CN1245636A - Method for producing corn hybrid seed by utilizing distant hybridization and parthenogenetic reproduction - Google Patents
Method for producing corn hybrid seed by utilizing distant hybridization and parthenogenetic reproduction Download PDFInfo
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- CN1245636A CN1245636A CN 98117627 CN98117627A CN1245636A CN 1245636 A CN1245636 A CN 1245636A CN 98117627 CN98117627 CN 98117627 CN 98117627 A CN98117627 A CN 98117627A CN 1245636 A CN1245636 A CN 1245636A
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Abstract
The present invention provides a method for production of corn hybrid seed by using combination of distant hybridization and female parthenogenesis, aimed at solving the defects of low setting persentage and excessive vegetative growth and separation of filial generation of existent technology. The several high-quality varieties have been cultured by using said invented method.
Description
The present invention relates to produce the method for corn hybrid seed, specifically, the present invention relates to the method for utilizing distant hybridization and parthenogenesis to produce corn hybrid seed.
Subject matter on corn breeding is that the blastogenesis basis is narrow.For a long time, corn breeding is continued to use intervarietal cross and systematic breeding method always both at home and abroad, and affiliation is near, can not solve the hereditary basis poorness, and the level and the progress of breeding work are greatly limited.For the blastogenesis basis widen and create the new germ plasm resource, improve breeding level, many genetic breeding scholars concentrate on notice in the development and use of wild resource.By the distant hybridization approach gene that economic worth is arranged that wild relatives belongs to is imported cultivated maize, to create the new germ plasm of genetic diversity.
The corn distant hybridization breeding method is to make female parent with corn, belongs to sibling species and hybridizing as male parent.During hybridization paternal pollen is awarded on the filigree of female parent.The 2nd year with the first generation of hybrid (F
1) plant and corn backcross and obtain first backcross generation (BC
1).The 3rd year from BC
1In select fine individual plant and corn to backcross to obtain second backcross generation (BC
2).Until the 8th to 10 year, even backcross down so continuously for more time.In backcross process, also can adopt and backcross and selfing hockets.
Utilizing this method ripening rate very low, is example with the perennial teosinte of dliploid, and each fruit ear of ordinary circumstance is solid below 10 or lower, even not solid.Moreover the long-term mad separation of its distant hybrid offspring can not be stablized.Since the seventies, the genetic breeding scholar of China and foreign countries has just carried out the corn distant hybridization breeding research, adopt the continuous multi-generation of the classics of this routine method of backcrossing always, or give method such as backcross alternately certainly, so that the process continuous backcross is more than 8 generations, even more generations still can not obtain stablizing the offspring, so far (Petrov 1976 not obtain the good allos germplasm that the utilizes pure lines of genetic stability as yet with the method in breeding, In:S.S.khokhlov, Apomixis and Breeding, pp18-28, Amerind publishing co.pvt, ltd, New Delhi; Zhelegnova 1976, Tr.-Biol.-in-ta.-sib.-otd.-AN-SSSR.No.30.109-115; 1994 In:Biotechnology inAgricuture and Forestry.Vol.25 such as Magoja, maize (ed.by Y.P.S.Bajaj) springer-verlag BerlinHeidelberg; 1993 Sichuan Agricultural University's journals such as Niu Yingfeng, the genetics research of 1991 China such as 11 (4) 579-584 and Lai Zhongming, Chinese science publishing house, P.85-87.).For this reason, must seek new approach.
The purpose of this invention is to provide a kind of method of utilizing distant hybridization and parthenogenesis to produce corn hybrid seed.
In the prior art there be the gynecogenic method of inducing maize: (1) biological factor is induced, and it comprises between kind and the distant hybridization method, delayed pollination method, allogeneic cytoplasm method, twin seedling separating method etc.; (2) physics responsible is induced, and it comprises radiation exposure (X ray, r ray, ultraviolet ray etc.), the radiation of thermal neutron stream, Temperature Treatment etc.; (3) chemical factor is induced, and it comprises auxins, steroids, mutagen class and other class chemical substances.Its processing method has two kinds, and a kind of is that drug treating combines with pollination, and another kind is that drug alone is handled.
The parthenogenesis of past corn is induced, mainly be to pass through biological factor, as intervarietal cross and distant hybridization method, the delayed pollination method, allogeneic cytoplasm method and twin seedling back-and-forth method etc., physics responsible is as X or r radiation exposure, ultraviolet irradiation, the radiation of thermal neutron stream, Temperature Treatment etc., chemical factor, handle pollen in conjunction with pollination etc. as chemical agent, the parthenogenesis that these methods obtained is a monoploid, and its induction frequency is low, and material therefor is inbred line or intervarietal cross kind, and (Chase 1969, Bot, Rev.35 (2) 117-167; 1974, Advances and potential (Ed.K.Kasha) univ.of Guelph.Guelph, ontario, canada, P.2 11-230; Deanon 1957 philipp.Agric.41:364-377).
Use the research of chemicals induced parthenogenesis on plants such as cotton, tomato, cucumber, black-currant, report to be arranged separately.But before the eighties, the corn of one of staple food crop does not have the research of this respect as yet.
Our laboratory since the early 1980's, at first carried out chemicals inducing maize parthenogenesis Study on Technology, its purpose is inducing maize parthenogenesis monoploid, through doubling to obtain zygoid, this method is a new way efficiently to the breeding of the strong crop corn of hybrid vigour.
The method of utilizing distant hybridization and parthenogenesis to produce corn hybrid seed of the present invention may further comprise the steps:
(1) makes female parent with corn, make male parent with the dliploid teosinte, male parent was carried out the dark place reason 25-30 days before heading, cutting off filigree near the fruit ear top after filigree spues before pollination, or cut top 0.1-0.5 centimetre of fruit ear axle and filigree, after 1-3 days, when filigree grows the 0.5-1.5 centimetre again, pollinate, obtain hybrid F
1
(2) make female parent with corn, with distant hybrid F
1Backcross as male parent and to obtain first backcross generation (BC
1);
(3) at BC
1Select the good plant of corn type to carry out following processing among the plant group:
Fruit ear was weaved silk back 4-6 days, the 0.0-0.5 centimeters is cut filigree at distance fruit ear axle top, next day or will contain the 2-4% dimethyl sulfoxide (DMSO) same day when filigree grows to the 1.0-2.0 centimetre left and right sides and the 30-50ppm Malaysia acyl callosity MS aqueous solution is sprayed on the filigree, obtain parthenogenesis zygoid seed (Pa
1);
(3) from a parthenogenesis generation (Pa
1) in the plant colony to the resistance of individual plant, the comprehensive agronomy proterties is identified, chooses some fine individual plants, the solid seed of its plant is Pa
2
(4) from parthenogenesis two generations (Pa
2) in the strain system to the genetic stability of strain system, the comprehensive agronomy proterties, good pure lines are chosen in evaluations such as resistance, and prepare the test cross combination simultaneously, obtain corn hybrid seed.
Dliploid teosinte in the said method can be the perennial teosinte of dliploid.
In the said method, the time of the clipping and carving silk of the 2nd step was good with at 8 o'clock in the morning in the past, mainly prevented pollen contamination in the air; The spray liquor time is good after with at 7 o'clock in afternoon, and the first is too high for fear of temperature, reduces the too fast evaporation of soup, and it two prevents pollen contamination in the air.
In the said method, consisting of of the 2nd step adds 500-700ppm PEG (6000) in 2-4% dimethyl sulfoxide (DMSO)+30-40ppm Malaysia acyl callosity MS aqueous solution, and the solid effect of its induced parthenogenesis improves; Add 500-700ppm PEG (6000)+2-4%Tween20 in this soup, the solid effect of its induced parthenogenesis is better.
In the said method, if the medicine liquid spray of the 2nd step is twice better effects if of continuous two days internal sprayings.
The advantage of chemicals induced parthenogenesis is separately, it does not need pollina's the pollen or the stimulation of andro gamete to participate in fertilization, only depend on drug-induced generation unisexual reproduction, this has just removed biological factor recited above from, with and after the physical factor of pollination combination and several different methods such as drug-induced handle, must from hybrid, select monoploid or zygoid is differentiated, select supervisor, simplify the formality that parthenogenesis induces greatly and saved a large amount of workloads.
Handle corn inbred line, improvement kind, farmers''s kind, the crossbreed of not pollination with chemicals inducing maize parthenogenesis method of the present invention, all obtain the ratio better effect of lonely female method in the past, its induced parthenogenesis ripening rate calculates average out to 0.193% by handling Xiao Hua, be up to 0.456%, natural parthenogenesis ripening rate is 0.0005%.
The additive method parthenogenesis of inducing is a monoploid as mentioned above, and the outstanding advantage of our drug-induced corn parthenogenesis technology is, this method induced parthenogenesis directly produces zygoid, rather than monoploid, and this has just gone beyond the obstacle of corn doubling monoploids difficulty.
We are applied to improved chemicals induced parthenogenesis generation zygoid technology to have obtained good effect on the distant hybridization breeding of corn and the perennial teosinte of dliploid since the eighties.About the affiliation of perennial teosinte of dliploid and corn, there is the scholar to think two kinds of same genus, other scholars think two different genus, still do not have final conclusion so far.The drug-induced parthenogenesis technology of applied chemistry in the corn distant hybridization breeding does not still have the research of this respect so far both at home and abroad.Breeding practice by for many years proves that chemicals induced parthenogenesis technology is used for the corn distant hybridization breeding, improves and finishes real frequency, has overcome the long-term mad unsettled hard nut to crack of separating of hybrid generation, makes it at generation (Pa early
2) genetic stability gets off.We successively select the different ecological type corn work female parent of different genetic backgrounds and the distant hybrid BC that the perennial teosinte of dliploid is made paternal hybrid for use so far
1Make the donor parents of induced parthenogenesis, all can be in its parthenogenesis early for (Pa
2) obtaining the pure lines of genetic stability, this shows that this method has highly repeatability.We have selected the allos germplasm pure lines of a collection of excellence now, as, 540,540 is excellent, 445A, 515A, pt101, pt103, in 1575, in 1580 etc.They have made elite hybrid, and what have promotes on producing by authorization as losing single No. 6 kinds (5003 * 540), have as 540 * H
21, 1302 * 540,1302 * 445A, crossbreed such as 8112 * 515A are being participated in the examination of provincial district, and performance is good.Also have a collection of elite hybrid just carrying out compatibility test, plant experimentally, demonstrate in various places.Remove this, also create the new germ plasm resource of large quantities of economically valuables.Lonely female pure lines 540 grades of corn distant hybridization breeding method seed selection and the distant hybridization kind of preparation thereof are lost single No. 6 etc. and to be belonged to our initiative.
EXAMPLE Example 1
Total Test carries out at heredity institute of BeiJing ZhongKe institute test site.
We were female parent in 1985 with corn inbred line 330, were that male parent is hybridized with the perennial teosinte of dliploid.Before heading, carry out shading dark place reason in early July to early August, open from shading at 4 o'clock in afternoon at 9 o'clock in morning next day.Maternal male parent was sowed the greenhouse at the beginning of 4 months in the sowing of two phases of 6-15 bu June, transplanted the land for growing field crops before and after May 20.When maternal filigree extraction of late August, cut filigree with the neat fruit ear axle of scissors top.Treat that filigree grows pollination about 1.0 centimeters again, 5 on hybridization fruit ear obtains hybrid seed (F
1) 143, solid 28.6 of average every fringe.
Sowing on April 25th, 1 F
1Seed.With F
1Plant is a male parent, is female parent with inbred line 330, and in 8 months, phase of reeling off raw silk from cocoons of blooming in the last ten-days period hybridizes, 6 on the fruit ear backcrossed obtains first backcross generation (BC
1) 573 in seed, solid 95.5 of average every fringe;
25-26 day in April, the 3rd is sowed BC
1Seed.From BC
1Select in the plant colony that corn type growth potential is strong, healthy and strong, good plant 386 strains carry out parthenogenesis and handle, and directly produce zygoid seed (Pa
1), processing method is as follows:
After fruit ear reeled off raw silk from cocoons 4 days, cut filigree in neat fruit ear axle top, 7 o'clock Ban in the morning, when next day, filigree grew 1.0 centimeters left and right sides again, in afternoon 7 o'clock Ban with medicine liquid spray on filigree;
Handling 1 soup consists of: the 2% dimethyl sulfoxide (DMSO)+40ppm Malaysia acyl callosity MS aqueous solution, handle 242 on fruit ear, and solid 34,0.141 of every fringe average out to;
Handling 2 soups consists of: handle adding 600ppm PEG (6000) in 1 soup, handle 144 on fruit ear, and solid 43,0.299 of every fringe average out to;
The 2nd handles than 1.1 times of the 1st processing raising ripening rates, obtains 77 (Pa of parthenogenesis seed
1).
1988 with parthenogenesis Pa
1Seed was sowed April 26.From 74 strain Pa
1Identify in the plant colony and choose that the plant strain growth gesture is strong, strong stress resistance, disease-resistant, resistant to lodging, the plant type compactness, fruit ear is big, and the fringe position is moderate, and grain quality is good, and the individual plant productive potential are big to wait good individual plant 31 strains of comprehensive agronomy proterties, and its results seed is Pa
2
1989 with parthenogenesis Pa
2Seed was sowed by strain system April 26.From Pa
2Among the strain system group, be that the neat and consistent of plant is identified competition its purity height and genetic stability by strain, and the comprehensive agronomy proterties good (with above-mentioned standard with) strain system, from wherein choosing good pure lines 540 etc., and will be sheerly 540 preparation test crosses simultaneously and make up.
Program selects elite hybrid and loses single No. 6 (5003 * 540) in tests such as test cross evaluation, product compare routinely later on, be characterized in the Chinese People's Anti-Japanese Military and Political College, helminthosporium maydis, resistant to lodging, drought resisting is waterlogging, not early ageing, fruit ear is slightly grown, quality better, suitable grain is raised dual-purpose, by authorizing in production application No. 7 (H of section's examination
21* 540) wait to authorize by district's examination.Embodiment 2
We were female parent in 1987 with corn inbred line trip 9, with dliploid for many years teosinte be that male parent is hybridized.Maternal male parent was sowed the greenhouse at the beginning of 4 months in the sowing of two phases of 6-15 bu June, transplanted the land for growing field crops before and after May 20.The dark place reason is with embodiment 1.When maternal filigree extraction of late August, 0.3 centimeter at fruit ear axle top and filigree are cut with scissors.Treat that filigree grows pollination about 1.0 centimeters again, 4 on hybridization fruit ear obtains hybrid seed (F
1) 156, solid 39 of average every fringe.
Sowing on April 27th, 1988 F
1Seed.With F
1Plant is a male parent, is female parent with inbred line trip 9, hybridizes in phase of reeling off raw silk from cocoons of blooming mid or late August, and 5 on the fruit ear backcrossed obtains first backcross generation (BC
1) 634 in seed, solid 126.8 of average every fringe.
1989 with BC
1Seed was sowed in 26-28 day in April.From BC
1Select in the plant colony that corn type growth potential is strong, healthy and strong, good plant 415 strains carry out parthenogenesis and handle, and directly produce zygoid seed (Pa
1).Processing method is as follows:
After fruit ear reeled off raw silk from cocoons 5 days, in the morning 7 o'clock neat fruit ears top cut filigree.When next day, filigree grew 1.0 centimeters left and right sides again, in afternoon 7 o'clock Ban with medicine liquid spray on filigree.
The processing soup consists of: 2% dimethyl sulfoxide (DMSO)+40ppm Malaysia acyl callosity+600ppmPEG (6000) MS aqueous solution, handle 356 on fruit ear, and solid 110,0.309 of every fringe average out to.
Nineteen ninety is with Pa
1Seed was sowed April 26.From 103 strain Pa
1Identify in the plant colony and choose fine individual plant 46 strains that its results seed is Pa
2It identifies the competition content, and standard is identical with example 1.
1991 with Pa
2Seed was sowed by strain system April 28.From Pa
2Among the strain system group, press example 1 identical content, standard is identified the competition elite plant strain, from wherein choosing good pure lines, and called after 445A, 515A, and will be sheerly 445A simultaneously, the 515A pure lines are prepared test cross and are made up.
Program is identified through test cross routinely later on, selects elite hybrid section in product ratio the test of etc.ing and tries 12 (1302 * 445A) try 18 (C with section
9* 515A) participating in provincial district to try.Embodiment 3
We single 120 are female parent in nineteen ninety in the corn single cross hybrid, are that male parent is hybridized with the perennial teosinte of dliploid.Maternal male parent was sowed the greenhouse at the beginning of 4 months in the sowing of two phases of 6-15 bu June, transplanted the land for growing field crops before and after May 20.The dark place reason is with embodiment 1.When maternal filigree extraction of late August, cut filigree with the neat fruit ear axle of scissors top.Treat that filigree grows pollination about 1.0 centimeters again.5 on hybridization fruit ear obtains hybrid seed (F
1) 215, solid 43 of average every fringe.
Sowing on April 28th, 1991 F
1Seed.With F
1Plant is a male parent, single 120 being female parent in the single cross hybrid, in 8 months, phase of reeling off raw silk from cocoons of blooming in the last ten-days period hybridizes.Backcross 5 on fruit ear obtains first backcross generation (BC
1) 814 in seed, solid 162.8 of average every fringe.
1992 with BC
1Seed was sowed in 26-29 day in April.From BC
1Select in the plant colony that corn type growth potential is strong, healthy and strong, good plant 531 strains carry out parthenogenesis and handle, and directly produce zygoid seed (Pa
1).Processing method is as follows:
After fruit ear reeled off raw silk from cocoons 4 days, in the morning 7 o'clock neat fruit ears top cut filigree, when next day, filigree grew 1.0 centimeters left and right sides again, in afternoon 7 o'clock Ban with medicine liquid spray on filigree.
The processing soup consists of: 2% dimethyl sulfoxide (DMSO)+40ppm Malaysia acyl callosity+600ppmPEG (the 6000)+2%Tween20 MS aqueous solution.Handle 436 on fruit ear, solid 167, average every fringe is 0.383.
1993 with Pa
1Seed was sowed April 28.From 153 strain Pa
1Identify in the plant colony and choose fine individual plant 61 strains that its results seed is Pa
2It identifies the competition content, and standard is identical with example 1.
1994 with Pa
2Seed was sowed by strain system April 28.From Pa
2Among the strain system group, press example 1 identical content, standard is identified the competition elite plant strain, therefrom chooses in the good pure lines 1580 etc., and 1580 prepares test crosses and make up in will be sheerly simultaneously.
Later on program is being carried out tests such as test cross, product compare and is being carried out the competition of elite hybrid routinely.
Claims (7)
1, a kind of method of utilizing distant hybridization and parthenogenesis to produce corn hybrid seed may further comprise the steps:
(1) makes female parent with corn, make male parent with the dliploid teosinte, male parent was carried out the dark place reason 25-30 days before heading, cutting off filigree near the fruit ear top after filigree spues before pollination, or cut top 0.1-0.5 centimetre of fruit ear axle and filigree, after 1-3 days, when filigree grows the 0.5-1.5 centimetre again, pollinate, obtain hybrid F
1
(2) make female parent with corn, with distant hybrid F
1Backcross as male parent and to obtain first backcross generation (BC
1);
(3) at BC
1Select the good plant of corn type to carry out following processing among the plant group:
Fruit ear was weaved silk back 4-6 days, the 0.0-0.5 centimeters is cut filigree at distance fruit ear axle top, next day or will contain the 2-4% dimethyl sulfoxide (DMSO) same day when filigree grows to the 1.0-2.0 centimetre left and right sides and the 30-50ppm Malaysia acyl callosity MS aqueous solution is sprayed on the filigree, obtain parthenogenesis zygoid seed (Pa
1);
(3) from a parthenogenesis generation (Pa
1) in the plant colony to the resistance of individual plant, the comprehensive agronomy proterties is identified, chooses some fine individual plants, the solid seed of its plant is Pa
2
(4) from parthenogenesis two generations (Pa
2) in the strain system to the genetic stability of strain system, the comprehensive agronomy proterties, good pure lines are chosen in evaluations such as resistance, and prepare the test cross combination simultaneously, obtain corn hybrid seed.
2, in accordance with the method for claim 1, it is characterized in that described dliploid teosinte is the perennial teosinte of dliploid.
3, in accordance with the method for claim 1, it is characterized in that the clipping and carving silk of this method the 2nd step carried out in the past at 8 o'clock in the morning.
4, in accordance with the method for claim 1, it is characterized in that the spray liquor of this method the 2nd step carries out after 7 o'clock in the afternoon.
5, in accordance with the method for claim 1, it is characterized in that, added 500-700ppm PEG (6000) in the MS aqueous solution that contains 2-4% dimethyl sulfoxide (DMSO) and 30-40ppm Malaysia acyl callosity of this method the 2nd step.
6, in accordance with the method for claim 5, it is characterized in that, also added 2-4%Tween20 in this soup.
7, in accordance with the method for claim 1, it is characterized in that the medicine liquid spray of this method the 2nd step is twice of continuous two days internal spraying.
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Cited By (5)
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CN101843215A (en) * | 2010-06-01 | 2010-09-29 | 四川农业大学 | Hybrid seed production method for forage maize |
CN102577926A (en) * | 2012-02-20 | 2012-07-18 | 唐祈林 | Method for breeding novel waterlogging-tolerant maize variety by using special waterlogging tolerance of teosinte |
CN103081804A (en) * | 2012-12-17 | 2013-05-08 | 沈阳农业大学 | Breeding method for introducing corn parthenogernesis by using amiprofos-methyl |
CN104886147A (en) * | 2015-06-04 | 2015-09-09 | 杨宪民 | Broad-spectrum corn parthenogernesis inducer |
CN112625098A (en) * | 2020-09-04 | 2021-04-09 | 西北农林科技大学 | Gene VvMADS39 for regulating and controlling grape seed development and application thereof |
Family Cites Families (2)
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EP0300101B1 (en) * | 1987-07-20 | 1991-12-11 | Kajima Corporation | Deodorizing plant |
CN1085040C (en) * | 1997-12-12 | 2002-05-22 | 中国科学院遗传研究所 | Distant hybridization and breeding method for cotton |
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Cited By (7)
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CN101843215A (en) * | 2010-06-01 | 2010-09-29 | 四川农业大学 | Hybrid seed production method for forage maize |
CN101843215B (en) * | 2010-06-01 | 2012-07-25 | 四川农业大学 | Hybrid seed production method for forage maize |
CN102577926A (en) * | 2012-02-20 | 2012-07-18 | 唐祈林 | Method for breeding novel waterlogging-tolerant maize variety by using special waterlogging tolerance of teosinte |
CN103081804A (en) * | 2012-12-17 | 2013-05-08 | 沈阳农业大学 | Breeding method for introducing corn parthenogernesis by using amiprofos-methyl |
CN104886147A (en) * | 2015-06-04 | 2015-09-09 | 杨宪民 | Broad-spectrum corn parthenogernesis inducer |
CN104886147B (en) * | 2015-06-04 | 2017-11-14 | 杨宪民 | Broad spectrum activity corn parthenogenesis-inducing regulator |
CN112625098A (en) * | 2020-09-04 | 2021-04-09 | 西北农林科技大学 | Gene VvMADS39 for regulating and controlling grape seed development and application thereof |
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