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林业科学 ›› 2019, Vol. 55 ›› Issue (8): 106-117.doi: 10.11707/j.1001-7488.20190812

• 论文与研究报告 • 上一篇    下一篇

凉水国家自然保护区不同林型红松林土壤nosZ型反硝化微生物群落组成和多样性分析

陈秀波1,2, 朱德全3,4, 赵晨晨3, 张路路3,4, 陈立新1, 段文标1   

  1. 1. 东北林业大学林学院 哈尔滨 150040;
    2. 贵州财经大学艺术学院 贵阳 550025;
    3. 佳木斯大学理学院 佳木斯 154007;
    4. 佳木斯大学中-乌农林技术开发与应用国际联合实验室 佳木斯 154007
  • 收稿日期:2018-12-24 修回日期:2019-05-30 发布日期:2019-09-05
  • 基金资助:
    国家自然科学基金项目“阔叶红松林连根拔起倒木及其丘坑微立地特征及对幼苗更新的影响”(31670627)和“阔叶红松林凋落物-氮磷沉降耦合输入对土壤团聚体碳氮激发效应及稳定性影响”(31770656);黑龙江省省属本科高校基本科研业务费项目“驱动东北典型红松混交林土壤碳氮循环的关键微生物群落结构和分子生态学特征”(2017-KYYWF-0597)。

Structure and Diversity of Soil nosZ-Type Denitrifying Microbial Community in Different Types of Pinus koraiensis Forests in Liangshui National Nature Reserve

Chen Xiubo1,2, Zhu Dequan3,4, Zhao Chenchen3, Zhang Lulu3,4, Chen Lixin1, Duan Wenbiao1   

  1. 1. College of Forestry, Northeast Forestry University Harbin 150040;
    2. College of Art, Guizhou University of Finance and Economics Guiyang 550025;
    3. College of Science, Jiamusi University Jiamusi 154007;
    4. China-Ukraine International Joint Laboratory for Agricultural and Forestry Technology Development and Application, Jiamusi University Jiamusi 154007
  • Received:2018-12-24 Revised:2019-05-30 Published:2019-09-05

摘要: [目的]分析凉水国家级自然保护区内的3种原始红松林(云冷杉红松林、椴树红松林和枫桦红松林)、红松人工林和红松天然次生林5种林型的土壤nosZ型反硝化微生物的群落组成和多样性特征,为全面了解不同林型红松林土壤的反硝化潜势和氮循环过程提供数据支持。[方法]以选取的5种林型红松林林下土壤为研究对象,以反硝化过程中的关键酶——氧化亚氮还原酶的编码基因nosZ为标记基因,采用高通量测序和生物信息学分析技术进行研究。[结果]从5种林型红松林15个土壤样品中一共得到nosZ基因631 878条有效序列,579 871条优质序列,长度分布在178~383 bp之间,主要分布在260 bp。5种林型红松林土壤nosZ型反硝化微生物主要门类为变形菌门和拟杆菌门,核心属为伯霍尔德杆菌属、黄杆菌属、慢生根瘤菌属、假单胞菌属、Dechloromonas属、芽单胞菌属、无色杆菌属和中华根瘤菌属。nosZ型反硝化微生物α多样性分析显示:除枫桦红松林的Shannon和Simpson指数显著高于红松天然次生林外,5种林型红松林之间土壤nosZ型反硝化菌群的4种α多样性指数(Shannon、Chao1、ACE和Simpson指数)差异不显著。β多样性分析显示:5种林型土壤nosZ型反硝化微生物群落组成差异显著(R=0.387,P=0.006),但3种原始红松林之间差异不显著。土壤铵氮和全氮含量是显著影响nosZ型反硝化微生物群落的主要因子(P<0.05)。[结论] 5种林型红松林土壤nosZ型反硝化微生物多数α多样性指数无显著差异,但β多样性差异显著,引起不同林型之间nosZ型反硝化微生物组成和丰度的主要环境因子是土壤铵氮和全氮含量。

关键词: 原始红松林, 人工林, 天然次生林, nosZ基因, 高通量测序, 反硝化微生物

Abstract: [Objective] The community structure and diversity characteristics of soil nosZ-type denitrification microorganisms in the three kinds of primitive Pinus koraiensis forest (Picea koraiensis-Abies nephrolepis-Pinus koraiensis forest, Tilia spp. and Pinus koraiensis forest and Betula costata-Pinus koraiensis forest), Pinus koraiensis plantation and natural Pinus koraiensis secondary forest in Liangshui national nature reserve were analyzed to provide data support for a comprehensive understanding of the denitrification potential of different types of Pinus koraiensis forests and the nitrogen cycle process in the soil.[Method] The understory soils of five types of Pinus koraiensis forests were selected as the research object. The nosZ gene encoding nitrous oxide reductase (a key enzyme in the process of denitrification) of denitrification microorganisms was used as marker gene. The community composition and diversity characteristics of nosZ-type denitrification microorganisms in the soil were analyzed by high-throughput sequencing and bioinformatics analysis tequniques.[Result]The results showed that a total of 631 878 effective sequences of nosZ gene were obtained by high throughput sequencing from 15 soil samples in the five types of Pinus koraiensis forests, among which 579 871 sequences were high quality. These sequences were distributed between 178-383 bp but were mainly distributed around 260 bp. The main phyla of soil nosZ type denitrification microorganisms in five types of Pinus koraiensis forests were proteobacteria and bacteroidetes at the phylum level. However, the core genus included Burkholderia, Flavobacterium, Bradyrhizobium, Pseudomonas, Dechloromonas, Gemmatimonas, Achromobacter and Sinorhizobium. The analysis of alpha diversity of nosZ denitrifying microorganisms showed that the four alpha diversity index (Shannon, Chao1, ACE and Simpson index) of soil nosZ-type denitrification microorganisms among the five forest types were not siginificantly different, except that the Shannon and Simpson index of Betula costata-Pinus koraiensis forest was significantly higher than that of Pinus koraiensis natural secondary forest. Beta diversity analysis showed that there were significant differences in the composition of nosZ microbial communities in soils of five forest types (R=0.537,P=0.001), but there was no significantly different in the composition of three primitive Pinus koraiensis forests. Soil ammonia nitrogen and total nitrogen content were the main factors that significantly affected the nosZ type of denitrification microbial community (P<0.05).[Conclusion]There was no significant difference in the most alpha diversity index of soil nosZ-type denitrification microorganism, but beta diversity analysis showed that there were significant differences in the composition of nosZ microbial communities in soils of five forest types. The main environmental factors which influenced the composition and abundance of nosZ denitrification microorganisms over different forest types were soil ammonium nitrogen and total nitrogen.

Key words: primitive forest of Pinus koraiensis, plantation, natural secondary forest, nosZ gene, high-throughput sequencing, nitrifying microorganism

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