CN104542260A - Alfalfa cytoplasm male sterility line cms4-10 germplasm and method for breeding and propagating germplasm - Google Patents
Alfalfa cytoplasm male sterility line cms4-10 germplasm and method for breeding and propagating germplasm Download PDFInfo
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Abstract
The invention relates to an alfalfa cytoplasm male sterility line cms4-10 germplasm. The germplasm is prepared by adopting a method comprising the following steps: (1) taking alfalfa cultivation varieties after hole planting for 3 years, namely Gannong No.3, Tianshui, Muge Graze401+Z, Golden Empress, Congnong No.1, Qingyang, Hexi, Gannong No.1, Western Giants, Longzhong, Jindalai, Gannong No.2, Giant, Algonquin, Alfaking, Dingxi, Longdong and Zhonglan No.1 as a natural male sterility mutant screening germplasm resource library; (2) regularly grading and measuring pollen quantity of each alfalfa variety; (3) preparing an Alexander staining solution; (4) measuring pollen fertility rate of a single plate; (5) judging whether the pollen fertility is male fertility or female fertility; and (6) screening the natural male sterility mutant screening germplasm resource library in the step (1) to obtain male sterile plants cms4-10 of the alfalfa variety Muge Graze 401+Z. The invention also discloses a method for breeding and propagating the germplasm. The alfalfa cytoplasm male sterility line cms4-10 germplasm is stable in characteristics, clear in breeding objective and simple in breeding process.
Description
Technical field
The present invention relates to leguminous forage and forage crop Biotechnology in Genetic Breeding field, particularly relate to alfalfa cells matter male sterile line cms4-10 kind matter and seed selection thereof and propagation method.
Background technology
Alfalfa (
medicago satival.) be perennial autotetraploid and cross-pollinatd plant, germ plasm resource is enriched, kind is many and distribution is wide, between abundant alfalfa kind matter, normal producer exchanges and plants matter and gradually oozes, make its hereditary basis show sizable heterogeneity (Wang Xiaojuan etc. the colony's tracer method based on RAPD analyzes Alfalfa Germplasms diversity. plant research. 2010. 30:87-91.).Simultaneously, due to alfalfa cross pollination and hand over mutually can be pregnant, thus create a large amount of genetic variation, this be the screening of natural malesterile mutants material in alfalfa breeding work provide advantage (Li Xiaoxia etc. several alfalfa Asia (change) plants pollen and ovule Genetic Variation Analysis. Scientia Agricultura Sinica. 2009. 42:1911-1917.).Chinese scholars lasted for more than 40 years to the research of alfalfa male sterile line, some clover male sterile lines are obtained, but because the male sterile line Pistil Fertility obtained is low, hybrid seeding ripening rate is low, production is difficult to large scale application, and the alfalfa male sterile line that therefore seed selection is excellent has become an important process of breeding scholar.
At present, the pollen control system of domestic and international alfalfa heterosis utilization mainly comprises following several types: cytoplasmic male sterility (cytoplasmic male sterility, CMS), nuclear male sterility (genetic male sterility, GMS), nucleo-cytoplasmic interreaction male sterility (cytoplasmic genetic male sterility, CGMS), self-incompatibillity (self-incompatibility, and Transgenic male sterile (transgenic male sterility) SI), wherein, cytoplasmic male sterility be alfalfa heterosis utilization important channel (Feng Guanghui etc. alfalfa male sterile mechanism and hybrid seeding progress. seed. 2011. 30:55-59.).Cytoplasmic male sterility is because of the characteristic of its maternal inheritance, and sterile proterties can pass to filial generation by the genetic stability of parental cells plasmagene, can find maintainer, provides foundation for producing the heterotic utilization of upper male sterile.
1967, Davis & Greenblatt(Davis WH, Greenblatt IM. Cytoplasmic male sterility in alfalfa. Journal of Heredity. 1967. 58:301-305.) from multiple alfalfa variety, filter out holandry sterile plant, also repeatedly backcross after hybridizing with fertile plant, filial generation all shows male sterile, there is maternal inheritance characteristics, belong to cytoplasmic inheritance, this is also report guided cell matter male sterile phenomenon in alfalfa first.Bradner & Childers(Bradner NR subsequently, Childers WR. Cytoplasmic male sterility in alfalfa. Canadian Journal of Plant Science. 1968. 48:111-112.) find in alfalfa variety Ontario pollen abortion rate up to 95% sterile plant, and called after Oms-5, genetic test confirms that this sterile strain belongs to cytoplasmic male sterility type.Stucker & Pedersen(Pedersen MW, Stucker RE. Evidence of cytoplasmic male sterility in alfalfa. Crop Science. 1969. 9:767-770.) also in succession find in alfalfa variety DuPuits and reported Somaclonal variation body.Goldberg(Goldberg RB. Plants:novel developmental processes. Science. 1988. 240:1460-1467.) carry out the male sterile genetic transformation of alfalfa, ribalgilase (Barnase) gene with tapetal promoter pTA29 is proceeded to clover and obtains transfer-gen plant.(the Rosellini D such as Rosellini, Ferranti F, Barone P, Veronesiet F. Characterization of transgenic male sterility in alfalfa. Euphytica. 2001. 118:313-319.) find when further the hereditary capacity of the sterile strain of this Transgenic male being analyzed, compared with normal alfalfa plants, the male sterile plant meiotic stage pollen turning pTA29-barnase is less, there is the thinning and cell evacuation phenomenon of tapetum, and cytoplasm there will be degraded in various degree, this directly results in little Hua and flicks the reduction with pollen germination rate too early.But these achievements in research are only only limitted to the Study on Resources stage, also differ greatly from practical application.
The primary breeding objective of Alfalfa Breeding worker obtains to have heterotic crossbreed to improve the yield and quality of herbage, and wherein alfalfa crossbreed production system mainly concentrates on and utilizes cytoplasmic male sterile line to carry out hybrid seeding.Up to now, many countries such as the U.S., Canada, the former Soviet Union have carried out the seed selection of alfalfa cells matter male sterile line, hybrid experiment data show to utilize the male sterile of alfalfa cells matter to hybridize the crossbreed obtained, volume increase 10% ~ 50% is not etc., there is obvious hybrid vigour and considerable production application value, and the sterile cytoplasm of these alfalfa cells matter sterile hybrids all derives from alfalfa cultivar.Between 2001 ~ 2006 years, Dairyland seeds company of the U.S. was through the effort of twenties years, achieve commercially producing of alfalfa sterile hybrid, the alfalfa crossbreed selected except volume increase, also have resistance toly to cradle, grow the characteristic such as fast (Sun Huan etc. pigeonpea, clover and soybean 3 kinds of legume heterosis utilizations general introductions. Scientia Agricultura Sinica. 2009. 42 (5): 1528-1539.).At present, the domestic patented technology also do not had about alfalfa cells matter male sterile line kind matter and seed selection and breeding, only relate to the method being carried out seed selection clover male sterile line by distant hybridization, as " selection of clover male sterile line " that the patent No. is CN201010624142.7, the method with the Wild Medicago Falcata on Xilinguole Beit grassland for female parent, with the accurate Ge Er alfalfa in the Inner Mongol for male parent carries out interspecific hybridization, male sterile line Ms-4 is obtained from its offspring, this male sterile line and 9 alfalfa cultivars carry out test cross, find F
12 ~ 10% are accounted for, the male sterile type that genus anas gene controls for male sterile plants in colony.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of alfalfa cells matter male sterile line cms4-10 kind matter.
Another technical problem to be solved by this invention is to provide seed selection and the propagation method of simple this alfalfa cells matter male sterile line cms4-10 kind matter of with clearly defined objective, program.
For solving the problem, alfalfa cells matter male sterile line cms4-10 kind matter of the present invention, is characterized in that: this kind of matter adopts following method to obtain:
Using bunch planting plant 3 years No. 3, the sweet agriculture of alfalfa cultivar, the sky and water, pastoral song Graze401+Z, golden queen, No. 1, public agriculture, Qingyang, the west of a river, No. 1, sweet agriculture, western giant, in Gansu Province, Jin Dalai, No. 2, sweet agriculture, giant, Algonquin, clover king, Dingxi, Long Dong and middle blue No. 1 as natural malesterile mutants screening germplasm resource bank;
(2) each alfalfa cultivars is in florescence Stochastic choice 10 strain, random selecting 5 little Hua on the different bifurcation level of every strain, play little Hua pollen on the black plastic plate of neat and tidy, adopt pollen amount fractionation testing method according to the level Four classification without pollen, a small amount of pollen, appropriate pollen, pollinia according to a conventional method, estimate pollen amount rank and record;
(3) prepare Alexander dyeing liquor:
The malachite green of I preparation 1%: the malachite of the 1g alcohol that the mass concentration of 100 ml is 95% is dissolved, and stirs 10 min, then leave standstill 3 h and get final product;
II prepares 1% acid fuchsin: dissolved by the acid fuchsin of 1 g 100 ml distilled water, to obtain final product;
III prepares 1% orange red G: dissolved by the orange red distilled water of 1 g, to obtain final product;
10 ml mass concentrations are the ethanol of 95% by V successively, the malachite green of 1 ml 1%, 50 ml distilled water, 25 ml glycerine, 5 g phenol, the acid fuchsin of 5 ml 1%, after the orange red G of 0.5 ml 1% mixes, obtain Alexander dyeing liquor;
(4) individual plant pollen can the rate of educating be measured:
Drawing 150 μ l Alexander dyeing liquors is placed on clean slide, again from 3 inflorescences of random selecting each individual plant, each inflorescence gets 2 little Hua, eject pollen respectively in Alexander dye liquor, dye 30 min, examine under a microscope pollen staining situation, if pollen can be dyed redness by dye liquor, then represent that pollen can be educated; If cyanic colours dyed by pollen envelop, then represent pollen sterility; Choose 5 visuals field, total number of fertile pollen and pollen sterile in the statistics visual field, and be calculated as follows pollen and can educate rate:
FPR=(fertile pollen sum/observe pollen frequence) × 100%;
(5) according to pollen amount classification and pollen staining rate result, if individual plant pollen amount is a large amount of or appropriate, and FPR >=10%, be expressed as male-fertile; Individual plant pollen amount be a small amount of, without pollen, or FPR<10%, be then expressed as male sterile;
To described step (1) in natural malesterile mutants screening germplasm resource bank screen, obtain the male sterile plant 4-10 of alfalfa cultivars pastoral song Graze401+Z.
The seed selection of alfalfa cells matter male sterile line cms4-10 kind matter as above and propagation method, comprise the following steps:
1. using the male sterile plant 4-10 of alfalfa cultivars pastoral song Graze401+Z as female parent, so that the western giant 10-10 of alfalfa can be educated for male parent, conventionally hybridize under chamber planting condition, obtain first generation of hybrid F
1seed;
2. described F is sowed
1seed, identifies its pollen quantity and Pollen Activity in the florescence, whole F
1it is sterile that plant is holandry, and using male sterile individual plant 4-10-2,4-10-4,4-10-5,4-10-6 and 4-10-12 wherein as female parent, and can educate the western giant 10-10 of parent's alfalfa and backcross, and obtains the BCF that backcrosses
1seed;
3. backcross described in sowing BCF
1seed, identifies its pollen quantity and Pollen Activity in the florescence, whole BCF
1it is sterile that plant is holandry, and the male sterile plant 4-10 that name obtains is cms4-10, its F
1male sterile plant 4-10-2,4-10-4,4-10-5,4-10-6 and 4-10-12 as cms4-10 Derivative line, called after cms4-10-2, cms4-10-4, cms4-10-5, cms4-10-6 and cms4-10-12;
4. before autumn, alfalfa was gathered in the crops, select and remain male sterile line cms4-10 as maternal plant according to parent's standard, alfalfa western giant 10-10 maternal plant is as paternal plant, according to a conventional method cuttage clonal propagation is carried out to described maternal plant and described paternal plant, safe overwintering, in field planting at the beginning of 3 months spring of next year in 40 ~ 60 order net isolation method districts, described maternal plant and described paternal plant are in the ratio field planting of 4 ~ 5:1, plantation density is line-spacing 60 cm, spacing in the rows 40 cm, honeybee is put at net indoors artificial in the florescence, maternal plant is authorized by paternal plant pollen, paternal plant is pulled out after blossom fall, seed was gathered in the brown pod phase.
The present invention compared with prior art has the following advantages:
1, the present invention overcomes the defect of existing alfalfa seeding technique, with the excellent cultivar of autotetraploid alfalfa for germplasm resource bank, separates nature male-sterile mutation strain 4-10 from alfalfa cultivars pastoral song, by interspecific cross and the acquisition F that backcrosses
1generation and BCF
1for seed, detect screening through pollen fertility and economical character and obtain sterility 100% genetic stability and the excellent alfalfa cells matter male sterile line cms4-10 of economical character and Derivative line cms4-10-2, cms4-10-4, cms4-10-5, cms4-10-6 and cms4-10-12, provide cytoplasmic male sterility new germ plasm for the heterosis utilization of leguminous forage alfalfa and crossbreed produce.
2, the floral organ of the alfalfa cells matter male sterile line cms4-10 of gained of the present invention is characterized as: the inflorescence properties of alfalfa cells matter male sterile line cms4-10 is similar to maintainer, every inflorescence little Hua number 8-15 piece, diadelphous stamen 10 pieces (9+1), flower pesticide is shrivelled in faint yellow, stamen and anther length normal, examine under a microscope, this cytoplasmic male sterile line is without pollen or pollen debility (see Fig. 1), namely the male sterile of alfalfa cells matter is non-pollen type, insensitive to environment reaction, sterile proterties 100% genetic stability; Gynoecium Dan Sheng, the many mastoid processes of stigma surface, ovary mode of appearance size is identical with fertile line, and 26S Proteasome Structure and Function is normal; Nectary quantity is identical with maintainer, and development condition is good, can attract the normal gathering honey pollination of the insects such as honeybee; Comprehensive agronomy proterties is excellent, dark green leaf color, and stem is red, and growth is neat, and comprehensive disease resistance is strong, drough-resistant and saline-alkali resistant.
Through in Gansu, China province Lanzhou, Zhangye Linze county and the plantation from generation to generation of continuous 5 years 5 of Mongolian gazelle town, Wuwei, its sterile proterties is stablized, and all shows as holandry sterile.
3, breeding objective of the present invention clear and definite (see Fig. 2), the procedure of breeding is simple, and breeding research and utilization are conveniently.
4, alfalfa cells matter male sterile line economical character of the present invention is excellent, and Cross fertile rate is high, is conducive to breeding male sterile lines seed and produces crossbreed (see Fig. 3).
5, compared with alfalfa cells genic male sterile line, the sterile proterties of this cytoplasmic male sterility strain entails filial generation completely by plasmon, and artificialpollination shows the F that cms4-10 and maintainer 10-10 hybridization obtains
1the sterile proterties of holandry is all shown as plant, and F
1normally solid for plant, be excellent alfalfa cells matter male sterile line (see table 1), be suitable for the production application of commercialization crossbreed.
Table 1 cytoplasmic male sterile line cms4-10 and Derivative line thereof and maintainer 10-10 hybridize F
1and BC
1f
1ripening rate
Based on this, the present invention proposes alfalfa cells matter male sterile line (cms4-10 and Derivative line cms4-10-2, cms4-10-4, cms4-10-5, cms4-10-6 and cms4-10-12) hybrid seeding system (see Fig. 4), namely directly utilize male sterile line and there is heterotic strong excellent paternal hybrid and obtain hybrid seed, because alfalfa belongs to the herbage and forage crop of perennial cauline leaf results, male sterile line and maintainer hybridization is directly utilized to obtain hybrid vigour, do not need restorer, compared with genie male sterile line hybrid seeding system, F can be saved
1for seed produces process, substantially reduce the breeding time limit, there is higher economic worth.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 is that the pollen fertility of cms4-10 of the present invention and maintainer 10-10 compares.Wherein A is the dyeing of maintainer 10-10 plant flower pesticide; B is pollen staining after maintainer 10-10 plant anther dehiscence; C is maintainer 10-10 pollen morphology; D is the dyeing of cms4-10 plant flower pesticide; E is pollen staining after cms4-10 plant anther dehiscence; F is cms4-10 plant pollen form.
Fig. 2 is the seed selection flow process of alfalfa cells matter male sterile line cms4-10 of the present invention.
Fig. 3 is that alfalfa cells matter male sterile line cms4-10 of the present invention and maintainer growth traits thereof compare.Wherein A is maintainer 10-10 plant forms; B is cms4-10 plant forms; C is maintainer 10-10 plant inflorescence; D is cms4-10 plant column cap neutral red staining; E is cms4-10 plant inflorescence; F is the pod that cms4-10 plant little Hua is formed.
Fig. 4 is alfalfa cells matter male sterile line cms4-10 heterosis utilization approach of the present invention.
Embodiment
embodiment 1alfalfa cells matter male sterile line cms4-10 kind matter, this kind of matter adopts following method to obtain:
Using bunch planting plant 3 years No. 3, the sweet agriculture of alfalfa cultivar, the sky and water, pastoral song Graze401+Z, golden queen, No. 1, public agriculture, Qingyang, the west of a river, No. 1, sweet agriculture, western giant, in Gansu Province, Jin Dalai, No. 2, sweet agriculture, giant, Algonquin, clover king, Dingxi, Long Dong and middle blue No. 1 as natural malesterile mutants screening germplasm resource bank;
(2) each alfalfa cultivars is in florescence Stochastic choice 10 strain, random selecting 5 little Hua on the different bifurcation level of every strain, play little Hua pollen on the black plastic plate of neat and tidy, adopt pollen amount fractionation testing method according to the level Four classification without pollen, a small amount of pollen, appropriate pollen, pollinia according to a conventional method, estimate pollen amount rank and record;
(3) prepare Alexander dyeing liquor:
The malachite green of I preparation 1%: the malachite of the 1g alcohol that the mass concentration of 100 ml is 95% is dissolved, and stirs 10 min, then leave standstill 3 h and get final product;
II prepares 1% acid fuchsin: dissolved by the acid fuchsin of 1 g 100 ml distilled water, to obtain final product;
III prepares 1% orange red G: dissolved by the orange red distilled water of 1 g, to obtain final product;
10 ml mass concentrations are the ethanol of 95% by V successively, the malachite green of 1 ml 1%, 50 ml distilled water, 25 ml glycerine, 5 g phenol, the acid fuchsin of 5 ml 1%, after the orange red G of 0.5 ml 1% mixes, obtain Alexander dyeing liquor;
(4) individual plant pollen can the rate of educating be measured:
Drawing 150 μ l Alexander dyeing liquors is placed on clean slide, again from 3 inflorescences of random selecting each individual plant, each inflorescence gets 2 little Hua, eject pollen respectively in Alexander dye liquor, dye 30 min, examine under a microscope pollen staining situation, if pollen can be dyed redness by dye liquor, then represent that pollen can be educated; If cyanic colours dyed by pollen envelop, then represent pollen sterility; Choose 5 visuals field, total number of fertile pollen and pollen sterile in the statistics visual field, and be calculated as follows pollen and can educate rate:
FPR=(fertile pollen sum/observe pollen frequence) × 100%;
(5) according to pollen amount classification and pollen staining rate result, if individual plant pollen amount is a large amount of or appropriate, and FPR >=10%, be expressed as male-fertile; Individual plant pollen amount be a small amount of, without pollen, or FPR<10%, be then expressed as male sterile;
To step (1) in natural malesterile mutants screening germplasm resource bank screen, obtain the male sterile plant 4-10 of alfalfa cultivars pastoral song Graze401+Z.
embodiment 2the seed selection of alfalfa cells matter male sterile line cms4-10 kind matter and propagation method, comprise the following steps:
1. using the male sterile plant 4-10 of alfalfa cultivars pastoral song Graze401+Z as female parent, so that the western giant 10-10 of alfalfa can be educated for male parent, conventionally hybridize under chamber planting condition, obtain first generation of hybrid F
1seed;
2. F is sowed
1seed, identifies its pollen quantity and Pollen Activity in the florescence, whole F
1it is sterile that plant is holandry, and using male sterile individual plant 4-10-2,4-10-4,4-10-5,4-10-6 and 4-10-12 wherein as female parent, and can educate the western giant 10-10 of parent's alfalfa and backcross, and obtains the BCF that backcrosses
1seed;
3. the BCF that backcrosses is sowed
1seed, identifies its pollen quantity and Pollen Activity in the florescence, whole BCF
1it is sterile that plant is holandry, and the male sterile plant 4-10 that name obtains is cms4-10, its F
1male sterile plant 4-10-2,4-10-4,4-10-5,4-10-6 and 4-10-12 as cms4-10 Derivative line, called after cms4-10-2, cms4-10-4, cms4-10-5, cms4-10-6 and cms4-10-12;
4. before autumn, alfalfa was gathered in the crops, select and remain male sterile line cms4-10 as maternal plant according to parent's standard, alfalfa western giant 10-10 maternal plant is as paternal plant, according to a conventional method cuttage clonal propagation is carried out to maternal plant and paternal plant, safe overwintering, in field planting at the beginning of 3 months spring of next year in 40 ~ 60 order net isolation method districts, maternal plant and paternal plant are in the ratio field planting of 4 ~ 5:1, plantation density is line-spacing 60 cm, spacing in the rows 40 cm, honeybee is put at net indoors artificial in the florescence, maternal plant is authorized by paternal plant pollen, paternal plant is pulled out after blossom fall, seed was gathered in the brown pod phase.
Claims (2)
1. alfalfa cells matter male sterile line cms4-10 kind matter, is characterized in that: this kind of matter adopts following method to obtain:
Using bunch planting plant 3 years No. 3, the sweet agriculture of alfalfa cultivar, the sky and water, pastoral song Graze401+Z, golden queen, No. 1, public agriculture, Qingyang, the west of a river, No. 1, sweet agriculture, western giant, in Gansu Province, Jin Dalai, No. 2, sweet agriculture, giant, Algonquin, clover king, Dingxi, Long Dong and middle blue No. 1 as natural malesterile mutants screening germplasm resource bank;
(2) each alfalfa cultivars is in florescence Stochastic choice 10 strain, random selecting 5 little Hua on the different bifurcation level of every strain, play little Hua pollen on the black plastic plate of neat and tidy, adopt pollen amount fractionation testing method according to the level Four classification without pollen, a small amount of pollen, appropriate pollen, pollinia according to a conventional method, estimate pollen amount rank and record;
(3) prepare Alexander dyeing liquor:
The malachite green of I preparation 1%: the malachite of the 1g alcohol that the mass concentration of 100 ml is 95% is dissolved, and stirs 10 min, then leave standstill 3 h and get final product;
II prepares 1% acid fuchsin: dissolved by the acid fuchsin of 1 g 100 ml distilled water, to obtain final product;
III prepares 1% orange red G: dissolved by the orange red distilled water of 1 g, to obtain final product;
10 ml mass concentrations are the ethanol of 95% by V successively, the malachite green of 1 ml 1%, 50 ml distilled water, 25 ml glycerine, 5 g phenol, the acid fuchsin of 5 ml 1%, after the orange red G of 0.5 ml 1% mixes, obtain Alexander dyeing liquor;
(4) individual plant pollen can the rate of educating be measured:
Drawing 150 μ l Alexander dyeing liquors is placed on clean slide, again from 3 inflorescences of random selecting each individual plant, each inflorescence gets 2 little Hua, eject pollen respectively in Alexander dye liquor, dye 30 min, examine under a microscope pollen staining situation, if pollen can be dyed redness by dye liquor, then represent that pollen can be educated; If cyanic colours dyed by pollen envelop, then represent pollen sterility; Choose 5 visuals field, total number of fertile pollen and pollen sterile in the statistics visual field, and be calculated as follows pollen and can educate rate:
FPR=(fertile pollen sum/observe pollen frequence) × 100%;
(5) according to pollen amount classification and pollen staining rate result, if individual plant pollen amount is a large amount of or appropriate, and FPR >=10%, be expressed as male-fertile; Individual plant pollen amount be a small amount of, without pollen, or FPR<10%, be then expressed as male sterile;
To described step (1) in natural malesterile mutants screening germplasm resource bank screen, obtain the male sterile plant 4-10 of alfalfa cultivars pastoral song Graze401+Z.
2. the seed selection of alfalfa cells matter male sterile line cms4-10 kind matter as claimed in claim 1 and propagation method, comprises the following steps:
1. using the male sterile plant 4-10 of alfalfa cultivars pastoral song Graze401+Z as female parent, so that the western giant 10-10 of alfalfa can be educated for male parent, conventionally hybridize under chamber planting condition, obtain first generation of hybrid F
1seed;
2. described F is sowed
1seed, identifies its pollen quantity and Pollen Activity in the florescence, whole F
1it is sterile that plant is holandry, and using male sterile individual plant 4-10-2,4-10-4,4-10-5,4-10-6 and 4-10-12 wherein as female parent, and can educate the western giant 10-10 of parent's alfalfa and backcross, and obtains the BCF that backcrosses
1seed;
3. backcross described in sowing BCF
1seed, identifies its pollen quantity and Pollen Activity in the florescence, whole BCF
1it is sterile that plant is holandry, and the male sterile plant 4-10 that name obtains is cms4-10, its F
1male sterile plant 4-10-2,4-10-4,4-10-5,4-10-6 and 4-10-12 as cms4-10 Derivative line, called after cms4-10-2, cms4-10-4, cms4-10-5, cms4-10-6 and cms4-10-12;
4. before autumn, alfalfa was gathered in the crops, select and remain male sterile line cms4-10 as maternal plant according to parent's standard, alfalfa western giant 10-10 maternal plant is as paternal plant, according to a conventional method cuttage clonal propagation is carried out to described maternal plant and described paternal plant, safe overwintering, in field planting at the beginning of 3 months spring of next year in 40 ~ 60 order net isolation method districts, described maternal plant and described paternal plant are in the ratio field planting of 4 ~ 5:1, plantation density is line-spacing 60 cm, spacing in the rows 40 cm, honeybee is put at net indoors artificial in the florescence, maternal plant is authorized by paternal plant pollen, paternal plant is pulled out after blossom fall, seed was gathered in the brown pod phase.
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| CN113046377A (en) * | 2021-04-28 | 2021-06-29 | 吉林农业大学 | Male sterile gene MsGAL and application thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113046377A (en) * | 2021-04-28 | 2021-06-29 | 吉林农业大学 | Male sterile gene MsGAL and application thereof |
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