Rapid verification method for apple fruit size gene function

Abstract

A rapid verification method for an apple fruit size gene function comprises the steps of cloning of candidate genes, silencing of the candidate genes or construction of over expression vectors, infection of the candidate genes, phenotype analysis and gene identification of apple fruit samples, verification analysis of the candidate genes and the like. An instant function verification technology is adopted in the method, in a specific period of apple fruit development of a target sample, candidate genes are directly infected into apple fruits in an injection mode, target parts are directly infected by agrobacterium in a key period of fruit development, the purpose of quickly expressing and verifying gene functions is achieved, the problem that in the prior art, the fruit gene function verification period is long during a long apple juvenile period is solved, the problems of complex operation process and high verification cost in the prior art are solved, the verification period of the apple fruit gene function is shortened from 7-9 years to less than 1 year, the requirements of rapid and efficient verification of the fruit gene function in the current scientific research and production field of perennial crops such as apples are satisfied, the verification process is simplified, the verification cost is reduced, and the verification efficiency is improved.

Description

technical field [0001] The invention relates to a verification method of apple gene function, in particular to a rapid verification method of apple fruit size gene function. Background technique [0002] my country is the origin center of apple cultivation in the world, with rich apple germplasm resources and a wide range of cultivation areas. The cultivation area and output have always ranked first in the world for many years, and the annual output accounts for as much as 40% of the world's total annual output. To analyze the inheritance and molecular mechanism of apple germplasm, and use molecular biology techniques to regulate fruit size development and improve apple yield requires extensive and in-depth research on the gene functions of different varieties of apples. However, since apples are perennial crops, the growth period of seedlings is relatively long, and it usually takes 6 to 8 years to see fruit after sowing. Therefore, it takes at least 7 to 9 years to obtain ...

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Problems solved by technology

The traditional methods of transgenic function verification include cloning of candidate genes, transformation of Agrobacterium, co-cultivation of bacterial liquid and leaves, selection of stable transgenic plants on resistant medium, and verification of transgenic effects by qRT-PCR. The operation process is complicated and the verification cost is high. However, the test cycle is still long; moreover, there are still problems such as difficulty in obtaining stable transgenic systems and plants in apple varieties, and the verification of gene function of apple fruit size traits has adverse effects on obtaining accurate gene functions. Validation scientific research has brought great troubles and has become a limiting factor restricting scientific research on apple fruit

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