Molecular identification method based on transfer of GAI mRNA molecules between pear rootstock and scion

Abstract

The invention discloses amino acid sequences of GAI proteins of Pyrus betulaefolia and Pyrus bretschneideri, and nucleotide sequences for coding the GAI proteins, wherein the sequences are disclosed as SEQ ID NO1-4 in the sequence table. The invention also discloses a molecular identification method based on transfer of GAI mRNA between pear rootstock and scion, which comprises the following steps: (1) micrografting seedlings in test tubes by using Pyrus bretschneideri as scion and using Pyrus betulaefolia as rootstock; (2) looking for the respective specific enzyme-digestion sites of the GAI genes of the Pyrus bretschneideri and the Pyrus betulaefolia, and respectively designing a primer at the upstream and downstream of the enzyme-digestion site; (3) and carrying out enzyme-digestion on the Pyrus betulaefolia, the Pyrus bretschneideri, and the RT-PCR product of the Pyrus betulaefolia and the Pyrus bretschneideri after grafting, and comparing the enzyme-digestion spectra before and after the grafting to identify the transfer conditions. The method is quick, sensitive, accurate, simple and convenient, and is applicable to other pear plants.

Description

technical field [0001] The invention relates to the field of plant molecular biology, in particular to a molecular identification method for the long-distance transfer of GAI gene mRNA molecules between rootstocks and scions of the genus Pyrus. Background technique [0002] In the production process of horticultural crops, grafting technology is widely used to obtain traits such as resistance and dwarfing, so as to increase yield and improve product quality. Among them, the application of grafting in fruit tree cultivation is more common. During the production process, it was found that the traits of the same scion grafted to different rootstocks are different. If the color, soluble solid content, flowering period, etc. change, the changes in many traits will affect the formation, growth and development of the fruit, and then affect the later stage. economic benefits. [0003] A series of studies have shown that some RNA molecules in plants are transferred between cells and...

AI Technical Summary

Problems solved by technology

[0005] So far, studies have found that there are many gene mRNAs and their encoded proteins that can be transmitted in the plant phloem over long distances, such as KN1, BEL5, NACP, FT protein, SUT1, etc. The verification of whether the RNA and proteins of these genes can be transmitted is also Only transgenic detection vectors carrying specific tags (Haywood et al., 2005), nested RT-PCR (Harada et al., 2009), Northern blot and other techniques are used. Although these techniques are relatively accurate in detecting gene transmissibility, However, the process is cumbersome, demanding, and time-consuming. There are obvious shortcomings such as heavy workload, long working cycle, and low efficiency. Especially because fruit tree transgenic and regeneration are difficult, we are studying the process of transferring RNA between rootstock and ear. , it is very difficult to verify the delivery of gene RNA by transgenic methods, which makes it even more difficult for us to carry out research on the grafting and delivery of endogenous mRNA in fruit trees

Images