Spatial and Seasonal Variations of Microbial Communities in Dripping Water from the Heshang Cave, Central China
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摘要: 为了考察现代微生物群落对环境因子的响应关系及其在古气候重建中的应用潜力, 利用微生物学传统的培养计数、荧光染色总菌计数和BIOLOG的ECO微平板等技术对湖北省清江和尚洞内不同地点的滴水由内至外进行了空间和季节性的动态监测, 4个滴水点由内向外编号依次为1D、3D、4D和5D.结果证实湖北清江和尚洞洞穴滴水中存在一定数量的微生物.不同样点微生物的代谢活性不同, 1D样品中的微生物群落代谢活性最高, 3D表现出与1D相似的代谢特征, 但平均颜色变化率(AWCD: 微生物对碳源的利用程度, 颜色变化率越高, 表明微生物代谢越旺盛)较1D小, 4D和5D样品在接种24h后平均颜色变化率(AWCD)基本保持不变.对微生物代谢较为旺盛的1D、3D两个样品中不同时间内微生物群落的多样性指数进行分析, 表明洞穴滴水中微生物多样性相对稳定, 随采样季节的变化不大.相对于3D、1D的4个不同季节(采样时间)微生物群落对碳源种类的利用上存在一定差异.1D水样在2006秋季(2006年11月份)、2007春季(2007年5月)和2009年夏季(2009年7月份)微生物群落利用碳源种类上比较接近, 而在2008年初秋(2008年9月)碳源利用种类与前三者的差异明显.与1D相比, 3D的4个不同采样时间的滴水微生物群落在主成分1和主成分2的得分系数比较接近, 群落碳源代谢功能多样性差别不大.这种变化可能反映了不同季节滴水中可溶有机质成分(DOC)的变化及微生物群落成分的变化, 暗示了微生物群落结构变化在指示气候变化应用中的巨大潜力.但微生物群落多样性与代谢功能多样性与气候因子之间的关系尚需深入研究.Abstract: Dripping water was seasonally collected from several sites (denoted as site 1D, 3D, 4D and 5D toward the cave entrance respectively) inside the oligotrophic Heshang cave, Central China, to detect the metabolic activities of microbial communities in response to rainfall and temperature. Viable counting method, the DAPI staining technique as well as Biolog Eco microplates were exploited for microbial counting and metabolic changes of microbial communities. Results confirm the existence of microbes in the dripping water though the microbial biomass is as low as that in the deep sea water column. The metabolic activities of microbial communities varied with sites as indicated by the average well color development (AWCD) values in Biolog ECO microplates. Microbes in site 1D showed the strongest metabolic activities followed by those in site 3D. Microbial communities in the sites near the cave entrance, such as site 4D and site 5D, however, displayed the weakest metabolic activities as shown by the invariant AWCD values after 24-hour incubation. Similar microbial diversity indices were observed between different sites such as site 1D and site 3D, and even among different seasons. This indicates the presence of a fairly stable microbial diversity in the dripping water, probably due to the oligotrophic condition in the cave. However, the carbon utilization pattern of microbial communities in site 1D differed greatly among seasons; the pattern in Sept. 2008 is distinguishable from those in other sampling periods. The seasonal variation in carbon utilization might arises from the seasonal changes of dissolved organic matter or the changes in the flow rate of dripping water. Noticeably, such kind of seasonal variation in the carbon utilization was not reproduced in site 3D. Yet we need further investigations to determine the relationship between the changes in microbial community and the climate factors such as temperature and rainfall.
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Key words:
- karst cave /
- climate change /
- microbiology /
- community diversity
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图 1 和尚洞洞穴滴水微生物群落利用碳源的平均颜色变化率随培养时间的变化
a.2006年11月不同滴点微生物群落碳源代谢特征；b.不同季节1D滴水微生物群落碳源代谢特征；c.不同季节3D滴水微生物群落碳源代谢特征
Fig. 1. Changes of average well color development with incubation times of the dripping waters in Heshang cave
图 2 和尚洞滴水微生物群落碳源代谢主成分分析
a.不同季节的1D滴水微生物群落；b.不同季节的3D滴水微生物群落
Fig. 2. Principle analysis of carbon metabolism of microbial communities in dripping waters in Heshang cave
图 3 和尚洞1D和3D不同季节滴水中的微生物群落多样性指数
Fig. 3. Diversity indices of microbial communities in 1D and 3D in different seasons in Heshang cave
表 1 湖北清江和尚洞滴水微生物计数结果
Table 1. Total microbial counts in dripping waters of Heshang cave, Hubei
样品号 琼脂平板计数(CFU·mL-1) 荧光染色计数(Cells·mL-1) 可培养比率(%) 2008.09 1D 4.85×102 7.668×104 0.63 2008.09 3D 2.18×102 2.628×104 0.83 
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