Elucidation of symbiotic phenomenon and specific diversity of symbionts in Anthocerotae (Bryophyte).

本研究旨在阐明苔藓藓属植物的共生现象和特定多样性。以日本不同生境的野生石斛为研究对象，对其与寄主相关的共生体物种多样性进行了研究。结果表明，同一个体内共生蓝藻的物种多样性在卡罗氏单胞菌上得到了充分证实，而在鞭毛单胞菌上却很少观察到这种现象。与宿主相关的共生蓝藻被鉴定为Nostoc sphaericum， N.sp。1, N.sp。2, N.sp.3。这种现象的根源在于当下的学习。采用分离法分离寄主和共生体，分别进行培养。在Knop培养基上用成熟孢子培养分离寄主，用无菌针从寄主菌体中分离共生体，在MDM琼脂培养基上培养。两株菌株均在培养室、20℃、12h/12明暗循环培养条件下生长良好。为了重建共生状态，我们将从孢子中培养出来的卡罗莉花的成熟菌体放在添加了BBM溶液培养基的滤纸上。之后，尽快混合共生菌（Nostoc sp.）的激素态细丝。大约一周后开始重建，大约一个月后形成大容量。本研究对共生关联进行了完善的重构。作为重建，在重建的每个发育阶段对宿主进行组织切片。这些切片的结果清楚地表明，激素通过腹侧小孔进入宿主组织。之后，激素在腔隙中生长并形成腔体。

This investigations were made to elucidate symbiotic phenomenon and specific diversity in Anthocerotae, Bryophyte. Wild Anthocerotae, especially Phaeoceros carolinianus and Megaceros flagellaris collected from various habitats in Japan were studied on species diversity of symbionts associated with host. As the results, it was cleared that species diversity of symbiotic cyanobacteria in one individual was abundantly confirmed on P.carolianus, but on M.flagellaris, this phenomenon was rarely observed. Symbiotic cyanobacterias associating with hosts were identified as Nostoc sphaericum, N.sp.1, N.sp.2, N.sp.3. The root of this phenomenon is studying in the present time. On P.carolinianus, host and symbionts were separated by isolation methods, and cultured respectively. The separate of host was made by mature spore culture in Knop medium, and symbionts were isolated from thalli of host by axenic needle and cultured on MDM agar medium. Both strains grown under good state in culture room, 20℃, 12h/12 light and dark cycles culture condition. As reconstruction of symbiotic state, mature thalli of P.carolianua grown from spore were put on filter papers added BBM solution medium. After that, hormogon state filaments of symbiont (Nostoc sp.) were mixed as possible as soon. Their reconstruction was started after about one weeks and large cabities were formed after about one monthe. In this study, perfect reconstruction of symbiotic association was made. As reconstruction, Tissue sections of the host were made as each developmental stage of reconstruction. As results of these sections, it was clear that hormogons inversion into host tissue after passing through small pore in ventral portion. After that, hormogons were vigorously grown at the lacuna and made cavities.