Vijverhof Laboratory, Limnological Institute, 3631 AC Nieuwersluis, The Netherlands.
Appl Environ Microbiol. 1989 Jul;55(7):1787-95. doi: 10.1128/aem.55.7.1787-1795.1989.
In stratified Lake Vechten, The Netherlands, protozoan grazing was estimated on the basis of uptake of fluorescently labeled bacteria and compared with bacterial production estimated on the basis of thymidine incorporation. By using a grazer-free mixed bacterial population from the lake in continuous culture, an empirical relationship between cell production and thymidine incorporation was established. Thymidine incorporation into total cold-trichloroacetic-acid-insoluble macromolecules yielded a relatively constant empirical conversion factor of ca. 10 (range, 0.38 x 10 to 1.42 x 10) bacteria mol of thymidine at specific growth rates (mu) ranging from 0.007 to 0.116 h. Although thymidine incorporation has been assumed to measure DNA synthesis thymidine incorporation appeared to underestimate the independently measured bacterial DNA synthesis by at least 1.5- to 13-fold, even if all incorporated label was assumed to be in DNA. However, incorporation into DNA was found to be insignificant as measured by conventional acid-base hydrolysis. Methodological problems of the thymidine technique are discussed. Like the cultures, Lake Vechten bacteria showed considerable thymidine incorporation into total macromolecules, but no significant incorporation into DNA was found by acid-base hydrolysis. This applied not only to the low-oxygen hypo- and metalimnion but also to the aerobic epilimnion. Thus, the established empirical conversion factor for thymidine incorporation into total macromolecules was used to estimate bacterial production. Maximum production rates (141 x 10 bacteria liter h; mu, 0.012 h) were found in the metalimnion and were 1 order of magnitude higher than in the epi- and hypolimnion. In all three strata, the estimated bacterial production was roughly balanced by the estimated protozoan grazing. Heterotrophic nanoflagellates were the major consumers of the bacterial production and showed maximum numbers (up to 40 x 10 heterotrophic nanoflagellates liter) in the microaerobic metalimnion.
在荷兰维特恩分层湖中,根据对荧光标记细菌的吸收来估计原生动物的摄食,并与根据胸苷掺入估计的细菌生产力进行比较。通过使用来自湖泊的无掠食者混合细菌群体进行连续培养,建立了细胞产量与胸苷掺入之间的经验关系。胸苷掺入到总冷三氯乙酸不可溶大分子中,在特定生长率(μ)范围从 0.007 到 0.116 h 时,产生约 10(范围从 0.38×10 到 1.42×10)个细菌 mol 的胸苷的相对恒定的经验转换因子。尽管胸苷掺入被认为可以测量 DNA 合成,但即使假设所有掺入的标记都在 DNA 中,胸苷掺入也至少低估了独立测量的细菌 DNA 合成的 1.5 到 13 倍。然而,通过常规酸碱水解发现,掺入 DNA 的量微不足道。讨论了胸苷技术的方法学问题。与培养物一样,维特恩湖的细菌在总大分子中有相当大的胸苷掺入,但通过酸碱水解未发现明显的 DNA 掺入。这不仅适用于低氧缺氧层和金属层,也适用于好氧上涌层。因此,用于估计细菌生产力的胸苷掺入到总大分子中的已建立的经验转换因子被使用。在金属层中发现了最大的生产力速率(141×10 细菌升小时;μ,0.012 小时),比在上涌层和缺氧层中高 1 个数量级。在所有三个层中,估计的细菌生产力大致与估计的原生动物摄食相平衡。异养微型鞭毛虫是细菌生产力的主要消费者,在微需氧金属层中数量最多(高达 40×10 异养微型鞭毛虫升)。