尾叶桉叶绿体ROS代谢相关基因表达与不定芽分化的关系  

Relationship between Chloroplast ROS Metabolism Genes Expression and Adventitious Bud Differentiation in Eucalyptus urophylla

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作  者:杨玉红 方函 梁雪婷 黄真池 Yang Yuhong;Fang Han;Liang Xueting;Huang Zhenchi(Lingnan Normal University,Zhanjiang 524048,P.R.China;Hangzhou Normal University;Lingnan Normal University)

机构地区:[1]岭南师范学院,湛江524048 [2]杭州师范大学

出  处:《东北林业大学学报》2023年第9期41-45,共5页Journal of Northeast Forestry University

基  金:国家自然科学基金项目(31570660);湛江市科技发展专项资金竞争性分配项目(2022A01055);海南省果蔬贮藏与加工重点实验室项目(HNGS202105);国家级大学生创新创业项目。

摘  要:为研究不同类型愈伤组织的叶绿体活性氧(ROS)代谢相关基因表达,探讨尾叶桉再生型愈伤组织转绿和不定芽分化之间的关系,将9 d苗龄的尾叶桉(Eucalyptus urophylla S. T. Blake)幼苗下胚轴接种在添加N-苯基-N-噻唑基脲、6-苄氨基腺嘌呤、吲哚-3-乙酸的标准平板计数琼脂培养基(SPCA)中,外植体先膨大形成白色愈伤组织,8周后分化形成不同类型的愈伤组织(绿色带芽点愈伤组织、绿色愈伤组织、红色愈伤组织、白色愈伤组织)。4种愈伤组织中叶绿素质量分数分别为0.24、0.18、0.07、0.03 mg/g。实时荧光定量多聚核苷酸链式反应(qPCR)检测叶绿体rbcL基因和6个ROS代谢相关基因(TDP1、SOD2、DAR1、TRM4、TRF2、WTR1)的表达。qPCR检测到8周前的白色愈伤组织中4个叶绿体基因rbcL、TDP1、SOD2和DAR1的转录。其中,rbcL、TDP1、SOD2的转录水平随愈伤组织块的膨大不断升高,DAR1基因转录水平则较低。8周后,叶绿体rbcL基因和6个活性氧代谢相关基因均在绿色带芽点愈伤组织、绿色愈伤组织、红色愈伤组织中转录。设定红色愈伤组织中对应基因的表达量为1,绿色愈伤组织中,上述7个叶绿体基因(rbcL、TDP1、SOD2、DAR1、TRM4、TRF2、WTR1)相对表达量分别为8.84、3.36、2.89、2.65、2.53、1.28和7.49,绿色带芽点愈伤组织中7个基因的相对表达量分别为19.57、3.72、4.59、3.53、4.73、3.08和4.12。结果表明,愈伤组织转绿和不定芽分化过程中,叶绿体发育开始启动、叶绿素大量合成、rbcL转录显著增强,叶绿体中与活性氧代谢相关基因的表达水平升高。To investigate the relationship between organogenic calli turning green and adventitious shoot differentiation, we studied the expression of ROS metabolism related genes in chloroplasts of different types of calli. The hypocotyls of 9-d Eucalyptus urophylla S. T. Blake seedlingswere incubated on SPCa medium supplemented with PBU, 6-BA and IAA. Eight weeks later, the explants swelled and formed white callus. And the white callus differentiated into 4 types, including green callus with buds, green callus, red callus and white callus. The chlorophyll content of the four kinds of callus was 0.24, 0.18, 0.07 and 0.03 mg/g, respectively. The relative transcriptional levels of ribose-1, 5-bisphosphote carboxylase/oxygenase large subunit ( rbcL ) and six chloroplast genes related to ROS metabolism ( thioredoxin-dependent peroxidase 1 ( TDP1 ), Fe-superoxide dismutase 2 (SOD2 ), dehydroascorbate reductase 1 (DAR1 ), thioredoxin M-type 4 (TRM4), thioredoxin F2 (TRF2) and WCRKC thioredoxin 1(WTR1) ) were evaluated by qPCR. Four chloroplast genes, rbcL, TDP1, SOD2 and DAR1 were transcripted in 2, 4, 6 and 8-week white callus. The transcription levels of rbcL, TDP1 and SOD2 were increased with the expansion of callus, whilethe transcriptional level of DAR1 gene was lower. All 7 genes were transcripted in differentiated callus. By normalizing the transcriptional levels of corresponding genes in red callus to 1,the transcriptional levels of the seven genes ( rbcL, TDP1, SOD2, DAR1, TRM4, TRF2 and WTR1 ) in green callus were 8.84, 3.36, 2.89 , 2.65, 2.53, 1.28 and 7.49, respectively. The transcription levels of these seven genes in green callus with buds were 19.57, 3.72, 4.59, 3.53, 4.73, 3.08 and 4.12, respectively. The results implied that the development of chloroplast initiated during the differentiation of adventitious buds, which was accompanied with chlorophyll synthesizing, rbcL transcription increasing and the expression of chloroplast genes related to ROS metabolism enhancing.

关 键 词:尾叶桉 愈伤组织 叶绿体 基因表达 活性氧 

分 类 号:Q945.45[生物学—植物学]

 

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