It has long been believed that soil microbial activity in tropical forest ecosystems is limited by phosphorus (P) rather than nitrogen (N) availability. In this study, we reviewed the methods used to determine the limiting nutrients and evaluated the validity of the widespread P‐limitation hypothesis in tropical forest soils. The most commonly used analysis method entails testing whether fertilization increased microbial biomass or soil respiration. Fertilization using microbial biomass as an indicator was not a satisfactory method because standing microbial biomass does not always signal microbial activity. An increase in soil respiration after fertilization was also an insufficient indicator because the negative response shown by organic matter decomposition to nutrient addition can also be driven by nutrient shortage (nutrient mining). Nutrient amendment can also shift microbial communities towards more copiotrophic organisms, which may exhibit lower microbial respiration rates. We suggest that P addition may accelerate soil organic matter decomposition compared with N, which is independent of the nutrient limitation of soil microbial activity. The negative response of organic matter decomposition to N addition through nutrient mining is driven by N shortage, which is less likely to occur with P. N addition inhibits microbial activity via chemical reactions, whereas P addition may stimulate activity through replacement by C bound to sorption sites in the soil, improving C availability. Thus, the P‐limitation hypothesis must be reconsidered because of the contrasting effects of exogenous N and P addition on soil microbial activity, which could lead to misdetection of P limitation on soil microbial activity in tropical forest ecosystems. We recommend caution in applying the statement that soil microbial activity in tropical forest ecosystems is limited by P until a novel method is established to accurately determine the nutrients limiting soil microbial activity at the ecosystem level. We proposed alternative ways to describe nutrient shortage for soil microbes. Aplain language summaryis available for this article.

Previous meta-analyses revealed that the ratio of activities of carbon (C)-acquiring enzyme to nitrogen (N)-acquiring enzymes in tropical forest ecosystems was nearly identical to those in other ecosystems, despite of the N-rich condition in tropical forests. This could be explained by microbes in tropical forest soils, which require a large amount of N to produce N-rich acid phosphatase (AP) for catalyzation of the organic form of phosphorus (P) and compensation for poor P availability in soils. Based on this idea, we hypothesized that experimental P fertilization would reduce the allocation to N-acquiring enzymes compared with that of C-acquiring enzymes, i.e. that it would increase the ratios of activities of β-1,4-glucosidase (BG) to β-1,4-acetylglucosaminidase (NAG) and leucine aminopeptidase (LAP). We tested this hypothesis using an experimental fertilization site with factorial N (100 kg ha⁻¹ yr⁻¹) and P (50 kg ha⁻¹ yr⁻¹) addition in a primary tropical lowland forest in Bornean Malaysia, where our earlier work demonstrated that P fertilization reduced AP activity. Contrary to our hypothesis, the BG:NAG and BG:(NAG + LAP) ratios were not altered by either N or P fertilizations. This result indicated that AP production was not a reason for the maintenance of a relatively high investment in N-acquiring enzyme at our study site. Rather, NAG and LAP production was likely driven by C acquisition, rather than N acquisition, as the target substrates contained C as well as N. This idea was supported by the fact that neither the BG:NAG ratio nor the BG:(NAG + LAP) ratio was elevated by N addition. We propose that the ratios of activities of BG to NAG and LAP do not necessarily indicate the ratio of C:N acquisition, at least in our N-rich tropical forest ecosystem.

The tea bag method provides a replicable and standardized method to study the effect of environmental variables on the decomposition of standard litter, which enables comparison of organic matter decomposition rates on a large scale. However, it remains uncertain whether tea bag decomposition in response to wetness is representative of that of local litters. We performed incubation experiments to examine whether the effect of soil water on tea bag decomposition becomes inhibitory at higher water contents, as is the case in local leaf litters. In addition, we performed field studies in a mixed forest and cedar plantation in Japan to compare two litter bag mesh sizes: 0.25-mm mesh, the size previously used by a major manufacturer of tea bags (Lipton), and nonwoven bags with mesh sizes finer than 0.25 mm, which are currently produced by Lipton. Both green tea and rooibos tea exhibited higher decomposition rates at higher water contents, but decomposition was inhibited at the highest water content; this was in contrast to our hypothesis based on a field observation but consistent with conceptual models of local litters. The nonwoven tea bags did not show lower decomposition rates, despite the finer mesh size. Rather, the nonwoven rooibos tea bags exhibited slightly higher decomposition rates than the 0.25-mm mesh bags in the cedar plantation, possibly due to a greater abundance of microorganisms that decompose litters in the nonwoven bags, due to the decrease in predation by mesofauna. Our findings provide essential information for future studies of tea bag decomposition.