The Research Of The Correlation Between Ver2 Gene And Albinism Leaves In Wheat Albinism Line

Wheat albinism line is a low-temperature-induced albinism natural mutation and it was found during the process of studying its parent(Wheat Aibian 1)temperature and regulated by nuclear genes and cytoplasmic genes.In recent years,many studies of its genetics,physiology and cell have been conducted.In this research,Wheat albinism line and its parent Wheat Aibian 1 were both treated with low temperature,Ver2 gene was indentified as the Wheat albinism line albinism related genes by studying their gene chips and digital expression profiling.Ver2 gene is related with the wheat vernalization phenomenon,which is a low-temperature induced gene and encodes a class of plant lectin protein,only expressed in the heart leaf tissue and surrounding young leaves.Wheat albinism line is a kind of special material that was albinism from heart leaf under low temperature.So,we selected Ver2 gene as candidate to explore the molecular mechanisms of low-temperature induced albinism of Wheat albinism line.In our study,Ver2 gene DNA and CDNA sequence were cloned from Wheat albinism line,and Ver2 gene promoter was cloned by chromosome walking techniques,through which we analyzed the molecular characteristics of Ver2 gene and try to find out the albinism related experimental basis in nucleic acid level.In order to find out the relationship between Ver2 gene at the transcriptional level and albinism phenomenon,the expression difference of Ver2 gene at the transcriptional level was detected under different tissues and conditions.The prokaryotic and eukaryotic expression vectors of Ver2 gene were constructed to research the correlation between protein level and albinism phenomenon,and Ver2 gene function validation.For the Ver2 gene could affect the times of spike differentiation and heading,we have made a comparison of them between Wheat albinism line and wheat Aibian 1 for further detection of Ver2 gene function.The main conclusions are as follows:1、By sequence alignment,we found that the Ver2 gene cDNA we cloned was 903bp long,which was same as the published ones by GenBank.And Ver2 gene we cloned were 1471 bp long,which was same as the gene sequence published from Jingdong No.1.2、The Ver2 gene promoter was cloned by Chromosome walking technique.When compared with the published Ver2 gene promoter,there was a deletion from-270 to-1891 in upstream of the gene,many important elements included.3、According to semi-quantitative research in albinism,albinism sterile material and its parent,we found that Ver2 gene only expressed in albinism heart leaf under room temperature.And under low temperature,it only expresses in the materials with albinism phenomenon.4、According to the detection of Ver2 gene DNA in several materials,we found that Ver2 gene was also amplified and transcribed under room temperature in Jinmai 47,Xiaoyan No.6,Xiaoyan No.22,Zhangye,Gansu,Luoyang 8716.5、Prokaryotic expression vector Ver2-pET28a and eukaryotic expression vectors pUC18-35s::Ver2-eGFP of Ver2 gene were built,and the prokaryotic expression of Ver2 protein was successfully induced.6、The spike development of wheat albinism line and wheat Aibian 1 was compared,from which we found that both of them were of normal spike development,but the spike development of wheat albinism line was earlier than that of wheat Aibian 1.Also,the heading time of wheat albinism line was 5 days earlier than that of wheat Aibian 1,the panicle length and grain weight of the two materials were significantly different(P<0.05),the panicle length of wheat albinism line was 7.8%longer than that of wheat Aibian 1 by 7.8%,the kernel weight wheat Aibian 1 was heavier than that of wheat albinism line by 2.9%.In conclusion,further research on the correlation between Ver2 gene and albinism leaves is necessary,and we think the deletion sequence of Ver2 promoter may affect its expression.