猕猴桃后熟过程中ABA响应结合因子AcAREB1调控AcGH3.1的表达

doi:10.16420/j.issn.0513-353x.2021-1086

摘要/Abstract

摘要：

以‘红阳’猕猴桃为试材，研究脱落酸（ABA）对果实采后成熟过程中生长素（IAA）代谢的影响。结果表明，外源ABA处理能够加速自由态IAA的降解，提高IAA-Asp含量，同时可以显著促进IAA降解相关基因AcGH3.1的表达。从猕猴桃基因组数据库中分离鉴定到5个ABA响应结合因子基因（AcAREB1 ~ AcAREB5），荧光定量PCR显示，5个基因在果实采后成熟过程中均能不同程度响应ABA。酵母单杂交结果显示，AcAREB1能够直接结合到AcGH3.1的启动子上。亚细胞定位显示AcAREB1定位在细胞核上。酵母自激活试验表明AcAREB1具有转录激活活性。这些结果表明，在猕猴桃果实成熟过程中，AcAREB1可以响应ABA，调控AcGH3.1的表达，从而促进IAA的降解，最终使得果实开始软化成熟。

关键词: 猕猴桃, 后熟, AcAREB1, AcGH3.1, 启动子, ABA, IAA

Abstract:

The effect of ABA on IAA metabolism during postharvest ripening of‘Hongyang’kiwifruit were studied. The results showed that exogenous ABA treatment could accelerate the degradation of free IAA，increase the content of IAA-Asp，and significantly promote the expression of IAA degradation related gene AcGH3.1. Five ABA-responsive binding factor genes（AcAREB1-AcAREB5）were isolated and identified from kiwifruit genome database. qRT-PCR showed that all five AcAREB genes could respond to ABA during postharvest ripening. Yeast one-hybrid results showed that AcAREB1 could directly bind to the AcGH3.1 gene promoter. Subcellular localization assay showed that AcAREB1 was located on the nucleus. Yeast self-activation experiments proved that AcAREB1 was a transcription factor with transcriptional activation activity. These results indicate that AcAREB1 can regulate the expression of AcGH3.1 gene corresponding to ABA during the ripening process of kiwifruit，thereby promoting the degradation of IAA，and finally leading to fruit softening and ripening.

Key words: kiwifruit, postharvest ripening, AcAREB1, AcGH3.1, promoter, ABA, IAA

中图分类号:

S663.4

袁馨, 徐云鹤, 张雨培, 单楠, 陈楚英, 万春鹏, 开文斌, 翟夏琬, 陈金印, 甘增宇. 猕猴桃后熟过程中ABA响应结合因子AcAREB1调控AcGH3.1的表达[J]. 园艺学报, 2023, 50(1): 53-64.

YUAN Xin, XU Yunhe, ZHANG Yupei, SHAN Nan, CHEN Chuying, WAN Chunpeng, KAI Wenbin, ZHAI Xiawan, CHEN Jinyin, GAN Zengyu. Studies on AcAREB1 Regulating the Expression of AcGH3.1 During Postharvest Ripening of Kiwifruit[J]. Acta Horticulturae Sinica, 2023, 50(1): 53-64.

图/表 11

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基因 Gene	上游序列（5′-3′）Forward sequence	下游序列（5′-3′）Reverse sequence
AcGH3.1	AGGGTCTGTACTTCCTGTTCG	TGTAGACGTTGTAGGGATCG
AcAREB1	GCAACAACAACCACCTTCAA	GTTCCCAAACCAATCATTCC
AcAREB2	GGGGCGGCAGTAGTGAACCA	AGTGTCAGACCAGGCGTAAT
AcAREB3	GGTCAGCGCACAGGATTGAG	AGCCAGATTTGATTGCTTGG
AcAREB4	ACGGATAGGAATAGTGCTGG	TAAATCACCGAAGAAGCCAC
AcAREB5	ATGGCTTCTTCGGTGATTTA	CTGCTGCTGTGATGATCTAAC
GUS	CACAAACCGTTCTACTTTACTGG	GCGTAAGGGTAATGCGAGGT
AcActin	TGGAATGGAAGCTGCAGGA	CACCACTGAGCACAATGTTGC

基因 Gene	上游序列（5′-3′）Forward sequence	下游序列（5′-3′）Reverse sequence
AcGH3.1	AGGGTCTGTACTTCCTGTTCG	TGTAGACGTTGTAGGGATCG
AcAREB1	GCAACAACAACCACCTTCAA	GTTCCCAAACCAATCATTCC
AcAREB2	GGGGCGGCAGTAGTGAACCA	AGTGTCAGACCAGGCGTAAT
AcAREB3	GGTCAGCGCACAGGATTGAG	AGCCAGATTTGATTGCTTGG
AcAREB4	ACGGATAGGAATAGTGCTGG	TAAATCACCGAAGAAGCCAC
AcAREB5	ATGGCTTCTTCGGTGATTTA	CTGCTGCTGTGATGATCTAAC
GUS	CACAAACCGTTCTACTTTACTGG	GCGTAAGGGTAATGCGAGGT
AcActin	TGGAATGGAAGCTGCAGGA	CACCACTGAGCACAATGTTGC

引物名称 Primer name	引物序列（5′-3′） Primer sequence
ProAcGH3.1-pHIS2	F:GGGCGAATTCCCGGGGAGCTCCTTTTATATATATATATATAT
	R:CGGATCGATTCGCGAACGCGTATATGAAGTGGTCCCCAATT
AcAREB1-BD	F:TCAGAGGAGGACCTGCATATGATGAACTTCAAGAACTATGG
	R:CCGCTGCAGGTCGACGGATCCTTAGCTCCTTGCTGCGTGTT
AcAREB1-AD	F:CGTACCAGATTACGCTCATATGATGAACTTCAAGAACTATGG R:GCAGCTCGAGCTCGATGGATCCTTAGCTCCTTGCTGCGTGTT
AcAREB2-AD	F:CGTACCAGATTACGCTCATATGATGGGATCATATTTGAACTT R:GCAGCTCGAGCTCGATGGATCCTTATTCATTGCATAAAATAG
AcAREB3-AD	F:CGTACCAGATTACGCTCATATGATGTACTTCAAGAACTATGG R:GCAGCTCGAGCTCGATGGATCCTCACCATGGGCTAGTCTGTG
AcAREB4-AD	F:CGTACCAGATTACGCTCATATGATGAACTTCAGAAACTTTGG R:GCAGCTCGAGCTCGATGGATCCTCATGTTTCAACTATTGAAT
AcAREB5-AD	F:CGTACCAGATTACGCTCATATGATGAACTTCAGAAACTTTGA R:GCAGCTCGAGCTCGATGGATCCTCACCATGGGCCAGTTTGTG
AcAREB1-GFP	F:CCAAATCGACTCTAGTCTAGAATGAACTTCAAGAACTATGG R:GCCCTTGCTCACCATGGTACCGCTCCTTGCTGCGTGTT
ProAcGH3.1-GUS	F:AACTGCAGCTTTTATATATATATATATAT R:CGGGATCCATATGAAGTGGTCCCCAATT

引物名称 Primer name	引物序列（5′-3′） Primer sequence
ProAcGH3.1-pHIS2	F:GGGCGAATTCCCGGGGAGCTCCTTTTATATATATATATATAT
	R:CGGATCGATTCGCGAACGCGTATATGAAGTGGTCCCCAATT
AcAREB1-BD	F:TCAGAGGAGGACCTGCATATGATGAACTTCAAGAACTATGG
	R:CCGCTGCAGGTCGACGGATCCTTAGCTCCTTGCTGCGTGTT
AcAREB1-AD	F:CGTACCAGATTACGCTCATATGATGAACTTCAAGAACTATGG R:GCAGCTCGAGCTCGATGGATCCTTAGCTCCTTGCTGCGTGTT
AcAREB2-AD	F:CGTACCAGATTACGCTCATATGATGGGATCATATTTGAACTT R:GCAGCTCGAGCTCGATGGATCCTTATTCATTGCATAAAATAG
AcAREB3-AD	F:CGTACCAGATTACGCTCATATGATGTACTTCAAGAACTATGG R:GCAGCTCGAGCTCGATGGATCCTCACCATGGGCTAGTCTGTG
AcAREB4-AD	F:CGTACCAGATTACGCTCATATGATGAACTTCAGAAACTTTGG R:GCAGCTCGAGCTCGATGGATCCTCATGTTTCAACTATTGAAT
AcAREB5-AD	F:CGTACCAGATTACGCTCATATGATGAACTTCAGAAACTTTGA R:GCAGCTCGAGCTCGATGGATCCTCACCATGGGCCAGTTTGTG
AcAREB1-GFP	F:CCAAATCGACTCTAGTCTAGAATGAACTTCAAGAACTATGG R:GCCCTTGCTCACCATGGTACCGCTCCTTGCTGCGTGTT
ProAcGH3.1-GUS	F:AACTGCAGCTTTTATATATATATATATAT R:CGGGATCCATATGAAGTGGTCCCCAATT

名称 Name	基因编号 Gene ID	染色体定位 Chromosome No.	蛋白长度/aa Protein length	分子量/kD Molecular weight	等电点 Isoelectric point	亚细胞定位 Subcellular localization
AcAREB1	Ach00g131571	未知 Unknow	431	47.30	9.79	细胞核 Nuclear
AcAREB2	Ach06g233651	06	448	49.08	9.16	细胞核 Nuclear
AcAREB3	Ach00g227711	未知 Unknow	411	45.04	10.10	细胞核 Nuclear
AcAREB4	Ach11g026271	11	459	49.43	5.40	细胞核 Nuclear
AcAREB5	Ach05g315671	05	396	42.63	7.51	细胞核 Nuclear