CN103329792A - Breeding, propagating and seed-producing method of purple pakchoi cytoplasmic male sterile line - Google Patents
Breeding, propagating and seed-producing method of purple pakchoi cytoplasmic male sterile line Download PDFInfo
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- CN103329792A CN103329792A CN2013102570522A CN201310257052A CN103329792A CN 103329792 A CN103329792 A CN 103329792A CN 2013102570522 A CN2013102570522 A CN 2013102570522A CN 201310257052 A CN201310257052 A CN 201310257052A CN 103329792 A CN103329792 A CN 103329792A
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Abstract
The invention discloses a breeding, propagating and seed-producing method of a purple pakchoi cytoplasmic male sterile line. The breeding, propagating and seed-producing method comprises the following steps of: 1, carrying out distant hybridization by using a brassica juncea cytoplasmic male sterile material hau CMS as a female parent and a purple pakchoi self-incompatible line Jiuyuexian as a male parent, separating an embryo of a hybrid plant, rescuing and cultivating the embryo, then transplanting in a large land to obtain an interspecific hybrid F1; 2, carrying out continuous backcross for 6 generations by selecting a sterile plant in the interspecific hybrid F1 as a female parent and the purple pakchoi Jiuyuexian as a recurrent parent to obtain the purple pakchoi cytoplasmic male sterile line. The purple pakchoi cytoplasmic male sterile line 901A obtained by breeding has the characteristics of stable sterile performance, no influence by environments, genetic background, and the like; the purple pakchoi cytoplasmic male sterile line is planted for 10 generations continuously in 5 years and is subjected to fertility identification, so that the condition that the sterility and the sterile plant rate both are 100 percent can be proven.
Description
Technical field
The invention belongs to the vegetable breeding technical field, be specifically related to a kind of seed selection, breeding and producing method for seed of Zicaitai cytoplasmic male sterile line.
Background technology
The main path of brassicaceous vegetable heterosis utilization is self incompatible line and male sterile line, and wherein cytoplasmic male sterility is desirable pollination control system, utilizes the most extensive.The brassicaceous vegetable first generation of hybrid utilizes work to carry out widely, Asian countries particularly, as China, Japan and Korea S etc., but the method that adopts self incompatible line or self compatible line is carried out the first generation of hybrid and is produced in the majorityly, the decline of parents' vitality easily takes place and occur problem such as pseudostationary easily but this method exists selfing many generations.Utilize the cytoplasmic male sterile line production of hybrid seeds, not only can obtain high-quality filial generation.Produce the Zicaitai cytoplasmic male sterile line type that goes up utilization at present and be mainly rape pol cytoplasmic male sterility (pol cms) (pol CMS) and male sterile with radish cytoplasm (ogu CMS), but there is the unsettled shortcoming of sterility in Pol CMS cytoplasmic male sterile line; There are shortcomings such as low temperature yellow and nectary degeneration in ogu CMS.Therefore excavate and formulate new cytoplasmic male sterility type, overcome the single problem of cytoplasm, widen germ plasm resource, have great importance.
Pol CMS is the Fu Ting of Hua Zhong Agriculture University discovery in 1972, extensively utilizes in rape and brassicaceous vegetable.Ke Guilan etc. are female parent with the Ma Xibaozhibuyuyuan of Pori, are male parent with Chinese cabbage 3411-7, have bred allo-plasm of Chinese cabbage male sterile line CMS3411-7.Since Ogura1968 found ogu CMS, Chinese scholars had been carried out a large amount of backcross transformations to this sterile material, at present transformation in brassicaceous vegetables such as rape, wild cabbage, Chinese cabbage, broccoli.Fang Zhiyuan 1998 introduces Ogu CMS material after 6 parts of new improvement such as CMS625, CMS629 from the U.S., and a part broccoli inbred line is carried out transformation surplus part wild cabbage inbred line and 20 surplus 30, advantages such as the not yellow of backcross progeny performance low temperature lower blade, the sterile flower that CMS625, CMS629 transformation come opens have application promise in clinical practice.
Brassica campestris var purpurea originates from middle part, China Yangtze river basin, in Hubei Province, regional cultures such as Hunan, river are with a long history, are the important autumn and winter vegetables in this area.For many years, through China vegetables genetic breeding worker's effort, utilize abundant male sterile germ plasm resource, successively select a collection of brassica campestris var purpurea hybrid new breed, as red 2000, red assorted 50, red assorted 60, red No. 1 of Hunan, Hunan, red No. 2 of Hunan, red No. 1 of China etc.The sterile source of above Zicaitai Hybrid is rape pol CMS, and excavating new Zicaitai cytoplasmic male sterile line material is the effective way of Zicaitai breeding and hybrid.
Summary of the invention
The objective of the invention is to overcome the weak point of existing Zicaitai cytoplasmic male sterile line, the selection of the stable Zicaitai cytoplasmic male sterile line of a kind of new sterility is provided, and the breeding of this Zicaitai cytoplasmic male sterile line, producing method for seed, for the vegetables Hybrid provides new germ plasm resource.
The present invention such as utilizes the applicant the sterility of disclosed mustard type rape cytoplasmic male sterility material hau CMS has been stable, thorough and not affected by environment at advantage, by with Zicaitai " aquatic foods in September " distant hybridization and back cross breeding, obtain the stable Zicaitai cytoplasmic male sterile line of sterility.Breed more Zicaitai cytoplasmic male sterile line under the isolation condition in the fall, and it and Zicaitai " July is red " are hybridized, obtain highly purified Zicaitai male sterile Hybrid.
Above-mentioned purpose is achieved by the following technical programs:
A kind of selection of Zicaitai cytoplasmic male sterile line, it may further comprise the steps:
1) utilize mustard type rape cytoplasmic male sterility material hau CMS to be female parent, be male parent with Zicaitai self incompatible line " aquatic foods in September ", carry out distant hybridization, separate the embryo of hybrid strain and place the MS solid culture medium, place then in 25 ℃ of culturing room, shine 14h every day with 1000 ~ 1500lx light, cultivate and transplant acquisition species hybrid F in land for growing field crops after 30 days
1
2) select species hybrid F
1In sterile strain be maternal, be 6 generations of recurrent parent continuous backcross with Zicaitai " aquatic foods in September ", obtain Zicaitai cytoplasmic male sterile line.
The breeding of described Zicaitai cytoplasmic male sterile line, producing method for seed, under autumn sowing isolation condition, it is maternal adopting Zicaitai cytoplasmic male sterile line, Zicaitai " aquatic foods in September " is male parent, according to male parent: maternal row is bred described Zicaitai cytoplasmic male sterile line than the ratio that is 2: 4 ~ 6;
The Zicaitai cytoplasmic male sterile line that obtains with breeding is maternal, be male parent with Zicaitai " July is red ", and under the isolation condition in the fall, according to male parent: maternal row carries out hybrid seeding than the ratio that is 2: 4 ~ 6, acquisition Zicaitai male sterile Hybrid.
The invention has the beneficial effects as follows:
1) the Zicaitai cytoplasmic male sterile line that obtains of seed selection of the present invention has the stable characteristics of sterility, and be not subjected to the influence of environment (as low temperature or high temperature) and genetic background, in continuous 5 years 10 generations, planted and carried out fertility and identify that sterile degree and the sterile plant rate of this male sterile line are 100% in Wuhan, Hubei.
2) overcome the male sterile line yellow in seedling stage that exists in the prior art, nectary is undeveloped and have defectives such as trace-pollen.
3) program simple (removing artificial pollination and chemical treatment from) of the Zicaitai cytoplasmic male sterile line hybrid seeding of employing seed selection of the present invention, Hybrid purity height (the male sterile line sterile degree of employing is for having 100%), with the obvious advantage.
Description of drawings
Fig. 1: be technology path figure of the present invention
Fig. 2: the floral organ figure of mustard type rape cytoplasmic male sterility material hau CMS and Zicaitai cytoplasmic male sterile line 901A, the Zicaitai of seed selection of the present invention keep system " aquatic foods in September " and species hybrid F
1Among the figure: A, B are mustard type rape cytoplasmic male sterility material hau CMS floral organ figure; C, be the floral organ figure of the Zicaitai cytoplasmic male sterile line 901A of seed selection of the present invention; D is species hybrid F
1The individual plant phenotype; E is that Zicaitai keeps system's " aquatic foods in September "; F saves the species hybrid F that the back obtains through embryo
1Tissue culturing seedling.
Fig. 3: anther development cytological observation and the species hybrid F of Zicaitai cytoplasmic male sterile line 901A and maintenance system " aquatic foods in September "
1(n=28) chromosome is identified.Among the figure: A is the anther development cytological observation of Zicaitai cytoplasmic male sterile line 901A; B is species hybrid F
1Chromosome identify.
Embodiment
The present invention is described in detail below in conjunction with embodiment.
The seed selection of embodiment 1 Zicaitai cytoplasmic male sterile line
A kind of selection of Zicaitai cytoplasmic male sterile line, select mustard type rape cytoplasmic male sterile line hau CMS to do maternal, Zicaitai self incompatible line---Zicaitai " aquatic foods in September " is made male parent, save and continuous backcross by distant hybridization, embryo, obtain the Zicaitai cytoplasmic male sterile line 901A of inheritance stability.
Concrete seed selection step is as follows:
1) utilizing the disclosed novel mustard type rape male sterile material hau CMS(of inventor has been that 200510086942.7 Chinese invention patent is open at application number, this seed material on November 12nd, 2005 in China's typical culture collection center preservation, preserving number is: CCTCC NO:P200504) for maternal, with the Zicaitai self incompatible line---Zicaitai " aquatic foods in September " (seed of this breeding material is public offering before the applying date) is male parent, by distant hybridization with from embryo culture, transplant the land for growing field crops and obtain species hybrid F
1
2) 2100 species hybrid F that step 1) is obtained
1Colony carries out fertility and identifies;
3) with step 2) the species hybrid F that obtains
1The sterile strain of colony is maternal, with maintenance be that Zicaitai 9418B distant hybridization and embryo are saved, obtain Zicaitai BC1F
1
4) the Zicaitai BC1F that step 3) is obtained
1Carrying out fertility evaluation and colony and select, is that Zicaitai " aquatic foods in September " is backcrossed with maintenance again, obtains Zicaitai BC2F
1
5) the Zicaitai cytoplasmic male sterile line BC2F that obtains with step 4)
1Making female parent, is that Zicaitai " aquatic foods in September " is 4 generations of recurrent parent continuous backcross to keep, and identifies and colony's selection by fertility, obtains Zicaitai cytoplasmic male sterile line 901A.
The embryo rescue method:
1) being maternal with mustard type rape cytoplasmic male sterile line hau CMS, is that male parent is carried out distant hybridization with Zicaitai " aquatic foods in September ", obtains species hybrid F
1
2) from the species hybrid F of growth after 15-20 days
1Get angle fruit on the plant, with the ovary of protuberance in 4 ℃ of preservations;
3) at superclean bench with 70% ethanolic solution processing angle fruit 5 minutes;
4) use 0.1% mercuric chloride solution processing angle fruit 15 minutes again;
5) distilled water with sterilization washs the angle fruit three times, each about 5 minutes;
6) peel off embryo with the dissecting needle or the tweezers that disinfect;
7) embryo that disinfects is placed in the MS solid culture medium of preparation in advance, place in 25 ℃ of culturing room intensity of illumination 1000-1500lx, illumination every day 14h, namely obtain tissue culturing seedling through cultivation in about 30 days, the group that will obtain at last training seedling is transplanted to big Tanaka in good time.
The MS culture medium preparation:
In the triangular pyramidal bottle, adorn following medium: MS minimal medium (Murashige T, skoog F, 1962, A revised medium for rapid growth and bioassays with tobacco tissue culture cultures physiol plant 15:473-479), additional 6-BA1.5mg/L, NAA0.05mg/L, sucrose 30.0g/L, agar 7.0g/L adds water to 1L, transfers pH5.9-6.0.(121 ℃, high pressure steam sterilization 30 minutes) sterilization according to a conventional method, the triangular pyramidal bottle is taken out in the sterilization back from autoclave, lie in and make its cooled and solidified on the experimental bench, and is standby.
This sterile source is novel mustard type rape male sterile material hau CMS, does not report both at home and abroad.
Breeding, the production of hybrid seeds of embodiment 2 Zicaitai cytoplasmic male sterile line 901A
Under Chinese Hubei Wuhan autumn sowing (annual September) isolation condition, it is maternal adopting Zicaitai cytoplasmic male sterile line 901A, Zicaitai " aquatic foods in September " is male parent, press male parent: female parent is that the row of 2:4 is bred described Zicaitai cytoplasmic male sterile line than (the quantity ratio of row), other cultivation managements obtain more Zicaitai cytoplasmic male sterile line 901A with the Tzu Tsai tongue.
With the Zicaitai cytoplasmic male sterile line 901A after the breeding and Zicaitai " July is red " hybridization, obtain Zicaitai cytoplasmic male sterility Hybrid.
The phenotype of table 1 Zicaitai cytoplasmic male sterile line 901A and novel leaf mustard hau CMS relatively
Claims (2)
1. the selection of a Zicaitai cytoplasmic male sterile line is characterized in that may further comprise the steps:
1) utilize mustard type rape cytoplasmic male sterility material hau CMS to be female parent, be male parent with Zicaitai self incompatible line " aquatic foods in September ", carry out distant hybridization, separate the embryo of hybrid strain and place the MS solid culture medium, place then in 25 ℃ of culturing room, shone 14 hours every day with 1000 ~ 1500lx light, cultivate and transplant acquisition species hybrid F in land for growing field crops after 30 days
1
2) select species hybrid F
1In sterile strain be maternal, be 6 generations of recurrent parent continuous backcross with " aquatic foods in September ", namely obtain described Zicaitai cytoplasmic male sterile line.
2. the breeding of a Zicaitai cytoplasmic male sterile line, producing method for seed, it is characterized in that: under autumn sowing isolation condition, it is maternal adopting Zicaitai cytoplasmic male sterile line, Zicaitai " aquatic foods in September " is male parent, according to male parent: maternal row is bred described Zicaitai cytoplasmic male sterile line than the ratio that is 2: 4 ~ 6;
The Zicaitai cytoplasmic male sterile line that obtains with breeding is maternal, be male parent with Zicaitai " July is red ", and under the isolation condition in the fall, according to male parent: maternal row carries out hybrid seeding than the ratio that is 2: 4 ~ 6, acquisition Zicaitai male sterile Hybrid,
Wherein, described Zicaitai cytoplasmic male sterile line obtains according to the following steps seed selection:
1) utilize mustard type rape cytoplasmic male sterility material hau CMS to be female parent, be male parent with Zicaitai self incompatible line " aquatic foods in September ", carry out distant hybridization, separate the embryo of hybrid strain and place the MS solid culture medium, place then in 25 ℃ of culturing room, shone 14 hours every day with 1000 ~ 1500lx light, cultivate and transplant acquisition species hybrid F in land for growing field crops after 30 days
1
2) select species hybrid F
1In sterile strain be maternal, be 6 generations of recurrent parent continuous backcross with " aquatic foods in September ", namely obtain described Zicaitai cytoplasmic male sterile line.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109566407A (en) * | 2019-01-28 | 2019-04-05 | 华中农业大学 | A kind of method of compound pollination system breeding Brassica napus hybrid kind |
| CN111528100A (en) * | 2020-06-16 | 2020-08-14 | 上海市农业科学院 | Method for obtaining distant hybridization progeny of broccoli and brassica napus |
| CN115968772A (en) * | 2023-01-18 | 2023-04-18 | 东北农业大学 | Creation method of Chinese cabbage cytoplasmic male sterile line |
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| CN1868260A (en) * | 2006-06-05 | 2006-11-29 | 华中农业大学 | Method for breeding male sterile series of envelopped senvy cytoplasm |
| CN1965641A (en) * | 2005-11-18 | 2007-05-23 | 华中农业大学 | Selective breeding method for rape cytoplasmic male sterile line |
| KR20100137760A (en) * | 2009-06-23 | 2010-12-31 | 이수성 | A new interspecis and intergeneric f1 hybrid bred by using cellular male sterile line of baemoochae |
| WO2011034945A1 (en) * | 2009-09-15 | 2011-03-24 | Metabolix, Inc. | Generation of high polyhydroxybutrate producing oilseeds |
| CN102475057A (en) * | 2010-11-23 | 2012-05-30 | 华中农业大学 | Selection breeding method for a Brassica napus-turnip cabbage cytoplasmic male sterility line |
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2013
- 2013-06-25 CN CN2013102570522A patent/CN103329792A/en active Pending
Patent Citations (5)
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|---|---|---|---|---|
| CN1965641A (en) * | 2005-11-18 | 2007-05-23 | 华中农业大学 | Selective breeding method for rape cytoplasmic male sterile line |
| CN1868260A (en) * | 2006-06-05 | 2006-11-29 | 华中农业大学 | Method for breeding male sterile series of envelopped senvy cytoplasm |
| KR20100137760A (en) * | 2009-06-23 | 2010-12-31 | 이수성 | A new interspecis and intergeneric f1 hybrid bred by using cellular male sterile line of baemoochae |
| WO2011034945A1 (en) * | 2009-09-15 | 2011-03-24 | Metabolix, Inc. | Generation of high polyhydroxybutrate producing oilseeds |
| CN102475057A (en) * | 2010-11-23 | 2012-05-30 | 华中农业大学 | Selection breeding method for a Brassica napus-turnip cabbage cytoplasmic male sterility line |
Non-Patent Citations (1)
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| 万正杰等: ""新型芥菜细胞质雄性不育hauCMS在十字花科蔬菜中的利用研究"", 《中国十字花科蔬菜研究进展 2009——中国园艺学会十字花科分会第七届学术研讨会论文集》, 14 October 2009 (2009-10-14), pages 104 - 106 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109566407A (en) * | 2019-01-28 | 2019-04-05 | 华中农业大学 | A kind of method of compound pollination system breeding Brassica napus hybrid kind |
| CN111528100A (en) * | 2020-06-16 | 2020-08-14 | 上海市农业科学院 | Method for obtaining distant hybridization progeny of broccoli and brassica napus |
| CN111528100B (en) * | 2020-06-16 | 2022-07-26 | 上海市农业科学院 | Method for obtaining distant hybridization progeny of broccoli and cabbage type rape |
| CN115968772A (en) * | 2023-01-18 | 2023-04-18 | 东北农业大学 | Creation method of Chinese cabbage cytoplasmic male sterile line |
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Application publication date: 20131002 |