关键词: 海带配子体, 消毒方式, 封口方式, 生长, 染菌, Saccharina japonica gametophyte, sterilization, flask sealing, growth, bacteria contamination

Abstract: China has cultured the largest areas of kelp all over the world. It has firstly applied the gamete clone breeding techniques to the kelp hybridization practices in history. During the germplasm conservation and amplification cultivation of Saccharina gametes, bacterial contamination usually causes great losses to Saccharina germplasm resources and could further lead to economic losses to the kelp hybridization and kelp industry. In this study, four factors were set to test their effects on the growth and bacterial contamination of Saccharina japonica gametophytes. (1) Three means for flask sealing: Parafilm significantly inhibited the growth of the gametes (p<0.05) while filter paper and filter paper membrane did not. (2) Five means for seawater sterilization: Gamete clusters in boiled seawater and suction filtered boiled seawater groups reached the largest diameters which had significant preponderance over groups of double-antibody suction filtered boiled seawater, suction filtered unboiled seawater and double-antibody suction filtered unboiled seawater (p<0.05). (3) Two environments for illumination incubator: conventional laboratories and sterile room had no significant effect on the growth of the gametes. (4) Three environments for replacing culture medium: clean bench in both conventional laboratories and sterile room had no significant difference on the growth of gametes but had significant preponderance over experiment bench in conventional laboratories (p<0.05), and replacing the culture medium on experimental bench in conventional laboratories led to a bacterial contamination rate over 60%.