红苞凤梨AbF3'5'H基因上游调控因子筛选

doi:10.3969/j.issn.1004-1524.2023.01.09

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (1): 79-89.DOI: 10.3969/j.issn.1004-1524.2023.01.09

红苞凤梨AbF3'5'H基因上游调控因子筛选

林东璞(), 臧要强, 张霄鹏, 周徐子鑫, 马均()

四川农业大学风景园林学院,四川成都 611100

收稿日期:2022-03-28 出版日期:2023-01-25 发布日期:2023-02-21
通讯作者: *马均,E-mail: 422572009@qq.com
作者简介:林东璞(1998—),男,山东聊城人,硕士研究生,研究方向为观赏植物分子生物学。E-mail:373057871@qq.com
基金资助:
国家自然科学基金(31971704);国家自然科学基金(31770743)

Screening of upstream regulators of AbF3'5'H gene in Ananas comosus var. bracteatus

LIN Dongpu(), ZANG Yaoqiang, ZHANG Xiaopeng, ZHOU Xuzixin, MA Jun()

College of Landscape Architecture, Sichuan Agriculture University, Chengdu 611100, China

Received:2022-03-28 Online:2023-01-25 Published:2023-02-21

摘要/Abstract

摘要：

颜色是观赏植物最重要的性状之一,也是当前观赏园艺育种的一个重要方向。为研究红苞凤梨红蓝色彩的呈现机制,初步分析金边红苞凤梨(Ananas comosus var. bracteatus)不同组织中花青素种类与含量,用AbF3'5'H基因启动子构建诱饵载体,利用酵母单杂交(Y1H)文库筛选其上游调控转录因子;并采用酵母单杂交验证技术,验证筛选出的转录因子与AbF3'5'H启动子的互作关系。结果表明:花青素的种类组成和相对比例决定了组织的颜色性状,AbF3'5'H是花青素合成途径种类分支的关键基因,在花青素种类组成与相对比例调控中具有重要作用;AbBTB/POZ、AbHSP81-1和AbGLOX能够与AbF3'5'H的启动子结合而调控其转录,可能在红苞凤梨花青素合成代谢调控中发挥重要作用。研究结果对于进一步揭示红苞凤梨花青素合成代谢调控机理,研究红苞凤梨呈色机理,培育叶色新品种有着重要的理论和实践意义。

关键词: 红苞凤梨, F3'5'H基因, 花青素, 酵母单杂交, 调控因子

Abstract:

Color is one of the most important traits of ornamental plants and also an important direction of ornamental horticulture breeding. To investigate the presentation mechanism of red-blue color of Ananas comosus var. bracteatus, the primary analysis of the kinds and concentration of anthocyanins in different tissues had been made, the promoter of AbF3'5'H was used to build the bait vector, and the yeast one-hybrid (Y1H) library was used to select the transcription factors that control the gene’s upstream activity; The yeast one-hybrid technique was used to verify the interaction between the selected transcription factors and the AbF3'5'H promoter. The results showed that the phenotypic color of the tissues were determined by the kinds and relative proportion of anthocyanins. AbF3'5'H was a key gene functioned in the species branch determination of the anthocyanin synthesis pathway, and was crucial for regulating the kinds and relative proportion of anthocyanins; AbBTB/POZ, AbHSP81-1 and AbGLOX-like could bind to the promoter of AbF3'5'H to regulate its transcription, suggesting that they might play important role in the regulation of anthocyanin anabolism in A. comosus var. bracteatus. These results had important theoretical and practical significance in understanding the regulation mechanism of anthocyanin biosynthesis, investigating the color formation mechanism and cultivating new leaf color variants of A. comosus var. bracteatus.

Key words: Ananas comosus var. bracteatus, F3'5'H, yeast one-hybrid, regulatory factor

中图分类号:

S682.36

林东璞, 臧要强, 张霄鹏, 周徐子鑫, 马均. 红苞凤梨AbF3'5'H基因上游调控因子筛选[J]. 浙江农业学报, 2023, 35(1): 79-89.

LIN Dongpu, ZANG Yaoqiang, ZHANG Xiaopeng, ZHOU Xuzixin, MA Jun. Screening of upstream regulators of AbF3'5'H gene in Ananas comosus var. bracteatus[J]. Acta Agriculturae Zhejiangensis, 2023, 35(1): 79-89.

图/表 10

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引物名称 Primer name	引物序列 Primer sequence(5'→3')
AbF3'5'H-F	GGACGTTATCGTCCTCCTACG
AbF3'5'H-R	CGGCACAGTCTTTTGTAGCG
Pr F3'5'H-A-F	TAAGTCCATTTGAAACAGAA
Pr F3'5'H-A-R	TTTGTGAAGATTAAGAAGGC
Pr F3'5'H-B-F	TTAAAATTCAAATGACTGTCCAAAA
Pr F3'5'H-B-R	TTTTGGACAGTCATTTGAATTTTAA
F3'5'H-F	CCCGCATCATCATAACCAGC
F3'5'H-R	GCTTCAGGGAATCAGGGTGG

引物名称 Primer name	引物序列 Primer sequence(5'→3')
AbF3'5'H-F	GGACGTTATCGTCCTCCTACG
AbF3'5'H-R	CGGCACAGTCTTTTGTAGCG
Pr F3'5'H-A-F	TAAGTCCATTTGAAACAGAA
Pr F3'5'H-A-R	TTTGTGAAGATTAAGAAGGC
Pr F3'5'H-B-F	TTAAAATTCAAATGACTGTCCAAAA
Pr F3'5'H-B-R	TTTTGGACAGTCATTTGAATTTTAA
F3'5'H-F	CCCGCATCATCATAACCAGC
F3'5'H-R	GCTTCAGGGAATCAGGGTGG

顺式作用元件 Cis-acting elements	生物学功能 Biological function	出现次数 Emergency times
TATA-box	-30位置附近核心启动子转录起始位点Transcription start point-30 core promoter element	93
RY-element	种子特异性元件Seed specific element	1
ABRE	脱落酸响应元件Abscisic acid response element	8
LTR	低温响应元件Low temperature response element	3
MYB	MYB结合位点MYB binding site	5
AT1-motif	光响应的部分元件Some components that respond to light	2
CAAT-box	启动子和增强子共同作用顺式元件Regulatory elements for promoters and enhancers	10
Box 4	光响应的保守DNA模块的一部分Part of the conserved DNA module of the photoresponsive element	2
G-Box	参与光响应顺式元件Cis elements participating in photoresponse	5
TCA-element	水杨酸响应元件Salicylic acid response element	3
O2-site	醇溶蛋白代谢调节元件Cis-acting regulatory elements related to gliadin metabolism	1
GT1-motif	光响应元件Photoresponsive element	2
WUN-motif	创伤反应元件Wound response element	1
ARE	厌氧响应元件Anaerobic response element	9
circadian	昼夜节律控制调节元件Circadian rhythm control regulating element	1

顺式作用元件 Cis-acting elements	生物学功能 Biological function	出现次数 Emergency times
TATA-box	-30位置附近核心启动子转录起始位点Transcription start point-30 core promoter element	93
RY-element	种子特异性元件Seed specific element	1
ABRE	脱落酸响应元件Abscisic acid response element	8
LTR	低温响应元件Low temperature response element	3
MYB	MYB结合位点MYB binding site	5
AT1-motif	光响应的部分元件Some components that respond to light	2
CAAT-box	启动子和增强子共同作用顺式元件Regulatory elements for promoters and enhancers	10
Box 4	光响应的保守DNA模块的一部分Part of the conserved DNA module of the photoresponsive element	2
G-Box	参与光响应顺式元件Cis elements participating in photoresponse	5
TCA-element	水杨酸响应元件Salicylic acid response element	3
O2-site	醇溶蛋白代谢调节元件Cis-acting regulatory elements related to gliadin metabolism	1
GT1-motif	光响应元件Photoresponsive element	2
WUN-motif	创伤反应元件Wound response element	1
ARE	厌氧响应元件Anaerobic response element	9
circadian	昼夜节律控制调节元件Circadian rhythm control regulating element	1

红苞凤梨AbF3'5'H基因上游调控因子筛选

Screening of upstream regulators of AbF3'5'H gene in Ananas comosus var. bracteatus

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