CN104025998A - Method for breeding corn inbred line by using hybridizing induction of haploids - Google Patents
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Abstract
The invention discloses a method for breeding a corn inbred line by using the hybridizing induction of haploids, and belongs to the field of crop breeding methods. The method comprises the following steps: pollinating basic population or cenospecies according with breeding targets by using JASS31, and selecting aleurone layer top purple and purple-free embryonic tip labeled grains; naturally or artificially doubling the obtained material, and inbreding; and selecting non-separated head progeny rows realizing consistent progenies as a new inbred line. The method has a high haploid inductivity reaching above 13%, and has the advantages of obvious selection labeling, easy screening and high accuracy; and additionally, the JASS31 used in the invention has a moderate height, a long flowering time and a large pollen amount, and natural pollination induction of haploids can be carried out, so the artificial pollination can be avoided, labor saving is realized, and it is suitable for the large scale population operation. The method also has the characteristics of high breeding efficiency, simple operation and fast breeding speed.
Description
Technical field
The invention belongs to the method for crop breeding, relate in particular to the method for utilizing hybridized induction monoploid selecting and breeding corn inbred line.
Background technology
Breeding is a periodically long job, is bred as the time that a kind generally needs 10-15, and not only workload is large, time-consuming, effort, but also may be long because lasting, the breeding objective of formulating before 10 years, after making great efforts for many years, obtain result and be often not suitable with the requirement in market.Therefore, shortening breeding time, accelerating breeding speed is the target that breeding men lay siege to always, for this reason, proposed many methods, as utilized flower pesticide or pollen to cultivate Haploid production, the method that then doubles to be bred as inbred line can shorten breeding time greatly, once be once used as on the make method, but facts have proved, because anther cultural difficulty is very large, technical sophistication, efficiency be not high, is difficult to extensive use in practice.
Nineteen fifty-nine, the Coe E H of Univ Minnesota-Twin Cities USA finds that single times of reproduction of corn induction is that Stock6 passes through hybridization and can induce maternal material production monoploid, incorporation of markings is selected, and makes to utilize hybridized induction system induction offspring to form monoploid and carries out haploid breeding and become possibility.Along with the development of corn breeding techniques, hybridized induction method for breeding haploidy is paid attention to by breeding man, become emphasis and the focus of current international breeding circle research, and some new induction systems from Stock6, are derived by the method for crossbreeding and improvement, as French SW14(Lashermes, 1988), the Krasnodar(Shatskaya of the former Soviet Union, 1994) and the ZMS(Chalyk S T of Moldova, 1994), the KWS of Germany etc.China Agricultural University utilizes Stock6 to cultivate high (Liu Zhizeng etc., 2000 of luring No. 1 of agricultural university; Chen Shaojiang etc., 2003).The value of these induction systems has substantially exceeded Stock6.
(application number is: 200610080832.4) disclose Jilin Academy of Agricultural Science and utilized M278 × Stock6 to combine as basic population, adopted pedigree method to carry out continuously selfing, the test cross seed selection of 6 generations in patent application " a kind of method of utilizing hybridized induction parthenocarpy material selective breeding corn variety ".In each generation, is chosen as master with haploid-induction, seed Navajo purple plague purpura mark (seed top aleurone layer is with purple speckle), take into account plant ABPl(plant purple simultaneously, by the control of A1A2C2BPl complementary gene, show as dominant for most corn materials) mark, pollen amount, fecundity and disease resistance successive selection.Final be bred as that average inductivity reaches 10.4%, markup good, head progeny row monosystem that Comprehensive Traits is good, name that to lure for Ji is high be No. 3.Lucky high lure be No. 3 with Navajo dominant inheritance mark, i.e. purple aleurone layer and purple plumule.With Ji high lure be No. 3 to after the pollination of other corn material, normal dliploid shows as purple aleurone layer and purple plumule point, the leaf sheath purple of plant, and the parthenocarpy seed aleurone layer purple that induction produces, plumule point is colourless, and plant leaf sheath green can be confirmed with this mark.Lucky high luring is to be for No. 3 the domestic the highest corn haploid inducing line of inductivity that select, open report, is widely used.But lucky high luring is also to have in various degree shortcoming miscellaneous No. 3: (1) plant is short and small, can not be used to carry out spontaneous pollination Haploid production in independent field.In order to overcome this shortcoming, people often will be hybridized between induction system.But may there are other some problems in the induced material that hybridization produces, such as: haploid-induction and the expression that has changed marker gene reduced.R1-nj marker gene (Nanda and Chase, 1966) (purple scultellum and purple aleurone layer apical cap) is widely used in screening monoploid from dry seeds, but, in the induced material producing in hybridization, the expression of this gene is subject to maternal influence very large, sometimes differentiate that monoploid may become very difficult, or even impossible, especially in the situation that female parent has suppressor (C1-I) to exist.(2) seed speckle mark is inconsistent to different maternal material performances, has strengthened the selection difficulty of monoploid seed.Therefore, cultivating new high-frequency induction system is the prerequisite of carrying out this work.
Summary of the invention:
The object of the invention is to provide the method for utilizing single times of selecting and breeding corn inbred line of hybridized induction.The method procedure of breeding is simple, breeding speed is fast and corn inbred line that cultivate isozygotys completely.
The present invention utilizes the method for hybridized induction monoploid selecting and breeding corn inbred line, comprises the steps:
(1), with lucky high luring be to make male parent for No. 31 to the basic population or the crossbreed pollination that meet breeding objective, gather in the crops maternal fruit ear; Therefrom screen aleurone layer top purple, plumule point is without purple, and embryo face is less, triangular in shape, the seed of the darker mark of depression;
(2), step (1) material that obtains is carried out artificial doubling or naturally doubled, the shooting stage is removed the plant that leaf sheath and blade are purple; The plant that can normally bloom after doubling is carried out to selfing, according to individual plant results self-fertility seed;
(3), plantation step (2) gained seed, select characters of progenies performance neat and consistent, do not have separative head progeny row, be new corn inbred line.
High the luring of Ji described in said method is to belong to Zea maize seed (zea mays L.) for No. 31, for the high-frequency haploid inducing line of Jilin Academy of Agricultural Science seed selection, be preserved in Chinese Typical Representative culture collection center (preservation place is: Hubei China province wuchang, wuhan Luo Jia Shan Wuhan University) on June 3rd, 2010, deposit number is: CCTCC No:P201008
Described in said method naturally doubling refer under field conditions (factors), monoploid seed or the intracellular chromosome doubling of seedling plant become normal diploid.
Artificial doubling described in said method refers to utilize colchicine solution to process monoploid seed or seedling, makes its endocellular chromosome double to become normal dliploid.
Basic population described in said method refers to by multiple inbred lines or kind after mixed pollination, the maize population of formation.This kind of groups can make multiple beneficial gene polymerizations, merit accumulation, and disease resistance is good, Comprehensive Traits is good, is therefore used as the population material of Breeding of Inbred Lines.
The material source of above-mentioned basic population can be also various good farm varieties etc.
Crossbreed described in said method refers to single cross hybrid, triple hybrid or double cross hybrid etc.
Described in said method, meet breeding objective, according to the common understanding of those skilled in the art, refer to that the aspects such as economical character, high-yield character, disease-resistant proterties and degeneration-resistant proterties, quality trait are good.
Utilize the corn inbred line that produces in said method to cultivate the method for corn hybrid seed, taking the corn inbred line cultivated in said method as one of parent, with other hybridizations between selfed lines, be bred as new crossbreed exactly.
The present invention utilizes the hybridized induction can this method Breeding of Inbred Lines of Haploid production seed, the free high-frequency hybridized induction of application inventor list doubly solid induction is---lucky high luring is the pollen of No. 31 (inductivity is up to more than 13%), give good basic population and crossbreed pollination, the seed obtaining, differentiate by chain mark property, obtain monoploid seed and plant, then through manually or naturally doubling into dliploid, the inbred line of isozygotying of selfed breeding Cheng Xin.Realize gametophyte and select, the corn inbred line that quick breeding is new, creates new germ plasm resource, generally shortens inbred line breeding time 3-4.Lucky high luring is to be for No. 31 to be that No. 3 seed selections form by improveing lucky high luring, and hybridization haploid-induction reaches 13.67%, higher than Ji high lure be No. 3 10.40%, and proper height, can be used to carry out spontaneous pollination Haploid production in independent field; The speckle mark of seed is also more obvious, lucky high luring is that the seed speckle mark of No. 3 is inconsistent to different maternal material performances, strengthen the selection difficulty of monoploid seed, and lucky high luring is the selection of having strengthened seed speckle mark for No. 31, different maternal materials is had to good seed speckle mark.
Lucky high luring is the source of No. 31:
2003, taking Ji high lure be No. 3 as maternal, the crossbreed that Stock6/M278-8-9-1-6 obtains for paternal hybrid is basic material, adopt pedigree method, be chosen as master with haploid-induction, seed Navajo purple plague purpura mark (seed top aleurone layer is with purple speckle), carry out individual plant mensuration, through continuous selfing and the test cross of 6 generations, take into account plant ABPl mark, pollen amount, fecundity and disease resistance simultaneously and select, being finally bred as lucky high the luring of high-frequency monoploid is No. 31.Lucky high luring is that No. 31 fields show as plant purple, blade purple, and flower pesticide purple, pollen amount is large, and flowering time is long.Particularly lucky high luring is to have more significantly seed Navajo genetic marker, i.e. purple aleurone layer and purple embryo labels than other induction frenulums No. 31.With Ji high lure be No. 31 to after the pollination of other corn material, normal dliploid shows as purple aleurone layer and purple plumule point, the leaf sheath purple of plant, and the parthenocarpy seed aleurone layer purple that induction produces, plumule point is colourless, and plant leaf sheath green can be confirmed with this mark.
Compared with prior art, the advantage that the present invention has and beneficial effect: the inductivity of (1), the inventive method haploid inducing line used is high, the present invention's Ji used is high to lure is that the haploid-induction of No. 31 is on average more than 13.67%, even if improving 1% at this area inductivity is all very large raising, in addition be more than 3% raising, therefore, in the present invention, lucky high luring is that the inductivity of No. 31 is far above existing induction, lure as Ji is high is that the haploid-induction that the haploid-induction of No. 3 is 10.40%, Stock6/M278-8-9-1-6 is 8.58%; (2) in the inventive method, high the luring of Ji used is to have more significantly seed Navajo genetic marker, i.e. purple aleurone layer and purple embryo labels than other induction frenulums No. 31.With Ji high lure be No. 31 to after the pollination of other corn material, normal dliploid shows as purple aleurone layer and purple plumule point, the leaf sheath purple of plant, and the parthenocarpy seed aleurone layer purple that induction produces, plumule point is colourless, plant leaf sheath green, can confirm with this mark, seed mark property is obvious, and monoploid seed is easy to discriminating, and other comprehensive agronomy proterties is good; (3), lucky high the luring of induction system used in the inventive method be No. 31 proper heights, flowering time is long and pollen amount is large, can carry out spontaneous pollination induction monoploid, removes artificial pollination from, not only saves labour, and is applicable to carrying out the operation of large-scale groups; (4), the inventive method breeding efficiency is high, luring because Ji used in the inventive method is high is that the haploid-induction of No. 31 is high, the progeny population obtaining so is under the same conditions large, the probability of selecting good inbred line increases greatly, has improved breeding efficiency; (5), the inventive method breeding cost is low, because haploid-induction is high, compared with utilizing other haploid inducing lines, needed unit breeding population is relatively little, has reduced breeding cost; (6), the purity of inbred line that is bred as of the inventive method is high, utilizes the purity of its crossbreed of preparing also high, is conducive to give full play to heterotic effect; (7), the inventive method procedure of breeding is simple, saves in conventional method selfing purge processes more than at least 4 generations of filial generation, directly by acquisition doubling monoploids, breeding time also shortens greatly.
Embodiment
Embodiment 1: utilizing lucky high luring is No. 31 hybridized induction monoploid selecting and breeding corn inbred lines
Carry out as follows:
(1), 2010 years, utilizing lucky high luring in Jilin Academy of Agricultural Science is to do the external crossbreed T628394 pollination that male parent is high to output and Comprehensive Traits is good for No. 31, fruit ear 248 fringes on results maternal plant, after threshing, obtain 42326 of seeds, according to seed mark property, being the seed that top aleurone layer and plumule point are purple, is the dliploid seed of normal hybridization; And top aleurone layer is purple, plumule point is colourless, and embryo face is less, triangular in shape, and the darker mark that caves in is " accurate monoploid " seed of the T628394 parthenocarpy that is induced out; Obtain altogether 6023 of " accurate monoploid " seeds according to above-mentioned mark, haploid-induction=accurate monoploid seed/hybridization grain number × 100%=6023/42326 × 100%=14.23% of seed.
(2), the winter in 2010, in base, Hainan, academy of agricultural sciences of Jilin Province by " accurate monoploid " the seed monoseeding filtering out, emerging, (haploid breeding is to gametophytic selection in 5847 strains, each gamete has 1 cover chromosome set, each gene exists with single form, all must the challenge of environment of accepting and the pressure of natural selection, and therefore much monoploid can not grow and deteriorates to embryoless seeds; Some monoploid seed can not germinate because of depauperation; Even if or germinate because of the too weak death of bud gesture).Shooting stage is further confirmed haplobiont according to seedling growth, plant height, length of blade, leaf sheath (sheet) color and Leaf insertion angle etc.Monoploid seedling growth is slow, plant height is low, plant is thin and weak, blade is short and compared with upper punch, leaf look shallow; Dliploid seedling growth is strong, plant is high and blade is loose, and leaf sheath (sheet) purple, confirms haplobiont 5663 strains, dliploid (hybrid strain) 184 strains.Because hybrid strain is normal dliploid, its germination vigor and growth potential are all stronger, by the precision management in meticulous program request and field, can grow up to normal plant, and therefore hybrid strain number has also just represented pseudo-monoploid kernal number.So haploid-induction=accurate monoploid kernal number-hybrid strain number/hybridization kernal number × 100%=(6023-184)/42326 × 100%=13.80%.Naturally double through field, obtain plant 455 strains of not only normally reeling off raw silk from cocoons but also normally blooming, obtain 389, selfing fruit ear by selfing, single fringe is solid be 1 to 154 not etc.Other 66 strains, because the florescence is inharmonious, do not obtain selfing fruit ear.
(3), spring in 2011, self-fertility fruit ear winter in 2010 being obtained in Hainan in Jilin Academy of Agricultural Science is planted into head progeny row, and carry out selfing, in self progeny, select neat and consistent, 24 of the new corn inbred lines that Comprehensive Traits does very well, name respectively lucky DH628394-1, lucky DH628394-2, lucky DH628394-3, lucky DH628394-4, DH628394-5, lucky DH628394-6, lucky DH628394-7, lucky DH628394-8, DH628394-9, lucky DH628394-10, lucky DH628394-11, lucky DH628394-12, DH628394-13, lucky DH628394-14, lucky DH628394-15, lucky DH628394-16, DH628394-17, lucky DH628394-18, lucky DH628394-19, lucky DH628394-20, DH628394-21, lucky DH628394-22, lucky DH628394-23, lucky DH628394-24.
The above results explanation the inventive method haploid-induction is high, and seed Navajo genetic marker is obvious, and Screening and Identification monoploid seed ratio is easier to; The procedure of breeding is short, and breeding speed is fast, breeding efficiency is high, and breeding cost is low.
Embodiment 2: utilizing lucky high luring is No. 31 hybridized induction monoploid selecting and breeding corn inbred lines
Carry out as follows:
(1), 2007, with lucky 853, lucky 854, prosperous 7-2,4 287, capital 24,773-2, yellow C, yellow early four, 4 444, wise man 461 totally 10 manually synthetic coloniesies (being called for short " yellow early four colonies ") of mixed pollination of inbred lines with yellow early four blood relationships.2010, taking " yellow early four colonies " as basic material, taking Ji high lure be No. 31 as male parent is to " early yellow four colonies " pollination, fruit ear 315 fringes on results maternal plant, set seeds altogether grain 53897, by top aleurone layer be purple, plumule point without purple, and embryo face is less, triangular in shape, the depression mark property such as darker is differentiated, obtains 7589 of " accurate monoploid " seeds.
(2), the spring in 2011, in Jilin Academy of Agricultural Science by " accurate monoploid " the seed monoseeding filtering out, 7431 strains of emerging, the shooting stage is further confirmed haplobiont 6983 strains according to seedling growth, plant height, length of blade, leaf color etc.; Plant growing way is strong, plant is tall and big, blade is loose, hybridization plant 247 strains of blade purple.Because hybrid strain is normal diploid, its germination vigor and growth potential are all stronger, by the precision management in meticulous program request and field, can grow up to normal plant, and therefore hybrid strain number has also just represented pseudo-monoploid kernal number.So the haploid-induction of this test=accurate monoploid kernal number-hybrid strain number/hybridization kernal number × 100%=(7589-247)/53897 × 100%=13.62%.Naturally double through field, obtain plant 636 strains of not only normally reeling off raw silk from cocoons but also normally blooming, obtain 463, selfing fruit ear by selfing, single fringe is solid be 1 to 202 not etc.Other 173 strains, due to the reason such as the florescence is inharmonious, do not obtain selfing fruit ear.
(3), spring in 2012, in Jilin Academy of Agricultural Science, the self-fertility fruit ear of acquisition in 2011 is planted into head progeny row, field is carefully managed and is carried out self-fertility, in self progeny, select neat and consistent, 20 of the new corn inbred lines that Comprehensive Traits does very well, name respectively lucky DH Huang 1, lucky DH Huang 2, lucky DH Huang 3, lucky DH Huang 4, lucky DH Huang 5, lucky DH Huang 6, lucky DH Huang 7, lucky DH Huang 8, lucky DH Huang 9, the yellow 10 lucky DH Huangs 11 of lucky DH, lucky DH Huang 12, lucky DH Huang 13, lucky DH Huang 14, the yellow 15 lucky DH Huangs 16 of lucky DH, lucky DH Huang 17, lucky DH Huang 18, lucky DH Huang 19, lucky DH Huang 20.
The above results explanation the inventive method haploid-induction is high, and seed Navajo genetic marker is obvious, and Screening and Identification monoploid seed ratio is easier to; The procedure of breeding is short, and breeding speed is fast, breeding efficiency is high, and breeding cost is low.
Bibliography
1、Coe?E?H.Amer?Mat,1959,93:381－382
2、Lashermes,P,M?Beckert.Theor?Appl?Genet,1988,76:405－410
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4、Chalyk?S?T.Euphytica,1994,79:13～18
5、Chase?S?S.Production?of?homozygous?diploids?of?maize?from?monoploids[J].Agron.J.,1952(44):263-267
6、Gayen?P,J?K?Maden,R?Kumar,et?al.Chromosome?doubling?in?haploids?through?colchicines[J].Maize?Genet.Coop.News?Lett.,1994(68)64-65
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Claims (6)
1. the method for utilizing hybridized induction monoploid selecting and breeding corn inbred line, is characterized in that comprising the steps:
(1), with lucky high luring be to make male parent for No. 31 to the basic population or the crossbreed pollination that meet breeding objective, gather in the crops maternal fruit ear; Therefrom screen aleurone layer top purple, plumule point is without purple, and embryo face is less, triangular in shape, the seed of the darker mark of depression;
(2), step (1) material that obtains is carried out artificial doubling or naturally doubled, the shooting stage is removed the plant that leaf sheath and blade are purple; The plant that can normally bloom after doubling is carried out to selfing, according to individual plant results self-fertility seed;
(3), plantation step (2) gained seed, select characters of progenies performance neat and consistent, do not have separative head progeny row, be new corn inbred line.
2. in accordance with the method for claim 1, it is characterized in that described naturally doubling refers under field conditions (factors), monoploid seed or the intracellular chromosome doubling of seedling plant become normal dliploid.
3. in accordance with the method for claim 1, it is characterized in that described artificial doubling refers to utilize colchicine solution to process monoploid seed or seedling, makes its endocellular chromosome double to become normal dliploid.
4. in accordance with the method for claim 1, it is characterized in that described basic population refers to by multiple inbred lines or kind after mixed pollination, the maize population of formation.
5. in accordance with the method for claim 1, it is characterized in that described crossbreed refers to single cross hybrid, triple hybrid or double cross hybrid.
6. utilize the corn inbred line producing in the method described in claim 1 to cultivate the method for corn hybrid seed, be exactly the corn inbred line cultivated in claim 1 one of be parent, with other hybridizations between selfed lines, be bred as new crossbreed.
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| CN109757365A (en) * | 2019-03-19 | 2019-05-17 | 广西农业职业技术学院 | A kind of method of quick breeding silage corn self-mating system |
| CN113455378A (en) * | 2021-08-04 | 2021-10-01 | 北大荒垦丰种业股份有限公司 | Breeding method of corn haploid induction line and application thereof |
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| CN105660381A (en) * | 2016-02-25 | 2016-06-15 | 安徽省农业科学院烟草研究所 | Method for producing large-scale induction pollen sources in maize haploid breeding |
| CN108243947A (en) * | 2018-02-13 | 2018-07-06 | 吉林省农业科学院 | A kind of breeding method using yellowish green seedling label selecting and breeding corn early stage doubled haploid |
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| CN108834885A (en) * | 2018-05-25 | 2018-11-20 | 吉林省农业科学院 | A kind of corn inbred lines method |
| CN109757365A (en) * | 2019-03-19 | 2019-05-17 | 广西农业职业技术学院 | A kind of method of quick breeding silage corn self-mating system |
| CN113455378A (en) * | 2021-08-04 | 2021-10-01 | 北大荒垦丰种业股份有限公司 | Breeding method of corn haploid induction line and application thereof |
| CN115104525A (en) * | 2022-06-28 | 2022-09-27 | 北京市农林科学院 | Breeding method combining haploid breeding with conventional breeding and induced selection of corn and application of breeding method |
| CN115316263A (en) * | 2022-08-03 | 2022-11-11 | 吉林省农业科学院 | Cultivation method of isolation material Jiju No. 1 in corn test area |
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