Cloning and Expression Analysis of CsTMFs Gene in Tea Plant

doi:10.13560/j.cnki.biotech.bull.1985.2022-1408

Abstract

Abstract:

Based on the whole genome data of tea plant（Camellia sinensis）, members of tea plant TATA element modulatory factor（CsTMF）gene family were screened and identified. The full-length coding sequence（CDS）of CsTMFs in the tea plant leaves was further cloned. The conserved domain, gene structure, physical and chemical properties, protein secondary structure and phylogenetic relationship of CsTMFs were analyzed by bioinformatics method. Tissue-specific expression was analyzed based on transcriptome data, and qPCR was used to detect the expressions of CsTMFs in the tea plant leaves under cold stress and drought stress. The result showed that there were two genes of CsTMF gene family in the tea plant. The lengths of CDS region in two CsTMF genes were 2 934 bp and 2 904 bp, respectively. There were complete conserved domains, TMF-DNA-bd and TMF-TATA-bd in both two CsTMF genes. CsTMF1 and CsTMF2 had tissue expression specificity. CsTMF1 was highly expressed in the flowers and stems, while CsTMF2 was highly expressed in the mature leaves and stems. The results of qPCR showed that CsTMF1 was induced by cold stress, while CsTMF2 was inhibited by cold stress and drought stress.

Key words: tea plant, TMF, gene cloning, expression analysis

SUN Ming-hui, WU Qiong, LIU Dan-dan, JIAO Xiao-yu, WANG Wen-jie. Cloning and Expression Analysis of CsTMFs Gene in Tea Plant[J]. Biotechnology Bulletin, 2023, 39(7): 151-159.

Figures/Tables 13

References 20

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用途Application	引物名称Primer name	引物序列Primer sequence（5'-3'）
实时荧光定量PCR qPCR	CsTMF1-qF	TCCATCCGTGACTCTCTTGCTGAG
	CsTMF1-qR	GCCTCCGCCTTAGTGCCTCTA
	CsTMF2-qF	CGACTGGCATCTCTGGAATCC
	CsTMF2-qR	GCCTCCGTCTTAATGCTTCTAACT
	Actin-qF	GCCATCTTTGATTGGAATGG
	Actin-qR	GGTGCCACAACCTTGATCTT
基因克隆Gene cloning	CsTMF1-F	ATGGCCTGGTTTAGCGGGAAAGTCTCTTTGG
	CsTMF1-R	TCATGCAGAGCCCATAGAAGAACCCATTACC
	CsTMF2-F	ATGGCCTGGTTCGGTGGGAAAGTCT
	CsTMF2-R	CTAAGACGAACTTAGTACCTGGATC

用途Application	引物名称Primer name	引物序列Primer sequence（5'-3'）
实时荧光定量PCR qPCR	CsTMF1-qF	TCCATCCGTGACTCTCTTGCTGAG
	CsTMF1-qR	GCCTCCGCCTTAGTGCCTCTA
	CsTMF2-qF	CGACTGGCATCTCTGGAATCC
	CsTMF2-qR	GCCTCCGTCTTAATGCTTCTAACT
	Actin-qF	GCCATCTTTGATTGGAATGG
	Actin-qR	GGTGCCACAACCTTGATCTT
基因克隆Gene cloning	CsTMF1-F	ATGGCCTGGTTTAGCGGGAAAGTCTCTTTGG
	CsTMF1-R	TCATGCAGAGCCCATAGAAGAACCCATTACC
	CsTMF2-F	ATGGCCTGGTTCGGTGGGAAAGTCT
	CsTMF2-R	CTAAGACGAACTTAGTACCTGGATC

Motif	宽度Width	最可能序列Most possible match	结构域Domain
1	80	TARLQPSTVPDQSPITRRKSGFENGNLSRKLSSASSLGSMEESFFLQASLDSSDNFSERRNPGEMAMSPYFMKSMPPSAF	*
2	78	TMLVQALEELRQTLSRKEQQAVFREDMLRRDIEDLQKRYQESERRCEELVTQVPESTRPLLRQIEAMQETTGRRAEAW	*
3	40	MAWFGGKVSLGNFPDLAGAVNKISESVKNIEKNFDSALGF	*
4	80	MKMMETALQGAARQAQAKADEIAKLMNENENQKAVIEDLRRKSNEAEIESLREEYHQKVAALERKVYALTKERDTLRREQ	*
5	80	IRDSLAEELVKMTAQCEKLRVEAAILPGIRAELEALRRRHSAALELMGERDEELEELRADIVDLKEMYREQVNLLVNKIQ	TMF-TATA-bd
6	79	GLWPSGTDRKTJFEPVIAFMGHKGGESTVESIEKPELLEPPSSVEEKZEGENDRSTDFATEQTRPAEEVNZESSHMLPD	*
7	79	QAGMEVELPGFGFPTSKEIESAEEHFTDDLPDGPPPDEAAEMVSEPVSHENDTFGREVELNZQASDYEPDIKEQRVSSG	TMF-DNA-bd
8	80	VMAEGEELSKKQAAQESQIRKLRAQIRELEEEKKGLTTKLQVEENKVESIKRDKAETEKLLQETIEKHQAEJAVQKEFYT	*
9	80	QTLSRLNVLEAQISCLRAEQTQLSRSLEKERQRAAENRQEYLAAKEEADTHEGRVSQLEEEIRELRRKQKZELQEALMHR	*
10	66	TSALGPFENSELVEAKPGTNEVDQVEVGAPVPPESHNVINLHEISDEQKAZEEEIVENLSPVQAED	*

Motif	宽度Width	最可能序列Most possible match	结构域Domain
1	80	TARLQPSTVPDQSPITRRKSGFENGNLSRKLSSASSLGSMEESFFLQASLDSSDNFSERRNPGEMAMSPYFMKSMPPSAF	*
2	78	TMLVQALEELRQTLSRKEQQAVFREDMLRRDIEDLQKRYQESERRCEELVTQVPESTRPLLRQIEAMQETTGRRAEAW	*
3	40	MAWFGGKVSLGNFPDLAGAVNKISESVKNIEKNFDSALGF	*
4	80	MKMMETALQGAARQAQAKADEIAKLMNENENQKAVIEDLRRKSNEAEIESLREEYHQKVAALERKVYALTKERDTLRREQ	*
5	80	IRDSLAEELVKMTAQCEKLRVEAAILPGIRAELEALRRRHSAALELMGERDEELEELRADIVDLKEMYREQVNLLVNKIQ	TMF-TATA-bd
6	79	GLWPSGTDRKTJFEPVIAFMGHKGGESTVESIEKPELLEPPSSVEEKZEGENDRSTDFATEQTRPAEEVNZESSHMLPD	*
7	79	QAGMEVELPGFGFPTSKEIESAEEHFTDDLPDGPPPDEAAEMVSEPVSHENDTFGREVELNZQASDYEPDIKEQRVSSG	TMF-DNA-bd
8	80	VMAEGEELSKKQAAQESQIRKLRAQIRELEEEKKGLTTKLQVEENKVESIKRDKAETEKLLQETIEKHQAEJAVQKEFYT	*
9	80	QTLSRLNVLEAQISCLRAEQTQLSRSLEKERQRAAENRQEYLAAKEEADTHEGRVSQLEEEIRELRRKQKZELQEALMHR	*
10	66	TSALGPFENSELVEAKPGTNEVDQVEVGAPVPPESHNVINLHEISDEQKAZEEEIVENLSPVQAED	*

物种名Species name	登录号Accession No.
水稻Oryza sativa Japonica Group	BAG90311.1
茭白Zizania palustris	KAG8086397
小麦Triticum aestivum	XP 044346450.1
大麦Hordeum vulgare subsp. Vulgar	BAJ99499
玉米Zea mays	XP 008656031.1