Study on Stress-related Function of Wheat E3 Ubiquitin Ligase TaSINA101

doi:10.7668/hbnxb.20195704

Abstract

Abstract:

The ubiquitination pathway is one of the key signaling pathways in response to drought stress.In order to clarify the biological function of E3 ubiquitin ligase TaSINA101 gene in response to drought stress,the TaSINA61,TaSINA101 and TaSINA105 genes were cloned from JM47,an excellent drought-resistant wheat cultivar,and their sequence characteristics were analyzed by bioinformatics methods,and the expression levels of the three genes in wheat roots and leaves were detected by qRT-PCR under PEG-6000,NaCl,low temperature and ABA treatments.The heterologous expression of TaSINA101 in transgenic rice was used to analyze the biological function of TaSINA101 in response to drought stress.The results showed that the TaSINA61,TaSINA101 and TaSINA105 genes contained one intron and two exons,and the encoded proteins were composed of 282 amino acids.The qRT-PCR expression analysis showed that the expression of these three genes was induced by various abiotic stresses such as drought stress in roots and leaves.Phenotypic analysis of TaSINA101 transgenic rice under drought stress showed that the leaf fresh weight and dry weight, maximum root length, average root diameter and leaf relative water content of transgenic rice lines OE-1, OE-2 and OE-3 were significantly lower than those of wild type,while the relative conductivity of leaves of transgenic rice lines OE-1 and OE-2 was significantly higher than that of wild type.Therefore,TaSINA101 negatively regulates drought stress tolerance in rice.This study provides a basis for in-depth analysis of the biological function of TaSINA101 gene in wheat.

Key words: Wheat, Drought stress, E3 ubiquitin ligase, Expression analysis, Functional studies, TaSINA101

CLC Number:

LIU Yaxin, CHEN Tao, GAO Weidong, GUO Lijian, CHE Zhuo, YANG Delong. Study on Stress-related Function of Wheat E3 Ubiquitin Ligase TaSINA101[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(4): 19-28. doi: 10.7668/hbnxb.20195704.

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Figures/Tables 8

Tab.1 Gene cloning and fluorescent quantitative PCR primer sequence

引物名称 Primer name	引物序列(5'-3') Primer sequence (5'-3')
TaSINA61-F	ATGGACGGGGGCACTACTAG
TaSINA61-R	TACACTGCGTACTGGTTTCAGAC
TaSINA101-F	ATGGATGGTGCTACCTCTAGCAAG
TaSINA101-R	TACACCGCGTATTGGTTTCAG
TaSINA105-F	ATGGATTGTGGTACCTCTAGCAAG
TaSINA105-R	TACACCGCGTACTGGTTTCAG
qRT-TaSINA61-F	GGCAAGAGGCAGAATGCAAC
qRT-TaSINA61-R	CGCTCGGGCATTCCTTCT
qRT-TaSINA101-F	CGAAGCTCATCGTTGTCTGA
qRT-TaSINA101-R	TACACCGCGTATTGGTTTCA
qRT-TaSINA105-F	TTAAGTTCACCCTAGAACCAGG
qRT-TaSINA105-R	GGGCTCATGCACCTTTACTTTAG
TaACTIN-F	GCCGTGCTTTCCCTCTATG
TaACTIN-R	GCTTCTCCTTGATGTCCCTTA

Fig.1 Cloning of TaSINA61, TaSINA101, and TaSINA105 genes M.DL 2000 DNA Marker; 1. TaSINA61 target fragment;2.TaSINA101 target fragment;3.TaSINA105 target fragment.

Fig.2 Sequence alignment of TaSINA61, TaSINA101 and TaSINA105 genes

Fig.3 Analysis of TaSINA61,TaSINA101 and TaSINA105 gene sequence and protein structure A:Schematic diagram of TaSINA61, TaSINA101 and TaSINA105 gene structures; B:Schematic diagram of TaSINA61,TaSINA101 and TaSINA105 domain prediction; C:The red rectangular box.RING-Ubox superfamily domain, The green rectangular box.SINA domain.

Tab.2 Gene information and protein physicochemical properties analysis

基因 Gene	基因号 Gene ID number	物理位置/bp Physical position	分子质量/ku Molecular weight	等电点 pI	蛋白长度/aa Protein length
TaSINA61	TraesCS3A02G030600	16 620 265-16 622 563	31.251 87	8.21	282
TaSINA101	TraesCS3B02G052800	26 746 166-26 748 742	31.435 24	8.03	282
TaSINA105	TraesCS3D02G029000	10 458 906-10 461 617	31.308 19	8.34	282

Fig.4 The cis-acting element of the TaSINA61,TaSINA101 and TaSINA105 promoter

Fig.5 Expression patterns of TaSINA61,TaSINA101 and TaSINA105 genes under salt, drought, low temperature and ABA treatment Different letters represent significant difference at the 0.05 level(P<0.05).The same as Fig.5.

Fig.6 Phenotype of TaSINA101 overexpressed rice lines under drought stress

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