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Frontiers of Agricultural Science and Engineering

ISSN 2095-7505

ISSN 2095-977X(Online)

CN 10-1204/S

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Front. Agr. Sci. Eng.    2019, Vol. 6 Issue (2) : 162-171    https://doi.org/10.15302/J-FASE-2018212
RESEARCH ARTICLE
The diazotrophic community in oat rhizosphere: effects of legume intercropping and crop growth stage
Yadong YANG, Xiaomin FENG, Yuegao HU, Zhaohai ZENG()
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China
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Abstract

In this study, the abundance, diversity and structure of the diazotrophic community in oat rhizosphere soil in three cropping systems and at two oat growth stages were investigated using real-time PCR and Illumina MiSeq sequencing. The nifH gene abundance in oat-soybean intercropping (OSO) and oat-mungbean intercropping (OMO) was significantly greater than that in sole oat (O), but the nifH gene abundance significantly decreased at the later stage in all the treatments. Alpha diversity indices in OSO and OMO were higher at the heading stage, but lower at the maturity stage than that in O. Bradyrhizobium and Skermanella were the dominant genera identified in all samples, with an average proportion of 35.8% and 12.4%, respectively. The proportion of dominant genera showed significant differences and varied with cropping system and growth stage. Principal component analysis showed that growth stage had a stronger effect than intercropping on the diazotrophic community structure. However, Mantel test and redundancy analysis showed there was no environmental factor significantly correlated to the diazotrophic community structure. Our results demonstrate that intercropping had a weaker effect than growth stage on the abundance, diversity and structure of the diazotrophic community in oat rhizosphere soil.
Keywords community composition      Illumina MiSeq sequencing      nifH gene      oat-legume intercropping      rhizosphere soil     
Corresponding Author(s): Zhaohai ZENG   
Just Accepted Date: 02 February 2018   Online First Date: 03 April 2018    Issue Date: 22 May 2019
 Cite this article:   
Yadong YANG,Xiaomin FENG,Yuegao HU, et al. The diazotrophic community in oat rhizosphere: effects of legume intercropping and crop growth stage[J]. Front. Agr. Sci. Eng. , 2019, 6(2): 162-171.
 URL:  
https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2018212
https://academic.hep.com.cn/fase/EN/Y2019/V6/I2/162
Sample pH (H2O) TN /(g·kg−1) NH4+-N /(mg·kg−1) NO3−-N /(mg·kg−1) TOM /(g·kg−1) C/N ratio
O_H 7.30±0.01 c 1.02±0.06 a 0.94±0.11 b 1.51±0.08 e 17.30±0.83 a 9.84±0.09 b
OSO_H 7.12±0.01 d 1.00±0.01 ab 1.24±0.07 a 5.25±0.39 b 17.21±0.95 a 9.96±0.49 b
OMO_H 7.00±0.02 d 0.96±0.01 bc 1.00±0.08 b 3.09±0.24 c 17.93±0.61 a 10.86±0.33 a
O_M 7.84±0.01 a 0.95±0.02 c 0.73±0.05 c 2.19±0.18 d 17.95±0.77 a 10.99±0.27 a
OSO_M 7.74±0.02 b 0.97±0.02 abc 0.96±0.08 b 10.62±0.53 a 18.50±0.86 a 11.03±0.32 a
OMO_M 7.85±0.02 a 0.95±0.01 bc 0.12±0.05 d 1.45±0.49 e 18.01±0.49 a 10.95±0.28 a
Tab.1  Chemical properties of the rhizosphere soil samples
Fig.1  Abundance of nifH gene in different rhizosphere soil samples. O_H, OSO_H, OMO_H, O_M, OSO_M and OMO_M refer to sole oat, oat-soybean intercropping and oat-mungbean intercropping at the heading and maturity stages of oat, respectively. The nifH gene copy numbers were log-transformed. Values are mean±SD (n = 3). Different letters indicate significant differences (P<0.05).
Sample Sequencing results Diversity estimates (97%)
Reads OTUs ACE Chao1 Shannon
O_H 7168 104 119.97 (110.83, 141.32) 116.67 (108.32, 141.16) 2.43 (2.39, 2.46)
OSO_H 7168 139 143.64 (140.49, 153.45) 145.60 (140.71, 164.43) 2.76 (2.72, 2.80)
OMO_H 7168 116 127.04 (121.51, 141.33) 125.06 (120.10, 141.67) 2.64 (2.61, 2.68)
O_M 7168 125 132.08 (128.07, 143.82) 129.88 (126.95, 141.89) 3.10 (3.07, 3.14)
OSO_M 7168 108 116.82 (111.28, 131.74) 117.55 (110.82, 140.34) 2.88 (2.85, 2.92)
OMO_M 7168 99 117.47 (108.00, 139.77) 122.00 (108.30, 161.45) 2.65 (2.62, 2.69)
Tab.2  MiSeq sequencing results and diversity estimates of different rhizosphere soil samples
Fig.2  Taxonomic distribution of the diazotrophic genera in different rhizosphere soil samples. O_H, OSO_H, OMO_H, O_M, OSO_M and OMO_M refer to sole oat, oat-soybean intercropping and oat-mungbean intercropping at the heading and maturity stages of oat, respectively.
Group pH TOM TN NH4+-N NO3−-N C/N ratio
Skermanella 0.825* 0.848* 0.805 0.452 0.287 0.886*
Bradyrhizobium 0.840* 0.620 0.652 0.945** 0.190 0.671
Proteobacteria_unclassified 0.910* 0.779 0.771 0.604 0.114 0.832*
Bacteria_unclassified 0.219 0.457 0.767 0.170 0.259 0.650
Azohydromonas 0.437 0.737 0.876* 0.186 0.220 0.884*
Alphaproteobacteria_unclassified 0.582 0.188 0.060 0.792 0.347 0.118
Rhizobiales_unclassified 0.486 0.085 0.072 0.183 0.312 0.086
Tab.3  Pearson correlation coefficients between rhizosphere soil properties and the abundance of diazotrophic groups (average proportion>1%) at the genus level
Fig.3  Redundancy analysis (RDA) displaying the relationship between rhizosphere soil properties, rhizosphere soil samples and relative abundance of the diazotrophic genera. Only the known genera are shown on the RDA plot. O_H, OSO_H, OMO_H, O_M, OSO_M and OMO_M refer to sole oat, oat-soybean intercropping and oat-mungbean intercropping at the heading and maturity stages of oat, respectively.
Soil property RDA1 RDA2 r2 Pr (>r)
pH ??0.61829 ??0.78595 0.0722 0.8708
TOM −0.98959 −0.14389 0.1216 0.8208
TN −0.98811 ??0.15372 0.0895 0.8667
NH4+-N −0.92327 −0.38416 0.0747 0.8986
NO3--N −0.80804 −0.58913 0.3965 0.4778
Tab.4  Significance of rhizosphere soil property variables in explaining the diazotrophic community structures observed from the RDA results
Fig.4  Principal component (PC) analysis displaying the diazotrophic community structure at the genus level. O_H, OSO_H, OMO_H, O_M, OSO_M and OMO_M refer to sole oat, oat-soybean intercropping and oat-mungbean intercropping at the heading and maturity stages of oat, respectively.
Fig.5  Venn drawing showing the diazotrophic genera detected at the heading (a) and maturity (b) stages of oat in different rhizosphere soil samples. Shared genera are shown in the overlap parts of the circles. O_H, OSO_H, OMO_H, O_M, OSO_M and OMO_M refer to sole oat, oat-soybean intercropping and oat-mungbean intercropping at the heading and maturity stages of oat, respectively.
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