Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (5): 980-991.doi: 10.3864/j.issn.0578-1752.2021.05.010

• SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT • Previous Articles     Next Articles

Effects of Straw Addition on Soil Biological N2-Fixation Rate and Diazotroph Community Properties

Xu LI(),WeiLing DONG,ALin SONG,YanLing LI,YuQiu LU,EnZhao WANG,XiongDuo LIU,Meng WANG,FenLiang FAN()   

  1. Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences/Key Laboratory of Plant Nutrition and Fertilizer, Ministry of Agriculture and Rural Affairs, Beijing 100081
  • Received:2020-06-21 Accepted:2020-12-16 Online:2021-03-01 Published:2021-03-09
  • Contact: FenLiang FAN E-mail:82101185083@caas.cn;fanfenliang@caas.cn

Abstract:

【Objective】The purpose of this study was to analyze the effects of different straw additions on soil N2-fixation rate and diazotroph community structure, which was crucial for the management of crop residue and mineral fertilizer application in China.【Method】Using indoor incubation experiment, in addition to the control (C0: 0), a total of 5 straw addition gradients (C1: 0.2 mg·g-1; C2: 1.0 mg·g-1; C3: 2.0 mg·g-1; C4: 4.0 mg·g-1; C5: 10.0 mg·g-1), using the 15N2 labeling method, and soil samples were collected after 28 days of incubation in dark conditions. As a direct measure of N2-fixation,15N2 gas labeling method could quantify the rate of biological N2-fixation, and destined for characterization of nifH gene marker and diazotroph community by using the Illumina PE250 sequencing and PCR techniques. 【Result】With the increase of straw addition, the content of NO3--N in soil decreased significantly, while the content of NH4+-N did not change significantly. The pH of soil decreased and the rate of biological N2-fixation increased significantly. C3, C4 and C5 treatments could reach 2.71-3.05 μg N·g-1DW during the incubation period (28 d), which was about 87-96 kg N·hm-2·a-1, compared with C0, the rate of biological N2-fixation increased by 38.1%-52.4%. The copy number of nifH gene ranged from 5.48×107 to 9.20×107 copies/(g soil) under all treatments. Compared with C0, the copy number of nifH gene was significantly increased under C4 and C5 treatments, and the number and activity of diazotroph were significantly increased (P<0.05). However, when the amount of straw added reached 4.0 mg·g -1, increasing the amount of straw had no significant effect on the copy number of nifH gene of soil diazotroph. With the increase of straw addition, Shannon-Weill index of C4 and C5 levels was significantly lower than that of the other four treatments (P<0.05), and the diversity of the other four treatments had no significant difference. The result of PCoA showed that diazotroph community structure was clustered into different groups depending upon the difference of straw addition. Diazotroph were divided into Proteobacteria, Cyanobacteria, Firmicutes, Actinobacteria and Spirochaetes at the phylum level. As the amount of straw added increases, the relative abundance of Cyanobacteria tends to increase first and then decrease. At the genus level, there were significant differences in the number of carbon sources among different species. Bradyrhizobium was the most abundant genus. With the increase of straw addition, the relative abundance of C5 was significantly different from that of C0, C1, C2 and C3 (P<0.05), increasing by 12.07%-14.13%. The relative abundance of Scytonema hofmanni, which was positively correlated with the rate of biological N2-fixation, gradually increases with the increase of straw addition, but decreased when straw addition reaches C5 level (P<0.05). Meanwhile, PLS-PM analysis, RDA and correlation analysis revealed that the rate of biological N2-fixation and soil diazotroph community were greatly affected by straw addition and soil NO3--N concentration. 【Conclusion】The rate of soil biological N2-fixation increases with the amount of straw added. The effect of straw on N2-fixation was mainly due to the reduction of NO3--N content in the soil by straw addition, which in turn promoted the diazotroph such as Bradyrhizobium, Scytonema hofmanni and Azospirillum grow. According to the calculation results of this experiment, compared with no straw, after the straw was added (4.0 mg·g -1), the rate of biological N2-fixation could be increased by 26 kg N·hm-2·a-1, which significantly reduced the fertilizer demand.

Key words: soil diazotroph, straw returning, the rate of biological N2-fixation, 15N2 labeling method, nifH

Table 1

The physical and chemical properties and biological N2-fixation of soil different treatment"

处理
Treatment
pH NO3--N
(mg·kg-1)
NH4+-N
(mg·kg-1)
TN
(g·kg-1)
生物固氮量
Biological N2-fixation (μg N·g-1DW)
C0 7.07±0.01ab 15.33±0.26a 11.81±1.07a 1.18±0.01e 2.00±0.18d
C1 7.10±0.01a 14.44±0.14b 11.98±0.93a 1.21±0.00cd 2.25±0.05cd
C2 7.03±0.02b 13.80±0.09c 11.53±0.77a 1.20±0.01de 2.50±0.11bc
C3 7.06±0.03ab 11.25±0.14d 11.77±0.66a 1.24±0.01b 2.78±0.10ab
C4 6.96±0.03c 5.97±0.30e 11.13±0.14a 1.23±0.00bc 2.71±0.15ab
C5 6.93±0.03c 3.46±0.24f 11.85±0.81a 1.28±0.00a 3.05±0.13a

Fig. 1

Diazotroph community structure assessed by principal coordinate analysis (A), nifH gene copy number (B) and Shannon-Wiener index (C) of soil diazotroph (>1%); Relative abundance of phylum (%) (D), relative abundance (%) of most abundant genera (>1%) (E) under different straw additions Values followed by different letters means significant differences in different straw additions (P<0.05), * Mean P<0.05"

Table 2

The correlations between environmental factors and abundance, diversity and N2-fixation rate of soil diazotroph community"

nifH基因拷贝数
nifH copy numbers
固氮速率
N2-fixation rate
香农-威尔指数
Shannon-Wiener index
R P R P R P
秸秆添加Straw addition 0.708 0.000 0.804 0.000 -0.634 0.002
NO3--N -0.737 0.000 -0.828 0.000 0.621 0.002
NH4+-N -0.166 0.508 0.019 0.929 0.054 0.851
TN 0.483 0.027 0.598 0.003 -0.340 0.144
pH -0.623 0.002 -0.410 0.073 0.678 0.001

Fig. 2

Redundancy analysis (A) of the diazotroph community structure affected by physicochemical variables and PLS-PM analysis (B) ***, **, * mean P<0.001, P<0.01, P<0.05 respectively. The same as Fig.3"

Fig. 3

Spearman correlation analysis between relative abundance of diazotroph genera and soil physicochemical variables and N2-fixation rate"

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