| Glycosyltransferase(GT,EC 2.4.xy)is responsible for the catalytic conversion of an enzyme in the glycosylation reaction,the most important transformation reaction in the biosynthesis of glycosylation,the glycosyltransferase in plants Glycosylation reaction can produce cascade effect,in order to change the plant compounds of various biological activity and function,thus affecting the growth and development of plants.In this study,the yeast-related glycosyltransferase GT43 B gene was obtained from Blast in NCBI,and the glycosyltransferase gene of rice was used as the research object.Jijing 88 as the recipient material,the use of modern bioinformatics means to find the rice glycosyltransferase subfamily GT43 and the subfamily were systematically analyzed.And to explore the relationship between the biological processes and molecular functions and the expression and interaction of different tissues and tissues in rice.On the basis of this,the GT43 B gene sequence in Jijing 88 was obtained,and the overexpression vector was successfully constructed.The transformation of Arabidopsis thaliana was carried out by Agrobacterium tumefaciens-mediated genetic transformation,and the function of the gene was verified.At the same time,rice callus was transformed to further clarify the function of the gene.The results are as follows:1.The rice glycosyltransferase subfamily GT43 consists of 12 members and belongs to the glucuronosyltransferase.The subfamily gene is distributed among multiple species and participates in multiple biological functions.Metabolic pathways,functional evolution.2.The relationship between the expression pattern of rice GT43 gene in different tissues,different periods and different hormone treatments was analyzed.The results showed that the expression of GT43 gene in rice was tissue-specific and spatiotemporal specificity,Such as GT43 B gene in the rice endosperm specific expression.According to the interaction network among the different members of the expression database,it is found that each member of the family cooperates with each other in the expression pattern.3.The GT43 B gene was successfully amplified and the overexpression vector was constructed.4.Agrobacterium tumefaciens-mediated genetic transformation was used to infect Arabidopsis thaliana.The T0 generation seeds were obtained by flower inserts of Arabidopsis thaliana,and positive Arabidopsis plants were successfully screened on the screening medium.The results showed that the transformed Arabidopsis thaliana was higher than the wild type chlorophyll level in the transformed Arabidopsis thaliana plant and wild type.The results showed that T2 generation of Arabidopsis thaliana was similar to that of wild type in the time of bolting time and flowering time.The results showed that the number of seeds,And the number of pods in T2 was increased compared with that of wild type,but the number of seeds in pods increased and the grain weight increased.5.The callus of rice was infected by Agrobacterium tumefaciens-mediated genetic transformation and the resistant callus was obtained.In the process of transformation,the callus infection time of rice was optimized.It was found that the transformation rate reached 17.32% when the infection was 20 min,and the rate of the long bacteria was 36.68%,although it was higher than the previous two groups but later The The results showed that callus was 4.74 times of callus before transformation and decreased with the increase of culture time.The content of soluble protein was found to be 1.85 times lower than that before transformation,and gradually increased with the increase of culture time.SOD activity was found to be 2.09 times higher than that of untransformed callus after transformation,and gradually decreased with the culture. |
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