南高丛蓝莓‘奥尼尔’工厂化组培快繁技术研究

Tissue culture and rapid propagation in factory of southern highbush blueberry‘O’Neal’

为了建立南高丛蓝莓‘奥尼尔’工厂化组培快繁技术体系,该文以带腋芽的茎段为外植体,研究外植体灭菌、丛生芽诱导、丛生芽增殖培养、瓶内生根以及移栽驯化。结果表明:(1)外植体最佳灭菌方法为10%NaClO处理15 min,污染率降低至14.44%,诱导率达49.38%;同样添加1.5 mg·L^(-1)ZT,以WPM为基础培养基诱导出的丛生芽明显多于MS。(2)丛生芽增殖的最佳培养基为WPM+NAA 0.1 mg·L^(-1)+ZT 1.5 mg·L^(-1),接种60 d后增殖系数达8.6,长势良好;瓶内生根的最佳处理方式为先用500 mg·L^(-1)IBA浸蘸20 s,然后接种于WPM+IBA 0.2 mg·L^(-1)的生根培养基中,培养90 d后,生根率、有效生根率、生根数量和根长分别达到96.3%、96.3%、12.7条和43.3 mm,且根系较发达、粗壮。(3)生根苗在移栽基质Ⅰ(河沙∶蛭石∶珍珠岩=1∶1∶1)的移栽成活率最高,达92.22%。以上说明,外植体经10%NaClO处理15 min,利用培养基WPM+ZT 1.5 mg·L^(-1)和WPM+NAA 0.1 mg·L^(-1)+ZT 1.5 mg·L^(-1)分别进行丛生芽诱导及增殖;丛生芽经500 mg·L^(-1)IBA浸蘸20 s,再接种于WPM+IBA 0.2 mg·L^(-1)进行生根,生根苗在河沙∶蛭石∶珍珠岩=1∶1∶1的基质移栽驯化,是南高丛蓝莓‘奥尼尔’最适工厂化组培快繁技术体系。该研究结果为今后‘奥尼尔’工厂化种苗快繁奠定了基础。

In order to establish a rapid propagation system of southern highbush blueberry‘O’Neal’,the stem section with axillary bud were sterilized as explants,induction of shoot cluster,and proliferation of shoot cluster,test-tube rooting,transplanting and domestication were also studied.The results were as follows:(1)The best sterilization treatment was that the explant was treated by 10%NaClO with 15 min,and the contamination rate decreased to 14.44%and the induction rate reached 49.38%;The best medium for induction of shoot cluster was WPM+ZT 1.5 mg·L^(-1).(2)The optimum medium for proliferation of shoot cluster was WPM+NAA 0.1 mg·L^(-1)+ZT 1.5 mg·L^(-1),and the coefficient of propagation reached 8.6 after subcultured for 60 d;The best treatment for test-tube rooting was that the shoot cluster dipped in 500 mg·L^(-1)IBA for 20 s firstly,then cultivated them with the medium WPM+IBA 0.2 mg·L^(-1),the rooting rate and effective rooting rate all reached 96.3%,and the rooting number and length were 12.7 and 43.3 mm after cultured for 90 d,at the same time,and the root developed well.(3)The survival rate of rooted seedlings reached 92.22%after transplanted in MediaⅠ(river sand∶vermiculite∶perlite=1∶1∶1).Thus,the best rapid propagation system of southern highbush blueberry‘O’Neal’was that:the explant treated by 10%NaClO with 15 min,the shoot cluster was inducted and proliferated with WPM+ZT 1.5 mg·L^(-1)and WPM+NAA 0.1 mg·L^(-1)+ZT 1.5 mg·L^(-1)respectively;The shoot cluster dipped in 500 mg·L^(-1)IBA for 20 s and cultivated with WPM+IBA 0.2 mg·L^(-1),and rooted seedlings were transplanted in media(river sand∶vermiculite∶perlite=1∶1∶1).The results play a foundation for providing rapid propagation seedlings and factory production of‘O’Neal’in the future.