红松EC非胚性化及体胚分化启动过程的形态与生理特征  

Morphological and physiological characteristics of embryogenic callus of Pinus koraiensis during non embryogenization and initial somatic embryo differentiation

作　　者：钟玥 范震宇[1] 贯春雨 曹业萍 李青晨 梁艳[1] ZHONG Yue;FAN Zhenyu;GUAN Chunyu;CAO Yeping;LI Qingchen;LIANG Yan(College of Life Sciences and Agriculture and Forestry,Qiqihar University,Qiqihar 161006,Heilongjiang,China)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006

出　　处：《中南林业科技大学学报》2024年第11期49-57,共9页Journal of Central South University of Forestry & Technology

基　　金：国家自然科学基金项目(31800515);黑龙江省自然科学基金项目(LH2022C102);黑龙江省教育厅基本业务专项(145109137);大学生创新创业训练计划项目(x202310232028)。

摘　　要：【目的】研究长期继代的红松胚性愈伤组织(embryogenic callus,EC)非胚性化及体胚(somatic embryogenesis,SE)分化启动的生理特性,探讨不同胚性的愈伤组织形态特征与生理特性对应关系,为突破红松体胚发生中的EC胚性难以保持的技术瓶颈提供参考。【方法】本研究以红松长期继代获得的不同类型愈伤组织(分别为继代2个月和继代6个月的白色、黄色、水渍型、纤维化、褐化、含早期球形胚的愈伤组织)为材料,比较分析不同类型愈伤组织的形态特征、生理指标、体胚分化能力的差异。【结果】继代保持2~6个月获得的6种类型红松愈伤组织在颜色、质地等外部形态与组织细胞学特征,以及增殖速度与体胚分化能力方面差异明显,在继代2个月的白色高胚性愈伤组织(C1)中观察到明显的原胚团(PEMs)结构,并且这种类型愈伤组织能分化出球形胚,在其他5种类型的愈伤组织(C2-C6)中观察到PEMs结构的解体,以及大量形状不规则的细胞和降解的细胞,未见有球形胚的分化。高胚性的愈伤组织中可溶性糖及淀粉含量最高;高活性POD出现在高胚性的愈伤组织和褐化的愈伤组织中;SOD活性在高胚性的愈伤组织和有球形胚分化的愈伤组织中较低,而在增殖速度快的黄色愈伤中SOD活性较高;有球形胚分化的愈伤组织和低胚性的白色愈伤组织CAT活性最高;水渍型和纤维化的愈伤组织SOD、POD、CAT活性均较低。【结论】长期继代的红松EC保持胚性或失去胚性以及体胚分化启动过程受细胞内保护酶和贮藏类物质协同调控,高含量的可溶性糖和淀粉利于红松EC维持胚性,SOD、POD、CAT这3种关键酶在不同颜色质地的红松愈伤组织形成中的作用存在差异。【Objective】We investigated the physiological characteristics of non embryogenization and somatic embryogenesis(SE)differentiation initiation in long-term subcultured embryogenic callus(EC)of Pinus koraiensis,explored the correspondence between morphological and physiological characteristics of the different embryogenic potential calli,which can provide a reference for overcoming the technical bottleneck that the embryogenetic potential of EC is difficult to maintain in SE of P.koraiensis.【Method】In this study,different types of calli(white calli subcultured for 2 months and 6 months,yellow calli,water-stained calli,fibrotic calli,browned calli,and calli containing early spherical embryos,respectively)obtained by long-term subgeneration of.koraiensis were used as materials,and the differences in morphological characteristics,physiological indexes,and somatic embryo differentiation ability of different types of calli were compared and analyzed.【Result】The six types calli of Korean pine obtained by subcultured for 2-6 months had significant differences in external morphological and histocytological characteristics such as color and texture,as well as proliferation rate and somatic embryo differentiation ability. In the 2-month-old white calli with high embryogenic potential (C1), a distinct proembryogenic masses (PEMs) structure was observed and globular embryos could be differentiated from the C1-type calli, whereas in the other five types of calli (C2-C6), the disintegration of PEMs structure, a large number of irregularly shaped cells and degraded cells were observed, there were no differentiation of globular embryos from the these 5 types of calli. The results showed that the contents of soluble sugar and starch were the highest in the calli with high embryonic potential, high activity of POD appeared in highly embryonic potential calli and browned calli, the activity of superoxide SOD was lower in the calli with high embryonic potential and the calli with differentiated globular embryos, while the a

关 键 词：红松 长期继代 胚性愈伤组织 形态特征 生理指标 

分 类 号：S722[农业科学—林木遗传育种] Q813[农业科学—林学]