机构地区:[1]长江大学园艺园林学院,湖北荆州434025 [2]武汉轻工大学国家富硒农产品加工技术研发专业中心,湖北武汉430023
出 处:《南方农业学报》2025年第1期135-148,共14页Journal of Southern Agriculture
基 金:国家自然科学基金项目(32371929);硒资源研究与生物应用湖北省重点实验室开放基金项目(PT10202311);恩施州“启航专项”科技计划项目(XYJ2023000008);荆州市科技局科技计划项目(2023EC40)。
摘 要:【目的】对构树bZIP基因家族成员进行鉴定及分析,并筛选构树中参与硒代谢的关键基因,为深入研究构树bZIP基因家族生物学功能及其在构树硒代谢中的作用机制提供理论依据。【方法】基于构树全基因组数据,利用生物信息学方法鉴定构树bZIP基因家族成员,并对其基因结构、启动子顺式作用元件、蛋白的系统进化关系等进行分析,并通过根癌农杆菌介导烟草叶片的瞬时转化法进行蛋白亚细胞定位。通过转录组测序(RNA-Seq)和实时荧光定量PCR分析构树bZIP家族基因在不同硒源及不同浓度处理下的表达水平。【结果】共鉴定得到38个bZIP基因家族成员(BpbZIP01~BpbZIP38),编码的氨基酸数量为140~894个,相对分子质量为16357.39~96969.63 Da,理论等电点为5.24~9.76,均为亲水性蛋白,不稳定系数为52.54~73.89,36个BpbZIPs蛋白定位于细胞核,BpbZIP15和BpbZIP19分别定位于过氧化物酶体和内质网。由系统发育进化树可知,将BpbZIPs蛋白分为10个亚族,BpbZIPs蛋白在各亚族中的数量为6个(A亚族)、1个(B亚族)、1个(C亚族)、7个(D亚族)、2个(E亚族)、1个(F亚族)、4个(G亚族)、2个(H亚族)、6个(I亚族)和8个(S亚族),各亚族中多数BpbZiPs与桑树bZIP聚成一支。不同BpbZIPs基因在外显子数量上差异较大,为1~16个,但相同亚族的基因长度和结构具有一定的相似性。BpbZIPs基因启动子含有光响应、激素和胁迫应答等多种响应元件。在不同浓度的硒酸钠和亚硒酸钠处理下,除BpbZIP17基因未表达外,其余BpbZIPs基因均对硒处理有上调或下调表达。结合相关分析和实时荧光定量PCR结果可知,亚硒酸钠和硒酸钠处理后BpbZIP04和BpbZIP05基因的表达水平均与总硒含量密切相关。【结论】构树bZIP基因家族成员参与多种生物和非生物胁迫,BpbZIP04和BpbZIP05基因在构树硒代谢与耐受性中发挥重要作用。【Objective】To identify and analyze the members of bZIP(basic leucine zipper)gene family members in Broussonetia papyrifera L.,and screen the key genes involved in selenium metabolism in B.papyrifera,which could provide theoretical reference for further study of the biological function mechanism of B.papyrifera bZIP gene family and its role in selenium metabolism.【Method】Based on the complete genome data of B.papyrifera,bioinformatics methods were used to identify the members of the bZIP gene family in B.papyrifera.The analysis included studying their gene structure,promoter cis-elements,protein systematic evolution relationships,and the transient transformation of tobacco leaves mediated by Agrobacterium tumefaciens was used for protein subcellular localization.The expression levels of B.papyrifera bZIP family genes under different selenium sources and concentrations were analyzed by transcriptome sequencing(RNASeq)and real-time fluorescence quantitative PCR.【Result】A total of 38 bZIP gene family members were identified(BpbZIP01-BpbZIP38),with number of amino acids encoded ranging from 140 to 894 amino acids,relative molecular weights ranging from 16357.39 to 96969.63 Da,theoretical isoelectric points ranging from 5.24 to 9.76,all of which were hydrophilic proteins,and instability coefficients ranging from 52.54 to 73.89.Among them,36 BpbZIPs were localized in the nucleus,while BpbZIP15 and BpbZIP19 were localized in the peroxisome and endoplasmic reticulum respectively.Based on the phylogenetic tree,the BpbZIPs proteins were divided into 10 subfamilies.The number of BpbZIPs proteins in each subfamily was 6(subfamily A),1(subfamily B),1(subfamily C),7(subfamily D),2(subfamily E),1(subfamily F),4(subfamily G),2(subfamily H),6(subfamily I)and 8(subfamily S).In each subfamily,most BpbZiPs proteins and the bZIP protein of the mulberry tree were clustered into a branch.The number of exons of different BpbZIPs genes varied greatly,ranging from 1 to 16,but the gene length and structure of the same subfamily
关 键 词:构树 bZIP基因家族 生物信息学分析 表达模式 硒代谢
分 类 号:S792.99[农业科学—林木遗传育种]
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