CN102224801A - Rapid multi-target property polymerization breeding method for rape - Google Patents
Rapid multi-target property polymerization breeding method for rape Download PDFInfo
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Abstract
The invention provides a rapid multi-target property polymerization breeding method for rape, belonging to a plant breeding method, and comprising the following steps of: selecting excellent breeding target properties requiring to be polymerized, preparing single-cross combinations at room temperature in winter locally, preparing multi-parent complicated cross combinations in summer at different places, and assembling and polymerizing the multiple parents with excellent target properties based on the breeding requirements; performing microspore seedling cultivation on the complex filial generation for which multi-target property breeding restructuring polymerization is already realized, constructing a dihaploid (DH) breeding group, and realizing rapid fixation and homozygosis of the target genotypes of the generation of the multi-target property polymerization; and performing effective field and indoor identification and screening on the target genotypes of the generation for which the multi-target property polymerization is realized based on the property characteristics, performing molecular marker assistant selection on important target properties, and breeding new materials conforming to the breeding targets.
Description
Technical field
The present invention relates to the rapid polymerization breeding of rape multiple target character, belong to plant breeding method, be specifically related to the fixing fast of the strange land assembling of multiple target character and polymerization, genes of interest type and isozygoty and selections such as effective evaluation of polymerization proterties and screening.
Background technology
Rape is the important oil crop of China, and cultivated area is about 100,000,000 mu throughout the year, and gross output surpasses 1,100 ten thousand tons, produces more than 440 ten thousand tons of rapeseed oils, and more than 600 ten thousand tons of high-quality protein grouts are provided, and cultivated area and gross output all occupy first place in the world.Recent two decades comes by wideling popularize high yield, double-low rapeseed new varieties and efficiency cultivation technology for high thereof, and China's rape per unit area yield significantly promotes, and gross output significantly increases, and quality is obviously improved.China's rapeseed breeding is through several generations' effort, on the basis of autonomous innovation, constantly introduce, absorb advanced foreign technology and experience, formed the breed breeding theory and technology system of distinct Chinese characteristics, be in world lead level at aspects such as heterosis utilization, the innovation of distinguished germ plasm material and anti-stalk break researchs.
But compare with American-European advanced country, at present China's rapeseed breeding substantially still is in traditional experience breeding phase, and breeding technique is backward relatively, and per unit area yield is on the low side, and productivity effect is not high, and disease-resistant, worm, crop smothering and resistance remain further to be improved.At first, the efficient of China's rape molecular mark technology is also very low, and transgenic technology does not also enter the practicability stage, and the breeding for quality technology is further improved and improves.Secondly, China's rape per unit area yield still is in reduced levels, compares with Europe to also have big gap.The 3rd, be fit to that short bar that mechanization produces anti-ly falls, kipeming high yield, anti-ly anti-ly split the angle breeding technology is that all right ripe, breeding material is comparatively poor, and breed breeding is at the early-stage.The 4th, rape is disease-resistant, worm, grass, adverse circumstance research are backward relatively, and rape drought-resistance, anti-stain, eurytopicity research such as freeze proof are deep not enough.The 5th, the transformation of scientific and technical result is still waiting to strengthen, and high-quality two low kind and high-sulfur glucoside, high erucic acid kinds are also deposited, and lack strict regional layout, and commodity vegetable seed quality remains further to be improved.Therefore, China's rapeseed breeding latest development trend is on existing science accumulation basis at present, make full use of abundant germ plasm resource, research breeding new method is strengthened the construction of GENERALIZATION OF MODERN BREEDING TECHNIQUE system, adopt biotechnology to combine with routine techniques, technology such as innovation heterosis utilization, molecular mark and microspores culture, the pyramiding breeding platform of structure multiple target character is carried out the molecule aggregation breeding, accelerate breeding process, improve breeding efficiency.The strong advantage kind of breeding high-yield, floorboard with high oil content, the improved seeds of seed selection strong stress resistance, wide adaptability, seed selection adapts to the light efficient kind of simplifying cultivation, mechanized operation, is the fundamental way that further develops China's rape industry, improves China's rape productivity effect.And the establishment fine germplasm resources enlarges rape genetic background, sets up multiple target character rapid polymerization breeding technique efficiently, is to cultivate high-quality, high yield, wide key technology suitable, how anti-new rape variety to support.
The present invention uses multi-parent strain complex hybridization, greenhouse, locality (Jiangsu) and adds generation, strange land (Qinghai) and expand multiple technologies and means such as numerous, microspores culture and molecular marking supplementary breeding summer, research and establishment the pyramiding breeding technical system of rape multiple target character fast and efficiently, provide effective ways for the new rape variety seed selection realizes the multiple target character rapid polymerization.
Summary of the invention
Technical problem
The present invention can remedy the deficiency of existing patent or technology, and a kind of rapid polymerization breeding new method of rape multiple target character is provided.
Technical scheme
1, a kind of rapid polymerization breeding method of rape multiple target character comprises step:
The strange land assembling and the polymerization of A, multiple target character: excellent breeding objective proterties that need to select polymerization, the single cross combination that winter, preparation earlier had two proterties in local greenhouse, numerous each single cross combination of strange land summer then, and be mixed with complex hybridization combination with a plurality of parents, need assemble and polymerization by breeding the parent of multiple target character;
B, genes of interest type fast fixing with isozygoty: the 2nd year autumn is with multi-parent strain complex hybridization combination F
1At local rapeseed breeding field direct-seeding planting, the spring rape initial bloom stage was got F in the 3rd year
1Plant children flower bud carries out the microspore seedling to be cultivated, and plants the engagement alms bowl after autumn monoploid microspore seedling being soaked root 30min with 2% colchicine solution then, grows to the 5-6 leaf phase after living to be transplanted to the land for growing field crops, and acquisition double haploid DH strain is a colony;
The effective evaluation and the screening of C, polymerization proterties: the 4th year spring, strain was that according to target proterties is carried out field and indoor identification, screening to DH, or objective trait is carried out molecular marker assisted selection, the seed selection polymerization breeding new material of a plurality of objective traits.
By said method, the seed selection polymerization cabbage type rape new material of short bar, high oil and CMS retention performance.
It is characterized in that:
A: then late December will be in the locality the peaceful B7 of cytoplasmic male sterility maintenance line and floorboard with high oil content strain HOC2, short bar strain APL03 by vernalization carry out adding generation to the greenhouse from the live field-transplanting of rapeseed breeding, make rape advance to the early Febuary in the 2nd year in the flowering stage in greenhouse, prepare peaceful B7 * HOC2 and peaceful two single cross hybrid combinations of B7 * APL03 after rape is bloomed respectively, gather in the crops F mid-March
1Seed; Arrived the numerous F of strange land Jia Daixia the same year by the end of March at the beginning of 4 months
1, the normal spring sowing, (peaceful B7 * HOC2) * (complex hybridization F is gathered in the crops in the multi-parent strain complex hybridization combination of peaceful B7 * APL03) by the end of September in rape preparation in flowering stage at the beginning of 8 months by the end of July
1Seed returns local normal autumn sowing;
B: the 2nd year autumn is with the multi-parent strain complex hybridization combination (peaceful B7 * HOC2) * (F of peaceful B7 * APL03)
1At local rapeseed breeding field direct-seeding planting, the spring rape initial bloom stage was got F in the 3rd year
1Plant children flower bud carries out the microspore seedling and cultivates: win the inflorescence of first flower of firm opening at 7 o'clock in morning every day, be placed on 4 ℃ of constant temperature refrigerator 24h with the moistening processing of blotting paper, the bud of selecting length next day and be 4.0mm-5.0mm is with 70% alcohol-pickled 30s, 1% mercuric chloride surface sterilization 10min, aseptic water washing 3 times, each flushing 2min, the bud of will sterilizing places sterile glass tube, adds B
5Extract 1ml smashs bud to pieces along tube wall with glass bar and to extrude microspore, and through 400 order membrane filtrations, the centrifugal 5min of 1000rpm/min adds B after removing supernatant again
5Extract suspension microspore, the multiple weight 1 time, remove supernatant, adding the NLN medium that has added 13% sucrose changes in the triangular flask, 24h are cultivated in 32 ℃ of heat shocks, and 25 ℃ of static cultivations of dark are when visible small particles embryoid occurring, go to 25 ℃ of constant temperature shaking table 50r/min shaken cultivation, treat that embryoid length forwards B to when reaching 4mm-6mm
5On the solid culture medium, 25 ℃ of illumination cultivation, every day, light application time was 16 hours, until Cheng Miao, plant the engagement alms bowl after autumn monoploid microspore seedling being soaked root 30min with 2% colchicine solution then, grow to the 5-6 leaf phase after living to be transplanted to the land for growing field crops, obtaining double haploid DH strain is colony;
C: back selected plant height is bloomed in the fertile plant bagging selfing of 140cm in spring DH strain in the 4th year system, and with the peaceful A7 of cytoplasmic male sterile line be that maternal test cross is identified extensive guarantor's relation, whole bagging selfed seeds of results and test cross F after the rape maturation
1, the male parent selfed seed is measured seed oil content with the Soxhlet method, and the autumn in the same year is with F
1Plant in pairs with corresponding test cross male parent, spring next year is identified F
1Proterties such as the plant height of fertility and male parent, oil content are selected F
1Performance male sterile, test cross male parent plant height is lower than 140cm, oil content is an individual plant greater than 45% DH strain, be breed have short bar, cabbage type rape new material that three objective traits of high oil and CMS retention performance condense together.
Beneficial effect
The present invention can assemble fast and effectively to a plurality of special proterties of rape, polymerization, fixing with isozygoty, will shorten the breeding cycle of rape variety seed selection greatly, improve breeding of new variety efficient, will play positive impetus to lifting China rapeseed breeding level.To multiple target character polymerization offspring's genes of interest type by the proterties characteristic in the field with indoorly effectively identify and screen, application molecular labelings such as important goal proterties such as cytoplasmic male sterility recovery proterties carry out assisted Selection, improve the efficiency of selection and the accuracy rate of correlated traits.
Description of drawings
Figure is the schematic diagram of rape multiple target character rapid polymerization breeding method.
Embodiment
Embodiment: the seed selection of the short bar of cabbage type rape, high oily CMS male sterile line.
A:2006 late December will be by the peaceful B7(Pu Huiming of the cytoplasmic male sterility maintenance line of vernalization, relative is deposited button, Fu Shouzhong, Zhang Jiefu, 5 beginnings are beautiful, Gao Jianqin, Chen Xinjun, the backcross breeding of the two peaceful A7 of low male sterile line of seed selection research I.MICMS of the peaceful A7 of Wild cabbage type double-low rapeseed cytoplasmic male sterile line, Jiangsu agricultural journal, 2001,17 (2): 69-72) and floorboard with high oil content strain HOC2(Fu Shou second month in a season, Zhang Jiefu, relative is deposited button, Pu Huiming, Gao Jianqin, Chen Xinjun, Chen Feng, cabbage type rape floorboard with high oil content germplasm seed selection research, Chinese oil crop journal, 2008,30 (3): 279-283), (relative is deposited button to short bar strain APL03 for Fu Shouzhong, Zhang Jiefu, Pu Huiming, Gao Jianqin, Chen Xinjun, the seed selection of cabbage type rape petalless new germ plasm APL03 of short stem, plant genetic resources journal 2006,7 (4): 468-470) carry out adding generation in Nanjing to the greenhouse from the live field-transplanting of rapeseed breeding, the greenhouse day temperature remains on the 20-25 degree, and nocturnal temperature remains on about 15 degree, and add the light processing, make the greenhouse rape advance in early Febuary, 2007 in flowering stage.After the greenhouse rape is bloomed, prepare peaceful B7 * HOC2 and peaceful two single cross hybrid combinations of B7 * APL03 respectively, gather in the crops F mid-March
1Seed.Arrived the numerous F of Qinghai Jia Daixia the same year by the end of March at the beginning of 4 months
1, the normal spring sowing, (the peaceful B7 * HOC2) * (the multi-parent strain complex hybridization combination of peaceful B7 * APL03) of rape preparation in flowering stage at the beginning of 8 months by the end of July.In this complex hybridization combination, comprise 3 important goal proterties that need polymerization: the maintenance proterties (conservation rate is 100%) of peaceful B7 maintenance line pair cell matter male sterile line, the floorboard with high oil content proterties of HOC2 (average oil content is 50.01%), the short bar proterties of APL03 (average plant height is 135.3cm).Gather in the crops complex hybridization F by the end of September
1Seed returns Nanjing and carries out normal autumn sowing.
The B:2007 autumn is with the multi-parent strain complex hybridization combination (peaceful B7 * HOC2) * (F of peaceful B7 * APL03)
1At rapeseed breeding field, Nanjing direct-seeding planting, allow plant fully grow, multiple branch.The spring rape initial bloom stage was chosen growth potential F preferably in 2008
1The plant bud carries out the microspore seedling and cultivates: win the inflorescence of first flower of firm opening in about at 7 o'clock in morning every day, be placed on 4 ℃ of constant temperature refrigerator 24h with the moistening processing of blotting paper.Select length next day and be bud about 4.5mm with 70% alcohol-pickled 30s, 1% mercuric chloride surface sterilization 10min, aseptic water washing 3 times washes 2min at every turn.The bud of will sterilizing places sterile glass tube, adds B
5Extract 1ml smashs bud to pieces along tube wall with glass bar and to extrude microspore, and through 400 order membrane filtrations, the centrifugal 5min of 1000rpm/min adds B after removing supernatant again
5Extract suspension microspore, the multiple weight 1 time removed supernatant, adds NLN-13 (13% sucrose) culture fluid and changes in the triangular flask.24h are cultivated in 32 ℃ of heat shocks, and 25 ℃ of static cultivations of dark when visible small particles embryoid occurring, go to 25 ℃ of constant temperature shaking table 50r/min shaken cultivation, treat that embryoid length forwards B to when reaching the 5mm left and right sides
5On the solid culture medium, 25 ℃ of illumination cultivation, every day, light application time was 16 hours, until Cheng Miao.Plant the engagement alms bowl after autumn monoploid microspore seedling being soaked root 30min with 2% colchicine solution then, grow to the 5-6 leaf phase after living to be transplanted to the land for growing field crops, obtaining the DH strain is colony.
The C:2009 spring rape back of blooming selects plant height in the colony to be lower than the fertile plant bagging selfing of 140cm, and with the peaceful A7(Pu Huiming of cytoplasmic male sterile line, relative is deposited button, Fu Shouzhong, Zhang Jiefu, 5 beginnings are beautiful, Gao Jianqin, Chen Xinjun, the backcross breeding of the two peaceful A7 of low male sterile line of seed selection research I.MICMS of the peaceful A7 of Wild cabbage type double-low rapeseed cytoplasmic male sterile line, Jiangsu agricultural journal, 2001,17 (2): 69-72) test cross is identified extensive guarantor's relation, whole bagging selfed seeds of results and test cross F after the rape maturation
1, the male parent selfed seed is measured seed oil content with the Soxhlet method.In more than 350 DH strain systems, the selected plant height is in plant 40 strains and peaceful A7(female parent of 140cm) test cross, the autumn in the same year is with 40 F
1With corresponding test cross male parent (DH strain system) plantation in pairs, identify F
1Proterties such as the plant height of fertility and corresponding male parent (DH strain system), oil content.Spring rape in 2010 is bloomed, and the survey showed that for the back fertility, have 17 test cross combination offsprings and show as male sterile, sterile proterties is consistent with the peaceful A7 of former male sterile line, after the rape maturation test cross offspring is shown as male sterile corresponding test cross male parent (DH strain system) and measure oil content and plant height, average oil content is 44.25%, luffing is at 39.92-48.53%, individual plant greater than 45% has 6 strains, plant height is lower than 140cm, be institute's seed selection polymerization short bar, the cabbage type rape new material of high oil and CMS retention performance: plant height is lower than 140cm, performance male sterile, oil content is greater than 45% cabbage type rape.Maintenance line S178 plant height 137.9cm wherein, oil content 47.28%.
Use this method and only use the time more than 4 years just the maintenance proterties (conservation rate is 100%) of short bar, two proterties of high oil and pair cell matter male sterile line to be carried out polymerization fast and effectively, bred the short bar that genotype is isozygotied, high oil, CMS maintenance line.Use the short bar of new seed selection, high oily maintenance line the peaceful A7 male sterile line of the low oil of former high bar is improved, seed selection obtains short bar, high oil, CMS male sterile line, lays a good foundation for cultivating the cross-bred rape new varieties that are fit to the entire mechanization operation.
Claims (2)
1. the rapid polymerization breeding method of a rape multiple target character comprises step:
The strange land assembling and the polymerization of A, multiple target character: excellent breeding objective proterties that need to select polymerization, the single cross combination that winter, preparation earlier had two proterties in local greenhouse, numerous each single cross combination of strange land summer then, and be mixed with complex hybridization combination with a plurality of parents, need assemble and polymerization by breeding the parent of multiple target character;
B, genes of interest type fast fixing with isozygoty: autumn sowing is with multi-parent strain complex hybridization combination F
1At local rapeseed breeding field direct-seeding planting, next year the spring rape initial bloom stage get F
1Plant children flower bud carries out the microspore seedling to be cultivated, and plants the engagement alms bowl after autumn monoploid microspore seedling being soaked root 30min with 2% colchicine solution, grows to the 5-6 leaf phase after living to be transplanted to the land for growing field crops, and acquisition double haploid DH strain is a colony;
The effective evaluation and the screening of C, polymerization proterties: strain is that according to target proterties is carried out field and indoor identification, screening to DH, or objective trait is carried out molecular marker assisted selection, the seed selection polymerization breeding new material of a plurality of objective traits.
2. the application of method according to claim 1 aspect the CMS maintenance line of the short bar of seed selection cabbage type rape, high oil is characterized in that:
A: then late December will be in the locality the peaceful B7 of cytoplasmic male sterility maintenance line and floorboard with high oil content strain HOC2, short bar strain APL03 by vernalization carry out adding generation to the greenhouse from the live field-transplanting of rapeseed breeding, make rape advance to the early Febuary in the 2nd year in the flowering stage in greenhouse, prepare peaceful B7 * HOC2 and peaceful two single cross hybrid combinations of B7 * APL03 after rape is bloomed respectively, gather in the crops F mid-March
1Seed; Arrived the numerous F of strange land Jia Daixia the same year by the end of March at the beginning of 4 months
1, the normal spring sowing, (peaceful B7 * HOC2) * (complex hybridization F is gathered in the crops in the multi-parent strain complex hybridization combination of peaceful B7 * APL03) by the end of September in rape preparation in flowering stage at the beginning of 8 months by the end of July
1Seed returns local normal autumn sowing;
B: autumn sowing is with the multi-parent strain complex hybridization combination (peaceful B7 * HOC2) * (F of peaceful B7 * APL03)
1At local rapeseed breeding field direct-seeding planting, the spring rape initial bloom stage was got F in the 3rd year
1Plant children flower bud carries out the microspore seedling and cultivates: win the inflorescence of first flower of firm opening at 7 o'clock in morning every day, be placed on 4 ℃ of constant temperature refrigerator 24h with the moistening processing of blotting paper, the bud of selecting length next day and be 4.0mm-5.0mm is with 70% alcohol-pickled 30s, 1% mercuric chloride surface sterilization 10min, aseptic water washing 3 times, each flushing 2min, the bud of will sterilizing places sterile glass tube, adds B
5Extract 1ml smashs bud to pieces along tube wall with glass bar and to extrude microspore, and through 400 order membrane filtrations, the centrifugal 5min of 1000rpm/min adds B after removing supernatant again
5Extract suspension microspore, the multiple weight 1 time, remove supernatant, adding the NLN medium that has added mass ratio 13% sucrose changes in the triangular flask, 24h are cultivated in 32 ℃ of heat shocks, and 25 ℃ of static cultivations of dark are when visible small particles embryoid occurring, go to 25 ℃ of constant temperature shaking table 50r/min shaken cultivation, treat that embryoid length forwards B to when reaching 4mm-6mm
5On the solid culture medium, 25 ℃ of illumination cultivation, every day, light application time was 16 hours, until Cheng Miao, plant the engagement alms bowl after autumn monoploid microspore seedling being soaked root 30min with 2% colchicine solution then, grow to the 5-6 leaf phase after living to be transplanted to the land for growing field crops, obtaining double haploid DH strain is colony;
C: spring DH strain in the 4th year is that rape is bloomed back selected plant height in the fertile plant bagging selfing of 140cm, and with the peaceful A7 of cytoplasmic male sterile line be that maternal test cross is identified extensive guarantor's relation, whole bagging selfed seeds of results and test cross F after the rape maturation
1, the male parent selfed seed is measured seed oil content with the Soxhlet method, and the autumn in the same year is with F
1Plant in pairs with corresponding test cross male parent, spring next year is identified F
1The plant height of fertility and male parent, oil content proterties are selected F
1Performance male sterile, test cross male parent plant height is lower than 140cm, oil content is an individual plant greater than 45% DH strain, be breed have short bar, cabbage type rape new material that three objective traits of high oil and CMS retention performance condense together.
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| CN102907315A (en) * | 2012-10-11 | 2013-02-06 | 湖北大学 | High-yield new-strain selective breeding method of industrial oil crop crambeabyssinica |
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| CN102948365A (en) * | 2012-11-29 | 2013-03-06 | 江苏省农业科学院 | Method for increasing hybrid rape planting purity by applying herbicide resistant mutant character |
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| CN109273052A (en) * | 2018-09-13 | 2019-01-25 | 北京百迈客生物科技有限公司 | A kind of genome monoploid assembling method and device |
| CN109273052B (en) * | 2018-09-13 | 2022-03-18 | 北京百迈客生物科技有限公司 | Genome haploid assembling method and device |
| WO2020112479A1 (en) * | 2018-11-27 | 2020-06-04 | The United States Of America, As Represented By The Secretary Of Agriculture | Production of haploid lolium |
| US11154022B2 (en) | 2018-11-27 | 2021-10-26 | The United States Of America, As Represented By The Secretary Of Agriculture | Production of haploid lolium |
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| CN111011131B (en) * | 2020-01-03 | 2021-04-20 | 河北省农林科学院遗传生理研究所（河北省农林科学院农产品质量安全研究中心） | Breeding method combining rapid development technology, molecular marker technology and conventional breeding of wheat |
| CN111011131A (en) * | 2020-01-03 | 2020-04-17 | 河北省农林科学院遗传生理研究所（河北省农林科学院农产品质量安全研究中心） | Breeding method combining rapid development technology, molecular marker technology and conventional breeding of wheat |
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