Culture method for directly obtaining plant by pepper anther and culture medium

Abstract

The invention relates to a culture method for directly obtaining a plant by a pepper anther and a culture medium, and belongs to the field of plant cell engineering. The culture method comprises the steps: (1), selecting a flower bud with microspores in a mid-late uninucleate stage; (2), soaking the flower bud by alcohol with the concentration being 70 percent for 0.5-1min and mercuric chloride with the concentration being1.2 percent for 8-15min, washing for 3-5 times by using sterile water, sucking water to be dry by using sterile filter paper to obtain a sterile anther in the mid-late uninucleate stage; (3), floating the sterile anther in the mid-late uninucleate stage in the liquid culture medium, carrying out dark shake culture for 20-25 days, and then carrying out illumination shake culture; (4), after carrying out illumination shake culture for 10-20 days, transferring a formed cytoledon-stage embryo to an MSO solid culture medium for being cultured, and growing into a normal plant. The culture method disclosed by the invention has the advantages that (1), the induction frequency of a normal embryo is greatly increased, the regeneration frequency of the normal plant is remarkably increased; (2), the operation process is simple, and the plant can be directly obtained from microspores freely dispersing in the liquid culture medium; (3), the culture method is suitable for multiple types of peppers such as cow-horn peppers, capsicum, line peppers and sweetbell redpeppers.

Description

technical field [0001] The invention belongs to the technical field of plant cell engineering, and in particular relates to a cultivation method and a culture medium for directly obtaining plants from capsicum anthers. Background technique [0002] Although many pepper varieties have obtained DH plants through conventional anther culture, the induction frequency is still very low overall. Therefore, if a large number of embryoid bodies can be obtained through the method of free microspore culture, and then induced to develop into normal regenerated plants, and then artificially doubled to obtain DH plants, this will greatly improve the breeding efficiency. However, to make the current free microspore culture technology a part of conventional hybrid breeding methods, there are still many problems that hinder its application in large-scale breeding; two of the most prominent problems are: [0003] (1) The frequency of microspore embryo induction in pepper is still at a low le...

AI Technical Summary

Problems solved by technology

However, to make the current free microspore culture technology a part of conventional hybrid breeding methods, there are still many problems that hinder its application in large-scale breeding; two of the most prominent problems are:

[0005] (2), capsicum microspore embryos cannot develop normally, and the ratio of normal plants in the regenerated plants is low

In the culture of capsicum microspores, the occurrence rate of malformed embryos is high, and there is a phenomenon of stagnation of globular embryos