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

在研究中使用了以下几种植物：龙须菜、异叶龙须菜、亚高山石花菜为无菌植物，角叉菜、叶海棠为角叉菜。用混合酶(琼脂酶、纤维素酶、果胶酶溶解于0.7M甘露醇高压灭菌海水中)分别在25分钟和60-120分钟内制备江豚和反常龙须菜的原生质体，获得了可用于融合的原生质体。然而，由于卡拉胶原生质体难以获得卡拉胶酶，原生质体未能获得成功。从冰冻症状中分离出的真菌中提取的物质可以用于高山海棠，但不完全可以使用。用植物生长调节剂(生长素组为2，4-D、IAA、IBA、iPA、Pca、α-NAA、β-NAA；细胞分裂素组为BAP、KIN、ZEA；赤霉素组为赤霉素组)进行组织培养。在2，4-D中，反常龙须草的新芽形成仅在延髓中观察到，在8和15-D中，龙须菜的新芽形成在外周层和延髓中都能观察到。此外，在植物生长调节剂中，以2，4-D和IAA为代表的植物生长调节剂在第8天和第15天的外周层和延髓中也有表达。角叉菜属(Chondrus spp.)结果表明，用植物生长调节剂进行组织培养对海藻的育苗和品种生产是非常有效的，但贴壁器官的形成还不完全。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.