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Method for culturing male sterile tobacco haplont via unfertilized ovule

A technology for male sterility and ovule culture, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of deterioration of yield and quality traits, limited value of breeding and utilization, no public reports, etc. Clustering, increasing the frequency of induction and plant regeneration, overcoming the effect of browning

Inactive Publication Date: 2014-04-16
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 involved the cultivation of male sterile tobacco non-pollinated ovaries (Zhu Zhongchun, Wu Haishan. 1981. Haploid plants were cultivated from non-pollinated ovaries of 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 culturing male sterile tobacco haplont via unfertilized ovule
  • Method for culturing male sterile tobacco haplont via unfertilized ovule
  • Method for culturing male sterile tobacco haplont via unfertilized ovule

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

[0040] (1) Disinfection treatment and inoculation of materials, put the flower buds collected in the above five periods into 0.1% mercuric chloride solution for disinfection for 7-9 minutes, then rinse with sterile water for 3-4 times, and then in aseptic conditions Cut off the bases of the flower buds of the five stages after disinfection, extrude the ovary with forceps, and remove the ovary wall and put th...

Embodiment 2

[0045] Embodiment two: the ovary of flower bud of five stages of K1, K2, K3, K4, K5 is directly peeled off ovule, and puts into respectively the embryoid body induction medium prepared with H1 medium as basic medium and with H2 medium The embryoid body induction test was carried out in the embryoid body induction medium prepared as the basic medium. The specific operation process was as in Example 1, and the test results obtained were the same as in Example 1.

Embodiment 3

[0046] Embodiment 3: The ovaries of flower buds in the above five stages of K1, K2, K3, K4, and K5 are circumcised and put into embryoid body induction medium prepared with H1 medium as basic medium and H2 medium respectively. 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, the exposed ovules could be observed to expand obviously, and the ovary wall of the explant with ovary wall turned green Thickened, no ovary wall rupture was observed, and the internal ovules basically showed no sign of expansion. At this time, the ovary wall was removed, and the ovules were still browned and died during the culture process.

[0047] In the above three examples, the ovules in the five stages from K1 to K5 were cultured after removing the ovary wall or directly peeling off the ovules, and the ovules all had swelling phenomenon, but it can be seen from the first and second implement...

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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 Applications(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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