Breeding method of cabbage type rape woad oil cytoplasm male sterile line

A technique for male sterile lines of Brassica napus and Isatis oil, which is applied to the breeding of cytoplasmic male sterile lines of Brassica napus and Isatis oil, and in the field of cytoplasmic male sterile lines. According to the report of the successful breeding of CMS line, the results of hybridization and open pollination are good, the performance is stable, and it is conducive to the propagation of sterile line seeds and the production of hybrids.

Inactive Publication Date: 2016-01-13
HUAZHONG AGRI UNIV
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] So far, there has been no report on the successful bre...

Method used

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  • Breeding method of cabbage type rape woad oil cytoplasm male sterile line
  • Breeding method of cabbage type rape woad oil cytoplasm male sterile line
  • Breeding method of cabbage type rape woad oil cytoplasm male sterile line

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Embodiment 1

[0030] Embodiment 1 Utilizes protoplast fusion to obtain Brassica napus and Isatis indigo somatic hybrids

[0031] The Brassica napus Huashuang 3 and Isatis indigo seeds used in the present invention are provided by the Rapeseed Research Office of Huazhong Agricultural University. Huashuang 3 is a double-low rapeseed variety bred by Huazhong Agricultural University (Wu Jiangsheng et al., Breeding and Research of Brassica double-low rapeseed variety Huashuang 3. Journal of Huazhong Agricultural University, 1999, 18(1): 1- 4.). Somatic hybrids of Brassica napus and Isatis indigo and their hybrid identification methods refer to Du Xuezhu (Du Xuezhu. Genetic research on hybrids between Brassica napus and Lesquerella fendleri and Isatis indigo. [Doctoral dissertation]. Wuhan: Huazhong Agricultural University Library, 2008, 具体参见文献:http: / / www.cnki.net / KCMS / detail / detail.aspx?dbcode=CDFD&dbname=cdfd0911&filename=2008203617.nh&urlid=&yx=&uid=WEEvREcwSlJHSldTTGJhYlQ1S2twUmkxOFhsbm1ncTJZ...

Embodiment 2

[0044] Example 2 Breeding of Brassica oleracea Cytoplasmic Male Sterile Line

[0045] See the technical route for the selection and breeding of the cytoplasmic male sterile line of Brassica oleracea figure 1 shown. The specific method is as follows: the identified true hybrid (the identification method is a conventional method) is crossed with Brassica napus Huashuang No. 3 to obtain the first generation of backcross (BC1), and its 4 long stamens are fused into one, and the 2 short stamens have no anther( figure 2 ), with fine powder, poor pollen dyeability ( image 3 ). The tetradynamous stamens of this sterile line transformed into a carpelized structure, the top of which was similar to the stigma, and the lower part was attached to the ovule, and the two short stamens only had filaments and did not form anthers ( Figure 4 ). The new CMS line is completely sterile (without anthers), and both hybridization and free pollination are good, and they are stable and sterile wh...

Embodiment 3

[0048] Example 3 Observation on Stamen Development of Cytoplasmic Male Sterile Line of Brassica oleracea Isatis

[0049]Get the flower buds of the cabbage-type rape Isatis oil cytoplasmic male sterile line and the maintainer line Huashuang No. 3 to make paraffin sections, the method of paraffin sections is referenced (Li Heping, 2009), edited by Li Heping, "Plant Microtechnology", Science Press, 2009, Beijing; Chapter 2 Paraffin Wax Production Method. Microscopic observation of the tissue revealed that the stamens of the CMS line of Brassica napus Isatis oil could not develop normally, and the stamen primordia differentiated and began to homologously transform into carpels ( Figure 5 ). Table 1 shows the comparison of the abortion stages of the Brassica oleracea Cytoplasmic Male Sterile Line and polCMS and oguCMS obtained in the present invention.

[0050] Table 1 Different CMS abortion periods

[0051]

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Abstract

The invention belongs to the technical field of rape molecular breeding, and specifically relates to a breeding method of cabbage type rape woad oil cytoplasm male sterile line. The invention also relates to a protoplast fusion technology to recombinate the cytoplasm and nuclear genome of cabbage type rape-woad so as to establish cytoplasmic male sterility of recombinant cytoplasm. The breeding process comprises the following steps: obtaining the leaf meat protoplast of cabbage type rape-woad through separation and extraction, obtaining the protoplast fused somatic cell hybrid of cabbage type rape-woad through a cell fusion method; taking the obtained somatic cell hybrid of cabbage type rape-woad and leaf meat protoplast cell hybrid of cabbage type rape-woad as the female parent, taking cabbage type rape as the male parent, carrying out multi-generation backcross, and obtaining cytoplasmic male sterile line with carpelloid stamen; wherein the mitochondrion gene PCR amplification verifies that mitochondrial genome carries out recombination during the fusion process, thus the nucleoplasm becomes uneven, and the sterile character appears.

Description

technical field [0001] The invention belongs to the technical field of rapeseed molecular breeding, and in particular relates to a method for selecting a cytoplasmic male sterile line of cabbage-type rapeseed. The invention also relates to protoplast fusion technology to recombine the cytoplasmic and nuclear genomes of Brassica napus-Isatis indigo to create a cytoplasmic male sterile line with recombined cytoplasm. The invention is related to the molecular breeding technology of rape-woad hybridization. Background technique [0002] Brassica napus is an important oil crop, showing obvious heterosis. The use of cytoplasmic male sterile line (CMS for short), maintainer line and restorer line to produce hybrids is the main way to utilize heterosis in rapeseed at home and abroad. At present, polCMS and oguCMS are widely used in rapeseed hybrid breeding at home and abroad. Among them, oguCMS is widely used abroad, while polCMS is unstable in sterility, and it is easy to produce...

Claims

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Application Information

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IPC IPC(8): A01H4/00A01H1/02A01H1/04
Inventor 李再云康雷杜雪竹
Owner HUAZHONG AGRI UNIV
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