麻疯树根高效再生体系的建立及不定芽起源研究
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摘要
为了克服现有麻疯树快繁技术的不足,分别以麻疯树胚根和子叶外植体诱导的不定根为外植体诱导不定芽.麻疯树子叶不定根诱导的最佳培养方式为:MS+5 mg/L IBA上培养4天,然后转至MS0培养基上生根.诱导生根率和生根数分别达到93.33%和6.54个/外植体.子叶不定根诱导不定芽的最佳培养基是MS+3.0 mg/L 6-BA+3 mg/L IAA,诱导率达到83.33%.麻疯树胚根的最佳不定芽诱导培养基为MS+0.2 mg/L 6-BA+1.0 mg/L IBA,分化率达67.80%,平均出芽数为3.25个/外植体.对麻疯树胚根再生的各阶段进行了石蜡切片观察,明确了不定芽起源于根的薄壁细胞层.与现有技术相比,本方法具有周期短、再生效率高的特点.
In order to overcome the shortcomings of the existing propagation technology of Jatropha curcas, adventitious buds were induced from the adventitious root explants of cotyledon and radicle explants respectively. The optimal way to get adventitious root from cotyledons of Jatropha curcas L. was that it would be precultured on the MS medium containing 5 mg/L IBA for 4 days, then transferred to MS_0 medium for rooting. The rooting inducing rate and adventitious root number reached 93.33% and 6.54/explant, respectively. The optimal medium for adventitious buds inducing was MS+3.0 mg/L 6-BA+3 mg/L IAA. The induction rate could reach 83.33%. The optimal adventitious bud induction medium from the radicle explants of Jatropha curcas L. was MS+0.2 mg/L 6-BA+1.0 mg/L IBA. The differentiation rate was 67.80%, and the average number of sprouts was 3.25 per explant. Paraffin sections were observed at each stage of bud regeneration and it seems that adventitious buds originate from parenchyma of roots. Compared with the prior methodology, the method presented here has the characteristics of a shorter cycle and higher regeneration efficiency.
In order to overcome the shortcomings of the existing propagation technology of Jatropha curcas, adventitious buds were induced from the adventitious root explants of cotyledon and radicle explants respectively. The optimal way to get adventitious root from cotyledons of Jatropha curcas L. was that it would be precultured on the MS medium containing 5 mg/L IBA for 4 days, then transferred to MS_0 medium for rooting. The rooting inducing rate and adventitious root number reached 93.33% and 6.54/explant, respectively. The optimal medium for adventitious buds inducing was MS+3.0 mg/L 6-BA+3 mg/L IAA. The induction rate could reach 83.33%. The optimal adventitious bud induction medium from the radicle explants of Jatropha curcas L. was MS+0.2 mg/L 6-BA+1.0 mg/L IBA. The differentiation rate was 67.80%, and the average number of sprouts was 3.25 per explant. Paraffin sections were observed at each stage of bud regeneration and it seems that adventitious buds originate from parenchyma of roots. Compared with the prior methodology, the method presented here has the characteristics of a shorter cycle and higher regeneration efficiency.
引文
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[14]Datta M M,Mukherjee P,Ghosh B,et al.In vitro clonal propagation of biodiesel plant(Jatropha curcas L.)[J].Curr Sci India,2007,93:1438.
[15]Kalimuthu K,Paulsamy S,Senthilkumar R,et al.In vitro propagation of the biodiesel plant Jatropha curcas L.[J].Plant Tiss Cult Biotech,2007,17:137.
[16]Kaewpoo M,Te-Chato S.Study on ploidy level of micropropagated Jatropha curcas L.via flow cytometry[J].Intern J Agri Tech,2010,6:391.
[17]Sharma S,Kumar N,Reddy M P.Regeneration in Jatropha curcas:Factors affecting the efficiency of in vitro regeneration[J].Ind Crops Prod,2011,34:943.
[18]Wei Q,Lu W D,Liao Y,et al.Plant regeneration from epicotyl explant of Jatropha curcas[J].JPlant Physio Mol Bio,2004,30:475.
[19]杨千,吉柔风,李晨阳,等.川滇琼地区麻疯树种子核糖体失活蛋白资源调查[J].四川大学学报:自然科学版,2017,54:197.
[20]刘伯斌,卢孟柱,李玲,等.麻疯树叶盘法高效再生的研究[J].林业科学研究,2010,23:326.
[21]穆燕,侯金艳,陈晶,等.油桐下胚轴直接再生体系的建立[J].分子植物育种,2016,4:1821.
[22]张艳玲,吴秀华,周月,等.‘秦美’猕猴桃叶片高频直接再生体系的建立[J].西南大学学报:自然科学版,2016,38:20.
[23]Kumar N,Reddy M P.Plant regeneration through the direct induction of shoot buds from petiole explants of Jatropha curcas:a biofuel plant[J].Ann Appl Bio,2010,156:367.
[24]Li C,Fu Q,Niu L,et al.Three TFL1homologues regulate floral initiation in the biofuel plant Jatropha curcas[J].Sci Rep-UK,2017,7:43090.
[2]Clark T E,Appleton C C,Kvalsvig J D.Schistosomiasis and the use of indigenous.plant molluscicides:a rural south African perspective[J].Acta Trop,1997,66:93.
[3]中国科学院中国植物志编辑委员会.中国植物志:第44卷第2分册[M].北京:科学出版社,1996.
[4]郑万钧.中国树木志(第3卷)[M].北京:中国林业出版社,1998.
[5]Deore A C,Johnson T S.High-frequency plant regeneration from leaf-disc cultures of Jatropha curcas L.:an important biodiesel plant[J].Plant Biotechnol Rep,2008,2:7.
[6]Khurana-Kaul V,Kachhwaha S,Kothari S L.Direct shoot regeneration from leaf explants of Jatropha curcas in response to thidiazuron and high copper contents in the medium[J].Biol Plantarum,2010,54:369.
[7]Zhang C,Fu S,Tang G,et al.Factors influencing direct shoot regeneration from mature leaves of Jatrophacurcas,an important biofuel plant[J].In Vitro Cell Dev Biol-Plant,2013,49:529.
[8]Medipally S R,Naresh B,Kumar S M,et al.Somatic embryogenesis from leaf and shoot tip explants of Jatropha curcas L.[J].Indian Sci Tech,2014,7:1842.
[9]王琼,刘伯斌,孔倩倩,等.能源植物麻疯树分子育种研究进展[J].生物技术通报,2014,3:1.
[10]Kumar N,Anand K G V,Reddy M P.Shoot regeneration from cotyledonary leaf explants of Jatropha curcas:a biodiesel plant[J].Acta Physiol Plant,2010,32:917.
[11]Kumar N,Reddy M P.Thidiazuron(TDZ)induced plant regeneration from cotyledonary petiole explants of elite genotypes of Jatropha curcas:a candidate biodiesel plant[J].Ind Crops Prod,2012,39:62.
[12]Kumar N,Anand K G V,Reddy M P.In vitro regeneration from petiole explants of non-toxic Jatropha curcas[J].Ind Crop Prod,2011,33:146.
[13]Rajore S,Batra A.Efficient plant regeneration via shoot tip explant in Jatropha curcas L.[J].J Plant Biochem Biotech,2005,14:73.
[14]Datta M M,Mukherjee P,Ghosh B,et al.In vitro clonal propagation of biodiesel plant(Jatropha curcas L.)[J].Curr Sci India,2007,93:1438.
[15]Kalimuthu K,Paulsamy S,Senthilkumar R,et al.In vitro propagation of the biodiesel plant Jatropha curcas L.[J].Plant Tiss Cult Biotech,2007,17:137.
[16]Kaewpoo M,Te-Chato S.Study on ploidy level of micropropagated Jatropha curcas L.via flow cytometry[J].Intern J Agri Tech,2010,6:391.
[17]Sharma S,Kumar N,Reddy M P.Regeneration in Jatropha curcas:Factors affecting the efficiency of in vitro regeneration[J].Ind Crops Prod,2011,34:943.
[18]Wei Q,Lu W D,Liao Y,et al.Plant regeneration from epicotyl explant of Jatropha curcas[J].JPlant Physio Mol Bio,2004,30:475.
[19]杨千,吉柔风,李晨阳,等.川滇琼地区麻疯树种子核糖体失活蛋白资源调查[J].四川大学学报:自然科学版,2017,54:197.
[20]刘伯斌,卢孟柱,李玲,等.麻疯树叶盘法高效再生的研究[J].林业科学研究,2010,23:326.
[21]穆燕,侯金艳,陈晶,等.油桐下胚轴直接再生体系的建立[J].分子植物育种,2016,4:1821.
[22]张艳玲,吴秀华,周月,等.‘秦美’猕猴桃叶片高频直接再生体系的建立[J].西南大学学报:自然科学版,2016,38:20.
[23]Kumar N,Reddy M P.Plant regeneration through the direct induction of shoot buds from petiole explants of Jatropha curcas:a biofuel plant[J].Ann Appl Bio,2010,156:367.
[24]Li C,Fu Q,Niu L,et al.Three TFL1homologues regulate floral initiation in the biofuel plant Jatropha curcas[J].Sci Rep-UK,2017,7:43090.