Roegneria kamoji mature embryo callus induction and regeneration culture method

Abstract

The invention discloses a roegneria kamoji mature embryo callus induction and regeneration culture method. The method includes steps: (1) explant disinfection and sterilization; (2) callus induction culture; (3) callus differentiation culture; (4) induced differentiated bud rooting culture; (5) transplanting culture. The method has advantages that each seed harvested in maturity of roegneria kamoji contains a mature embryo and can be stored for years, callus induction can be realized at any time in existence of the seeds, and material acquisition time limitation is released; in callus induction with mature embryos, stripping out the mature embryos is avoided, the seeds subjected to disinfection and sterilization can be used directly, and accordingly operating steps are simplified, and difficulty in operation is lowered; by adding of 2,4-D and KT in a certain ratio, the callus induction rate can be increased to 78%; in a process from differentiation culture to regeneration culture, by adding of 6-BA and KT in a certain concentration ratio, the callus differentiation rate reaches 90% or above, and the differentiated seedling rooting rate reaches 97% or above.

Description

Technical field: [0001] The invention relates to the field of biotechnology, in particular to a culture method for inducing and regenerating mature embryo callus of A. Background technique: [0002] Goose view grass is a species under the genus Goose view grass of Gramineae. It is a common herb that can be used as feed for livestock. It has a large grass yield and high edibility, and it has the high yield, Good genes such as resistance to diseases and insect pests and stress resistance can be used as valuable resources for breeding new varieties of wheat crops. [0003] As an excellent grass species with important application value, the construction of an efficient regeneration system is crucial to the establishment of a molecular genetic system, and it is also the technical basis for the study of gene function and transgenic research using gene editing technology. At present, there is only one case of research reports on the regeneration of Goosegrass, a high-frequency cul...

AI Technical Summary

Problems solved by technology

However, there are the following problems in the cultivation of callus induction and regeneration using young embryos as explants: 1. The time for taking materials is limited by the growth period, and the young embryos of Goose View Grass can only be obtained from live embryos in the late booting stage and early heading stage of Goose View Grass. It is impossible to obtain explants at any time throughout the year, and the suitable time for taking materials can only last for about 20 days, which is extremely unfavorable for the research on the construction of an efficient regeneration system of Goose View Grass; 2. The young embryos of Goose View Grass are very small, about 0.5mm×0.5mm, it needs to be peeled off from immature seeds under sterile conditions with the help of a dissecting microscope, and the immature seeds cannot be damaged, so the operator is required to have a high level of technology, the operation is difficult, and it is not conducive to large Scale expansion

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