Cloning and Characterization of Mugsp7, a β-1,3-glucanase Gene of Wild Banana Germplasm from Sanming, Fujian, China, Under Cold Treatment

doi:10.3969/j.issn.1000-2561.2020.02.012

Abstract

Abstract:

The tissue culture seedlings of a wild banana germplasm from Sanming were used to clone the cDNA and gDNA sequences of β-1,3-glucanase gene Mugsp7 by RT-PCR. Bioinformatics and qRT-PCR analysis of the expression at different low temperature were also carried out, and the enzyme activity of mperatureureent lowthe leaves was further determined by treating at 8 ℃ for 1, 2, 3, 4 and 5 days. The gDNA sequence of Mugsp7 was 1132 bp with an open reading frame (ORF) of 984 bp and one intron of 148 bp, which encoding 327 amino acids. Bioinformatics analysis showed that Mugsp7 was an acidic hydrophobic stable protein, and there was no signal peptide. The amino acid sequence of Mugsp7 shared significant similarity with that in Musa acuminata, Zostera marina, Maize, Hordeum vulgare L , Oryza sativa on the same branch, and it was closely related to Musa acuminata. The expression pattern of Mugsp7 under different low temperature treatments was different, suggesting that Mugsp7 was a low temperature stress related gene.

Key words: wild banana germplasm from Sanming, β-1,3-glucanase, Mugsp7, gene cloning, low temperature stress, qPCR

CLC Number:

Q785
Q789

XU Xiaoping,XIE Yanping,CHEN Fanglan,CHEN Xiaohui,CHEN Yukun,ZHANG Zihao,CHEN Chunzhen,LIN Yuling,LAI Zhongxiong. Cloning and Characterization of Mugsp7, a β-1,3-glucanase Gene of Wild Banana Germplasm from Sanming, Fujian, China, Under Cold Treatment[J]. Chinese Journal of Tropical Crops, 2020, 41(2): 292-299.

Figures/Tables 10

References 25

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生物信息学分析 Bioinformatics analysis	在线软件 Online s oftware	网址 Uniform resource locator
蛋白质基本理化性质	ProParam	http://web.expasy.org/protparam/
信号肽预测分析	SignalP 4.0 Server	http://www.cbs.dtu.dk/services/ SignalP/
亚细胞位点分析	PSORT	http://psort.hgc.jp/form.html
蛋白质跨膜结构分析	TMpred	http://www.ch.embnet.org/software/TMPRED_form.html
蛋白质保守区预测	NCBI-ProteinTools	http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi
蛋白质卷曲螺旋结构	Coils	http://www.ch.embnet.org/cgi-bin/COILS_form_parser
蛋白质二级结构	NPS	http://npsa-pbil.ibcp.fr
蛋白质三维结构	SEISS-MODEL	http://swissmodel.expasy.org/
磷酸化位点	NetPhos 2.0	www.predictprotein.org
同源性和分子系统进化	NCBI	http://blast.ncbi.nlm.nih.gov

生物信息学分析 Bioinformatics analysis	在线软件 Online s oftware	网址 Uniform resource locator
蛋白质基本理化性质	ProParam	http://web.expasy.org/protparam/
信号肽预测分析	SignalP 4.0 Server	http://www.cbs.dtu.dk/services/ SignalP/
亚细胞位点分析	PSORT	http://psort.hgc.jp/form.html
蛋白质跨膜结构分析	TMpred	http://www.ch.embnet.org/software/TMPRED_form.html
蛋白质保守区预测	NCBI-ProteinTools	http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi
蛋白质卷曲螺旋结构	Coils	http://www.ch.embnet.org/cgi-bin/COILS_form_parser
蛋白质二级结构	NPS	http://npsa-pbil.ibcp.fr
蛋白质三维结构	SEISS-MODEL	http://swissmodel.expasy.org/
磷酸化位点	NetPhos 2.0	www.predictprotein.org
同源性和分子系统进化	NCBI	http://blast.ncbi.nlm.nih.gov

引物名称 Primer name	引物序列 (5°-3°) Primer sequence (5°-3°)	退火温度 Annealing temperature/℃
Mugsp7-F	ATGGCATCAGAGTCTCGC	55
Mugsp7-R	TAGAAGTCCACGTGGTAGA	55
Mugsp7-qF	CATCTTCAGGGATTGGTGTC	60
Mugsp7-qR	GAAGGTGCCGAGGATGACT	60
18s-qF	CCTGAGAAACGGCTACCACAT	60
18s-qR	CACCAGACTTGCCCTCCA	60

引物名称 Primer name	引物序列 (5°-3°) Primer sequence (5°-3°)	退火温度 Annealing temperature/℃
Mugsp7-F	ATGGCATCAGAGTCTCGC	55
Mugsp7-R	TAGAAGTCCACGTGGTAGA	55
Mugsp7-qF	CATCTTCAGGGATTGGTGTC	60
Mugsp7-qR	GAAGGTGCCGAGGATGACT	60
18s-qF	CCTGAGAAACGGCTACCACAT	60
18s-qR	CACCAGACTTGCCCTCCA	60

	试管编号Test tube number
	1	2	3	4	5	6
含糖量/mg	0	0.4	0.8	1.2	1.6	2.0
葡萄糖/mL	0	0.4	0.8	1.2	1.6	2.0
蒸馏水/mL	2	1.6	1.2	0.8	0.4	0
DNS/mL	2	2	2	2	2	2