摘要
【目的】探究黄瓜CsMADS08基因及其下游调控基因的功能,为黄瓜新品种选育提供参考。【方法】以拟南芥agl8纯合突变体为材料,农杆菌介导法导入黄瓜CsMADS08基因,经筛选获得转CsMADS08抗性植株,比较agl8突变体植株、转CsMADS08植株及野生型拟南芥植株的表型变化,观察不同类型植株茎生叶的显微结构,并用实时荧光定量PCR方法分析CsMADS08及其下游调控基因SHP1、SHP2、ALC、IND在根、茎、莲座叶、茎生叶、花、果实中的表达情况。【结果】agl8突变体植株、转CsMADS08植株及野生型拟南芥根长、株高、莲座叶的数目及形状差异不明显;与野生型植株相比,agl8突变体植株抽薹及开花时间延迟,CsMADS08基因在agl8突变体植株花中的表达量无显著变化;而转CsMADS08植株抽薹及开花时间较agl8突变体植株提早4~6 d,且CsMADS08基因在花中表达量显著增加。野生型、agl8突变体及转CsMADS08拟南芥植株茎生叶数量、形状及侧枝数、果实均有明显差异,其中转CsMADS08植株叶片小而密集,形状接近圆形,果荚长且饱满,无提前开裂现象。茎生叶显微观察结果显示,与野生型植株相比,转CsMADS08植株叶片细胞变大,排列整齐,维管束细胞增加,叶片变厚。实时荧光定量PCR检测结果显示,CsMADS08基因在转CsMADS08植株各组织中均有表达,且在茎生叶中表达量最高;与agl8突变体植株相比,SHP1、SHP2、ALC、IND基因只在转CsMADS08植株花中上调表达,而在其他组织中均下调表达,且在茎生叶中降幅最大。【结论】CsMADS08基因具有促进开花、增加侧枝、调控叶片形态及果实发育等的功能。
【Objective】This study investigated the functions of CsMADS08 and its downstream regulatory genes to provide basis for cultivating new cucumber varieties.【Method】Cucumber CsMADS08 gene was introduced to Arabisopsis agl8 homozygous mutant by Agrobacterium-mediated method.The CsMADS08 transgenic resistant plants were obtained through screening.The phenotypic changes of the Arabidopsis agl8 mutant plants,transgenic plants and wild-type plants were compared.The structure of cauline leaves of different plants was observed by microstructures.The expressions of CsMADS08 and downstream regulatory genes SHP1,SHP2,ALC and IND in roots,stems,rosette leaves,stem leaves,flowers and fruits were analyzed by real-time fluorescence quantitative PCR.【Result】There were no significant diff-erences in root length,plant height,number and shape of rosette leaves between agl8 mutant plants,CsMADS08 transgenic plants and wild-type Arabidopsis.Compared with wild-type plants,the bolting and flowering time of agl8 mutant plants was delayed.There was no remarkable change in the expression of CsMADS08 gene in flowers of the agl8 mutant plants.The bolting and flowering time of CsMADS08 transgenic plants was 4-6 days earlier than that of the agl8 mutant plants and the expression of CsMADS08 gene in the flower of transgenic plants was increased.There were significant differences in number of branches,fruits,number and shape of stem leaves between three types of plants.The CsMADS08 transgenic plants had small and dense leaves,an approximate circular shape,as well as long and full fruit pods without premature cracking.Microscopic observation of stem leaves showed that,the leaf cells of CsMADS08 transgenic plants enlarged and arranged orderly compared with wild-type plants.The vascular bundle cells increased,and the leaves became thicker.Real-time fluorescent quantitative PCR results showed that CsMADS08 gene expressed in all tissues of the transgenic plants with the highest expression in stem leaves.In comparison with agl8 mutant plants,SHP1,SHP2,ALC and IND genes were up-regulated in flowers and down-regulated in other tissues of the CsMADS08 transgenic plants with the largest decrease in stem leaves.【Conclusion】CsMADS08 gene had the functions of promoting flowering,increasing lateral branches,and regulating leaf morphology and fruit development.
作者
许双双
安玉兰
温梦梅
胡克玲
杨咏
甘德芳
XU Shuangshuang;AN Yulan;WEN Mengmei;HU Keling;YANG Yong;GAN Defang(College of Horticulture,Anhui Agricultural University,Hefei,Anhui 230036,China;Jiangtang Agricultural Comprehensive Service Center,Taihu,Anhui 246417,China)
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2022年第2期115-124,共10页
Journal of Northwest A&F University(Natural Science Edition)
基金
安徽省自然科学基金项目(2008085MC80)
安徽省重点研究与开发计划项目(202004f06020047)
安徽省自然科学研究重点项目(KJ2019A0188,KJ2018A0154)
安徽农业大学2021年度研究生创新基金项目(2021ysj-8)。
