Mulberry transgenic method based on agrobacterium tumefacien mediated hybernacle injection

Abstract

The invention discloses a mulberry transgenic method based on agrobacterium tumefacien mediated hybernacle injection. According to the method, a trace amount of agrobacterium tumefacien engineering bacterium conversion liquid containing target gene recombinant vectors is directly injected into mulberry hybernacles to sprout; the hybernacles infect hybernacle top end meristematic tissue meristematic cells and hybernacle axillary bud meristematic cells to obtain successfully converted bud meristematic cells; the bud meristematic cells differentiate and grow to obtain transgenic transgenic branches and buds; then, transgenic grafting transgenic seedlings or transgenic mulberry seeds are obtained through the mulberry grafting vegetative propagation or female mulberry fruit and male flower hybridization sexual propagation technology, and further, a great amount of mulberry transgenic seedlings and seeds can be obtained. The method has the advantages that complicated steps such as tissue culture and requirements on sterile operation environment and the like are avoided; the operation is directly carried out on plants, and can be completed in natural environment; the cost is low; the operation is simple; the technical skill can be easily mastered; the repeatability is good; the conversion efficiency is higher; the transgenic plants, seedlings and seeds suitable for the local wild natural conditions can be fast obtained; the method is applicable to various mulberry varieties, and belongs to a novel efficient and practical mulberry transgenic method.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and relates to an agrobacterium-mediated transgenic method for mulberry trees. Background technique [0002] Woody plants are mostly perennial plants. Compared with crops and other annual and biennial herbaceous plants, they generally have the characteristics of long life cycle, late maturity and flowering, and complex genetic background. The research on their genetic transformation has been relatively lagging behind. In recent years, with the rapid development of Agrobacterium-mediated plant genetic transformation, researchers have successively studied poplar ( Populus L. ),locust( Robinia pseudoacacia Linn. ),apple( Malus pumila ),Peach( Amygdalus persica L. ),Tangerine( Citrus reticulata Blanco. ) and other woody plants successfully established a genetic transformation system. The genetic transformation of these woody plants is based on the characteristics of easy cultivation ...

AI Technical Summary

Problems solved by technology

Although some scholars have carried out transgenic research on mulberry trees through Agrobacterium-mediated leaf disc transformation and biolistic exogenous gene introduction and have made great progress, the leaf disc transformation method using leaves, cotyledons and other tissues is subject to strict and cumbersome tissue culture. Operational and difficult to establish mulberry regeneration systems are currently only successful in individual mulberry species (such as Indian mulberry and Shinichinose with strong regeneration ability), and these techniques are poorly reproducible and are currently only available in very few Several research groups have made breakthroughs, but it is still difficult to effectively apply them to functional genomics research and diversification development research of mulberry trees

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