Chilli cytoplasm male sterile line protoplast separation purification and callus forming method

Abstract

The invention provides a chilli cytoplasm male sterile line protoplast separation purification and callus forming method, which relates to the protoplast source, separation, purification and protoplast culture and callus forming and belongs to the field of biotechnology science. The method comprises the following steps that: 1, chilli cytoplasm male sterile line aseptic seedling main leaves are used as materials for separating the protoplast; 2, young tender blades are cut and are placed into a CPW9 enzymolysis solution containing cellulase, pectinase and macerozyme, and the solution is placed onto a 27 DEG C shaking bed for dark treatment; 3, the enzymolysis liquid is filtered by a 300-mesh nylon sieve, and the protoplast is collected after centrifugation washing; and 4, the obtained protoplast is cultured into a liquid culture medium containing hormone in different concentrations, and fresh culture media are regularly added in the period until the callus is formed. In the method, the chilli male sterile materials are used for carrying out protoplast separation and culture, and important significance is realized on further studying the protoplast fusion and using the fusion technology for realizing the development of the novel chilli cytoplasm male sterile line materials.

Description

Technical field [0001] The invention is a method of biotechnology science. It is a method formed by the use of pepper cytoplasm male infertility materials for primary and purification and healing tissue. Background technique [0002] The native body refers to removing naked cells from the cell wall with mechanical or enzyme disintegration methods.Because there is no cell wall, the native body is of great significance in the practice of crop breeding and genetic breeding theory.In the practice of breeding, the successful cultivation of native quality has laid the foundation for the development of body cell hybridization. It can overcome the reproductive obstacles faced by traditional breeding methods through different types of primary quality body, create new species materials, and achieve nuclear nuclear nuclear core materials.Genome reorganization.In addition, because the native body can produce a sexual variation of body cells during the cultivation process, it can be selected ...

AI Technical Summary

Problems solved by technology

However, the main problem in the current production of heterosis breeding using CMS is the lack of good CMS lines

Since CMS traits are inherited maternally, it often takes 6-10 years to transfer the traits through sexual crossing and backcrossing, which undoubtedly limits the breeding process of CMS lines

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