Expression Analysis of Sl-miR482 in Tomato Fruit and the Construction of STTM Silencing Vector

doi:10.13560/j.cnki.biotech.bull.1985.2019-0566

Abstract

Abstract: The aim of this study is to investigate the specific expression pattern of tomato gene Sl-miR482 in fruit ripening stage,and to conduct bioinformatics analysis of its function and to construct STTM silencing vector. Quantitative PCR was used for gene expression detection,promoter analysis software and target gene prediction website was to perform bioinformatics analysis,PCR amplification,restriction enzyme digestion and ligation reaction were applied to construct STTM vector,and Agrobacterium was transformed. The expression level of Sl-miR482 increased gradually during flowering,and reached the peak in the immature green stage,and then gradually decreased. Further bioinformatics analysis showed that ethylene response elements were present in the Sl-miR482 promoter. The results of target gene prediction website showed that tomato pectin lyase Sl-PL13 was the target gene of Sl-miR482. Gene expression analysis showed that the expression levels of Sl-PL13 and Sl-miR482 demonstrated a trade-off relationship at different stages of fruit ripening. In sum,the STTM482 silencing vector is successfully constructed and transformed into Agrobacterium by PCR amplification and restriction enzyme ligation. The results of this study indicate that Sl-miR482 plays an important role in fruit ripening and softening.

Key words: Sl-miR482, STTM, fruit maturation and softening, regulatory mechanism, vector construction

MO Xian-lan, SHI Lie-qin, LU Qiu-li, WANG Xiao-min, REN Zhen-xin. Expression Analysis of Sl-miR482 in Tomato Fruit and the Construction of STTM Silencing Vector[J]. Biotechnology Bulletin, 2019, 35(12): 50-56.

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