‘阳光玫瑰’葡萄组培脱毒快繁技术研究  

Study on rapid propagation technology of virus-free seedlings by tissue culture in‘Shine Muscat’grape

作　　者：林茜[1,2] 高营营 覃换玲 黄天琨 赵宇 王钟霞 陈淑媛 LIN Qian;GAO Yingying;QIN Huanling;HUANG Tiankun;ZHAO Yu;WANG Zhongxia;CHEN Shuyuan(Biotechnology Reserach Institute,Guangxi Academy of Agricultural Sciences,Nanning 530007,Guangxi,China;Guangxi Plant Tissue Culture Seedling Co.LTD,Nanning 530007,Guangxi,China)

机构地区：[1]广西农业科学院生物技术研究所,南宁530007 [2]广西植物组培苗有限公司,南宁530007

出　　处：《果树学报》2021年第3期435-443,共9页Journal of Fruit Science

摘　　要：【目的】培育‘阳光玫瑰’葡萄组培脱毒苗木,初步建立‘阳光玫瑰’葡萄组培脱毒快繁技术体系。【方法】采用MS为基本培养基,以植物生长调节剂6-BA、NAA和IBA为变量,接种后‘阳光玫瑰’葡萄组培苗的生长状况为因变量;通过热处理结合茎尖培养技术,在32℃预热处理7 d,再逐渐升温至37℃热处理30 d后,剥取茎尖进行培养,待获得完整植株时,利用RT-PCR检测方法对‘阳光玫瑰’葡萄组培苗进行病毒检测。【结果】‘阳光玫瑰’葡萄嫩茎段外植体经75%乙醇30 s+0.1%氯化汞8 min处理,外植体的污染率和褐化率最低;经消毒灭菌的外植体接种到添加含有6-BA和NAA的MS培养基上诱导萌发,在1.5 mg·L^(-1)6-BA和0.2 mg·L^(-1)NAA的培养基上萌芽率最高;将启动培养获得的无菌新芽,接种到含有6-BA和NAA的MS培养基中进行继代培养,在1.0 mg·L^(-1)6-BA+0.1 mg·L^(-1)NAA的培养基中单芽增殖效果最明显;把继代培养中生长健壮的单芽切下,转入添加IBA和NAA的1/2 MS培养基中进行生根培养,在添加0.4 mg·L^(-1)IBA和0.2 mg·L^(-1)NAA的1/2 MS培养基上生根效果最佳;采用热处理结合茎尖培养进行‘阳光玫瑰’葡萄组培苗的脱毒处理,热处理植株的成活率为78%,茎尖成活率为60%,经检测,再生植株不带葡萄卷叶病毒1(GLRaV-1)、葡萄卷叶病毒3(GLRaV-3)、葡萄病毒A(GVA)、葡萄斑点病毒(GFkV)、葡萄扇叶病毒(GFLV)。【结论】初步建立了‘阳光玫瑰’葡萄组培脱毒快繁技术体系,为‘阳光玫瑰’葡萄组培脱毒苗的工厂化生产提供了技术支撑。【Objective】‘Shine Muscat’grape,with its unique advantages of seedless,high sugar content,rich rose fragrance and good flavor,surpasses the quality of other grape varieties and is welcomed by consumers.In recent years,‘Shine Muscat’grape has been widely introduced and planted in Zhejiang,Jiangsu,Anhui,Yunnan,Guangxi and other places,showing a broad market prospect.With the development of grape industry,the demand for‘Shine Muscat’grape seedlings is growing rapidly.However,in the cultivation of this variety in China,the plants generally show the symptoms of viral diseases,so that the yield and berry quality were seriously affected.The traditional way of grape seedling production to remove the virus is inefficient.Therefore,it is necessary to establish a rapid propagation technology system for virus-free seedlings of‘Shine Muscat’grape by combining tissue culture with heat treatment method.【Methods】Based on the heat treatment method,the virus-free zone of the stem tip can expand by inactivating the virus,and the stem tip has the advantage of taking small apical meristems to grow a whole plant.In this paper,the tissue culture seedlings of‘Shine Muscat’grape were treated by combining heat treatment with stem tip culture.Firstly,the stem segments with axillary bud area of new canes were used as explants,and they were disinfected by different disinfection methods.Then they were inoculated on MS medium containing different concentrations of plant growth regulators.After a few days,the pollution rate,survival rate,germination rate,subculture multiplication coefficient,rooting rate,root length and root number of explants were calculated.The best method of disinfection,disinfection time,start medium,subculture medium,rooting medium for explants was determined.Secondly,when robust tissue culture seedlings were obtained,they were heat-treated at different culture temperatures and durations.At the end of treatment,the stem tips were selected and cultured to grow into intact plants.Finally,the tube s

关 键 词：‘阳光玫瑰’葡萄 组织培养 快繁 脱毒 

分 类 号：S663.1[农业科学—果树学]