Separating and culturing methods for mesophyll protoplasts of petunia hybrida

Abstract

The invention discloses separating and culturing methods for mesophyll protoplasts of petunia hybrida. The separating method for the protoplasts comprises following steps: tender leaves of the petunia hybrida are weighed, processed and subjected to enzymolysis with an enzymatic hydrolysate; a CPW (cell protoplast washing) medium is added to a solution obtained after enzymolysis, the solution is filtered and centrifuged, and a supernatant is absorbed; the CPW medium is added again for washing, centrifugation is performed, precipitates are collected, and the purified protoplasts are obtained. The culturing method for the protoplasts comprises steps as follows: the prepared protoplasts are added to a protoplast culture solution and are cultured for 30 d with a solid-liquid culture method in a dark environment at the temperature of 24 DEG C, and a miniature callus formed through culture is transferred to an enrichment medium; the miniature callus is cultured for 21 d and then is transferred to a differential medium; culturing is performed. With the adoption of the separating and culturing methods, the protoplasts with high activity and high yield can be obtained, the callus can be induced through culturing of the protoplasts, shoots can be differentiated finally, an experimental material is provided for study of further somatic cell fusion and the like of the petunia hybrida, and a foundation is laid for germplasm resource innovation of the petunia hybrida.

Description

technical field [0001] The invention relates to a petunia mesophyll protoplast isolation and cultivation method. Background technique [0002] Petunia (Petunia hybrida) is a herbaceous flower of the genus Petunia in the family Solanaceae. Because of its rich flower colors and long flowering period, it is widely used all over the world and is known as the "king of flower beds". However, yellow and orange flower varieties are extremely rare, especially lack of bright and stable yellow flower varieties. At present, petunia breeding technology mostly adopts hybrid breeding. Due to the limitation of genetic resources and poor compatibility of sexual hybridization, it is difficult to breed yellow flower varieties through traditional hybrid breeding technology. Plant somatic cell fusion technology can overcome the difficulty of poor compatibility of distant hybridization, and can open up a new way for the cultivation of new varieties of petunia. [0003] The successful isolation ...

AI Technical Summary

Problems solved by technology

The reason is that the materials used are plants of the same family and different genus, with different varieties and genotypes, which lead to differences in the isolation and culture results of petunia protoplasts

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