柿和君迁子试管苗缓慢生长法保存及其遗传稳定性研究

Conservation of in Vitro Shoots of Persimmon and Date Plum by Slow Growth and Genetic Stability of Recovered Plantlets

对柿(Diospyros kaki Thunb.)和君迁子(D.lotus L.)试管苗缓慢生长法保存及其恢复生长植株的遗传稳定性进行了研究,结果表明:低温(6±1)℃和光照800 lx(12 h/d)的条件下,试管苗在添加有甘露醇20g/L或PP333 1.0 mg/L的MS(1/2N)+蔗糖20g/L+琼脂7 g/L+PVP 500 mg/L培养基上或在含有CPPU 0.2 mg,/L的1/2 MS(1/2N)+蔗糖15 g/L+琼脂g/L+PVP 500 mg/L培养基上保存18个月,平均存活率在90％以上;上述3种方法保存后恢复生长的植株,在核DNA含量和染色体数目上没有发生改变;RAPD分析时表明,只有在添加PP333的保存中,次郎3个单芽姊妹系扩增出的1827条谱带中,增加了3条新带,变异率为0.16％,君迁子3个单芽姊妹系扩增出的1736条谱带中,增加了1条新带,缺失了14条带,变异率为0.86％。该结果为柿属植物种质资源缓慢生长法保存的有效性和可行性提供了理论依据。

Studies on conservation of in vitro shoots of persimmon ( Diospyros kaki Thunb. ) and date plum ( D. lotus L. ) by slow growth and genetic stability of recovered plantlets were carried out. Results showed that 90% of in vitro shoots conserved on MS (1/2N) supplemented with 20 g/L sucrose, 7 g/L agar and 500 mg/L soluble PVP added with 20 g/L mannitol or 1. 0 mg/L PP333 respectively or on 1/2MS (1/2N) supplemented with 15 g/L sucrose, 7 g/L agar and 500 mg/L soluble PVP added with 0. 2 mg/L CPPU for 18 months at (6±1)℃ and 800 lx (12 h photoperiod) maintained viability. Those recovered plantlets conserved with 3 methods above were analyzed on cytological and molecular levels respectively. Result showed both nuclear DNA content and chromosome numbers maintained as normal. And only few shoots cultivated with PP333, in which existed 0. 16% variation in Jirou persimmon owing to 3 distinct new bands appearing in 1827 amplified bands and 0. 86% variation in Date plum for 1 distinct new band and absence of 14 bands existing in 1736 amplified bands when 3 single-bud sibling lines were analyzed by RAPD markers respectively. Thus, it suggested these procedures would be effective and viable to conserve Diospyros germplasm by slow growth as above.