Expression patterns and transcriptional autoactivation analysis of CaERF70 in chili pepper

doi:10.3969/j.issn.1004-1524.20231318

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (10): 2247-2256.DOI: 10.3969/j.issn.1004-1524.20231318

• Horticultural Science • Previous Articles Next Articles

Expression patterns and transcriptional autoactivation analysis of CaERF70 in chili pepper

ZHANG Yu¹^,²^,³(), JIN Mingwei⁴, REN Li¹^,², ZHANG Yiying¹^,², ZHAO Hong¹^,², LIU Kun¹^,², DENG Shan¹^,², CHU Yunxia¹^,²^,³, LI Shouguo¹^,², ZHANG Jingli¹^,², HUANG Jingyan¹^,², CHEN Hairong¹^,²^,³^,^*()

1. Institute for Agri-Food Standards and Testing Technology, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
2. Shanghai Sub-Center for New Plant Variety Tests, Ministry of Agriculture and Rural Affairs, Shanghai 201415, China
3. Shanghai Key Lab of Protected Horticultural Technology, Horticultural Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
4. Agricultural College, AnShun University, AnShun 561000, Guizhou, China

Received:2023-11-21 Online:2024-10-25 Published:2024-10-30

Abstract

Abstract:

The AP2/ERF transcription factor family is widely involved in biological processes such as plant growth and development and stress response, so it is of great significance for the study of AP2/ERF transcription factors. In this study, we cloned a ERF gene, CaERF70 from salt-tolerant variety SHA2022119 of pepper, which contains 819 bp, encoding 272 amino acids, molecular weight of about 30.59 ku, and isoelectric point pI of 7.67. Bioinformatics analysis showed that the protein had a character with negatively charged, unstable, 34 potential phosphorylation sites, non-transmembrane and expressed in the nucleus. The phylogenetic tree results indicated that CaERF70 was clustered with the ERFs of tomato, tobacco and potato, suggesting that CaERF70 was very conserved in the evolution of Solanaceae. qRT-PCR showed that the expression of CaERF70 gene was induced by low temperature and suppressed by high temperature. Prokaryotic expression experiments showed that the size of the protein encoded by CaERF70 was about 30 ku consistent with the predicted size. Transcriptional activation activity analysis showed that CaERF70 had transcriptional activation activity. Taken together, the above results indicated that CaERF70 was an AP2/ERF2 transcription factor, belonging to the ERF Ⅶ group, with transcriptional activating activity, and the expression of CaERF70 was significantly changed due to high or low temperature stress, suggesting that CaERF70 was involved in the response of pepper to high or low temperature stress.

Key words: pepper, gene cloning, transcriptional activity, prokaryotic expression, abiotic stress

CLC Number:

S641.3

ZHANG Yu, JIN Mingwei, REN Li, ZHANG Yiying, ZHAO Hong, LIU Kun, DENG Shan, CHU Yunxia, LI Shouguo, ZHANG Jingli, HUANG Jingyan, CHEN Hairong. Expression patterns and transcriptional autoactivation analysis of CaERF70 in chili pepper[J]. Acta Agriculturae Zhejiangensis, 2024, 36(10): 2247-2256.

Figures/Tables 12

References 28

[1]	XIE Z L, NOLAN T M, JIANG H, et al. AP2/ERF transcription factor regulatory networks in hormone and abiotic stress responses in Arabidopsis[J]. Frontiers in Plant Science, 2019, 10: 228.
[2]	兰孟焦, 后猛, 肖满秋, 等. AP2/ERF转录因子参与植物次生代谢和逆境胁迫响应的研究进展[J]. 植物遗传资源学报, 2023, 24(5): 1223-1235.
	LAN M J, HOU M, XIAO M Q, et al. Research progress of AP2/ERF transcription factors participating in plant secondary metabolism and stress response[J]. Journal of Plant Genetic Resources, 2023, 24(5): 1223-1235. (in Chinese with English abstract)
[3]	SHARONI A M, NURUZZAMAN M, SATOH K, et al. Gene structures, classification and expression models of the AP2/EREBP transcription factor family in rice[J]. Plant and Cell Physiology, 2011, 52(2): 344-360.
[4]	ZHANG J, LIAO J Y, LING Q Q, et al. Genome-wide identification and expression profiling analysis of maize AP2/ERF superfamily genes reveal essential roles in abiotic stress tolerance[J]. BMC Genomics, 2022, 23(1): 125.
[5]	GHORBANI R, ZAKIPOUR Z, ALEMZADEH A, et al. Genome-wide analysis of AP2/ERF transcription factors family in Brassica napus[J]. Physiology and Molecular Biology of Plants, 2020, 26(7): 1463-1476.
[6]	WANG H T, NI D Q, SHEN J C, et al. Genome-wide identification of the AP2/ERF gene family and functional analysis of GmAP2/ERF144 for drought tolerance in soybean[J]. Frontiers in Plant Science, 2022, 13: 848766.
[7]	苟艳丽, 张乐, 郭欢, 等. 植物AP2/ERF类转录因子研究进展[J]. 草业科学, 2020, 37(6): 1150-1159.
	GOU Y L, ZHANG L, GUO H, et al. Research progress on the AP2/ERF transcription factor in plants[J]. Pratacultural Science, 2020, 37(6): 1150-1159. (in Chinese with English abstract)
[8]	NAKANO T, SUZUKI K, FUJIMURA T, et al. Genome-wide analysis of the ERF gene family in Arabidopsis and rice[J]. Plant Physiology, 2006, 140(2): 411-432.
[9]	AHARONI A, DIXIT S, JETTER R, et al. The SHINE clade of AP2 domain transcription factors activates wax biosynthesis, alters cuticle properties, and confers drought tolerance when overexpressed in Arabidopsis[J]. The Plant Cell, 2004, 16(9): 2463-2480.
[10]	BROUN P, POINDEXTER P, OSBORNE E, et al. WIN1, a transcriptional activator of epidermal wax accumulation in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(13): 4706-4711.
[11]	XU K N, XU X, FUKAO T, et al. Sub1A is an ethylene-response-factor-like gene that confers submergence tolerance to rice[J]. Nature, 2006, 442(7103): 705-708.
[12]	HATTORI Y, NAGAI K, FURUKAWA S, et al. The ethylene response factors SNORKEL1 and SNORKEL2 allow rice to adapt to deep water[J]. Nature, 2009, 460(7258): 1026-1030.
[13]	LEE D K, JUNG H, JANG G, et al. Overexpression of the OsERF71 transcription factor alters rice root structure and drought resistance[J]. Plant Physiology, 2016, 172(1): 575-588.
[14]	OH S J, KIM Y S, KWON C W, et al. Overexpression of the transcription factor AP37 in rice improves grain yield under drought conditions[J]. Plant Physiology, 2009, 150(3): 1368-1379.
[15]	LU J, JU H P, ZHOU G X, et al. An EAR-motif-containing ERF transcription factor affects herbivore-induced signaling, defense and resistance in rice[J]. The Plant Journal, 2011, 68(4): 583-596.
[16]	JIN J H, ZHANG H X, ALI M, et al. The CaAP2/ERF064 regulates dual functions in pepper: plant cell death and resistance to Phytophthora capsici[J]. Genes, 2019, 10(7): 541.
[17]	LEE J H, HONG J P, OH S K, et al. The ethylene-responsive factor like protein 1 (CaERFLP1) of hot pepper (Capsicum annuum L.) interacts in vitro with both GCC and DRE/CRT sequences with different binding affinities: possible biological roles of CaERFLP1 in response to pathogen infection and high salinity conditions in transgenic tobacco plants[J]. Plant Molecular Biology, 2004, 55(1): 61-81.
[18]	LAI Y, DANG F F, LIN J, et al. Overexpression of a pepper CaERF5 gene in tobacco plants enhances resistance to Ralstonia solanacearum infection[J]. Functional Plant Biology, 2014, 41(7): 758-767.
[19]	YOUM J W, JEON J H, CHOI D, et al. Ectopic expression of pepper CaPF1 in potato enhances multiple stresses tolerance and delays initiation of in vitro tuberization[J]. Planta, 2008, 228(4): 701-708.
[20]	TANG W, CHARLES T M, NEWTON R J. Overexpression of the pepper transcription factor CaPF1 in transgenic Virginia pine (Pinus virginiana Mill.) confers multiple stress tolerance and enhances organ growth[J]. Plant Molecular Biology, 2005, 59(4): 603-617.
[21]	TANG W, NEWTON R J, LI C, et al. Enhanced stress tolerance in transgenic pine expressing the pepper CaPF1 gene is associated with the polyamine biosynthesis[J]. Plant Cell Reports, 2007, 26(1): 115-124.
[22]	邹学校, 马艳青, 戴雄泽, 等. 辣椒在中国的传播与产业发展[J]. 园艺学报, 2020, 47(9): 1715-1726.
	ZOU X X, MA Y Q, DAI X Z, et al. Spread and industry development of pepper in China[J]. Acta Horticulturae Sinica, 2020, 47(9): 1715-1726. (in Chinese with English abstract)
[23]	JIN J H, WANG M, ZHANG H X, et al. Genome-wide identification of the AP2/ERF transcription factor family in pepper (Capsicum annuum L.)[J]. Genome, 2018, 61(9): 663-674.
[24]	赵慧, 杨诗勤, 徐凯, 等. 水稻耐旱性基因OsERF65的克隆、表达及转录自激活活性分析[J]. 分子植物育种, 2021, 19(2): 361-369.
	ZHAO H, YANG S Q, XU K, et al. Cloning,expression and trans-activation activity analysis of rice drought tolerance gene OsERF65[J]. Molecular Plant Breeding, 2021, 19(2): 361-369. (in Chinese with English abstract)
[25]	陈悦, 孙明哲, 贾博为, 等. 水稻AP2/ERF转录因子参与逆境胁迫应答的分子机制研究进展[J]. 作物学报, 2022, 48(4): 781-790.
	CHEN Y, SUN M Z, JIA B W, et al. Research progress regarding the function and mechanism of rice AP2/ERF transcription factor in stress response[J]. Acta Agronomica Sinica, 2022, 48(4): 781-790. (in Chinese with English abstract)
[26]	RIAZ M W, LU J, SHAH L, et al. Expansion and molecular characterization of AP2/ERF gene family in wheat (Triticum aestivum L.)[J]. Frontiers in Genetics, 2021, 12: 632155.
[27]	张余. 水稻抗逆相关基因OsEBP89和OsRMT1功能研究[D]. 武汉: 华中农业大学, 2020.
	ZHANG Y. Functional characterization of two stress related genes OsEBP89 and OsRMT1 in rice[D]. Wuhan: Huazhong Agricultural University, 2020. (in Chinese with English abstract)
[28]	雷雨晴, 张业猛, 王海庆. 扁蓿豆MrERF1的转录激活活性、亚细胞定位及表达分析[J]. 草业科学, 2021, 38(6): 1119-1127.
	LEI Y Q, ZHANG Y M, WANG H Q. Transcriptional activation, subcellular localization, and expression analysis of MrERF1 from Medicago ruthenica[J]. Pratacultural Science, 2021, 38(6): 1119-1127. (in Chinese with English abstract)

用途Usage	引物名称Primer name	引物序列Primer sequence
基因克隆Gene clone	CaERF70F	ATGGGTTCCCCACAAGAGAATTGCTC
	CaERF70R	TTATATCATGACAAGCTGAGAAT
表达量分析Expression analysis	RT-CaERF70F	GCCCCGAAAATACCCTTTATTC
	RT-CaERF70R	CGTTCGTTGAAACTCCTCTTTT
	RT-ActinF	AGGGATGGGTCAAAAGGATGC
	RT-ActinR	GAGACAACACCGCCTGAATAGC
原核表达Prokaryotic expression	GST-CaERF70F	TTCCAGGGGCCCCTGGGATCCATGGGTTCCCCACAAGAGAATTGCTC
	GST-CaERF70R	CTCGAGTCGACCCGGGAATTCTTATATCATGACAAGCTGAGAAT
自激活分析Self-activation analysis	BD-CaERF70F	ATGGCCATGGAGGCCGAATTCATGGGTTCCCCACAAGAGAATTGCTC
	BD-CaERF70R	CCGCTGCAGGTCGACGGATCCTTATATCATGACAAGCTGAGAAT

用途Usage	引物名称Primer name	引物序列Primer sequence
基因克隆Gene clone	CaERF70F	ATGGGTTCCCCACAAGAGAATTGCTC
	CaERF70R	TTATATCATGACAAGCTGAGAAT
表达量分析Expression analysis	RT-CaERF70F	GCCCCGAAAATACCCTTTATTC
	RT-CaERF70R	CGTTCGTTGAAACTCCTCTTTT
	RT-ActinF	AGGGATGGGTCAAAAGGATGC
	RT-ActinR	GAGACAACACCGCCTGAATAGC
原核表达Prokaryotic expression	GST-CaERF70F	TTCCAGGGGCCCCTGGGATCCATGGGTTCCCCACAAGAGAATTGCTC
	GST-CaERF70R	CTCGAGTCGACCCGGGAATTCTTATATCATGACAAGCTGAGAAT
自激活分析Self-activation analysis	BD-CaERF70F	ATGGCCATGGAGGCCGAATTCATGGGTTCCCCACAAGAGAATTGCTC
	BD-CaERF70R	CCGCTGCAGGTCGACGGATCCTTATATCATGACAAGCTGAGAAT

位置权重Location weight	LocDB	PotLocDB	Neural Nets	Pentamers	Integral
细胞核Nucleus	10.0	3.0	0	0.19	8.98
质膜Plasma membrane	0	0	0.96	0	0.68
细胞外Extracellular	0	0	0.96	0.19	0
细胞质Cytoplasm	0	0	0	2.42	0
线粒体Mitochondrion	0	0	0	1.88	0
内质网Endoplasmic reticulum	0	0	0	0.36	0
过氧化物酶体Peroxisome	0	0	0.96	0	0.06
高尔基体Golgi body	0	0	0.11	0.34	0
叶绿体Chloroplast	0	0	0	0.06	0.02
液泡Vacuole	0	0	0	0	0.26

位置权重Location weight	LocDB	PotLocDB	Neural Nets	Pentamers	Integral
细胞核Nucleus	10.0	3.0	0	0.19	8.98
质膜Plasma membrane	0	0	0.96	0	0.68
细胞外Extracellular	0	0	0.96	0.19	0
细胞质Cytoplasm	0	0	0	2.42	0
线粒体Mitochondrion	0	0	0	1.88	0
内质网Endoplasmic reticulum	0	0	0	0.36	0
过氧化物酶体Peroxisome	0	0	0.96	0	0.06
高尔基体Golgi body	0	0	0.11	0.34	0
叶绿体Chloroplast	0	0	0	0.06	0.02
液泡Vacuole	0	0	0	0	0.26

Expression patterns and transcriptional autoactivation analysis of CaERF70 in chili pepper

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