激素调控对椰枣体胚发生的生理响应及体系优化  

Physiological Response and System Optimization of Hormone Regula-tion to Somatic Embryogenesis of Date Palm

作　　者：张宁 张婷芊 韩宇晶 张馨月 杨力颖 符海泉[1] 胡伟 ZHANG Ning;ZHANG Tingqian;HAN Yujing;ZHANG Xinyue;YANG Liying;FU Haiquan;HU Wei(Coconut Research Institute,Chinese Academy of Tropical Agricultural Sciences/National Tropical Palm Germplasm Reserve,Wenchang,Hainan 571300,China;College of Horticulture,Nanjing Agricultural University,Nanjing,Jiangsu 210095,China;College of Tropical Crops,Yunnan Agricultural University,Pu’er,Yunnan 665000,China;Yunnan Xinxing Greening Engineering Co.,Ltd.,Kunming,Yunnan 650031,China)

机构地区：[1]中国热带农业科学院椰子研究所/国家热带棕榈种质资源圃,海南文昌571300 [2]南京农业大学园艺学院,江苏南京210095 [3]云南农业大学热带作物学院,云南普洱665000 [4]云南新兴绿化工程有限公司,云南昆明650031

出　　处：《热带作物学报》2025年第2期333-342,共10页Chinese Journal of Tropical Crops

基　　金：海南省自然科学基金项目(No.323QN271);云南省重大科技专项计划项目(No.202202AE090104);中央级公益性科研院所基本科研业务费专项(No.1630152024003)。

摘　　要：椰枣为棕榈科作物,雌雄异株,基因高度杂合,性状分离严重。种苗繁育依赖于无性繁殖,组织培养技术是目前实现椰枣种苗商业化生产的主要手段。但是,体胚发生困难、诱导周期长导致体系构建困难、组培成本高,研究激素调控对椰枣体胚内源代谢物质的影响,优化体胚发生体系成为当务之急。本研究基于前期对椰枣外植体种类的筛选、愈伤组织及胚性愈伤组织诱导的基础上,利用正交试验设计,通过对MS培养基中植物生长调节剂种类和浓度进行筛选配比,从愈伤组织中诱导出体胚,计算体胚发生率、诱导周期及成活率,对不同激素水平下获得的椰枣体胚进行取样,观察体胚的形态变化规律;测定样品中总蛋白(Cpr)、可溶性糖(WSS)、过氧化氢还原酶(CAT)、谷胱甘肽还原酶(GR)、氨基酸(AA)、生长素(auxin)、细胞分裂素(CTK)、脱落酸(ABA)、赤霉素(GA)等指标的含量变化规律。结果表明:C_(5)(MS+30 g/L蔗糖+2 mg/L NAA+2 mg/L 6-BA+3 mg/L 2,4-D+3 mg/L 2-ip)处理的体胚发生时间最短,在C_(9)(MS+30 g/L蔗糖+3 mg/L NAA+3 mg/L 6-BA+2 mg/L 2,4-D+3 mg/L 2-ip)处理下,体胚中过氧化氢酶含量和氨基酸含量最高,在C5处理下,蛋白质含量最高;变异系数最大的为谷胱甘肽还原酶含量,极差最大的是氨基酸含量;相关性分析表明,细胞分裂素和生长素均与谷胱甘肽还原酶相关性最强,可溶性糖与蛋白质和氨基酸的相关性最强;主成分分析显示,前4个主成分累计贡献率达到88.714%,第1个主成分的特征值为3.877,贡献率为43.076%,在第1个主成分中,主要由生长素含量、可溶性糖含量、蛋白质含量及赤霉素含量这4个指标共同影响,其中生长素含量的特征值最大为0.921,主要反映生长素对体胚发生的影响。本研究结果为优化椰枣体胚发生体系提供理论依据,同时为实现椰枣高效快繁奠定技术基础。Date palm is hermaphroditic and highly heterogeneous in genetics,resulting in severe phenotypic variation.Propagation of seedlings depends on asexual reproduction,and tissue culture technology is the main means of commer-cializing date seedling production at present.However,the difficulty in inducing embryogenesis and the long induction cycle lead to difficulties in system construction and high tissue culture costs.Therefore,it is urgent to study the effects of hormone regulation on the endogenous metabolic substances of date embryos and optimize the embryogenesis system.This study is based on the selection of date explants,induction of callus and embryogenic callus in the previous studies,and uses orthogonal experimental design to screen and match the plant growth regulators and concentrations in the MS culture medium to induce embryogenesis from callus.The study statistically recorded the embryogenesis rate,induction cycle,and survival rate;date embryos with different levels of hormone were sampled;the morphological changes of date embryos in a regular manner were observed;and the changes in the content of total protein(Cpr),soluble sugar(WSS),catalase(CAT),glutathione reductase(GR),amino acids(AA),auxin,cytokinin(CTK),abscisic acid(ABA),and gibberellin(GA)were measured.The results showed that the embryogenic rate of C_(9) was higher than that of C_(5),C_(4),C_(6),and the time for embryogenesis was the shortest in the C_(5) treatment.In the C_(9) treatment,the highest content of cata-lase and amino acids was found in the embryos,and the highest protein content was found in the C_(5) treatment.The co-efficient of variation was the largest for glutathione reductase content,and the maximum difference was found in amino acid content.The correlation analysis showed that thiocyanate and gibberellin had the strongest correlation with glu-tathione reductase,and soluble sugar had the strongest correlation with protein and amino acid.Principal component analysis showed that the first four principal components accounted f

关 键 词：椰枣 体胚 激素 愈伤组织 

分 类 号：S667.9[农业科学—果树学]