基于叶绿素合成关键基因构建马蹄莲佛焰苞离体VIGS技术体系  

Establishing of an In Vitro Spathe Virus Induced Gene Silencing(VIGS)System for Calla Lily(Zantedeschia)Based on Chlorophyll Synthesis Genes

作　　者：刘红艳 杨拓 王壹 孙轩 王贤[2] 刘振林[1] 张国君[1] 卫尊征[2] LIU Hongyan;YANG Tuo;WANG Yi;SUN Xuan;WANG Xian;LIU Zhenlin;ZHANG Guojun;WEI Zunzheng(College of Horticultural Science&Technology/Hebei Key Laboratory of Horticultural Germplasm Excavation and Innovative Utilization/Hebei Higher Institute Application Technology Research and Development Center of Horticultural Plant Biological Breeding,Hebei Normal University of Science&Technology,Qinhuangdao,Hebei 066000;Institute of Grassland,Flowers and Ecology,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097;College of Horticulture,China Agricultural University,Beijing 100193)

机构地区：[1]河北科技师范学院园艺科技学院/河北省特色园艺种质挖掘与创新利用重点实验室/河北省高校特色园艺植物生物育种应用技术研发中心,河北秦皇岛066000 [2]北京市农林科学院草业花卉与景观生态研究所,北京100097 [3]中国农业大学园艺学院,北京100193

出　　处：《核农学报》2023年第7期1289-1298,共10页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金面上项目(32071812);北京市农林科学院创新能力建设项目(KJCX20200111);河北省级专业学位研究生培养实践基地(202018);“十二五”农村领域国家科技计划课题(2012AA102002-5)。

摘　　要：返青是天南星科观赏植物在发育过程中发生的一种典型的功能叶绿体重新分化形成的生理现象,其中马蹄莲佛焰苞常出现这一现象,但其潜在的调控机制并不明确。为了探究马蹄莲返青的调控机制,提高彩色马蹄莲的观赏品质,本研究以黄色品种Florex Gold为材料,以返青过程中两个叶绿素合成关键基因ZhGluTR和ZhChlH为研究对象,建立病毒诱导的基因沉默(VIGS)体系,对马蹄莲进行基因功能验证。通过离体佛焰苞孔片试验并对取样部位和侵染时长进行综合分析,结果显示,中部真空抽吸5 min的试验组返青现象出现最迟,VIGS沉默效果最好。在上述沉默背景下,与对照组相比,ZhGluTR和ZhChlH沉默孔片的返青现象均出现得更迟,色调角值上升更缓慢,并且叶绿素含量更低,类胡萝卜素含量更高。综合上述结果表明,本研究建立的VIGS体系可分别有效沉默佛焰苞中的叶绿素合成关键基因ZhGluTR和ZhChlH。研究结果有助于进一步探究马蹄莲佛焰苞返青现象的分子调控机制。Regreening is a typical physiological phenomenon of functional chloroplast re-differentiation during the development in Araceae ornamental plants,this phenomenon often occurs in the spathe of calla lily(Zantedeschia hybrida),but the underlyingly regulatory mechanism remains unclear.In this study,in order to explore the regreening mechanism and improve the ornamental quality of calla lilies,a virus induced gene silencing(VIGS)system was established to verify the gene function of two key genes in chlorophyll biosynthesis during regreening,ZhGluTR and ZhChlH using the yellow variety Florex Gold of calla lily as the material.An integrated evaluation of sample locations and duration of infiltration in isolated spathe discs indicated that,the test group with 5 min of vacuum extraction in the middle spathe showed the latest regreening and the best silencing effect.Compared with the control,the silenced spathe disc of ZhGluTR and ZhChlH exhibited delayed regreening,the hue angle value increased slower.In addition,chlorophyll was less and carotenoids were more.The above results showed that VIGS system in this study can effectively silence the key chlorophyll synthesis genes ZhGluTR and ZhChlH in the spathe.The results of this study laid a foundation for revealing the molecular regulation mechanism of spathe regreening in calla lily in the future.

关 键 词：马蹄莲 佛焰苞孔片 病毒诱导的基因沉默(VIGS) 基因功能 

分 类 号：S682.264[农业科学—观赏园艺]