茎秆的第一节间离体再生空心菜
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摘要
以空心菜(Ipomoea aquatica Forsk.)品种‘白骨柳叶’为材料,通过筛选植物外植体和调整培养基激素配比等方法,首次建立了茎秆第一节间为外植体的空心菜离体再生体系。结果表明,在MS基本培养上添加0.05%的植物组织培养抗菌剂PPM可获得大量空心菜无菌苗;植株子叶和下胚轴的切段均未诱导出不定芽,而茎秆第一节间为外植体能成功诱导出不定芽,诱导成功率为20%,最佳培养基配方为MS+1.0 mg/L 6-BA+0.1 mg/L IAA;不定芽诱导生根的最佳培养基配方为1/2 MS+0.1 mg/L NAA,生根率为100%。诱导成功后,将完整的再生苗移栽至基质土中,成活率可达100%。
In this experiment,white bone willow leaf spinach(Ipomoea aquatica Forsk.) was used as research material.By screening explants and hormone ratios,an in vitro regeneration system of I.aquatica from first stem internode explants was established for the first time.Results showed that a large number of sterile I.aquatica seedlings could be obtained by adding 0.05% PPM to the basic culture of MS.Explants of cotyledons and hypocotyls did not induce adventitious buds;however,the first stem internode successfully induced adventitious buds.The optimal medium was MS+1.0 mg/L 6-BA+0.1 mg/L IAA,and the adventitious bud induction rate was 20%.The optimal medium for inducing adventitious bud roots was 1/2 MS +0.1 mg/L NAA,with the rooting rate reaching 100%.The whole regenerated seedlings were transplanted into matrix soil,with a transplant survival rate of 100%.This study established a spinach regeneration system and laid the foundation for further research on transgenic I.aquatica.
In this experiment,white bone willow leaf spinach(Ipomoea aquatica Forsk.) was used as research material.By screening explants and hormone ratios,an in vitro regeneration system of I.aquatica from first stem internode explants was established for the first time.Results showed that a large number of sterile I.aquatica seedlings could be obtained by adding 0.05% PPM to the basic culture of MS.Explants of cotyledons and hypocotyls did not induce adventitious buds;however,the first stem internode successfully induced adventitious buds.The optimal medium was MS+1.0 mg/L 6-BA+0.1 mg/L IAA,and the adventitious bud induction rate was 20%.The optimal medium for inducing adventitious bud roots was 1/2 MS +0.1 mg/L NAA,with the rooting rate reaching 100%.The whole regenerated seedlings were transplanted into matrix soil,with a transplant survival rate of 100%.This study established a spinach regeneration system and laid the foundation for further research on transgenic I.aquatica.
引文
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[31]王霞霞,马晖玲,曹冬梅.大花萱草‘香宝’再生体系的建立[J].甘肃农业大学学报,2014,49(4):136-142.Wang XX,Ma HL,Cao DM.Establishment of regeneration system of Hemeroallis middendorfii Trautv.et Mey‘Sweet Treasure’[J].Journal of Gansu Agricultural University,2014,49(4):136-142.
[32]王金祥,严小龙,潘瑞炽.不定根形成与植物激素的关系[J].植物生理学报,2005,41(2):133-142.Wang JX,Yan XL,Pan RZ.Relationship between adventitious root formation and plant hormones[J].Plant Physiology Communications,2005,41(2):133-142.
[33]席银凯,王元忠,黄衡宇,钱子刚.滇龙胆丛芽高效诱导与植株再生体系的建立[J].中草药,2018,49(6):1398-1404.Xi YK,Wang YZ,Huang HY,Qian ZG.Efficient induction system of adventitious shoot and plantlet regeneration in Gentiana rigescens[J].Chinese Traditional and Herbal Drugs,2018,49(6):1398-1404.
[34]Collado R,Veitía N,Bermúdez-Caraballoso I,García LR,Torres D,et al.Efficient in vitro plant regeneration via indirect organogenesis for different common bean cultivars[J].Sci Hortic,2013,153(3):109-116.
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[2]李法君.北方地区空心菜高效立体种植模式[J].北方园艺,2016,19(210):210.Li FJ.High-efficiency three-dimensional planting mode of water spinach in northern areas[J].Northern Horticulture,2016,19(210):210.
[3]朱宝鹏.日光温室夏秋季空心菜栽培技术[J].黑龙江农业科学,2008,(3):163.Zhu GP.Cultivation techniques of water spinach in solar greenhouse in summer and autumn[J].Heilongjiang Agricultural Sciences,2008,(3):163.
[4]宋喜军,朱德波,邓丙祥.北方栽培空心菜的关键技术[J].中国园艺文摘,2010,26(11):129.Song XJ,Zhu DB,Deng BX.Key techniques for cultivating water spinach in north China[J].Chinese Horticulture Abstracts,2010,26(11):129.
[5]Umar KJ,Hassan LG,Dangoggo SM,Ladan MJ.Nutritional composition of water spinach(Ipomoea aquatica Forsk.)leaves[J].J Appl Sci,2007,7(6):803-809.
[6]Malalavidhane TS,Wickramasinghe SM,Jansz ER.Oral hypoglycaemic activity of Ipomoea aquatica[J].J Ethnopharmacol,2000,72(1-2):293-298.
[7]Furukawa K.Advanced treatment and food production by hydroponic type wastewater treatment plant[J].Wat Sci Tech,2011,28(2):219-228.
[8]Liu QC,Zhai H,Wang Y,Zhang DP.Efficient plant regeneration from embryogenic suspension cultures of sweet potato[J].In Vitro Cell Dev-Pl,2001,37(5):564-567.
[9]Gonzalez RG,Sanchez DS,Campos JM,Vázquez EP,Guerra ZZ,et al.Plant regeneration from leaf and stem explants from two sweet potato(Ipomoea batatas(L.)Lam.)cultivars[J].Biotecnologia Aplicada,1999,16(1):11-14.
[10]张先云,李长看,袁秀云.甘薯高效再生体系的建立[J].安徽农业科学,2010,38(13):6666-6667.Zhang XY,Li ZK,Yuan XY.Establishment of efficient regeneration system of Ipomoea batatas[J].Journal of Anhui Agricultural Sciences,2010,38(13):6666-6667.
[11]张晓军,张永乐.日本牵牛茎段的离体培养及植株再生[J].植物生理学报,2004,40(6):717.Zhang XJ,Zhang YL.In vitro culture and plantlet regeneration of stem segment of Pharbitis nil[J].Plant Physiology Communications,2004,40(6):717.
[12]张希太,李俊玲,宋九英,宁书祥,陈海俊.巴西牵牛的组织培养技术研究[J].河南科技学院学报:自然科学版,2006,34(1):41-44.Zhang XT,Li JL,Song JY,Ning SX,Chen HJ.Studys on tissue culture of Ipomoea Setosa[J].Journal of Henan Institute of Science and Technology:Natural Science Edition,2006,34(1):41-44.
[13]Jia SR,Chua NH.Somatic embryogenesis and plant regeneration from immature embryo cultute of Pharbitis nil[J].Plant Sci,1992,87:215-223.
[14]黄兰.紫甘薯抗寒特性及组织培养研究[D].南昌:南昌大学,2012.
[15]薛启汉.甘薯子叶愈伤组织诱导与植株分化[J].江苏农业学报,1987,3(3):25-32.Xue QH.Callus induction and plantlet regeneration of sweet potatp cptyledon cultured in Vitro[J].Jiangsu Journal of Agricultural Sciences,1987,3(3):25-32.
[16]Gosukonda RM,Porobodessai A,Blay E,Prakash CS,Peterson CM.Thidiazuron-induced adventitious shoot regeneration of sweet potato(Ipomoea batatas)[J].In Vitro Cell Dev-Pl,1995,31(2):65-71.
[17]Sivparsad BJ,Gubba A.Development of an efficient plant regeneration protocol for sweet potato(Ipomoea batatas L.)cv.Blesbok[J].Afr J Biotech,2012,11(84):14982-14987.
[18]姜长阳,庞婧,刘宁,邹玉英,邹翠霞.空心菜无性系的建立及快速繁殖[J].辽宁大学学报:自然科学版,2004,31(3):275-278.Jiang CY,Pang J,Liu N,Zhou YY,Zhou CX.Establishment of regeneration clones and rapid propagation of Lpomoea aquetica[J].Journal of Liaoning University:Natural Science Edition,2004,31(3):275-278.
[19]刘广发,黄志荣.空心菜的组织培养[J].厦门大学学报:自然科学版,2001,40(s2):165-167.Liu GF,Huang ZR.The tissue culture of Ipomoea aquatica[J].Journal of Xiamen University:Natural Science,2001,40(s2):165-167.
[20]梁国婷.生长素和细胞分裂素调节拟南芥离体器官的再生[D].山东:山东农业大学,2009.
[21]龚一富,高峰,张平波.我国甘薯离体培养研究进展[J].作物研究,1998,2(19):46-48.Gong YF,Gao F,Zhang PB.Research progress on in vitro culture of sweet potato in China[J].Crop Research,1998,2(19):46-48.
[22]Schroeder HE,Schotz AH,Wardleyrichardson T,Spencer D,Higgins T.Transformation and regeneration of two cultivars of pea(Pisum sativum L.)[J].Plant Physiol,1993,101(3):751-757.
[23]Ashok Kumar HG,Ku AT,Yeh KW.An efficient and rapid plant regeneration system for sweet potato Ipomoea batatas(L.)Lam.[J].Eur J Hortic Sci,2007,72(2):85-89.
[24]孔祥生,张妙霞,陈明灿.生长素类物质对甘薯茎尖分生组织培养成苗的影响[J].山地农业生物学报,2003,22(2):114-116.Kong XS,Zhng MX,Chen MC.Effects of auxins on plantlets regeneration from subcultured tip meristem in sweet potato[J].Journal of Mountain Agriculture and Biology,2003,22(2):114-116.
[25]林珲.花椰菜高频离体再生体系的构建[J].分子植物育种,2017,15(12):5060-5064.Lin H.In vitro efficient regeneration system of cauliflower[J].Molecular Plant Breeding,2017,15(12):5060-5064.
[26]唐君,张华,刘亚菊.外源激素对甘薯不同品种茎尖培养与植株再生影响[J].江苏农业研究,2001,22(3):22-25.Tang J,Zhang H,Liu YJ.The effects on varieties and hormones on sweet potato meristem culture and plantlet regeneration[J].Jiangsu Agricultural Research,2001,22(3):22-25.
[27]张军云,张钟,张建康,陈桂芬.紫甘薯组织培养快繁技术研究[J].中国农学通报,2014,30(4):96-100.Zhang JY,Zhang Z,Zhang JK,Chen GF.Study on technology of rapid propagation in tissue culture of purple sweet potato[J].Chinese Agricultural Science Bulletin,2014,30(4):96-100.
[28]Santa-Maria M,Pecota KV,Yencho CG,Allen G,Sosinski B.Rapid shoot regeneration in industrial‘high starch’sweet potato(Ipomoea batatas L.)genotypes[J].Plant Cell Tiss Org,2009,97(1):109-117.
[29]龚一富,霍静.萘乙酸和6-苄基腺嘌呤对苷薯离体器官发生的影响[J].西南师范大学学报:自然科学版,2001,26(4):443-447.Gong YF,Hu J.Effects of naphthaleneacetic acid and 6-Benzyl adenine on the organogenesis of sweet potato[J].Journal of Southwest China Normal University:Natural Science,2001,26(4):443-447.
[30]王常芸,李晓亮,辛国胜,王建玲,王冬梅.不同品种甘薯茎尖脱毒快繁技术优化研究[J].农学学报,2015,5(7):19-23.Wang CY,Li XL,Xin GS,Wang JL,Wang DM.Research on virus-free and rapid propagation technology of sweet potato meristem culture[J].Journal of Agriculture,2015,5(7):19-23.
[31]王霞霞,马晖玲,曹冬梅.大花萱草‘香宝’再生体系的建立[J].甘肃农业大学学报,2014,49(4):136-142.Wang XX,Ma HL,Cao DM.Establishment of regeneration system of Hemeroallis middendorfii Trautv.et Mey‘Sweet Treasure’[J].Journal of Gansu Agricultural University,2014,49(4):136-142.
[32]王金祥,严小龙,潘瑞炽.不定根形成与植物激素的关系[J].植物生理学报,2005,41(2):133-142.Wang JX,Yan XL,Pan RZ.Relationship between adventitious root formation and plant hormones[J].Plant Physiology Communications,2005,41(2):133-142.
[33]席银凯,王元忠,黄衡宇,钱子刚.滇龙胆丛芽高效诱导与植株再生体系的建立[J].中草药,2018,49(6):1398-1404.Xi YK,Wang YZ,Huang HY,Qian ZG.Efficient induction system of adventitious shoot and plantlet regeneration in Gentiana rigescens[J].Chinese Traditional and Herbal Drugs,2018,49(6):1398-1404.
[34]Collado R,Veitía N,Bermúdez-Caraballoso I,García LR,Torres D,et al.Efficient in vitro plant regeneration via indirect organogenesis for different common bean cultivars[J].Sci Hortic,2013,153(3):109-116.
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