybrids is generally lost in the next generation (F2). Consequently, seed from hybrid varieties is not used for planting stock.

Hybrid sorghum can be produced using wind to transfer the pollen. Alternating strips of cytoplasmic male sterile inbred (female) and male fertile inbred (male) lines are planted in the same field. Wind transfers the pollen shed by the male inbred to receptive stigma on the female. Providing that there is sufficient isolation from sources of foreign sorghum pollen, the stigma of the male sterile inbred line (female) will be fertilized only with pollen from the male fertile inbred line (male). The resulting seed, born on the male sterile (female) plants is therefore hybrid and will form hybrid plants that have full fertility restored.

In addition to phenotypic observations, a plant can also be described or identified by its genotype. The genotype of plants, for example those deposited with the NCIMB as herein described can be characterized through a genetic marker profile. Genetic marker profiles can be obtained by techniques such as Restriction Fragment Length Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats (SSRs) which are also referred to as Microsatellites, and Single Nucleotide Polymorphisms (SNPs).

Particular markers used for these purposes may include any type of marker and marker profile which provides a means of distinguishing varieties. A genetic marker profile can be used, for example, to identify plants of the same variety or related varieties or to determine or validate a pedigree. In addition to being used for identification of Fi sorghum hybrids, their inbred parents, such as those deposited with the NCIMB as described herein and their plant parts, the genetic marker profile is also useful in developing a locus conversion of any Fi hybrid plant that may be the result of crosses of parent lines which display substantial uniformity of desired traits as described herein.

Methods of isolating nucleic acids from sorghum plants and methods for performing genetic marker profiles using SNP and SSR polymorphisms are well known in the art. SNPs are genetic markers based on a polymorphism in a single nucleotide. A marker system based on SNPs can be highly informative in linkage analysis relative to other marker systems in that multiple alleles may be present.

A method comprising isolating nucleic acids, such as DNA, from a plant, a plant part, plant cell or a seed of the sorghum plants disclosed herein is provided. The method can include mechanical, electrical and/or chemical disruption of the plant, plant part, plant cell or seed, contacting the disrupted plant, plant part, plant cell or seed with a buffer or solvent, to produce a solution or suspension comprising nucleic acids, optionally contacting the nucleic acids with a precipitating agent to precipitate the nucleic acids, optionally extracting the nucleic acids, and optionally separating the nucleic acids such as by centrifugation or by binding to beads or a column, with subsequent elution, or a combination thereof. If DNA is being isolated, an RNase can be included in one or more of the method steps. The nucleic acids isolated can comprise all or substantially all of the genomic DNA sequence, all or substantially all of the chromosomal DNA sequence or all or substantially all of the coding sequences (cDNA) of the plant, plant part, or plant cell from which they were isolated. The amount and type of nucleic acids isolated may be sufficient to permit whole genome sequencing of the plant from which they were isolated or chromosomal marker analysis of the plant from which they were isolated.

The methods can be used to produce nucleic acids from the plant, plant part, seed or cell, which nucleic acids can be, for example, analyzed to produce data. The data can be recorded. The nucleic acids from the disrupted cell, the disrupted plant, plant part, plant cell or seed or the nucleic acids following isolation or separation can be contacted with primers and nucleotide bases, and/or a polymerase to facilitate PCR sequencing or marker analysis of the nucleic acids. In some examples, the nucleic acids produced can be sequenced or contacted with markers to produce a genetic profile, a molecular profile, a marker profile, a haplotype, or any combination thereof. In some examples, the genetic profile or nucleotide sequence is recorded on a computer readable medium. In other examples, the methods may further comprise using the nucleic acids produced from plants, plant parts, plant cells or seeds in a plant breeding program, for example in making crosses, selection and/or advancement decisions in a breeding program. Crossing includes any type of plant breeding crossing method, including but not limited to crosses to produce hybrids, outcrossing, selfing, backcrossing, locus conversion, introgression and the like. Desirable genotypes and or marker profiles, optionally associated with a trait of interest, may be identified by one or more methodologies. In some examples one or more markers are used, including but not limited to AFLPs, RFLPs, ASH, SSRs, SNPs, indels, padlock probes, molecular inversion probes, microarrays, sequencing, and the like. In some methods, a target nucleic acid is amplified prior to hybridization with a probe. In other cases, the target nucleic acid is not amplified prior to hybridization, such as methods using molecular inversion probes. In some examples, the genotype related to a specific trait is monitored, while in other examples, a genome-wide evaluation including but not limited to one or more of marker panels, library screens, association studies, microarrays, gene chips, expression studies, or sequencing such as wholegenome resequencing and genotyping-by-sequencing (GBS) may be used. In certain instances, no target-specific probe is needed, for example by using sequencing technologies, including but not limited to next-generation sequencing methods (see, for example, Metzker (2010) Nat Rev Genet 11:31-46; and, Egan et al. (2012) Am J Bot 99:175-185) such as sequencing by synthesis (e.g., Roche 454 pyrosequencing, Illumina Genome Analyzer, and Ion Torrent PGM or Proton systems), sequencing by ligation (e.g., SOLiD from Applied Biosystems, and Polnator system from Azco Biotech), and single molecule sequencing (SMS or third-generation sequencing) which eliminate template amplification (e.g., Helicos system, and PacHio RS system from Pacific BioSciences). Further technologies include optical sequencing systems (e.g., Starlight from Life Technologies), and nanopore sequencing (e.g., GridION from Oxford Nanopore Technologies). Each of these may be coupled with one or more enrichment strategies for organellar or nuclear genomes in order to reduce the complexity of the genome under investigation via PCR, hybridization, restriction enzyme, and expression methods. In some examples, no reference genome sequence is needed in order to complete the analysis. The plant lines deposited with the NCIMB as described herein and their plant parts and Fi hybrids derived therefrom can be identified through a molecular marker profile. Such plant parts may be either diploid or haploid. The plant part includes at least one cell of the plant from which it was obtained, such as a diploid cell, a haploid cell or a somatic cell. Also provided are plants and plant parts substantially benefiting from the use of such NCIMB deposited plants in their development, such as F1 hybrid plants comprising a locus conversion.

Locus Conversions ofFi Hybrid plants derived from parent plants encompassed by the invention may provide new base genetic lines into which new loci or traits may be introduced. Direct transformation and backcrossing represent two methods that can be used to accomplish such an introgression. The term 'locus conversion' is used to designate the product of such an introgression.

To select and develop a desirable, typically a superior hybrid, it is necessary to identify and select genetically unique individuals that occur in a segregating population. The segregating population is the result of a combination of crossover events plus the independent assortment of specific combinations of alleles at many gene loci that results in specific and unique genotypes. Locus conversions can be used to add or modify one or a few traits of such a line such as yield, disease resistance, pest resistance and plant performance in varying or extreme weather conditions.

Backcrossing can be used to improve inbred varieties and a hybrid variety which is made using those inbred lines. Backcrossing can be used to transfer a specific desirable trait from one plant line, the donor parent, to an inbred line referred to as a 'recurrent parent' which has overall good agronomic characteristics yet that lacks the desirable trait This transfer of the desirable trait into an inbred with overall good agronomic characteristics can be accomplished by first crossing a recurrent parent to a donor parent (non-recurrent parent). The progeny of this cross is then mated back to the recurrent parent followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent.

Traits may be used by those of ordinary skill in the art to characterize progeny. Traits are commonly evaluated at a significance level, such as a 1%, 5% or 10% significance level, when measured in plants grown in the same environmental conditions. For example, a locus conversion of an Ft hybrid plant may be characterized as having essentially the same phenotypic traits as that plant. The traits used for comparison may be those traits as defined and described herein. Molecular markers can also be used during the breeding process for the selection of qualitative traits. For example, markers can be used to select plants that contain the alleles of interest during a backcrossing breeding program. The markers can also be used to select for the genome of the recurrent parent and against the genome of the donor parent. Using this procedure can minimize the amount of genome from the donor parent that remains in the selected plants.

A locus conversion of an Ft hybrid should retain the genetic integrity of that Ft hybrid. For example, a locus conversion of an Ft hybrid can be developed when DNA sequences are introduced through backcrossing (Hanauer et al., 1988), with a parent of the Ft hybrid utilized as the recurrent parent Both naturally occurring and transgenic DNA sequences may be introduced through backcrossing techniques. A backcross conversion may produce a plant with a locus conversion in at least one or more backcrosses, including at least 2 crosses, at least 3 crosses, at least 4 crosses, at least 5 crosses and so on. Molecular marker assisted breeding or selection may be utilized to reduce the number of backcrosses necessary to achieve the backcross conversion. For example, see Openshaw, S. J. et al., Marker-assisted Selection in Backcross Breeding. In: Proceedings Symposium of the Analysis of Molecular Data, August 1994, Crop Science Society of America, Corvallis, Oreg., where it is demonstrated that a backcross conversion can be made in as few as two backcrosses. A locus conversion of an F1 hybrid can be determined through the use of a molecular profile. A locus conversion of an Fi hybrid would have 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the molecular markers, or molecular profile, of the F1 hybrid. Examples of molecular markers that could be used to determine the molecular profile include Restriction Fragment Length Polymorphisms (RFLP), Polymerase Chain Reaction (PCR) analysis, and Simple Sequence Repeats (SSR), and Single Nucleotide Polymorphisms (SNPs).

In addition, genetic modification and transformation of sorghum plants such as Fi hybrid plants and/or parent lines thereof may include the use of transgenes, genetic or gene editing or modification and transformation methods used to facilitate engineering of the genome of*plants to introduce and express heterologous genetic elements, such as foreign genetic elements, additional copies of endogenous elements, or modified versions of native or endogenous genetic elements in order to alter at least one trait of a plant in a specific manner. Any sequences, such as DNA, whether from a different species or from the same species, which have been stably inserted into a genome using transformation are referred to herein collectively as "transgenes" and/or "transgenic events". Transgenes can be moved from one genome to another using breeding techniques which may include crossing, backcrossing or double haploid production. In some embodiments, a transformed variant of an Fi hybrid or parent line thereof may comprise at least one transgene or genetic modification but could contain at least 1,2, 3,4, 5, 6, 7, 8, 9, 10 or more transgenes and/or genetic modifications, depending on end purpose. The man skilled in the art will appreciate that the number of modifications that may be required to alter the genotype and/or phenotype of a plant will be dictated by a desired target genotype and/or phenotype for a given end purpose.

Numerous methods for plant transformation have been developed, including biological and physical plant transformation protocols. See, for example, Miki et al., "Procedures for Introducing Foreign DNA into Plants" in Methods in Plant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages 67-88 and Armstrong, "The First Decade of Maize Transformation: A Review and Future Perspective" (Maydica 44:101-109, 1999). In addition, expression vectors and in vitro culture methods for plant cell or tissue transformation and regeneration of plants are available. See, for example, Gruber et al., "Vectors for Plant Transformation" in Methods in Plant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J. E. Pus. (CRC Press, Inc., Boca Raton, 1993) pages 89-119.

In general, methods to transform, modify, edit or alter plant endogenous genomic DNA include altering the plant native DNA sequence or a pre-existing transgenic sequence including regulatory elements, coding and non-coding sequences. These methods can be used, for example, to target nucleic acids to pre-engineered target iccugnition sequences in the genome. Such pre-engineered target sequences may be introduced by genome editing or modification. As an example, a genetically modified plant line, such as a plant variety is generated using "custom" or engineered endonucleases such as meganucleases produced to modify plant genomes (see e.g., WO 2009/114321; Gao et at (2010) Plant Journal 1:176187). Another site-directed engineering method is through the use of zinc finger domain recognition coupled with the restriction properties of restriction enzymes. See e.g., limey, et al., (2010) Nat Rev Genet. 11(9):636-46; Shulda, et al., (2009) Nature 459 (7245):437-41. A transcription activator-like ('ML) effector-DNA modifying enzyme (TALE or TALEN) may also be used to engineer changes in the plant genome. See e.g., US20110145940, Cennak et al., (2011) Nucleic Acids Res. 39(12) and Boch et al., (2009), Science 326(5959): 1509-12. Site-specific modification of plant genomes may also be performed using the bacterial type II CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CR1SPRassociated) system. See e.g., Belhaj et al., (2013), Plant Methods 9: 39; The Cas9/guide RNA-based system allows targeted cleavage of genomic DNA guided by a customizable small noncoding RNA in plants (see e.g., WO 2015026883A1).

Plant transformation methods may involve the construction of an expression vector. Such a vector comprises a DNA sequence that contains a gene under the control of or operatively linked to a regulatory element, for example a promoter. The vector may contain one or more genes and one or more regulatory elements.

A transgenic event which has been engineered into a particular sorghum plant using transformation techniques, could be moved into another line using traditional breeding techniques that are well known to the man skilled in the art of plant breeding. For example, a backcrossing approach could be used to move a transgene from a transformed sorghum plant to an elite inbred line and the resulting progeny would comprise a transgene. Similarly, if an inbred line is used for the transformation then the transgenic plants could be crossed to a different line in order to produce a transgenic hybrid sorghum plant. As used herein, "crossing" can refer to a simple X by Y cross, or the process of backcrossing, depending on the context. Various genetic elements can be introduced into the plant genome using transformation. These elements include but are not limited to genes; coding sequences; inducible, constitutive, and tissue specific promoters; enhancing sequences; and signal and targeting sequences. For example, see, U.S. Pat No. 6,118,055.

The man skilled in the art knows that where a transgene for a foreign or exotic protein may be introduced into parent lines and Fl hybrids of the present invention, that protein may be produced in commercial quantities therein. Thus, techniques for the selection and propagation of transformed plants, which are well understood in the art, may yield a plurality of transgenic plants which are harvested in a conventional manner, and any introduced foreign protein then can be extracted from a tissue of interest or from total biomass. Protein extraction from plant biomass can be accomplished by known methods as discussed, for example, by Ileney and On, (1981) AnaL Biochem. 114:92-96.

A genetic map can be generated, primarily via conventional Restriction Fragment Length Polymorphisms (RFLP), Polymerase Chain Reaction (PCR) analysis, and Simple Sequence Repeats (SSR), and Single Nucleotide Polymoiphisms (SNPs), which identifies the approximate chromosomal location of the integrated DNA molecule coding for the foreign protein. Exemplary methods may be found in, for example, Glick and Thompson, METHODS IN PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 269-284 (CRC Press, Boca Raton, 1993). Map information concerning chromosomal location is useful for proprietary protection of a subject transgenic plant. If unauthorized propagation is undertaken and crosses made with other germplasm, the map of the integration region can be compared to similar maps for suspect plants, to determine if the latter have a common parentage with the subject plant. Map comparisons would involve hybridizations, RFLP, PCR, SSR, SNP, and sequencing, all of which are conventional techniques.

Thus, sorghum plants of the present invention may be genetically engineered to express various phenotypes of agronomic interest. Some exemplary genes of interest may be selected from the following non-exhaustive list: 1. Genes that Create a Site for Site Specific DNA Integration.

This includes the introduction of FRT sites that may be used in the FLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system. For example, see, Lyznik, et al., (2003) "Site-Specific Recombination for Genetic Engineering in Plants", Plant Cell Rep 21:925-932 and WO 99/25821, which are hereby incorporated by reference. Other systems that may be used include the Gin recombinase of phage Mu (Maeser, et al., 1991), the Pin recombinase of E. coil (Enomoto, et al., 1983), and the R/RS system of the pSR1 plasmid (Araki, et al., 1992).

2. Genes that affect abiotic stress resistance (including but not limited to flowering, panicle/glume and seed development, enhancement of nitrogen utilization efficiency, altered nitrogen responsiveness, drought resistance or tolerance, cold resistance or tolerance, and salt resistance or tolerance) and increased yield under stress. For example, see, WO 00/73475 where water use efficiency is altered through alteration of malate; U.S. Pat. Nos. 5,892,009, 5,965,705, 5,929,305, 5,891,859, 6,417,428, 6,664,446, 6,706,866, 6,717,034, 6,801,104, W02000060089, W02001026459, W02001035725, W02001034726, W02001035727, W02001036444, W02001036597, W02001036598, W02002015675, W02002017430, W02002077185, W02002079403, W02003013227, W02003013228, W02003014327, W02004031349, W02004076638, W09809521 and W09938977 describing genes, including CBF genes and transcription factors effective in mitigating the negative effects of freezing, high salinity, and drought on plants, as well as conferring other positive effects on plant phenotype; US Patent Application Publication Number 2004/0148654 and W001/36596 where abscisic acid is altered in plants resulting in improved plant phenotype such as increased yield and/or increased tolerance to abiotic stress; W02000/006341, W004/090143, U.S. patent application Ser. Nos. 10/817,483 and 09/545,334 where cytokinin expression is modified resulting in plants with increased stress tolerance, such as drought tolerance, and/or increased yield. Also see W00202776, W003052063, JP2002281975, U.S. Pat. No. 6,084,153, W00164898, U.S. Pat. Nos. 6,177,275 and 6,107,547 (enhancement of nitrogen utilization and altered nitrogen responsiveness). For ethylene alteration, see, US Patent Application Publication Numbers 2004/0128719, 2003/0166197 and W0200032761. For plant transcription factors or transcriptional regulators of abiotic stress, see e.g., US Patent Application Publication Number 2004/0098764 or US Patent Application Publication Number 2004/0078852.

Other genes and transcription factors that affect plant growth and agronomic traits such as yield, flowering, plant growth and/or plant structure, can be introduced or introgressed into plants, see, e.g., W097/49811 (LHY), W098/56918 (ESD4), W097/10339 and U.S. Pat. No. 6,573,430 (TFL), U.S. Pat. No. 6,713,663 (F1'), W096/14414 (CON), W096/38560, W001/21822 (VRN1), W000/44918 (VRN2), W099/49064 (GI), W000/46358 (FM), W097/29123, U.S. Pat. Nos. 6,794,560,6,307,126 (GAI), W099/09174 (D8 and Rht), and W02004076638 and W02004031349 (transcription factors).

3. Transgenes that Confer or Contribute to an Altered Grain Characteristic, including: A. Altered phosphorus content, for example, by the (1) Introduction of a phytase-encoding gene would enhance breakdown of phytate, adding more free phosphate to the transformed plant. For example, see, Van Hartingsveldt, et al., Gene 127:87 (1993), for a disclosure of the nucleotide sequence of an Aspergillus!tiger phytase gene. (2) Up-regulation of a gene that reduces phytate content. For example, this could be accomplished, by cloning and then reintroducing DNA associated with one or more of the alleles, such as the LPA alleles, identified in mutants characterized by low levels of phytic acid, such as in Raboy, et al. (1990).

B. Altered fatty acids, for example, by down-regulation of stearoyl-ACP desaturase to increase stearic acid content of the plant. See ICnult2on, et al., Proc. Natl. Acad. Sci. USA 8%2624 (1992).

C. Altered carbohydrates effected, for example, by altering a gene for an enzyme that affects the branching pattern of starch, a gene altering thioredoxin. (See, U.S. Pat. No. 6,531,648). See, Shiroza, et al., (1988) J. Bacteriol 170:810 (nucleotide sequence of Streptococcus mutans fructosyltransferase gene), Steinmetz, et al, (1985) Mol. Gen. Genet. 200:220 (nucleotide sequence of Bacillus subtilis levansucrase gene), Pen, et al., (1992) Bio/Technology 10:292 (production of transgenic plants that express Bacillus licheniformis alpha-amylase), Elliot, et al., (1993) Plant Molec Biol 21:515 (nucleotide sequences of tomato invertase genes), Sogaard, et al., (1993) J. Biol. Chem. 268:22480 (site-directed mutagenesis of barley alpha-amylase gene) and Fisher, et al., (1993) Plant Physiol 102:1045 (maize endosperm starch branching enzyme II), WO 99/10498 (improved digestibility and/or starch extraction through modification of UDP-D-xylose 4-epimerase, Fragile 1 and 2, Refl, HCHL, C4H), U.S. Pat. No. 6,232,529 (method of producing high oil seed by modification of starch levels (AGP)). The fatty acid modification genes mentioned above may also be used to affect starch content and/or composition through the interrelationship of the starch and oil pathways.

I). Altered antioxidant content or composition, such as alteration of tocopherol or tocotrienols. For example, see, U.S. Pat. No. 6,787,683, US Patent Application Publication Number 2004/0034886 and WO 00/68393 involving the manipulation of antioxidant levels through alteration of a phytIprenyl transferase (ppt), WO 03/082899 through alteration of a homogentisate geranyl geranyl transfera.se (hggt).

E. Altered essential seed amino acids. For example, see, U.S. Pat. No. 6,127,600 (method of increasing accumulation of essential amino acids in seeds), U.S. Pat. No. 6,080,913 (binary methods of increasing accumulation of essential amino acids in seeds), U.S. Pat. No. 5,990,389 (high lysine), W099/40209 (alteration of amino acid compositions in seeds), W099/29882 (methods for altering amino acid content of proteins), U.S. Pat. No. 5,850,016 (alteration of amino acid compositions in seeds), W098/20133 (proteins with enhanced levels of essential amino acids), U.S. Pat. No. 5,885,802 (high methionine), U.S. Pat. No. 5,885,801 (high threonine), U.S. Pat. No. 6,664,445 (plant amino acid biosynthetic enzymes), U.S. Pat. No. 6,459,019 (increased lysine and threonine), U.S. Pat. No. 6,441,274 (plant tryptophan synthase beta subunit), U.S. Pat. No. 6,346,403 (methionine metabolic enzymes), U.S. Pat. No. 5,939,599 (high sulfur), U.S. Pat. No. 5,912,414 (increased methionine), W098/56935 (plant amino acid biosynthetic enzymes), W098/45458 (engineered seed protein having higher percentage of essential amino acids), W098/42831 (increased lysine), U.S. Pat. No. 5,633,436 (increasing sulfur amino acid content), U.S. Pat. No. 5,559,223 (synthetic storage proteins with defined structure containing programmable levels of essential amino acids for improvement of the nutritional value of plants), W096/01905 (increased threonine), W095/15392 (increased lysine), US Patent Application Publication Number 2003/0163838, US Patent Application Publication Number 2003/0150014, US Patent Application Publication Number 2004/0068767, U.S. Pat. No. 6,803,498, W001/79516, and W000/09706 (Ces A: cellulose synthase), U.S. Pat. No. 6,194,638 (hemicellulose), U.S. Pat. No. 6,399.859 and US Patent Application Publication Number 2004/0025203 (UDPGdH), U.S. Pat No. 6,194,638 (RGP).

4. Genes that Confer Male Sterility There are several methods of conferring genetic male sterility available, such as multiple mutant genes at separate locations within the genome that confer male sterility, as disclosed in U.S. Pat. Nos. 4,654,465 and 4,727,219 to Brar, et al., and chromosomal translocations as described by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. In addition to these methods, Albertsen, et al., U.S. Pat. No. 5,432,068, describes a system of nuclear male sterility which includes: identifying a gene which is critical to male fertility; silencing this native gene which is critical to male fertility; removing the native promoter from the essential male fertility gene and replacing it with an inducible promoter; inserting this genetically engineered gene back into the plant; and thus creating a plant that is male sterile because the inducible promoter is not "on" resulting in the male fertility gene not being transcribed. Fertility is restored by inducing, or turning "on," the promoter, which in turn allows the gene that confers male fertility to be transcribed.

A. A dominant nuclear gene, Ms(tc) controlling male sterility. See, Elkortin, L. A., Theor. App!. Genet. (2005) 111(7): 1377-1384.

B. A tapetum-specific gene, RTS, a sorghum anther-specific gene is required for male fertility and its promoter sequence directs tissue-specific gene expression in different plant species. Luo, Hong, et al., Plant Molecular Biology., 62(3): 397-408(12) (2006). Introduction of a deacetylase gene under the control of a tapettun-specific promoter and with the application of the chemical N-Ac-PPT. See International Publication No. WO 01/29237.

C. Introduction of various stamen-specific promoters. Anther-specific promoters which are of particular utility in the production of transgenic male-sterile monocots and plants for restoring their fertility. See, U.S. Pat. No. 5,639,948. See also, International Publication Nos. WO 92/13956 and WO 92/13957.

D. Introduction of the bamase and the barstar genes. See, Paul, et al., Plant Mol. Biol., 19:611-622 (1992).

For additional examples of nuclear male and female sterility systems and genes, see also, U.S. Pat. Nos. 5,859,341, 6,297,426,5,478,369, 5,824,524, 5,850,014, and 6,265,640. See also, Hanson, Maureen R., et al., "Interactions of Mitochondrial and Nuclear Genes That Affect Male Gametophyte Development," Plant Cell., 16:5154-5169 (2004), all of which are hereby incorporated by reference.

A. Modification of RNA editing within mitochondria' open reading frames. See, Pring, D. R., et al, Curt Genet. (1998) 33(6): 429-436; Pring, D. R., et aL, J. Hered. (1999) 90(3): 386393; Pring, D. R., et al., Curr. Genet. (2001) 39(5-6): 371-376; and Hedgcoth, C., et al., CUIT. Genet. (2002) 41(5): 357-365.

B. Cytoplasmic male sterility (CMS) from mutations at atp6 codons. See, Kempken, F., FEBS. Lett. (1998): 441(2): 159-160.

C. Inducing male sterility through heat shock. See, Wang, L., Yi Chuan Xue Bao. (2000) 27(9): 834-838. D. Inducing male sterility through treatment of streptomycin on sorghum callus cultures. See, Elkonin, L. A., et al., Genetica (2008) 44(5): 663-673.

5. Transgenes that Confer Tolerance to an Herbicide. A non-exhaustive list of transgenes which confer tolerance to herbicides follows: (A) A herbicide that inhibits the growing point or meristem, such as an imidazolinone or a sulfonylurea. Exemplary genes in this category code for mutant acetolactate synthase (ALS) and acetohydroxyacid synthase (AHAS) enzyme as described, for example, in U.S. Pat. Nos. 5,605,011; 5,013,659; 5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107; 5,928,937; and 5,378,824; US Patent Publication No. 20070214515, and international publication WO 96/33270.

(B) Glyphosate (tolerance imparted by mutant 5-enolpyruv1-3-phosphikimate synthase (EPSP) and aroA genes, respectively) and other phosphono compounds such as glufosinate (phosphinothricin acetyl transferase (PAT) and Streptomyces hygroscopicus phosphinothricin acetyl transferase (bar) genes), and pyridinoxy or phenoxy proprionic acids and cyclohexones (ACCase inhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835, which discloses the nucleotide sequence of a form of EPSPS which can confer glyphosate tolerance. U.S. Pat. No. 5,627,061 also describes genes encoding EPSPS enzymes. See also U.S. Pat. Not 6,566,587; 6,338,961; 6,248,876 Bl; 6,040,497; 5,804,425; 5,633,435; 5,145,783; 4,971,908; 5,312,910; 5,188,642; 4,940,835; 5,866,775; 6,225,11481; 6,130,366; 5,310,667; 4,535,060; 4,769,061; 5,633,448; 5,510,471; Re. 36,449; RE 37,287 E; and 5,491,288; and international publications EP1173580; WO 01/66704; EP1173581 and EP1173582.

Glyphosate tolerance is also imparted to plants that express a gene that encodes a glyphosate oxido-reductase enzyme as described more fully in U.S. Pat. Nos. 5,776,760 and 5,463,175. In addition, glyphosate tolerance can be imparted to plants by the over expression of genes encoding glyphosate N-acetyltransferase. See, for example, US2004/0082770; US2005/0246798; and US2008/0234130. A DNA molecule encoding a mutant aroA gene can be obtained under ATCC accession No. 39256, and the nucleotide sequence of the mutant gene is disclosed in U.S. Pat. No. 4,769,061. European Patent Application No. 0 333 033 and U.S. Pat. No. 4,975,374 disclose nucleotide sequences of glutamine synthetase genes which confer tolerance to herbicides such as L-phosphinothricin. The nucleotide sequence of a phosphinothricin-acetyl-transferase gene is provided in European Patent Nos. 0 242 246 and 0 242 236. See also, U.S. Pat. Nos. 5,969,213; 5,489,520; 5,550,318; 5,874,265; 5,919,675; 5,561,236; 5,648,477; 5,646,024; 6,177,616 Bl; and 5,879,903. Exemplary genes conferring resistance to phenoxy propionic acids, cyclohexanediones and eyclohexones, such as sethoxydim and haloxyfop, are the Accl -SI, Accl -S2 and Accl -S3 genes described by Marshall et al., Theor. Appl. Genet. 83: 435 (1992).

(C) A herbicide that inhibits photosynthesis, such as a triazine (psbA and gs+ genes) and a benzonitrile (nitrilase gene) such as bromoxynil. Przibilla et al., Plant Cell 3: 169 (1991), describe the transformation of Chlamydomonas with plasmids encoding mutant psbA genes. Nucleotide sequences for nitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, and DNA molecules containing these genes are available under ATCC Accession Nos. 53435, 67441 and 67442. Cloning and expression of DNA coding for a glutathione 5-transferase is described by Hayes et al., Biochem. J. 285: 173 (1992).

(D) Other genes that confer tolerance to herbicides include: a gene encoding a chimeric protein of rat cytochrome P4507A1 and yeast NADPH-cytochrome P450 wddoreductase (Shiota et al. (1994) Plant Physiol 106:17), genes for glutathione reductase and superoxide dismutase (Aono et al. (1995) Plant Cell Physiol 36:1687, and genes for various phosphotransferases (Datta etal. (1992) Plant Mol Biol 20:619).

(E) An herbicide that inhibits protoporphyrinogen oxidase (protox or PPO) is necessary for the production of chlorophyll, which is necessary for all plant survival. The protox enzyme serves as the target for a variety of herbicidal compounds. PPO-inhibitor herbicides can inhibit growth of all the different species of plants present, causing their total destruction. The development of plants containing altered protox activity which are tolerant to these herbicides are described, for example, in U.S. Pat. Nos. 6,288,306 Bl; 6,282,837 Bl; and 5,767,373; and international patent publication WO 01/12825.

(F) Dicamba (3,6-dichloro-2-methoxybenzoic acid) is an organochloride derivative of benzoic acid which functions by increasing plant growth rate such that the plant dies.

6. Transgenes that Confer Resistance to Insects or Disease and that Encode, for Example: (A) Plant disease resistance genes. Martin et al., Science 262: 1432 (1993) (tomato Pto gene for resistance to Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinos et al., Cell 78: 1089 (1994) (Arabidopsis RSP2 gene for resistance to Pseudomonas syringae), McDowell and Woffenden, (2003) Trends Biotechnol. 21(4): 178-83 and Toyoda et al., (2002) Transgenic Res. 11(6):567-82. A plant resistant to a disease is one that is more resistant to a pathogen as compared to the wild type plant.

(B) A Bacillus thuringiensis protein, a derivative thereof or a synthetic polypeptide modelled thereon. See, for example, Geiser et al., Gene 48: 109 (1986), who disclose the cloning and nucleotide sequence of a Bt delta-endotoxin gene. Moreover, DNA molecules encoding deltaendotoxin genes can be purchased from American Type Culture Collection (Rockville, Md.), for example, under ATCC Accession Nos. 40098, 67136, 31995 and 31998. Other non-limiting examples of Bacillus thuringiensis transgenes being genetically engineered are given in the following patents and patent applications: U.S. Pat. Nos. 5,188,960; 5,689,052; 5,880,275; 5,986,177; 7,105332; 7,208,474; WO 91/14778; WO 99/31248; WO 01/12731; WO 99/24581; WO 97/40162 and U.S. application Ser. Nos. 10/032,717; 10/414,637; 11/018,615; 11/404,297; 11/404,638; 11/471,878; 11/780,501; 11/780,511; 11/780,503; 11/953,648; and Ser. No. 11/957,893.

(C) An insect-specific hormone or pheromone such as an ecdysteroid and juvenile hormone, a variant thereof, a mimetic based thereon, or an antagonist or agonist thereof. See, for example, the disclosure by Hammock et al., Nature 344: 458 (1990), of baculovirus expression of cloned juvenile hormone esterase, an inactivator of juvenile hormone.

(D) An insect-specific peptide which, upon expression, disrupts the physiology of the affected pest. For example, see the disclosures of Regan, J. Biol. Chem. 269: 9 (1994) (expression cloning yields DNA coding for insect diuretic hormone receptor); Pratt et al., Biochem. Biophys. Res. Comm. 163: 1243 (1989) (an allostatin is identified in Diploptera puntata); Chattopadhyay et al. (2004) Critical Reviews in Microbiology 30 (1): 33-54 2004; Zjawiony (2004) J Nat Prod 67 (2): 300-310; Carlini and Grossi-de-Sa (2002) Toxicon, 40 (11): 1515-1539; Ussuf et al. (2001) Cur Sci. 80(7): 847-853; and Vasconcelos and Oliveira (2004) Toxicon 44(4): 385-403. See also U.S. Pat. No. 5,266,317 to Tomalski et al, who disclose genes encoding insect-specific toxins.

(E) An enzyme responsible for a hyperaccumulation of a monoterpene, a sesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivative or another non-protein molecule with insecticidal activity.

(F) An enzyme involved in the modification, including the post-translational modification, of a biologically active molecule; for example, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme, a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, a phosphatase, a ldnase, a phosphorylase, a polymerase, an elastase, a chitinase and a glucanase, whether natural or synthetic. See PCT application WO 93/02197 in the name of Scott et al., which discloses the nucleotide sequence of a callase gene. DNA molecules which contain chitinase-encoding sequences can be obtained, for example, from the ATCC under Accession Nos. 39637 and 67152. See also Kramer et al., Insect Biochem. Molec. Biol. 23: 691 (1993), who teach the nucleotide sequence of a cDNA encoding tobacco hookworm chitinase, and Kawalleck et al., Plant Molec. Biol. 21: 673 (1993), who provide the nucleotide sequence of the parsley ubi4-2 polyubiquitin gene, and U S Pat. Nos. 6,563,020; 7,145,060 and 7,087,810.

(G) A molecule that stimulates signal transduction. For example, see the disclosure by Bolella et al., Plant Molec. Biol. 24: 757 (1994), of nucleotide sequences for mung bean calmodulin cDNA clones, and Griess et al., Plant Physiol. 104: 1467 (1994), who provide the nucleotide sequence of a maize calmodufin cDNA clone.

(II) A hydrophobic moment peptide. See PCT application WO 95/16776 and U.S. Pat. No. 5,580,852 disclosure of peptide derivatives of Tachyplesin which inhibit fungal plant pathogens) and PCT application WO 95/18855 and U.S. Pat. No. 5,607,914 (teaches synthetic antimicrobial peptides that confer disease resistance).

(I) An insect-specific antibody or an immunotoxin derived therefrom. Thus, an antibody targeted to a critical metabolic function in the insect gut would inactivate an affected enzyme, killing the insect. Cf. Taylor et al., Abstract #497, SEVENTH INT'L SYMPOSIUM ON MOLECULAR PLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994) (enzymatic inactivation in transgenic tobacco via production of single-chain antibody fragments).

(I) A virus-specific antibody. See, for example, Tavladoraki et al., Nature 366: 469 (1993), which shows that transgenic plants expressing recombinant antibody genes are protected from virus attack.

(K) A developmental-arrestive protein produced in nature by a pathogen or a parasite. Thus, fungal endo alpha-1,4-D-polygalacturonases facilitate fungal colonization and plant nutrient release by solubilizing plant cell wall homo-alpha-1,4-D-galacturonase. See Lamb et al., Bio/Teclmology 10: 1436 (1992). The cloning and characterization of a gene which encodes a bean endopolygalacturonase-inhibiting protein is described by Toubart et al., Plant J. 2: 367 (1992).

(L) Genes involved in the Systemic Acquired Resistance (SAR) Response and/or the pathogenesis related genes. Briggs, S., Current Biology, 5(2) (1995), Pieterse and Van Loon (2004) Curr. Opin. Plant Bio. 7(4):456-64 and Somssich (2003) Cell 113(7):815-6.

(M) Antifungal genes (Comelissen and Melchers, Pl. Physiol. 101:709-712, (1993) and Parijs et al., Planta 183:258-264, (1991) and Bushnell et al., Can. J. of Plant Path. 20(2):137-149 (1998). Also see U.S. application Ser. Nos. 09/950,933; 11/619,645; 11/657,710; 11/748,994; 11/774,121 and U.S. Pat. Nos. 6,891,085 and 7,306,946.

(N) Detoxification genes, such as for fumonisin, beauvericin, moniliformin and zearalenone and their structurally related derivatives. For example, see U.S. Pat. Nos. 5,716,820; 5,792,931; 5,798,255; 5,846,812; 6,083,736; 6,538,177; 6,388,171 and 6,812,380.

(0) Cystatin and cysteine proteinase inhibitors. See U.S. Pat. No. 7,205,453.

(P) Defensin genes. See W003000863 and U.S. Pat. Nos. 6,911,577; 6,855,865; 6,777,592 and 7,238,781.

(Q) Genes conferring resistance to nematodes. See e.g. PCT Application W096/30517; PCT Application W093/19181, WO 03/033651 and Urwin et al., Planta 204:472-479 (1998), Williamson (1999) Cur Opin Plant Bio. 2(4):327-31; and U.S. Pat. Nos. 6,284,948 and 7,301,069.

(R) Genes that confer resistance to Phytophthora Root Rot, such as the Rps 1, Rps 1-a, Rps lb, Rps 1-c, Rps 14, Rps 1-e, Rps 1-k, Rps 2, Rps 3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps 7 and other Rps genes. See, for example, Shoemaker et al, Phytophthora Root Rot Resistance Gene Mapping in Soybean, Plant Genome 1V Conference, San Diego, Calif. (1995).

(S) Genes that confer resistance to Brown Stem Rot, such as described in U.S. Pat. No. 5,689,035.

(T) Genes that confer resistance to Colletotriehum, such as described in US Patent publication US20090035765. This includes the Reg locus that may be utilized as a single locus conversion.

Seed Treatments and Cleaning Provided are methods of harvesting the grain of the Ful hybrids and using the grain, F2, as seed for sowing. Also provided are methods of using the seed of Ft hybrids, as seed for planting. Embodiments include cleaning the seed, treating the seed, and/or conditioning the seed.

Cleaning the seed includes removing foreign debris such as weed seed, chaff, and non-seed plant matter from the seed. Conditioning the seed can include controlling the temperature and rate of dry down and storing seed in a controlled temperature environment. Seed treatment is the application of a composition to the seed such as a coating or powder. Methods for producing a treated seed include the step of applying a composition to the seed or seed surface. Seeds are provided which have on the surface a composition. Some examples of compositions are insecticides, fungicides, pesticides, antimicrobials, germination inhibitors, germination promoters, cytokinins, and nutrients. Carriers such as polymers can be used to increase binding to the seed.

To protect and to enhance yield production and trait technologies, seed treatment options can provide additional crop plan flexibility and cost-effective control against insects, weeds and diseases. Seed material can be treated, typically surface treated, with a composition comprising combinations of chemical or biological herbicides, herbicide safeners, insecticides, fungicides, germination inhibitors and enhancers, nutrients, plant growth regulators and activators, bactericides, nematicides, avicides and/or raolluscicides. These compounds are typically formulated together with further carriers, surfactants or application-promoting adjuvants customarily employed in the art of formulation. The coatings may be applied by impregnating propagation material with a liquid formulation or by coating with a combined wet or dry formulation. Examples of the various types of compounds that may be used as seed treatments are provided in The Pesticide Manual: A World Compendium, C. D. S. Tomlin Ed., Published by the British Crop Production Council, which is hereby incorporated by reference.

Some seed treatments that may be used on crop seed include, but are not limited to, one or more of abscisic acid, acibenzolar-S-methyl, avermectin, amitrol, azaconazole, azospirillum, azadirachtin, azoxystrobin, Bacillus spp. (including one or more of cereus, firmus, megaterium, pumilis, sphaericus, subtilis and/or thuringiensis), Bradyrhizobium spp. (including one or more of betae, canariense, ellcanii, iriomotense, japonicum, liaonigense, pachyrhizi and/or yuaruningense), captan, carboxin, chitosan, clothianidin, copper, cyazypyr, difenoconazole, etidiazole, fipronil, fludioxonil, fluoxastrobin, fluquinconazole, flurazole, fluxofenim, harpin protein, imazalil, imidacloprid, ipconazole, isoflavenoids, lipochitooligosaccharide, mancozeb, manganese, maneb, mefenoxam, metalaxyl, metconazole, myclobutfinil, PCNB, penflufen, penidilhium, penthiopyrad, permetluine, picoxystrobin, prothioconazole, pyraclostrobin, rynaxypyr, S-metolachlor, saponin, sedaxane, TCMTB, tebuconazole, thiabendazole, thiamethoxam, thiocarb, thiram, toklofos-methyl, triadimenol, trichoderma, trifloxystrobin, triticonazole and/or zinc. PCNB seed coat refers to EPA registration number 00293500419, containing quintozen and terrazole. TCMTB refers to 2-(thiocyanomethyltlaio) benzothiazole.

Seed varieties and seeds with specific transgenic traits may be tested to determine which seed treatment options and application rates may complement such varieties and transgenic traits in order to enhance yield_ For example, a variety with good yield potential but head smut susceptibility may benefit from the use of a seed treatment that provides protection against head smut, a variety with good yield potential but cyst nematode susceptibility may benefit from the use of a seed treatment that provides protection apincr cyst nematode, and so on Likewise, a variety encompassing a transgenic trait conferring insect resistance may benefit from the second mode of action conferred by the seed treatment, a variety encompassing a transgenic trait conferring herbicide resistance may benefit from a seed treatment with a safetter that enhances the plants resistance to that herbicide, etc. Further, the good root establishment and early emergence that results from the proper use of a seed treatment may result in more efficient nitrogen use, a better ability to withstand drought and an overall increase in yield potential of a variety or varieties containing a certain trait when combined with a seed treatment_ In a further embodiment of the invention, products made from seed and plants of the invention include animal fodder in the form of harvested sorghum stems, leaves and grain or parts thereof, sorghum silage, or harvested sorghum stems or parts thereof in dried form, or in a processed dry form such as in the form of pellets, biscuits or cakes.

Embodiments of the invention include different uses of non-seed plant matter such as leaves and stems produced from sorghum plants of the invention or parts thereof as feedstock for a biofuel production process.

A further use for sorghum plant matter of the invention such as leaves and stems is as feedstock for cellulose and/or lignin in the production of paper-based products.

A further use for sorghum plant matter of the invention such as leaves and stems is in the provision of materials for divers industries including the production of fencing; in the manufacture of compacted hardboard; in the production of furniture; in the production of shelving; in the production of insulating material; and in the production of kitchen surfaces.

In a further embodiment of the invention, seeds of sorghum plants of the invention, namely W18, CN7-8 and 1TD6 have been deposited with the NCIMB and provided with deposit numbers NCIMB 43459, NCIMB 43458, and NCIMB 43460, respectively.

There now follow examples and figures further illustrating the invention. It is to be understood that the teaching of the examples and figures is not to be construed as limiting the invention in any way.

FIGURE 1:2011 OP CN7-8 was developed from an initial cross made in The Netherlands of a first selection of NUS28 OP (early) and MN1618 OP (later) populations which displayed successful seed set on the head before harvest. Parent OP lines NUS28 and MN1618 were selected on grain production in the Netherlands and sweetness in the stem.

Two further rounds of selection of NUS28 OP and MN1618 OP populations were conducted before a selective open pollination (SOP) program was conducted using hand crosses made at 12°C, thus inducing male sterility in the seed heads of NUS28, resulting in a 'cold induced male sterility'. The male sterile heads are then crossed with pollen of MN1618 lines.

There followed further SOP steps over six generations using NUS28 SOP and MN1618 SOP populations and crosses of NUS28 lines with MN1618 lines. Then line production was initiated via selfing over at least 3 generations resulting in the line CN7-8. CN7-8 lines have light coloured seeds.

FIGURE 2: 2011 OP W18 was developed from an initial cross made in The Netherlands of a first selection of NUS28 OP (early) and MN1618 OP (later) populations which displayed successful seed set on the head before harvest. Parent OP lines NUS28 and MN1618 were selected on grain production in the Netherlands and sweetness in the stem.

Two further rounds of selection of NUS28 OP and MN1618 OP populations were conducted before a selective open pollination (SOP) program was conducted using hand crosses made at 12°C, thus inducing male sterility in the seed heads of NUS28, resulting in a 'cold induced male sterility'. The male sterile heads are then crossed with pollen of MN1618 lines.

There followed further SOP steps over six generations using NUS28 SOP and MN1618 SOP populations and crosses of NUS28 lines with MN1618 lines. Then line production was initiated via selling over at least 3 generations resulting in the line W18.W18 lines have white seeds.

FIGURE 3:2011 OP III)6 was developed from an initial cross made in The Netherlands of a first selection of NUS28 OP (early) and MN1618 OP (later) populations which displayed successful seed set on the head before harvest. Parent OP lines NUS28 and MN1618 were selected on grain production and sweetness in the stem.

Two further rounds of selection of NUS28 OP and MN1618 OP populations were conducted before a selective open pollination (SOP) program was conducted using hand crosses made at 12°C, thus inducing male sterility in the seed heads of NUS28, resulting in a 'cold induced male sterility'. The male sterile heads are then crossed with pollen of MN1618 lines.

There followed further SOP steps over six generations using NUS28 SOP and MN1618 SOP populations and crosses of NUS28 lines with MN1618 lines. Then line production was initiated via selfing over at least 3 generations resulting in the line H1)6. HD6 lines have red coloured seeds.

Claims (22)

1. CLAIMS1. A method of producing sorghum plants displaying day length neutrality that are propagatable from seed and which display at least one trait selected from i) ulna-earliness; ii) seeds being capable of sprouting and growing into mature plants at latitudes from 30° North up to 64° North (N) in Europe; iii) display cold tolerance in the seed head at latitudes from 30° North up to 55° North (N) in Europe; and iv) being able to mature under low light intensity comprising: a) producing F, sorghum plants in a first generation by hand pollination at diurnal temperatures at or below 12°C wherein each plant displays male sterility and the trait of day length neutrality; b) selecting and crossing open pollinated plants of step a) over at least four generations and selecting plant lines that reliably display at least the selectable trait of day length neutrality in Europe; c) selecting and crossing the plants obtained through step b) with other sorghum plants displaying at least one of the traits selected from i) ultra-earliness; ii) being capable of growing into mature plants at latitudes from 30° North up to 64° North (N) in Europe; iii) display cold tolerance in the seed head at latitudes from 30° North up to 55° North (N) in Europe; and iv) being able to mature under low light intensity; and d) producing sorghum plant populations that, when crossed, will reliably produce a population of plants that substantially uniformly display at least one of the traits i) to iv; e) harvesting the seeds of said crosses; 0 sowing said seeds; and g) growing sorghum plants from the seeds of step f).
2. 2. A method according to claim I, wherein the number of inbreeding steps of step b) is between 4 ant110.
3. 3. A method according to claim 1 or claim 2, wherein sorghum plants capable of growing at latitudes from 30° North up to 64° North are 0 selected having at least a further trait selected from ultra-earliness, cold tolerance in the seed head in plants grown in latitudes from 30° North up to 55° North, high sugar levels in the stem, high levels of antioxidants in the leaves, and high levels of antioxidants in the seed in plants grown in latitudes from 30° North up to 55° North; and ii) inbreeding the sorghum plants until the said further trait or traits of interest is/are stably inherited and wherein the inbred sorghum plants obtained are capable of being reliably reproduced in further generations at latitudes from 30° North up to 55° North.
4. 4. A method according to claim 3, wherein the number of inbreeding steps of step ii) is between 4 and 10.
5. 5. A method according to any one of claims 1 to 4, wherein the parental populations of step c) show substantial uniformity of inheritance for one or more traits selected from the group consisting of day length insensitivity, cold tolerance in the seed head, high sugar levels in the stem, and an ability to set seed at latitudes from 30° North up to 550 North in Europe.
6. 6. The sorghum plants produced according to claim 5.
7. 7. The sorghum plants produced according to claim 5 or 6, wherein said plants display the traits of day length neutrality and cold tolerance in the seed heads at latitudes from 30° North up to 55° North.
8. 8. The sorghum plants produced according to claim 2, wherein the plants are further capable of growing at latitudes from 55° North up to 64° North and display at least the trait of high sugar levels in the stem.
9. 9. Sorghum plants produced according to any one of the preceding claims wherein the sorghum plants are F, hybrids.
10. 10. The seed of the sorghum plants of any one of claims 7 to 9.
11. 11. A method of producing F1 hybrid sorghum plants from the plants obtained from the method of any one of claims 1 to 5 by: a) selecting and crossing a first parental line or clone with a second parental line or a clone that, when crossed, will reliably produce Fi plants that bear seeds; and wherein both parental lines or clones thereof show substantial uniformity in inheritance of characteristics selected from the group consisting of cold tolerant seed heads, high sugar levels in the stem, ability to set viable seed in a wider range of latitudes than that of conventional sorghum; and b) harvesting the seed of said cross; wherein the germination percentage of the seed of b) is at least 60% and gives rise to sorghum plants that produce viable seeds at latitudes from 30° North up to 55° North.
12. 12. The F1 hybrid sorghum seed produced according to the method of claim 11, and plants grown from such hybrid sorghum seed.
13. 13. Sorghum plants that grow at latitudes from 55° North up to 64° North and show substantial uniformity in inheritance of characteristics selected from the group consisting of day length neutrality, high sugar levels in the stem, and increased levels of antioxidants in the leaves.
14. 14. Sorghum plants that grow at latitudes from 30° North up to 55° North and show substantial uniformity in inheritance of characteristics selected from the group consisting of i) day length neutrality ii) high sugar levels in the stem, cold tolerance in the seed head and iv) increased levels of antioxidants in the leaves.
15. 15. Sorghum plants that grow at a latitude from 300 North up to 52° North and show substantial uniformity in inheritance of characteristics selected from the group consisting of i) day length neutrality ii) high sugar levels in the stem, iii) cold tolerance in the seed head, and iv) increased levels of antioxidants in the leaves.
16. 16. Animal fodder produced from sorghum plants or parts thereof as defined in any one of claims 6 to 10, and 12 to 15.
17. 17. Animal fodder according to claim 16 in the form of fresh harvested sorghum stems or parts thereof, sorghum silage, harvested sorghum stems or parts thereof in dried form, or in a processed dry form such as in the form of pellets, biscuits or cakes.
18. 18. Use of non-seed plant matter such as leaves and stems produced from sorghum plants or parts thereof as defined in any one of claims 6 to 10, and 12 to 15 as feedstock for a biofuel production process.
19. 19. Use of sorghum plant matter such as leaves and stems produced from plants or parts thereof as defined in any one of claims 6 to 10, and 12 to 15 as feedstock for cellulose and/or lignin in the production of paper-based products.
20. 20. Use of sorghum plant matter such as dried leaf and stem material produced from plants or parts thereof as defined in any one of claims 6 to 10, and 12 to 15 in the manufacture of compacted hardboard in the production of furniture, shelving, insulating material, and kitchen surfaces.
21. 21. The seed of sorghum plants CN7-8, W18 and 1-11)6 having NCIMB deposit numbers 43458, 43459 and 43460, respectively.
22. 22. Sorghum plants selected from the group CN7-8, W18 and HD6 having NCIMB deposit numbers 43458, 43459 and 43460, respectively.