林业科学 ›› 2020, Vol. 56 ›› Issue (8): 27-37.doi: 10.11707/j.1001-7488.20200804
王磊,梁艺凡,杨军钱,张冰冰,王涛,施秀珍,胡行伟,黄志群*
收稿日期:
2019-05-05
出版日期:
2020-08-25
发布日期:
2020-09-15
通讯作者:
黄志群
基金资助:
Lei Wang,Yifan Liang,Junqian Yang,Bingbing Zhang,Tao Wang,Xiuzhen Shi,Hangwei Hu,Zhiqun Huang*
Received:
2019-05-05
Online:
2020-08-25
Published:
2020-09-15
Contact:
Zhiqun Huang
摘要:
目的: 探讨我国亚热带人工林主要造林树种对土壤氮循环相关功能微生物基因丰度和群落结构的影响,阐明土壤氮保留影响的微生物驱动机制的树种差异。方法: 对生长在酸性红壤上的5种人工林采用实时荧光定量PCR研究森林土壤的固氮微生物(nifH)、硝化微生物(AOA amoA、AOB amoA)、反硝化微生物(narG、nirK、nirS、nosZ)、真菌(ITS)和细菌(16S rRNA)基因丰度,通过建立克隆文库和测序,对AOA不同的末端限制性片段进行系统发育分析,通过采用末端限制性片段多态性技术(T-RFLP)分析AOA群落结构,研究各树种人工林土壤的保氮能力及相关功能微生物特征。结果: 1)5种人工林土壤pH值范围为4.63~4.82,木荷和火力楠林的土壤硝态氮含量显著高于闽楠和福建柏林(P < 0.05),火力楠和木荷林土壤的微生物生物量(MBC、MBN)在5个人工林中均最高,而闽楠和杉木林的均最低;凋落物的木质素和纤维素含量显著抑制土壤微生物生物量;2)5种森林土壤的固氮微生物nifH基因丰度无显著差异。木荷林的AOA丰度显著高于闽楠、火力楠和福建柏林,木荷林的AOB丰度显著高于福建柏林,木荷林的AOA和AOB丰度均最高;在氨氧化微生物中,AOA/AOB>2,AOA在数量上占据优势地位。反硝化微生物的narG、nirK和nosZ丰度在树种间均无显著差异,木荷林土壤的nirS丰度显著高于火力楠、杉木和福建柏林。闽楠林土壤的真菌ITS丰度显著高于木荷林,土壤细菌16S丰度在树种间均无显著差异;3)Pearson相关分析发现,土壤pH值与AOB、narG、nosZ显著正相关,土壤NO3--N含量与土壤AOA极显著正相关(P < 0.01);4)系统发育分析显示,5种森林土壤AOA类群属于Nitrosopumilus和Nitrososphaera,使用HpyCH4V限制性内切酶对AOA amoA基因PCR产物进行酶切,能够产生4个TRFs,其中TRF-76和TRF-165是最主要的2个片段类型,分别占总片段的54.88%~100%、0~45.12%,TRF-76和TRF-165的相对丰度的树种差异非常明显。通过非度量多维测度法(NMDS)分析,5种森林土壤AOA amoA的群落结构无显著差异(P>0.05)。结论: 亚热带酸性森林土壤中,AOA在数量上占据优势地位,在土壤硝化作用中可能发挥着主导作用;木荷林的AOA和AOB丰度均最高,nirS的基因丰度显著高于火力楠、杉木和福建柏林,木荷林的AOA丰度显著高于闽楠林,可能加剧氮素从森林生态系统损失的风险。5种森林土壤的AOA类群属于Nitrosopumilus和Nitrososphaera,AOA群落结构无显著差异。
中图分类号:
王磊,梁艺凡,杨军钱,张冰冰,王涛,施秀珍,胡行伟,黄志群. 亚热带主要造林树种土壤氮保留及相关功能的微生物特征[J]. 林业科学, 2020, 56(8): 27-37.
Lei Wang,Yifan Liang,Junqian Yang,Bingbing Zhang,Tao Wang,Xiuzhen Shi,Hangwei Hu,Zhiqun Huang. Characteristics of Soil nitrogen Retention and Related Functional Microorganism in Soils of Main Afforestation Species in Subtropical Region[J]. Scientia Silvae Sinicae, 2020, 56(8): 27-37.
表1
不同树种人工林基本概况(均值±标准误, n=3)①"
指标Items | 闽楠 | 火力楠 | 木荷 | 杉木 | 福建柏 |
P. bournei | M. macclurei | S. superba | C. lanceolata | F. hodginsii | |
林龄Stand age/a | 35 | 32 | 32 | 23 | 35 |
树高Tree height/m | 14.3±0.7 | 16.9±0.2 | 15.7±1.1 | 14.8±0.5 | 13.3±0.5 |
胸径DBH/cm | 14.7±0.4 | 18.8±1.1 | 23.1±5.4 | 19.8±2.4 | 21.6±2.7 |
林分密度Stand density/(hm-2) | 2 067 | 2 100 | 2 000 | 2 100 | 2 800 |
坡度Slope/(°) | 29 | 36 | 32 | 26 | 29 |
海拔Altitude/m | 425 | 412 | 512 | 330 | 547 |
凋落物碳含量Litter C/(g·kg-1) | 500.1±1.6bc | 492.1±1.6c | 505.5±2.3b | 495.6±2.4c | 514.5±1.4a |
凋落物氮含量Litter N/(g·kg-1) | 9.1±1.6ab | 7.6±1.4ab | 11.6±0.7a | 5.4±0.3b | 11.9±1.0a |
凋落物碳/氮Litter C/N | 58.1±9.0bc | 69.0±10.6b | 43.7±2.4c | 92.5±6.5a | 43.7±3.8c |
凋落物木质素Litter lignin/(g·kg-1) | 225.9±3.0ab | 145.1±19.7c | 157.8±9.9c | 261.0±17.9a | 220.2±4.4b |
凋落物纤维素含量Litter cellulose/(g·kg-1) | 314.4±5.3a | 274.3±4.3b | 209.4±11.5c | 310.7±22.0ab | 276.9±10.7ab |
细根碳含量Fine root C/(g·kg-1) | 441.6±14.9ab | 426.3±13.9b | 444.7±9.7ab | 354.7±12.1c | 472.3±14.1a |
细根氮含量Fine root N/(g·kg-1) | 10.8±0.2b | 16.9±1.1a | 7.4±0.8c | 8.2±0.2c | 7.8±0.6c |
细根碳/氮Fine root C:N | 41.0±0.9b | 25.4±0.9c | 61.7±8.3a | 43.5±1.8b | 61.5±5.4a |
表2
氮循环微生物定量PCR引物"
目的基因 Primer target | 引物名称 Primer name | 引物序列(5’-3’) Primer sequence (5’-3’) | 参考文献 Reference |
nifH | PloyF | TGCGAYCCSAARGCBGACTC | ( |
PloyR | ATSGCCATCATYTCRCCGGA | ||
AOA amoA | CrenamoA23f | ATGGTCTGGCTWAGACG | ( |
CrenamoA616 r | GCCATCCATCTGTATGTCCA | ||
AOB amoA | amoA1F | GGGGTTTCTACTGGTGGT | ( |
amoA2R | CCCCTCKGSAAAGCCTTCTTC | ||
nirK | nirK-F1aCu | ATCATGGTSCTGCCGCG | ( |
nirK-R3Cu | GCCTCGATCAGRTTGTGGTT | ||
nirS | Nirs-Cd3aF | GTSAACGTSAAGGARACSGG | ( |
Nirs-R3cd | GASTTCGGRTGSGTCTTGA | ||
narG | NarGG-F | TCGCCSATYCCG GCSATGTC | ( |
NarGG-R | GAGTTGTACCAGTCRGCSGAYTCSG | ||
nosZ | nosZ2F | CGCRACGGCAASAAGGTSMSSGT | ( |
nosZ2R | CAKRTGCAKSGCRTGGCAGAA | ||
ITS | ITS-1F | TCCGTAGGTGAACCTGCGG | ( |
5.8 s | CGCTGCGTTCTTCATCG | ( | |
16S rRNA | 519F | CAGCMGCCGCGGTANWC | ( |
907R | CCGTCAATTCMTTTRAGTT | ( |
表3
各树种人工林土壤基本理化性质(均值±标准误, n=3)"
土壤性质 Soil characteristics | 闽楠 | 火力楠 | 木荷 | 杉木 | 福建柏 |
P. bournei | M. macclurei | S. superba | C. lanceolata | F. hodginsii | |
黏粒Clay(%) | 20.31±2.99 | 15.89±3.64 | 16.06±1.64 | 15.72±3.14 | 19.71±0.69 |
粉粒Silt(%) | 48.07±1.90 | 41.74±5.37 | 32.76±2.41 | 47.30±5.17 | 37.97±6.16 |
砂粒Sand(%) | 31.62±2.07b | 42.38±7.62ab | 51.18±4.05a | 36.98±7.45b | 42.32±5.48ab |
密度Density/(g·cm-3) | 1.22±0.11 | 1.12±0.02 | 1.17±0.08 | 1.19±0.04 | 1.09±0.10 |
含水量Moisture content/(g·kg-1) | 253.20±9.68a | 231.64±10.41ab | 243.63±17.67a | 246.58±3.55a | 211.85±16.45b |
酸碱度pH | 4.75±0.02 | 4.69±0.11 | 4.82±0.11 | 4.69±0.21 | 4.63±0.03 |
全碳TC/(g·kg-1) | 24.82±4.05ab | 22.02±1.07ab | 22.56±5.16ab | 17.60±1.09b | 28.37±3.8a |
全氮TN/(g·kg-1) | 1.74±0.21 | 1.75±0.14 | 1.74±0.24 | 1.44±0.10 | 1.84±0.18 |
碳氮比C/N | 14.16±0.57 | 12.63±0.60 | 12.84±1.19 | 12.27±0.64 | 15.38±0.61 |
铵态氮NH4+-N/(mg·kg-1) | 10.66±0.68 | 10.92±2.56 | 10.11±1.08 | 10.04±0.99 | 11.77±2.85 |
硝态氮NO3--N/(mg·kg-1) | 0.37±0.09b | 4.24±1.04a | 4.64±2.21a | 2.08±0.69ab | 0.71±0.39b |
表3
各树种森林土壤理化性质及微生物生物量的Pearson相关分析①"
凋落物碳 Litter C | 凋落物氮 Litter N | 凋落物碳/氮 Litter C/N | 木质素 Lignin | 纤维素 Cellulose | 细根碳 Fine root C | 细根氮 Fine root N | 细根碳/氮 Fine root C/N | |
全碳TC | 0.43 | 0.35 | -0.44 | 0.01 | -0.19 | 0.69** | -0.06 | 0.26 |
全氮TN | 0.20 | 0.24 | -0.35 | -0.17 | -0.34 | 0.63* | 0.08 | 0.15 |
碳/氮C/N | 0.60* | 0.43 | -0.47 | 0.21 | 0.06 | 0.59* | -0.17 | 0.28 |
酸碱度pH | 0.01 | 0.26 | -0.33 | 0.04 | -0.46 | 0.25 | -0.14 | 0.22 |
铵态氮NH4+-N | 0.194 | 0.18 | -0.25 | 0.02 | -0.01 | 0.24 | 0.08 | 0.07 |
硝态氮NO3--N | -0.28 | -0.05 | 0.03 | -0.61* | -0.52* | -0.07 | 0.16 | 0.03 |
微生物生物量碳MBC | 0.05 | 0.26 | -0.31 | -0.80** | -0.67** | 0.50 | 0.23 | 0.15 |
微生物生物量氮MBN | 0.23 | 0.35 | -0.39 | -0.54* | -0.63* | 0.54* | 0.08 | 0.21 |
表4
各树种土壤氮循环功能微生物基因拷贝数与土壤理化性质的Pearson相关分析"
nifH | AOA amoA | AOB amoA | nirK | nirS | narG | nosZ | ITS | 16S | |
全碳TC | 0.10 | -0.54* | -0.13 | 0.28 | 0.15 | 0.20 | 0.26 | 0.20 | 0.20 |
全氮TN | 0.03 | -0.31 | 0.07 | 0.39 | 0.19 | 0.45 | 0.26 | 0.20 | 0.36 |
碳/氮C/N | 0.16 | -0.69** | -0.35 | 0.10 | 0.07 | -0.16 | 0.19 | 0.19 | -0.01 |
酸碱度pH | 0.30 | 0.13 | 0.57* | 0.50 | 0.49 | 0.64* | 0.64* | -0.14 | 0.35 |
铵态氮NH4+-N | -0.01 | -0.56* | -0.22 | -0.32 | -0.21 | -0.37 | -0.14 | 0.03 | -0.29 |
硝态氮NO3--N | -0.13 | 0.81** | 0.27 | -0.08 | -0.02 | 0.24 | -0.28 | -0.08 | 0.14 |
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