Labile organic carbon pools and enzyme activities of Pinus massoniana plantation soil as affected by understory vegetation removal and thinning.

The effects of forest management on carbon (C) sequestration are poorly understood, particularly in the Three Gorges Reservoir area. We aimed to identify the effects of forest management on C sequestration in Pinus massoniana plantations. An intact control forest (CK), a site undergoing regular shrub cutting with the simultaneous removal of residues (SC), a site under low-intensity thinning (LIT), and a site under high-intensity thinning (HIT) were compared for soil labile organic carbon (LOC), related enzyme activities, and soil characteristics. Soil organic carbon (SOC) significantly decreased in the HIT treatment as compared with that in the CK treatment. Soil EOC, DOC, MBC contents in treated plots were higher than those in the CK treatment; particularly, the HIT treatment significantly increased those values in 0-10 cm layer. Thinning resulted in a decrease in cellulase and amylase activities, but an increase in invertase activity. In addition, the SOC content was significantly correlated with four enzymes activities and LOC components, which suggested that the soil LOC components and enzymes activities were sensitive to the changes of SOC. Our results suggest that high-intensity thinning treatment in Pinus massoniana plantation could significantly decrease the SOC content and lead to an increase of LOC components.

Introduction

The forest ecosystem accounts for approximately 73% of the terrestrial soil papan class="Chemical">n class="Chemical">carbon (C) pool, which is anpan> important part of the global terrestrial ecosystem[1]. Thus, the forest ecosystem plays a crucial role in the C cycle[2]. A major focus of forest manpan>agemenpan>t is to promote the incremenpan>t of C pool[3-5]. In soil, according to their meanpan> residenpan>ce times, soil pan> class="Chemical">organic carbon (SOC) can be divided into recalcitrant and labile components. Management practices have little effect on recalcitrant components because of their longer turnover time in soil[6], while soil labile organic carbon (LOC) fractions are more responsive to changes in forest management strategies. Although soil LOC fractions make up a relatively small part of SOC[7-9], they can serve as indicators of minor changes in SOC.

Responses of soil LOC compopan class="Chemical">nents are often indicated by easily oxidised organic carbon (EOC), dissolved pan> class="Chemical">organic carbon (DOC), and microbial biomass carbon (MBC)[6,10]. Previous studies have shown that EOC, DOC, and MBC contents affect C sequestration capacity of soil and the emission of greenhouse gases[11] thus indicating that they are important sources of C that are released from the soil to the atmosphere and aid in the decomposition of recalcitrant C[12]. Thus, soil LOC fractions in forests are imported for maintaining balance in the soil C pool under different forest management strategies. Moreover, the activities of enzymes related to the soil C cycle (e.g., invertase, amylase, and cellulase) participate in the SOC decomposition and indicate the status of the available C resources. Therefore, these enzyme activities can contribute to our understanding of the variations in SOC in response to forest management[13,14].

Thinpan class="Chemical">ning is a common strategy of forest management used in plantations forests, and it is a tool for controlling the species composition[15]. Thinning commonly decreases SOC by reducing litter inputs into soil anpan>d possibly accelerating decompositionpan> rates owing to chanpan>ge in microclimate[3,12] anpan>d their chanpan>ge degree of pan> class="Chemical">SOC among different soil layers were varied[12]. Understory vegetation can modulate SOC by affecting the soil characteristics and changing the organic inputs and the leaching of dissolved organic matter[16]. Different forest management strategies can change soil temperature and moisture, and the composition of aboveground vegetation, thereby influencing C and nutrient cycles in forests[17-19]. For example, whole-tree harvesting has the most significant effect on the soil C pool, because it causes direct damage to vegetation via SOC release, increased soil temperature, and accelerated erosion[20,21]. Accumulating evidence shows that for most tree species, the effect of forest thinning on SOC dynamics is complex, and that the intensity and method of forest management affect the degree to which microclimate and residual vegetation composition are affected, thereby affecting SOC sequestration[3,22]. However, studies on forest C storage under different forest management strategies have produced contradictory conclusions[23,24]. It has been reported that decreasing tree density in stands decreases total forest C stores[25,26], whereas others have found that this action can maintain or increase live tree C due to the increased growth rate of trees grown at lower densities[27], especially in the case of long-term responses to thinning[28]. Short-term studies have revealed that thinning consistently decreases aboveground C[26,29], indicating that low densities of small trees do not fully offset the loss of C[30]. Thus, it is still unclear whether forest management strategies are compatible with the purpose of increasing forest C storage for climate change mitigation. In addition, exploring the effects and mechanisms affecting forest management strategies on SOC sequestration is a key research area in both forestry and C cycling science, while relatively less attention was paid to the effect of short-term forest management activities on soil LOC fractions[31].

Plantatiopan class="Chemical">ns are a key component of global forest resources, and play an important role in sustainable forest management[32-34]. In China, pure plantations forests constitute approximately 80% of all plantations[16]. Pinus massoniana is the main tree species used for afforestationpan> in South China. It plays a major role in providing forest resources anpan>d ecological services[35], anpan>d it covers the largest area (67.83 × 104 ha) in the Three Gorges Reservoir area located in subtropical China[36]. However, due to anpan>thropogenpan>ic effects anpan>d the complex terrain in this regionpan>, the soil C conpan>tent is relatively low[37]. Thus, in recenpan>t years, recreating forest structures anpan>d optimizing the use of soil by forest manpan>agemenpan>t activities were a major focus of the sustainable manpan>agemenpan>t of pan> class="Species">Pinus massoniana aimed at maintaining ecosystem sustainability and soil productivity[12].

In this study, the objective was to assess the effect of forest mapan class="Chemical">nagement in term of SOC, LOC fractionpan>s, anpan>d activities of related enpan>zyme at three minpan>eral soil layers (0–10 cm, 10–20 cm, 20–30 cm). The aim of the project was to investigate (1) how differenpan>t forest manpan>agemenpan>t treatmenpan>ts (0%, 15% anpan>d 70% stem thinning anpan>d understory vegetationpan> removal) influenpan>ce the conpan>tenpan>ts of pan> class="Chemical">SOC, soil LOC fractions (i.e. DOC, MBC, and EOC), and related enzyme activities (i.e. cellulase, amylase, invertase, and catalase) and (2) whether relationships exist among soil LOC fractions, enzyme activities and other soil characteristics (soil pH and fertility characteristics). Our hypotheses are that 1) thinning and understory vegetation removal treatment will decrease the SOC, enzyme activities and increase LOC pools, and 2) the soil LOC fractions are linked to enzyme activities and partial soil characteristics.

Results

Soil chemical properties

Soil total nitrogenpan> (TN), total phosphorous (TP), available potassium (AK), NH4+–N, and NO3−–N contents decreased with increasing soil depth while soil pH increased with increasing depth, although significant difference were only observed in the soil TN content and pH among the soil layers (p < 0.05). Besides this, in the 0–10 cm soil layer, TN, total potassium (TK), AK and NO3−–N contents were significantly lower in the three treated plots than those in the control plots (p < 0.05). In the 10–20 cm soil layer, AK and NO3−–N contents in the three treated plots were significantly lower than those in the control plots (p < 0.05). In addition, TN content in the SC treatment plots was greater than those in the thinning (LIT, HIT) plots in 0–10 cm and 10–20 cm soil layer (p < 0.05). In the 20–30 cm soil layer, NH4+–N and NO3−–N contents in three treated plots were significantly lower than that in the control plots (p < 0.05) (Table 1).

Table 1

Soil chemical properties at three soil depths in the four forest management treatments (mean value ± standard error; n = 3). Significant differences among different soil layers subjected to the same treatments are identified with A, B, and C (p < 0.05). Significant differences among different treatments of the same soil layer are identified with a, b, c, and d (p < 0.05), based on the analysis of variance.

Treat- ments	Soil depth (cm)	Soil pH	TN	TP	TK	AP	AK	NO3−–N	NH4+–N
CK	0–10	5.85 ± 0.02 Aa	1.65 ± 0.01 Aa	0.21 ± 0.01 Aa	17.05 ± 0.12 Aa	0.84 ± 0.10 a	184.68 ± 2.19 Aa	17.79 ± 0.89 Aa	43.00 ± 3.51 Aa
	10–20	5.92 ± 0.06 Ba	1.15 ± 0.01 Ba	0.18 ± 0.01 Ba	16.76 ± 0.13 Ba	0.99 ± 0.17 a	145.83 ± 2.80 Ba	11.26 ± 0.99 Ba	37.34 ± 3.48 Ba
	20–30	6.07 ± 0.05 Ca	0.97 ± 0.01 Ca	0.17 ± 0.01 Ba	17.13 ± 0.10 Ba	0.98 ± 0.17 a	136.58 ± 0.47 Ca	5.40 ± 0.0.37 Ca	24.66 ± 0.56 Ca
SC	0–10	6.02 ± 0.05 Aa	1.57 ± 0.02 Ab	0.20 ± 0.01 Aa	18.21 ± 0.15 Ab	1.23 ± 0.17Aa	146.45 ± 2.04 Ab	11.65 ± 0.22 Ab	45.66 ± 0.69 Aa
	10–20	6.16 ± 0.03 Ba	1.16 ± 0.01 Ba	0.19 ± 0.01 Ba	18.08 ± 0.25 Bb	2.35 ± 0.29 Bb	126.18 ± 1.28 Bb	4.48 ± 0.14 Bb	18.96 ± 0.23 Bb
	20–30	6.33 ± 0.03 Ca	0.90 ± 0.01 Cb	0.18 ± 0.01 Ca	19.52 ± 0.19 Ca	1.57 ± 0.4 ABab	110.43 ± 2.47 Cb	2.75 ± 0.34 Cb	12.76 ± 1.28 Cb
LIT	0–10	6.17 ± 0.02 Aa	1.37 ± 0.04 Ac	0.19 ± 0.01 Aa	16.54 ± 0.21 Ac	2.11 ± 0.39 b	140.60 ± 3.29 Acb	9.73 ± 0.42 A c	50.69 ± 2.91 Ab
	10–20	6.25 ± 0.03 Ba	1.04 ± 0.02 Bb	0.17 ± 0.01 Ba	17.14 ± 0.08 Ba	1.46 ± 0.92 ac	127.40 ± 2.50 Bb	2.57 ± 0.10 Bc	14.79 ± 0.47 Bc
	20–30	6.48 ± 0.02 Ca	0.85 ± 0.0 1 Cc	0.17 ± 0.01 Ba	17.70 ± 0.24 Aa	2.51 ± 0.61 b	126.25 ± 3.29 Aa	2.45 ± 0.13 Bb	10.99 ± 0.28 Cb
HIT	0–10	5.97 ± 0.05 Aa	1.49 ± 0.01 Ad	0.19 ± 0.01 Aa	16.43 ± 0.16 Acd	0.82 ± 0.70 Aa	130.78 ± 0.52 Ac	5.35 ± 0.19 Ad	35.79 ± 3.29 Ac
	10–20	6.07 ± 0.03 Ba	1.09 ± 0.02 Bc	0.17 ± 0.01 Ba	15.83 ± 0.34 Bc	1.09 ± 0.10 ABad	82.90 ± 1.60 Bc	4.90 ± 0.58 ABb	34.51 ± 2.41 Aa
	20–30	6.20 ± 0.04 Ca	0.96 ± 0.01 Ca	0.18 ± 0.01 Ca	16.58 ± 0.05 Ca	1.91 ± 0.90 Bab	71.68 ± 1.57 Cc	3.50 ± 1.56 Bb	29.25 ± 1.50 Bc

Soil chemical properties at three soil depths in the four forest mapan class="Chemical">nagement treatments (mean value ± standard error; n = 3). Significant differences among different soil layers subjected to the same treatments are identified with A, B, and C (p < 0.05). Significant differences among different treatments of the same soil layer are identified with a, b, c, and d (p < 0.05), based on the analysis of variance.

LOC fraction

SOCpan>, EOC, DOC, and MBC decreased with increasing soil depth, and significant difference were only observed in the SOC and DOC contents among the three soil layers (p < 0.05). The EOC content in the 0–10 cm layer was significantly higher than that in the other soil layers (p < 0.05). In the 0–10 cm soil layer, SOC content under the LIT and HIT treatment were significantly lower than those in the CK and SC plots, but in 0–30 cm soil layer, significant difference between the HIT and CK treatment was only observed (p < 0.05). The contents of DOC under the SC and HIT treatment were significantly higher than those in CK and LIT treatment in 0–10 cm soil layer (p < 0.05), and the corresponding values in the 10–20 cm layer were significantly higher than those in the CK (p < 0.05). The content of EOC in the 0–10 cm layer was significantly higher in the HIT treatment than that in other three treated plots (p < 0.05). In addition, the content of MBC in the three treated plots were higher than those in the CK, and, in particular, there was only a significant difference between LIT and CK treatment observed in three soil layers (p < 0.05) (Fig. 1).

Figure 1

Soil LOC fractions in the four forest management treatments. The three columns in each treatment represent the quantities in soil LOC content at different soil depths. Significant differences among different soil layers subjected to the same treatments are identified with A, B, and C (p < 0.05). Significant differences among different treatments of the same soil layer are identified with a, b, c, and d (p < 0.05), based on the analysis of variance. Values are means ± standard error (n = 3).

Soil LOC fractions ipan class="Chemical">n the four forest management treatments. The three columns in each treatment represent the quantities in soil LOC content at different soil depths. Significant differences among different soil layers subjected to the same treatments are identified with A, B, and C (p < 0.05). Significant differences among different treatments of the same soil layer are identified with a, b, c, and d (p < 0.05), based on the analysis of variance. Values are means ± standard error (n = 3).

Soil enzyme activity

Soil enzyme activity decreased with ipan class="Chemical">ncreasing soil depth from the overall, which was consistent with the trend of vertical change in LOC content (Fig. 2). Soil cellulase enzyme activities in the 0–10 cm soil layer were higher than those in the other two soil layers (p < 0.05), and the activities of cellulase and invertase in the 10–20 cm soil layer were significantly higher than those in the 20–30 cm layer (p < 0.05).

Figure 2

Soil enzymes in the four forest management treatments. The three columns in each treatment represent the quantities of four soil enzymes at different soil depths. Significant differences among different soil layers subjected to the same treatments are identified with A, B, and C (p < 0.05). Significant differences among different treatments of the same soil layer are identified with a, b, c, and d (p < 0.05), based on the analysis of variance. Values are means ± standard error (n = 3).

Soil enzymes ipan class="Chemical">n the four forest management treatments. The three columns in each treatment represent the quantities of four soil enzymes at different soil depths. Significant differences among different soil layers subjected to the same treatments are identified with A, B, and C (p < 0.05). Significant differences among different treatments of the same soil layer are identified with a, b, c, and d (p < 0.05), based on the analysis of variance. Values are means ± standard error (n = 3).

Comparison with CK revealed that, the LIT apan class="Chemical">nd HIT treatmenpan>t resulted inpan> lower levels of cellulase, amylase, and catalase activity, whereas invertase activity increased. Cellulase activity in the 0–10 soil layer in the CK plots was significanpan>tly higher thanpan> that in the SC anpan>d thinning plots (p < 0.05). Amylase activity in the 0–10 anpan>d 10–20 cm soil layers in the CK plots were significanpan>tly higher thanpan> those in the LIT anpan>d pan> class="Disease">HIT plots (p < 0.05). The differences of soil invertase activity between the LIT, HIT, and CK plot the in 10–20 cm and 20–30 cm layers were significant (p < 0.05).

Relationships between soil LOC fractions and soil enzyme activities

SOCpan> contenpan>t was significanpan>tly positively correlated with the conpan>tenpan>t of pan> class="Chemical">DOC, MBC, and EOC (r = 0.898, p < 0.01; r = 0.704, p < 0.01; r = 0.466, p < 0.01, respectively), as well as with the activities of cellulase, amylase, catalase, and invertase (r = 0.930, p < 0.01; r = 0.311, p < 0.05; r = 0.725, p < 0.01; r = 0.570, p < 0.05, respectively) (Table 2). The content of DOC, MBC, and EOC were significantly positively correlated with cellulase, catalase, and invertase activity and TN content (p < 0.01), but negatively correlated with TK and available phosphorous (AP) content, although this negative correlation was not significant). There was a significant positive correlation between DOC content and AK, NH4+–N, and NO3−–N content (r = 0.465, p < 0.01; r = 0.604, p < 0.01; r = 0.495, p < 0.01, respectively) and a negative correlation with pH (r = −0.308, p < 0.05). MBC content showed a significant positive correlation with AK and NH4+–N content (r = 0.337, p < 0.05 and r = 0.413, p < 0.01, respectively). There was significant correlation among DOC, MBC, and EOC, indicating that the components of LOC were closely related to each other. The activities of the four enzymes in this study were significantly correlated with the content of TN, NO3−–N, NH4+–N, and TP in the soil.

Table 2

Correlation of soil LOC content or enzyme activities with soil characteristics (n = 3). Significant correlations are indicated by *p < 0.05 or **p < 0.01 based on Pearson’s correlation analysis.

	SOC	DOC	MBC	ROC	Cellulase	Amylase	Catalase	Invertase	pH	TN	TP	TK	AP	AK	NH4+–N
DOC	0.898**
MBC	0.704**	0.727**
ROC	0.466**	0.460**	0.381**					+
Cellulase	0.930**	0.866**	0.573**	0.434**
Amylase	0.311*	0.088	−0.073	0.022	0.380**
Catalase	0.725**	0.687**	0.619**	0.406**	0.710**	0.131
Invertase	0.570*	0.541**	0.518**	0.384**	0.491**	0.016	0.558**
pH	−0.359*	−0.308*	−0.019	−0.140	−0.464**	−0.421**	−0.366*	0.164
TN	0.850**	0.861**	0.611**	0.395**	0.884**	0.311*	0.665**	0. 524*	−0.597**
TP	0.679**	0.679**	0.423**	0.327*	0.721*	0.334*	0.423**	0.44*	−0.430**	0.862**
TK	−0.130	−0.156	−0.026	−0.088	−0.227	−0.105	−0.328*	−0.335*	0.572**	−0.179	0.171
AP	−0.201	−0.211	−0.035	−0.176	−0.254	−0.200	−0.157	0.199	0.520**	−0.329*	−0.208	0.268
AK	0.635**	0.465**	0.337*	0.249	0.635**	0.588*	0.274	0.165	−0.287	0.599**	0.609**	0.131	−0.202
NH4+–N	0.575**	0.604**	0.413**	0.032	0.628**	0.508*	0.531**	0.401*	−0.741**	0.785**	0.547**	−0.491**	−0.294*	0.313*
NO3−–N	0.599**	0.495**	0.244	0.040	0.679**	0.597**	0.416**	−0.540*	−0.738**	0.793**	0.728**	−0.174	−0.293*	0.715**	0.771**

Correlation of soil LOC copan class="Chemical">ntent or enzyme activities with soil characteristics (n = 3). Significant correlations are indicated by *p < 0.05 or **p < 0.01 based on Pearson’s correlation analysis.

Discussion

The contepan class="Chemical">nt of soil TN, pan> class="Chemical">TP, AK, NH4+–N and NO3−–N decreased with increasing soil depth, and pH exhibited the opposite trend, which is consistent with previous findings[38-40]. According to Zhang et al.[39], SOC and TN of a chestnut forest decreased after shrub cutting. In this study, contents of TN, AK, and NO3−–N in the 0–10, 10–20, and 20–30 cm soil layers were reduced in the LIT and HIT treatment. This might be due to a reduction in nutrient elements such as N, P, and calcium being returned to the soil through the litter, during to the thinning proces[41]. Moreover, we did not find any significant effect of treatment on soil pH and TP of all the soil layers, which were consistent with the findings of many studies[42-44]. This might be due to the positive and negative effects of the treatments on soil nutrients[42]. It is also possible that the effects of treatments on these properties will be manifested in the long term but not observed in the short period of this study[45].

In this study, papan class="Chemical">n class="Chemical">SOC conpan>tent of soil (0–30 cm) in the CK, SC, LIT, anpan>d pan> class="Disease">HIT treatment were 53.01, 53.84, 53.31, and 48.70 g kg−1, respectively, suggesting that HIT treatment reduced the SOC content remarkably, which was partially consistent with the hypothesis. Similar results were found by Chen et al.[12] and Achat et al.[46]. The decrease in SOC content was mainly caused by the fact that, after the thinning, substrate inputs to the soil were reduced[3]. The microclimate induced an increase in the rate of SOC decomposition following decreased canopy closure and reduced SOC content[47]. In addition, the partial removal of tree canopies caused acceleration SOC leaching losses[12]. Moreover, we found that the response of SOC to thinning and understory vegetation removal was significantly different among three different soil layers because of the SOC in deeper soil layer with a longer residence time is less sensitive to disturbances[48]. These results may be due to the different compositions of plant species in different treatment plots wherein their roots reach different soil depths to control C sequestration and decomposition[49].

SOCpan>, DOC, MBC, and EOC content decreased with increasing soil depth, which is consistent with previous findings[8,50-52]. This may be related to the spatial distribution of root residual input and decomposition[52,53]. Although little is known about the composition of soil active C, the method has shown that it is more sensitive to soil management strategies than SOC is, and more closely related to soil biological properties[7,54-56]. According to Chatterjee et al.[57] and Diochon et al.[58], forest thinning causes in the return of a large amount of residual organic matter to the surface, accompanied by changes in light conditions, which can lead to significant changes in LOC mineralisation. In our work, the contents of soil EOC, MBC, and DOC in thinning treatments were higher than those in the CK plots was consistent with our hypothesis; particularly, the HIT treatment significantly increased these contents in the 0–10 cm layer and the effect was more marked in the upper than in the lower soil layers. The result was supported by the findings from a study on coniferous forests in Wyoming, USA[57] and those of Chen et al.[12]. However, high-intensity thinning promoted non-tree vegetation owing to the sparse forest canopy. Subsequently, their fine roots would increase the LOC input by root exudates. In addition, high-intensity thinning accelerated the decomposition of litter and residue, and the accumulation of LOC fractions, consequently the potential SOC mineralisation rate increased[12,59].

Soil enzymes participate ipan class="Chemical">n almost every transformation process of litter decomposition and play a central role in maintaining forest soil fertility by releasing plant available mineral nutrients from complex organic resources[60,61]. Soil invertase, cellulase and catalase activities decreased with increasing soil depth, in accordance with the results of Xiao et al.[52] and Chen et al.[12]. A high content of organic matter in surface soil is beneficial to the growth of microorganisms with active metabolic processes, which in turn leads to the accumulation of soil enzymes in the surface layer. In this study, the overall cellulase and amylase activities decreased after thinning, which is supported by Chen et al.[12]. Similar results were observed in the New Jersey Pine Barrens, where cellulase and phenol oxidase activities significantly decreased after one year of thinning[62]. The results were partially consistent with hypothesis of this study. These variations in enzyme activities can be due to reductions in root activity and changes in microbial composition. Furthermore, the extent of utilisation of the C and N sources for soil enzymes differs, with varying effects on soil enzyme activity under different management strategies. Li et al.[63] reported that invertase can break down some carbohydrate pan> class="Chemical">polymers to release the nutrients from organic compounds through its role in the first phases of degradation of organic compounds. During this phase, molecular size is reduced and smaller organic structures are produced, which facilitates microbial enzyme activities. We found that the thinning treatments had greater invertase activity than that in the CK treatment. A plausible explanation is that invertase activity is positively correlated with soil pH, TN, and TP, but it is negatively correlated with TK and NO3−–N.

We found that the papan class="Chemical">n class="Chemical">SOC conpan>tent was significanpan>tly correlated with the four enpan>zymes activities anpan>d soil LOC componpan>enpan>ts, which was observed in earlier studies[63,64]. The result suggested that the soil LOC componpan>enpan>ts anpan>d these enpan>zymes activities were senpan>sitive to the variationpan>s of pan> class="Chemical">SOC which was consistent with the hypothesis. Significant correlations were found among the LOC fractions and invertase, catalase, and cellulase activities in the soil, which is consistent with findings of Paz-Ferreior[65]. In addition, significant correlations were found between each component of SOC and the soil TN content in the soil, consistent with findings of Geng et al.[62]. These correlations might have arisen because the N content in the soil organic matter affects the rate of soil organic matter decomposition and consumption by microorganisms. Nitrogen-rich organic matter is easily and rapidly decomposed, transferred, and converted by microorganisms, thus increasing the SOC content in the soil. As soil enzymes directly participate in the utilisation of soil nutrients, they indirectly reflect the dynamic state of the conversion of soil nutrients. Taken together, a similar conclusion to that of Ma et al.[66] can be drawn in that enhancing soil nutrient content is the key factor for increasing the accumulation of LOC fractions.

Conclusions

This study demonstrates the distributiopan class="Chemical">n of chemical properties of soil, LOC fractions, and enzyme activities, and provides insight into their relationships in Pinus massoniana planpan>tationpan>s under differenpan>t forest manpan>agemenpan>t approaches. Our results showed that the conpan>tenpan>t of soil LOC fractionpan>s, npan> class="Chemical">TN, TP, AK, NH4+–N, NO3−–N and enzyme activities decreased with increasing soil depth in all the treatments. High-intensity thinning reduced the SOC content remarkably. Soil EOC, DOC, and MBC contents were higher in thinning treatments than those in the control; especially high-intensity thinning treatment significantly increased those contents in 0–10 cm layer. Simultaneously, thinning resulted in a decrease in cellulase and amylase activities, but an increase in invertase activity. The variations of SOC, EOC, DOC, MBC, cellulose, amylase, and invertase were more marked in the top soil layer than in the deeper soil layers. The correlation analysis showed that the soil LOC components and enzymes activities were sensitive to the variations in SOC. Our results indicate that high-intensity thinning treatments in Pinus massoniana plantation decreased the SOC content and might lead to an increase of LOC components, in which the effect was more marked in the 0–10 cm soil layer than in the deeper layers.

Materials and Methods

Study site

The experimental plots were located ipan class="Chemical">n the Jiulingtou Forest in Zigui County, Hubei Province, China, where Pinus massoniana were aerially seeded in the 1970s (Fig. 3). The soil type of the plots was haplic pan> class="Chemical">luvisol[67], and the annual mean temperature was approximately 16.9 °C with annual rainfall varying between 1000 and 1250 mm, occurring primarily from April to September[68]. Twelve 20 m × 20 m plots within Pinus massoniana monoculture plantations, laid out in an orthogonal design and separated from each other by at least 2 m, were defined in September 2013. Each plot contained four treatments, randomly assigned per row. Treatments consisted of (1) intact forest (control, CK), (2) all shrubs harvested and residual materials removed (shrub cutting, SC), (3) low-intensity thinning (LIT) with 15% of the basal area of the trees removed, and (4) high-intensity thinning (HIT) with 70% of the basal area of the trees removed. The study area was relatively steep with a northwest-facing slope of 34°. The experiment was conducted in mid-October 2013 and included a control intact forest, and thinning and shrub cutting treatments using chain saws. Only the harvested trunks were removed and the residue produced by the harvest was not cleared from the plots. Except for plots subjected to the SC treatment, the understory shrub layer was dominated by Lespedeza bicolor Turcz., Pyracantha fortuneana (Maxim.) Li, and Litsea pungens Hemsl., with an average diameter at breast height of 5.00 cm and average height of 5.60 m. Herbs in the study area were mainly Woodwardia japonica L.f., Carex tristachya Thunb., Aster ageratoides Turcz., and Parathelypteris nipponica (Franchet and Savatier) Ching.

Figure 3

Location of the study area. The study area, located in the Jiulingtou Forest Farm (30°59′N, 110°47′E), is indicated by a triangle and Zigui City is marked in green. The blue line represents the Yangtze River flowing through the Three Gorges Reservoir area (the region surrounded by the red line). The maps were created using ArcGIS 10.2 software (ESRI 2014).

Soil analysis

The twelve plots (4 treatments × 3 replicatiopan class="Chemical">ns per treatment) were divided into 24 subplots (10 m × 20 m). Six randomly-placed replicate soil samples were collected from each subplot in June 2016 at depths of 0–10, 10–20, and 20–30 cm yielding 432 (24 subplots × 6 replicates per subplot × 3 soil layers) samples, which were collected using a soil sampler 30 cm in length. These samples were placed in plastic bags and stored in a portable cooler for transport to the laboratory. The soil samples were divided into two subsamples of equal volume. One was passed through a 2-mm sieve to remove impurities of soil and stored at 4 °C before testing. This subsample was used to determine NH4+–N, NO3−–N, pan> class="Chemical">DOC, MBC, EOC, and enzyme activities (cellulase, amylase, invertase, and catalase). The other subsample was air-dried and sieved before use for the analysis of SOC and other soil properties (TN, TP, TK, AK, AP, and pH).

Sample analyses

Soil chemical analysis

SOCpan> contenpan>t was measured using dichromate oxidationpan>[69]. Soil pan> class="Chemical">TN was determined using the Kjeldahl method[70]. NH4+–N and NO3−–N concentrations were determined using a flow injection analyser, TP, TK, AK, and AP were measured using inductively coupled plasma mass spectrometry (ICP-MS) analysis (IRIS Intrepid II XSP system; Thermo Electric Co., USA). Soil pH was determined from a soil water (1:5 w/v) suspension, prepared by shaking 30 min, using a conductivity meter.

Total DOCpan> contenpan>t was measured using dichromate oxidationpan> titrationpan>[71], anpan>d EOC conpan>tenpan>t was anpan>alysed using 0.333 mol L−1 pan> class="Chemical">KMnO4 oxidation[72]. MBC content was measured using chloroform fumigation extraction[73].

Soil enzyme activity analysis

Soil amylase activity was measured using 2 g of fresh soil ipan class="Chemical">ncubated for 24 h at 37 °C according to Ebregt’s method[74]. Soil invertase activity was measured as at 30 °C and pH 4.65 in Na-acetate buffer according to Gianpan>freda’s method[75]. Soil cellulase activities were detected by anpan> incubationpan> accordinpan>g to Sharma’s method[76], and soil catalase activity was determined at pH 7.0, following the monpan>itoring of the decompositionpan> of pan> class="Chemical">H2O2 at 240 nm with an extinction coefficient of 43.6 M−1cm−1 according to Roggenkamp and Sahm[77].

Data analyses

All data were analysed usipan class="Chemical">ng SPSS 22.0 for Windows (SPSS Inc., Chicago, IL, USA). One-way analyses of variance (ANOVA) and comparisons among means were made using the least significant difference (LSD) test, with p < 0.05 regarded as significant. Pearson’s correlation coefficients of soil LOC fractions (EOC, DOC, anpan>d pan> class="Disease">MBC) with enzyme activities (invertase, cellulase, catalase, and amylase) and other soil characteristics were estimated, with a significance level of p < 0.05.