盐度对波吉卵囊藻叶绿素荧光参数的影响及PsbA基因的克隆与表达分析  

Effects of Salinity on Chlorophyll Fluorescence Parameters and Cloning and Characterization of PsbA Gene in Oocystis Borgei

作　　者：任佳佳 洪廷 张宁 黄翔鹄[1,2,3] 李长玲 REN Jia-jia;HONG Ting;ZHANG Ning;HUANG Xiang-hu;LI Chang-ling(College of Fisheries,Guangdong Ocean University,Zhanjiang 524088,China;Shenzhen Research Institute,Guangdong Ocean University,Shenzhen 518000,China;Guangdong Research Center for Algae Culture and Applied Engineering,Zhanjiang 524025,China)

机构地区：[1]广东海洋大学水产学院,广东湛江524088 [2]广东海洋大学深圳研究院,广东深圳518000 [3]广东省藻类养殖及应用工程技术研究中心,广东湛江524025

出　　处：《广东海洋大学学报》2020年第3期30-39,共10页Journal of Guangdong Ocean University

基　　金：活性微藻制品产业化关键技术的研究和示范(KY20180112)。

摘　　要：【目的】研究波吉卵囊藻(Oocystis borgei)在不同盐度下叶绿素荧光参数的变化和编码叶绿体D1蛋白的PsbA基因的表达特性。【方法】利用叶绿素荧光仪测定不同盐度条件下波吉卵囊藻叶绿素荧光参数;利用cDNA末端快速扩增技术(RACE)克隆获取波吉卵囊藻PsbA基因序列,并进行生物信息学分析;利用实时荧光定量PCR(qPCR)方法分析PsbA基因在不同盐度下的表达情况。【结果】高盐(盐度60)条件下PSⅡ最大光化学量子产量(F_v/F_m)及PSII实际光化学量子效率(ΦPSⅡ)显著下降(P<0.05)。克隆获得1314 bp波吉卵囊藻PsbA基因cDNA序列,包含1062 bp开放阅读框(ORF),预测编码353个氨基酸,该蛋白属于跨膜蛋白。qPCR结果表明,高盐(盐度60)条件下PsbA基因极显著下调(P<0.01)。【结论】高盐(盐度60)显著抑制波吉卵囊藻PsbA基因的表达和叶绿素荧光参数(F_v/F_m和ΦPSⅡ)的降低,表明波吉卵囊藻处于胁迫状态;而盐度10和30能促进PsbA基因的表达。【Objective】To determine the effects of salinity on the changes of chlorophyll fluorescence parameters and expression characteristics of the PsbA gene encoding the chloroplast D1 protein of Oocystis borgei.【Method】Chlorophyll fluorescence meter was used to measure the chlorophyll fluorescence parameters of O.borgei under different salinity conditions.The rapid amplification of cDNA ends technology(RACE)was used to clone and obtain PsbA gene sequence.Its sequence characteristics were analyzed by bioinformatics.Real-time quantitative PCR(qPCR)was used to analyze the expression of PsbA under different salinity conditions.【Result】Under high salinity(salinity 60),the maximum photochemical quantum yield(F_v/F_m)of PSⅡand the quantum efficiency of electron transfer(ΦPSⅡ)of PSⅡdecreased significantly(P<0.05).The cDNA sequence of PsbA was 1314 bp in length.It contained an open reading frame(ORF)of 1062 bp and was predicted to encode a transmembrane protein composed of 353 amino acids.qPCR results showed that the PsbA was significantly down-regulated under high salinity conditions(salinity 60)(P<0.01).【Conclusion】The PsbA gene expression and chlorophyll fluorescence parameters(F_v/F_m andΦPSⅡ)were significantly inhibited at high salinity(salinity 60)when O.borgei was in stress;however,the salinity of 10 and 30 could promote expression of PsbA gene.

关 键 词：波吉卵囊藻 盐度 叶绿素荧光参数 PSBA基因 

分 类 号：S917.3[农业科学—水产科学]