Piceno YM, Noble PA, Lovell CR
Department of Biological Sciences, University of South Carolina, Columbia, SC 29208, USA
Microb Ecol. 1999 Aug;38(2):157-167. doi: 10.1007/s002489900164.
Abstract Diazotroph assemblage compositions were assessed in rhizosphere sediments from the tall and short form Spartina alterniflora growth zones over an annual cycle. Sediment cores were collected for DNA extraction and nitrogenase (acetylene reduction) activity assays, and porewater samples were analyzed for several chemical parameters in March, June, September, and December 1997. These data were collected to determine if within- or between-zone differences in the diazotroph assemblage composition correlated with differences in key environmental variables or acetylene reduction activity. Acetylene reduction rates differed between zones and within a zone over an annual period. Soluble sulfide concentrations were higher in the short form S. alterniflora zone on all dates except those in June and differed within both zones on different sample dates. nifH sequences were recovered from rhizosphere sediment DNA by PCR amplification using nifH specific primers. These amplimers were analyzed using denaturing gradient gel electrophoresis (DGGE), and the resulting patterns were compared by neural network and linear discriminant analyses. Ten prominent amplimers, four of which were apparent heteroduplexes, were observed. DGGE banding profiles showed minor differences among sampling dates and between sample zones, but the overall banding pattern was remarkably consistent. This reflects overall similarity between the amplifiable diazotroph assemblages in the tall and short S. alterniflora growth zones and substantial seasonal stability in assemblage composition.http://link.springer-ny.com/link/service/journals/00248/bibs/38n2p157.html
摘要 在一年的周期内,对互花米草高株型和矮株型生长区域的根际沉积物中的固氮微生物组合成分进行了评估。采集沉积物岩芯用于DNA提取和固氮酶(乙炔还原)活性测定,并于1997年3月、6月、9月和12月分析孔隙水样品中的几个化学参数。收集这些数据是为了确定固氮微生物组合成分在区域内或区域间的差异是否与关键环境变量或乙炔还原活性的差异相关。乙炔还原率在不同区域之间以及在一个区域内的一年时间里有所不同。除了6月的样品外,矮株型互花米草区域的可溶性硫化物浓度在所有日期都较高,并且在不同采样日期的两个区域内也有所不同。通过使用nifH特异性引物进行PCR扩增,从根际沉积物DNA中回收nifH序列。使用变性梯度凝胶电泳(DGGE)对这些扩增子进行分析,并通过神经网络和线性判别分析比较所得图谱。观察到10个突出的扩增子,其中4个明显是异源双链体。DGGE条带图谱显示采样日期之间和样品区域之间存在微小差异,但总体条带模式非常一致。这反映了互花米草高株型和矮株型生长区域中可扩增固氮微生物组合之间的总体相似性以及组合成分的显著季节稳定性。http://link.springer-ny.com/link/service/journals/00248/bibs/38n2p157.html