Zhang Si, Liu Cui, Jin Yuemei, Chi Shan, Tang Xianming, Chen Fuxiao, Fang Xu, Liu Tao. Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii[J]. Acta Oceanologica Sinica, 2014, 33(10): 114-123. doi: 10.1007/s13131-014-0546-y
Citation: Zhang Si, Liu Cui, Jin Yuemei, Chi Shan, Tang Xianming, Chen Fuxiao, Fang Xu, Liu Tao. Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii[J]. Acta Oceanologica Sinica, 2014, 33(10): 114-123. doi: 10.1007/s13131-014-0546-y

Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii

doi: 10.1007/s13131-014-0546-y
  • Received Date: 2013-04-16
  • Rev Recd Date: 2013-11-12
  • In this study, protoplasts were successfully isolated from Kappaphycus alvarezii using snail enzymes, abalone enzymes and cellulase. The optimum enzymic ratio was fixed to be 20% of abalone enzyme, 12% of cellulase and the osmotic stabilizer was 2.0 mol/L glucose. The optimum enzymic hydrolysis conditions were found to be dark enzymolysis at 30℃ continuing for 4.0 h. The resultant density and yield of protoplasts achieved 32.60×104 mL-1, 65.20×104 g-1 tissue for Kappaphycus alvarezii. Finally, under the temperature of 20℃, light intensity of 1 500-2 000 lx and photoperiod of 12 h/d, two developmental pathways were investigated: (1) callus-like cell mass and regenerated plantlet occurred on protoplast; (2) young shoots and calluslike cell mass occurred in tissue blocks after enzymolysis.
  • loading
  • Araki T, Morishita T. 1990. Fusion of protoplasts from wild type Porphyra yezoensis and green type P. tenera thalli (Phodophyta.). Nippon Suisun Gakkaishi, 56: 1161-1161
    Bold H C, Wynne M J. 1978. Introduction to the Algae: Structure and Reproduction. Englewood Cliffs, N J: Prentice-Hall, 577-578
    Chen Juqi. 1993. Studies on caloglossa protoplast isolation by using the viable enzyme. Journal of Xiamen Fisheries College (in Chinese), 15(1): 1-7
    Chen L C M. 1987. Protoplast morphogenesis of Porphyra leucosticta in culture. Botanica Marina, 30(5): 399-404
    Chen Y C, Chiang Y M. 1994. Development of protoplasts from Grateloupia sparsa and G. flicina (Halymeniaceae, Rhodophyta). Botanica Marina, 37(4): 361-366
    Chu Jiansong, Liu Wanshun, Zhang Zhaoyang. 1998. A preliminary study on the isolation and cultivation of protoplasts from G.Verrucosa. Marine Science Bulletin, 17(6): 17-26
    Dai Jixun, Bao Zhenmin, Tang Yanlin, et al. 1988. Studies on isolation of the thallodic cells of porphyra with enzymes and cultivation of these cells. Chinese Journal of Biotechnology, 4(2): 133-137
    Della P F, Balestri E, Cinelli F. 1991. Preliminary results on production of protoplasts from two green marine algae: Enteromorpha linza (L.) J. Agardh and Ulva rigida C. Agradb. International Journal of Biological Macromolecules, 17: 553-560
    Guo Zian, Ji Hongwu, Shao Haiyan, et al. 2008. Optimization of technology for enzymatic degradation of sea weed Eucheuma striatum. Fisheries Science (in Chinese), 27(7): 348-351
    Hu Yinmao. 2006. The morphogeny of callus from single somatic cell clone cultivation of Porphyra haitanensis. Journal of Taizhou University (in Chinese), 28(3): 67-69
    Hu Yuanjie, Yin Changsong, Sun Yuping. 2000. Preparation and analysis of Alginate Lyases from Abalon. Fisheries Science & Technology Information (in Chinese), 27(6): 256-258
    Hurtado A Q, Biter A B. 2007. Plantlet regeneration of Kappaphycus alvarezii var. adik-adik by tissue culture. Journal of Applied Phycology, 19(6): 783-786
    Hurtado A Q, Yunque D A, Tibubos K, et al. 2009. Use of Acadian marine plant extract powder from Ascophyllum nodosum in tissue culture of Kappaphycus varieties. Journal of Applied Phycology, 21(6): 633-639
    Inoue A, Mashino C, Kodama T, et al. 2011. Protoplast preparation from Laminaria japonica with recombinant alginate lyase and cellulase. Marine Biotechnol, 13(2): 256-263
    Jiang Zhenyou, Liu Yue, Tang Yu, et al. 2008. Effect of Seleno-polysaccharide of Eucheuma on growth and apoptosis of hepatoma carcinoma cell strain. Journal of Sun Yat-Sen University: Medical Sciences (in Chinese), 29(3): 270-277
    Kim G H, Klotchkova T A, West J A. 2002. From protoplasm to swarmer: regeneration of protoplasts from disintegrated cells of the multicellular marine green alga Microdictyon umbilicatum (Chlorophyta). Journal of Philosophy, 38(1): 174-183
    Leila H, Nair S Y, Daniela M K, et al. 2008. Callus induction and micropropagation improved by colchicine and phytoregulators in Kappaphycus alvarezii (Rhodophyta, Solieriaceae). Journal of Applied Phycology, 20(5): 653-659
    Lillo C. 1989. Effects of media components and environmental factors on shoot formation from protoplast derived calli of solanum iuberosum. Plane Cell, Tissue and Organ Culture, 19(2): 103-111
    Liu Chengsheng, Liu Chenguang, Meng Xianghong, et al. 2002. Preparation and characteristics of snail enzymes. Journal of Wuhan University (Natural Science Edition) (in Chinese), 48(2): 243-248
    Liu Shuxia, Zou Dinghui, Xu Juntian. 2007. Tissue culture and its application in marine macroalgae. Plant Physiology Communications (in Chinese), 6(3): 605-611
    Ma Lingbo, Wang Sujuan, Huang Yipeng, et al. 1998. Isolation using bacterial enzyme and indoor artificial breeding of somatic cells from porphyra yezoensis. Journal of Fisheries of China (in Chinese), 22(S1): 24-30
    Mussio I, Rusig A M. 2009. Morphogenetic responses from protoplasts and tissue culture of Laminaria digitata (Linnaeus) J. V. Lamouroux (Laminariales, Phaeophyta): callus and thalloid-like structures regeneration. Journal of Applied Phycology, 21(2): 255-264
    Reddy C R K, Fujita Y. 1991. Regeneration of plantlets from Enteromorpha (Ulvales, Chlorophyta) protoplasts in axenic culture. Journal of Applied Phycology, 3: 265-275
    Reddy C R K, Lima M, Fujita Y. 1992. Induction of fast growing and morphologically different strains through intergenetic protoplast fusions of Ulva and Enteromorpha (Ulvales, Chlorophyta). Journal of Applied Phycology, 4(1): 57-65
    Saga N, Sakai Y. 1984. Isolation of protoplasts from Laminaria and Prophyra. Bulletin of the Japanese Society for the Science of Fish, 50(6): 1085-1085
    Salvador R C, Serrano A E. 2005. Isolation of protoplasts from tissue fragments of Philippine cultivars of Kappaphycus alvarezii (Solieriaceae, Rhodophyta). Journal of Applied Phycology, 17(1): 15-22
    Tang Yanlin. 1982. Isolation and cultivation of the vegetative cells and protoplasts of Porphyra suborbiculata Kjellm. Journal of Shandong College of Oceanology (in Chinese), 12(4): 37-50
    Tseng C K. 1983. Common Seaweeds of China. Beijing: Science Press Wang Sujuan. 1994. Seaweed Biotechnology (in Chinese). Shanghai: Shanghai Science and Technology Press Wang Sujuan, Sun Yunlong, Ma Lingbo, et al. 1996. The isolation and culture of protoplasts from Sargassum fusiforme (Phaeophyta, Sargassaceae). Journal of Fisheries of China (in Chinese), 20(3): 241-249
    Wang Mingzi, Wang Shifeng, Shi Qiaoqin, et al. 2007. Regeneration of protoplasts from Porphyridium cruentum and its variation for breeding. Journal of Fujian Normal University (Natural Science Edition) (in Chinese), 23(1): 65-69
    Wang Fude, Xia Zhenao. 1981. Fluorescent observation of plant cell walls. Plant Physiology Communications (in Chinese), 1: 59-61
    Wang Sujuan, Xu Zhidong, Chen Changsheng. 1985. Tissue culture of medulla and regeneration of the fragments of Solieria tenuis. Marine Fisheries (in Chinese), 7(4): 155-159
    Wang Li, Yao Zhanjun. 2009. Advances on the preparation and vitality testing for plant protoplast. Biotechnology Bulletin (in Chinese), (S1): 46-50
    Wu Shaobo. 1988. Isolation and culture of protoplasts from Undaria pinnatifida (Harv) Suringar. Journal of Ocean University of Qingdao (in Chinese), 18(2): 57-65
    Xie Enyi, Ma Jiahai. 2004. Isolation and cultivation of protoplasts of Monostroma nitidum (Chlorophyta, Monostromaceae). Journal of Fisheries of China (in Chinese), 28(1): 62-67
    Xu Xinghong, Wang Ping, Zhao Suzhen. 2007. Study on the tool enzymes of isolating protoplasts of Porphyra yezoensis. Journal of Huaihai Institute of Technology (Natural Sciences Edition (in Chinese), 16(3): 66-69
    Yu Changying, Shen Songdong, Dai Jixun, et al. 2000. Extraction of seaweed cell wall decomposing enzyme from Chiton sp. and Patella sp. Marine Science Bulletin (in Chinese), 19(5): 92-96
    Yu Xiaoling, Li Chunqiang, Peng Ming. 2009. Advances on the rsearch of potoplast technology. Chinese Agricultural Science Bulletin (in Chinese), 25(8): 22-26
    Zeng Guangxing. 2001. Artificial breeding technology of Eucheuma striatum. Journal of Aquaculture (in Chinese), (3): 5-7
    Zha Xiangdong, Kloareg B. 1996. Protoplast isolation from sporophytes of three brown algae. Journal of Anhui Agricultural Sciences (in Chinese), 24(2): 184-185, 187
    Zhang Dali. 1983. Studies on the protoplast preparation, culture and fusion of somatic cells from two species of green algae-Ulva linza and Monostroma angicava Kjellm. Journal of Shandong College of Oceanlogy (in Chinese), 13(1): 57-66
    Zhang Quanqi. 1991. Studies on theisolation, culture and regeneration of protoplasts from Chondrus ocellatus Holm. Journal of Ocean University of Qingdao (in Chinese), 21(1): 52-62
    Zhao Dake, Wang Jingqiao, Chen Wei, et al. 2009. Effect of the gaining of protoplast, culture conditions and masures on the plant protoplast to plantlet system construction. Journal of Yunnan University (in Chinese), 30(S2): 403-408
    Zhao Sufen, Zhuang Chengji. 1999. Regeneration of the fragments of two Gracilaria. Journal of Zhanjiang Ocean University (in Chinese), 19(2): 25-29
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Article Metrics

    Article views (1576) PDF downloads(1649) Cited by()
    Proportional views
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return