A Preliminary Study On The Function Of BnWRKY16 And BnWRKY52 Transcription Factors

Rapeseed is the main oil crops in our country,to our country's economic development plays a decisive role.So,improving rapeseed seeds oil content of the research is the main target of rape breeding.WRKY transcription factors as control factor occupies a central position in the plant control network,involved in plant growth and development,stress resistance,disease resistance and so on a series of regulation.Some studies show that WRKY transcription factors may be involved in rape fat synthesis regulation.Therefore,this study used a set of oil content nearly isogenic line as the basic material,preliminary study of BnWRKY52?BnWRKY16 expression in the process of rape mature and germination,Specific circumstances are as follows:1.Based on the NJ9 high oil system as the base material,the protocol near-isogenic line were constructed using the method of continuous self.156 pairs of SSR premers from derived from Brassica DB public databases were used for PCR amplification of nine protocol near-isogenic line meterals,and 32 pairs of primers with clear and good repeatability were screened for near-isogenic analysis.The results of genetic similarity coefficient estimation and clustering analysis,based on PCR amplification result,showed that the genetic similarity coefficients of 9 candidate strains were between 0.94 and 0.85,and the genetic similarity coefficient of YC13-554 and YC13-559 was 0.94,which was identified as near-isogenic line.Based on the results of the investigation and analysis of Agronomic Traits in the mature period,the YC13-559 and YC13-554 were identified as high oil content and low oil near-iso genic line.2.The oil content difference of near isogenic line materials YC13-554 and YC13-559 as base material,and using the Quantitative Real-time PCR detect the gene expression of transcription factor BnWRKY52 and BnWRKY16 in the seed maturation process.The results showed that the expression of BnWRKY52 in YC13-554 and YC13-559 were increased,and the expression level in YC13-559 was higher than that in YC13-554;The expression of nWRKY16 in YC13-554 and YC13-559 only appeared a large peak,and the peak of expression in YC 13-559 was earlier than that in rape YC 13-554;According to the above results,it is speculated that transcription factor WKY52 and WRKY16 is related to the regulation of the synthesis of rapeseed oil.To verify the result,chose three identified key enzyme genes associated with rapeseed oil content:two acylglycerol acyltransferase(DTAG),WRI,CoA carboxylase,detect its expression,and make BnWRKY52 and BnWRKY16 expression and compare with them.The results showed that express similar between the two.3.By the oil content difference of near isogenic line material YC13-554 and YC 13-559 as base material,and using the Quantitative Real-time PCR detect the gene expression of transcription factor BnWRKY52 and BnWRKY16 in the seed germination process.Trizol method was used to extract material of RNA and reverse transcribed into cDNA,which used to detect transcription factor BnWRKY52,BnWRKY16 gene expression.Results show that BnWRKY16 and BnWRKY52 in rape YC 13-554 and rape YC 13-559 expression has two expression peaks,respectively,in seedlings through seed coat and long period of the first true leaf and in YC 13-559 than YC 13-554 starting in the expression of early.