Cloning,Location and Expression Analysis of Gene StSN2 in Solanum tuberosum

doi:10.13560/j.cnki.biotech.bull.1985.2020-1198

Abstract

Abstract:

Snakin-2（StSN2）belongs to the Snakin/GASA family. GASAs play an important role in regulating plant growth and stress resistance. This work aims to clone the StSN2 gene,to study the function of StSN2 in the process of Solanum tuberosum tuber dormancy,and to explore the relationship between StSN2 and S. tuberosum tuber dormancy. The full-length CDS sequence of StSN2 was cloned from potato variety "Chuanyu 10" and analyzed by bioinformatics. The tissue expression specificity,dormancy release process and response to hormone were analyzed by qRT-PCR. The results showed that the open reading frame of the gene was 315 bp,encoding 104 amino acids,the relative molecular mass was about 11.04 kD,and the theoretical isoelectric point was 8.91;the protein was closely related to tomato,eggplant,tobacco and other Solanaceae plants in evolution. The results of tobacco transient expression revealed that the protein was localized in the nucleus and plasma membrane. Tissue expression analysis demonstrated that StSN2 was expressed in all tissues,with tubers being the highest,followed by leaves and roots,and lowest in stems,flower buds,and petioles. Further research discovered that the expression of StSN2 was regulated by ABA,BR and GA₃,especially in response to the induction of the dormant hormone ABA,and its expression decreased with the release of dormancy. In conclusion,StSN2 may be involved in hormone regulation of S. tuberosum dormancy.

Key words: Solanum tubersome, StSN2, tuber dormancy, gene cloning, expression analysis, hormone

PENG Jie, DENG Meng-sheng, ZHANG Jie, LIU Shi-feng, LUO Li-fei, WANG Xi-yao. Cloning,Location and Expression Analysis of Gene StSN2 in Solanum tuberosum[J]. Biotechnology Bulletin, 2021, 37(5): 11-18.

Figures/Tables 8

Fig. 1 Extraction of total RNA and amplification of StSN2 in S. tuberosum A: Potato total RNA electrophoresis. B: StSN2 amplification electrophoresis. C: colony PCR detection electrophoresis

Fig. 2 Coding sequence and amino acid sequence analysis of StSN2

Fig. 3 Prediction of secondary structure of StSN2 protein The purple part is irregularly curled, and the blue part is α-helix

Fig. 4 Prediction of tertiary structure of StSN2 protein A: StSN1 tertiary structure. B: StSN2 tertiary structure

Fig. 5 StSN2 phylogenetic tree

Fig. 6 Subcellular localization a: Fluorescence signal of fusion protein. b: Chloroplast fluorescence channel. c: Bright field. d: Superposition of bright field and fluorescence. e: No-load fluorescence signal. f: Chloroplast fluorescence channel. g: Bright field. h: Superposition of bright field and fluorescence

Fig. 7 Analysis of StSN2 expression in different tissues of S. tuberosun Different lowercase letters refer to significant differences (P<0.05)

Fig. 8 Germination rate and StSN2 expression analysis of S. tuberosun treated with three different hormones

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