Magnani G S, Didonet C M, Cruz L M, Picheth C F, Pedrosa F O, Souza E M
Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Paraná, Curitiba, PR, Brasil.
Genet Mol Res. 2010 Feb 9;9(1):250-8. doi: 10.4238/vol9-1gmr703.
Endophytic bacteria live inside plant tissues without causing disease. Studies of endophytes in sugarcane have focused on the isolation of diazotrophic bacteria. We examined the diversity of endophytic bacteria in the internal tissues of sugarcane stems and leaves, using molecular and biochemical methods. Potato-agar medium was used to cultivate the endophytes; 32 isolates were selected for analysis. DNA was extracted and the 16S rRNA gene was partially sequenced and used for molecular identification. Gram staining, catalase and oxidase tests, and the API-20E system were used to characterize the isolates. The strains were divided into five groups, based on the 16S rRNA sequences. Group I comprised 14 representatives of the Enterobacteriaceae; group II was composed of Bacilli; group III contained one representative, Curtobacterium sp; group IV contained representatives of the Pseudomonadaceae family, and group V had one isolate with an uncultured bacterium. Four isolates were able to reduce acetylene to ethylene. Most of the bacteria isolated from the sugarcane stem and leaf tissues belonged to Enterobacteriaceae and Pseudomonaceae, respectively, demonstrating niche specificity. Overall, we found the endophytic bacteria in sugarcane to be more diverse than previously reported.
内生细菌生活在植物组织内部而不引发疾病。对甘蔗内生菌的研究主要集中在固氮细菌的分离上。我们运用分子和生化方法,研究了甘蔗茎和叶内部组织中内生细菌的多样性。采用马铃薯琼脂培养基培养内生菌;选取32个分离株进行分析。提取DNA并对16S rRNA基因进行部分测序,用于分子鉴定。采用革兰氏染色、过氧化氢酶和氧化酶试验以及API-20E系统对分离株进行特征描述。根据16S rRNA序列,这些菌株被分为五组。第一组包括肠杆菌科的14个代表菌株;第二组由芽孢杆菌组成;第三组包含一个代表菌株,即短小杆菌属细菌;第四组包含假单胞菌科的代表菌株,第五组有一个未培养细菌的分离株。有四个分离株能够将乙炔还原为乙烯。从甘蔗茎和叶组织中分离出的细菌大多分别属于肠杆菌科和假单胞菌科,显示出生态位特异性。总体而言,我们发现甘蔗中的内生细菌比此前报道的更加多样。