Method for establishing influence of interacting protein on phytoplasma propagation through RNAi (Ribonucleic Acid Interference)

Abstract

The invention discloses a method for establishing influence of interacting protein on phytoplasma propagation through RNAi (Ribonucleic Acid Interference). The phytoplasma is wheat blue dwarf phytoplasma, the mediator is heterosachalina fulica, and the interaction protein is heterosachalina fulica microtubulin; the method comprises the following steps: synthesizing interaction protein genes dsRNA and dsGFP, respectively feeding a mediator with the interaction protein genes dsRNA and dsGFP, detecting the gene expression of the wheat blue dwarf phytoplasma by using qRT-PCR, infecting healthy wheat with the contaminated mediator to determine the virus transmission efficiency, and determining the influence of the interaction protein on phytoplasma transmission. According to the invention, the propagation influence of the heterosacharide microtubulin on the wheat blue dwarf phytoplasma is proved, and a foundation is laid for subsequent research on interaction of the phytoplasma and a mediator.

Description

technical field [0001] The invention belongs to the technical field of plant pathology, and in particular relates to a method for establishing the effect of interacting proteins on phytoplasma transmission through RNAi. Background technique [0002] Phytoplasma is a single-celled prokaryote that depends on vector insects for transmission and obligately parasitizes in the phloem of plants. It has no cell wall and belongs to Class Mollicutes, phylum Tenericutes. Usually, when a new phytoplasma disease is discovered, the identification of the transmission vector is mainly carried out on the insect species found near the diseased plants. Insects with organs are tested to determine whether they contain phytoplasma in their bodies. In the early stage, the phytoplasma in insect media was observed by electron microscope, and later with the development of PCR technology, the poisonous insects were determined by extracting insect DNA. [0003] Wheat blue dwarf disease (WBD) is one o...

AI Technical Summary

Problems solved by technology

Its piercing-sucking stylet can eat the stem and leaf vascular tissue of wheat plants, causing dry and yellow spots on the damaged parts, resulting in large-scale chlorosis of the tissue, affecting plant photosynthesis, and hindering plant growth and development

Vector leafhoppers can transmit various diseases such as viral diseases and phytoplasma. At present, wheat dwarf disease (WDV) and wheat blue dwarf disease (WBD) are more in-depth research. The occurrence and pandemic of diseases seriously affect wheat production and threaten wheat food security

[0005] Wheat blue dwarf disease is transmitted exclusively by the leafhopper, however, the effect of tubulin in the mediator, the interaction protein of the leafhopper, on the transmission of wheat blue dwarf disease is unclear

RNAi technology uses endogenous or exogenous dsRNA to mediate the specific degradation of homologous mRNA, so as to silence the expression of target genes, resulting in the loss of corresponding gene function

The introduction of dsRNA into insects is mainly through injection, tissue culture and feeding methods. Among them, the injection method is more common and its efficiency is high, but micro-injection is difficult, the mechanical damage is large, and it is easy to cause insect death. It is suitable for larger insects; Difficult to master basic conditions lead to difficult cell culture and slow development

Images