A method for doubling of cabbage microspore haploid plants

Abstract

The invention discloses a method for doubling cabbage microspore haplobionts. The method comprises the following steps: selecting tender leaf blades growing from flower stems of microspore haplobionts, and cutting into small pieces of 1cm<2> as explants; putting the explants into an explant soaking solution, and soaking for 24h-48h; inoculating a culture medium with the explants, and inducing adventitious buds; when the adventitious buds grow to be 3.0-3.5cm high, cutting down from the roots of the adventitious buds, transferring onto a rooting culture medium for culture, thereby obtaining regeneration seedlings by induced rooting; hardening the regeneration seedlings in a room for 3-5 days, then moving into sunshade net shed in a field to be hardened for 2-3 days, then directly transplanting into a gauze shed, cultivating to promote the growth of the seedlings, in the next year, when the regeneration seedlings produce shoots and bloom, identifying ploidy to obtain doubled double haploid plants. The method not only overcomes the problems of high mortality rate and low bud induction rate easily caused by explant browning as the culture medium is added with colchicines, but also improves the induction rate of the regeneration seedlings and the doubling rate of the double haploid plants.

Description

technical field [0001] The invention belongs to the technical field of cabbage breeding, and relates to a method for doubling the microspore haploid plant to obtain a regenerated double haploid plant by using the cauline leaf of the cabbage microspore haploid plant through an in vitro tissue culture technique. Background technique [0002] Cabbage is a diploid 2n=2x=18 cross-pollination plant, and its hybrids are prepared from two parents with homozygous genotypes. Cabbage microspore culture is an important way to create parents of double haploid plants (DH strains) with homozygous genotype. In addition to many advantages, the biggest disadvantage is that haploid plants in microspore plants (Haploid plants, referred to as H plants) ratio is high, which seriously affects the haploid breeding effect of cabbage. Usually, only 10% to 50% of the regenerated plants from cabbage microspores can naturally double to form double haploid plants, which indicates that a considerable pa...

AI Technical Summary

Problems solved by technology

In this method, for the first time, the cauline leaves of flowering stems of cabbage microspore haploid flowering plants were used as explants, and double haploid plants were obtained by in vitro tissue culture after soaking in the developed colchicine and 6-BA mixed solution. It is planned to ensure that each cabbage microspore haploid plant can obtain double haploid plant regeneration, which has a significant effect compared with the method of adding colchicine to the culture medium in vitro and colchicine smearing and doubling the growth point, thus solving the problem of cabbage Haploid plants cannot be doubled or have low doubling efficiency, which greatly improves the efficiency of cabbage microspore culture

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