牛角瓜种子萌发和组培快繁技术研究
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摘要
【目的】对牛角瓜种子萌发及组培快繁技术进行研究,使其在短期内大量萌发和快速扩繁。【方法】用机械破壳处理使种子快速萌发,以无菌苗带芽茎段为外植体,选择不同的基本培养基和植物生长调节剂,对牛角瓜组培快繁技术进行研究。【结果】牛角瓜种子物理休眠使种子萌发不均一,机械破壳处理后种子萌发率达98%,发芽指数为29.6;与对照组相比,萌发率提高24%,发芽指数提高21。在MS+2.0 mg/L 6-BA培养基上培养时,外植体芽的诱导率最高及平均芽最多,分别为100%和4.9;最适生根培养基为WPM+0.15 mg/L IBA,其生根率和平均根数分别为100%和11.6条。以WPM为基本培养基的牛角瓜苗移栽成活率达90%,与以1/2 MS为基本培养基的牛角瓜相比,其成活率提高了30%。【结论】该研究提供了合适的牛角瓜种子萌发方法并建立了组培快繁体系,为其短期内大量繁殖提供理论基础与技术指导。
[Purpose]This study focused on the germination and tissue culture of Calotropis gigantea and aims to make it germination and propagation rapidly. [Method]The seeds were quickly germinated in mechanical treatment, and rapid propagation technology was studied by taking stem with buds of aseptic seedling as explants, and different basic media and plant growth regulators were tested. [Result]The seed germination of C. gigantea was not homogeneous because of the physical dormancy. The germination rate was 98% and the germination index was 29.6 after mechanical treatment. Compared with control group, their germination rate and index of the test material increased by 24% and 21, respectively. We found the ideal rapid propagation medium for explant bud was MS+2.0 mg/L 6-BA, and the average number of induced buds was 4.9 and the frequency of shoot induction was 100%. Whereas the optimal medium for root induction was WPM+0.15 mg/L IBA, and root frequency was up to 100%, and every explant had 11.6 roots. The survival rate of rooted saplings was up to 90% when using WPM as the basic culture medium. The survival rate increased by 30%compared with the rooted saplings which used 1/2 MS as basic culture medium. [Conclusion]This study identified the suitable seed germination method and the establishment of the rapid propagation system, which in turn provided the theoretical basis and technical guidance for the large propagation of the plant material in the short term.
[Purpose]This study focused on the germination and tissue culture of Calotropis gigantea and aims to make it germination and propagation rapidly. [Method]The seeds were quickly germinated in mechanical treatment, and rapid propagation technology was studied by taking stem with buds of aseptic seedling as explants, and different basic media and plant growth regulators were tested. [Result]The seed germination of C. gigantea was not homogeneous because of the physical dormancy. The germination rate was 98% and the germination index was 29.6 after mechanical treatment. Compared with control group, their germination rate and index of the test material increased by 24% and 21, respectively. We found the ideal rapid propagation medium for explant bud was MS+2.0 mg/L 6-BA, and the average number of induced buds was 4.9 and the frequency of shoot induction was 100%. Whereas the optimal medium for root induction was WPM+0.15 mg/L IBA, and root frequency was up to 100%, and every explant had 11.6 roots. The survival rate of rooted saplings was up to 90% when using WPM as the basic culture medium. The survival rate increased by 30%compared with the rooted saplings which used 1/2 MS as basic culture medium. [Conclusion]This study identified the suitable seed germination method and the establishment of the rapid propagation system, which in turn provided the theoretical basis and technical guidance for the large propagation of the plant material in the short term.
引文
[1]中国科学院植物志编辑委员会.中国植物志[M].北京:科学出版社,1977.
[2]ASHORI A,BAHREINI Z.Evaluation of Calotropis gigantea as a promising raw material for fiber-reinforced composite[J].Journal of Composite Materials,2009,43(11):1297.DOI:10.1177/0021998308104526.
[3]田湘,严理,唐丹,等.“全能灌木”牛角瓜研究热点及发展趋势[J].山西农业科学,2015,43(8):1061.DOI:10.3969/j.issn.1002-2481.2015.08.37.
[4]ZHENG Y A,ZHU Y F,WANG A Q,et al.Potential of Calotropis gigantea fiber as an absorbent for removal of oil from water[J].Industrial Crops and Products,2016,83:387.DOI:10.1016/j.indcrop.2016.01.009.
[5]ZHENG Y,CAO E J,TU L X,et al.A comparative study for oil-absorbing performance of octadecyltrichlorosilane treated Calotropis gigantea fiber and kapok fiber[J].Cellulose,2017,24(2):989.DOI:10.1007/s10570-016-1155-z.
[6]MBHELE N,BALOGUN F O,KAZEEM M I,et al.In vitro studies on the antimicrobial,antioxidant and antidiabetic potential of Cephalaria gigantea[J].Bangladesh Journal of Pharmacology,2015,10(1):214.DOI:10.3329/bjp.v10i1.21716.
[7]ADAK M,GUPTA J K.Evaluation of anti-inflammatory activity of Calotropis gigantea(AKANDA)in various biological system[J].Nepal Medical College Journal,2006,8(3):156.
[8]KOVENDAN K,MURUGAN K,KUMAR K P,et al.Mosquitocidal properties of Calotropis gigantea(Family:Asclepiadaceae)leaf extract and bacterial insecticide,Bacillus thuringiensis,against the mosquito vectors[J].Parasitology Research,2012,111(2):531.DOI:10.1007/s00436-012-2865-2.
[9]PATHAK A K,ARGAL A.Analgesic activity of Calotropis gigantea flower[J].Fitoterapia,2007,78(1):40.DOI:10.1016/j.fitote.2006.09.023.
[10]PARHIRA S,ZHU G Y,CHEN M,et al.Cardenolides from Calotropis gigantea as potent inhibitors of hypoxiainducible factor-1 transcriptional activity[J].Journal of Ethnopharmacology,2016,194(24):930.DOI:10.1016/j.jep.2016.10.070.
[11]CHITME H R,CHANDRA R,KAUSHIK S.Evaluation of antipyretic activity of Calotropis gigantea(Asclepiadaceae)in experimental animals[J].Phytotherapy Research,2005,19(5):454.DOI:10.1002/ptr.1672.
[12]CHITME H R,CHANDRA R,KAUSHIK S.Studies on anti-diarrheal activity of Calotropis gigantea R.Br.in experimental animals[J].Journal of Pharmacy and Pharmaceutical Sciences,2004,7(1):70.
[13]RATHOD N R,CHITME H R,IRCHHAIYA R,et al.Hypoglycemic effect of Calotropis gigantea Linn.leaves and flowers in streptozotocin-induced diabetic rats[J].Oman Medical Journal,2011,26(2):104.DOI:10.5001/omj.2011.26.
[14]PHOO ZWMM,RAZON LF,KNOTHE G,et al.Evaluation of Indian milkweed(Calotropis gigantea)seed oil as alternative feedstock for biodiesel[J].Industrial Crops and Products,2014,54(4):226.DOI:10.1016/j.indcrop.2014.01.029.
[15]魏静,赵元藩,张燕平.牛角瓜的栽培管理技术及应用前景[J].林业调查规划,2013,38(2):112.DOI:10.3969/j.issn.1671-3168.2013.02.026.
[16]ROY A T,KOUTOULIS A,DE D N.Cell suspension culture and plant regeneration in the latex-producing plant,Calotropis gigantea(Linn.)R.Br.[J].Plant Cell,Tissue and Organ Culture,2000,63(1):15.DOI:10.1023/A:1006497220647.
[17]ROY A T,DE D N.Tissue culture and plant regeneration from immature embryo explants of Calotropis gigantea(Linn.)R.Br.[J].Plant Cell,Tissue and Organ Culture,1990,20(3):229.
[18]李克烈,罗联忠,陈伟,等.牛角瓜的组织培养[J].广西农业生物科学,2007,26(3):247.
[19]孙健.牛角瓜及其发根中碳氢化合物与强心苷的代谢调控[D].北京:中国科学院,2012.
[20]唐军荣,李斌,刘惠民,等.牛角瓜离体快繁技术研究[J].云南农业大学学报(自然科学),2016,31(4):658.DOI:10.16211/j.issn.1004-390X(n).2016.04.012.
[21]刘进平,莫绕.热带植物组织培养[M].北京:科学出版社,2006.
[22]曹昆,李霞.木本植物组织培养不定芽诱导研究进展[J].江苏林业科技,2008,35(5):43.
[23]BEWLEY J D,BLACK M.Physiology and biochemistry of seeds in relation to germination[M].New York:Springer-Verlag,1978.
[24]LI P,WU H,GENG S L,et al.Germination and dormancy of seeds in Echinacea purpurea(L.)Moench(Asteraceae)[J].Seed Science and Technology,2007,35(1):9.DOI:10.15258/sst.2007.35.1.02.
[25]LI X J,BASKIN J M,BASKIN C C.Comparative morphology and physiology of fruit and seed development in the two shrubs Rhus aromatica and R.glabra(Anacardiaceae)[J].Merican Journal of Botany,1999,86(9):1217.DOI:10.2307/2656769.
[26]JAYASURIYA K M G G,BASKIN J M,GENEVE R L,et al.Seed development in Ipomoea lacunosa(Convolvulaceae),with particular reference to anatomy of the water gap[J].Annals of Botany,2007,100(3):459.DOI:10.1093/aob/mcm137.
[27]罗瑛,吴开永,刘国道.不同处理对银合欢种子发芽率的影响[J].安徽农业科学,2009,37(13):6227.DOI:10.13989/j.cnki.0517-6611.2009.13.113.
[28]连洁琼.三种豆科树种硬实形成机制及破除方法的初步研究[D].泰安:山东农业大学,2013.
[29]El-KEBLAWY A.Germination response to light and temperature in eight annual grasses from disturbed and natural habitats of an arid Arabian desert[J].Journal of Arid Environments,2017,147:1.DOI:10.1016/j.jaridenv.2017.08.002.
[30]程世昌,王小琳.重庆植物组织培养[M].重庆:重庆大学出版社,2011.
[31]马燕,韩瑞超,臧德奎,等.木本观赏植物组织培养研究进展[J].安徽农业科学,2012,40(4):1956.DOI:10.13989/j.cnki.0517-6611.2012.04.201.
[32]徐晓峰,黄学林.TDZ:一种有效的植物生长调节剂[J].植物学通报,2003,20(2):227.
[33]沈烨.根癌农杆菌介导的甜樱桃茎尖遗传转化体系的建立[D].重庆:西南农业大学,2002.
[34]施翔.杂交松组织培养研究[D].南京:南京林业大学,2005.
[35]KUMAWAT S,KACHHWAHA S,KOTHARI S L.Micronutrient optimization in the basal medium improves in vitro plant regeneration in carnation(Dianthus caryophyllus L.)and Chinese pink(Dianthus chinensis L.)[J].Propagation of Ornamental Plants,2013,13(1):3.
[36]HAND C,REED B M.Minor nutrients are critical for the improved growth of Corylus avellana shoot cultures[J].Plant Cell Tissue and Organ Culture,2014,119(2):427.DOI:10.1007/s11240-014-0545-x.
[37]郭颖.三种高山杜鹃组织培养快繁技术研究[D].北京:北京林业大学,2015.
[38]程雪梅.马缨杜鹃种子萌发与组织培养技术体系研究[D].昆明:西南林学院,2008.
[39]刘会超,贾文庆.基本培养基及植物生长调节剂对牡丹组培苗生根的影响[J].河南科技学院学报(自然科学版),2010,38(2):32.DOI:10.3969/j.issn.1008-7516.2010.02.008.
[2]ASHORI A,BAHREINI Z.Evaluation of Calotropis gigantea as a promising raw material for fiber-reinforced composite[J].Journal of Composite Materials,2009,43(11):1297.DOI:10.1177/0021998308104526.
[3]田湘,严理,唐丹,等.“全能灌木”牛角瓜研究热点及发展趋势[J].山西农业科学,2015,43(8):1061.DOI:10.3969/j.issn.1002-2481.2015.08.37.
[4]ZHENG Y A,ZHU Y F,WANG A Q,et al.Potential of Calotropis gigantea fiber as an absorbent for removal of oil from water[J].Industrial Crops and Products,2016,83:387.DOI:10.1016/j.indcrop.2016.01.009.
[5]ZHENG Y,CAO E J,TU L X,et al.A comparative study for oil-absorbing performance of octadecyltrichlorosilane treated Calotropis gigantea fiber and kapok fiber[J].Cellulose,2017,24(2):989.DOI:10.1007/s10570-016-1155-z.
[6]MBHELE N,BALOGUN F O,KAZEEM M I,et al.In vitro studies on the antimicrobial,antioxidant and antidiabetic potential of Cephalaria gigantea[J].Bangladesh Journal of Pharmacology,2015,10(1):214.DOI:10.3329/bjp.v10i1.21716.
[7]ADAK M,GUPTA J K.Evaluation of anti-inflammatory activity of Calotropis gigantea(AKANDA)in various biological system[J].Nepal Medical College Journal,2006,8(3):156.
[8]KOVENDAN K,MURUGAN K,KUMAR K P,et al.Mosquitocidal properties of Calotropis gigantea(Family:Asclepiadaceae)leaf extract and bacterial insecticide,Bacillus thuringiensis,against the mosquito vectors[J].Parasitology Research,2012,111(2):531.DOI:10.1007/s00436-012-2865-2.
[9]PATHAK A K,ARGAL A.Analgesic activity of Calotropis gigantea flower[J].Fitoterapia,2007,78(1):40.DOI:10.1016/j.fitote.2006.09.023.
[10]PARHIRA S,ZHU G Y,CHEN M,et al.Cardenolides from Calotropis gigantea as potent inhibitors of hypoxiainducible factor-1 transcriptional activity[J].Journal of Ethnopharmacology,2016,194(24):930.DOI:10.1016/j.jep.2016.10.070.
[11]CHITME H R,CHANDRA R,KAUSHIK S.Evaluation of antipyretic activity of Calotropis gigantea(Asclepiadaceae)in experimental animals[J].Phytotherapy Research,2005,19(5):454.DOI:10.1002/ptr.1672.
[12]CHITME H R,CHANDRA R,KAUSHIK S.Studies on anti-diarrheal activity of Calotropis gigantea R.Br.in experimental animals[J].Journal of Pharmacy and Pharmaceutical Sciences,2004,7(1):70.
[13]RATHOD N R,CHITME H R,IRCHHAIYA R,et al.Hypoglycemic effect of Calotropis gigantea Linn.leaves and flowers in streptozotocin-induced diabetic rats[J].Oman Medical Journal,2011,26(2):104.DOI:10.5001/omj.2011.26.
[14]PHOO ZWMM,RAZON LF,KNOTHE G,et al.Evaluation of Indian milkweed(Calotropis gigantea)seed oil as alternative feedstock for biodiesel[J].Industrial Crops and Products,2014,54(4):226.DOI:10.1016/j.indcrop.2014.01.029.
[15]魏静,赵元藩,张燕平.牛角瓜的栽培管理技术及应用前景[J].林业调查规划,2013,38(2):112.DOI:10.3969/j.issn.1671-3168.2013.02.026.
[16]ROY A T,KOUTOULIS A,DE D N.Cell suspension culture and plant regeneration in the latex-producing plant,Calotropis gigantea(Linn.)R.Br.[J].Plant Cell,Tissue and Organ Culture,2000,63(1):15.DOI:10.1023/A:1006497220647.
[17]ROY A T,DE D N.Tissue culture and plant regeneration from immature embryo explants of Calotropis gigantea(Linn.)R.Br.[J].Plant Cell,Tissue and Organ Culture,1990,20(3):229.
[18]李克烈,罗联忠,陈伟,等.牛角瓜的组织培养[J].广西农业生物科学,2007,26(3):247.
[19]孙健.牛角瓜及其发根中碳氢化合物与强心苷的代谢调控[D].北京:中国科学院,2012.
[20]唐军荣,李斌,刘惠民,等.牛角瓜离体快繁技术研究[J].云南农业大学学报(自然科学),2016,31(4):658.DOI:10.16211/j.issn.1004-390X(n).2016.04.012.
[21]刘进平,莫绕.热带植物组织培养[M].北京:科学出版社,2006.
[22]曹昆,李霞.木本植物组织培养不定芽诱导研究进展[J].江苏林业科技,2008,35(5):43.
[23]BEWLEY J D,BLACK M.Physiology and biochemistry of seeds in relation to germination[M].New York:Springer-Verlag,1978.
[24]LI P,WU H,GENG S L,et al.Germination and dormancy of seeds in Echinacea purpurea(L.)Moench(Asteraceae)[J].Seed Science and Technology,2007,35(1):9.DOI:10.15258/sst.2007.35.1.02.
[25]LI X J,BASKIN J M,BASKIN C C.Comparative morphology and physiology of fruit and seed development in the two shrubs Rhus aromatica and R.glabra(Anacardiaceae)[J].Merican Journal of Botany,1999,86(9):1217.DOI:10.2307/2656769.
[26]JAYASURIYA K M G G,BASKIN J M,GENEVE R L,et al.Seed development in Ipomoea lacunosa(Convolvulaceae),with particular reference to anatomy of the water gap[J].Annals of Botany,2007,100(3):459.DOI:10.1093/aob/mcm137.
[27]罗瑛,吴开永,刘国道.不同处理对银合欢种子发芽率的影响[J].安徽农业科学,2009,37(13):6227.DOI:10.13989/j.cnki.0517-6611.2009.13.113.
[28]连洁琼.三种豆科树种硬实形成机制及破除方法的初步研究[D].泰安:山东农业大学,2013.
[29]El-KEBLAWY A.Germination response to light and temperature in eight annual grasses from disturbed and natural habitats of an arid Arabian desert[J].Journal of Arid Environments,2017,147:1.DOI:10.1016/j.jaridenv.2017.08.002.
[30]程世昌,王小琳.重庆植物组织培养[M].重庆:重庆大学出版社,2011.
[31]马燕,韩瑞超,臧德奎,等.木本观赏植物组织培养研究进展[J].安徽农业科学,2012,40(4):1956.DOI:10.13989/j.cnki.0517-6611.2012.04.201.
[32]徐晓峰,黄学林.TDZ:一种有效的植物生长调节剂[J].植物学通报,2003,20(2):227.
[33]沈烨.根癌农杆菌介导的甜樱桃茎尖遗传转化体系的建立[D].重庆:西南农业大学,2002.
[34]施翔.杂交松组织培养研究[D].南京:南京林业大学,2005.
[35]KUMAWAT S,KACHHWAHA S,KOTHARI S L.Micronutrient optimization in the basal medium improves in vitro plant regeneration in carnation(Dianthus caryophyllus L.)and Chinese pink(Dianthus chinensis L.)[J].Propagation of Ornamental Plants,2013,13(1):3.
[36]HAND C,REED B M.Minor nutrients are critical for the improved growth of Corylus avellana shoot cultures[J].Plant Cell Tissue and Organ Culture,2014,119(2):427.DOI:10.1007/s11240-014-0545-x.
[37]郭颖.三种高山杜鹃组织培养快繁技术研究[D].北京:北京林业大学,2015.
[38]程雪梅.马缨杜鹃种子萌发与组织培养技术体系研究[D].昆明:西南林学院,2008.
[39]刘会超,贾文庆.基本培养基及植物生长调节剂对牡丹组培苗生根的影响[J].河南科技学院学报(自然科学版),2010,38(2):32.DOI:10.3969/j.issn.1008-7516.2010.02.008.