大麦旗叶距对小孢子发育时期、愈伤组织诱导以及绿苗再生的影响  

Effects of the flag leaf space on microspore developmental stage,callus induction and green plant regeneration in barley

作　　者：高润红[1] 郭桂梅[1] 何婷[1] 杜志钊 任金宝 刘成洪[1] 陆瑞菊[1] GAO Runhong;GUO Guimei;HE Ting;DU Zhizhao;REN Jinbao;LIU Chenghong;LU Ruiju(Biotech Research Institute/Shanghai Key Laboratory of Agricultural Genetics and Breeding,Shanghai Academy of Agricultural Sciences,Shanghai 201106,China;Shanghai Municipal Agricultural Science and Technology Service Centre,Shanghai 200335,China;Suzhou Polytechnic Institute of Agriculture,Suzhou 215008,China)

机构地区：[1]上海市农业科学院生物技术研究所/上海市农业遗传育种重点实验室,上海201106 [2]上海市农业科技服务中心,上海200335 [3]苏州农业职业技术学院,江苏苏州215008

出　　处：《南京农业大学学报》2021年第1期36-41,共6页Journal of Nanjing Agricultural University

基　　金：上海市科技兴农项目(2019-02-08-00-08-F01109);国家重点研发计划项目(2018YFD1000702-5);国家现代农业产业技术体系建设专项资金(CARS-05-01A-02)。

摘　　要：[目的]为提高大麦游离小孢子培养效率,在幼穗取材时建立更为精细的形态学指标,以获得更多处于适宜时期的小孢子进行培养。[方法]以大麦(Hordeum vulgare L.)品种‘花30’为材料,将同一天取材的幼穗根据旗叶与倒2叶的叶耳间距离(旗叶距)分为6组,观察每组材料低温处理前、后小孢子发育时期的变化,研究不同旗叶距对小孢子愈伤组织诱导及绿苗再生的影响,鉴定不同分组材料来源的小孢子再生植株的倍性。[结果]取材当天的幼穗不论旗叶距大小,所有游离的小孢子均处于单核期;而低温处理17 d后,处于单核时期的小孢子比例随旗叶距的增大而显著降低。随旗叶距增大,小孢子愈伤产量提高但差异不显著;而绿苗分化数量却显著降低。按旗叶距分组获得的小孢子再生植株不同倍性比例分布相近,二倍体占70%左右,单倍体和四倍体为30%左右。对于‘花30’而言,0.5~2.0 cm旗叶距的小孢子再生绿苗数量最高。[结论]旗叶距可作为大麦小孢子培养中幼穗取材的一个形态学指标,适宜的旗叶距可确保低温处理后获得更多单核时期游离小孢子和更高的再生绿苗数量。[Objectives]In order to improve the efficiency of isolated microspore culture for barley,more precise morphological indicator on sampling of young spikes is required to obtain more microspores at the suitable period for culture.[Methods]The sampled young spikes from barley variety‘Hua 30’were classified into six groups according to the distance between the ligules of the flag leaf and the penultimate leaf(space of flag leaf).The changes of isolated microspores developmental stage before and after low temperature treatment were observed in each group.Meanwhile,the effects of different group on production of callus induction and green plantlet regeneration were studied.In addition,the ploidy levels of regenerated plants originated from different groups were determined.[Results]All microspores isolated on the same sampling day were at mononuclear stage,no matter what flag leaf distance was.However,the proportion of mononuclear microspores decreased with the increase of flag leaf distance after 17 days of low-temperature treatment.Although the yields of induced callus did not improve significantly with the increase of the flag leaf space,the numbers of green plantlets decreased significantly.The proportion of ploidy levels for regenerated green plants via isolated microspore culture was similar during the groups of sampling spikes,diploids accounting for about 70%while haploids and tetraploids accounting for about 30%.For barley variety‘Hua 30’,the space of flag leaf between 0.5 and 2.0 cm was recommended for sampling of young spikes,which resulted in the highest production of regenerated green plants from isolated microspore culture.[Conclusions]The space of flag leaf could serve as a morphological indicator for sampling of young spikes,and the appropriate distance ensures more mononuclear microspores isolated after long term cold pretreatment and higher production of green plants for microspore culture in barley.

关 键 词：大麦 旗叶距 小孢子 愈伤诱导 再生能力 

分 类 号：S512.3[农业科学—作物学]