CN103636505B - A kind of compound selection of high chlorophyll many tillers Fructus Hordei Vulgaris - Google Patents
A kind of compound selection of high chlorophyll many tillers Fructus Hordei Vulgaris Download PDFInfo
- Publication number
- CN103636505B CN103636505B CN201310713460.4A CN201310713460A CN103636505B CN 103636505 B CN103636505 B CN 103636505B CN 201310713460 A CN201310713460 A CN 201310713460A CN 103636505 B CN103636505 B CN 103636505B
- Authority
- CN
- China
- Prior art keywords
- hordei vulgaris
- fructus hordei
- sporidiole
- chlorophyll content
- tiller number
- 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.)
- Active
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention provides the compound selection of high chlorophyll content and/or many stems tiller number Fructus Hordei Vulgaris, prepares the method for barley green product and improve Folium Hordei Vulgaris chlorophyll contents and/or stem tiller number.The method of the present invention includes taking Fructus Hordei Vulgaris microspore development and is in the small ear in monokaryon early stage and/or mid-term, and 2-10 DEG C processes 10-30 days；Results sporidiole, cultivates sporidiole, and regeneration acquisition is isozygotied doubled haploid；With the doubled haploid that isozygotys described in cultivation；Obtain, thus screening, the Fructus Hordei Vulgaris that chlorophyll content is high and/or stem tiller number is many, described Fructus Hordei Vulgaris has been prepared into required barley green product.The present invention by multiple excellent character polymerizations by hybridization technique, is utilized microspore culture that objective trait is quickly isozygotied stable, carries out multi objective screening under human controllable's environment, the final barley resources material obtaining high chlorophyll content many stems tiller.The elite germplasm material that this material can be produced as nutraceutical product such as barley greens by field planting or water planting mode.
Description
Technical field
The invention belongs to agricultural technology field, be specifically related to a kind of high chlorophyll content and the screening breeding method of many stems tiller barley material.
Background technology
The seedling of wheat crops is a kind of Chinese traditional herbs, China's medical value describing wheat seedling in Compendium of Material Medica and " typhoid fever summary ".1969, Japanese scholars Miscanthus sacchariflorus (Maxim) Benth et Hook f original meaning show doctor delivered " grass greenery composition and as stable medicine, food research " literary composition, and is devoted to commercialized development.1991, he utilized the technology of " ultralow temperature fast spraying seasoning ", is squeezed the juice by Herba Hordei Vulgaris and is condensed into powder, is called " barley green " (BarelyGreen), and promoted to international market.Barley green product in the market is for primary raw material, to be aided with what the dispensings such as dextrin processed with the tender leaf (wheat height 20-30cm) of the wheat crops tillering regularity such as Fructus Hordei Vulgaris or Semen Tritici aestivi.Barley green rich in proteins, chlorophyll, substantial amounts of organized enzyme (such as SOD enzyme, catalase, amylase, protease etc.), beta-carotene, multivitamin, and various trace elements (Guo Jianhua, 2008).
Modern biotechnology test and clinical research confirmation, barley green has following 9 aspect physiological functions: (1) promotes the restitution of damage dna；(2) anti-inflammatory analgesic；(3) enhancing immunity and anti-cancer, anticancer, radiation resistance；(4) HIV (human immunodeficiency virus) effect is suppressed；(5) blood pressure lowering；(6) antiulcer；(7) unnecessary free radical in purged body, defying age and preventing and treating cardiovascular and cerebrovascular disease；(8) diabetes are prevented and treated；(9) accelerating wound healing, promotes hemopoietic, eliminates halitosis (Qiao Wenjing, 2009).U.S. FDA approved yong barley leaves juice is as food supplement.In Japan, yong barley leaves juice product has obtained the mark (Niu Guangcai, 2011) of the health food that health association of Japan is assert.Abroad wheat leaf series of products are developed at present, such as Semen Tritici aestivi fiber food, Folium Hordei Vulgaris short health food, wheat straw leaf beverage, barley green wheat enzyme nutriment laughable, blue or green etc., Japan is also proposed in yong barley leaves juice powder to add the tonic food of dextrin, yeast, Radix Dauci Sativae powder, Korean Ginseng powder.
Barley green product has good alimentary health-care function, there is wide market prospect, the major production base of barley green is in the U.S., Canada, Australia etc., pure natural property and health-care effect due to it, cater to the pursuit of people's " back to nature ", thus fashionable North America, Europe, Australia, Southeast Asia, every annual sales amount has reached 23～2,700,000,000 dollars (yellow prime ministers, 2003).The production of Fructus Hordei Vulgaris functional product will increase the added value of barley cultivation industry, makes increasing peasant income, it was reported that, every mu can produce fresh leaf 500kg, the output value is up to more than 1000 yuan, and farmers' income exceeds more than one times (Wu Hongxia, 2002) than the conventional Fructus Hordei Vulgaris of plantation and other winter crops.
But the special barley variety producing barley green at present lacks, and seriously limits the development and utilization of barley green product.The selection-breeding of current function Fructus Hordei Vulgaris does not also cause enough attention in China, and traditional breeding objective can not meet the requirement of function Fructus Hordei Vulgaris selection-breeding.Traditional breeding objective mainly considers economical character such as Grain yield and quality, the disease resistance etc. of kind, and takes from vegetative growth phase producing the special barley variety raw material of barley green, accordingly, it would be desirable to rethink its breeding goal.To Barley Varieties Applied for Barley Green selection-breeding, it is desirable to the nutrient contents such as seedling growth is fast, and during jointing, vine yield is high, chlorophyll are high.
Blade is that plant carries out photosynthetic vitals, and chlorophyll is plant carries out photosynthetic important pigment, its effect is just by luminous energy and catches, the height of its content not only can affect leaf color, and photosynthetic efficiency can be directly affected, there are some researches show that chlorophyllous content is to become positively related with photosynthetic efficiency within the specific limits.Green plants is all dependent on photosynthesis and carries out biosynthesis, completes its morphogenesis and biomass accumulation.The chlcrophyll biosynthesis of green plants is a complex process being participated in synthesis by a series of enzymes, and the enzyme and the gene that wherein participate in synthesis are clearer in higher plant, but the hereditary pattern controlling chlorophyll content is not clear.Screening to chlorophyll content at present is all concentrate on land for growing field crops screening, adopts chlorophyll meter or chlorophyll fluorescence instrument to measure on the spot.There are some problems in this screening mode, there are some researches show that chlorophyllous content is effected by environmental factors substantially, the content of different growth periods also can change, land for growing field crops circumstance complication, planting conditions is subject to various factors impact and restriction, and chlorophyll content performance is also inconsistent, and additionally chlorophyll is largely subject to recessive gene control, screening needs to carry out in high generation, and this obtains, by needing the longer time, offspring of isozygotying.
Except screening high chlorophyll content, its overground part biological yield is also an important indicator.Tiller stem and leaf is the biological yield composition part of aerial parts in seedling stage, will improve its biological yield, it is necessary to selection tiller is many, the eugonic material of stem and leaf.The screening of Barley Breeding has focused largely on the growth and maturity phase, and the overground part biological yield in seedling stage and the screening of chlorophyll content be have not been reported.
The present invention will provide for a kind of some problems overcoming current land for growing field crops to screen, enforcement is effective, the method for the Fructus Hordei Vulgaris of quick breeding high chlorophyll content many stems tiller, including: by hybridization technique by multiple excellent character polymerizations, utilize microspore culture that objective trait is quickly isozygotied stable, multi objective screening is carried out, the final barley resources material obtaining high chlorophyll content many stems tiller under human controllable's environment.The elite germplasm material that this material can be produced as nutraceutical product such as barley greens by field planting or water planting mode.
Summary of the invention
It is an object of the invention to the Fructus Hordei Vulgaris efficient, high chlorophyll content, many stems tiller number is quickly provided, and the described character of described Fructus Hordei Vulgaris can stable heredity.The present invention is realized by the following method this purpose, and described method has stability and repeatability.
One aspect of the present invention provides the method for the Fructus Hordei Vulgaris of high chlorophyll content and/or many stems tiller number to include:
(1) take Fructus Hordei Vulgaris microspore development and be in the small ear in monokaryon early stage and/or mid-term, K cryogenic treatment 2-3 week；
(2) results sporidiole, cultivates sporidiole, and regeneration acquisition is isozygotied doubled haploid；
(3) isozygoty described in water planting doubled haploid；
The Fructus Hordei Vulgaris that chlorophyll content is high and/or stem tiller number is many is obtained with screening.
Another aspect of the present invention provides a kind of method preparing barley green product, and described method includes:
(1) take Fructus Hordei Vulgaris microspore development and be in the small ear in monokaryon early stage and/or mid-term, K cryogenic treatment 2-3 week；
(2) results sporidiole, cultivates sporidiole, and regeneration acquisition is isozygotied doubled haploid；With
(3) isozygoty described in water planting doubled haploid, and screening obtains the Fructus Hordei Vulgaris that chlorophyll content is high and/or stem tiller number is many；With
(4) described Fructus Hordei Vulgaris is prepared into barley green product.
Further aspect of the present invention provides a kind of method improving Folium Hordei Vulgaris chlorophyll contents and/or stem tiller number, and described method includes:
(1) taking Fructus Hordei Vulgaris microspore development and be in the small ear in monokaryon early stage and/or mid-term, 2-10 DEG C processes 10-30 days；
(2) results sporidiole, cultivates sporidiole, and regeneration acquisition is isozygotied doubled haploid；
(3) isozygoty described in water planting doubled haploid, it is thus achieved that regeneration plant；
Wherein, described regeneration plant is the plant that chlorophyll content improves and/or stem tiller number increases.
Should be understood that described " chlorophyll content raising " and " stem tiller number increases " are for initial barley material, for the chlorophyll content of the Fructus Hordei Vulgaris namely obtained without the inventive method process relative to the described Fructus Hordei Vulgaris sporidiole of step (1) and stem tiller number.
In a specific embodiment, said method of the present invention also included before step (1):
A () chooses the barley material that a collection of leaf color is dark green, seedling growth is vigorous, tiller is more, the chlorophyll content of 2 leaf 1 heart stages is measured after water planting, start to add up tiller number, plant height and/or basal part of stem girth to tillering stage, filter out chlorophyll content height according to these statistical indicators, tiller number is many, grow rapid kind；With
B () cultivates described kind, it is thus achieved that the sporidiole described in step (1)；With
If c () barley material only has chlorophyll content height, tiller number is many, the objective trait grown in rapidly, then mutually pollinate after pollen maturation hybridization, obtain hybridization F0 for seed, breed this F0 for seed, obtain F1 generation plant, obtain the sporidiole described in step (1) from this F1 generation plant.
In a specific embodiment, in said method of the present invention, sporidiole is available from the Fructus Hordei Vulgaris that chlorophyll content is high and stem tiller number is many.
In a specific embodiment, in said method of the present invention, described step (2) including:
(2a) sterilize described tassel, then clean；
(2b) sporidiole is separated from described tassel；
(2c) in room temperature, dark processing sporidiole 36-60 hour；
(2d) suspend sporidiole with inducing culture, and by inoculation of suspension liquid in inducing culture, callus induction；With
(2e) induction gained callus proceeding to division culture medium, cultivate and obtain regrowth, isozygoty described in namely doubled haploid.
In a specific embodiment, in said method of the present invention, when separating sporidiole, test tube adds tassel and the CaCl containing 4-8% mannitol, 1.0-1.5g/L2With the extracting solution of 2-(N-morpholine) ethyl sulfonic acid of 0.8-1.2g/L, exceed the speed limit rotary-cut with high speed disperser, filter, centrifugal filtrate, thus obtaining sporidiole.
In a specific embodiment, in said method of the present invention, inducing culture is with N6 culture medium for minimal medium, wherein added with 6-Furfurylaminopurine 0.3-0.8mg/L, 2,4-dichlorphenoxyacetic acid 0.5-1.5mg/L and maltose 70-120g/L；Division culture medium is with MS for minimal medium, wherein added with 6-benzyl aminoadenine 0.3-0.8mg/L, KT1.0-2.0mg/L, naphthalene acetic acid 0.03-0.08mg/L and maltose 15-50g/L, solidifies with 3-8g/L agar powder.
In a specific embodiment, in said method of the present invention, also include after step (3):
The regeneration plant that microspores culture obtains is put in Fructus Hordei Vulgaris standard liquid nutrient and cultivates, again screen, select the Fructus Hordei Vulgaris that chlorophyll content is high and/or stem tiller number is many, and collect its seed.
In a specific embodiment, described screening again is to adopt water planting mode to screen under human controllable's environment.
In a specific embodiment, described human controllable's environment includes: adopting fluorescent tube illumination in the controlled environment chamber, intensity of illumination is about 1500-2500lx, it is preferable that about 1800-2000lx；Keep room temperature be about 18-22 DEG C daytime, it is preferable that about 20 DEG C, be about 16-20 DEG C in the evening, it is preferable that 18 DEG C, daytime, night temperature fluctuation be about 1-2 DEG C.
In a specific embodiment, described barley green product is selected from: barley fiber food, Folium Hordei Vulgaris urge health food, wheat straw leaf beverage, barley green wheat enzyme nutriment laughable, blue or green, and add the tonic food of dextrin, yeast, Radix Dauci Sativae powder, Korean Ginseng powder in yong barley leaves juice powder.
In a specific embodiment, in said method of the present invention, the sporidiole obtained is with pretreatment fluid in room temperature, dark pretreatment 36-60 hour, and wherein, described pretreatment fluid contains the CaCl of 4-8% mannitol, 1.0-1.5g/L22-(N-morpholine) ethyl sulfonic acid with 0.8-1.2g/L.
Preferably, the present invention obtains barley strain chlorophyll content and stem tiller number all improve.
The present invention provides method to have the feature that
1. barley green products material is drawn materials and is usually in the 6-7 leaf phase, and therefore the water planting in 6-8 week can complete incubation, reaches predetermined growth period, and period can conveniently be measured and draw materials.The kind that character is excellent is filtered out respectively according to statistical indicator described herein.What the present invention adopted is the unified condition of culture of human controllable, completes in the controlled environment chamber, and not by controlling season, screening has stability and repeatability.2 leaf 1 heart stages were highstrung screening periods, and interracial comparison in difference is obvious, and unconspicuous kind can also compare in the METHOD FOR CONTINUOUS DETERMINATION in later stage.Select tillering number as an index evaluating the upperground part biomass simultaneously.
2. screening target carries out around Parameters of Barley Seedling chlorophyll content and two targets of aerial parts biological yield, does not investigate after material the character such as period of duration grain yield.
3. pair Preliminary screening containing better objective trait barley material, adopt microspore culture to obtain doubled haploid (DH) strain of isozygotying, plant all character of matter and isozygoty completely, recessive character can show the present age.
4. the barley material of pair single excellent objective trait adopts hybridization technique aggregate target character.
5. selecting different index to screen in a large number according to objective trait, screening relates to first material selection, DH plant of isozygotying, hybridization polymerization trait segregation offspring.
6. the usual 2-3 consuming time of existing land for growing field crops triage techniques, even longer time, it is possible to screen the barley material that can be used to prepare barley green product obtaining that there is above-mentioned objective trait, but screen deficient in stability and repeatability.Due to the fact that and the special handling of sporidiole is substantially shorter this screening time, generally can realize in 1 year.
Detailed description of the invention
The above-mentioned method of the present invention includes:
(1) take Fructus Hordei Vulgaris microspore development and be in the small ear in monokaryon early stage and/or mid-term, K cryogenic treatment 10-30 days；
(2) results sporidiole, cultivates sporidiole, and regeneration acquisition is isozygotied doubled haploid；With
(3) isozygoty described in water planting doubled haploid.
Described " low temperature " is often referred to 2-10 DEG C, for instance 3-5 DEG C.K cryogenic treatment refers to place at low temperatures the described small ear of fresh harvesting.The time of K cryogenic treatment is usually 10-30 days, for instance can be 14-21 days not etc..
Sporidiole is normally taken from chlorophyll content height, tiller number is many, grow rapid parent material seed plants the Fructus Hordei Vulgaris obtained.
Herein, " chlorophyll content high " refer to regard to screen batch for, the plant that chlorophyll content is of a relatively high, for instance, refer to the strain of 20% before being screened batch Determination of Chlorophyll content and coming, it is preferable that front 10%, more preferably front 5%.Certainly several strains that chlorophyll content is the highest or relatively the highest it are selected from.
Herein, " tiller number is many " refer to regard to screen batch for, the how relatively large number of plant of tiller number, for instance, refer in being screened batch tiller number come before 20% strain, it is preferable that front 10%, more preferably front 5%.
Herein, " growth rapidly " with regard to screen batch for, the comparatively faster plant of the speed of growth, for instance, refer in the being screened batch speed of growth come before 20% strain, it is preferable that front 10%, more preferably front 5%.
Generally, can first according to objective trait, choose the barley material that a collection of leaf color is dark green, seedling growth is vigorous, tiller is more, the chlorophyll content of 2 leaf 1 heart stages is measured after water planting, start to add up tiller number, plant height and/or basal part of stem girth to tillering stage, the kind that character is excellent is filtered out respectively, for instance filter out chlorophyll content height, tiller number is many, grow rapid kind according to these statistical indicators.
The standard Fructus Hordei Vulgaris mill water culture nutrient solution (improvement Hoagland solution) reported can be adopted to carry out water planting, and this nutrient source formula is as follows:
NH4NO3(0.2mM)、KNO3(5mM)、CaNO3.2(2mM)、MgSO4(2mM)、KH2PO4(0.1mM)、Na2SiO3(0.5mM)、NaFe(III)–EDTA(0.05mM)、H3BO3(0.01mM)、MnCl2(0.005mM)、ZnSO4(0.005mM)、CuSO4(0.0005mM)、Na2MoO3(0.0001mM) front solution ph dilute hydrochloric acid, is used to regulate to about 6.0.
Water planting can in the controlled environment chamber in carry out.Employing fluorescent tube illuminates, and intensity of illumination is about 2000lx, keeps room temperature daytime 20 degree, evening 18 degree, temperature fluctuation 1-2 degree.
Then, plant the seed (i.e. parent material seed) of this kind, draw materials for microspores culture.Can plant big Tanaka, or, if plant screening material is few, it is possible to plant in the controlled environment chamber after screening.
For the material that only one of which objective trait is excellent, hybridization (positive and negative hybridization) of can mutually pollinating after pollen maturation, it is thus achieved that F0 is for seed in hybridization.Land for growing field crops or phjytotron plantation F0 are for seed, it is thus achieved that F1 generation plant, draw materials for microspores culture.
The chlorophyll content can planted from land for growing field crops or phjytotron is high, tiller number is many, grow little Hua microspore development in the middle part of choosing in rapid kind is in monokaryon early stage, the small ear in mid-term carries out K cryogenic treatment 10-30 days, as mentioned before.
After K cryogenic treatment, technical limit spacing sporidiole known in the art can be adopted.Including sterilization, high speed centrifugation etc..
Herein, sterilizing typically by 75% alcohol-pickled 20-60 second, saturated Eusol is sterilized 10-20 minute.Typically by 75% alcohol-pickled 30 seconds, saturated Eusol was sterilized 15 minutes.Those skilled in the art's other sterilization means known.
After sterilization, available aseptic water washing is for several times, for instance 3-4 time.Then, each test tube connects 5-15 tassel, for instance often 10 tassels of pipe.Pour 10-20ml, preferred 15ml extracting solution into, exceed the speed limit rotary-cut with high speed disperser, filter (such as, filter with 150 eye mesh screens), centrifugal filtrate (such as, with the centrifugal 5min of 100 × g, repeat 3 times), collection sporidiole.Extracting solution preferably comprises the CaCl of 4-8% mannitol, 1.0-1.5g/L2MES(2-(N-morpholine) ethyl sulfonic acid with 0.8-1.2g/L).In a specific embodiment, the CaCl of extracting solution preferably 6% mannitol, 1.1g/L2MES with 0.976g/L.
Optionally with pretreatment fluid in room temperature (such as 25 DEG C), dark pretreatment 36-60 hour, generally process 2 days.The same extracting solution of composition of pretreatment fluid.
Then, optionally by 21%(mass percent by volume) maltose purifies sporidiole, then washs 1 time with inducing culture, then with inducing culture by sporidiole Auto-regulating System of Density of Heavy Medium to 0.5-2.0 × 105/ ml, it is preferable that 1.0 × 105/ ml, takes about 1ml microspore suspension and is inoculated in culture dish (such as, in the culture dish of 35 × 15mm), room temperature (25 DEG C) light culture, callus induction after sealing；Then proceed to division culture medium and obtain regrowth.Obtain plant is the doubled haploid that isozygotys (DH) that single cell culture obtains herein.The monoploid detected can directly double with colchicine treatment to become doubled haploid.
Inducing culture is with N6Culture medium is minimal medium, wherein added with 6-Furfurylaminopurine (KT) 0.3-0.8mg/L, 2,4-dichlorphenoxyacetic acids (2,4-D) 0.5-1.5mg/L and maltose 70-120g/L.In a detailed description of the invention, inducing culture is added with KT0.4-0.6mg/L and 2,4-D0.8-1.2mg/L, and maltose 80-100g/L.In a detailed description of the invention, inducing culture is added with KT0.5mg/L and 2,4-D1.0mg/L, and maltose 90g/L.
Conventional N6The composition of culture medium is substantially as shown in table 1 below, and the concentration of different vendor or the heterogeneity described in document perhaps has little difference, but its function is all roughly the same:
Table 1:N6The formula (unit: mg/L) of culture medium:
| N6A great number of elements: | |
| (NH4)2SO4 | 463 5 --> |
| KNO3 | 2830 |
| CaCl2·2H2O | 166 |
| MgSO4·7H2O | 185 |
| KH2PO4 | 400 |
| N6Trace element: | |
| H3BO3 | 1.6 |
| KI | 0.83 |
| MnSO4·4H2O | 4.4 |
| ZnSO4·7H2O | 1.5 |
| N6Iron salt: | |
| Na2EDTA | 37.3 |
| FeSO4·7H2O | 27.8 |
| N6Organic element: | |
| Thiamine hydrochloride | 1.0 |
| Pyridoxine Hydrochloride | 0.5 |
| Glycine | 2.0 |
| Nicotinic acid | 0.5 |
Voluntarily by the inducing culture of the above-mentioned formula preparation present invention, or can buy acquisition N to chemical reagents corporations such as Sigma6Culture medium finished product.Then the inducing culture according to formula described herein preparation this paper.It will be appreciated by those skilled in the art that, concentration shown in above-mentioned table 2 can be made suitable variation, such as have about 5% or lower (such as about 3%, about 2%, about 1%, or less mobility scale) variation, thus prepare the culture medium obtained and still there is required biological function, remain to for implementing the present invention.Such as, with KH2PO4For example, every liter of N6Culture medium can contain the KH of about 380-420mg2PO4(amplitude of fluctuation of 5%).The concentration of other composition also so changes.Additionally, N6Composition in culture medium is used as function and the same or analogous composition of character is replaced.
Division culture medium is with MS for minimal medium, wherein added with 6-benzyl aminoadenine (6-BA) 0.3-0.8mg/L, KT1.0-2.0mg/L, naphthalene acetic acid (NAA) 0.03-0.08mg/L and maltose 15-50g/L, solidifies with 3-8g/L agar powder.
In a detailed description of the invention, division culture medium is added with 6-benzyl aminoadenine (6-BA) 0.4-0.6mg/L, KT1.3-1.8mg/L, naphthalene acetic acid (NAA) 0.03-0.06mg/L and maltose 20-40g/L, solidifies with 4-6g/L agar powder.
In a detailed description of the invention, division culture medium is added with 6-BA0.5mg/L, KT1.5mg/L, NAA0.05mg/L and maltose 30g/L, solidifies with 6g/L agar powder.
The formula of MS culture medium is as shown in table 2 below (unit: mg/L):
Table 2
| MS a great number of elements: | |
| NH4NO4 | 1650 |
| KNO3 | 1900 |
| CaCl2·2H2O | 440 |
| MgSO4·7H2O | 370 |
| KH2PO4 | 170 |
| MS trace element: | |
| H3BO3 | 6.2 |
| KI | 0.83 |
| MnSO4·4H2O | 22.3 |
| ZnSO4·7H2O | 8.6 |
| Na2MoO4·2H2O | 0.25 |
| CuSO4·5H2O | 0.025 |
| CoCl2·6H2O | 0.025 |
| MS iron salt: | |
| Na2EDTA | 37.3 |
| FeSO4·7H2O | 27.8 |
| MS organic element: | |
| Thiamine hydrochloride | 0.1 |
| Pyridoxine Hydrochloride | 0.5 |
| Nicotinic acid | 0.5 |
| Glycine | 2.0 |
| Inositol | 100.0 |
MS can voluntarily by the preparation of above-mentioned formula, it is possible to buy finished product to chemical reagents corporations such as Sigma.It will be appreciated by those skilled in the art that, concentration shown in above-mentioned table 2 can be made suitable variation, such as have about 5% or lower (such as about 3%, about 2%, about 1%, or less mobility scale) variation, thus prepare the culture medium obtained and still there is required biological function, remain to for implementing the present invention.Such as, for inositol, every liter of MS culture medium can contain the inositol (amplitude of fluctuation of 5%) of about 95-105mg.The concentration of other composition also so changes.Composition in inducing culture is used as function and the same or analogous composition of character is replaced.
Being removed from culture medium by the regeneration plant (when Length of shoot and root is more than 2cm) that microspores culture obtains, clear water cleans root agar, wraps up foundation portion with sponge strip, it is placed on floating plate, putting in Fructus Hordei Vulgaris standard liquid nutrient and cultivate, phjytotron controls growing environment condition, again screens.The material with excellent character that this time screening obtains can stablize heredity, and can screen the target material that in single part of material, two character are all more satisfactory.The plant screened can transplant the cultivation of basin alms bowl earth, and phjytotron plantation, to Grain Ripening, is gathered in the crops single-strain seed, ibid bred conservation.
Optionally, described seed can be planted, thus gathering in the crops the material of high chlorophyll content, many stems tiller.
The condition of water planting, including illumination, temperature, culture fluid etc. as mentioned before.
By cultivating the regeneration plant that sporidiole obtains, this material is the doubled haploid (DH) isozygotied by a set of chromosome doubling, and the hereditary character of parent is isozygotied completely, and recessive character can in plant performance in the present age.
Therefore, the present invention also includes the barley strain and the seed thereof that adopt the inventive method to prepare, and adopt the barley green product that this barley strain prepares, include but not limited to that barley fiber food, Folium Hordei Vulgaris urge health food, wheat straw leaf beverage, barley green wheat enzyme nutriment laughable, blue or green, and in yong barley leaves juice powder, add the tonic food etc. of dextrin, yeast, Radix Dauci Sativae powder, Korean Ginseng powder.
Hereafter by the way of specific embodiment, present invention is described.Should be understood that following example are only illustrative, the present invention can be made various modifications and changes when not necessarily departing from spirit and scope of the invention.The scope of the present invention is limited by claims hereof.Furthermore, it is to be understood that in above-mentioned each culture medium the preferable range of each composition can combination in any, as long as it can realize the goal of the invention of the present invention.
Embodiment
Embodiment 1: parental line selection
Estimate situation according to field planting barley material, choose the material 200 parts that leaf color is dark green, seedling growth is vigorous, tiller number is many, point individual plant results seed.Take 100, seed, surface adopts 75% ethanol to soak 30 seconds, 1% hypochlorite disinfectant 15 minutes, distilled water flushing is clean, is placed on moistening gauze in the accelerating germination of room temperature dark place overnight, robust growth is chosen after germination, seedling 30 strain more than 1 centimetre of the radical bud length, wraps up seed with sponge strip, and (floating plate usable foam plate punches by 5 centimetres of spacing to be placed in floating plate, diameter about 1.5 centimetres) in circular hole, it is placed in nutritional solution and cultivates.Nutritional solution adopt reported standard Fructus Hordei Vulgaris mill water culture nutrient solution (improvement Hoagland solution, formula is as follows:
NH4NO3(0.2mM);KNO3(5mM);CaNO3.2(2mM);MgSO4(2mM);KH2PO4(0.1mM);Na2SiO3(0.5mM);NaFe(III)–EDTA(0.05mM);H3BO3(0.01mM);MnCl2(0.005mM);ZnSO4(0.005mM);CuSO4(0.0005mM);Na2MoO3(0.0001mM). before using, solution ph dilute hydrochloric acid regulates to about 6.0.
Phjytotron adopts fluorescent tube illumination, and intensity of illumination is about 2000lx, and large-size air conditioning temperature control is blown, and keeps room temperature daytime 20 degree, evening 18 degree, temperature fluctuation 1-2 degree.
When Seedling grew to for 21 heart stage of leaf, measuring the 2nd leaf, each plant METHOD FOR CONTINUOUS DETERMINATION central region 10 times with Japan's SPAD-502 hand-held chlorophyll meter, read meansigma methods as chlorophyll content value, this, storeroom comparison in difference was obvious in period.Later each week measures once 2 leaves such as method, terminates (when growing to 6-7 leaf) to whole cultivation.After tiller starts, add up weekly tiller number, measure plant height simultaneously and lobe numbers evaluates its speed of growth.
After cultivation terminates, all material determination data is carried out statistical analysis, find out the material with significant difference.
Embodiment 2: screening parent's plantation
Selection has the material of above-mentioned excellent character one or several (growth is rapidly for high chlorophyll, many stems tiller) as the parent material for hybridizing, field seeding then.Select size uniformly, the planting seed of full seed in land for growing field crops, spacing 3cm, line-spacing 30cm.Containing organic 34.0g/kg, full nitrogen 2.41g/kg, available nitrogen 37.30mg/kg in soil, basal dressing 20kg/ mu before sowing, tillering stage 10kg/ mu carbamide, period can its leaf normal complexion overground part biological yield of sampling Detection.
Embodiment 3: hybridization
The material carrying different target character (high chlorophyll or many tillers) by two parts carries out positive and negative hybridization, concrete grammar is when pollen maturation, cuts off maternal plant tassel grain husk shell, takes paternal plant little Hua pollen and pollinates, carry out listing mark simultaneously, after maturation, individually gather in the crops hybrid seed.
Embodiment 4:F1 is for plant breeding
Hybrid seed ibid in sowing season kind in land for growing field crops, it is thus achieved that F1 generation growth plant, as microspore-isolated culture material.
Embodiment 5: microspores culture obtains DH plant of isozygotying
Choose from big Tanaka that middle part little Hua microspore development is in monokaryon early stage, the small ear in mid-term carries out 5 degree of K cryogenic treatment 2-3 week, during inoculation, tassel was with 75% alcohol-pickled 30 seconds, saturated Eusol is sterilized 15 minutes, aseptic water washing 3-4 time, each test tube connects 10 tassels, pour 15ml extracting solution into (containing 6% mannitol, 1.1g/LCaCl2And 0.976g/LMES), to exceed the speed limit rotary-cut with high speed disperser, filter with 150 eye mesh screens, filtrate, with the centrifugal 5min of 100 × g, is repeated 3 times, collection sporidiole.
Sporidiole with pretreatment fluid in 25 DEG C, dark pretreatment 2d.
Before cultivating, sporidiole is first purified with 21% maltose, then wash 1 time with inducing culture, then with inducing culture by sporidiole Auto-regulating System of Density of Heavy Medium to 1.0 × 105/ ml, takes 1ml microspore suspension and is inoculated in culture dish (35 × 15mm), and Parafilm film seals, 25 DEG C of light culture, callus induction.
Inducing culture is with the N of improvement6Culture medium is minimal medium, wherein added with KT0.5mg/L and 2, and 4-D1.0mg/L, maltose 90g/L.
Callus is proceeded to division culture medium and obtains regrowth.Division culture medium is with MS for minimal medium, wherein added with 6-BA0.5mg/L, KT1.5mg/L, NAA0.05mg/L and maltose 30g/L, solidifies with 6g/L agar powder.
By cultivating the plant obtaining sporidiole regeneration, this material is the doubled haploid (DH) isozygotied by a set of chromosome doubling, and the hereditary character of parent is isozygotied completely, and recessive character can in plant performance in the present age.
Can ibid carry out water planting and again screen the barley material obtaining two objective traits polymerization (high chlorophyll content and many tillers).
Embodiment 6:DH plant is screened
The regeneration plant (when Length of shoot and root is more than 2cm) that microspores culture obtains is removed from culture medium, clear water cleans root agar, foundation portion is wrapped up with sponge strip, it is placed on floating plate, put in Fructus Hordei Vulgaris standard liquid nutrient and cultivate, phjytotron controls growing environment condition, and the same step is screened again.The material with excellent character that this time screening obtains can stablize heredity, and can screen the target material (two characteristic indexs are shown in following table) that in single part of material, two character are all more satisfactory.The plantlet of transplant basin alms bowl earth cultivation screened, phjytotron plantation, to Grain Ripening, is gathered in the crops single-strain seed, is ibid bred conservation.
Choose the seed being unscreened roguing system (strain) as comparison of the original strain without hybridization and microspores culture, germinate by preceding method, carry out after seedling adopting under identical mill water culture nutrient solution and artificial climate growth conditions, be measured in the same method chlorophyll and individual plant stem tiller number index compares.
Table 3 below shows chlorophyll relative content (adopting the test of SPAD method) and the individual plant stem tiller number of the different strains adopting the inventive method and contrast method to obtain.
Table 3
The result of table 3 shows, exceeds all far away the comparison barley material processed without the inventive method through the barley material chlorophyll content of the inventive method selection-breeding and stem tiller number.Such as, for chlorophyll content, the chlorophyll content of Fructus Hordei Vulgaris of the present invention has at least the raising (comparing strain L3 and strain L10) of about 20% relative to the chlorophyll content of comparison Fructus Hordei Vulgaris, and the highest about 85%(that improves compares strain L2 and L8)；Equally, the stem tiller number of Fructus Hordei Vulgaris of the present invention has at least the increase (compare strain L3/L4 and strain L6, and the highest about 230%(that increases compares strain L1 and strain L7) of about 17% relative to the stem tiller number of comparison Fructus Hordei Vulgaris.And, from the data of table 3 it can be seen that the chlorophyll content of Fructus Hordei Vulgaris processed through the inventive method and stem tiller number generally all be higher than the chlorophyll content compareing Fructus Hordei Vulgaris and stem tiller number.
Claims (13)
1. the method producing the Fructus Hordei Vulgaris that chlorophyll content improves and/or stem tiller number increases, described method includes:
(1) taking Fructus Hordei Vulgaris microspore development and be in the small ear in monokaryon early stage and/or mid-term, 2-10 DEG C processes 10-30 days；
(2) results sporidiole, cultivates sporidiole, and regeneration acquisition is isozygotied doubled haploid；
(3) isozygoty described in water planting doubled haploid；With
(4) screening obtain the chlorophyll content of Fructus Hordei Vulgaris and stem tiller number that process without abovementioned steps (1)-(3) and obtain relative to the described Fructus Hordei Vulgaris sporidiole of step (1) for chlorophyll content improves and/or stem tiller number increases Fructus Hordei Vulgaris；
Wherein, described step (2) including:
(2a) sterilize described tassel, then clean；
(2b) sporidiole is separated from described tassel；
(2c) processing sporidiole 36-60 hour in room temperature, dark pretreatment fluid, wherein, described pretreatment fluid contains the CaCl of 4-8% mannitol, 1.0-1.5g/L22-(N-morpholine) ethyl sulfonic acid with 0.8-1.2g/L；
(2d) suspend sporidiole with inducing culture, and by inoculation of suspension liquid in inducing culture, callus induction；With
(2e) induction gained callus proceeding to division culture medium, cultivate and obtain regrowth, isozygoty described in namely doubled haploid；
Wherein, in step (2b), when separating sporidiole, test tube adds tassel and extracting solution, exceed the speed limit rotary-cut with high speed disperser, filter, centrifugal filtrate, thus obtaining sporidiole, wherein, described extracting solution contains the CaCl of 4-8% mannitol, 1.0-1.5g/L22-(N-morpholine) ethyl sulfonic acid with 0.8-1.2g/L；
Wherein, described inducing culture is with N6Culture medium is minimal medium, wherein added with 6-Furfurylaminopurine 0.3-0.8mg/L, 2,4-dichlorphenoxyacetic acid 0.5-1.5mg/L and maltose 70-120g/L；
Described division culture medium is with MS for minimal medium, wherein added with 6-benzyl aminoadenine 0.3-0.8mg/L, KT1.0-2.0mg/L, naphthalene acetic acid 0.03-0.08mg/L and maltose 15-50g/L, solidifies with 3-8g/L agar powder.
2. the method for claim 1, it is characterised in that described method also included before step (1):
A () chooses the barley material that a collection of leaf color is dark green, seedling growth is vigorous, tiller is more, the chlorophyll content of 2 leaf 1 heart stages is measured after water planting, start to add up tiller number, plant height and/or basal part of stem girth to tillering stage, filter out chlorophyll content height according to these statistical indicators, tiller number is many, grow rapid kind；With
B () cultivates described kind, it is thus achieved that the sporidiole described in step (1)；With
If c () barley material only has chlorophyll content height, tiller number is many, the objective trait grown in rapidly, then mutually pollinate after pollen maturation hybridization, obtain hybridization F0 for seed, breed this F0 for seed, obtain F1 generation plant, obtain the sporidiole described in step (1) from this F1 generation plant；
Wherein, the kind that described chlorophyll content is high refers to the strain of 20% before being screened batch Determination of Chlorophyll content and coming；The kind that described tiller number is many refer in being screened batch tiller number come before 20% strain；The rapid kind of described growth refer in the being screened batch speed of growth come before 20% strain.
3. the method for claim 1, it is characterised in that
Inducing culture is with N6Culture medium is minimal medium, wherein added with 6-Furfurylaminopurine 0.4-0.6mg/L, 2,4-dichlorphenoxyacetic acid 0.8-1.2mg/L and maltose 80-100g/L；
Division culture medium is with MS for minimal medium, wherein added with 6-benzyl aminoadenine 0.4-0.6mg/L, KT1.3-1.8mg/L, naphthalene acetic acid 0.03-0.06mg/L and maltose 20-40g/L, solidifies with 4-6g/L agar powder.
4. the method for claim 1, it is characterised in that also include after step (3):
The regeneration plant that microspores culture obtains is put in Fructus Hordei Vulgaris mill water culture nutrient solution and cultivates, again screen, select the Fructus Hordei Vulgaris that chlorophyll content is high and/or stem tiller number is many, and collect its seed, wherein, the Fructus Hordei Vulgaris that described chlorophyll content is high and/or stem tiller number is many refer to screened batch Determination of Chlorophyll content and/or tiller number come before 20% Fructus Hordei Vulgaris；The formula of described Fructus Hordei Vulgaris mill water culture nutrient solution is as follows: NH4NO30.2mM、KNO35mM、Ca(NO3)22mM、MgSO42mM、KH2PO40.1mM、Na2SiO30.5mM、NaFe(III)–EDTA0.05mM、H3BO30.01mM、MnCl20.005mM、ZnSO40.005mM、CuSO40.0005mM、Na2MoO30.0001mM, uses front solution ph dilute hydrochloric acid to regulate to 6.0.
5. method as claimed in claim 4, it is characterised in that described screening again is to adopt water planting mode to screen under human controllable's environment, and described human controllable's environment includes: adopt fluorescent tube illumination, intensity of illumination 1500-2500lx in the controlled environment chamber；Keep room temperature daytime 18-22 DEG C, evening 16-22 DEG C, daytime, night temperature fluctuation 1-2 DEG C.
6. the method for claim 1, it is characterised in that adopting following Fructus Hordei Vulgaris mill water culture nutrient solution to carry out water planting, this nutrient solution prescription is as follows: NH4NO30.2mM、KNO35mM、Ca(NO3)22mM、MgSO42mM、KH2PO40.1mM、Na2SiO30.5mM、NaFe(III)–EDTA0.05mM、H3BO30.01mM、MnCl20.005mM、ZnSO40.005mM、CuSO40.0005mM、Na2MoO30.0001mM, uses front solution ph dilute hydrochloric acid to regulate to 6.0；
Water planting in the controlled environment chamber in carry out, adopt fluorescent tube illumination, intensity of illumination 2000lx, keep room temperature daytime 20 DEG C, evening 18 DEG C, temperature fluctuation 1-2 DEG C.
7. the method preparing barley green product, described method includes:
(1) take Fructus Hordei Vulgaris microspore development and be in the small ear in monokaryon early stage and/or mid-term, K cryogenic treatment 2-3 week；
(2) results sporidiole, cultivates sporidiole, and regeneration acquisition is isozygotied doubled haploid；With
(3) isozygoty described in water planting doubled haploid, the Fructus Hordei Vulgaris that for the chlorophyll content of the Fructus Hordei Vulgaris that screening acquisition processes without abovementioned steps relative to the described Fructus Hordei Vulgaris sporidiole of step (1) and obtains and stem tiller number, chlorophyll content improves and/or stem tiller number increases；With
(4) described Fructus Hordei Vulgaris is prepared into barley green product；
Wherein, described step (2) including:
(2a) sterilize described tassel, then clean；
(2b) sporidiole is separated from described tassel；
(2c) processing sporidiole 36-60 hour in room temperature, dark pretreatment fluid, wherein, described pretreatment fluid contains the CaCl of 4-8% mannitol, 1.0-1.5g/L22-(N-morpholine) ethyl sulfonic acid with 0.8-1.2g/L；
(2d) suspend sporidiole with inducing culture, and by inoculation of suspension liquid in inducing culture, callus induction；With
(2e) induction gained callus proceeding to division culture medium, cultivate and obtain regrowth, isozygoty described in namely doubled haploid；
Wherein, in step (2b), when separating sporidiole, test tube adds tassel and extracting solution, exceed the speed limit rotary-cut with high speed disperser, filter, centrifugal filtrate, thus obtaining sporidiole, wherein, described extracting solution contains the CaCl of 4-8% mannitol, 1.0-1.5g/L22-(N-morpholine) ethyl sulfonic acid with 0.8-1.2g/L；
Wherein, described inducing culture is with N6Culture medium is minimal medium, wherein added with 6-Furfurylaminopurine 0.3-0.8mg/L, 2,4-dichlorphenoxyacetic acid 0.5-1.5mg/L and maltose 70-120g/L；
Described division culture medium is with MS for minimal medium, wherein added with 6-benzyl aminoadenine 0.3-0.8mg/L, KT1.0-2.0mg/L, naphthalene acetic acid 0.03-0.08mg/L and maltose 15-50g/L, solidifies with 3-8g/L agar powder.
8. method as claimed in claim 7, it is characterised in that described method also included before step (1):
A () chooses the barley material that a collection of leaf color is dark green, seedling growth is vigorous, tiller is more, the chlorophyll content of 2 leaf 1 heart stages is measured after water planting, start to add up tiller number, plant height and/or basal part of stem girth to tillering stage, filter out chlorophyll content height according to these statistical indicators, tiller number is many, grow rapid kind；With
B () cultivates described kind, it is thus achieved that the sporidiole described in step (1)；With
If c () barley material only has chlorophyll content height, tiller number is many, the objective trait grown in rapidly, then mutually pollinate after pollen maturation hybridization, obtain hybridization F0 for seed, breed this F0 for seed, obtain F1 generation plant, obtain the sporidiole described in step (1) from this F1 generation plant；
Wherein, the kind that described chlorophyll content is high refers to the strain of 20% before being screened batch Determination of Chlorophyll content and coming；The kind that described tiller number is many refer in being screened batch tiller number come before 20% strain；The rapid kind of described growth refer in the being screened batch speed of growth come before 20% strain.
9. method as claimed in claim 7, it is characterised in that
Inducing culture is with N6Culture medium is minimal medium, wherein added with 6-Furfurylaminopurine 0.4-0.6mg/L, 2,4-dichlorphenoxyacetic acid 0.8-1.2mg/L and maltose 80-100g/L；
Division culture medium is with MS for minimal medium, wherein added with 6-benzyl aminoadenine 0.4-0.6mg/L, KT1.3-1.8mg/L, naphthalene acetic acid 0.03-0.06mg/L and maltose 20-40g/L, solidifies with 4-6g/L agar powder.
10. method as claimed in claim 7, it is characterised in that also include after step (3):
The regeneration plant that microspores culture obtains is put in Fructus Hordei Vulgaris mill water culture nutrient solution and cultivates, again screen, select the Fructus Hordei Vulgaris that chlorophyll content is high and/or stem tiller number is many, and collect its seed；Wherein, the Fructus Hordei Vulgaris that described chlorophyll content is high and/or stem tiller number is many refer to screened batch Determination of Chlorophyll content and/or tiller number come before 20% Fructus Hordei Vulgaris；The formula of described Fructus Hordei Vulgaris mill water culture nutrient solution is as follows: NH4NO30.2mM、KNO35mM、Ca(NO3)22mM、MgSO42mM、KH2PO40.1mM、Na2SiO30.5mM、NaFe(III)–EDTA0.05mM、H3BO30.01mM、MnCl20.005mM、ZnSO40.005mM、CuSO40.0005mM、Na2MoO30.0001mM, uses front solution ph dilute hydrochloric acid to regulate to 6.0.
11. method as claimed in claim 10, it is characterized in that, described screening again is to adopt water planting mode to screen under human controllable's environment, and described human controllable's environment includes: adopt fluorescent tube illumination, intensity of illumination 1500-2500lx in the controlled environment chamber；Keep room temperature daytime 18-22 DEG C, evening 16-22 DEG C, daytime, night temperature fluctuation 1-2 DEG C.
12. method as claimed in claim 7, it is characterized in that, described barley green product is selected from: barley fiber food, Folium Hordei Vulgaris urge health food, wheat straw leaf beverage, barley green wheat enzyme nutriment laughable, blue or green, and add the tonic food of dextrin, yeast, Radix Dauci Sativae powder, Korean Ginseng powder in yong barley leaves juice powder.
13. method as claimed in claim 7, it is characterised in that adopting following Fructus Hordei Vulgaris mill water culture nutrient solution to carry out water planting, this nutrient solution prescription is as follows: NH4NO30.2mM、KNO35mM、Ca(NO3)22mM、MgSO42mM、KH2PO40.1mM、Na2SiO30.5mM、NaFe(III)–EDTA0.05mM、H3BO30.01mM、MnCl20.005mM、ZnSO40.005mM、CuSO40.0005mM、Na2MoO30.0001mM, uses front solution ph dilute hydrochloric acid to regulate to 6.0；
Water planting in the controlled environment chamber in carry out, adopt fluorescent tube illumination, intensity of illumination 2000lx, keep room temperature daytime 20 DEG C, evening 18 DEG C, temperature fluctuation 1-2 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310713460.4A CN103636505B (en) | 2013-12-20 | 2013-12-20 | A kind of compound selection of high chlorophyll many tillers Fructus Hordei Vulgaris |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310713460.4A CN103636505B (en) | 2013-12-20 | 2013-12-20 | A kind of compound selection of high chlorophyll many tillers Fructus Hordei Vulgaris |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103636505A CN103636505A (en) | 2014-03-19 |
| CN103636505B true CN103636505B (en) | 2016-06-29 |
Family
ID=50241924
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310713460.4A Active CN103636505B (en) | 2013-12-20 | 2013-12-20 | A kind of compound selection of high chlorophyll many tillers Fructus Hordei Vulgaris |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103636505B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105340755B (en) * | 2015-12-04 | 2018-09-07 | 上海市农业科学院 | Microspore continuously cultivates high-frequency regeneration plant method in cereal crop single plant source |
| CN106106157A (en) * | 2016-06-29 | 2016-11-16 | 无锡南理工科技发展有限公司 | A kind of breeding method of crisp stem Fructus Hordei Vulgaris |
| CN109105258A (en) * | 2018-08-15 | 2019-01-01 | 上海市农业科学院 | A kind of cultural method of barley microspore |
| CN112273219B (en) * | 2020-10-10 | 2022-05-17 | 上海市农业科学院 | A method for binding F by hybridization1Method for rapidly obtaining stable homozygous nitrogen high-efficiency material by culturing microspore |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1418950A (en) * | 2002-12-26 | 2003-05-21 | 北京市海淀区植物组织培养技术实验室 | Method for obtaining haploid plant strain by culturing anther or pollen |
| CN101558735A (en) * | 2009-06-03 | 2009-10-21 | 西北农林科技大学 | Novel method for cultivating wheat stripe rust resistant polygene polymerized flower |
| CN101904296A (en) * | 2010-07-14 | 2010-12-08 | 江苏沿海地区农业科学研究所 | Breeding method of salt-tolerant malting barley variety |
| CN101946629A (en) * | 2010-09-26 | 2011-01-19 | 甘肃省农业科学院生物技术研究所 | Method for rapidly obtaining pure line of hybrid wheat |
| CN102106258A (en) * | 2009-12-24 | 2011-06-29 | 上海市农业科学院 | Method for improving low-nitrogen tolerance character of wheat crops |
| CN102106257A (en) * | 2009-12-24 | 2011-06-29 | 上海市农业科学院 | Method for screening anti-fusarium graminearum toxin of wheat crops |
| CN103249836A (en) * | 2010-09-24 | 2013-08-14 | 巴斯夫植物科学有限公司 | Plants having enhanced yield-elated traits and method for making the same |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013123051A1 (en) * | 2012-02-14 | 2013-08-22 | The Board Of Trustees Of The Leland Stanford Junior University | Method for modulating the number of archesporial cells in a developing anther |
-
2013
- 2013-12-20 CN CN201310713460.4A patent/CN103636505B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1418950A (en) * | 2002-12-26 | 2003-05-21 | 北京市海淀区植物组织培养技术实验室 | Method for obtaining haploid plant strain by culturing anther or pollen |
| CN101558735A (en) * | 2009-06-03 | 2009-10-21 | 西北农林科技大学 | Novel method for cultivating wheat stripe rust resistant polygene polymerized flower |
| CN102106258A (en) * | 2009-12-24 | 2011-06-29 | 上海市农业科学院 | Method for improving low-nitrogen tolerance character of wheat crops |
| CN102106257A (en) * | 2009-12-24 | 2011-06-29 | 上海市农业科学院 | Method for screening anti-fusarium graminearum toxin of wheat crops |
| CN101904296A (en) * | 2010-07-14 | 2010-12-08 | 江苏沿海地区农业科学研究所 | Breeding method of salt-tolerant malting barley variety |
| CN103249836A (en) * | 2010-09-24 | 2013-08-14 | 巴斯夫植物科学有限公司 | Plants having enhanced yield-elated traits and method for making the same |
| CN101946629A (en) * | 2010-09-26 | 2011-01-19 | 甘肃省农业科学院生物技术研究所 | Method for rapidly obtaining pure line of hybrid wheat |
Non-Patent Citations (2)
| Title |
|---|
| Barley anther culture: assessment of carbohydrate effects on embryo yield, green plant production and differential plastid development in relation with albinism;Geneviève Wojnarowiez，et al.;《J.plant physiol.》;20041231;第161卷;第747-755页 * |
| 不同大麦品种的麦绿素专用价值初步研究;王仁杯等;《浙江农业科学》;20031231(第5期);摘要，第256页左栏第2段、右栏第1段，第257页左栏第1-2段及表2 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103636505A (en) | 2014-03-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101120653B (en) | Seedless monordica grosvenori and cultivating method thereof | |
| CN104542256B (en) | A kind of long-grained nonglutinous rice false smut resistance efficiently imports the breeding method of japonica rice | |
| CN104429932B (en) | A kind of Xian round-grained rice hands over the anti-banded sclerotial blight breeding new method of gene introgression | |
| CN103636505B (en) | A kind of compound selection of high chlorophyll many tillers Fructus Hordei Vulgaris | |
| CN107517850A (en) | A kind of haploid method for creating of muskmelon | |
| CN102550405A (en) | Breeding method of poplar haploid | |
| CN102228005A (en) | Pinellia ternate tissue culture one-step speciation method | |
| CN102210259B (en) | Method for achieving isolated embryos through radish and turnip intergeneric distant hybridization | |
| CN1255022C (en) | Paphiopedilum aseptic seeding and tissue culture technology | |
| CN103416304A (en) | Method for cultivating water-saving and drought-resistant rice anther | |
| CN102144560B (en) | Method and application method for obtaining novel germ plasm of brassica A genome vegetable | |
| CN101011028B (en) | Breeding method of chrysanthemum haploid | |
| CN108077061A (en) | A kind of selection made wine with the glutinous two-line sterile lines of Xian | |
| CN103141376B (en) | Cultivation method of rice germplasm with high-resistant starch and low-amylose starch | |
| CN109452166A (en) | A kind of selection of the anti-fall high-quality black rice two-line hybrid rice of elongated grain odor type | |
| CN112442449B (en) | Ramaria original strain culture medium and application thereof as well as Ramaria original strain and culture method thereof | |
| CN108243959A (en) | It is a kind of using yellow fine strain of millet wood stem section as the highly efficient regeneration method of explant | |
| CN101401550B (en) | Method for inducing eggplant sporidiolum to form embryoid and special culture medium thereof | |
| CN104521758B (en) | A kind of Xian Jing hand over gene introgression blast resisting breeding new method | |
| CN114424749B (en) | In-vitro rapid propagation method for liriope spicata | |
| CN108308017B (en) | Cultivation method of virus-resistant interspecific introgression line of sweet potatoes | |
| CN107743868A (en) | A kind of method for efficiently breeding roxburgh anoectochilus terminal bud using nature optical culture forming seedling through one step culture | |
| CN102362579B (en) | Method for cultivating artificial hybridization hexaploid rape microspore regeneration plants | |
| CN105850738B (en) | A kind of eggplant nature Isolated microspore and the haploid method of flower pesticide co-culturing, inducing | |
| CN103798125A (en) | Method for acquiring novel species of brassicaceous vegetables and application method of novel specie of brassicaceous vegetables |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |