Genetic transformation method of diploid populus tomentosa

Abstract

The invention relates to a genetic transformation method of diploid populus tomentosa. The genetic transformation method comprises the steps of firstly collecting tender bud-containing stem segments of male diploid populus tomentosa as explants, cleaning and sterilizing the segments, then sequentially carrying out propagation subculture, bud elongation culture and rooting culture to obtain asepticseedlings of populus tomentosa, picking agrobacterium tumefaciens carrying pBI121 vector plasmids, and preparing microbial liquid as infection liquid; cutting leaves of the aseptic seedlings of populus tomentosa into leaf discs, executing culturing for 7 days, then transferring the leaf discs into the infection liquid for infection, carrying out dark culture on the infected leaf discs, then carrying out light recovery culture, finally screening resistant buds to obtain candidate plants, carrying out GUS dyeing and PCR detection on the candidate plants, carrying out seedling hardening and transplantation on the plants which are all positive to obtain transgenic plants of diploid populus tomentosa. The method has the advantages of being high in conversion efficiency, short in cultivation period, capable of achieving large-scale production and the like, and the genetic transformation system of the diploid populus tomentosa can provide help for large-scale cultivation of new varieties anddirectional transformation of excellent characters.

Description

technical field [0001] The invention relates to a genetic transformation method of poplar tomentosa, in particular to a genetic transformation method of diploid poplar tomentosa, which belongs to the technical field of genetic engineering. Background technique [0002] Populus (Populus), as a general term for a class of plants in the family Salicaceae, is distributed in most parts of China. There are dozens of species of Populus in my country. Environmental adaptability and other characteristics. Populus is extremely rich in natural germplasm resources and is one of the most planted tree species in temperate climates in the world. Populus tomentosa (Populus tomentosa), as a native poplar species in China, has the characteristics of fast growth and strong environmental adaptability. The number of Populus tomentosa in my country is huge, and it is distributed in more than 10 provinces, cities and autonomous regions. Populus tomentosa grows rapidly, has strong adaptability, lo...

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Problems solved by technology

[0004] Different from other Populus plants, Populus tomentosa has a large number of wild triploid and diploid mixed plantings under natural conditions. Since triploid Populus tomentosa is taller in tree shape than diploid Populus tomentosa, it is a plant in some areas of my country. Shade trees and ornamental tree species are widely distributed, so the current genetic transformation system for Populus tomentosa is mostly aimed at triploid Populus tomentosa, and its regenerated plants have difficulty in rooting and developing, and it is difficult to form a complete plant, and the traditional Agrobacterium tumefaciens is usually used Mediated method, the efficiency of obtaining transgenic plants is generally less than 50% and most of them are false positives

[0005] At present, there are few related studies on the genetic transformation of diploid Populus tomentosa

As a native tree species in my country, diploid Populus tomentosa still has problems such as slow growth

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