Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii
- PDF / 752,270 Bytes
- 10 Pages / 595.276 x 841.89 pts (A4) Page_size
- 101 Downloads / 168 Views
Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii ZHANG Si1,2, LIU Cui2, JIN Yuemei2, CHI Shan2, TANG Xianming3, CHEN Fuxiao3, FANG Xu4, LIU Tao2* 1
National Marine Environmental Forecasting Center, Beijing 100081, China College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China 3 Hainan Provincial Fisheries Research Institute, Haikou 570204, China 4 School of Life Science, Shandong University, Jinan 250100, China 2
Received 16 April 2013; accepted 12 November 2013 ©The Chinese Society of Oceanography and Springer-Verlag Berlin Heidelberg 2014
Abstract 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°C 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°C, 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. Key words: Kappaphycus alvarezii, protoplast, regenerated plantlet, callus-like, young shoots Citation: Zhang Si, Liu Cui, Jin Yuemei, Chi Shan, Tang Xianming, Chen Fuxiao, Fang Xu, Liu Tao. 2014. Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii. Acta Oceanologica Sinica, 33(10): 114–123, doi: 10.1007/s13131-014-0546-y
1 Introduction Kappaphycus alvarezii belongs to Rhodopyta, Florideae, Gigartinales, Solieriaceae and Kappaphycus. It is widely distributed in tropical and subtropical sea areas. Kappaphycus seaweed is the main raw material to produce carrageenan, thus it is widely used in textiles and food. In addition, owing to its high pharmaceutical value (Jiang et al., 2008), economic value as well as ecological significance (Tseng, 1983; Zeng, 2001), there is a great demand for Kappaphycus both in the domestic and foreign markets. As a crucial biological material for research purpose, protoplast without cell wall obstacles can be widely used in the study of cell biology, somatic cell crossbreeding, genetic transformation and gene positioning (Wang and Yao, 2009). Meanwhile, the culture of protoplasts is an important plant somatic cell breeding technology. By now, the photoplasts isolation and cultivation have been performed on more than 30 algae including Rhodophyta, Chlorophyta and Phaeophyta (Wang et al., 1996; Wu, 1988; Chu et al., 1998; Wang et al., 2007; Zhang, 1983; Hu, 2006; Araki and Morishita, 1990; Ma et al., 1998; Chen and Chiang, 1994; Chen, 1993; Reddy and Fujita, 1991; Della et al., 1991; Reddy et al., 1992; Xie and Ma, 2004; Kim et al., 2002; Zha and Kloareg, 1996; Zhang, 1991). So
Data Loading...
