基于VIGS技术的无花果FcMYB114基因功能研究  

Function of FcMYB114 gene in fig(Ficuscarica L.) by virus⁃induced gene silencing

作　　者：陈紫玉 高文科 李明海 何弟南 龙艺丹 黄蕾 王梓然[1] CHEN Zi-yu;GAO Wen-ke;LI Ming-hai;HE Di-nan;LONG Yi-dan;HUANG Lei;WANG Zi-ran(School of Landscape Architecture and Horticulture,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学园林园艺学院,昆明650201

出　　处：《西南农业学报》2023年第3期623-628,共6页Southwest China Journal of Agricultural Sciences

基　　金：云南省基础研究专项-青年项目(202101AU070094);云南省教育厅科学研究基金项目(2020J0247);云南农业大学优秀本科毕业设计(论文)培育专项(2021YPX003)。

摘　　要：【目的】无花果紫皮品种相比于青皮品种,具有外观品质好、保健价值高、商品价值高等优势,是目前无花果育种的主要方向。对调控无花果果皮颜色的相关基因进行功能性研究对于选育不同皮色的无花果品种具有重要意义。【方法】以‘紫宝’无花果为试材,利用同源克隆技术对‘紫宝’无花果果皮类黄酮生物合成调控基因FcMYB114的保守域进行克隆,通过VIGS(Virus induced gene silencing)技术将FcMYB114基因进行沉默,并对其功能进行研究。【结果】基因定量表达分析显示,FcMYB114基因在‘紫宝’幼果期果皮(PY)中表达量最少,在成熟期果皮(PM)中表达量最高,通过遮光处理后FcMYB114基因的表达量显著降低;系统进化树表明,无花果FcMYB114(c42269_g1)与红皮梨PyMYB114的亲缘关系最近,序列相似性高达83.12%,与拟南芥中AtMYB114、碧桃中PpMYB9亲缘关系较近;多重序列比对分析结果显示,FcMYB114在序列N端具有高度同源的R2-R3 DNA结合结构域,与大多数MYB转录因子的结构相似;经VIGS沉默处理的无花果果实与对照相比,在果皮色泽上存在明显差异,花色苷含量约为对照的38.5%;RT-qPCR分析表明类黄酮生物合成代谢途径相关结构基因在FcMYB114基因沉默处理后其表达量发生改变,其中FcDFR、FcANS和FcUFGT表达量呈显著下调趋势。【结论】在pTRV2-FcMYB114介导的无花果果皮中,花青素含量降低,类黄酮生物合成代谢途径结构基因表达量显著下调,表明FcMYB114基因可能正向调控无花果果皮花青素合成,为无花果类黄酮生物合成途径中相关转录因子的分子调控机理解析提供了新的见解,为创制不同色泽的无花果品种奠定理论基础。【Objective】Purple⁃skinned varieties of figs have the advantages of better appearance quality,higher health value,and higher com⁃mercial value than green⁃skinned varieties,making them the main direction of the fig breeding at present.The functional study on the genes related to the regulation of fig peel color is of great significance for the selection of fig varieties with different peel colors.【Method】Using the‘Zibao’fig as the test material,the conservative domain of theFcMYB114fruit peel flavonoid biosynthesis regulatory gene of the‘Zibao’fig was cloned by using homologous cloning technology.The geneFcMYB114was silenced by VIGS(Virus⁃induced gene silencing)technol⁃ogy,and its function was studied.【Result】Quantitative gene expression analysis revealed thatFcMYB114was expressed at the lowest level in the young fruit peel(PY)of‘Zibao’and at the highest level in the mature fruit peel(PM),and its expression was significantly reduced af⁃ter light deprivation treatment.The phylogenetic tree showed thatFcMYB114(c42269_g1)of fig had the closest relationship withPyMYB114 of red⁃skinned pear,with a sequence similarity of 83.12%,and was also closely related toAtMYB114inArabidopsisandPpMYB9inPrunus persica.The multiple sequence alignment analysis results showed thatFcMYB114had a highly homologous R2⁃R3 DNA binding domain at the N⁃terminal,which was similar to the structure of most MYB transcription factors.The fig fruits silenced by VIGS showed obvious differ⁃ences in fruit peel color compared with the control,and the anthocyanin content was about 38.5%of the control.The RT⁃qPCR analysis showed that the expression levels of structural genes related to flavonoid biosynthesis metabolic pathway changed afterFcMYB114gene silen⁃cing,among whichFcDFR,FcANSandFcUFGTshowed a significant downward trend.【Conclusion】In the fig pericarp mediated by pTRV2⁃FcMYB114,the anthocyanin content decreased and the expression of structural genes in the flavonoid biosynthesis metabolic pathway

关 键 词：无花果 VIGS FcMYB114 类黄酮 花色苷 

分 类 号：S663[农业科学—果树学]