Bioinformatics Analysis of BvM14-STPK Gene of Sugar Beet M14 Line and Expression Analysis of Its Response to Salt Stress

doi:10.11924/j.issn.1000-6850.casb20190700382

Abstract

Abstract:

We aim to study the BvM14-STPK protein kinase in response to salt stress. Sugar beet M14 line was used as material, the full length cDNA of BvM14-STPK gene was amplified by RT-PCR with specific primers. Bioinformatics was used to analyze the full-length cDNA sequence and real-time fluorescence quantitative PCR was carried out to determine the expression patterns of BvM14-STPK in the sugar beet M14 line in response to salt stress. The results of bioinformatics analysis showed that the full-length of BvM14-STPK had 1668 bp with a 1527 bp ORF, and encoded 508 amino acids. The BvM14-STPK contained a typical protein kinase conserved domain. The results of real-time fluorescence quantitative PCR showed that the BvM14-STPK gene was expressed in the leaves and roots, and the expression in leaves were higher than that of root. Under the 200 and 400 mmol/L NaCl stress, the gene showed different expression patterns in leaves and roots, indicating that the gene could answer salt stress, but the expressed quantity was different in different organs under salt stresses. This study is significant to improving quality genes of sugar beet M14 line and enhancing crop salt-stress tolerance, and could lay a foundation for further studying genetic engineering of sugar beet.

Key words: sugar beet M14 line, BvM14-STPK, bioinformatics analysis, fluorescence quantitative PCR, salt stress

CLC Number:

S566.3

Zhao Dongmei, Zhang Yongxue, Li Haiying. Bioinformatics Analysis of BvM14-STPK Gene of Sugar Beet M14 Line and Expression Analysis of Its Response to Salt Stress[J]. Chinese Agricultural Science Bulletin, 2020, 36(1): 35-42.

Figures/Tables 12

References 24

[1]	郭德栋, 康传红, 刘丽萍 . 异源三倍体甜菜(VVC)无融合生殖的研究[J]. 中国农业科学, 1999,32(40):1-5.
[2]	于冰, 李海英, 郭德栋 , 等. 甜菜无融合生殖系花期差异表达基因cDNA文库的构建[J]. 高技术通讯, 2006,16(9):954-957.
[3]	郭德栋, 刘丽萍, 康传红 , 等. 甜菜无融合生殖单体附加系的繁殖传递特性[J]. 黑龙江大学自然科学学报, 2001,18(3):104-107.
[4]	Li H, Pan Y, Zhang Y , et al. Salt stress response of membrane proteome of sugar beet monosomic addition line M14[J]. Journal of Proteomics, 2015,127:18-33. doi: 10.1016/j.jprot.2015.03.025 URL
[5]	Wu C, Ma C, Pan Y . Sugar beet M14 glyoxalaseⅠgene can enhance plant tolerance to abiotic stresses[J]. Journal of Plant Research, 2013,126(3):415-425. doi: 10.1007/s10265-012-0532-4 URL
[6]	Wang Y, Zhan Y, Wu C , et al. Cloning of a cystatin gene from sugar beet M14 that can enhance plant salt tolerance[J]. Plant Science, 2012,191-192(4):93-99. doi: 10.1016/j.plantsci.2012.05.001 URL
[7]	Zhu J . Abiotic stress signaling and responses in plants[J]. Cell, 2016,167(2):313-324. doi: 10.1016/j.cell.2016.08.029 URL
[8]	Sun X, Yu Q, Tang L , et al. GsSRK, a G-type lectin S-receptor-like serine/threonine protein kinase, is a positive regulator of plant tolerance to salt stress[J]. Journal of Plant Physiology, 2013,170(5):505-515. doi: 10.1016/j.jplph.2012.11.017 URL
[9]	Li J, Cai W , et al. A ginseng PgTIP1 gene whose protein biological activity related to Ser128 residue confers faster growth and enhanced salt stress tolerance in Arabidopsis[J]. Plant Science, 2015,234:74-85. doi: 10.1016/j.plantsci.2015.02.001 URL
[10]	Gong D, Guo Y, Jagendorf A T , et al. Biochemical characterization of the Arabidopsis protein kinase SOS2 that functions in salt tolerance[J]. Plant Physiology, 2002,130(1):256. doi: 10.1104/pp.004507 URL
[11]	裴丽丽, 郭玉华, 徐兆师 , 等. 植物逆境胁迫相关蛋白激酶的研究进展[J]. 西北植物学报, 2012,32(5):1052-1061.
[12]	朱婷婷, 王彦霞, 裴丽丽 , 等. 植物蛋白激酶与作物非生物胁迫抗性的研究[J]. 植物遗传资源学报, 2017,18(4):763-770.
[13]	Liu Z, Jia Y, Ding Y , et al. Plasma membrane CRPK1-mediated phosphorylation of 14-3-3 proteins induces their nuclear import to fine-tune CBF signaling during cold response[J]. Molecular Cell, 2017,66(1):117-128. doi: 10.1016/j.molcel.2017.02.016 URL
[14]	Yang L, Zhang Y, Zhu N , et al. Proteomic analysis of salt tolerance in sugar beet monosomic addition line M14[J]. Journal of Proteome Research, 2013,12(11):4931-4950. doi: 10.1021/pr400177m URL
[15]	端木慧子, 牛志新, 李海英 . 甜菜STPK家族鉴定及蛋白互作网络分析[J]. 中国糖料, 2018,40(06):8-10.
[16]	Meng L S, Wang Y B, Yao S Q , et al. Arabidopsis AINTEGUMENTA mediates salt tolerance by trans-repressing SCABP8[J]. Journal of Cell Science, 2015,128(15):2919. doi: 10.1242/jcs.172072 URL
[17]	Ma L, Ye J, Yang Y , et al. The SOS2-SCaBP8 complex generates and fine-tunes an AtANN4-dependent calcium signature under salt stress[J]. Developmental Cell, 2019,48(5):697-709. doi: 10.1016/j.devcel.2019.02.010 URL
[18]	Pareek A, Khurana A, Sharma A K , et al. An overview of signaling regulons during cold stress tolerance in plants[J]. Current Genomics, 2016,18(999).
[19]	Tomioka M, Shimobayashi M, Kitabatake M , et al. Ribosomal protein uS7/Rps5 serine-223 in protein kinase-mediated phosphorylation and ribosomal small subunit maturation[J]. Scientific Reports, 2018,8(1):1244. doi: 10.1038/s41598-018-19652-z URL
[20]	Liu J, Guo Y . The alkaline tolerance in Arabidopsis requires stabilizing microfilament partially through inactivation of PKS5 kinase[J]. Journal of Genetics and Genomics, 2011,38(7):307-313. doi: 10.1016/j.jgg.2011.05.006 URL
[21]	Huang K, Peng L, Liu Y , et al. Arabidopsis calcium-dependent protein kinase AtCPK1 plays a positive role in salt/drought-stress response[J]. Biochemical and Biophysical Research Communications, 2017,498(1).
[22]	Sarabi B, Fresneau C, Ghaderi N , et al. Stomatal and non-stomatal limitations are responsible in down-regulation of photosynjournal in melon plants grown under the saline condition: Application of carbon isotope discrimination as a reliable proxy[J]. Plant physiology and biochemistry, 2019,141:1-19. doi: 10.1016/j.plaphy.2019.05.010 URL
[23]	何亚飞, 李霞, 谢寅峰 . Rubisco与Rubisco活化酶的分子机理研究进展[J]. 分子植物育种, 2017,15(8):3295-3301.
[24]	陈候鸣, 陈跃, 王盾 , 等. 核酮糖-1,5-二磷酸羧化酶/加氧酶活化酶在植物抗逆性中的作用[J]. 植物生理学报, 2016,52(11):1637-1648.

引物名称	序列
S	5'-GGGTTTTGTTGAAGTGGCT-3'
AS	5'-AGCAAATTGTGATGGTCGAA-3'
YGS	5'-GTTACTGTGCTCCTGAATATG-3'
YGAS	5'-TGCCCGAGTGTTATCAATGG-3'
18S-F	5'-CCCCAATGGATCCGTTA-3'
18S-R	5'-TGACGGAGAATTAGGGTTCG-3'

引物名称	序列
S	5'-GGGTTTTGTTGAAGTGGCT-3'
AS	5'-AGCAAATTGTGATGGTCGAA-3'
YGS	5'-GTTACTGTGCTCCTGAATATG-3'
YGAS	5'-TGCCCGAGTGTTATCAATGG-3'
18S-F	5'-CCCCAATGGATCCGTTA-3'
18S-R	5'-TGACGGAGAATTAGGGTTCG-3'