柳枝稷人工穗芽块遗传转化体系的初步建立  

Preliminary Establishment of Genetic TransformationSystem for Artificial Spike Bud Blocks in Switchgrass

作　　者：柴乖强 韦明月 封文佳 李琳 段义忠[1,2] 范希峰 CHAI Guai-qiang;WEI Ming-yue;FENG Wen-jia;LI Lin;DUAN Yi-zhong;FAN Xi-feng(College of Life Science,Yulin University,Yulin 719000,China;Shaanxi Key Laboratory of Ecological Restoration in Shaanbei Mining Area,Yulin 719000,China;Beijing Academy of Agriculture and Forestry Sciences Beijing Research﹠Development Center for Grass and Environment/Key Laboratory of Urban Agriculture(North China),Ministry of Agriculture and Rural Affairs,Beijing 100097,China)

机构地区：[1]榆林学院生命科学学院,陕西榆林719000 [2]陕西省陕北矿区生态修复重点实验室,陕西榆林719000 [3]北京市农林科学院草业与环境研究发展中心/农业农村部都市农业(北方)重点实验室,北京100097

出　　处：《中国草地学报》2022年第7期13-22,共10页Chinese Journal of Grassland

基　　金：国家自然科学基金项目(32160464);陕西省教育厅科研计划项目(21JS042);榆林学院博士科研启动基金项目(18GK11)。

摘　　要：以柳枝稷Alamo和Cave in rock两个基因型的471块人工穗芽块为外植体,采用农杆菌介导法转化糜子叶片表皮蜡质合成基因PmWX2,经过共培养、抗性穗芽块筛选和生根,共获得了30株转化植株,PCR检测分析表明目的基因PmWX2已整合到柳枝稷基因组中,其最高转化效率为1.23%。对柳枝稷转基因植株苗期叶片的蜡质含量分析结果表明,4个转PmWX2基因株系中,Line 2和Line 8的蜡质总量分别是对照的1.7倍和1.5倍,显著高于对照(P<0.05);在转基因植株的蜡质成分中,变化最明显的是β-二酮和OH-β-二酮,株系Line 2的β-二酮和OH-β-二酮分别比对照增加了134.2%和184.9%,说明糜子PmWX2基因在株系Line 2中已经表达。表皮蜡质晶体结构分析显示,转基因株系Line 2叶片反面棒状蜡质晶体较野生型显著增多,说明蜡质合成基因PmWX2促进了柳枝稷叶片表皮棒状蜡质晶体的合成及分泌。人工穗芽块遗传转化体系的建立为柳枝稷的基因功能分析提供了方法支撑。In order to explore the genetic transformation mechanism of switchgrass,471 artificial panicle bud tissues of switchgrass“Alamo”and“Cave in Rock”were used as explants,and the agrobacterium-mediated transformation method was used to transform the epidermal wax synthesis gene PmWX2 of millet leaves.After co-culturing,screening of resistant panicle bud and rooting,a total of 30 transformed plants were obtained.PCR(Polymerase chain reaction)analysis indicated that the target gene PmWX2 had been integrated into the switchgrass genome,and the highest transformation efficiency was 1.23%.The analysis of wax content in leaves of switchgrass transgenic plants at seedling stage showed that the total wax content of Line 2 and Line 8 in the four PmWX2 transgenic lines was 1.7 and 1.5 times higher than that of control,respectively(P<0.05).Among the waxy components of transgenic plants,β-diketone and OH-β-diketone change was the most obvious,and Line 2β-diketone and OH-β-diketone increased by 134.2%and 184.9%,respectively,compared with the control,indicating that the PmWX2 gene of millet has been expressed in Line 2.The SEM(Scanning electron microscope)analysis of epidermal waxy crystal structure showed that the number of rod-shaped waxy crystals on the leaf abaxial surface of transgenic Line 2 was significantly more than that of wild type,indicating that the wax synthesis gene PmWX2 promoted the synthesis and secretion of rod-shaped waxy crystals on the epidermis of switchgrass leaves.These results provide a feasible method for gene function analysis of energy crops such as switchgrass.

关 键 词：柳枝稷 人工穗芽 遗传转化 表皮蜡质 

分 类 号：S54[农业科学—作物学]