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；龙须菜 (Gracilaria vermiculopylla)、Ahnfeltiopsis paradoxa、石花菜 (Gelidium subfastigiatum) 为琼脂植物，角叉菜为角叉菜 (Chondrus ocellatus)、角叉菜 (Chondrus Yandoi)。采用混合酶(琼脂酶、纤维素酶、果胶酶溶解于0.7M甘露醇MES高压灭菌海水中)在25℃、60-120min下制备江蓠和Ahnfeltiopsis paradoxa原生质体，获得可用于融合的原生质体。然而，由于卡拉胶酶难以获得，来自卡拉胶植物的原生质体并没有成功。可以使用从卡帕藻（Kappaphycus alvarezii）冰冰症状中分离出来的真菌提取物，但不完全。需要更多的研究。使用植物生长调节剂（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）溶解在PES培养基中进行组织培养。 Ahnfeltiopsis paradoxa仅在8和15℃的髓质中观察到新芽形成，而Gracilaria vermiculopylla在2、4-D中在8和15℃的外周层和髓质中观察到新芽形成。在所有植物生长调节剂中，特别是在2、4-D和IAA中，在8和15℃下，在Gelidium subfastigiatum的外周层和髓质中也观察到了这种情况。在角叉菜属中。髓质中形成新芽，2, 4-D效果良好。从这些结果来看，使用植物生长调节剂的组织培养对于海藻的幼苗和品种生产非常有效，但粘附器官的形成不完全。因此有必要继续对此事进行研究。

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.