大苞白山茶胚性愈伤的诱导及植株再生
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摘要
大苞白山茶(Camellia granthamiana)是我国特有珍稀木本植物,具有重要的理论研究和应用价值。本研究以幼胚子叶为外植体,探究不同植物生长调节剂配比对愈伤形成、体胚发生及体胚诱导成苗的影响。结果表明, 2.0 mg·L~(-1) 6-BA与2.5 mg·L~(-1) 2,4-D协同作用下,愈伤组织诱导效果最好,诱导率最高可达100%,但形成的均为非胚性愈伤; TDZ浓度为0.05 mg·L~(-1)时,胚性愈伤诱导率可达85.47%;体胚诱导的最佳培养基为MS+0.5mg·L~(-1) 6-BA+0.05 mg·L~(-1) NAA+0.05 mg·L~(-1) TDZ,每克胚性愈伤体胚数可达162个,其正常体胚率达58.34%;在1/2MS+1.5 mg·L~(-1) 6-BA+0.05 mg·L~(-1) NAA中,体胚成苗率最高,达33.91%;试管苗移栽于熟化红壤土:木屑:有机肥=5:4:1的基质中,成活率可达80%以上。
Camellia granthamiana is a rare and endemic woody plant in China with vital theoretical research and application value. Using young embryo cotyledon as explants, we investigated the effects of different plant growth regulators on the induction of callus, somatic embryogenesis, and seedling rate of Camellia granthamiana. The results showed that the callus induction rate was up to 100% under the combination of 2.0 mg·L~(-1)6-BA with 2.5 mg·L~(-1) 2,4-D, however, all of them were none-embryonic callus. When TDZ concentration was0.05 mg·L~(-1), the embryonic callus induction rate was 85.47%. The best medium for the induction of somatic embryogenesis was MS+0.5 mg·L~(-1) 6-BA+0.05 mg·L~(-1) NAA+0.05 mg·L~(-1) TDZ and the number of embryonic callus embryo was 162 per gram, and normal somatic embryo rate was 58.34%. In the medium with 1/2 MS+1.5 mg·L~(-1) 6-BA+0.05 mg·L~(-1) NAA, the rate of embryo formation was the highest that reached 33.91%. More than80% of the plantlets survived after transplanted in substrate with red soil: wood chips: organic fertilizer=5:4:1.
Camellia granthamiana is a rare and endemic woody plant in China with vital theoretical research and application value. Using young embryo cotyledon as explants, we investigated the effects of different plant growth regulators on the induction of callus, somatic embryogenesis, and seedling rate of Camellia granthamiana. The results showed that the callus induction rate was up to 100% under the combination of 2.0 mg·L~(-1)6-BA with 2.5 mg·L~(-1) 2,4-D, however, all of them were none-embryonic callus. When TDZ concentration was0.05 mg·L~(-1), the embryonic callus induction rate was 85.47%. The best medium for the induction of somatic embryogenesis was MS+0.5 mg·L~(-1) 6-BA+0.05 mg·L~(-1) NAA+0.05 mg·L~(-1) TDZ and the number of embryonic callus embryo was 162 per gram, and normal somatic embryo rate was 58.34%. In the medium with 1/2 MS+1.5 mg·L~(-1) 6-BA+0.05 mg·L~(-1) NAA, the rate of embryo formation was the highest that reached 33.91%. More than80% of the plantlets survived after transplanted in substrate with red soil: wood chips: organic fertilizer=5:4:1.
引文
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Wu B,Li JY,Fan ZQ,et al(2013).Callus induction and plant regeneration of Camellia japonica L.‘Naidong’.Plant Physiol J,49:343-346(in Chinese with English abstract)[吴斌,李纪元,范正琪等(2013).山茶‘耐冬’愈伤组织诱导与植株再生.植物生理学报,49:343-346]
Yang J,Wu S,Li C(2013).High efficiency secondary somatic embryogenesis in Hovenia dulcis Thunb.through solid and liquid cultures.Sci World J,2013:718754
Yang JF,Liu JM,Huang YX,et al(2013).Effects of different plant growth regulators on callus induction from leaf explant of Camellia granthamina.Guangdong Agric Sci,40:152-154(in Chinese with English abstract)[杨建芬,刘金梅,黄云香等(2013).不同植物生长调节剂对大苞白山茶叶片愈伤组织诱导的影响.广东农业科学,40:152-154]
Yang K(2013).Induction of somatic embryogenesis and embryogenic callus from Camellia japonica.J Qingdao Agric Univ(Nat Sci),30:184-187(in Chinese with English abstract)[杨凯(2013).耐冬山茶胚状体和胚性愈伤组织诱导研究.青岛农业大学学报(自然科学版),30:184-187]
Yang X,Zhang X(2010).Regulation of somatic embryogenesis in higher plants.Crit Rev Plant Sci,29:36-57
Ge LW,Guo W,Pan ZK,et al(2015).The research literature analysis about tissue culture of genus Camellia in China.J Fujian Forestry Sci Tech,42:237-241(in Chinese with English abstract)[葛立雯,郭维,潘正康等(2015).我国山茶属植物组织培养研究文献分析.福建林业科技,42:237-241]
Isah T(2016).Induction of somatic embryogenesis in woody plants.Acta Physiol Plant,38:118
Jayasankar S,Bondada BR,Li Z,et al(2003).Comparative anatomy and morphology of Vitis vinifera(Vitaceae)somatic embryos from solid and liquid culture systems.Am J Bot,90:973-979
LüJF,Chen R,Zhang MH,et al(2013).Plant regeneration via somatic embryogenesis and shoot organogenesis from immature cotyledons of Camellia nitidissima Chi.J Plant Physiol,170:1202-1211
Min TL,Bartholomew B(2007).Theaceae.In:Wu ZY,Raven PH(eds).Flora of China.Beijing:Science Press,12:366-478
Qin HN,Yang Y,Dong SY,et al(2017).Threatened species list of China’s higher plants.Biodivers Sci,25:696-744[覃海宁,杨永,董仕勇等(2017).中国高等植物受威胁物种名录.生物多样性,25:696-744]
Roy PK,Roy SK,Hakim L,et al(2016).Somatic embryogenesis and plant regeneration of papaya(Carica papaya L.cv.Shahi).Nucl Sci Appl,25:23-26
Shahzad A,Parveen S,Sharma S,et al(2017).Plant Tissue Culture:Applications in Plant Improvement and Conservation.In:Abdin M,Kiran U,Kamaluddin,et al(eds).Plant Biotechnology:Principles and Applications.Singapore:Springer,37-72
Tong HY,Zhou XX,Li J,et al(2018).Tissue culture and rapid propagation of rare and endangered Tangtsinia nanchuanica.Plant Physiol J,54:1681-1686[童虹宇,周小雪,李娟等(2018).珍稀濒危植物金佛山兰的组培快繁.植物生理学报,54:1681-1686]
Wu B,Li JY,Fan ZQ,et al(2013).Callus induction and plant regeneration of Camellia japonica L.‘Naidong’.Plant Physiol J,49:343-346(in Chinese with English abstract)[吴斌,李纪元,范正琪等(2013).山茶‘耐冬’愈伤组织诱导与植株再生.植物生理学报,49:343-346]
Yang J,Wu S,Li C(2013).High efficiency secondary somatic embryogenesis in Hovenia dulcis Thunb.through solid and liquid cultures.Sci World J,2013:718754
Yang JF,Liu JM,Huang YX,et al(2013).Effects of different plant growth regulators on callus induction from leaf explant of Camellia granthamina.Guangdong Agric Sci,40:152-154(in Chinese with English abstract)[杨建芬,刘金梅,黄云香等(2013).不同植物生长调节剂对大苞白山茶叶片愈伤组织诱导的影响.广东农业科学,40:152-154]
Yang K(2013).Induction of somatic embryogenesis and embryogenic callus from Camellia japonica.J Qingdao Agric Univ(Nat Sci),30:184-187(in Chinese with English abstract)[杨凯(2013).耐冬山茶胚状体和胚性愈伤组织诱导研究.青岛农业大学学报(自然科学版),30:184-187]
Yang X,Zhang X(2010).Regulation of somatic embryogenesis in higher plants.Crit Rev Plant Sci,29:36-57