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

对于该研究，使用以下物种;作为琼脂植物的江蓠、Ahnfeltiopsis paradoxa、Gelidium subfastigiatum和作为角叉菜植物的角叉菜、角叉菜yendoi。用琼脂酶、纤维素酶和果胶酶混合酶（溶于0.7M甘露醇MES高压灭菌海水中）在25 ℃、60- 120 min条件下制备了虫孔江蓠和奇异拟杆藻的原生质体，获得了可用于融合的原生质体。但由于卡拉胶酶难以获得，卡拉胶原生质体的制备一直没有成功。从冰冻三尺症状中分离出的真菌提取物有可能应用于长柄卡帕藻，但不完全。在PES培养基中加入植物生长调节剂（生长素组：2，4-D、IAA、IBA、IPA、pCA、α-NAA、β-NAA;细胞分裂素组：BAP、KIN、ZEA;赤霉素组：GA_3）进行组织培养。在2，4-D处理下，异花龙须菜仅在8和15口处的髓质中观察到新芽的形成，而江蓠在8和15口处的外周层和髓质中观察到新芽的形成。在石花菜中，所有植物生长调节剂均在8和15□处的外周层和髓质中观察到这种现象，特别是2，4-D和IAA。角叉菜属（Chondrus spp.结果表明，用植物生长调节剂进行组织培养，对海藻的种苗生产和品种生产是有效的，但对附着器官的形成却不完全。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.