Department of Oceanography, University of Hawaii, Honolulu, Hawaii 96822.
Appl Environ Microbiol. 1979 Nov;38(5):850-60. doi: 10.1128/aem.38.5.850-860.1979.
A relatively simple and extremely sensitive technique for measuring rates of stable ribonucleic acid (RNA) synthesis was devised and applied to bacterial cultures and seawater samples. The procedure is based upon the uptake and incorporation of exogenous radiolabeled adenine into cellular RNA. To calculate absolute rates of synthesis, measurements of the specific radioactivity of the intracellular adenosine 5'-triphosphate pools (precursor to RNA) and of the total amount of radioactivity incorporated into stable cellular RNA per unit time are required. Since the rate of RNA synthesis is positively correlated with growth rate, measurements of RNA synthesis should be extremely useful for estimating and comparing the productivities of microbial assemblages in nature. Adenosine 5'-triphosphate, adenylate energy charge, and rates of stable RNA synthesis have been measured at a station located in the Columbian Basin of the Caribbean Sea. A subsurface peak in RNA synthesis (and therefore growth) was located within the dissolved oxygen minimum zone (450 m), suggesting in situ microbiological utilization of dissolved molecular oxygen. Calculations of the specific rates of RNA synthesis (i.e., RNA synthesis per unit of biomass) revealed that the middepth maximum corresponded to the highest specific rate of growth (420 pmol of adenine incorporated into RNA.day) of all depths sampled, including the euphotic zone. The existence of an intermediate depth zone of active microbial growth may be an important site for nutrient regeneration and may serve as a source of reduced carbon for mesopelagic and deep sea environments.
我们设计并应用了一种相对简单且极其灵敏的技术来测量稳定核糖核酸(RNA)的合成速率,该技术可用于细菌培养物和海水样本。该方法基于外源性放射性标记腺嘌呤的摄取和掺入细胞 RNA。为了计算合成的绝对速率,需要测量细胞内腺苷 5'-三磷酸池(RNA 的前体)的比放射性和单位时间内稳定掺入细胞 RNA 的放射性总量。由于 RNA 合成速率与生长速率呈正相关,因此测量 RNA 合成速率对于估计和比较自然界中微生物群落的生产力应该非常有用。在加勒比海的哥伦比亚盆地的一个站位测量了腺苷 5'-三磷酸、腺苷酸能荷和稳定 RNA 合成的速率。在溶解氧最小值区(450 m)内发现了 RNA 合成(因此是生长)的次表层峰值,这表明原位微生物利用了溶解的分子氧。RNA 合成的比速率(即单位生物量的 RNA 合成)的计算结果表明,中深最大处对应于所有采样深度中最高的特定生长率(420 pmol 腺嘌呤掺入 RNA.day),包括透光带。中间深度活跃微生物生长区的存在可能是营养物质再生的重要场所,并可能成为中上层和深海环境中还原碳的来源。