Screening method of reference genes in stable expression in sitobion avenae fabricius at different temperatures and application

Abstract

The invention discloses a screening method of reference genes in stable expression in sitobion avenae fabricius at different temperatures and application and belongs to the field of molecular biology.According to the screening method, sitobion avenae fabricius treated at different temperatures is adopted as an experimental material to conduct fluorescence quantitative PCR (polymerase chain reaction) research, quantitative PCR data of each candidate reference gene is input into RefFinder for reference gene stability analysis, and thus reference gene-NADH (reduced form of nicotinamide-adenine dinucleotid) which is most suitable and stably expressed in the sitobion avenae fabricius in treatment at different temperatures can be screened. On the basis, the inventor takes the sitobion avenae fabricius after high temperature stress as a sample, takes a sitobion avenae fabricius heat shock protein hsp90 as a target gene, screen NADH genes as the reference genes, and studies expression rules of the sitobion avenae fabricius hsp90 genes at high temperature stress.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a screening method and application of an internal reference gene stably expressed at different temperatures of the plant aphid. Background technique [0002] Sitobion avenae Fabricius belongs to Homoptera and Aphididae. It is one of the main pests of wheat crops in my country. , sorghum, corn, sugar cane and other gramineous crops, as well as bluegrass, okra, crabgrass, clubhead grass, foxtail, white sheep grass and other gramineous and sedge weeds. It has the characteristics of wide distribution, large number and strong fecundity, which affects the normal growth of wheat plants, leads to a serious decline in wheat yield, and causes huge losses to agricultural production. In general years, wheat production can be reduced by 5.1% to 16.5%, and in major occurrence years, wheat production can be reduced by more than 40%. Generally, aphids are wingless, and can...

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Problems solved by technology

With the wide application of qRT-PCR technology, studies have found that, depending on the type of insect sample and experimental conditions, the expression level of housekeeping genes will change greatly under certain conditions. If you use it blindly, it may affect the final quantitative results. the accuracy of

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