Method for cultivating artificial hybridization hexaploid rape microspore regeneration plants

Abstract

The invention discloses a method for cultivating artificial hybridization hexaploid rape microspore regeneration plants. The method comprises the following steps of: taking inflorescence of the artificial hybridization hexaploid rape as a donor for microspore culture; adding buds of the hexaploid rape subjected to sterilization into a 1 / 2B5-13 culture medium, grinding into suspension, filtering and centrifuging to obtain precipitate; adding into an NLN-13 liquid culture medium and mixed solution of colchicine and active carbon sequentially to obtain microspore mixed suspension; performing heat shock treatment under the dark condition and cultivating to obtain an embryoid in the cotyledon period; inoculating into a regeneration culture medium to cultivate to directly form regeneration buds; cutting the regeneration buds and inoculating into a rooting medium to perform rooting culture; transplanting to obtain the regeneration plants; and identifying ploidy. The invention firstly discloses a method for cultivating an artificial hybridization hexaploid rape microspore. By the method, double and single ploidy groups of the hexaploid rape microspore source are cultivated for the first time; and the establishment of a linkage genetic map of ABC genome is facilitated.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for artificially hybridizing hexaploid rape microspore regeneration plants. Background technique [0002] Brassica plants have relatively important economic value in the Brassicaceae. The genus Brassica contains the six most basic variety types, including Brassica napus (B.rapa, 2n=20, AA), black mustard (B.nigra, 2n=16, BB) and cabbage (B.oleracea, 2n=18, CC) three diploid species, and Brassica napus (B.napus, 2n=38, AACC), Brassica napus (B.juncea, 2n=36, AABB) and Ethiopian mustard (B.carinata , 2n=34, BBCC) three tetraploid compound species, which is the famous Yu's triangle (UN, 1935). These Brassica plants hold great potential for genetic improvement, with unique specific genes present in each species. For example, Brassica napus has strong moisture resistance and cold resistance, especially its short growth period, these characteristics make it of ...

AI Technical Summary

Problems solved by technology

We know that the culture of free microspores of Brassica napus is relatively easy, but the culture of microspores of other crops in the genus Brassica, such as Brassica oleracea and Brassica napus, is relatively difficult, especially for varieties that are difficult to germinate

Therefore, the hexaploid rape synthesized by artificial hybridization contains the three genomes of ABC, and we cannot determine whether the simultaneous existence of the three genomes will affect the embryo emergence rate and embryo quality of microspore culture, so as to influence the quality of embryos in the future. There are obstacles in the establishment of DH (doubled haploid) populations