刺梨茎尖的玻璃化超低温保存及植株再生Cryopreservation of in vitro Rosa roxburghii shoot-tips by vitrification and its regeneration
陈红,程秀枝,林田
摘要(Abstract):
【目的】建立刺梨茎尖超低温保存技术体系,为刺梨种质资源的长期稳定保存提供新途径。【方法】以刺梨为材料,研究了适合茎尖超低温保存的蔗糖预培养时间与浓度、预处理时间、玻璃化液处理时间等影响因素。【结果】适合超低温保存芽诱导的适宜培养基为MS+0.5 mg·L-16-BA+30 g·L-1蔗糖+7 g·L-1琼脂。刺梨茎尖玻璃化法超低温保存体系为:约0.5 cm长的腋芽茎尖在4℃条件下于培养基MS+0.5 mg·L-16-BA+0.05 mg·L-1IAA+0.3 mo1·L-1蔗糖+7 g·L-1琼脂上预培养3 d,切取约2 mm长的茎尖,室温下LS装载液处理40~50 min,然后用玻璃化液PVS2于0℃下处理40 min,换入新鲜的PVS2,迅速投入液氮中。保存24 h后,在40℃水浴中化冻,用MS+1.2 mol·L-1蔗糖+100 mg·L-1Vc+0.5 mg·L-1GA3培养基洗涤2次,接种到MS+0.3 mol·L-1蔗糖的培养基上暗培养1 d,再转接到MS+0.1mg·L-16-BA+0.05 mg·L-1IAA+30 g·L-1蔗糖+7.0 g·L-1琼脂上暗培养7 d后转入正常光下培养,成活率可达44.3%。【结论】建立了刺梨茎尖超低温保存技术体系。
关键词(KeyWords): 刺梨;茎尖;超低温保存;玻璃化法
基金项目(Foundation): 贵州省科技攻关项目(黔科合农G字[2009]4003)
作者(Author): 陈红,程秀枝,林田
DOI: 10.13925/j.cnki.gsxb.2012.06.033
参考文献(References):
- [1]FANG Xiu-gui,LI Si-biao.ZHENG Yi-qing.The nutritive value ofRosa roxburghii Tratt and itsdevelopment and utilization[J].Scienceand Technology of Food Industry,2004,25(1):137-138.方修贵,李嗣彪,郑益清.刺梨的营养价值及其开发利用[J].食品工业科技,2004,25(1):137-138.
- [2]NIINO T,TASHIRO K,SUZUKI M,OHUCHI S,MAGOSHI J,AKIHAMA T.Cryopreservation of in vitro grown shoot tips of cherryand sweet cherry by one-step vitrification[J].Scientia Horticulturae,1997,70:155-163.
- [3]WANG Zi-cheng,DENG Xiu-xin.Cyopreservation of citrus shoot-tips by vitrification and regeneration[J].Acta Horticulturae Sinica,2001,28(4):301-306.王子成,邓秀新.玻璃化法超低温保存柑桔茎尖及植株再生[J].园艺学报,2001,28(4):301-306.
- [4]MATSUMOTO T,MOCHIDA K,ITAMURA H,SAKAI A.Cryop-reservation of persimmon(Diospyros kaki Thunb.)by vitrification ofdormant shoot tips[J].Plant Cell Rep,2001,20(5):398-402.
- [5]ZHAO Yan-hua,WU Yong-jie.Study on cryopreservation of in vitroculturedcabemet france shoot tips[J].Acta Horticulturae Sinica,2001,28(1):62-64.赵艳华,吴永杰.‘品丽珠’葡萄离体茎尖超低温保存的研究[J].园艺学报,2001,28(1):62-64.
- [6]AI Peng-fei,LUO Zheng-rong.Cryopreservation of dormant shoot-tips of persimmon by vitrification and plant regeneration[J].ScientiaAgriculturae Sinica,2003,36(5):553-556.艾鹏飞,罗正荣.柿休眠芽茎尖玻璃化法超低温保存及植株再生[J].中国农业科学,2003,36(5):553-556.
- [7]MATSUMOTO T,SAKAI A.Cryopreservation of axillary shoot tips ofin vitro-grown grape(Vitis)by a two-step vitrification protocol[J].Euphytica,2003,131(3):299-304.
- [8]ZENG Ji-wu,YI Gan-jun,ZHANG Qiu-ming.Cryopreservation ofin vitro papaya shoot-tips by vitrification technique and its regener-ation[J].Acta Horticulturae Sinica,2004,31(1):29-33.曾继吾,易干军,张秋明.番木瓜茎尖的玻璃化法超低温保存及其植株再生[J].园艺学报,2004,31(1):29-33.
- [9]XU Xiao-biao,GU Qing-qing,CAI Zu-guo,DENG Xiao-mei,ZHANG Qiu-ming.Cryopreservation of in vitro cultured kiwifruitshoot-tips by vitrification and their regeneration[J].Scientia Agri-culturae Sinica,2006,33(4):842-844.徐小彪,辜青青,蔡祖国,邓小梅,张秋明.玻璃化法超低温保存猕猴桃离体茎尖及其植株再生[J].园艺学报,2006,33(4):842-844.
- [10]TSAI S F,YEH S D,CHAN C F,LIAW S I.High-efficiency vitrifi-cation protocols for cryopreservation of in vitro grown shoot tips oftransgenic papaya lines[J].Plant Cell Tiss Organ Cult,2009,98(2):157-164.
- [11]ZHAO Yan-hua,WU Ya-qin,CHENG He-he,LI Chun-min,CHENXiao-ling,LU Xin-xiong.Cryopreservation of shoot tips from Prunus[J].Scientia Agriculturae Sinica,2008,35(3):423-426.赵艳华,吴雅琴,程和禾,李春敏,陈晓玲,卢新雄.李离体茎尖的超低温保存[J].园艺学报,2008,35(3):423-426.
- [12]AI Peng-fei,LUO Zheng-rong.Cryopreservation of in vitro shoot-tips of persimmon and date plum by vitrification and genetic stabilityof regenerated plantlets[J].Scientia Agricultura Sinica,2004,37(12):2023-2027.艾鹏飞,罗正荣.柿和君迁子试管苗茎尖玻璃化法超低温保存及再生植株遗传稳定性研究[J].中国农业科学,2004,37(12):2023-2027.
- [13]ZHAO Yan-hua,WU Ya-qin.Cryopreservation of shoot tips frompeach and its regeneration[J].Scientia Agriculturae Sinica,2006,33(5):1042-1044.赵艳华,吴雅琴.桃离体茎尖的超低温保存及植株再生[J].园艺学报,2006,33(5):1042-1044.