Community structure and diversity of soil nirK-type denitrifying microorganisms in three forest types of primitive Pinus koraiensis mixed forest in Liangshui National Nature Reserve, Lesser Khingan Mountains

doi:10.12302/j.issn.1000-2006.201902033

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2021, Vol. 45 ›› Issue (2): 77-86.doi: 10.12302/j.issn.1000-2006.201902033

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Community structure and diversity of soil nirK-type denitrifying microorganisms in three forest types of primitive Pinus koraiensis mixed forest in Liangshui National Nature Reserve, Lesser Khingan Mountains

CHEN Xiubo¹^,²(), DUAN Wenbiao¹^,^*(), CHEN Lixin¹, ZHU Dequan³^,⁴, ZHAO Chenchen³, LIU Dongxu³^,⁴

1. College of Forestry, Northeast Forestry University, Harbin 150040, China
2. College of Art, Guizhou University of Finance and Economics, Guiyang 550025, China
3. College of Science, Jiamusi University, Jiamusi 154007, China
4. China-Ukraine International Joint Laboratory for Agricultural and Forestry Technology Development and Application, Jiamusi University, Jiamusi 154007,China

Received:2019-02-27 Accepted:2020-11-26 Online:2021-03-30 Published:2021-04-09
Contact: DUAN Wenbiao E-mail:feimen5633525@126.com;dwbiao88@163.com

Abstract

Abstract:

【Objective】The community structure and diversity characteristics of soil nirK-type denitrifying microorganisms in three types of primitive Pinus koraiensis forest (Picea asperata-Abies fabri-Pinus koraiensis forest, Tilia spp.-P. koraiensis forest, and Betula costata-P. koraiensis forest) were analyzed, and the key soil physicochemical factors affecting the soil nirK-type denitrifying microbial community were explored in the Liangshui National Nature Reserve of the Lesser Khingan Mountains.【Method】Soil samples of the selected three types of primitive P. koraiensis forest were collected in the Liangshui National Nature Reserve of the Lesser Khingan Mountains of Yichun City, Heilongjiang Province. Additionally, total soil microbial DNA was extracted, and variable range fragments of the nirK gene, which encodes the coding gene of nitrate reductase (a key enzyme in the denitrifying process), were amplified with PCR. The community composition and diversity characteristics of soil nirK-type denitrifying microorganisms in the different forest types of P. koraiensis were analyzed by using high-throughput sequencing and bioinformatic analysis.【Result】A total of 761 912 high-quality sequences of the 848 312 effective sequences of the nirK gene were obtained with high-throughput sequencing of 12 soil samples in the three types of primitive P. koraiensis forest, and the average length of these sequences was 320 bp. Soil nirK-type denitrifying microorganisms of the three forest types belong to bacteria and archaea.In addition there were a large number of sequences without clear classification information. The soil nirK denitrifying microorganisms identified in the three forest types belonged primarily to Proteobacteria at the Phylum level. However, the core genera included Bradyrhizobium, Rhodopseudomonas, Azospirillum and Burkholderia. Alpha diversity analysis showed that there was no significant difference in the four alpha diversity indices (Shannon, Chao1, Ace and Simpson index) of soil nirK-type denitrifying microorganisms among the three forest types. Beta diversity analysis showed that there were significant differences in the composition of soil nirK microbial communities among the three forest types (R=0.25, P=0.016). Metastats analysis showed that the significant differences in soil nirK microbial communities among the three forest types were with regards to Proteobacteria, Euryarchaeota and Bacteroidetes. Soil ammonium nitrogen and total nitrogen were the main soil physicochemical factors influencing soil nirK-type denitrifying microorganisms communities in the three forest types (P<0.05).【Conclusion】There was no significant difference in the alpha diversity index of the three primitive P. koraiensis forests (Shannon index, Simpson index, Ace index and Chao1 index), although beta diversity analysis showed that there were significant differences in the composition of the soil nirK microbial community among the three types of primitive P. koraiensis forest (R=0.25, P=0.016). Ammonium nitrogen and total nitrogen were the main soil physicochemical factors that significantly affected the soil nirK denitrifying microorganism in the three forest types.

Key words: Lesser Khingan Mountains, primitive Pinus koraiensis forest, nirK gene, denitrifying microorganisms, community structure, high-throughput sequencing, Liangshui National Nature Reserve

CLC Number:

S718.81

CHEN Xiubo, DUAN Wenbiao, CHEN Lixin, ZHU Dequan, ZHAO Chenchen, LIU Dongxu. Community structure and diversity of soil nirK-type denitrifying microorganisms in three forest types of primitive Pinus koraiensis mixed forest in Liangshui National Nature Reserve, Lesser Khingan Mountains[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2021, 45(2): 77-86.

Figures/Tables 8

Table 1

Table 2

Physicochemical properties of soil in three types of primitive Pinus koraiensis mixed forest of Lesser Khingan Mountains"

林型 forest type	pH	土壤含水量/% soil water content	总有机碳/ (g·kg^-1) total organic carbon	全氮/ (g·kg^-1) total nitrogen	全磷/ (mg·kg^-1) total phosphorus	总孔隙度/% total porosity	土壤容重/ (g·cm^-3) soil bulk density	$NO 3 -$ -N含量/ (mg·kg^-1) content of $NO 3 -$ -N	N $H 4 +$ -N/ 含量 (mg·kg^-1) content of $NH 4 +$ -N
A	6.13±0.15 a	62.79±4.18 a	213.69±21.28 a	4.48±0.18 a	5.46±0.35 b	46.46±2.27 b	0.89±0.06 a	5.13±0.25 a	20.36±0.95 c
B	6.11±0.25 a	78.21±7.32 b	216.32±20.43 a	4.69±0.07 a	5.17±0.29 b	51.86±1.63 a	0.93±0.06 a	4.76±0.36 b	25.00±1.63 b
C	5.95±0.56 a	62.46±3.36 a	202.80±36.54 a	6.85±0.92 b	9.87±0.54 a	51.68±1.91 a	0.93±0.02 a	4.26±0.16 c	36.15±3.10 a

Table 2

Fig.1

Fig.2

Table 3

Fig.3

Fig.4

Fig.5

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林型 forest type	林分密度/ (株·hm^-2) stand density	平均树高/m mean height	平均胸径/cm mean DBH	郁闭度/% canopy	主要植物 main plants
云冷杉红松林 Picea asperata and Abies fabri- Piuns koraiensis forest	156.00±40.00	25.27±1.51	50.33±4.25	81.50±4.51	冷杉(Abies fabri)、云杉(Picea asperata)、花楷槭(Acer ukurunduense)、青楷槭(A. tegmentosum)、五角槭(A. mono)
椴树红松林 Tilia spp.-Pinus koraiensis forest	128.00±39.19	26.37±1.67	54.45±6.26	84.50±5.45	槺椴(Tilia mandschurica)、紫椴(T. amurensis)、花楷槭(A. ukurunduense)、青楷槭(A. tegmentosum)、五角槭(A. mono)
枫桦红松林 Betula costata- Pinus koraiensis forest	128.00±42.10	28.60±2.60	59.80±8.40	89.50±6.30	枫桦(Betula costata)、五角槭(A. mono)、臭松(Abies nephrolepis)、稠李(Prunus padus)、青楷槭(A. tegmentosum)、椴树(Tilia spp.)、毛榛子(Corylus mandshurica)、瘤枝卫矛(Euonymus verrucosus)等

林型 forest types	Shannon 指数 Shannon index	Ace指数 Ace index	Chao1 指数 Chao1 index	Simpson 指数 Simpson index
云冷杉红松林 Picea asperata and Abies fabri- Pinus koraiensis forest	8.22± 0.18 a	1433± 329.86 a	1414± 303.83 a	0.99± 0.003 a
椴树红松林 Tilia spp.- P. koraiensis forest	7.79± 2.19 a	1844± 532.61 a	1779± 506.67 a	0.90± 0.16 a
枫桦红松林 Betula costata- P. koraiensis forest	7.45± 1.25 a	1630± 915.70 a	1578± 836.18 a	0.93± 0.04 a

Community structure and diversity of soil nirK-type denitrifying microorganisms in three forest types of primitive Pinus koraiensis mixed forest in Liangshui National Nature Reserve, Lesser Khingan Mountains

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