‘红阳’猕猴桃叶盘高频直接再生体系的建立

Establishment of high frequency regeneration via leaf explants of ‘Red Sun’ kiwifruit (Actinidia chinensis)

摘要:

以‘红阳’猕猴桃雌株幼叶为外植体，直接诱导产生不定芽，并对不定芽增殖以及生根体系进行优化，建立了高频再生体系。结果表明，在MS+3.0 mg/L BA+1.0 mg/L NAA培养基中，不定芽诱导率达100％，平均出芽数达18.67个/叶盘；在MS+2.0 mg/L BA+1.0 mg/L NAA+0.1 mg/L GA3培养基中，增殖率达100％，1~6代不定芽平均繁殖系数达8.63；不定芽在1/2 MS+0.8 mg/L IBA培养基中培养15 d，然后继代至含1/2 MS液体培养基的珍珠岩中培养15 d，生根率达100％，且其根系发育良好；98株试管苗移栽到土壤盆钵中，成活95株，成活率达96.94％。本试验成功建立了红阳猕猴桃叶片高频直接再生体系，该体系诱导产生不定芽周期短，出芽率高，不定芽增殖系数大，生根率高且根系发达，为红阳猕猴桃试管苗的工厂化生产和遗传转化奠定了基础。

Abstract:

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.