万寿菊雄性不育两用系遗传转化体系的建立  

Establishment of Genetic Transformation System for Marigold Male Sterile Two-type Line

作　　者：陈利文 唐楠[1,2] 张五华 唐道城[1,2] 尹计成 张金艳 陈梦茹 冯依玲 姚文周 杨红峡 Chen Liwen;Tang Nan;Zhang Wuhua;Tang Daocheng;Yin Jicheng;Zhang Jinyan;Chen Mengru;Feng Yiling;Yao Wenzhou;Yang Hongxia(Plateau Flower Research Center,Qinghai University,Xining,810016;Qinghai Provincial Key Laboratory of Ornamental Plants and Ornamental Horticulture,Qinghai University,Xining,810016;Qinghai Tongyi Ecological Management Co.,Ltd.,Xining,810000)

机构地区：[1]青海大学高原花卉研究中心,西宁810016 [2]青海大学,青海省园林植物与观赏园艺重点实验室,西宁810016 [3]青海通艺生态治理有限公司,西宁810000

出　　处：《分子植物育种》2023年第19期6398-6405,共8页Molecular Plant Breeding

基　　金：青海省中央引导地方科技发展资金项目(2021ZY025)。

摘　　要：为了筛选适合万寿菊不定芽诱导的最佳外植体及激素浓度组合,并探讨该组织部位对卡那霉素、潮霉素的抗性临界点以及头孢霉素和利福平对LBA4404的抑菌性效果,构建万寿菊遗传转化体系。本研究以三种激素为组合,以万寿菊雄性不育两用系2-2的子叶、下胚轴、叶片为材料筛选出适合万寿菊的再生体系的激素配比及最佳的组织部位;以万寿菊子叶为材料筛选其对卡那霉素、利福平、头孢霉素、潮霉素四种抗生素抗性临界点;筛选利福平、头孢霉素对农杆菌LBA4404的抑菌浓度。结果表明:1 mg/L 6-BA+0.1 mg/L IAA+30 g蔗糖的MS培养基可以作为万寿菊子叶的最佳诱导激素浓度组合,MS5和MS9可以在万寿菊叶片中诱导出不定芽,但诱导率极低,下胚轴在48个激素组合无不定芽的产生;利福平和头孢霉素两种抗生素浓度在200 mg/L时能对LBA4404农杆菌的生长起到有效的抑制,并且在该浓度下对万寿菊子叶不定芽的产生起到一定的促进作用;子叶对于潮霉素、卡那霉素的抗性浓度分别为30、100 mg/L。利用建立的万寿菊再生体系及抗生素浓度体系可以为农杆菌介导万寿菊遗传转化提供理论基础。In order to screen the best combination of explants and hormone concentration suitable for induction of adventitious buds of marigold,and to explore the critical point of resistance of this tissue site to kanamycin and hygromycin and the inhibition of cephalosporin and rifampicin on LBA4404 bacterial effect,the genetic transformation system of marigold was constructed.In this study,three hormones were used as a combination,and the cotyledons,hypocotyls,and leaves of the marigold male sterility line 2-2 were used as materials to screen out the hormone ratio and the best tissue position suitable for the regeneration system of marigold.Use marigold cotyledons as materials to screen the critical points of resistance to kanamycin,rifampicin,cephalosporin and hygromycin.Screening the inhibitory concentration of rifampicin and cephalosporin against A grobacterium LBA4404.The results showed that the MS medium of 1 mg/L 6-BA+0.1 mg/L IAA+30 g sucrose was the best combination of inducing hormone concentration in marigold cotyledons,and MS5 and MS9 could induce adventitious buds in marigold leaves,but the induction rate was extremely low.Hypocotyls did not produce adventitious buds in 48 hormone combinations.Rifampicin and cephalosporin at 200 mg/L could effectively inhibit the growth of A grobacterium LBA4404,and at this concentration,the adventitious bud production of marigold cotyledons was promoted to a certain extent.The resistance concentration of cotyledons to hygromycin and kanamycin were 30 and 100 mg/L,respectively.The established marigold regeneration system and antibiotic concentration system can provide a theoretical basis for A grobacterium-mediated genetic transformation of marigold.

关 键 词：万寿菊 抗生素 LBA4404农杆菌 再生体系 遗传转化体系 

分 类 号：S681.9[农业科学—观赏园艺]