Polyphasic Taxonomy Of Three New Prokaryotic Microorganisms And The Effects Of Different Nutritional Factors On The Separation Of New Taxonomy Microbes

The thesis includes two parts: The first part is identification and polyphasic taxonomy study of strains HBUM179779 T,HBUM179663T and 3-BNT.The other part is to isolate new taxonomy microbes by adding 6 different nutritional factors in VL55 medium from soils by conventional dilution and large dilution.Obtain the following results:Strain HBUM179779 T is Gram-negative,short rods and its belongs to the family Chitinophagaceae.Single cell is measuring 0.2-0.4 ?m�1.4-1.6 ?m with peripheral flagellums.The 16 S rRNA gene sequence analysis of strain HBUM179779 T indicated most similarity to Pseudoflavitalea rhizosphaerae T16R-265T(94.46% 16 S rRNA gene sequence similarity).The DNA G+C content of strain HBUM179779 T was 47.1mol%.The predominant menaquinones was MK-7.The major cellular fatty acids were iso-C15:1 G,iso-C15:0 and iso-C17:0 3-OH.The polar lipid pattern comprised phosphatidyl ethanolamine and two unidentified lipids.Strain HBUM179663 T belongs to the genus Herbiconiux,its Cells are Gram-positive,short rods,asporogenous and non-motile.Single cell is measuring0.4-0.5 ?m�1.4-1.6 ?m.16 S rRNA gene sequence analysis indicated that strain HBUM179663 T was closely related to Herbiconiux solani K134/01T(96.65% 16 S rRNA gene sequence similarity).The DNA G+C content of strain HBUM179663 T was 75.7 mol%.The predominant menaquinones was MK-11.The major cellular fatty acids of strain HBUM179663 T were anteiso-C15:0,anteiso-C17:0 and C16:0.The polar lipid pattern comprised Phospholipids,phosphatidyl methyl ethanolamine,phosphatidyl ethanolamine and diphosphatidylglycerol.A Gram-negative,rod-shaped,non-spore-forming bacterial strain,3-BNT,was belonged to the family Burkholderiaceae.Cells are 1.8�2.1?m in length and0.4�0.6?m in diameter,and motile by means of a single polar flagellum.The 16 S rRNA gene sequence of strain 3-BNT showed most similarity to Burkholderia ubonensis CIP 107078T(92.83% 16 S rRNA gene sequence similarity).The DNA G+C content of strain 3-BNT was 42.4 mol%.The predominant isoprenoid quinone isubiquinone Q-5,Q-6 and Q-8.The principal fatty acids of strain 3-BNT were C16:0,C18:1?7c and cyclo C17:0.The predominant polar lipids profile containing diphosphatidyl glycerols,phosphatidyl ethanolamine and phosphatidylglycerol.In this study,415 pure cultures were isolated by adding six different nutritional factors in VL55 medium from three soil samples of Hebei,Qinghai and Yunnan,which isolated new taxonomy microbes by conventional dilution and large dilution.Hebei soil samples by conventional dilution isolated 72 strains were belong to 14 genus,and through large dilution isolated 52 strains were belong to 22 genus.The 37 strains of Qinghai soil samples were belong to 9 genus and 50 strains were belong to17 genus,which were isolated by conventional dilution and large dilution,respectively.The 71 strains of Yunnan soil samples were isolated by conventional dilution belonged to 23 genus,and the 133 isolates isolated by large dilution belonged to 15 genus.16 S rRNA gene sequence analysis indicated that strain HBUM200206 was closely related to Dongia mobilis LM22T(94.58% 16 S rRNA gene sequence similarity).So it may belong to the family Rhodospirillaceae.The sequence similarity of the 16 S rRNA gene of the strains HBUM200161,HBUM200202,HBUM200239,HBUM200240,HBUM200352,HBUM200424,HBUM200427,HBUM200448,HBUM200452,HBUM200488,HBUM200497 and HBUM200503 with the closest strains were less than 98.65%,which could be a potential new species.