RT-PCR Analysis And Identification Of Anthocyanin Synthesis Related Genes In Triticum Aestivum L.cv.Guizi1 Seed

As a special kind of purple wheat germplasm resources,its peel contains a lot of anthocyanins,anthocyanins in health care,medical,food and so on various aspects all play an important role,but to participate in the regulation of anthocyanin biosynthesis network is very complicated,which not only have anthocyanins genes involved in,plant hormones may also be involved in some of the related gene regulation of anthocyanin,therefore,studying these genes played a role in anthocyanin synthesis is very necessary.The main research object of this study was GZ1 wheat.The expression levels of anthocyanin-related genes in GZ1 and Chinese spring were analyzed.The contents of auxin and cytokinin in different periods(10dpa,25 dpa,35dpa)and antioxidant enzyme in the accumulation of anthocyanin in GZ1 seeds were determined.Based on the previous transcriptome data,the heat maps of auxin and cytokinin-related genes were analyzed,and the related genes were verified by q RT-PCR.The GzANS-6D gene was cloned and transformed into tobacco.1.In this study,the expression of 8 anthocyanin-related genes(GzMYB-7D1 and GzMYC-2A1 are anthocyanin regulatory genes,GzCHI-5D ? GzANS ? GzCHS ?GzPAL-6D1-T2?GzDFR-3B-T1 and GzF3H-2D are structural genes)in GZ1 and Chinese spring different periods(10dpa,25 dpa,35dpa)in were detected.q RT-PCR results showed that the expression levels of 8 anthocyanin-related genes in GZ1 were all higher than that in Chinese spring,but these 8 genes showed different expression patterns in GZ1 and Chinese spring.The results showed that the expression patterns of GzMYB-7D1 ? GzPAL-6D1-T2 and GzCHI-5D were consistent with the accumulation patterns of anthocyanin,the expression patterns of GzMYC-2A1?GzANS and GzCHS were similar to the accumulation patterns of anthocyanin,and the expression patterns of GzDFR-3B-T1 and GzF3H-2D were opposite to the accumulation patterns of anthocyanin.Presumably GzMYB-7D1 and GzMYC-2A1 your purple wheat involved in regulation no.1,the synthesis of anthocyanin GzCHS,GzPAL-6D1-T2 and GzCHI-5D with your purple wheat grain color may have a direct relationship,GzANS gene is highly expressed in GZ1,which is nearly 200 times higher than other genes.Therefore,it is believed that GzANS plays an important role in anthocyanin accumulation.GzDFR-3B-T1 and GzF3H-2D may have no direct effect on the grain color of GZ1 wheat.2.This study of auxin and cytokinin in your GZ1 wheat grain the role of anthocyanin synthesis analysis,the auxin and cytokinin content detection results show that in your GZ1 wheat in different periods of seed growth(10dpa,25 dpa and 35dpa),auxin content change trend of decline after rising first,25 dpain anthocyanin synthesis is an important period,reached the highest content of auxin and cytokinin content is the basic trend of gradually reduce.We analyzed the heat maps and expression levels of auxin and cytokinin-related genes,and found that the expression levels of auxin polarity transport vectors(GzPIN1 and GzLAX3),auxin receptors(GzABP and GzARP)and signaling elements(most of GzARFs and GzAUX/IAAs)at 25 dpa and 35 dpa were significantly higher than those at 10 dpa,with an increasing and then decreasing pattern.Both the cytokinin-degrading gene(GzCKX)and the inactivation gene(GzUGT)gradually decreased,which means that the degradation or inactivation of endogenous cytokinin was gradually eliminated.These results indicate that cytokinin play a negative regulatory role in the accumulation of anthocyanin in GZ1 grain.Auxin and cytokinin may play an antagonistic role in anthocyanin accumulation by regulating related genes during seed development of GZ1 wheat.The detection of reactive oxygen species found that reactive oxygen signals were also involved in the regulation of anthocyanin accumulation in GZ1.3.The GzANS-6D gene was cloned from GZ1,and the overexpression vector of GzANS-6D-p BI121 was constructed and transformed into agrobacterium root nosa.Anthocyanins of tobacco related genes in q RT-PCR analysis,the results of the study have found GzANS-6D express basic improve other structural gene expression in the tobacco,and the gene specificity in GZ1 wheat high expression,thus we speculated that GzANS-6D is GZ1 wheat grain color the dominant phenotypic differences is the most crucial factor,but it is not the only key genes.The results of this study laid a foundation for auxin and cytokinin to participate in the synthesis of anthocyanin in GZ1,and provided a reference for the in-depth study on the function of GZ1 GzANS-6D in anthocyanin synthesis.