闽楠NRT/NPF基因家族鉴定与表达分析  

Identification and Expression Analysis of NRT/NPF Gene Family in Phoebe bournei

作　　者：李翼贝 童再康 韩双 王立 韩潇 张毓婷 杨琪 黄华宏 张俊红 LI Yi-Bei;TONG Zai-Kang;HAN Shuang;WANG Li;HAN Xiao;ZHANG Yu-Ting;YANG Qi;HUANG Hua-Hong;ZHANG Jun-Hong(School of Forestry&Bio-technology,Zhejiang A&F University/State Key Laboratory of Subtropical Silviculture,Hangzhou 311300,China)

机构地区：[1]浙江农林大学林业与生物技术学院/省部共建亚热带森林培育国家重点实验室,杭州311300

出　　处：《农业生物技术学报》2023年第10期2072-2086,共15页Journal of Agricultural Biotechnology

基　　金：浙江省农业新品种选育重大专项(2021C02070-10);浙江省重点研发计划项目(2021C02054)。

摘　　要：氮素作为植物不可或缺的大量元素,其含量显著影响植物生长发育,其中硝态氮为闽楠(Phoebe bournei)等诸多陆生植物氮素吸收的主要形式,对硝酸盐转运蛋白(nitrate transporter/peptide transporter family,NRT/NPF)基因家族鉴定与表达分析具重要意义。本研究利用拟南芥(Arabidopsis thaliana)NRT蛋白序列和隐马科夫模型(PF00854,PF07690和PF16974)在闽楠全基因组鉴定出63个NRT/NPF家族成员,不均匀分布于11条染色体,其分子量为22.8~135.6 kD,等电点为5.44~9.41,编码205~1235个氨基酸,其编码蛋白多为碱性蛋白,且含有2~23个跨膜结构域;系统进化分析将63个PbNRT/NPF分为3个亚家族,其中60个属于NRT1/NPF亚家族,2个属于NRT2亚家族,1个属于NRT3亚家族。共线性分析表明,PbNRT/NPF发生了9个串联重复事件和10个片段重复事件。转录组数据和qRT-PCR分析均表明,PbNRT/NPF成员在叶、根皮部、根木质部、茎皮部和茎木质部5种组织中的表达模式存在显著差异,其中硝酸盐转运蛋白基因PbNPF6.3在根皮部表达显著高于其他组织。qPCR结果表明,PbNRT/NPF成员在不同低氮处理阶段呈差异表达,其中PbNRT2.4和PbNRT2.5表达量在低氮处理下显著上升。本研究为进一步探索PbNRT/NPF家族成员在氮素吸收、转运和利用中的生物学功能提供了重要基础,为闽楠氮素高效利用新品种选育提供依据。Nitrogen is one of the essential macronutrient for plants,thus nitrogen content in soil seriously affects plant growth and development.Nitrate as the most important source of nitrogen absorption for P.bournei and many other land plants,and the nitrate transporter/peptide transporter family(NRT/NPF)is involved in the nitrate uptake,transport and distribution.Therefore,the genome-wide identification and expression analysis of the NRT/NPF gene family are important for the studying of nitrogen uptake and utilization in P.bournei.In the present study,the genome-wide NRT/NPF genes of P.bournei were identified using NRT protein sequences of Arabidopsis thaliana and Hidden Markov Model(HMM)(PF00854,PF07690,PF16974),and 63 PbNRT/NPF members were identified,which were unevenly distributed on 11 chromosomes.The results showed that the molecular weights of PbNRT/NPF were 22.8~135.6 kD,isoelectric point were 5.44~9.41,encoding 205 to 1235 aa.Most of the encoded proteins were basic proteins with 2~23 transmembrane domains,phylogenetic analysis revealed that 63 PbNRT/NPF genes could be divided into 3 subfamilies,of which 60 belonged to the NRT1/NPF subfamily,2 to the NRT2 subfamily subfamily and 1 to the NRT3 subfamily.Collinearity analysis revealed that there were 9 tandem duplication events and 10 segmental duplication events in the PbNRT/NPF family.The transcriptome data and qRT-PCR analysis showed that PbNRT/NPF members were differentially expressed among 5 tissues of P.bournei,for example,PbNPF6.3 was significantly higher expressed in the root bark than that of other tissues.Furthermore,PbNRT/NPF members showed differential expression patterns under different low nitrogen treatments,e.g.the expression of PbNRT2.4 and PbNRT2.5 significantly increased under low nitrogen treatment.Overall,this study provides a basis for studying the functions of the PbNRT/NPF genes in nitrogen absorption,transport and utilization in P.bournei,and provides a theoretical basis for breeding and selection of low-nitrogen tolerant varieties in P.b

关 键 词：闽楠 硝酸盐转运蛋白(NRT/NPF) 基因家族 低氮胁迫 表达分析 

分 类 号：S792.24[农业科学—林木遗传育种]