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矮牵牛Ph4CL13基因的功能鉴定及分析OA北大核心CSTPCD

Functional Identification and Analysis of Ph4CL13 Gene in Petunia(Petunia hybrida)

中文摘要英文摘要

花青苷是影响植物花瓣着色的主要色素.根据课题组前期矮牵牛(Petunia hybrida)转录组数据筛选出在花蕾着色期高表达的4香豆酰辅酶A连接酶13(4-coumarate CoA ligase 13,Ph4CL13)基因,推测其与花青苷合成有关.为研究Ph4CL13在花青苷合成中的功能,本研究以矮牵牛'紫罗兰'作为材料,通过矮牵牛基因组筛选并克隆得到长度为1 653 bp的Ph4CL13(Peaxi162Scf00089g00045.1)cDNA片段.系统发育树分析显示,Ph4CL13与草莓(Fragaria×ananassa)Fa4CL-1和Fa4CL-2亲缘关系较近.qPCR分析Ph4CL13在矮牵牛花开放不同阶段的表达模式发现,Ph4CL13在矮牵牛花蕾着色期高表达,推测Ph4CL13可能参与花青苷合成从而影响矮牵牛花色.通过病毒诱导的基因沉默(virus induced gene silencing,VIGS)技术沉默Ph4CL13,结果显示,Ph4CL13沉默后矮牵牛花色变白,花青苷水平较对照显著降低;qPCR 分析结果显示,沉默 Ph4CL13 后使其下游多个花青苷合成相关基因查尔酮合成酶(chalconesynthase,CHS)、查尔酮异构酶(chalcone isomerase,CHI)、黄烷酮-3-羟化酶(flavanone-3-hydroxylase,F3H)、二氢黄酮醇还原酶(dihydroflavonol 4-reductase,DFR)、黄酮醇合成酶(flavonoid synthase,FLS)的转录被抑制,表明Ph4CL13在矮牵牛花蕾着色期的花青苷合成阶段发挥重要作用,且编码花青苷合成酶的基因间可能存在复杂的反馈机制.本研究初步揭示了Ph4CL13在花青苷合成中的作用,为后续深入解析其在花色形成中的分子机制提供参考.

The main pigment responsible for the coloration of plant petals is anthocyanin.Based on previous transcriptome data from petunias,the highly expressed 4-coumarate CoA ligase 13(Ph4CL13)gene was identified during the bud coloring stage,suggesting its involvement in anthocyanin synthesis.Anthocyanin primarily determines petal color in plants.To investigate the role of Ph4CL13 in anthocyanin synthesis,this study used Petunia hybrida'Ultra'as the material.The 1 653 bp cDNA fragment of Ph4CL13(Peaxi162Scf00089g00045.1)was isolated from the petunia genome.Phylogenetic analysis revealed a close relationship between Ph4CL13 and Fa4CL-1 and Fa4CL-2.qPCR analysis revealed that Ph4CL13 was highly expressed during the coloring stages,suggested its involvement in anthocyanin synthesis and impact on petunia flower coloration.Silencing Ph4CL13 via virus-induced gene silencing(VIGS),the results showed that the corollas of petunia turned white,and the level of anthocyanin significantly decreased in Ph4CL13-silenced plant compared to control.qPCR analyses demonstrated that Ph4CL13 silencing transcriptionally repressed several anthocyanin biosynthesis genes,including chalconesynthase(CHS),chalcone isomerase(CHI),flavanone-3-hydroxylase(F3H),dihydroflavonol 4-reductase(DFR),flavonoid synthase(FLS),suggested that Ph4CL13 played a critical role in the anthocyanin synthesis stage of flower coloring process,that might be a potential complex feedback mechanism among these genes related to anthocyanin synthesis.This study preliminarily revealed the important role of Ph4CL13 in anthocyanin synthesis,providing a reference for further exploring its molecular mechanisms in flower color development.

缪云锋;胡旭浩;林彬茹;钟诗蔚

浙江农林大学风景园林与建筑学院/浙江省园林植物种质创新与利用重点实验室,杭州 311300

园艺学与植物营养学

矮牵牛花青素苷4香豆酰辅酶A连接酶(4CL)病毒介导的基因沉默(VIGS)

Petunia hybridaAnthocyanins4-coumarate CoA ligase(4CL)Virus induced gene silencing(VIGS)

《农业生物技术学报》 2024 (006)

1274-1283 / 10

浙江农林大学科研启动基金(203402010601);浙江农林大学大学生创新训练项目(2021KX0070)

10.3969/j.issn.1674-7968.2024.06.004

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