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REN Jiaxin, LIU Yang, WU Qifeng, et al. Effect of lime combined with biochar application on soil microbial activity in a Phyllostachys violascens plantation[J]. Journal of Zhejiang A&F University, 2026, 43(X): 1−10 doi:  10.11833/j.issn.2095-0756.20260254
Citation: REN Jiaxin, LIU Yang, WU Qifeng, et al. Effect of lime combined with biochar application on soil microbial activity in a Phyllostachys violascens plantation[J]. Journal of Zhejiang A&F University, 2026, 43(X): 1−10 doi:  10.11833/j.issn.2095-0756.20260254
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Effect of lime combined with biochar application on soil microbial activity in a Phyllostachys violascens plantation

DOI: 10.11833/j.issn.2095-0756.20260254
  • 1.

    College of Environmental and Resource Sciences/College of Carbon Neutrality, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China

  • 2.

    Bureau of Agriculture and Rural Affairs of Qujiang District, Quzhou 324022, Zhejiang, China

  • 3.

    Agroforestry Technology Extension Centre of Lin’an District, Hangzhou 311300, Zhejiang, China

  • Received Date: 2026-04-30
  • Accepted Date: 2026-07-03
  • Rev Recd Date: 2026-06-30
  •   Objective  This study aimed to investigate the effects of combined application of lime and biochar on soil enzyme activities and microbial carbon metabolism functions in a Lei bamboo (Phyllostachys violascens) plantation, so as to provide a reference for ameliorating soil acidification in these plantations.   Method  A field experiment was conducted in a Lei bamboo plantation with treatments of sole biochar application (10 and 20 t·hm−2) and their combination with lime (4 t·hm−2), using no amendment as the control. The effects of combined application of lime and biochar on soil chemical properties, enzyme activities, and microbial carbon source metabolism functions were analyzed.   Result  Compared with the control, sole lime application significantly increased soil pH, electrical conductivity, microbial biomass carbon and nitrogen contents, and the activities of β-glucosidase and leucine aminopeptidase (P＜0.05). Sole biochar application increased soil pH to a lesser extent than lime, but significantly enhanced soil organic carbon content and β-glucosidase activity (P＜0.05). All combined applications of biochar and lime significantly increased soil pH, organic carbon and microbial biomass carbon and nitrogen contents (P＜0.05), and improved the utilization rates of citric acid and aspartic acid as well as soil basal respiration rate. Sole biochar application significantly increased the bacterial ACE index and Shannon index (P＜0.05), while the lime treatment altered the bacterial community structure. Sole biochar, sole lime, and their combined application all significantly increased the fungal ACE index and Shannon index (P＜0.05). Partial least squares path modeling revealed that lime application increased soil basal respiration mainly by raising soil pH and microbial carbon source utilization rate, whereas biochar had a minor effect on soil basal respiration.   Conclusion  Combined application of lime and biochar can effectively reduce soil acidity in Lei bamboo plantation, synergistically increase soil organic carbon and microbial biomass, and enhance the microbial utilization capacity of carboxylic acid carbon sources. [Ch, 5 fig. 3 tab. 30 ref.]
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Effect of lime combined with biochar application on soil microbial activity in a Phyllostachys violascens plantation

doi: 10.11833/j.issn.2095-0756.20260254
  • 1. College of Environmental and Resource Sciences/College of Carbon Neutrality, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China
  • 2. Bureau of Agriculture and Rural Affairs of Qujiang District, Quzhou 324022, Zhejiang, China
  • 3. Agroforestry Technology Extension Centre of Lin’an District, Hangzhou 311300, Zhejiang, China
  • Received Date: 2026-04-30
  • Accepted Date: 2026-07-03
  • Rev Recd Date: 2026-06-30

Abstract:   Objective  This study aimed to investigate the effects of combined application of lime and biochar on soil enzyme activities and microbial carbon metabolism functions in a Lei bamboo (Phyllostachys violascens) plantation, so as to provide a reference for ameliorating soil acidification in these plantations.   Method  A field experiment was conducted in a Lei bamboo plantation with treatments of sole biochar application (10 and 20 t·hm−2) and their combination with lime (4 t·hm−2), using no amendment as the control. The effects of combined application of lime and biochar on soil chemical properties, enzyme activities, and microbial carbon source metabolism functions were analyzed.   Result  Compared with the control, sole lime application significantly increased soil pH, electrical conductivity, microbial biomass carbon and nitrogen contents, and the activities of β-glucosidase and leucine aminopeptidase (P＜0.05). Sole biochar application increased soil pH to a lesser extent than lime, but significantly enhanced soil organic carbon content and β-glucosidase activity (P＜0.05). All combined applications of biochar and lime significantly increased soil pH, organic carbon and microbial biomass carbon and nitrogen contents (P＜0.05), and improved the utilization rates of citric acid and aspartic acid as well as soil basal respiration rate. Sole biochar application significantly increased the bacterial ACE index and Shannon index (P＜0.05), while the lime treatment altered the bacterial community structure. Sole biochar, sole lime, and their combined application all significantly increased the fungal ACE index and Shannon index (P＜0.05). Partial least squares path modeling revealed that lime application increased soil basal respiration mainly by raising soil pH and microbial carbon source utilization rate, whereas biochar had a minor effect on soil basal respiration.   Conclusion  Combined application of lime and biochar can effectively reduce soil acidity in Lei bamboo plantation, synergistically increase soil organic carbon and microbial biomass, and enhance the microbial utilization capacity of carboxylic acid carbon sources. [Ch, 5 fig. 3 tab. 30 ref.]

REN Jiaxin, LIU Yang, WU Qifeng, et al. Effect of lime combined with biochar application on soil microbial activity in a Phyllostachys violascens plantation[J]. Journal of Zhejiang A&F University, 2026, 43(X): 1−10 doi:  10.11833/j.issn.2095-0756.20260254
Citation: REN Jiaxin, LIU Yang, WU Qifeng, et al. Effect of lime combined with biochar application on soil microbial activity in a Phyllostachys violascens plantation[J]. Journal of Zhejiang A&F University, 2026, 43(X): 1−10 doi:  10.11833/j.issn.2095-0756.20260254
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  • 土壤酸化是制约农业绿色发展的重要障碍因子之一。由于受长期不合理施肥和酸雨等影响，近30 a来中国土壤pH平均下降了0.5，且在南方地区更为突出[1]。据估算，酸化农田土壤面积约占全球耕地面积的50%，随着集约经营强度的增加，这一比例仍在不断上升[2]。土壤酸化不仅影响作物根系生长，而且抑制土壤微生物多样性和代谢功能，致使土壤养分供应功能紊乱，作物产量下降。因此，寻求高效的土壤酸化阻控方法对提升农产品产量与品质，促进农林生态系统可持续发展具有重要的实践意义。

    土壤微生物是土壤有机质分解和养分循环的重要驱动力，微生物的群落组成和活性可以一定程度反映土壤生态系统功能变化[3]。土壤β-葡萄糖苷酶、亮氨酸氨基肽酶和酸性磷酸酶等酶活性可表征土壤微生物群落对碳、氮、磷代谢需求，与土壤pH、养分可利用性和微生物群落结构的变化密切相关[4]。土壤微生物基础呼吸及其对葡萄糖、柠檬酸等碳源底物的利用速率可进一步表征土壤微生物代谢功能和有机质分解等过程。石灰和生物质炭是改良酸性土壤的重要材料，不仅可以中和土壤酸度，而且改善土壤结构和氮磷有效性[5−6]。曾廷廷等[7]通过整合分析发现：酸化土壤中施用石灰对粮食类作物、经济类作物增产率达42%和47%，且石灰输入量以3~6 t·hm−2为宜。生物质炭被认为是土壤酸性改良和增进土壤固碳减排的潜在途径，其较高的阳离子交换量和灰分含量使其有效中和土壤酸度。如不同原料生物质炭均能提高酸化土壤pH、有机碳含量以及酶活性，并能不同程度降低土壤交换性氢、交换性铝和交换性酸总量[5]。谭春玲等[8]研究发现：生物质炭的多孔结构可为微生物提供栖息地，其活性官能团可介导电子传递促进微生物代谢活性。AHMAD等[9]通过在酸性甘蔗Saccharum officinarum田中联合施用生物质炭与石灰显著提高了土壤pH、有机碳和酸性磷酸酶活性。

    雷竹Phyllostachys violascens是中国南方的一种优良笋用竹种。雷竹林覆盖和重施肥栽培技术具有较高的经济效益，但长期集约经营使雷竹林土壤酸化、养分失衡，导致土壤退化[10]。石灰输入是常见的酸化土壤改良技术。然而，长期施用石灰可能会加速土壤有机质以及表层腐殖质的矿化，从而引起土壤碳库损失[6]。生物质炭具有巨大的比表面积，可以有效保水保肥，吸附土壤中可利用氮，促进养分固持[11]。生物质炭输入土壤可有效提高土壤中细菌多样性，改变微生物群落结构，并通过提高土壤微生物量以及活性来改善土壤质量[12]。然而，生物质炭输入土壤的改良效果也因其材料、输入量及与不同物料的配施存在差异。包骏瑶等[10]研究发现：山核桃Carya cathayensis蒲壳炭和玉米Zea mays秸秆炭较竹炭对雷竹林酸化土壤具有较好的改良能力。然而，生物质炭与石灰配施如何影响土壤微生物代谢活性和群落组成尚不清楚，限制了生物质炭在竹林生态系统的推广应用。为此，本研究利用田间试验，设置生物质炭单施及生物质炭与石灰配施处理，探究不同用量生物质炭与石灰施用对雷竹林土壤酶活性、微生物碳源代谢功能及群落组成的影响，为雷竹林土壤酸化改良和生产力提升提供参考依据。

    • 1.   研究地区与研究方法

      1.1.   研究区概况

    • 研究区位于浙江省杭州市临安区太湖源镇(30°23′N，119°72′E)。该区属亚热带季风气候，丘陵地区，年平均气温为15.9 ℃，最高气温为41.3 ℃，最低气温为−13.3 ℃，年降水量约1 550.0 mm，无霜期为236.0 d。土壤类型属于粉砂岩母质上发育的红壤土类。试验区为长期集约经营雷竹林示范区，与相邻区域未集约经营林地相比，试验样地土壤pH下降了1.0~1.5个单位。试验地土壤基础化学性质：pH为4.40，有机碳为34.50 g·kg−1，全氮为3.30 g·kg−1，有效磷为315.00 mg·kg−1，速效钾为545.01 mg·kg−1。

      • 1.2.   试验设计与样品采集

    • 采用随机区组设计，样地共布置6个处理：①对照(不施生物质炭及石灰，ck)；②输入10 t·hm−2生物质炭处理(B1)；③输入20 t·hm−2生物质炭处理(B2)；④输入4 t·hm−2石灰(L)；⑤10 t·hm−2生物质炭和4 t·hm−2石灰共同施用(B1L)；⑥20 t·hm−2生物质炭和4 t·hm−2石灰共同施用(B2L)。每个处理3个重复，共包含18个小区，每个试验小区面积为40 m2 (10 m×4 m)。生物质炭为竹材边角料在600 ℃裂解2 h制成，过2 mm筛子备用。生物质炭的基本性质：pH为9.11，总碳为59.93%，全氮为0.86%，氢为2.42%，硫为0.27%(百分比为质量分数)，碳氮比69.69。

      田间试验半年后采集土壤样品，每个小区按照五点采样法采集并混合为一个样品。每份混合样品过2 mm筛子，挑去肉眼可见石粒、根系以及植物残体，并分成3份：一份室温自然风干，用于基本理化性质测定；一份鲜土直接存于4 ℃冰箱，用于微生物生物量、土壤酶活性和微生物碳源利用效率测定分析；另一份经冷冻干燥后于−70 ℃冰箱保存。

      • 1.3.   土壤理化性质和酶活性测定

    • 土壤pH、电导率(EC)、有机碳(SOC)、全氮(TN)测定参照鲁如坤[13]的研究。有机碳用重铬酸钾-硫酸外加热法进行测定；全氮用凯氏定氮法测定；土壤微生物生物量碳氮(MBC、MBN)采用氯仿熏蒸-K2SO4浸提法进行测定[13]。β-葡萄糖苷酶、亮氨酸氨基肽酶和酸性磷酸酶活性采用微孔板荧光法，其原理为利用底物与酶水解释放4-甲基伞形酮酰(4-MUB)进行荧光检测，通过荧光强度的变化反映酶活性[14]。用多功能酶标仪(Synergy™H1,Biotek)在荧光激发光365 nm和检测光波长450 nm下测定反应液荧光值。酶活性单位为nmol·g−1·h−1。

      • 1.4.   土壤微生物碳源利用速率测定

    • 微生物碳源利用速率(反映为基础呼吸和底物诱导呼吸)利用MicroResp方法测定[15−16]。该方法用原土进行培养，测定的底物诱导呼吸可以反映土壤的微生物碳源利用速率。该方法克服了依赖土壤悬浮液提取物和细胞后续生长状况条件制约，操作简便，反应灵敏。本研究选用葡萄糖、柠檬酸、天冬氨酸和丁香酸作为典型的碳源底物。碳源的最终添加质量浓度均为30 mg·mL−1。基础呼吸以无菌水代替底物测定。土壤碳源利用速率单位为μg·g−1·h−1，数值均以二氧化碳态碳释放量(CO2-C)计。

      • 1.5.   土壤DNA提取和高通量测序分析

    • 使用Powersoil DNA分离试剂盒，根据说明书进行DNA提取。细菌16S rRNA基因扩增采用515F和907R引物，真菌ITS基因扩增利用引物1737F和2043R。PCR反应后所得产物用质量浓度为2%的琼脂糖凝胶提取，并使用AxyPrep DNA凝胶提取试剂盒进一步纯化，使用QuantiFluorTM-ST进行定量检测，测序使用Illumina MiSeq平台完成。测序得到的具有条形码匹配的原始序列使用QIIME软件进行分析。用UPARSE软件对操作分类单元(OTU)进行聚类，相似性水平为97%。从每个OTU中选出最丰富的序列作为该OTU的代表性序列。根据UNITE数据库，使用RDP层级分类器对这些代表真菌ITS基因序列从属到门进行分类，置信阈值为70%。根据最小序列样本数原则，对群落结构进行多样性分析。

      • 1.6.   数据处理与统计分析

    • 采用SPSS22.0进行双因素方差分析(two-way ANOVA)，检验石灰和生物质炭施用间的差异及互作效应；采用单因素方差分析(one-way ANOVA) Duncan法多重比较检验不同处理间差异显著性，显著性水平为0.05。采用皮尔逊(Pearson)相关分析法进行相关性检验，显著性水平为0.05。采用偏最小二乘路径模型(PLS-PM)解析生物质炭与石灰施用对土壤微生物活性的调控路径。

    2.   结果与分析

      2.1.   土壤理化性质变化

    • 与对照相比，10和20 t·hm−2生物质炭单施处理，及其与石灰配施处理，均能显著提高有机碳及土壤微生物生物量碳质量分数(P＜0.05)；单施石灰可大幅提高土壤 pH、电导率、土壤微生物生物量碳氮。生物质炭与石灰配施处理的土壤pH与电导率均维持在较高水平，其中10 t·hm−2生物质炭与石灰配施处理显著提高了土壤微生物生物量碳氮比(P＜0.05)。双因素方差分析表明：生物质炭对土壤pH、有机碳、碳氮比、土壤微生物生物量碳及土壤微生物生物量碳氮比均存在显著(P＜0.05)或极显著影响(P＜0.001)；石灰处理则极显著提升了土壤pH、电导率、土壤微生物生物量碳氮(P＜0.001)，两者对土壤pH、有机碳、土壤微生物生物量碳氮存在显著交互效应(P＜0.05)。

      处理 pH 电导率/
      (mS cm−1)      		 有机碳/
      (g·kg−1)      		 全氮/
      (g·kg−1)      		 碳氮比 微生物生物量碳
      (MBC)/(mg·kg−1)      		 微生物生物量氮
      (MBN)/(mg·kg−1)      		 MBC/MBN
      ck 4.09±0.07 d 0.12±0.02 b 37.95±1.76 c 3.64±0.31 a 10.51±1.42 b 300.00±39.87 d 31.54±8.35 c 9.73±1.32 bc
      B1 4.28±0.06 cd 0.11±0.02 b 44.43±1.51 b 3.39±0.32 a 13.18±1.26 ab 744.28±111.59 bc 64.53±5.17 b 11.67±2.7 ab
      B2 4.48±0.20 c 0.11±0.03 b 49.03±4.34 a 3.43±0.25 a 14.43±2.34 a 642.52±99.17 c 67.16±10.02 b 9.83±2.76 bc
      L 7.45±0.03 a 0.20±0.06 a 36.77±1.88 c 3.45±0.11 a 10.68±0.78 b 954.37±179.53 a 116.58±21.81 a 8.19±0.54 b
      B1L 7.00±0.15 b 0.18±0.05ab 46.96±0.91 ab 3.67±0.46 a 12.94±1.80 ab 903.54±39.26 ab 63.19±3.16 b 14.32±0.83 a
      B2L 7.32±0.12 a 0.22±0.03 a 44.12±0.32 b 3.28±0.31 a 13.51±1.25 ab 622.66±59.55 c 56.93±6.09 b 10.96±0.59 bc
      B * ns *** ns *** * ns *
      L *** *** ns ns ns *** *** ns
      B×L *** ns * ns ns *** *** ns
      　　说明：ck. 对照；B1. 10 t·hm−2生物质炭；B2. 20 t·hm−2生物质炭；L. 石灰。B×L. 石灰与生物质炭的互作效应。同列不同小写字母表示差异显著(P＜0.05)。*表示P＜0.05；***表示P＜0.001；ns表示不显著。

      Table 1.  Effect of biochar and lime application on soil physicochemical properties and two-way ANOVA

      • 2.2.   土壤酶活性变化

    • 与对照相比，生物质炭单施能显著提升β-葡萄糖苷酶和酸性磷酸酶活性(P＜0.05)，其中20 t·hm−2生物质炭处理显著提高了亮氨酸氨基肽酶活性(P＜0.05)。单施石灰显著提升β-葡萄糖苷酶与亮氨酸氨基肽酶活性(P＜0.05)，却显著降低了酸性磷酸酶活性(P＜0.05)。10 t·hm−2生物质炭与石灰配施处理维持了较高的β-葡萄糖苷酶和亮氨酸氨基肽酶活性；20 t·hm−2生物质炭与石灰配施则降低了β-葡萄糖苷酶活性。双因素方差分析表明：生物质炭和石灰对β-葡萄糖苷酶、亮氨酸氨基肽酶及酸性磷酸酶活性均存在显著影响(P＜0.05)；两者对β-葡萄糖苷酶和亮氨酸氨基肽酶活性表现为极显著交互效应(P＜0.001)。

      处理 β-葡萄糖苷酶活性/
      (nmol·g−1·h−1)      		 亮氨酸氨基肽酶
      活性/(nmol·g−1·h−1)      		 酸性磷酸酶活性/
      (nmol·g−1·h−1)
      ck 25.34±1.26 d 11.54±0.65 c 168.70±14.30 bc
      B1 34.68±0.74 a 11.86±0.27 c 197.87±21.37 a
      B2 31.08±0.41 bc 16.27±0.70 b 181.53±9.64 ab
      L 29.20±1.47 c 17.67±0.30 a 95.38±6.54 d
      B1L 32.71±3.22 ab 18.6±0.66 a 154.25±12.89 c
      B2L 22.92±1.64 d 15.64±0.37 b 95.44±9.26 d
      B *** *** *
      L * *** *
      B×L *** *** ns
      　　说明：ck. 对照；B1. 10 t·hm−2生物质炭；B2. 20 t·hm−2生物质炭；L. 石灰。B×L. 石灰与生物质炭的互作效应。同列不同小写字母表示差异显著(P＜0.05)。*表示P＜0.05；***表示P＜0.001；ns表示不显著。

      Table 2.  Effect of biochar and lime application on soil enzyme activities and two-way ANOVA

      • 2.3.   土壤微生物碳源代谢功能的变化

    • 与对照相比，单施生物质炭未改变土壤基础呼吸，却显著降低了葡萄糖与丁香酸的利用速率(P＜0.05)；单施石灰显著提升了土壤基础呼吸、柠檬酸和天冬氨酸的利用速率(P＜0.05)。生物质炭与石灰配施处理的土壤基础呼吸与单施石灰相似；但显著降低了葡萄糖利用速率(P＜0.05)。石灰与生物质炭配施使柠檬酸、天冬氨酸利用速率显著高于对照及生物质炭处理(P＜0.05)。双因素方差分析显示：生物质炭和石灰对土壤基础呼吸、葡萄糖及天冬氨酸利用速率有显著互作效应(P＜0.05)。

      处理
      		 土壤基础呼吸/
      (μg·g−1·h−1)      		 葡萄糖利用速率/
      (μg·g−1·h−1)      		 柠檬酸利用速率/
      (μg·g−1·h−1)      		 天冬氨酸利用速率/
      (μg·g−1·h−1)      		 丁香酸利用速率/
      (μg·g−1·h−1)
      ck 0.57±0.15 b 2.13±0.26 a 1.00±0.31 c 0.31±0.16 c 1.26±0.29 ab
      B1 0.32±0.11 b 1.45±0.22 b 0.33±0.12 c 0.32±0.08 c 0.28±0.03 c
      B2 0.24±0.09 b 0.44±0.07 c 0.50±0.15 c 0.50±0.05 c 0.37±0.15 c
      L 1.23±0.26 a 1.94±0.12 a 5.81±0.53 a 5.01±0.21 a 1.43±0.40 a
      B1L 1.19±0.24 a 0.48±0.13 c 3.79±0.71 b 3.10±0.40 b 0.50±0.03 c
      B2L 1.18±0.19 a 0.32±0.09 c 4.28±0.65 b 3.26±0.28 b 0.94±0.26 b
      B * *** * *** ***
      L *** *** *** *** *
      B×L * * ns *** ns
      　　说明：ck. 对照；B1. 10 t·hm−2生物质炭；B2. 20 t·hm−2生物质炭；L. 石灰。B×L. 石灰与生物质炭的互作效应。同列不同小写字母表示差异显著(P＜0.05)。*表示P＜0.05；***表示P＜0.001；ns表示不显著。

      Table 3.  Effect of biochar and lime application on the functional diversity of microbial carbon metabolism in soil and two-way ANOVA

      • 2.4.   土壤细菌和真菌群落组成的变化

    • 与对照相比，单施生物质炭降低了放线菌门Actinobacteria相对丰度，提高了粘细菌门Myxococcota相对丰度，而单施石灰及其与生物质炭配施处理均降低了放线菌门和酸杆菌门Acidobacteria相对丰度，但提高了绿弯菌门Chloroflexi相对丰度(图1A)。对真菌群落，施用20 t·hm−2生物质炭处理降低了子囊菌门Ascomycota相对丰度，提高了担子菌门Basidiomycota相对丰度；单施石灰提高了子囊菌门相对丰度，石灰与生物质炭配施提高了担子菌门相对丰度(图1B)。

      Figure 1.  Effect of co-application of lime and biochar on the relative abundance of soil bacterial (A) and fungal (B) communities

      主坐标分析(PCoA)表明：石灰处理对土壤细菌群落结构具有显著影响(P＜0.001)，群落结构根据石灰处理与否在第1轴上存在明显的分异(图2A)。相似性分析(ANOSIM)表明：生物质炭与石灰处理对细菌群落有显著影响(图2A，P＜0.001)，而生物质炭与石灰对真菌群落影响较细菌小(图2B)。

      Figure 2.  Principal Coordinate Analysis (PCoA) of the effect of co-application of lime and biochar on the structure of soil bacterial (A) and fungal (B) communities

      如图3所示：与对照相比，生物质炭与石灰处理均显著提高了细菌的ACE指数(P＜0.05)，而10和20 t·hm−2生物质炭处理显著提高了细菌Shannon指数(P＜0.05)。与对照相比，单施生物质炭、石灰及其两者配施处理均能显著提高真菌的ACE指数和Shannon指数(P＜0.05)。

      Figure 3.  Analysis of diversity indices for soil bacterial (A, C) and fungal (B, D) communities under co-application of lime and biochar

      • 2.5.   影响土壤基础呼吸与酶活性的主要驱动因子与路径

    • Mantel分析(图4)表明：亮氨酸氨基肽酶与真菌ACE指数呈显著正相关(P＜0.05)，酸性磷酸酶与土壤pH、电导率、柠檬酸利用速率、天冬氨酸利用速率呈显著正相关(P＜0.001)，土壤基础呼吸速率与电导率、柠檬酸利用速率、细菌Shannon指数呈显著正相关(P＜0.05)。

      Figure 4.  Mantel analysis between soil basal respiration and enzyme activities

      采用偏最小二乘路径模型(PLS-PM)解析生物质炭与石灰施用对土壤微生物活性的调控路径(图5)。结果表明：石灰施用对土壤pH具有极显著正向直接效应(P＜0.001)，土壤pH对碳源利用速率(P＜0.05)和微生物多样性(P＜0.001)表现为显著正向调控，碳源利用速率对土壤基础呼吸具有显著正向直接效应(P＜0.05)，是驱动土壤呼吸增强的直接因素；而生物质炭、土壤养分及微生物多样性对土壤呼吸的效应均不显著。

      Figure 5.  Partial least squares path modeling (PLS-PM) of the drivers of soil basal respiration

    3.   讨论

      3.1.   石灰和生物质炭施用对土壤化学性质和微生物量的影响

    • 本研究表明：施用石灰可提高土壤pH，而施用生物质炭对pH提高幅度较低。这与石灰呈强碱性，可以有效中和土壤潜性酸和活性酸，从而迅速降低土壤酸度有关[17]。生物质炭对土壤pH提高程度较低，这与生物质炭的材料性质、施用量与裂解温度有关，如竹炭相比秸秆炭具有更低的酸中和能力[18]。然而，生物质炭与石灰相比，可显著提高土壤有机碳含量。这与生物质炭具有高度芳香性的碳组分和碳氮比有关。相反，单施石灰并未导致土壤有机碳含量下降。有研究表明：石灰施入也可能通过提高土壤pH，缓解酸胁迫对微生物的抑制作用，促进有机碳矿化分解，加速土壤有机质损失[19−20]。生物质炭、石灰单施及配施均能显著增加土壤微生物生物量碳氮。研究表明：生物质炭疏松多孔结构可为微生物提供居住环境，且生物质炭自身含有少量氮磷元素，可为微生物生长提供养分[21]。芮绍云等[22]研究发现：单施生物质炭或过氧化钙均能显著提高红壤微生物生物量碳氮。微生物生物量碳氮能反映土壤碳氮有效性变化，又能反映微生物碳氮需求[23]。生物质炭与石灰联合输入极大程度提高土壤pH，可能导致养分有效性增强，从而可能促进了微生物的增殖。

      • 3.2.   石灰和生物质炭施用对土壤酶活性的影响

    • 土壤碳氮磷转化酶活性可反映土壤养分转化速率和微生物养分需求状况[24]。β-葡萄糖苷酶是与纤维素降解相关的关键酶，亮氨酸氨基肽酶与土壤中氮转化密切相关，酸性磷酸酶则与土壤中有机磷素转化有关[25]。本研究中，不同酶活性对生物质炭和石灰输入的响应不一致。包骏瑶等[10]研究表明：玉米秸秆炭、山核桃蒲壳炭、竹炭均能显著提高雷竹林土壤中β-葡萄糖苷酶和酸性磷酸酶活性，而对亮氨酸氨基肽酶活性无影响。包建平等[26]研究发现：生物质炭单施对南方红壤β-葡萄糖苷酶、酸性磷酸酶以及亮氨酸氨基肽酶活性均无显著影响。本研究中10 t·hm−2生物质炭处理显著提高了β-葡萄糖苷酶活性，而20 t·hm−2生物质炭处理对β-葡萄糖苷酶、亮氨酸氨基肽酶和酸性磷酸酶活性无显著影响，这表明生物质炭对酶活性的影响与生物质炭的施用量有关。然而，石灰单施及其与20 t·hm−2生物质炭配施均能显著降低β-葡萄糖苷酶活性，表明石灰输入是引起β-葡萄糖苷酶活性降低的主因，这可能与较高的土壤pH抑制酶活性有关。邱全敏等[27]研究发现：石灰施入酸化荔枝Litchi chinensis园土壤，可降低酸性磷酸酶的活性，这与本研究结果相似。因此，生物质炭与石灰配施对酶活性的影响与酶活性种类、施用量和土壤养分变化密切相关。

      • 3.3.   石灰和生物质炭施用对土壤微生物群落组成及碳源代谢功能的影响

    • 主坐标分析表明：生物质炭与石灰配施处理对细菌群落组成和结构有显著影响，而对真菌群落影响较细菌小，表明细菌对生物质炭和石灰添加的响应更加灵敏。相比生物质炭，石灰处理对细菌的影响要高于生物质炭处理。单施生物质炭显著降低了放线菌门相对丰度，提高了粘细菌门相对丰度，而单施石灰处理显著降低了放线菌门和酸杆菌门相对丰度，表明不同物种对生物质炭和石灰的响应差别较大。生物质炭单施显著提高了细菌的多样性指数，而生物质炭与石灰配施更有利于提高真菌的多样性。这可能是生物质炭与石灰配施可以更好地改良土壤酸性，提高外源碳输入，促进真菌类群的多样性[28]。微生物碳源代谢功能是土壤微生物介导碳输出的主要反映。CHEN等[29]研究表明：土壤呼吸速率随着生物质炭的增加而降低，这与本研究单施生物质炭降低土壤基础呼吸速率的结果一致，可能是因为生物质炭在土壤中存在一段时间后，与土壤有机质及矿物形成稳定复合体，保护有机质免被微生物和酶分解。但石灰施用使土壤基础呼吸速率和碳源底物利用速率均明显提高，说明石灰对于土壤微生物碳源代谢功能有较强的主导作用。淡俊豪等[30]研究表明：生石灰输入酸性土壤中能够显著提高土壤pH，改善土壤微环境，提高土壤微生物代谢活性和碳源利用能力。与此相似，本研究中雷竹林生物质炭单施未显著提高微生物碳源利用速率，但配施石灰显著提高了微生物对多种碳源的利用速率，增强了微生物代谢活性。这表明石灰相比生物质炭可以更有效促进微生物对羧酸、氨基酸、酚酸类等碳源的利用。偏最小二乘路径模型进一步证实：生物质炭与石灰施用对土壤微生物代谢功能影响效应存在明显差异。石灰施用显著提升了土壤pH，加快了微生物碳源的利用速率，从而提高了土壤基础呼吸，而生物质炭对微生物多样性和碳源代谢无显著影响，这也印证了这一解释。

    4.   结论

    • 单施石灰显著提高了土壤pH和微生物生物量，并显著改变了细菌群落结构，但对有机碳含量无影响。单施生物质炭显著提高雷竹林土壤有机碳含量和碳转化酶活性，但降低了微生物对葡萄糖和丁香酸的利用速率。石灰与生物质炭配施能有效降低雷竹林土壤酸度，同时提高了土壤有机碳和微生物生物量，并提高了微生物对羧酸类碳源利用能力。石灰配施引起的土壤pH升高，是驱动土壤生物碳源代谢活性的关键因子。因此，生物质炭和石灰配施可有效缓解雷竹林土壤酸化，提升土壤有机碳含量和微生物活性。

Reference (30)

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