CN105309299A - Mechanized hybrid rice seed production using inducible female sterility - Google Patents
Mechanized hybrid rice seed production using inducible female sterility Download PDFInfo
- Publication number
- CN105309299A CN105309299A CN201410360097.7A CN201410360097A CN105309299A CN 105309299 A CN105309299 A CN 105309299A CN 201410360097 A CN201410360097 A CN 201410360097A CN 105309299 A CN105309299 A CN 105309299A
- Authority
- CN
- China
- Prior art keywords
- female
- rice
- gene
- sterile
- paddy rice
- 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.)
- Pending
Links
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention discloses a female sterile rice breeding method. According to the method, an inducible promoter driven rice female fertile gene expression cassette is transformed into female sterile rice; during female sterile line breeding, the transgene rice uses a corresponding induction agent to induce recombinant fertile gene expression so as to achieve fertile selfing breeding; and during hybrid rice seed production, the transgene rice presents completely female sterility when the induction agent is not applied, such that the transgene rice can be adopted as a male parent so as to be used for seed production, and is subjected to hybridization with a female parent to produce hybrid seeds. According to the present invention, during the harvesting of the seed production process, the male parent is female sterility so as not to bear fruits, such that the male parent and the female parent can be subjected to box type sowing or mixed sowing, and the mixed harvesting can be achieved during the harvesting; and the difficult problem of the female sterile rice breeding is solved, the mechanized hybrid rice seed production is achieved, and especially the mechanized hybrid rice seed production in the mixed sowing and mixed harvesting manner is achieved.
Description
Technical field
The invention belongs to agricultural biological technical field, disclose a kind of hybrid rice mechanization producing method for seed utilizing induction type female sterile.
Background technology
The existing hybrid rice seeding technique of China, takes Parent interval sowing, mostly is and first broadcasts male parent, after broadcast female parent; Parent is box separately plants, maternal multirow, male parent duplicate rows; Parent separately harvesting during results, or slice off male parent before pollination after ripening.Therefore, breeding of hybrid rice is a technology that labour intensity is large, operation is loaded down with trivial details, production cost is high.Along with the progressively enforcement of urbanization, industrialization, agriculture industrialization and policy for transferring rural land, the between twenty and fifty labour in stay-at-home rural area successively decreases year by year, and the seeding technique of existing labor-intensive, intensive cultivation type is born without enough labours.Meanwhile, along with the development of national economy and the adjustment of the structure of rural undertaking, farmers' income channel is on the increase, and the production model of this excessive risk of breeding of hybrid rice, low return also lacks attraction to peasant household; And due to part peasant unskilled or think that production of hybrid seeds process is too numerous and diverse, so dare not or be reluctant to bear breeding of hybrid rice work to breeding technology for hybrid rice.Therefore, Mechanicalized seeding method of hybrid rice research is extremely urgent.
Existing many researchers have done various trial and have carried out the hybrid rice mechanization production of hybrid seeds.There are researcher's proposition husk colouring discrimination rice sterile line and restorer seed, specific practice is the rice restorer seed machinery mixed seeding by the rice sterile line seed of normal husk look and improper husk look, through the mixed receipts of machinery after maturation, then use color selector sorting, obtain crossbreed; There is researcher's proposition aircraft to be sprayed by pollen and carry out the production of hybrid seeds in maternal field; Also there is researcher to propose to utilize the larger great disparity of father, maternal grain type, use classificator sorting; Also have the methods such as researcher proposes to use the method for semi-mechanized operation, and namely live female parent, manually plants and gather in male parent, and last mechanical harvesting is maternal; Researcher is also had to utilize paddy rice to carry out the hybrid rice mechanization production of hybrid seeds to the sensitivity characteristic of weed killer herbicide Bentazon.But above-mentioned multiple method all exists many defects and fundamentally can not realize the hybrid rice mechanization production of hybrid seeds, in production, also have no large-scale commercial application at present.
Summary of the invention
The present invention is intended to overcome the deficiencies in the prior art, provides a kind of hybrid rice mechanization producing method for seed utilizing induction type female sterile.
In order to achieve the above object, technical scheme provided by the invention is:
The described hybrid rice mechanization producing method for seed of induction type female sterile that utilizes comprises the steps:
(1) paddy female that inducible promoter drives can be educated gene and insert paddy rice expression vector, paddy rice expression vector is preferably pCAMBIA1300;
(2) the paddy rice expression vector prepared by step (1) is imported female sterile paddy rice, and obtain the transgenic paddy rice isozygotied;
(3) transgenic paddy rice isozygotied is carried out sterile line propagation or the hybrid rice production of hybrid seeds; When carrying out sterile line propagation, apply derivant to the transgenic paddy rice isozygotied, the paddy female that inducible promoter is driven can educate gene expression, and realization can educate self propagated; When carrying out the hybrid rice production of hybrid seeds, under the natural conditions not applying derivant, the transgenic paddy rice isozygotied and maternal male sterile line are planted, hybridization obtains hybrid seed and (shows as complete female sterile when the transgenic paddy rice isozygotied does not apply derivant, thus can be used as male parent and produce hybrid seed for the production of hybrid seeds and hybridization of female parent; During production of hybrid seeds process results, male parent is owing to being female sterile and shaky, and therefore, the no matter box sowing of Parent or mixed seeding, all can mix receipts during results).
Wherein, step (1) described paddy female can educate the wild type gene that gene is the female sterile gene of female sterile paddy rice sfs1, the wild type gene of PTB1 female gene or the wild type gene of FST female sterile gene; The female sterile gene sequence of described female sterile paddy rice sfs1 is as shown in SEQIDNO.1; Described PTB1 female gene sequence is as shown in SEQIDNO.2; Described FST female sterile gene sequence is as shown in SEQIDNO.3.
The DNA sequence dna of step (1) described inducible promoter is as shown in SEQIDNo.4.
The described female sterile paddy rice of step (2) be female sterile paddy rice sfs1 (this male sterile line the number of being applied be 201210096970.7 patent of invention open) or carry PTB1 female gene (this gene the number of being applied be 201010194553.7 patent of invention open) female sterile paddy rice.
Apply derivant in the oogamete puberty to the transgenic paddy rice isozygotied during step (3) described sterile line propagation; The described oogamete puberty is that meiosis stage is to the macrospore puberty.
Described derivant is the composition of propiconazole, nitrile bacterium azoles or propiconazole and nitrile bacterium azoles.
Make use of the transgenic paddy rice comprising above-mentioned expression cassette in the breeding of female sterile paddy rice and the hybrid rice mechanization production of hybrid seeds and carry out the rice material that transformation obtains.
Below in conjunction with principle, the invention will be further described:
First the present invention constructs and carries the paddy rice expression vector that the paddy female driven by inducible promoter can educate the expression cassette of gene.Utilize Transgenic Rice technology, above-mentioned expression vector is proceeded to female sterile paddy rice.During sterile line propagation, above-mentioned transgenic paddy rice utilizes corresponding derivant to induce restructuring fertile gene to express, and makes it educate self propagated; During the hybrid rice production of hybrid seeds, when transgenic paddy rice does not apply derivant, show as complete female sterile, thus can be used as male parent and produce hybrid seed for the production of hybrid seeds and hybridization of female parent.During production of hybrid seeds process results, male parent is owing to being female sterile and shaky, and therefore, Parent can boxly be sowed, also can mixed seeding, can mix receipts during results.
Compared with prior art, beneficial effect of the present invention is:
The inventive method solves a female sterile paddy rice propagation technique difficult problem, achieves the mixed receipts of mixed seeding that hybrid rice seed is produced; During breeding, use chemical inducer, results male parent seed.During the production of hybrid seeds, can boxly broadcast slotting, also can mixed seeding; During results, receipts can be mixed.This mixed seeding mixes the production technology of hybridized rice seed of receipts, can reduce production of hybrid seeds operation sequence, is convenient to carry out mechanized operation, reduces labor intensity, and reduces seed produces cost, the overall benefit improving rice seed industry.
Accompanying drawing explanation
Fig. 1 carries the paddy rice expression vector collection of illustrative plates that the paddy female driven by inducible promoter can educate the expression casette of gene in embodiment 1;
Fig. 2 is PCR detection figure (the M:DNA molecular weight standard of part transfer-gen plant in embodiment 3; 1 ~ 4: transfer-gen plant; 5: negative control)
Fig. 3 be part transfer-gen plant in embodiment 3 Southernblot detection figure (-: negative control; +: using plasmid as positive control);
Fig. 4 is transfer-gen plant pollen I in embodiment 4
2-KI coloration result figure;
Fig. 5 is solid performance (the upper figure: transfer-gen plant does not apply the solid performance after derivant in embodiment 4 after transfer-gen plant applying derivant; Figure below: transfer-gen plant applies the solid performance after derivant);
Fig. 6 be in embodiment 5 paddy female sterile system for reproduction test;
Fig. 7 is embodiment 4 middle and small scale production of hybrid seeds scene photo.
Embodiment
Below by specific embodiment, the invention will be further described, but therefore do not limit the scope of the invention.Involved bacterial strain, carrier, rice varieties are all openly to obtain.
Embodiment 1: the paddy female carrying inducible promoter driving can educate the paddy rice expression vector establishment of the expression casette of gene
Plant expression vector pHB inserts 1 expression cassette: inducible promoter (SEQIDNO.4), wild type PTB1 (paddy female can educate gene) code area (SEQIDNO.2), rbcSPolyA terminator (SEQIDNO.5).Complete expression cassette sequence is as shown in SEQIDNO.6.Accompanying drawing 1 is shown in by complete carrier construction collection of illustrative plates.
Constructed vector plasmid imports agrobacterium strains EHA105 and is used for rice transformation.Introduction method adopts electric shock transformation method, and the electric shock instrument operation instructions of Primary Reference bio-rad company carry out, and concrete steps are as follows:
(1) the DH10B competent cell taking-up being stored in-80 DEG C is placed on freeze thawing on ice, 1mm electric shock cup is placed on precooling on ice, frozen SOC is placed on 37 DEG C and thaws.
(2) clean EP centrifuge tube is inserted in precooling on ice, more than the sample that will transform two of the number of general EP centrifuge tube, one is used for doing negative control (not adding DNA), another does positive control (adding the 10ng/ μ lpUC19 of 1 μ l).
(3) DNA sample that absorption 1-2 μ l will transform respectively puts into the EP centrifuge tube of precooling, then the DH10B competent cell 20 μ l of thawing is taken out gently bottom the EP centrifuge tube putting into precooling, both are mixed gently, try not to produce bubble, do not catch bottom centrifuge tube tactile, avoid variations in temperature to impact competence transformation efficiency, it is fast that operating process is tried one's best.
(4) arrange Transformation Parameters, 1800v, 25 μ F, 200 Ω, 1mm, BioRad electric exciter generally has the operation parameter of recommendation.
(5) gentle aspiration competence and DNA mixture, put into electric shock cup, raps at the bottom of mixture is distributed in glass uniformly, change upper cover and put into electric shock groove, shut safety head, press electric shock red button, after electric shock completes (generally meeting instrument can send prompt tone), electric shock cup is put into by 37 DEG C of preheated SOC, by mixture spin-up, with suction nozzle mixture proceeded to and shake in tube, in 37 DEG C of constant-temperature tables, 225rpm, 1hr.
Embodiment 2: transgenic paddy rice obtains
2.1 vegetable materials are selected
Paddy rice (OryzasativaL.) Fruit variety for genetic transformation is the corresponding paddy female sterile system of carrying PTB1 gene (SEQIDNO.1).
The induction of 2.2 paddy rice embryo callus
After getting pollination, the prematurity rice paddy seed of 10-15d peels off seed coat, in 70% alcohol-pickled 3min, 15min (or in shaking table vibration sterilizing 25min in the liquor natrii hypochloritis proceeding to 20% again) is soaked again in 0.1% mercuric chloride, sterile water wash 3-5 time in superclean bench, the rataria tweezers of seed after sterilization are extruded, be inoculated on inducing culture, each ware about puts 35,28 DEG C of light culture 4-5d, excision radicle, continues to cultivate 12-15d, after callus is grown up, carries out squamous subculture, every two weeks subcultures once, are total to subculture 2-3 time.
Mature embryo shells, and after said method sterilization, is placed on inducing culture.28 DEG C of light culture are to growing callus.Select cultivation or the preculture callus of 4 days as converting material.
The preculture of 2.3 callus
Get eugonic embryo callus (selecting dispersed from squamous subculture, the embryo callus subculture particle of the 2-3mm of color cadmium yellow) and be transferred to precultivation medium, 27 DEG C of light culture 3-4 days.Eugonic callus is used for transforming.
2.4 agriculture bacillus mediated rice conversion
The cultivation of Agrobacterium.Choose the single colony inoculation of Agrobacterium in containing on 50mg/LYM medium, 260C light culture, after 2 ~ 3 days, is collected Agrobacterium thalline with a metal spoon, is suspended in AAM liquid suspension culture base.Adjustment bacterium colony concentration is 0.8 ~ 1.0 to O.D.600, namely can be used for transforming.
The Dual culture of callus and Agrobacterium.All should first preculture 3 ~ 4 days on fresh subculture medium IM before being polluted by Agrobacterium for the callus that transforms.To transfer to through pre-incubated callus in an aseptic triangular flask during conversion, then pour appropriate agrobacterium suspension (at least ensuring enough bacterium liquid and material) into, ambient temperatare puts 10 ~ 20 minutes, and frequently rocks.Take out callus, aseptic paper suck unnecessary bacterium liquid, with transfer to by callus be covered with one deck aseptic filter paper solidified co-cultivation medium CM in 260C light culture 2 ~ 3 days.
The screening of 2.5 kanamycin-resistant callus tissues
Callus is transferred on Selective agar medium and screen kanamycin-resistant callus tissue, switching in every two weeks 1 time.Cultivate 4-8 week, most callus brownization is dead, has minority warty kanamycin-resistant callus tissue to grow from the callus surface of shrivelled brownization.Select these kanamycin-resistant callus tissues and continue light culture 2 times on Selective agar medium, select a part for callus after growing up and transfer on differential medium.
The differentiation of 2.6 kanamycin-resistant callus tissues is cultivated
Through the kanamycin-resistant callus tissue that antibiotic-screening grows, proceed in differential medium and continue to cultivate, 26 DEG C ~ 28 DEG C, 12h illumination, cultivates under 12h dark condition.
The regeneration of 2.7 transfer-gen plants and seedling replanting
Transfer to the callus on differential medium, cultivate callus after 2 weeks and start to turn green, can put out new shoots after 3 weeks, root also grows thereupon.Seedling is transferred in the little triangular flask containing root media, every bottle of strain, continue illumination cultivation, when height of seedling 7 ~ 10cm, open bottle cap hardening 5 ~ 7 days in greenhouse, after seedling robust growth, shift out blake bottle, clean the medium of catching up with, move to greenhouse pot culture (in iron pan).Note moisturizing, to improve transplanting survival rate.
Embodiment 3: the Molecular Detection of transfer-gen plant
The extensive extracting of 3.1DNA
T
0cTAB method (MurryandThompson, 1980) is adopted for transgenic paddy rice DNA extracting.Take fresh blade 2-4 gram, put into the mortar of-20 DEG C of precoolings after shredding, clay into power with liquid nitrogen, proceed in the grind away bottle of precooling and save backup in-20 DEG C.Add the 10ml1.5XCTAB being preheated to 100 DEG C during extraction, stir evenly rapidly, vibrate 20 minutes in 56 DEG C of water bath heat preservation shaking tables, then use equal-volume chloroform: isoamyl alcohol (24:1) extracting.After centrifugal, supernatant is preheated to the 10%CTAB buffer solution of 56 DEG C and isopyknic chloroform with 1/10 volume again: isoamyl alcohol (24:1) extracting once, then supernatant adds 1%CTAB precipitation DNA, the 1mol/LNaCl dissolving DNA being added with RNaseA is added after centrifugal, spend the night in 56 DEG C and dissolve after alcohol precipitation completely, be dissolved in appropriate TE, measure DNA concentration with DNA microdetermination instrument (DNAFluorometer), balance DNA concentration to 250ng/ul, be stored in 4 DEG C for subsequent use.
3.2PCR analyze
For transgenic paddy rice DNA, PCR detection is carried out to T0.Using promotor as template design primer.Forward primer is: 5 '-cgccaattgaagctttaaattatttttccaatatttattttttta-3 ' (SEQIDNO.7), and reverse primer is: 5 '-tatggatccgtcgacactagtccgatcggtgatccctcctc-3 ' (SEQIDNO.8).Amplified production fragment is 2797bp.PCR reaction system: DNA30-90ng, 10xBuffer2.0ul, 1mMdNTP1.8ul, 25mMMgCl
2each 0.5ul, the Tag enzyme 1.5U of 1.5ul, 10uMprimer two kinds, then adds ddh
2o to 20ul reaction volume.PCR cycling condition: 94 DEG C, sex change 3 minutes; 94 DEG C, 1 minute, 55 DEG C, 1.5 minutes, 72 DEG C, 1.5 minutes, 40 circulations; 72 DEG C, extend 5 minutes.Electrophoresis detection: use 1.4% agarose gel electrophoresis, film recording electrophoresis result.
Accompanying drawing 2 is partial transgenic plant pcr amplification result, shows that genes of interest has been integrated in rice genome.
3.3Southern engram analysis
Get rice total dna 2.5ug, with restriction enzyme XbaI37 DEG C of enzymolysis about 20hr, cut entirely through electrophoresis detection enzyme, with 0.6 ~ 0.8% agarose (production of Sigma company) gel electrophoresis, after 12-16hr electrophoresis, gel 0.2NHCl sex change 10 minutes, is transferred to nylon membrane Hybond-N with 0.4NNaOH solution by the DNA fragmentation on gel
+on.Transferase 12 0 hour, by nylon membrane 2XSSC rinsing 2 times, each 5 minutes, in 80-100 DEG C of vacuum bakeout 3hr after film dries, taking-up cooling was placed in 4 DEG C of refrigerators and saves backup.
Prehybridization: first nylon membrane is used 2XSSC rinsing, loads in hybridization bag after airing, add the hybridization buffer that 10-18ml (the number 1-6 according to film opens) is preheated to 65 DEG C.After catching up with bubble, prehybridization about 6hr in 65 DEG C of air tables.
Probe mark: follow these steps to mark with random priming.DNA probe 100ng, adds ddH
2o to 10ul, 100 DEG C of sex change 10 minutes, after ice bath cooling in 5 minutes, add dNTP (A+G+T) 3ul, random primer buffer solution 5.0ul, KlenowFragment enzyme 2U, finally adds a-
32p-dCTP10-40uCi (by hybond membrane quantity), in 25 DEG C of insulations more than 3 hours after mixing.
Hybridization: add 600ul hybridization solution in the probe solution marked, open pipe lid, 100 DEG C of sex change, after 10 minutes, are poured in hybridization bag, 65 DEG C of hybridization 6hr.
Wash film, compressing tablet: after hybridization, film is taken out from hybridization bag, with film washing liquid 1XSSC, 0.2%SDS room temperature rinsing 1 time, and then wash 1-2 time by the film washing liquid heat in 65 DEG C of insulation shaking tables being preheated to 65 DEG C, each 15 minutes.Use preservative film coating after airing, after compressing tablet, be about 4-7 days in-20 DEG C of autoradiograph.
Prepared by DNA probe.DNA probe is selected from the pcr amplification product in 3.2, and the sequence of amplified production is shown in SEQIDNO.4.
Accompanying drawing 3 is the Southern engram analysis of partial transgenic plant.Result shows, genes of interest has been integrated in rice genome.
Embodiment 4: the fertility of transgenic rice plant is investigated
Pollen fertility and solid situation have been investigated to the transfer-gen plant through Molecular Identification.Flower pesticide when taking away colored, through the I of routine
2after-KI dyeing, observe the fertile flower powder (see accompanying drawing 4) that whole pollen grain is black dye; Compare with female sterile, transfer-gen plant can be shaky, shows as complete female sterile (see accompanying drawing 5).
Embodiment 5: paddy female sterile system is used for reproduction test
The breeding of paddy female sterile system.The paddy female sterile system selfing of homozygous transgenic will be carried.Selfed seed, by the photometric sorting to red fluorescence, can filter out the seed with red fluorescence, obtains the female sterile paddy rice of isozygotying the mixes receipts hybrid rice mechanization production of hybrid seeds for mixed seeding; And carry genetically modified heterozygosis seed, continue on for the breeding of female sterile paddy rice.
In flower pesticide meiosis stage of transgenic paddy rice to the little armful of sub-phase of monokaryon, blade is sprayed application to 0.05cy0 third ring mile or 0.05cy0 eyeball bacterium mile or 0.05% the third ring mile+0.05% eyeball bacterium mile mixed aqueous solutions, after 3 days, heavy dressing once, and " result shows; the self-fruitful rate being used alone the third ring mile or eyeball bacterium mile is about 60%, and the self-fruitful rate that 2 kinds of derivants are mixed is about 86% (Fig. 6)
Embodiment 6: paddy female sterile system is used for production of hybrid seeds test
Paddy female sterile system is applied to the hybrid rice production of hybrid seeds.Because male parent (restorer) is Female sterile clone, during the production of hybrid seeds, do not use derivant, therefore can boxly plant during the production of hybrid seeds, also can the mixed receipts of mixed seeding.Parent presses the quantitative proportion Homogeneous phase mixing of 2:10 ~ 20, once sows, and can carry out seedling and field production management as conventional rice, can live, rice transplanting, the machine transplanting of rice etc.After maturation, carry out mechanized harvest, results be hybrid rice seeds.It is on-the-spot that accompanying drawing 7 illustrates the small-scale production of hybrid seeds.
Claims (6)
1. utilize a hybrid rice mechanization producing method for seed for induction type female sterile, it is characterized in that, described method comprises the steps:
(1) paddy female that inducible promoter drives can be educated gene and insert paddy rice expression vector;
(2) the paddy rice expression vector prepared by step (1) is imported female sterile paddy rice, and obtain the transgenic paddy rice isozygotied;
(3) transgenic paddy rice isozygotied is carried out sterile line propagation or the hybrid rice production of hybrid seeds; When carrying out sterile line propagation, apply derivant to the transgenic paddy rice isozygotied, the paddy female that inducible promoter is driven can educate gene expression, and realization can educate self propagated; When carrying out the hybrid rice production of hybrid seeds, the transgenic paddy rice isozygotied and maternal male sterile line are planted under the natural conditions not applying derivant, hybridization obtains hybrid seed.
2. the method for claim 1, it is characterized in that, step (1) described paddy female can educate the wild type gene that gene is the female sterile gene of female sterile paddy rice sfs1, the wild type gene of PTB1 female gene or the wild type gene of FST female sterile gene; The female sterile gene sequence of described female sterile paddy rice sfs1 is as shown in SEQIDNO.1; Described PTB1 female gene sequence is as shown in SEQIDNO.2; Described FST female sterile gene sequence is as shown in SEQIDNO.3.
3. the method for claim 1, is characterized in that, the DNA sequence dna of step (1) described inducible promoter is as shown in SEQIDNo.4.
4. the method for claim 1, is characterized in that, the described female sterile paddy rice of step (2) is female sterile paddy rice sfs1 or the female sterile paddy rice of carrying PTB1 female gene.
5. the method for claim 1, the phase is characterised in that, is to apply derivant in the oogamete puberty to the transgenic paddy rice isozygotied during step (3) described sterile line propagation; The described oogamete puberty is that meiosis stage is to the macrospore puberty.
6. method as claimed in claim 5, it is characterized in that, described derivant is the composition of propiconazole, nitrile bacterium azoles or propiconazole and nitrile bacterium azoles.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410360097.7A CN105309299A (en) | 2014-07-25 | 2014-07-25 | Mechanized hybrid rice seed production using inducible female sterility |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410360097.7A CN105309299A (en) | 2014-07-25 | 2014-07-25 | Mechanized hybrid rice seed production using inducible female sterility |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105309299A true CN105309299A (en) | 2016-02-10 |
Family
ID=55238403
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410360097.7A Pending CN105309299A (en) | 2014-07-25 | 2014-07-25 | Mechanized hybrid rice seed production using inducible female sterility |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105309299A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109486853A (en) * | 2018-11-21 | 2019-03-19 | 湖南杂交水稻研究中心 | A method of quickly formulating the engineering Female sterile clone for being suitble to the mechanization production of hybrid seeds using genome editing technique |
| CN114317599A (en) * | 2021-12-29 | 2022-04-12 | 云南农业大学 | Rice female nuclear sterility genetic engineering expression vector and application thereof |
-
2014
- 2014-07-25 CN CN201410360097.7A patent/CN105309299A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109486853A (en) * | 2018-11-21 | 2019-03-19 | 湖南杂交水稻研究中心 | A method of quickly formulating the engineering Female sterile clone for being suitble to the mechanization production of hybrid seeds using genome editing technique |
| CN114317599A (en) * | 2021-12-29 | 2022-04-12 | 云南农业大学 | Rice female nuclear sterility genetic engineering expression vector and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103834684B (en) | Method for mechanically producing seed by using female sterile hybrid rice | |
| CN101810140B (en) | Method for propagating dendrobium candidum test-tube plantlets | |
| CN100557014C (en) | A kind of nodule azotobacter strain is BXYD3 and application thereof | |
| CN107460133B (en) | Dark color has every endogenetic fungus HS40 and its application in dendrobium candidum production | |
| CN104004781A (en) | Preparation method of glyphosate resistant transgenic rice | |
| CN109486853A (en) | A method of quickly formulating the engineering Female sterile clone for being suitble to the mechanization production of hybrid seeds using genome editing technique | |
| CN107125005B (en) | Mycorrhizal seedling raising method for paphiopedilum harderi | |
| CN100557015C (en) | A kind of nodule azotobacter strain is BXBL9 and application thereof | |
| CN111793567A (en) | Mucoraceae fungus and application thereof in promoting paphiopedilum brandisil seeds to germinate and form seedlings | |
| CN101928724A (en) | Mechanical hybrid rice seed production method utilizing transgenic technology of chloroplasts | |
| CN111019835A (en) | Method for separating Armillaria mellea from ambary endophytic fungi | |
| CN105309299A (en) | Mechanized hybrid rice seed production using inducible female sterility | |
| EP3247787A1 (en) | New hybrid agaricus bisporus mushrooms strains | |
| CN110305894B (en) | Rapid and efficient catalpa bungei genetic transformation method | |
| CN101748073B (en) | Method for separating and preserving Cordyceps militaris spawn | |
| CN104620983B (en) | A kind of barnyard grass tissue is cultivated the method for seedling | |
| CN101948868B (en) | Genetic transformation method taking sweet sorghum young ear or young ear induced callus as explant | |
| CN110073898A (en) | A kind of ladder rib hickory chick strain the factorial production and northern facility cultivation technique | |
| CN113796260B (en) | Poria (Wolfiporia cocos) YX1, and culture medium and cultivation method thereof | |
| CN114946524A (en) | Efficient cultivation method for morchella | |
| CN106613970B (en) | The quick breeding by group culture method of sealwort leaf elegant jessamine | |
| CN112961787B (en) | Agrocybe aegerita strain and cultivation method thereof | |
| CN115975817A (en) | Ganoderma lucidum strain L5478 and artificial cultivation method thereof | |
| CN104946644A (en) | Molecular marker for corn tripsacum monosome addition line nucleic male sterility genes and application thereof | |
| CN101182478B (en) | Root nodule azotobacter strain BDYD1 and uses thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160210 |
|
| WD01 | Invention patent application deemed withdrawn after publication |