坛紫菜转录因子NhbZIP1克隆和功能验证  

cDNA Cloning and Functional Verification of Transcription Factor NhbZIP1 from Neoporphyra haitanensis

作　　者：曾高雄 许凯[1,2,3] 徐燕 纪德华[1,2,3] 陈昌生 谢潮添[1,2,3] 王文磊 ZENG Gaoxiong;XU Kai;XU Yan;JI Dehua;CHEN Changsheng;XIE Chaotian;WANG Wenlei(Fisheries College,Jimei University,Xiamen,Fujian 361021,China;Fujian Engineering Research Center of Aquatic Breeding and Healthy Aquaculture,Xiamen,Fujian 361021,China;Key Laboratory of Healthy Mariculture for the East China Sea,Ministry of Agriculture and Rural Affairs,Xiamen,Fujian 361021,China)

机构地区：[1]集美大学水产学院,福建厦门361021 [2]福建省水产生物育种与健康养殖工程研究中心,福建厦门361021 [3]农业农村部东海海水健康与养殖重点实验室,福建厦门361021

出　　处：《渔业科学进展》2023年第1期201-209,共9页Progress in Fishery Sciences

基　　金：国家重点研发项目(2018YFD0901500);福建省重大科技专项(2019NZ08003);财政部和农业农村部:国家现代农业产业技术体系共同资助。

摘　　要：高温是制约坛紫菜(Neoporphyra haitanensis)产业发展的主要因素之一,阐明坛紫菜高温胁迫应答机理对耐高温品种选育至关重要。人们已分离多个坛紫菜抗逆相关基因,但尚不清楚这些基因的表达调控机制。本研究通过分子生物学和生物信息学技术分离了坛紫菜转录因子NhbZIP1基因。该基因开放阅读框长825 bp,编码274个氨基酸。从开放阅读框推导的氨基酸序列有5个低复杂度区域和1个BRLZ结构。其中,BRLZ是bZIP家族的保守结构域,含有一个α卷曲螺旋结构(121~171aa)。实时荧光定量PCR(qRT-PCR)检测发现,NhbZIP1受高温胁迫显著诱导。为进一步阐明NhbZIP1的功能,将其转入莱茵衣藻(Chlamydomonas reinhardtii)中。结果显示,高温胁迫下转基因藻株生物量始终高于野生型,且随处理时间增加差异越来越显著。转基因藻株中热激蛋白家族和抗氧化系统相关基因的表达量显著高于野生型。研究表明,NhbZIP1激活下游抗逆基因表达,在坛紫菜应答高温胁迫中发挥重要作用。研究结果有助于阐明bZIP调控坛紫菜响应高温胁迫的分子机制,为耐高温新品种选育提供了基础信息。Neoporphyra haitanensis is a macroalgae available in the south coast of China, and it is one of the most widely cultivated seaweeds in China. In recent years, due to global warming, the continuously high temperatures following the White Dew solar term, has led to the decomposition of seedlings of N. haitanensis in Fujian, Zhejiang and other provinces. This has had a huge impact on the coastal N. haitanensis cultivation industry in terms of production and development. Therefore,investigation of the molecular mechanism of high temperature stress response of N. haitanensis and the high-temperature resistance related genes is essential, and the results can also lay a foundation for the breeding of high-temperature resistant varieties of N. haitanensis. A previous study revealed that the basic region Leucine Zipper(bZIP) family transcription factors are one of the largest and most conserved transcription factor families in plants. The family plays an important role in plant response to abiotic stresses, such as high temperature, drought, and osmosis. bZIP transcription factors regulate plant response to abiotic stress by binding to functional genes or regulatory gene promoter cis-elements to activate and induce downstream gene expression. So far, 127, 89, and 216 bZIP transcription factors have been found in Arabidopsis, rice, and maize, respectively. However,previous studies on bZIP have mainly focused on model plants and only some field crops, and the functions of bZIP in macroalgae have not been reported. To this end, NhbZIP1 was screened based on the whole genome and transcriptomic data of N. haitanensis, and the NhbZIP1 gene was cloned and functionally analyzed by molecular biology and bioinformatics techniques. Its structure and expression pattern were also analyzed. Finally, the NhbZIP1 gene was transformed into Chlamydomonas reinhardtii by the “glass bead transformation” method for gene function verification.In this study, a gene product with a length of approximately 1000 bp was obtained by PCR amplificat

关 键 词：坛紫菜 高温胁迫 bZIP转录因子 转基因 莱茵衣藻 

分 类 号：S968.431[农业科学—水产养殖]