潘学军;李霞;张文娥;

• 1:贵州省果树工程技术研究中心
• 2:贵州大学农学院

摘要(Abstract)：

【目的】为阐明缓慢生长法离体保存野生毛葡萄试管苗的机制,【方法】以野生毛葡萄‘花溪-9’为试材,利用缓慢生长法成功离体保存了供试材料试管苗,并从叶片解剖结构和生理生化变化2个方面阐述了毛葡萄试管苗离体保存存活率与叶片结构和抗氧化酶活性间的关系。【结果】温度、光照和烯效唑(S3307)对野生毛葡萄离体保存效果均有影响,其影响顺序为S3307>温度>光照,且3因素两两之间互作效应明显。野生毛葡萄在低温(10±2)℃、光照10μmol.m-2.s-1(12 h.d-1)和添加0.1 mg.L-1S3307的1/2MS+0.05 mg.L-1IBA+0.1 mg.L-1IAA+3%蔗糖+0.5%琼脂粉的培养基上保存效果良好,存活率可达86.33%。离体保存存活率较高的试管苗叶绿素含量和膜质过氧化产物MDA含量较低,保护酶(SOD、CAT、POD)活性强,渗透调节物质可溶性蛋白含量高,叶肉细胞的紧密度提高,单位面积内细胞数量增多。【结论】适宜的离体保存方法增加了毛葡萄叶肉细胞的紧密度,提高了试管苗的抗氧化能力,这可能是缓慢生长法离体保存成功的重要机制之一。

关键词(KeyWords)： 毛葡萄;缓慢生长法;生理响应;结构变化;种质保存

Abstract:

Keywords:

基金项目(Foundation): 贵州省优秀青年科技人才培养对象专项资金(黔科合人字[2009]02号)

作者(Authors): 潘学军;李霞;张文娥;

DOI: 10.13925/j.cnki.gsxb.2013.04.025

参考文献(References)：

• [1]ENGELMANN F.In vitro germplasm conservation[J].Acta Horticul-turae,1998(46):141-147.
• [2]ENGELMANN F.Use of biotechnologies for the conservation of plantbiodiversity[J].In Vitro Cellular and Development Biology Plant,2011,47:5-16.
• [3]IUCN.IUCN Red List of Endangered Species[N].In:http://www.iuc-nredlist.org/.2010.
• [4]BARBARA M R,VISWAMBHARAN S,MICHAEL K,ERIC B,VALERIE C P.Biodiversity conservation and conservation biotech-nology tools[J].In Vitro Cellular and Development Biology Plant,2011,47:1-4.
• [5]LU Xin-xiong,CHEN Xiao-ling.Progress of conservation and re-search of crop germplasm resources in China[J].Scientia AgriculturaSinica,2003,36(10):1125-1132.卢新雄,陈晓玲.我国作物种质资源保存与研究进展[J].中国农业科学,2003,36(10):1125-1132.
• [6]HENSHAW G G.Technical aspects of tissue culture storage for ge-netic conservation[C].In:Crop genetic resources for today and tomor-row.Cambridge University Press.Cambridge,Chap,1975:27.
• [7]MOREL G.Meristem culture techniques for the long term storage ofcultivated plants[C].In:Crop genetic resources for today and tomor-row.Cambridge University Press,Cambridge,Chap,1975:327-332.
• [8]MA Feng-wang,LI Jia-rui.Cryopreservation of apricot protoplasts[J].Acta Horticulturae Sinica,1998,25(4):329-332.马锋旺,李嘉瑞.杏原生质体的超低温保存[J].园艺学报,1998,25(4):329-332.
• [9]ZHAO Yan-hua,WU Yong-jie,ZHOU Ming-de.Cryopreservationof in vitro culture shoot tips of Prunus mahaleb[J].Acta HorticulturaeSinica,1999,26(6):402-403.赵艳华,吴永杰,周明德.马哈利樱桃离体超低温保存的研究[J].园艺学报,1999,26(6):402-403.
• [10]ZHAO Yan-hua,WU Yong-jie.Study on cryopreservation of in vitrocultured cabernet Franch 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.
• [11]WANG Zi-cheng,DENG Xiu-xin.Cryopreservation 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橘茎尖及植株再生[J].园艺学报,2001,28(4):301-306.
• [12]AI Peng-fei,LUO Zheng-rong.Conservation of in vitro shoots ofpersimmon and date plum by slow growth and genetic stability of re-covered plantlets[J].Acta Horticulturae Sinica,2004,31(4):441-446.艾鹏飞,罗正荣.柿和君迁子试管苗缓慢生长法保存及遗传稳定性研究[J].园艺学报,2004,31(4):441-446.
• [13]PENG Min-zhang,DENG Jiu-sheng,GAN Lin.Preservation ofanocado in vitro under low temperature[J].Journal of Fruit Science,1996,13(2):96-98.彭民璋,邓九生,甘霖.油梨种质的离体低温保存[J].果树科学,1996,13(2):96-98.
• [14]LI Chao-zhou,LI Wei,CAO Zi-yi.The role of S3307 in the growthand preservation of grape tube seedlings[J].Plant Physiology Com-munications,1997,33(1):18-20.李朝周,李唯,曹孜义.S3307在葡萄试管苗生长和保存中的作用(简报)[J].植物生理学通讯,1997,33(1):18-20.
• [15]LIU Yue-xue,LIU Xiao-jun,WANG Jia-fu,LIN Shun-quan.Ef-fects of low temperature and another condition on Eriobotryo japoni-ca germplasm preservation in vitro[J].Journal of Plant Resources andEnvironment,2004,13(1):28-31.刘月学,刘小军,王家福,林顺权.低温等因素对枇杷种质离体保存的影响[J].植物资源与环境学报,2004,13(1):28-31.
• [16]NIU Ai-guo,ZHANG Kai-chun,ZHANG Xiao-ming,YAN Guo-hua,LI Wen-sheng,JIANG Li-jie.Preservation of in vitro growthshoot of cherry germplasm[J].Acta Horticulturae Sinica,2005,32(2):298-300.牛爱国,张开春,张晓明,闫国华,李文生,姜立杰.樱桃种质资源试管苗保存方法[J].园艺学报,2005,32(2):298-300.
• [17]TEHRIM S,SAJID G M.In vitro establishment,conservation and itsimplications for grape germplasm biodiversity[J].Romanian Biotech-nological Letters,2011,16(6):6781-6789.
• [18]ZHU Wen-tao,ZHOU Hou-cheng,WANG Zi-cheng.Cryopreserva-tion of Fragaria pentapylla and analysis of genetic stability[J].Journalof Fruit Science,2013,30(1):55-61.朱文涛,周厚成,王子成.五叶草莓超低温保存及遗传稳定性分析[J].果树学报,2013,30(1):55-61.
• [19]ARNON D I.Copper enzymes in isolated chloroplasts.Polyphenolox-idase in Beta vulgaris[J].Plant Physiology,1949,24:1-15.
• [20]GAO Jun-feng.Plant physiology laboratory technology[M].Xi’an:World Publishing Corporation,2000.高俊凤.植物生理学实验技术[M].西安:世界图书出版社公司,2000.
• [21]GOPAL J,CHAUHAN N S.Slow growth in vitro conservation ofpotato germplasm at low temperature[J].Potato Research,2010,53:141-149.
• [22]LATA H,MORAES R M,BERTONI B,PEREIRA A M S.In vitrogermplasm conservation of Podophyllum peltatum L.under slowgrowth conditions[J].In vitro and Cellular and Development Biology-Plant,2010,46:22-27.
• [23]NEGASH A,KRENS F,SCHAART J,VISSER B.In vitro conser-vation of enset under slow-growth conditions[J].Plant Cell Tissue andOrgan Culture,2001,66:107-111.
• [24]SCHERWINSKI-PEREIRA J E,COSTA F H S,CAMILLO J,SIL-VA D B,ALVES R B N,VIEIRA R F.Tissue culture storage ofBrazilian medicinal plants germplasm[J].Acta Horticulturae,2010(860):211-214.
• [25]HU Jia-shu,ZHU Cheng,ZENG Guang-wen,ZHENG Guang-hua.Effect of aging-resistance of ultra-dried safflower seeds and itsmechanism[J].Acta Phytophysiologica Sinica,1999,25(2):171-177.胡家恕,朱诚,曾广文,郑光华.超干红花种子抗老化作用及其机理[J].植物生理学报,1999,25(2):171-177.
• [26]ZHU Cheng,ZENG Guang-wen,ZHENG Guang-hua.The storagetolerance and lipid peroxidation in ultradried peanut seeds[J].ActaAgronomica Sinica,2000,26(2):235-238.朱诚,曾广文,郑光华.超干花生种子耐藏性与脂质过氧化作用[J].作物学报,2000,26(2):235-238.