Identification of Auxin Receptor Gene TIR1 Family in Cyperus esculentus and the Expression Analysis in Response to Salt Stress and Exogenous IBA

doi:10.3969/j.issn.1000-2561.2023.05.003

Abstract

Abstract:

Cyperus esculentus is a special oil crop, with its underground tuber accumulating a large amount of oil, and also having the advantages of wide adaptability and strong stress resistance. This crop is a superior germplasm material for studying plant stress resistance and oil enrichment in the underground vegetative organs. Transport inhibitor response protein 1 (TIR1) can regulate plant growth and development and abiotic stress response as an auxin receptor. However, few reports on CeTIR1 have been reported. In this study, four TIR1 genes, namely CeTIR1-1, CeTIR1-2, CeTIR1-3 and CeTIR1-4, were identified from C. esculentus transcriptome data and the ORFs were further cloned by RT-PCR. All the four CeTIR1 proteins contained a typical F-box protein domain, AMN1 superdomain and Transp_inhibit domain, revealed by bioinformatics analysis. No significant difference was detected among the four CeTIR1 proteins in regarding sequence length, relative molecular weight, isoelectric point and other physicochemical properties. The four CeTIR1s were closely related to TIR1s derived from plants of Cyperaceae, Oleaceae and Cruciferae in the evolution. Gene expression profiling showed that the expression levels of CeTIR1 family genes were significantly different in different tissues of C. esculentus, with the highest expression in tuber tissues and lower expression in root and leaf tissues. The antioxidant enzyme activity and MDA content of C. esculentus seedlings increased under salt stress. After adding exogenous IBA treatment, the antioxidant enzyme activity of C. esculentus seedlings under salt stress further increased whereas the MDA content decreased. This indicated that IBA could alleviate the adverse effects of salt stress on the growth of C. esculentus seedlings. qRT-PCR results showed that expression levels of the four CeTIR1 genes increased in C. esculentus seedlings treated with salt stress, and exogenous addition of IBA could significantly up-regulate the expression of CeTIR1-2 gene. Comprehensive analysis demonstrates that CeTIR1-2 may be an important gene involved in regulating the response of C. esculentus seedlings to salt stress. Together, the results would provide the scientific basis and important reference for exploring the related function genes involved in auxin regulating the growth and development and stress resistance of C. esculentus, as well as the C. esculentus cultivation in the saline-alkali land.

Key words: Cyperus esculentus L., transport inhibitor response protein 1 (TIR1), indole butyric acid (IBA), salt stress, gene expression

CLC Number:

S565.9

LI Xinru, GAO Yu, MIAO Shunan, LI Teng, DONG Shuyan, SHI Xianfei, XUE Jin’ai, JI Chunli, LI Runzhi. Identification of Auxin Receptor Gene TIR1 Family in Cyperus esculentus and the Expression Analysis in Response to Salt Stress and Exogenous IBA[J]. Chinese Journal of Tropical Crops, 2023, 44(5): 894-904.

Figures/Tables 13

References 32

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引物 Primer	引物序列（5°-3°） Primer sequence (5°-3°)
ORF TIR1-1 F	ATGGCAGATGAACCGGCG
ORF TIR1-1 R	TTACAAGATCTTAACAAACGGCGG
ORF TIR1-2 F	ATGCGAGAGGAGATCTCTGACATG
ORF TIR1-2 R	TTACAAGATCTTAACAAAAGATGGTGC
ORF TIR1-3 F	ATGTCTGAGGAGGAGGAGGACG
ORF TIR1-3 R	CTACAATATCTTGACAAATTGCGGT
ORF TIR1-4 F	ATGAAGGTCCAATCGGCCA
ORF TIR1-4 R	TTACAGAGTCCATACAAAATCAGGTGC
qRT TIR1-1 F	CGACACGATTGGAGATGAAGG
qRT TIR1-1 R	GCGTTGACAGAAGAAGAGGATT
qRT TIR1-2 F	GCTCGGTCTCGCAGTGATT
qRT TIR1-2 R	GGCACAACCTACGCAATCG
qRT TIR1-3 F	GTGCCGTATCCTGTCAACCT
qRT TIR1-3 R	TCCAGTTCGTCATCCTCATTCA
qRT TIR1-4 F	TTCGCAGGAGACAGTGACC
qRT TIR1-4 R	TCCACATTCAACATCGGCATT
Ce18S rRNA F	CTACGTCCCTGCCCTTTGTACA
Ce18S rRNA R	ACACTTCACCGGACCATTCAA

引物 Primer	引物序列（5°-3°） Primer sequence (5°-3°)
ORF TIR1-1 F	ATGGCAGATGAACCGGCG
ORF TIR1-1 R	TTACAAGATCTTAACAAACGGCGG
ORF TIR1-2 F	ATGCGAGAGGAGATCTCTGACATG
ORF TIR1-2 R	TTACAAGATCTTAACAAAAGATGGTGC
ORF TIR1-3 F	ATGTCTGAGGAGGAGGAGGACG
ORF TIR1-3 R	CTACAATATCTTGACAAATTGCGGT
ORF TIR1-4 F	ATGAAGGTCCAATCGGCCA
ORF TIR1-4 R	TTACAGAGTCCATACAAAATCAGGTGC
qRT TIR1-1 F	CGACACGATTGGAGATGAAGG
qRT TIR1-1 R	GCGTTGACAGAAGAAGAGGATT
qRT TIR1-2 F	GCTCGGTCTCGCAGTGATT
qRT TIR1-2 R	GGCACAACCTACGCAATCG
qRT TIR1-3 F	GTGCCGTATCCTGTCAACCT
qRT TIR1-3 R	TCCAGTTCGTCATCCTCATTCA
qRT TIR1-4 F	TTCGCAGGAGACAGTGACC
qRT TIR1-4 R	TCCACATTCAACATCGGCATT
Ce18S rRNA F	CTACGTCCCTGCCCTTTGTACA
Ce18S rRNA R	ACACTTCACCGGACCATTCAA

植物 Plant	拉丁名 Latin name	蛋白 Protein	NCBI登录号 NCBI accession
大豆	Glycine max	GmTIR1	XP_006584933.1
蓖麻	Ricinus communis	RcTIR1	XP_015578180.1
花生	Arachis hypogaea	AhTIR1	XP_025689973.1
野生水稻	Oryza brachyantha	ObTIR1	XP_006647937.3
椰子	Cocos nucifera	CnTIR1	EHA8586401.1
康藏嵩草	Carex littledalei	ClTIR1	KAF3330046.1
油棕	Elaeis guineensis	EgTIR1	XP_010942647.1
柳枝稷	Panicum virgatum	PvTIR1	XP_039787433.1
油橄榄	Olea europaea	OeTIR1	CAA3002003.1
高梁	Sorghum bicolor	SbTIR1	XP_002465737.1
油菜	Brassica napus	BnTIR1	XP_022569407.1
穇子	Eleusine coracana	EcTIR1	GJN22350.1
黑麦草	Lolium rigidum	LrTIR1	XP_047075560.1

植物 Plant	拉丁名 Latin name	蛋白 Protein	NCBI登录号 NCBI accession
大豆	Glycine max	GmTIR1	XP_006584933.1
蓖麻	Ricinus communis	RcTIR1	XP_015578180.1
花生	Arachis hypogaea	AhTIR1	XP_025689973.1
野生水稻	Oryza brachyantha	ObTIR1	XP_006647937.3
椰子	Cocos nucifera	CnTIR1	EHA8586401.1
康藏嵩草	Carex littledalei	ClTIR1	KAF3330046.1
油棕	Elaeis guineensis	EgTIR1	XP_010942647.1
柳枝稷	Panicum virgatum	PvTIR1	XP_039787433.1
油橄榄	Olea europaea	OeTIR1	CAA3002003.1
高梁	Sorghum bicolor	SbTIR1	XP_002465737.1
油菜	Brassica napus	BnTIR1	XP_022569407.1
穇子	Eleusine coracana	EcTIR1	GJN22350.1
黑麦草	Lolium rigidum	LrTIR1	XP_047075560.1

蛋白 Protein	氨基酸 Amino acid	相对分子质量 Molecular weight/kDa	理论等电点Theoretical pI	不稳定系数Instability index	脂肪族指数Aliphatic index	亲水系数GRAVY	亚细胞定位Subcellalar localization
AtTIR1	594	66.80	6.99	47.58	94.83	0.009	细胞核
CeTIR1-1	589	65.67	5.13	42.64	91.88	-0.050	细胞核
CeTIR1-2	624	70.32	5.26	45.63	92.66	-0.080	细胞核
CeTIR1-3	611	67.11	6.10	46.33	98.00	0.017	细胞核
CeTIR1-4	599	66.37	6.72	45.34	94.06	-0.041	细胞核