Cloning of Mulberry MaERF105-Like Gene and Its Expression Under Drought Stress

doi:10.16420/j.issn.0513-353x.2021-0543

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (11): 2439-2448.doi: 10.16420/j.issn.0513-353x.2021-0543

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Cloning of Mulberry MaERF105-Like Gene and Its Expression Under Drought Stress

HUANG Renwei, REN Yinghong(), QI Weiliang, ZENG Rui, LIU Xinyu, DENG Binyan

College of Chemistry and Life Sciences，Sichuan Provincial Key Laboratory for Development and Utilization of Characteristic Horticultural Biological Resources，Chengdu Normal University，Chengdu 611130，China

Received:2022-06-08 Revised:2022-07-18 Online:2022-11-25 Published:2022-11-25
Contact: REN Yinghong E-mail:renyinghong@163.com

Abstract

Abstract:

MaERF105-Like gene and its promoter were cloned from‘Deguo 1’. Sequence analysis showed that the length of MaERF105-Like cDNA was 1 131 bp with an ORF of 882 bp，encoding 293 amino acids with molecular weight 32.43 kD and isoelectric point 8.71. MaERF105-Like has one AP2/ERF domain，belongs to B3 group of ERF family and shares a close relationship with tomato SlERF5. Subcellular localization prediction showed that MaERF105-Like might be located in nucleus. Promoter analysis showed that MaERF105-Like promoter possesses cis-acting elements involved in ABA，MeJA，ethylene and defense and stress responsiveness. qRT-PCR result showed that the expression level of MaERF105-Like was up-regulated by drought，and its expression was significantly down-regulated after rewatering treatment. The results of this study showed that MaERF105-Like might play an important role

in the resistance to drought in mulberry.

Key words: mulberry, MaERF105-Like, promoter, drought, expression analysis

CLC Number:

S663.9

HUANG Renwei, REN Yinghong, QI Weiliang, ZENG Rui, LIU Xinyu, DENG Binyan. Cloning of Mulberry MaERF105-Like Gene and Its Expression Under Drought Stress[J]. Acta Horticulturae Sinica, 2022, 49(11): 2439-2448.

Figures/Tables 6

References 27

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用途 Use	引物名称 Primer name	引物序列（5′-3′） Primer sequence	T_m/℃
基因克隆Gene cloning	MaERF105-Like-F	TCCTCTACTTTCGAACTTCTCAAC	57.1
	MaERF105-Like-R	CCTCAATCCCTTTGACTTTTCTTAT	59.9
启动子克隆Promoter cloning	pMaERF105-Like-F	CAACGGTATCACCCTCGCCAC	64.8
	pMaERF105-Like-R	TTCACTTCACCTTTACTCAAGTCCT	58.8
qRT-PCR	qMaERF105-Like-F	AAGGCGTTGAAGAGTTCGGTAG	60.6
	qMaERF105-Like-R	GAGATAACGGTGACAGTGGCG	60.4
	qMaβactin-F	TCCGATGCCCAGAAGTCCTC	62.1
	qMaβactin-R	TGATCTCCTTGCTCATCCTGTCT	60.4

用途 Use	引物名称 Primer name	引物序列（5′-3′） Primer sequence	T_m/℃
基因克隆Gene cloning	MaERF105-Like-F	TCCTCTACTTTCGAACTTCTCAAC	57.1
	MaERF105-Like-R	CCTCAATCCCTTTGACTTTTCTTAT	59.9
启动子克隆Promoter cloning	pMaERF105-Like-F	CAACGGTATCACCCTCGCCAC	64.8
	pMaERF105-Like-R	TTCACTTCACCTTTACTCAAGTCCT	58.8
qRT-PCR	qMaERF105-Like-F	AAGGCGTTGAAGAGTTCGGTAG	60.6
	qMaERF105-Like-R	GAGATAACGGTGACAGTGGCG	60.4
	qMaβactin-F	TCCGATGCCCAGAAGTCCTC	62.1
	qMaβactin-R	TGATCTCCTTGCTCATCCTGTCT	60.4

元件名称 Motif name	序列 Sequence	数量Number	功能 Function
A-box	CCGTCC	1	顺式调控元件 cis-Acting regulatory element
ABRE	ACGTG	2	脱落酸酸响应的顺式调控元件 cis-Acting element involved in the abscisic acid responsiveness
ARE	AAACCA	2	厌氧诱导必需元件 cis-Acting regulatory element essential for the anaerobic induction
ATC-motif	AGTAATCT	1	光应答中保守DNA模块的部分序列 Part of a conserved DNA module involved in light responsiveness
ATCT-motif	AATCTAATCC	1	光应答中保守DNA模块的部分序列 Part of a conserved DNA module involved in light responsiveness
Box 4	ATTAAT	2	光应答中保守DNA模块的部分序列 Part of a conserved DNA module involved in light responsiveness
CAAT-box	CAAT/CAAAT/CCAAT	28	启动子和增强子区域普通顺式作用元件 Common cis-Acting element in promoter and enhancer regions
CAT-box	GCCACT	1	与分生组织表达有关的顺式作用调控元件 cis-Acting regulatory element related to meristem expression
CGTCA-motif	CGTCA	1	茉莉酸甲酯响应的顺式调控元件 cis-Acting regulatory element involved in the MeJA-responsiveness
ERE	ATTTTAAA	4	乙烯响应元件 cis-Acting regulatory element involved in ethylene-responsiveness
G-Box	CACGTT	2	光响应元件 cis-Acting regulatory element involved in light responsiveness
MRE	AACCTAA	1	参与光响应的MYB结合位点 MYB binding site involved in light responsiveness
TATA-box	TATA/ATTATA/TATAA/TATAAATA/TATAAAT/TATAAA/TAAAGATT	26	启动子核心元件 Core promoter element around -30 of transcription start
TC-rich repeats	GTTTTCTTAC/ATTCTCTAAC	2	参与防御与胁迫应答的顺式调控元件 cis-Acting element involved in defense and stress responsiveness
TCT-motif	TCTTAC	1	部分光响应元件Part of a light responsive element
TGACG-motif	TGACG	1	茉莉酸甲酯响应的顺式调控元件 cis-Acting regulatory element involved in the MeJA-responsiveness

元件名称 Motif name	序列 Sequence	数量Number	功能 Function
A-box	CCGTCC	1	顺式调控元件 cis-Acting regulatory element
ABRE	ACGTG	2	脱落酸酸响应的顺式调控元件 cis-Acting element involved in the abscisic acid responsiveness
ARE	AAACCA	2	厌氧诱导必需元件 cis-Acting regulatory element essential for the anaerobic induction
ATC-motif	AGTAATCT	1	光应答中保守DNA模块的部分序列 Part of a conserved DNA module involved in light responsiveness
ATCT-motif	AATCTAATCC	1	光应答中保守DNA模块的部分序列 Part of a conserved DNA module involved in light responsiveness
Box 4	ATTAAT	2	光应答中保守DNA模块的部分序列 Part of a conserved DNA module involved in light responsiveness
CAAT-box	CAAT/CAAAT/CCAAT	28	启动子和增强子区域普通顺式作用元件 Common cis-Acting element in promoter and enhancer regions
CAT-box	GCCACT	1	与分生组织表达有关的顺式作用调控元件 cis-Acting regulatory element related to meristem expression
CGTCA-motif	CGTCA	1	茉莉酸甲酯响应的顺式调控元件 cis-Acting regulatory element involved in the MeJA-responsiveness
ERE	ATTTTAAA	4	乙烯响应元件 cis-Acting regulatory element involved in ethylene-responsiveness
G-Box	CACGTT	2	光响应元件 cis-Acting regulatory element involved in light responsiveness
MRE	AACCTAA	1	参与光响应的MYB结合位点 MYB binding site involved in light responsiveness
TATA-box	TATA/ATTATA/TATAA/TATAAATA/TATAAAT/TATAAA/TAAAGATT	26	启动子核心元件 Core promoter element around -30 of transcription start
TC-rich repeats	GTTTTCTTAC/ATTCTCTAAC	2	参与防御与胁迫应答的顺式调控元件 cis-Acting element involved in defense and stress responsiveness
TCT-motif	TCTTAC	1	部分光响应元件Part of a light responsive element
TGACG-motif	TGACG	1	茉莉酸甲酯响应的顺式调控元件 cis-Acting regulatory element involved in the MeJA-responsiveness

Cloning of Mulberry MaERF105-Like Gene and Its Expression Under Drought Stress

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