八棱海棠高效再生体系的建立及潮霉素的抗性试验  

Establishment of Efficient Regeneration System and Study of Hygromycin Resistance Test of Malus robusta Rehd.

作　　者：张苓 彭日荷[2] 付晓燕[2] 高建杰[2] 王明清 姚泉洪[1,2] ZHANG Ling;PENG Rihe;FU Xiaoyan;GAO Jianjie;WANG Mingqing;YAO Quanhong(College of Horticulture,Nanjing Agricultural University,Nanjing,Jiangsu 210095;Biotechnology Research Institute,Shanghai Academy of Agricultural Sciences,Shanghai 201106)

机构地区：[1]南京农业大学园艺学院,江苏南京210095 [2]上海市农业科学院生物技术研究所,上海201106

出　　处：《北方园艺》2020年第15期82-88,共7页Northern Horticulture

基　　金：国家自然科学基金资助项目(41907097);上海市自然科学基金资助项目(18ZR1413100)。

摘　　要：以八棱海棠子叶为外植体,比较了不同浓度植物激素、不同激素组合、不同暗培养时间及不同浓度潮霉素对八棱海棠再生体系的影响,建立了八棱海棠高效再生体系,筛选并分析了潮霉素在八棱海棠遗传转化中的适宜浓度。结果表明:诱导子叶愈伤的最佳培养基为Sc+2.0 mg·L^-1 TDZ+0.2 mg·L^-1 NAA+0.5 mg·L^-1 GA3,芽再生的最佳培养基为Sc+2.0 mg·L^-1 TDZ+0.2 mg·L^-1 NAA,暗培养最佳时间为10 d,生根最佳培养基为1/2MS+0.25 mg·L^-1 IBA。八棱海棠在整个再生过程中均对潮霉素敏感,在潮霉素浓度为2.0 mg·L^-1的愈伤诱导培养基上,外植体的愈伤诱导率仅为23.33%,分化率仅为9.88%;当潮霉素浓度为4.0 mg·L^-1时,愈伤诱导率下降为13.00%,而且愈伤不分化,随着培养时间延长,愈伤组织变得更加致密,之后变褐死亡。在诱导生根培养基上添加3.0 mg·L^-1潮霉素,生根率仅为3.52%,大部分外植体再生芽底部只会诱导出愈伤组织,没有根原基形成。八棱海棠子叶愈伤诱导及分化的潮霉素筛选临界浓度为2.0~4.0 mg·L^-1,生根时潮霉素筛选浓度为2.0~3.0 mg·L^-1。In order to establish efficient regeneration system and study the effect of hygromycin concentration on cotyledon of Malus robusta Rehd.differentiation and rooting,different plant hormone concentrations and culture methods were used.The results showed that the best medium to induce callus was Sc+2.0 mg·L^-1 TDZ+0.2 mg·L^-1NAA+0.5 mg·L^-1GA3.The best medium for adventitious bud regeneration was Sc+2.0 mg·L^-1 TDZ+0.2 mg·L^-1NAA.The optimum time for dark culture was 10 days.The optimal rooting medium was 1/2 MS+0.25 mg·L^-1 IBA.Malus robusta Rehd.was found to be sensitive to hygromycin by the growth of cotyledon in medium with different concentration of hygromycin.In the medium with hygromycin concentration of 2.0 mg·L^-1,the callus induction rate of explants was only 23.33%,and the budding rate was even lower at 9.88%.When the concentration of hygromycin was increased to 4.0 mg·L^-1,the callus induction rate of explants was only 13.00%,and allus was produced,but the callus did not differentiate.With the extension of culture time,the callus became denser,then browned and died.During rooting induction,the rooting rate was 3.52%in medium with hygromycin 3.0 mg·L^-1,and the explants showed callus at the bottom of the stem,but no root primordium was formed.Therefore,the critical screening concentration of Malus robusta Rehd.differentiation element was 2.0-4.0 mg·L^-1,and the screening concentration of rooting hygromycin was 2.0-3.0 mg·L^-1.

关 键 词：八棱海棠 再生体系 潮霉素 临界浓度 

分 类 号：S661[农业科学—果树学]