Zhao Xupeng, Luo Keming, Zhou Yue, Wu Xiuhua, Yang Li, Tang Shaohu
Sheng Wu Gong Cheng Xue Bao. 2013 Nov;29(11):1599-606.
A high efficient in vitro regeneration protocol was developed from leaf explants of the female 'Red Sun' kiwifruit (Actinidia chinensis) and the multiplication coefficient and rooting rate of adventitious buds were also optimized. This method does not require formation of callus tissues which leads to somaclonal variations. The results show that the adventitious buds developing directly from explants tissue were noticed after 30 d of culture. The maximum regeneration frequency of adventitious buds is 100% and 18.67 shoots was observed in each leaf explants when MS medium was supplemented with 3.0 mg/L BA+1.0 mg/L NAA. The optimal culture medium for bud multiplication is MS+2.0 mg/L BA+1.0 mg/L NAA+0.1 mg/L GA3 and the multiplication coefficient reached 8.63. On the rooting medium with 1/2 MS+0.8 mg/L IBA for 15 d, the adventitious plantlets were transferred into matrix perlite supplied with 1/2 MS liquid medium for 15 d and the rooting rate reached 100%. 95 out of 98 plantlets (96.94%) survived acclimatization, producing healthy plants in the greenhouse. Taken together, a highly efficient regeneration method via leaf explants of 'Red Sun' kiwifruit was successfully established. This protocol may be useful for micropropagation and genetic transformation studies of 'Red Sun' kiwifruit.
以雌性‘红阳’猕猴桃(中华猕猴桃)叶片为外植体,建立了高效的离体再生体系,并对不定芽的增殖系数和生根率进行了优化。该方法无需形成愈伤组织,从而避免了体细胞无性系变异。结果表明,培养30 d后可观察到外植体组织直接产生不定芽。当MS培养基添加3.0 mg/L BA + 1.0 mg/L NAA时,不定芽的最大再生频率为100%,每个叶片外植体可观察到18.67个芽。芽增殖的最佳培养基为MS + 2.0 mg/L BA + 1.0 mg/L NAA + 0.1 mg/L GA3,增殖系数达到8.63。在含有1/2 MS + 0.8 mg/L IBA的生根培养基上培养15 d后,将不定芽苗转移到添加1/2 MS液体培养基的珍珠岩基质中培养15 d,生根率达到100%。98株苗中有95株(96.94%)驯化成活,在温室中长成健康植株。综上所述,成功建立了一种通过‘红阳’猕猴桃叶片外植体的高效再生方法。该体系可能对‘红阳’猕猴桃的微繁殖和遗传转化研究具有重要意义。