Molecular characterisation of the diazotrophic bacterial community in uninoculated and inoculated field-grown sugarcane (Saccharum sp.)
Molecular characterisation of the diazotrophic bacterial community in uninoculated and inoculated field-grown sugarcane (Saccharum sp.)
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DOI：
10.1007/s11104-011-0812-0
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发表时间：
2012-07-01
期刊：
影响因子：
4.9
通讯作者：
Hartmann, Anton
中科院分区：
文献类型：
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作者：
Fischer, Doreen;Pfitzner, Barbara;Hartmann, Anton
利用16S rRNA和nifH转录本分析鉴定甘蔗体内活性重氮菌。这将有助于更好地理解高固氮甘蔗品种生物固氮活性的基础。以甘蔗品种Rb 867515为材料，在巴西RJ州的Serop进行了田间试验，分别进行了不接种、不接种、不接种和不接种的对照试验。以EMBRAPA-CNPAB研制的五株混合菌液为接种物。在培养第3年采集根和叶鞘样本，分析16S rRNA和nifH转录本多样性。除了在葡萄糖酸乙酸杆菌中表达nifH外。和伯克霍尔德氏菌(Burkholderia spp.)，从甘蔗地上部与Ideonella/Herbasillum相关的亲缘关系类型以及缓生根瘤菌中获得了广泛多样性的nifH序列。和根瘤菌(Rhizobium sp.)在树根中发现了。这些结果得到了16S cDNA分析的证实。从接种的细菌中，在叶鞘和根中只检测到重氮营养细菌和热带细菌的nifH转录本。通过分子分析，在甘蔗根和茎中发现了已知的和尚未培养的重氮菌。在夏末收获时，鉴定出两个接种剂混合物菌株。
To identify active diazotrophs in sugarcane, 16S rRNA and nifH transcript analyses were applied. This should help to better understand the basis of the biological nitrogen fixation (BNF) activity of a high nitrogen fixing sugarcane variety. A field experiment using the sugarcane variety RB 867515 was conducted in Serop,dica, RJ, Brazil, receiving the following treatments: unfertilised and fertilised controls without inoculation, unfertilised with inoculation. The five-strain mixture developed by EMBRAPA-CNPAB was used as inoculum. Root and leaf sheath samples were harvested in the third year of cultivation to analyse the 16S rRNA and nifH transcript diversity. In addition to nifH expression from Gluconacetobacter spp. and Burkholderia spp., a wide diversity of nifH sequences from previously uncharacterised Ideonella/Herbaspirillum related phylotypes in sugarcane shoots as well as Bradyrhizobium sp. and Rhizobium sp. in roots was found. These results were confirmed using 16S cDNA analysis. From the inoculated bacteria, only nifH transcripts from G. diazotrophicus and B. tropica were detected in leaf sheaths and roots. Known as well as yet uncultivated diazotrophs were found active in sugarcane roots and stems using molecular analyses. Two strains of the inoculum mix were identified at the late summer harvest.