268 海南师范学院学报(自然科学版) 2005年 [25 GROSSER J W. CHANDLER J L. Aseptic isolation of protoplasts from Citrus, Poncirus, Citrus XRbncirus hybrids and severinia for use in somatic hybridization experiments J]. Scientia hortic, 1989 31: 253-257 [26]卫志明,许智宏,许农,等.悬铃木叶肉原生质体培养再生植株[]植物学报,1991,311):813-818 [27]马锋旺,李嘉瑞抗氧化剂对杏和中国李原生质体培养的影响[]西北农业学报,1998,26(6):10·13 [28]赵颖.梁海曼.植物原生质体培养及有关生理基础问题卩].广西植物,1994,14(1):74-80 [29]肖尊安沈德绪,林伯年美味猕猴桃子叶愈伤组织的原生质体培养和再生植株卩].武汉植物学研究,1993,113):247 [30] OCHATT SJ, CHEVREAU E, GALLETM. Orgamgenesis fom 'Passe Crassane and ' Old Home pear( Pyrus Commnis L )proto- plasts and isoenzymatic trueness- to-type of the regenerated plants [ ]. The Apple Genet, 1992, 83: 1 013.1 018 [31]吴家道杨剑波,向太和,等水稻原生质体高效培养技术的研究[]安傲农业科学,199422(4):305-309 32 HUANCARUNA- PERALES E, SCHIEDER O. ant regene at on irom protoplasts of apple [J]. Pant cell, Tissue &Organ Cultur 1993,34:71-76 [33 CHEN M H, CHEN CC. Pant regeneration from Carica protoplasts[J]. Pant Cell Reports, 1992, 11: 404-407. [34 LI R B. Protoplast culture and plant regeneration of indica rice by nurse culture and capsulation technique J]. Guang xi Science 1998,5(3):230-236 [35 ABDULLAH R, COCKING E C, THOMPSON J A Hilicient plant regeneration from rice protoplats through somatic embryogensisJI Bio thchnology,19864:1087-1090 [36]颜昌敬.植物组织培养手册[M].上海:科学技术出版社,1990 [37 VASLL J K Developing cell and tissue culture systems for improvement of cereal and grass crops[J]. J Hant Physoil, 1987, 128: 192 [38]贾敬芬.禾谷类植物原生质体培养和融合[A].孙敬三,陈维伦.植物生物技术和作物改良[C].北京:中国科学技术出版 社,1990.71-82 [39]黄培铭,颜昌敬。大麦叶肉原生质体的分离与培养初报[].上海农业学报,190,6(4):17 [40]孙勇如,李文彬,黄美娟,等.枸杞的原生质体生成愈伤组织卩]植物学报,1982,24(5):477-479 [41 KAO KN. A method for higher- frequency intergenic fusion of plant protoplast [J]. Panta, 1974, 115: 355- 367 [42 SENDA M. Pant cell physiol [] Pant cell Rep, 1979, 20: 1 441- 1 443 [43]汪和睦,谢廷栋.细胞电穿孔、电融合、电刺激—原理技术及应用[M]天津:天津科学技术出版社,2000 [44]诸葛强,黄敏仁,王明庥.杨树体细胞融合研究[].南京林业大学学报200,24(2):6·10 [45]黄白渠.植物体细胞遗传学简明教程[M],长春:东北师范大学出版社,199 [46 SUNDBER E, GIMEIUS KA. Method for production of interspecific hybrids within B. rassiceae via somatic hybridization using resyn theis is of B. rassica napusas a model[J]. Pant Sci, 1986, 43: 155-162 [47]蔡起贵美味猕猴桃原生质体再生植株无性系变异的研究卩].植物学报,199,34:822-888 48]邓秀新,刘继红.果树遗传转化及基因组转移研究进展[]武汉植物学研究,1996,14(4):357·369 49 PUITE KJ. Progress in plant protoplast research[J. Physiol plant 1992, 85: 403-410 Advances in researches on protoplast of plants I Chun-yan, LI Nrya, CHEN Shao-liang College d Biological Sciences and Technology, Beijing Forestry University, Bejing 100083,Chi Abstract The isolation. cu nd fusion technology of plant protoplast have been developing rapidly over the past twenty years In this peper, we concisely summa he protoplast isc culture fusion and plant regeneration. Besides, existing problems and fur- ther development were also discussed Key words: protoplast; isolation; culture; fusion; plant regeneration[25 ] GROSSER J W. CHANDLER J L. Aseptic isolation of protoplasts from Citrus , Poncirus , Citrus×Poncirus hybrids and severinia for use in somatic hybridization experiments [J ]. Scientia hortic ,1989 ,31 : 253 - 257. [26 ] 卫志明 ,许智宏 ,许 农 ,等. 悬铃木叶肉原生质体培养再生植株[J ]. 植物学报 ,1991 ,33(11) : 813 - 818. [27 ] 马锋旺 ,李嘉瑞. 抗氧化剂对杏和中国李原生质体培养的影响[J ]. 西北农业学报 ,1998 ,26(6) :10 - 13. [28 ] 赵 颖 ,梁海曼. 植物原生质体培养及有关生理基础问题[J ]. 广西植物 ,1994 ,14 (1) : 74 - 80. [29 ] 肖尊安 ,沈德绪 ,林伯年. 美味猕猴桃子叶愈伤组织的原生质体培养和再生植株[J ]. 武汉植物学研究 ,1993 ,11(3) : 247 - 251. [ 30 ] OCHATT SJ , CHEVREAU E , GALLET M. Organogenesis from‘Passe Crassane’and‘Old Home’pear (Pyrus Commnis L. ) proto2 plasts and isoenzymatic trueness - to - type of the regenerated plants [J ]. The Apple Genet ,1992 ,83 :1 013 - 1 018. [31 ] 吴家道 ,杨剑波 ,向太和 ,等. 水稻原生质体高效培养技术的研究[J ]. 安徽农业科学 ,1994 ,22(4) :305 - 309. [32 ] HUANCARUNA - PERALES E , SCHIEDER O. Plant regeneration from protoplasts of apple [J ]. Plant cell、Tissue&Organ Culture , 1993 ,34 :71 - 76. [33 ] CHEN M H , CHEN C C. Plant regeneration from Carica protoplasts[J ]. Plant Cell Reports ,1992 ,11 :404 - 407. [34 ] LI R B. Protoplast culture and plant regeneration of indica rice by nurse culture and capsulation technique [J ]. Guang xi Science , 1998 ,5 (3) :230 - 236. [35 ] ABDULLAH R , COCKING E C , THOMPSON J A. Efficient plant regeneration from rice protoplats through somatic embryogensis[J ]. Bio Thchnology ,1986 ,4 :1 087 - 1 090. [36 ] 颜昌敬. 植物组织培养手册[M]. 上海 :科学技术出版社 ,1990. [37 ] VASILL J K. Developing cell and tissue culture systems for improvement of cereal and grass crops[J ]. J Plant Physoil ,1987 ,128 :193 - 218. [38 ] 贾敬芬. 禾谷类植物原生质体培养和融合[A]. 孙敬三 , 陈维伦. 植物生物技术和作物改良[ C]. 北京 :中国科学技术出版 社 ,1990. 71 - 82. [39 ] 黄培铭 ,颜昌敬. 大麦叶肉原生质体的分离与培养初报[J ]. 上海农业学报 ,1990 ,6(4) : 17 - 20. [40 ] 孙勇如 ,李文彬 ,黄美娟 ,等. 枸杞的原生质体生成愈伤组织[J ]. 植物学报 ,1982 ,24(5) : 477 - 479. [41 ] KAO K N. A method for higher - frequency intergenric fusion of plant protoplast [J ]. Planta ,1974 ,115 :355 - 367. [42 ] SENDA M. Plant cell physiol [J ]. Plant cell Rep ,1979 ,20 :1 441 - 1 443. [43 ] 汪和睦 ,谢廷栋. 细胞电穿孔、电融合、电刺激 ———原理技术及应用[M]. 天津 : 天津科学技术出版社 ,2000. [44 ] 诸葛强 ,黄敏仁 ,王明庥. 杨树体细胞融合研究[J ]. 南京林业大学学报 ,2000 , 24 (2) : 6 - 10. [45 ] 黄白渠. 植物体细胞遗传学简明教程[M]. 长春 : 东北师范大学出版社 ,1991. [46 ] SUNDBER E , GIMELIUS KA. Method for production of interspecific hybrids within B. rassiceae via somatic hybridization using resyn2 theis is of B. rassica napusas a model[J ]. Plant Sci ,1986 ,43 :155 - 162. [47 ] 蔡起贵. 美味猕猴桃原生质体再生植株无性系变异的研究[J ]. 植物学报 ,1992 ,34 :822 - 888. [48 ] 邓秀新 ,刘继红. 果树遗传转化及基因组转移研究进展[J ]. 武汉植物学研究 ,1996 ,14(4) :357 - 369. [49 ] PUITE KJ . Progress in plant protoplast research[J ]. Physiol plant ,1992 ,85 :403 - 410. Advances in researches on protoplast of plants LI Chun2yan , LI Ni2ya , CHEN Shao2liang ( College of Biological Sciences and Technology , Beijing Forestry University , Beijing 100083 , China) Abstract : The isolation , culture and fusion technology of plant protoplast have been developing rapidly over the past twenty years. In this peper , we concisely summarized the protoplast isolation , culture , fusion and plant regeneration. Besides ,existing problems and fur2 ther development were also discussed. Key words: protoplast ; isolation ; culture ; fusion ; plant regeneration 862 海南师范学院学报(自然科学版) 2005 年