长期施肥对棕壤丛枝菌根真菌群落结构及其侵染的影响

doi:10.3864/j.issn.0578-1752.2018.24.008

Abstract

Abstract:

【Objective】AM (Arbuscular mycorrhizal) fungi plays important roles like improving the rhizosphere soil environment, promoting the uptake of nutrients by plants, enhancing resistance of plant, and increasing crop yield and quality. This study aimed to explore changes of community structures and colonization rate of AM fungi and to find out the main factors which affected the changes under a corn-soybean rotation system and a long-term fertilization for 38 years in a brown soil. 【Method】 Soil samples (0-20 cm) were taken from the six treatments of the long-term fertilization experiment in June 2016, including (1) no fertilizer (CK); (2) chemical N input (N); (3) chemical N and P input (NP); (4) chemical N, P and K input (NPK); (5) pig manure (M); (6) pig manure, chemical N and P (MNP). Then the soil samples were analyzed by using PCR-DGGE, gel-recovery, sequencing and trypan blue staining. Relationship between community and colonization rate of AM fungi and environmental factors were analyzed by Redundancy analysis and Canonical Correlation analysis. 【Result】 The result showed that the contents of alkali-hydrolysable nitrogen (AHN), available phosphorus (AP), available potassium (AK) , ammonium nitrogen (NH₄ ⁺-N), nitric nitrogen (NO₃ ^--N) and dissolved organic carbon (DOC) under organic fertilization treatments were significantly higher than them under the CK treatment and chemical fertilization treatments, and the trend was: organic fertilizer treatments > chemical fertilizer treatments > CK treatment. Compared with the CK treatment, soil pH was decreased in chemical fertilizer treatments and increased in organic fertilizer treatments. 22 bands of AM fungi from soil and 9 bands of AM fungi from root were obtained by gel-recovery, and 13 OTU were obtained by BLAST. The result of sequencing showed that AM fungi species isolated from soil samples were mainly Glomeromycota and Gigasporaceae, while infected AM fungi was only Glomeromycota. The cluster analysis showed that community structures of soil AM fungi were divided into three groups under a long-term fertilization in a brown soil, namely N treatment, organic fertilizer treatments and another fertilizer treatments. Community structures of infected AM fungi were also divided into three groups, namely NPK treatment, M treatment and NP treatment, and another fertilizer treatments. The spore density of AM fungi under organic fertilizer treatments was significantly higher than that under chemical fertilizer treatments and non-fertilizer treatment, and the trend was: organic fertilizer treatments > chemical fertilizer treatments > CK treatment. The trend of colonization rate of AM fungi under different fertilizer treatments was: NPK treatment > organic fertilizer treatments > another fertilizer treatments. Redundancy analysis showed that spore density was positively correlated with soil AHN, NH₄ ⁺-N, AP, AK, DOC and soil moisture content, and colonization rate was positively correlated with nitric nitrogen content. The colonization rate was positively correlated with spore density, while diversity index of AM fungi was neither correlation with colonization rate nor with spore density. Canonical Correlation analysis showed AHN, AK, DOC and NH₄ ⁺-N significantly influenced the ribotypes of AM fungi. 【Conclusion】 The long-term fertilization changed community structures of AM fungi by changing the physicochemical properties of the soil, and then affected colonization of AM fungi.

Key words: Arbuscular mycorrhizal fungi, brown soil, long-term fertilization, community structure, colonization rate

GUO Jing,LUO PeiYu,YANG JinFeng,LI DongDong,HUANG YueYue,HAN XiaoRi. Influence of Long-term Fertilization on Community Structures and Colonization of Arbuscular mycorrhizal Fungi in a Brown Soil[J].Scientia Agricultura Sinica, 2018, 51(24): 4677-4689.

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引物名称Primer	引物序列（5′-3′）Primer sequence（5′-3′）	反应条件Conditions
NS1	GTAGTCATATGCTTGTCTC	94℃预变性3 min;94℃ 1 min,50 ℃ 1 min,72 ℃ 1 min,30个循环;72 ℃ 7 min (一轮) An initial denaturation of 94℃for 3 min; 94℃ 1 min, 50℃ 1 min, 72℃ 1 min, 30 cycles; 72℃ 7 min (The first round)
NS41	CCCGTGTTGAGTCAAATTA
GC-NS31	TTGGAGGGCAAGTCTGGTGCC
AM1	GTTTCCCGTAAGGCGCCGAA	94 ℃预变性3 min;94 ℃ 30 s,58 ℃ 45 s,72 ℃ 45 s,28个循环;72 ℃ 7 min（二轮） An initial denaturation of 94℃ for 3 min; 94℃ 30 s, 58℃ 45 s, 72℃ 45 s, 28 cycles; 72℃ 7 min (The second round)
GC-ARCH1311	TGCTAAATAGCCAGGCTGY
NS8	TCCGCAGGTTCACCTACGGA

处理 Treatment	pH (H₂O)	速效磷 AP (mg·kg^-1)	速效钾 AK (mg·kg^-1)	碱解氮 AHN (mg·kg^-1)	硝态氮 NO₃^--N (mg·kg^-1)	铵态氮 NH₄⁺-N (mg·kg^-1)	可溶性有机碳 DOC (mg·kg^-1)	土壤含水量 SM (%)
CK	5.75±0.01b	1.19±0.64d	96.66±1.21c	85.04±0.68f	12.06±0.02e	3.97±0.18d	39.42±0.26e	15.40±0.14b
N	4.87±0.00d	2.14±0.64d	81.62±1.86c	110.43±0.94d	48.38±1.03c	4.45±0.12cd	65.41±0.05d	15.41±0.22b
NP	5.35±0.01c	16.15±0.22c	76.94±2.32c	96.60±1.17e	51.22±1.39bc	4.83±0.16bc	71.29±0.74c	15.12±0.38b
NPK	4.99±0.01c	17.10±0.63c	107.02±1.16c	120.73±2.43c	55.97±0.74b	5.26±0.046b	73.10±0.17c	15.80±0.04b
M	6.64±0.06a	137.72±2.29b	209.96±9.98b	132.54±1.74b	41.81±0.75d	5.33±0.09b	108.44±2.51b	18.11±0.04a
MNP	5.78±0.02b	182.15±7.01a	268.12±9.75a	165.56±2.24a	73.63±0.43a	7.75±0.12a	126.45±1.30a	18.20±0.73a

处理 Treatment	香农指数 Shannon-Wiener(H)	均匀度 Evenness (E)	条带数 Bands
CK	2.04±0.01c	0.982±0.00a	8
N	1.93±0.01d	0.988±0.01a	7
NP	2.18±0.02a	0.991±0.01a	9
NPK	1.78±0.03e	0.997±0.02a	6
M	2.09±0.05b	0.999±0.01a	8
MNP	1.78±0.02e	0.995±0.01a	6

处理 Treatment	香农指数 Shannon-Wiener(H)	均匀度 Evenness(E)	条带数 Bands
CK	0.88±0.06c	0.799±0.02b	3
N	1.03±0.09c	0.740±0.01b	4
NP	1.91±0.03a	0.981±0.01a	7
NPK	1.92±0.02a	0.985±0.01a	7
M	1.91±0.01a	0.984±0.01a	7
MNP	1.36±0.01b	0.988±0.01a	4

Influence of Long-term Fertilization on Community Structures and Colonization of Arbuscular mycorrhizal Fungi in a Brown Soil

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