影响枣果肉离体愈伤组织cAMP含量的因素  

Factors affecting cAMP content in jujube pulp callus

作　　者：郭晓雪 赵若雨 李瑞梅 朱文慧 杜丹 赵佳雪 代丽[1] 刘志国 刘孟军[1] 王立新 GUO Xiaoxue;ZHAO Ruoyu;LI Ruimei;ZHU Wenhui;DU Dan;ZHAO Jiaxue;DAI Li;LIU Zhiguo;LIU Mengjun;WANG Lixin(College of Horticulture,Hebei Agricultural University,Baoding 071001)

机构地区：[1]河北农业大学园艺学院,保定071001

出　　处：《中国果树》2024年第1期70-77,共8页China Fruits

基　　金：河北省引进留学人员资助项目(C20210114);河北省教育厅科学技术研究项目资助(QN2022017);河北省省属高校基本科研业务费研究项目(KY2021059);国家自然科学基金青年科学基金项目(32201594);财政部和农业农村部:国家现代农业(桃)产业技术体系(CARS-30-2-07)。

摘　　要：环磷酸腺苷(cAMP)具有第二信使功能及重要的药用价值,但含量通常极低,通过组织培养快速繁殖富含cAMP的植物材料用于提取cAMP前景广阔。枣树原产于我国,其成熟果肉中富含cAMP。为探明利用枣果肉愈伤组织生产cAMP的可行性,研究了基因型和果实成熟度对果肉愈伤组织cAMP含量的影响以及培养基类型、继代次数和cAMP合成关键酶腺苷酸环化酶(AC)的辅离子等对果肉愈伤组织cAMP含量的调控作用。选取不同枣果实以及相对应的不同基因型的枣果肉愈伤组织、不同培养基上的果肉愈伤组织、不同发育阶段及不同继代次数的果肉愈伤组织、经AC辅离子处理过的果肉愈伤组织,利用ELISA试剂盒测定其cAMP含量。结果表明,幼果期果肉愈伤组织中cAMP含量高于其幼果期果实中的cAMP含量,不同基因型果肉愈伤组织中cAMP含量存在差异,但远小于不同基因型果肉中cAMP含量的差异。幼果期的果肉最容易形成愈伤组织,随着果实成熟度的增加越来越难以诱导出愈伤组织,而不同发育阶段果实诱导产生的愈伤组织中cAMP含量的差异不大,远小于不同发育阶段果肉中cAMP含量的差异。培养基类型不仅影响愈伤组织的形成,还显著影响果肉愈伤组织中cAMP的含量;继代10次的果肉愈伤组织cAMP含量低于继代5次的。培养基中添加不同浓度及不同处理时间段的AC辅离子(MgCl_(2)、Mn Cl_(2)、CaCl_(2))对果肉愈伤组织中cAMP含量有一定的影响,100μmol/L MnCl_(2)处理48 h可显著提高幼果期果肉愈伤组织的cAMP含量,100μmol/L CaCl_(2)处理48 h可显著提高小紫枣和酸枣2X半红期果肉愈伤组织的cAMP含量,100μmol/L MgCl_(2)处理48 h可显著提高稷山板枣半红期果肉愈伤组织的cAMP含量。枣果肉愈伤组织基因型、果实成熟度、培养基类型、继代次数、AC辅离子对枣果肉愈伤组织中cAMP含量有着一定的影响。因此,利用枣果肉愈伤组织培�Cyclic adenosine monophosphate(cAMP),low content in organisms,functions a second messenger which involves in physiological and biochemical regulation in plants.In addition,cAMP plays an important role in plant growth and development,environmental adaptation,and has certain medicinal values and other practical functions.However rapid propagation of cAMP-rich plant materials through tissue culture method can be used to extract cAMP.Chinese jujube originated from China and its mature fruits contains high level of cAMP compare to other plant.Thus,in order to explore whether it is possible to use the jujube fruit callus to extract cAMP instead of jujube mature fruits,the effects of different jujube genotypes and fruit maturity on the cAMP content in jujube pulp callus,and the regulating effects of culture medium type,the number of sub-cultivation and the actions of the key enzyme adenylate cyclase(AC)on the cAMP content in jujube pulp callus were investigated.A variety of different jujube genotypes and different developmental stages of jujube fruits were selected for callus induction and cultivation after screening the most suitable cultivating conditions.After the callus of different genotypes and different developmental stages were induced,subcultivation and proliferation were carried out.When the callus grew to a well status,the samples were collected to do the cAMP content test.Briefly,the samples were evenly ground with a frozen grinder,and dissolved with pure water.The ratio of material to liquid was 1∶5.Then,it was bathed in water bath at 60℃for 3 h,and centrifuged at the speed of 5000 rpm for 20 min,and the supernatant was filtrated by proper membrane for the determination of cAMP.In addition,for the ions of the key enzyme AC treatments,three kinds of callus materials with high,medium and low content of cAMP were selected to treat with different concentrations of MgCl_(2),MnCl_(2) and CaCl_(2)(20μmol/L,50μmol/L,100μmol/L),respectively.Samples were selected after treatment for 0 h,24 h,48 h and 72 h.The

关 键 词：环磷酸腺苷 枣 果肉愈伤组织 培养基 AC辅离子 

分 类 号：S665.1[农业科学—果树学]