摘要
【目的】探明百蕊草体细胞胚发生过程中的组织细胞特征及内源生长调节物质的变化规律,为建立百蕊草体细胞胚组培苗快繁技术体系提供理论参考,并为丰富百蕊草细胞分化和器官形态建成等基础理论提供基础信息。【方法】采用显微观察方法对叶片外植体诱导的百蕊草愈伤组织及不同分化培养时期的体细胞胚进行组织细胞学观察,并检测不同分化时期的体细胞胚内源激素和类似物含量、抗氧化酶活性及可溶性蛋白质含量。【结果】胚性和非胚性愈伤组织的形态和细胞结构差异明显。体细胞胚分化经历的时期有多细胞原胚(PM)、球形胚(GE)、心形胚(HE)、鱼雷形胚(TE)和子叶胚(CE)等。内源激素及类似物中反式玉米素(tZT)含量与玉米素核苷(ZR)和吲哚-3-乙酸(IAA)含量呈显著正相关(r=0.421^(*),r=0.536^(*)),与脱落酸(ABA)含量呈极显著负相关(r=-0.688^(**)),tZT与IAA、ZR对百蕊草体细胞胚发生的作用相同。ABA含量与IAA和ZR含量呈显著负相关(r=-0.401^(*),r=-0.430^(*))。赤霉素3(GA_(3))含量与IAA含量呈弱正相关性(r=0.313),作用类似,均可促进细胞和器官的伸长。水杨酸(SA)含量变化呈波浪形变化,且最高值4.56 ng/g和最低值3.29 ng/g分别出现在PM和GE时期,说明与体细胞胚发生有关。抗氧化酶中过氧化物酶(POD)和过氧化氢酶(CAT)活性持续升高,并且保持在较高水平,尤其是在胚分化后期。超氧化物歧化酶(SOD)活性从PM至GE时期迅速上升并于GE出现峰值29.19 U/(g·min)之后下降,但其活性较POD和CAT低。可溶性蛋白(SP)含量由PM至GE时期稍有下降,于HE时期升高达最大值2.24 ng/g,之后缓慢降低。SP含量呈波动性变化。【结论】百蕊草体细胞胚的发育过程与合子胚的发育过程相似,并且内源激素及类似物异戊烯腺嘌呤(IP)、tZT、ZR、IAA、GA 3、ABA、SA及抗氧化酶(POD、CAT、SOD)和SP具有不同程度的影响和调控作用�
【Objective】The characteristics of tissue cell and the changes in endogenous growth regulators during the process of somatic embryogenesis in Thesium chinense were explored,which provided the theoretical reference for the establishment of a rapid propagation technology system for tissue culture seedlings of T.chinense somatic embryos,as well as provided the basic information for enriching the basic theoretical of cell differentiation and organ morphogenesis in T.chinense.【Method】The microscopic observation method was used to observe the tissue and cytology of callus induced by leaf explants and somatic embryos at different differentiation and culture stages.The content of endogenous hormones and analogues,antioxidant enzyme activity and soluble protein content of somatic embryos at different differentiation stages were detected.【Result】There were significant differences in the morphology and cellular structure between embryonic and non embryonic callus.The differentiation of somatic embryos underwent stages such as multicellular proembryo(PM),globular embryo(GE),heart-shaped embryo(HE),torpedo-shaped embryo(TE),and cotyledon embryo(CE).The content of transzeatin(tZT)in endogenous hormones and analogues was significantly and positively correlated with the content of zeatin nucleoside(ZR)and indole-3-acetic acid(IAA)(r=0.421^(*),r=0.536^(*)),and extremely significantly and negatively correlated with the content of abscisic acid(ABA)(r=-0.688^(**)).tZT,IAA and ZR had the same effect on the somatic embryogenesis of T.chinense.There was a significant and negative correlation between ABA content and IAA and ZR content(r=-0.401^(*),r=-0.430^(*)).The content of gibberellin 3(GA_(3))and IAA showed a weak positive correlation(r=0.313),and their effects were similar,both could promote cell and organ elongation.The changes in salicylic acid(SA)content showed a wavy pattern,with the highest value of 4.56 ng/g and the lowest value of 3.29 ng/g occurring during PM and GE periods,respectively,indicating a correlation
作者
黄燕芬
吴坤
张文香
郁琴红
田海浪
HUANG Yanfen;WU Kun;ZHANG Wenxiang;YU Qinhong;TIAN Hailang(School of Biological Sciences,Guizhou Education University,Guiyang,Guizhou 550018;Key Laboratory of Guizhou Bio-resource Development and Utilization,Guiyang,Guizhou 550018;School of Chemistry and Material Sciences,Guizhou Education University,Guiyang,Guizhou 550018;College of Life Sciences,Hengshui University,Hengshui,Hebei 053000,China)
出处
《贵州农业科学》
CAS
2023年第11期98-108,共11页
Guizhou Agricultural Sciences
基金
贵州省科技厅自然科学基金项目“百蕊草体细胞胚发生系统生理生化机理及其在培育再生体系上应用的研究”[黔科合基(2018)1117]。
关键词
百蕊草
体细胞胚
组织细胞学
内源激素
抗氧化酶
可溶性蛋白
Thesium chinense
somatic embryo
histocytology
endogenous hormones
antioxidant enzymes
soluble protein
