成年态柑橘枝条外植体无菌化培养研究  

Study on the aseptic culture of mature stem explants of citrus

作　　者：张平[1] 刘娟[1] 胡其军 张玉婷 孔佑涵[1] 吴娟娟[1] ZHANG Ping;LIU Juan;HU Qijun;ZHANG Yuting;KONG Youhan;WU Juanjuan(Hunan Horticulture Research Institute/Huazhong Experimental Station of Scientific Observation of Fruit/Hunan Regional Characteristic Germplasm Resource Repository for Fruit and Flower,Changsha,Hunan,410125,China)

机构地区：[1]湖南省园艺研究所/国家农业部华中地区果树科学观测实验站/湖南省地方特色果树花卉资源圃,长沙410125

出　　处：《中国南方果树》2025年第2期52-55,59,共5页South China Fruits

基　　金：湖南省农业科学院科技创新项目(2022CX46);湖南省农业资金项目(湘财农指[2022]23号,湘财农指[2023]43号)资助。

摘　　要：为了筛选成年态(结果树)柑橘枝条外植体无菌化处理方案,进而提高柑橘离体组织培养效率,在冰糖橙、卡拉卡拉脐橙、塔罗科血橙、大分4号温州蜜柑、太田椪柑和菊花芯柚结果树上取当年生半木质化枝条,剪截枝段为外植体,在75%乙醇浸泡30 s基础上,进行次氯酸钠(NaClO)和特美汀(Timentin)单独或组合处理,比较不同无菌化处理条件下培养效果,并取无菌化培养萌发腋芽进行茎尖嫁接试验。结果表明,Timentin处理浓度≤500 mg/L时,对成年态柑橘枝条外植体的腋芽生长影响不大;浓度2%NaClO处理后,外植体在培养过程中污染率变化不大;75%乙醇浸泡30 s+2%NaClO浸泡20 min+300 mg/L Timentin的处理条件下,冰糖橙外植体培养14 d时的污染率为14.4%,无菌化培养出的腋芽进行茎尖嫁接的成活率能达到35.6%以上。75%乙醇浸泡30 s+2%NaClO浸泡20 min+300 mg/L Timentin用于成年态柑橘枝条外植体无菌化培养,污染率低,腋芽活力高,能满足柑橘良种脱毒等工程的需要。In order to screen the sterilization protocols for mature(fruit-bearing)stem explants of citrus and improve the efficiency of in vitro tissue culture of citrus,current-year semi-lignified branches were collected from fruit-bearing trees of Bingtang orange,Cara Cara navel orange,Tarocco blood orange,Oitakaken 4GO Satsuma mandarin,Ota Ponkan,and Juhuaxin pomelo and cut into segment as explant.After immersed in 75%ethanol for 30 seconds,the explants were treated separately or in combination with sodium hypochlorite(NaClO)and Timentin to compare the effects of different sterilization treatments on culture.Axillary buds generated from sterile cultures were used to conduct shoot-tip grafting experiments.The results showed that Timentin treatment at concentration≤500 mg/L had little negative impact on the axillary bud growth of the explants.After 2%NaClO treatment,the contamination rate of the explants showed no significant change during the culture.Under the conditions of immersion for 30 s in 75%ethanol,then immersion for 20 min in 2%NaClO+300 mg/L Timentin,the bacterial contamination rate of the Bingtang orange explants cultured for 14 days was only 14.4%.The survival rate of the shoot-tip grafted axillary buds cultured from explants was more than 35.6%.The combined treatment of 75%ethanol immersion for 30 s+immersion for 20 min with 2%NaClO+300 mg/L Timentin had low contamination rates and high axillary bud vigor for sterile culture of mature stem explants of citrus,which can meet the requirements for virus-free citrus breeding programs.

关 键 词：柑橘 成年态外植体 污染率 茎尖嫁接 

分 类 号：S666[农业科学—果树学]