荔枝愈伤组织继代及体胚发生过程中 结构与多胺含量的变化  

Changes in structure and polyamine metabolism of litchi callus during subculture and somatic embryo development

作　　者：王果[1] 刘耀婷 高兆银[1] 李焕苓[1] 王树军[1] 李芳 王家保[1] WANG Guo;LIU Yaoting;GAO Zhaoyin;LI Huanling;WANG Shujun;LI Fang;WANG Jiabao(Environments and Plant Protection Institute,Chinese Academy of Tropical Agriculture Sciences/Danzhou Scientific Observing and Experimental Station of Agro-Environment,Ministry of Agriculture and Rural Affairs,Danzhou 571737,Hainan,China;College of Tropical Crops,Hainan University,Haikou 570228,Hainan,China)

机构地区：[1]中国热带农业科学院环境与植物保护研究所·农业农村部儋州农业环境科学观测实验站,海南儋州571737 [2]海南大学热带作物学院,海口570228

出　　处：《果树学报》2021年第11期1911-1920,共10页Journal of Fruit Science

基　　金：国家重点研发计划(2019YFD1000900);中央级公益性科研院所基本科研业务费专项(1630042019026);国家现代农业产业技术体系(CARS-32-03)。

摘　　要：【目的】探究妃子笑荔枝愈伤组织继代及体胚发生过程中结构与多胺变化,为优化荔枝体胚发生技术体系提供理论依据与技术基础。【方法】采用石蜡切片技术、苏木精整体染色法、GC-MS法研究了胚性愈伤组织继代及体胚发生过程中的形态结构及多胺代谢变化。【结果】愈伤组织继代过程中,日增殖量前期增殖缓慢,后期快速增加,达到峰值后下降,最大日增殖量出现在继代后第15天。愈伤组织快速增殖阶段颗粒明显,细胞形态相对均匀,细胞质浓厚,染色深。在体胚发生培养基上约10 d时愈伤组织出现明显的厚壁细胞形成生殖隔离,分化原胚,在原胚发生及子叶胚时期有大量淀粉积累。腐胺(Put)和精胺(Spm)是荔枝愈伤组织中多胺(PAs)的主要组分,亚精胺(Spd)含量较低。Put在愈伤组织中维持在较高水平,Put与Spd含量在愈伤组织胚性较强(继代第6天及体胚发生第10天)时都达到最高,体胚发生时愈伤组织中的Spm含量远低于继代时含量。多胺氧化酶(PAO)及二胺氧化酶(DAO)活性与Put及PAs含量呈正相关。【结论】愈伤组织继代约第15天时日增殖量最大,体胚发生过程中约在第10天分化体胚。高的PAO活性与PAs含量有利于荔枝体胚高频发生。【Objective】Plant regeneration in vitro is the basis of transgenic breeding and rapid propagation of litchi(Litchi chinensis Sonn.).Components in the medium are the main factors that affect the efficiency of litchi regeneration in vitro.At present,research on medium components has focused mainly on cytokinin and auxin,and very few studies have been performed on polyamines.In this study,the anatomical structure,polyamine components,and activity of associated enzymes were investigated during the subculture and somatic embryo induction of Feizixiao embryonic callus so as to optimize in vitro regeneration of litchi.【Methods】Daily proliferation was measured during the subculture of embryonic callus.Structural characteristics of embryonic callus during different stages of subculture and somatic embryo induction were studied by paraffin section,and the polyamine content was tested by GC-MS.The activity of enzymes involved in polyamine metabolism was assayed using a colorimetric method.【Results】The daily proliferation of embryonic callus remained almost unchanged during the earlystage(0–6 d),after which it increased rapidly(7–15 d)to a peak at 15 d after subculture.After that,the proliferation decreased and then became relatively stable during the late stage(18–24 d).During the rapid proliferation stage,the granules of the embryonic callus were apparent.The paraffin sections of the callus showed that the cell divided vigorously,and the cell morphology was uniform with dense cytoplasm,and thus the sections were deeply stained.After inoculation on the somatic embryo induction medium,the callus was gradually dried and differentiated,and some early somatic embryos were visible at the early stage(0–10 d).Microscopic observation showed that there were apparent sclerenchyma cells in the callus,which resulted in obvious separation of the differentiated proembryos,and accumulation of many starch grains in the cytoplasm.After 15 d on the induction medium,the somatic embryo showed evident polarization.The cytopla

关 键 词：荔枝 愈伤组织 组织形态 多胺 多胺相关酶 

分 类 号：S667.1[农业科学—果树学]