核桃铵态氮转运蛋白基因JrAMT2的功能分析  

Functional analysis of ammonium nitrogen transporter gene JrAMT2 in Juglans regia

作　　者：凡婷婷 张佳琦 刘会君 王凤敏 马宇航 吴宇伟 胡恒康[2] 黄有军[1,2] 李岩[2] 王克涛 黄坚钦[2] 张启香[1,2] FAN Tingting;ZHANG Jiaqi;LIU Huijun;WANG Fengmin;MA Yuhang;WU Yuwei;HU Hengkang;HUANG Youjun;LI Yan;WANG Ketao;HUANG Jianqin;ZHANG Qixiang(Zhejiang Provincial Key Laboratory of Forest Aromatic Plants-based Healthcare Functions,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;State Key Laboratory of Subtropical Silviculture,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China)

机构地区：[1]浙江农林大学浙江省森林芳香植物康养功能研究重点实验室,浙江杭州311300 [2]浙江农林大学省部共建亚热带森林培育国家重点实验室,浙江杭州311300

出　　处：《浙江农林大学学报》2024年第1期79-91,共13页Journal of Zhejiang A&F University

基　　金：浙江省重点研发计划项目(2021C02037);浙江省农业新品种选育重大科技专项(2021C02066-12);国家自然科学基金资助项目(31971672,32171815,32101557);浙江省自然科学基金资助项目(LY18C150002);浙江农林大学校级学生科研训练项目(S202210341173)。

摘　　要：【目的】研究高效利用氮素基因铵态氮转运蛋白基因JrAMT2,对核桃Juglansregia的品种改良、快速生长及产量形成有重要意义。【方法】以核桃JrAMT2过表达幼苗为实验材料,对JrAMT2基因进行生物信息学分析。通过基因表达量和表型测定对核桃JrAMT2过表达植株生长发育、氮素吸收、叶绿素质量分数、叶绿素荧光进行理化分析。【结果】JrAMT2基因在核桃JrAMT2过表达植株中稳定表达。与野生型相比,核桃JrAMT2过表达株系的株高、节间长、生物量等生长参数显著提高(P<0.05),核桃幼苗的株高、节间长增加,最高可增加68.2%和50.3%,植株地上部分、地下部分生物量显著增加,地上部分最高可增加56.26%(鲜质量)和56.26%(干质量),地下部分最高可增加344.38%(鲜质量)和354.33%(干质量);核桃JrAMT2过表达株系地下部分对铵态氮及硝态氮的吸收显著提高(P<0.05),最高可提高114.1%和70.3%,其中JrAMT2基因介导了铵态氮从地下部分到地上部分的运输,地上部分铵态氮质量分数显著增加(P<0.05),最高可增加59.1%;核桃JrAMT2过表达植株叶绿体表面积与单层细胞表面积比率、叶绿素质量分数显著增加(P<0.05),最高可增加22.94%和74.3%;对叶绿素荧光参数分析,核桃JrAMT2过表达植株叶片放氧复活体活性、量子产额、电子传递效率均显著提高(P<0.05)。【结论】核桃JrAMT2基因在核桃幼苗生长发育、氮素吸收和光合作用中均有积极显著调控的作用,为进一步研究核桃快速繁育提供一定的理论依据,且为筛选优良品种奠定一定基础。[Objective]The objective is to study the efficient utilization of nitrogen genes and the ammonium nitrogen transporter protein JrAMT2 gene,which is of great significance for variety improvement,rapid growth,and yield of Juglans regia.[Method]Using J.regia JrAMT2 overexpression(OE)seedlings as experimental materials,bioinformatics analysis was conducted on JrAMT2 gene.The growth and development,nitrogen absorption,chlorophyll content,and chlorophyll fluorescence of J.regia JrAMT2-OE plants were analyzed through gene expression level and phenotype determination.[Result]Fluorescence detection,polymerase chain reaction,and fluorescence quantitative PCR were performed on J.regia JrAMT2-OE plants to verify that the JrAMT2 gene is stably expressed in J.regia JrAMT2-OE plants.Compared with the wild type,the plant height,internode length,biomass and other growth parameters of JrAMT2-OE lines significantly increased,and the plant height and internode length of J.regia seedlings increased by 68.2%and 50.3%.The aboveground and underground biomass of JrAMT2-OE lines increased significantly,with a maximum increase of 56.26%(fresh weight)and 56.26%(dry weight)in the aboveground parts,and a maximum increase of 344.38%(fresh weight)and 354.33%(dry weight)in the underground parts.The uptake of ammonium nitrogen and nitrate nitrogen in the underground part of J.regia JrAMT2-OE significantly increased by 114.1%and 70.3%respectively.The JrAMT2 gene mediated the transport of ammonium nitrogen from the underground part to the aboveground part,and the content of ammonium nitrogen in the aboveground part increased significantly,up to 59.1%.The ratio of chloroplast surface area to monolayer cell surface area and chlorophyll content of JrAMT2-OE increased significantly,with the highest increase of 22.94%and 74.3%.The analysis of chlorophyll fluorescence parameters showed that the activity,quantum yield and electron transfer efficiency of the oxygen-reactive body in the leaves of J.regia JrAMT2-OE plants significantly improved.[Conclusion]T

关 键 词：核桃 JrAMT2 铵态氮转运蛋白 过表达 光合作用 

分 类 号：S722.3[农业科学—林木遗传育种] Q781[农业科学—林学]