Morphological And Phylogenetic Study Of Three Algae Associated With The Bryophytes

We isolated three algae symbiosis with bryophytes, and performed analysis of these using both morphological characterization and molecular methods. The results were as follows:(1) Mnium cuspidatum Hedw. from the Bashuigou landscape of the Guandi Mountains, Shanxi Province, China, found to be associated with a cyanobacterium that was successfully isolated and cultivated under axenic conditions and designated as Microcoleus sp. GD based on its morphology. Phylogenetic analysis of 16S rRNA alone and of the 16S rRNA combined with the 16S-23S internal transcribed spacer (ITS) region, and the presence of an 11-bp insert in the 16S rRNA gene,indicated that Microcoleus sp. GD was as Microcoleus vaginatus. Phylogenetic analysis of the 16S rRNA gene sequences revealed that Microcoleus sp. GD formed a well-supported clade with aquatic epipelic strains of M. vaginatus from the Czech Republic and with desert strains from the USA. Microcoleus sp. GD was more closely related to the aquatic strains living at the seciment/water interface. The secondary structure of the 16S-23S ITS showed that the Box-B helix of Microcoleus sp. GD was structurally was similar to these strains but quite different from the desert strains. Furthermore, the Dl-D1’ helices of all strains varied but Microcoleus sp. GD was again most similar to the aquatic strains.(2) The algal photobiont of the moss Haplocladium strictulum (Card.) Reim. from the Guancen Mountains, Shanxi Province, China, was successfully isolated and cultivated under axenic conditions and designated as GC003. The algal photobiont of Timmiella anomala Limpr. from the Guandi Mountains, Shanxi Province, was isolated and cultivated in a similar fashion and designated as GD001. Based on morphological observations, the two isolated chrysophytes were found to be similar to Ochromonas and Poterioochromonas, but with no loricas or scales present outside the cell. Phylogenetic analysis indicated that these two algal strains are most closely related to P. malhamensis. The ability of P. malhamensis to form a lorica may thus be related to its living environment.(3) The algal photobiont of the moss Entodon obtusatus from the Guandi Mountains, Shanxi Province, China, was successfully isolated and cultivated under axenic conditions and designated as Chlorococcum sp. GD. Based on morphological observations, the isolated algae were found to be similar to Chlorococcum。Based on phylogenetic analysis of 18S rRNA, rbcL gene and ITS gene, Chlorococcum sp. GD was identified as C. sphacosum. But the original description of C. sphacosum is uncompleted. And in the present classification, C. sphacosum is synonymous with C. minutum. This study found that C. minutum is diverse on phylogeny tree. The habitat of C. sphacosum is related to bryophyte. So we suggested that C. sphacosum should be independent. At the same time, we added the character C. sphacosum.