Method for selecting and breeding Brassica napus recessive genic-male-sterile line through microspore culture technology and cloning technology

Abstract

The invention discloses a method for selecting and breeding Brassica napus recessive genic-male-sterile (GMS) line through the microspore culture technology and the cloning technology. The method comprises the following steps: a fine variety / line is hybridized with a recessive GMS line or temporary maintaining line to obtain the F1 generation; then the Brassica napus microspore culture technology is adopted to perform inducing culture to the isolated microspores of the F1 generation plant and obtain a DH strain; and then the DH strain with the sterile character is test-crossed with the temporary maintaining line, 100% of the tester strain is sterile, the corresponding sterile DH strain is the recessive GMS line; and then the cloning technology is used to rapidly obtain a large number of plants of the sterile line which are used for the complete sterile line production. By using the method of the invention, the sterile line with stable and excellent genetic background can be rapidly obtained and used in the Brassica napus recessive GMS cross-breeding.

Description

technical field [0001] The invention relates to the field of Brassica napus breeding, in particular to a method for breeding a recessive male sterile line of Brassica napus by using microspore culture and asexual cloning techniques. Background technique [0002] The isolation technology of free microspores of Brassica napus is simple, and the efficiency of obtaining embryoid bodies and regenerated seedlings through free microspore culture technology is high, and microspore culture is an ideal method for studying the differentiation conditions of pollen cells, embryogenesis and morphogenesis mechanism. method, so this technology has been generally favored by rapeseed breeders in various countries. Over the years, many scholars have conducted extensive research on the composition of the medium for microspore culture, the size of buds, the temperature change during the microspore culture process, and the increase in the frequency of regenerated seedlings, making the microspore ...

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Problems solved by technology

[0005] However, because the recessive GMS line is controlled by three pairs of genes (aabbRfRf), the segregation population is large, and the genotypes are many and complex, resulting in a large workload for testcross and testcross species identification, and great difficulty in breeding

Utilizing conventional breeding techniques, recessive genetic male sterile lines are applied in hybrid production in the form of recessive genetic sterile homozygous dimorphic lines, that is, recessive genetic male sterile lines need to be crossed by siblings of homozygous dimorphic lines Breeding and preservation ( figure 2 ), in order to make the genetic backgrounds of the sterile and fertile plants in the homozygous two-type line consistent, it must go through more than 5 generations of brother-sister crossing, which leads to a long breeding cycle, and it is necessary to produce all sterile lines in the female parent Pulling out 50% of the fertile plants in the homozygous two-type line will affect the yield and quality of the total sterile line

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