Method for gene gun-mediated genetic transformation of moso bamboos

Abstract

The invention discloses a method for gene gun-mediated genetic transformation of moso bamboos. The method includes the steps: moso bamboo mature embryo acquisition and pre-culture; extraction of plasmid containing target genes; gene gun parameter setting; resistance screening; transformed plant identification; positive plant culture; transplantation. The method further includes a formula of a special culture medium in processes of transformation and induction. The method solves the problem of difficulty in genetic transformation of the moso bamboos, the period of acquiring transgenic bamboos can be effectively shortened (resistant seedlings are acquired after screening is performed for one month, and positive plants can be acquired for two to four months), and the probability of acquiring the transgenic bamboos is increased. The method is good in repeatability, plays an important role in industrial bamboo genetic improvement, bamboo function genomics research and the like, and has a wide application prospect.

Description

technical field [0001] The invention relates to a method for genetically transforming moso bamboo by means of a gene gun method, and belongs to the technical field of plant transgenics. Background technique [0002] Bamboo is a perennial evergreen plant in the subfamily Bambusoideae of Gramimineae. There are more than 70 genera and 1,250 species of bamboo resources in the world. There are 39 genera and more than 500 species of bamboo resources in my country. One of the forest resources, but also an important renewable resource. There are two main methods for improving the quality of bamboo fiber: one is the conventional breeding method, but due to the uncertainty of bamboo flowering, and the characteristics of death after flowering, there are huge restrictions; the other is through genetic engineering at the molecular level. Improve it. However, so far, the superiority of bamboo fiber has been verified mainly through some physical and chemical analysis, but no in-depth res...

AI Technical Summary

Problems solved by technology

The genotypes of Phyllostachys pubescens seeds vary greatly, and the callus induction rate and quality of callus of different genotypes of Phyllostachys pubescens are also quite different, and the repeatability is not good. At present, there are only a small number of reports about the induction and regeneration of Phyllostachys pubescens callus; and Agrobacterium bacteriostatic agents such as carbenicillin and cephalosporins are commonly used in the process of screening and cultivating, which can cause great damage to the callus of Phyllostachys pubescens, and no successful genetic transformation of Phyllostachys pubescens has been reported so far.

[0005] The Agrobacterium-mediated method is mainly used in dicotyledonous plants, but there are many restrictions on the application of monocotyledonous plants. The main problem is that Agrobacterium mainly infects dicotyledonous plants and very few monocotyledonous plants. Poaceae plants are not sensitive, and most of the most important food and economic crops in the world are Poaceae monocots, which limits its scope of application

In addition, the success of Agrobacterium-mediated genetic transformation is also highly dependent on the genotype. Even in dicotyledonous plants, many species are difficult to be transformed by Agrobacterium due to genotype restrictions.

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