Method for cultivating male sterile tobacco haploids from unfertilized ovules

A technology for male sterility and ovule cultivation, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of un-pollinated ovary cultivation, yield and quality trait deterioration, and breeding problems of male sterile hybrids that have not been seen. Use limited value and other issues to achieve the effect of inhibiting the occurrence of glass seedlings, promoting clustering, and increasing the frequency of induction and plant regeneration

Inactive Publication Date: 2016-03-02
TOBACCO RES INST OF HUBEI PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the 1990s, some tobacco scientific research institutions in the United States, especially the North Carolina State University Tobacco Genetics and Breeding Project Group, did a lot of research and found that after doubling, the yield and quality traits of such haploids often deteriorate seriously, and the value of breeding utilization is limited. , and the doubling frequency of this type of haploid is very low, usually only about 1%. At present, this method is rarely used in tobacco breeding (Ren Xueliang, Li Jixin, Li Minghai. 2007. The progress of tobacco breeding in the United States. China Tobacco Journal, NO.6.57-64)
Although predecessors related to the cultivation of male sterile tobacco non-pollinated ovaries (Zhu Zhongchun, Wu Haishan. 1981. Haploid plants were cultivated from non-pollinated tobacco haploid plants. Acta Genetics, NO.1.63-65; Pan Li, Yang Tiezhao. 2000. Research on the induction of embryoid body of unpollinated ovary in tobacco. Northwest Botanical Journal, NO.1.59-63), but only in pure line varieties, no male sterile hybrids with unpollinated ovary cultured Reports, especially the research on obtaining male sterile tobacco haploid plants through unfertilized ovule culture, have not been published so far

Method used

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  • Method for cultivating male sterile tobacco haploids from unfertilized ovules
  • Method for cultivating male sterile tobacco haploids from unfertilized ovules
  • Method for cultivating male sterile tobacco haploids from unfertilized ovules

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Experimental program
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Effect test

Embodiment 1

[0038] Embodiment 1: After removing the ovary wall from the ovary of the flower buds in the five stages of K1, K2, K3, K4, K5, put into the embryoid induction medium prepared with H1 medium as the basic medium and cultivate with H2. The embryoid body induction test was carried out in the embryoid body induction medium prepared with the basic medium.

[0039] Wherein, the specific operation process of embryoid induction culture with the embryoid induction medium prepared with H1 medium as the basic medium is as follows:

[0040] (1) Disinfection treatment and inoculation of the material, put the collected flower buds in the above five periods into 0.1% mercuric solution for 7-9min disinfection, then rinse with sterile water 3-4 times, then under sterile conditions Cut off the base of the flower buds of the five stages after disinfection, squeeze out the ovary with tweezers respectively, and remove the ovary wall, and put the ovules of the five stages into five cells containing ...

Embodiment 2

[0045] Example 2: The ovaries of the flower buds in the five stages of K1, K2, K3, K4 and K5 were directly stripped of the ovules, and put into the embryoid induction medium prepared with the H1 medium as the basic medium and the H2 medium respectively. The embryoid body induction test was carried out in the embryoid body induction medium prepared for the basic medium.

Embodiment 3

[0046] Embodiment three: the ovary of above-mentioned K1, K2, K3, K4, K5 five stage flower buds is put into respectively the embryoid body induction medium prepared with H1 medium as basic medium and the H2 medium after circumcision. The embryoid body induction test was carried out in the embryoid body induction medium prepared for the basic medium. After one week of inoculation and culture of the circumcised ovary, the exposed ovules were obviously enlarged, and the ovary walls of the explants with ovary walls turned green. Thickening, no rupture of the ovary wall was observed, and there was basically no sign of enlargement of the inner ovule. At this time, the ovary wall was removed, and the ovule was still browned and died during the culture process.

[0047] In the above-mentioned three embodiments, the ovaries of the five stages of K1 to K5, when the ovary wall is removed or the ovule is directly peeled off for culture, the ovules all have swelling phenomenon, but K1 to K5...

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Abstract

The invention provides a method for culturing a male sterile tobacco haplont via an unfertilized ovule. According to the method for culturing the male sterile tobacco haplont, an ovule of which the development of an unpollinated male sterile tobacco bud is between a megasporocyte stage and a binucleate stage is used as a culture medium, after sterilization treatment, the ovule is sequentially subjected to embryoid induced culture, regeneration bud rooting culture and regenerated plant culture in a 28-DEG C illumination culture chamber, so that the male sterile tobacco haplont is finally cultured, and during the period of the culture, the illumination continuously lasts for 10 hours every day under a 1,500 Lux illumination condition. The method for culturing the male sterile tobacco haplont via the unfertilized ovule is adopted to culture male sterile tobaccos, so that the frequency of tobacco ovule embryoid induction and plant regeneration are remarkably improved.

Description

technical field [0001] The invention belongs to the technical field of tobacco breeding, and in particular relates to a method for obtaining haploids of male sterile tobacco hybrids by culturing unfertilized ovules. Background technique [0002] Haploid refers to a sporophyte that has a gametic chromosome composition. Haploid induction technology, as the main component of modern high-efficiency breeding system, has the following significant advantages: After doubling the haploid material of crops, it will become a pure line (double haploid), and after selecting the best from the best, it can quickly produce homozygous excellent hybrid combinations Excellent genetic loci, so that the breeding period is shortened by 3 to 4 years compared with conventional methods; secondly, because the genotype and phenotype of the double haploid plants are completely consistent, the frequency of misselection can be greatly reduced when screening breeding materials, and the selection rate can ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 曹景林程君奇蔡长春黎根李亚培吴成林
Owner TOBACCO RES INST OF HUBEI PROVINCE
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