氨基酸对荔枝愈伤组织增殖及体胚发生体系优化的研究  

Study on the optimization of amino acids on proliferation of calli and somatic embryogenesis in Litchi chinensis‘Feizixiao'

作　　者：王果[1] 刘耀婷 王家保[1] 李焕苓[1] WANG Guo;LIU Yaoting;WANG Jiabao;LI Huanling(Environments and Plant Protection Institute,Chinese Academy of Tropical Agriculture Sciences/Danzhou Scientific Observing and Ex-perimental Station of Agro-Environment,Ministry of Agriculture and Rural Affairs,P.R.China,Danzhou 571737,Hainan,China;Col-lege 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》2023年第11期2466-2476,共11页Journal of Fruit Science

基　　金：国家重点研发计划(2019YFD1000900);财政部和农业农村部:国家现代农业产业技术体系资助(CARS-32)。

摘　　要：【目的】探究氨基酸对荔枝愈伤组织增殖及体胚诱导的影响。【方法】采用L9(34)正交设计,分别在妃子笑荔枝愈伤组织继代及体胚诱导阶段添加不同氨基酸,比较其对愈伤组织增殖及体胚诱导的影响。【结果】在愈伤组织增殖阶段培养基中添加氨基酸均显著提高愈伤组织增殖率,最优处理为基础继代培养基添加0.1 g·L^(-1)γ-氨基丁酸+0.2 g·L^(-1)谷氨酰胺,愈伤组织增殖率为初始接种量的14.04倍。与对照相比,氨基酸处理组的胚性愈伤组织淡黄,颗粒较小,细胞呈椭圆形,细胞质浓厚,多为正在分裂状态;非胚性愈伤组织浅白,水渍严重,着生于顶部或者边缘,易于剔除。在愈伤组织增殖阶段培养基中添加γ-氨基丁酸、谷氨酰胺和丙氨酸对后续的体胚发生及萌发影响极显著(p<0.01),最优配方为基础继代培养基添加0.3 g·L^(-1)γ-氨基丁酸+0.4 g·L^(-1)谷氨酰胺+0.05 g·L^(-1)丙氨酸,每克愈伤组织可获得461个体胚,77株组培苗。在体胚发生阶段培养基中添加氨基酸,获得的体胚发生及萌发最优配方为基础诱导培养基添加0.3 g·L^(-1)γ-氨基丁酸+0.4 g·L^(-1)谷氨酰胺+0.05 g·L^(-1)丙氨酸,每克愈伤组织可获得270个体胚,72株组培苗。【结论】氨基酸可调节愈伤组织胚性,提高愈伤组织增殖率,促进体胚发生及萌发。【Objective】Clonal propagation is an essential tool in biotechnology and breeding programs.Among the available clonal propagation methods,somatic embryogenesis is the most attractive technique for the large-scale propagation of plants and the long-term conservation of different embryogenic cell lines without changing their initial characteristics.Additionally,somatic embryogenesis is an ideal system for cell biology and molecular studies,including the study of the cell cycle,cell division,cell differentiation,and for understanding different genetic and epigenetic mechanisms for gene expression and phenotypic variation in plants.However,for many species,in vitro clonal propagation is currently difficult or inefficient.The low efficiency of somatic embryo induction is among the major problems hampering efficient regeneration and limiting practical applications.Therefore,high-quality somatic embryo production is one of the most important factors for using somatic embryogenesis protocols in commercial propagation and breeding programs.Litchi(Litchi chinensis Sonn.)is one of the treasure fruits cultivated in the south of China.Breeding elite litchi varieties with resistance to diseases is very important to the litchi industry.Many studies on the somatic embryogenesis of litchi have been reported.However,there has been no detailed report on the the effect of amino acids on callus proliferation and somatic embryo induction in litchi.The study aimed to investigate the effects of amino acids on the efficiency of somatic embryogenesis in Feizixiao litchi and apply these findings to improve the protocol for the induction of somatic embryos.【Methods】The calli Feizixiao litchi were used as materials,the effects of different amino acids on callus proliferation,somatic embryogenesis and regeneration were studied using an L9(34)orthogonal experiment.【Results】The results showed that the effects of glutamine(Gln)andγ-aminobutyric acid(GABA)on the callus morphology and proliferation rate were significant(p0.01),while th

关 键 词：荔枝 愈伤组织 体胚发生 体胚萌发 氨基酸 

分 类 号：S667.1[农业科学—果树学]