Screening and preserving method for Lolium L. embryogenic callus

Abstract

The invention belongs to the technical field of cell engineering, and specifically relates to a method for obtaining Lolium L. embryogenic callus and maintaining regeneration capacity of the callus. According to the present invention, the material for callus induction is the mature seed embryos of the Lolium L.; the mature embryos are adopted to induce the callus, after the mature embryos are cultured for 1 month, 4 secondary cultures are performed, wherein the time of the 4 secondary cultures is 2 months, and the secondary culture is performed every 2 weeks; a differentiation regeneration culture treatment is performed on the callus, the differentiated plant is continuously subjected to the secondary culture, and the callus incapable of differentiation is discarded; the stipe meristem area of the secondary culture plant after regenerating is subjected to the second callus induction, wherein the culture time is 1 month; the callus is subjected to the secondary culture for 2 months, wherein the culture media is changed every two weeks; then the differentiation regeneration culture treatment is performed on the callus, wherein the callus incapable of differentiation is discarded; a long-term secondary culture is performed on the regenerated plant, when the callus is required to carry out the relevant research, the stipe meristem area of the plant is subjected to induction and secondary culture to obtain the required callus, wherein the regenerated plant is subjected to the second differentiation. According to the present invention, the operation is simple and feasible, and the stable and high-frequency differentiation regeneration efficiency can be obtained.

Description

technical field [0001] The invention belongs to the technical field of cell engineering tissue culture, in particular to a method for obtaining ryegrass (Lolium L.) embryogenic callus and maintaining its regeneration ability, and related ryegrass callus induction, in vitro regeneration, etc. . Background technique [0002] Among the ryegrass plants, perennial ryegrass (Lolium perenne L.) and annual ryegrass (Lolium multiflorum L.) are widely cultivated. They are planted all over the world. They are introduced and cultivated in my country. They are excellent forage grass and lawn grass. , so it is of great significance to use transgenic technology to genetically improve it. [0003] The most important foundation of transgenic technology is tissue culture technology. At present, most of the genetic transformation of ryegrass is to transform its callus, and then screen and differentiate to obtain transgenic plants. Therefore, obtaining callus with differentiation ability is t...

AI Technical Summary

Problems solved by technology

However, mature seed embryos of ryegrass are very small, with a diameter of about 0.5-1.0mm; immature embryos are even smaller, with a diameter of less than 1.0mm, and it is very difficult to obtain materials due to seasonal restrictions.

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