Selection and cultivar production in agarophytes and carragenophytes using tissue culture and cell fusion methods

本研究使用了以下物种：细叶青藤、拟拟拟南芥、细叶青藤为食糖植物，而细叶青藤、叶青藤为食糖植物。采用琼脂酶、纤维素酶和果胶酶的混合酶（分别溶解于0.7 M甘露醇MES高压灭菌海水中），在25℃和60-120min条件下制备蠕皮草和异拟拟豆的原生质体，获得可用于融合的原生质体。然而，由于难以获得角叉菜酶，从角叉菜内生植物中提取原生质体并没有成功。从冻害症状分离的真菌中提取的物质有可能用于木耳，但不完全。有必要进行更多的研究。将植物生长调节剂（生长素组为2,4 - d、IAA、IBA、IPA、pCA、α-NAA和β-NAA，细胞分裂素组为BAP、KIN和ZEA，赤霉素组为GA3）溶解于PES培养基中进行组织培养。在2,4 - d的8、15□时，拟南毛参只在髓质中出现了新芽，而在8、15□时，虎茅的外周层和髓质中也出现了新芽。所有植物生长调节剂，特别是2,4 - d和IAA，在8和15□时均在花萼下的外周层和髓质中观察到。在球茎属植物中，新芽在髓质中形成，2,4 - d效果较好。由此可见，植物生长调节剂对海藻的组培育苗和品种生产是非常有效的，但粘附器官的形成并不完全。因此，有必要继续对这一问题进行研究。

For the study following species were used ; Gracilaria vermiculopylla, Ahnfeltiopsis paradoxa, Gelidium subfastigiatum as agarophytes and Chondrus ocellatus, Chondrus yendoi as carragenophytes. Protoplast preparations of Gracilaria vermiculopylla and Ahnfeltiopsis paradoxa were done using enzyme mixture (agarase, cellulase maacerozyme, and pectinase which were dissolved in 0.7 M mannitol MES autoclaved seawater) under 25 □and 60-120min, and usable protoplasts for fusion were obtained. However the protoplast from carrageenophytes didn't success because of the difficulty to get carragenase. It was possible to use the extracted substance fron fungi which isolated from the symptom of ice-ice on Kappaphycus alvarezii, but not complete. It is necessary more studies.Tissue culture was done using plant growth regulators( 2, 4-D, IAA, IBA, IPA, pCA, α-NAA, and β-NAA in auxine group ; BAP, KIN, and ZEA in cytokinin group and GA3 in gibberellin group) dissolved in PES medium. New bud formation in Ahnfeltiopsis paradoxa was only observed in medulla at 8 and 15□, and that in Gracilaria vermiculopylla was observed in peripheral layer and medulla at 8 and 15□in 2, 4-D. Also that in Gelidium subfastigiatum was observed in peripheral layer and medulla at 8 and 15□in all of plant growth regulators, specially in 2, 4-D and IAA. In Chondrus spp. New bud was formed in medulla and 2, 4-D gave a good results.From these results tissue culture using plant growth regulators is very effective fro the seedling and cultivar production in seaweeds, but the formation for adhesive organ was not complete. So that its is necessary to continue the study on this matter.