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园艺学报 ›› 2004, Vol. 31 ›› Issue (4): 441-450.

• 研究论文 • 上一篇    下一篇

柿和君迁子试管苗缓慢生长法保存及其遗传稳定性研究

艾鹏飞;罗正荣   

  1. (华中农业大学园艺植物生物学教育部重点实验室,武汉430070)
  • 收稿日期:2003-10-14 修回日期:2003-12-29 出版日期:2004-08-25 发布日期:2004-08-25

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

Ai Pengfei and Luo Zhengrong   

  1. (Key Laboratory of Honicuhural Plant Biology,Ministry of Education,PRC,Huazhong Agricultural University, Wuhan 430070,China)
  • Received:2003-10-14 Revised:2003-12-29 Online:2004-08-25 Published:2004-08-25

摘要: 对柿(Diospyros kaki Thunb.)和君迁子(D.1otus 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20 g/L或PPⅢ 1.0 mg/L的MS(1/2N)+蔗糖20 g/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7 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PP 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l4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。

关键词: 柿, 君迁子, 试管苗, 缓慢生长法, 遗传稳定性, 种质保存

Abstract: Studies on conservation of in vitro shoots of persimmon (Diospyros kaki Thunb.)and date plum (D.1otus 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 L sucrose,7 g/L agar
and 500 mg/L soluble PVP added with 20 L mannitol or 1.0 nqIg/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 f0r l8 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.Resuh showed both Ha—clear DNA content and chromosome numbers maintained as norma1.And only few shoots cultivated with PP11,in which existed 0.1 6% variation in Jirou persimmon owing to 3 distinct new bands appearing in l 827 ampli—fled bands and 0.86% variation in Date plum for l distinct new band and absence of 14 bands existing in 1 736
amplified bands when 3 single—bud sibling lines were analyzed by RAPD markers respectively.Thus.it sugges—ted these procedures would be effective and viable to conserve Diospyros germ plasm by slow growth as above.

Key words: Persimmon, Date plum, In vitro shoot, Slow gro~_h, Genetic stability, Conservation of gennplasm

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