Movable miRNA for promoting rootstock root development and drought resistance and identification method

Abstract

The invention belongs to the technical field of biotechnology and genetic improvement, and discloses a movable miRNA for promoting rootstock root development and drought resistance and an identification method. The movable miRNA for promoting rootstock root development and drought resistance is Mdm-miR160 in apple. The identification method of the movable miRNA for promoting rootstock root development and drought resistance comprises the following steps: RNA extraction and reverse transcription; gene cloning; genetic transformation of apples; drought stress treatment; in-situ hybridization; micro-grafting; tomato grafting on the tobaccos; and data analysis. The movable miRNA for promoting rootstock root development and drought resistance provided by the invention is Mdm-miR160 in apples, and can move from scions to rootstocks to promote root development of the rootstocks, so that the drought resistance of the apples is improved. An overexpressed transgenic line is obtained through genetic transformation of apples, and phenotypic analysis finds that Mdm-miR160e OE shows a dwarfing symptom.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and genetic improvement, and in particular relates to a mobile miRNA for promoting rootstock root system development and drought resistance and an identification method. Background technique [0002] Currently, plants have evolved dynamic and complex intercellular communication networks to coordinate and adapt their growth and development to various environmental changes. In addition to large molecules such as proteins and RNA, small molecules such as metabolites and plant hormones also play a role in signaling in plants. As a signal molecule, RNA can move locally between cells through plasmodesmata, or move long distances through phloem to exert biological functions. Non-cell autonomous RNAs can serve as mobile signals in various physiological processes such as plant growth and development, nutrient allocation, gene silencing, antiviral defense, and stress response, and have important biol...

AI Technical Summary

Problems solved by technology

[0006] Through the above analysis, the existing problems and defects of the prior art are as follows: 1. Most of the prior art is researched by sequencing analysis of phloem sap after grafting, in situ hybridization, stem-loop RT-qPCR, mutant screening and Northern blot. The movement of miRNAs, the above experiments can discover new mobile miRNAs, but there are few reports on the biological functions of miRNAs after movement

Second, miRNAs will be sheared twice during processing and generation, so fluorescent tags (such as GFP) cannot be attached to accurately visualize the movement of miRNAs

Third, the success rate of heterologous species grafting is low, and the miRNA family is very conserved among different species, which brings inconvenience to the experimental verification of miRNA movement

The signal transduction mechanism and regulation mechanism between cells and organs mediated by miRNA are less studied

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