ACTA AGRONOMICA SINICA

Volume 37, Issue 1, January 2011

Online English edition of the Chinese language journal

Cite this article as: Acta Agron Sin, 2011, 37(1): 105–111. RESEARCH PAPER

Effect of Different Fertilizers on Functional Diversity of

Microbial Flora in Rhizospheric Soil Under Tobacco
Monoculture

YANG Yu-Hong1, CHEN Dong-Mei2, JIN Yan1, WANG Hai-Bin2, DUAN Yu-Qi1, GUO Xu-Kui2,
HE Hai-Bin2, and LIN Wen-Xiong2,*
1
Yunnan Provincial Institute of Tobacco Research, Yuxi 653100, China
2
Institute of Agroecology, Fujian Agriculture and Forestry University, Fuzhou 350002, China

Abstract: Continuous cropping obstacle is one the most important problems in tobacco (Nicotiana tabacum L.) production. To alleviate
continuous cropping obstacle based on management of soil ecology, the effects of different fertilizers were tested using tobacco cultivar
K326 growing in a field with 12-year consecutive cultivation. The rhizospheric soil was sampled to investigate the changes in functional
diversity of microbial flora in different treatments. The results showed that the autotoxic allelopathic potential was maximal for the
monoculture soil treated with traditional compound fertilizer, and minimal for the soil treated with farmyard manure. According to the
result of BIOLOG analysis, traditional compound fertilizer was conducive to the growth of microbial flora feeding on amino acids and
amine as carbon sources, the commercial organic fertilizer to the growth of microbial flora using carboxylic acids as a carbon source, and
farmyard manure to the growth of microbial flora using carbohydrate, fatty acids, and phenolic acids as carbon sources. Principal
component analysis indicated that the first 2 components were related to carbon sources, which accounted for 74.37% and 25.63% of the
data variation. The carbon source of carbohydrate, fatty acids, and phenolic acids mainly contributed to the separation of the 2 principal
components. The autotoxic allelopathic potential of tobacco rhizospheric soil was positively correlated with the average well color
development (AWCD) value of microbial flora feeding on carbohydrate and phenolic acids as carbon sources, and negatively correlated
with that of the microbial flora using the carbon source of fatty acids. In addition, for the growth of microbial flora in monoculture soil,
farmyard manure was the best, followed by commercial organic fertilizer, and traditional compound fertilizer was the worst.

Keywords: tobacco; consecutive cropping problem; microbial flora; functional diversity; BIOLOG

Continuous cropping results in serious reduction of crop
biomass and quality, and increases the susceptibility of crop
disease, which is known as continuous cropping obstacles.
Tobacco (Nicotiana tabacum L.), a member of Solanaceae, is
one of the important economic crops with high economic
benefits and widely planted in the world [1]. However,
continuous cropping obstacles always seriously restrict the
sustainable development of tobacco production. The
mechanism of continuous cropping obstacles is highly
concerned because the problem becomes greater with the
cropping years.
Although no consistent conclusion is available regarding
the mechanism of continuous cropping obstacles, a general
understanding points to several factors involved in the
mechanism, such as imbalance of soil nutrients, autotoxic
effect, and changing of soil microbial community [2–5]. These
factors are all related to soil ecosystem. The physical and
chemical processes in soil are very complex, and micro-
organisms play a crucial role in the processes, particularly in
soil nutrient transformation, soil health, and sustainability of
soil productivity. Pan et al. [6] found that continuous cropping
tobacco affected the communities of bacteria, fungi, and
actinomycosis in soil with the effect from large to small.
Tobacco continuous cropping disturbed the balance of

Received: 15 June 2010; Accepted: 26 September 2010.

* Corresponding author. E-mail: wenxionglin@163.com
Copyright © 2011, Crop Science Society of China and Institute of Crop Sciences, Chinese Academy of Agricultural Sciences. Published by Elsevier BV. All rights reserved.
Chinese edition available online at http://www.chinacrops.org/zwxb/
DOI: 10.1016/S1875-2780(11)60003-5
 YANG Yu-Hong et al. / Acta Agronomica Sinica, 2011, 37(1): 105–111
microbial communities and increased disease susceptibility of
the crop [7]. Reasonable fertilization could optimize the
structure of microbial communities in rhizospheric soil of
tobacco in continuous cropping system, improve the microbial
diversity, and alleviate continuous cropping obstacles [8–10].
Therefore, continuous cropping obstacles are considered to be
closely related to soil fertility and microbial community. At
present, numerous studies have been carried out to understand
the characteristics of rhizosperic microbial florae in different
crops or soil types, and the effects of different fertilizers on
microbial flora in rhizospheric soil were reported in
monoculture tobacco [11–13]. However, the quantitative changes
of microorganisms in soils were emphasized in these studies,
and the functional diversity of microbial flora was seldom
mentioned. In the current study, we investigated the effects of
different fertilizer types on functional diversity of microbial
communities in a field with 12-year continuous cropping of
tobacco. The result was expected to help understanding the
mechanism of continuous cropping obstacles and provide
feasible techniques for alleviating this problem in tobacco
production.
1 Materials and methods
1.1 Experimental design
The field experiment was conducted in Yuxi Experiment
Base of Yunnan Tobacco Research Institute (Yuxi, Yunnan
Province, China) from 2008 to 2009. The tobacco accession
K326 was planted in the field where tobacco has been grown
continuously for 12 years. The nutrient basis of soil (pH 7.6)
before planting was as follows: organic matter 2.2%, total
nitrogen 0.95 g kg1, total phosphorus 1.70 g kg1, total
potassium 10.0 g kg1, available nitrogen 45.9 mg kg1,
available phosphorus, 69.1 mg kg1, and available potassium
160.5 mg kg1. The cation exchange capacity was 4.44 cmol
kg1.
Three treatments were designed with different fertilizer
types, i.e., traditional compound fertilizer (T1, control),
traditional compound fertilizer plus farmyard manure (T2),
and traditional compound fertilizer plus commercial organic
fertilizer (T3). The farmyard manure was the pig manure
deposited for 3 months (nitrogen content of 0.4%). All
treatments had the same ratio of nutrient components as that
of the traditional compound fertilizer, which was 121224 for
NP2O5K2O, and the amount of nitrogen applied in each
treatment was about 75 kg ha1. Each treatment had 3
replicates, and the whole experiment contained 9 plots. The
plot area was 28 m2.
Tobacco seedlings were transplanted on April 25, and the
density was 56 seedlings per plot. A half of traditional
compound fertilizer was used as the basal fertilizer on April
26, which was placed 10 cm distant from the hill and covered
with 10–15 cm soil; another 10 g per plant was given with
irrigation water on May 5; and the remaining fertilizer was
applied on May 25. The farmyard manure in T2 treatment was
broadcasted before planting at the rate of 1 kg per plant. The
rest of fertilizer was applied in the same method as mentioned
above (T1 and T3). After harvest, the rhizospheric soils of
tobacco were collected using quartering method and preserved
at 4 qC.
1.2 Toxic effect of tobacco rhizosphere soil from
continuous cropping system
The toxic effects of tobacco rhizospheric soil on lettuce
(Lactuca sativa L.) and tobacco were estimated using the
soil-agar sandwich method [14]. Fifteen grams of the soil
sample were mixed with 30 mL agarose (0.8%) when the
agarose temperature was about 45qC. After solidification of
the agarose, extra 2 mL of agarose (0.5%) was added onto the
surface. Then, 10 pre-germinated seeds of lettuce or tobacco
were placed onto the agarose surface for germination and
growth. The cultural medium without rhizospheric soil was
used as the control. Each treatment had 5 replicates. The
lettuce seeds were incubated for 3 d under 25qC and 12 h
photoperiod, and the tobacco seeds were under 30qC and 12 h
photoperiod for 10 d. The root lengths of lettuce and tobacco
were measured. The relative inhibition rate (IR) was
calculated using the formula IR = [(value of control  value of
treatment) / value of control] × 100%.
1.3 Functional diversity of microbial community in
tobacco rhizospheric soil
The BIOLOG ECO microplate was used to determine the
microbial function diversity of soil [14]. Five grams of
freshly-collected soil was put into an autoclaved flask with
100 mL NaCl (0.85%). The flask was shaken at 120 rpm for 3
min, and then cooled on ice for 2 min. Subsequently, 5 mL of
supernatant was transferred into another flask with 45 mL
sterile distilled water. After 3 times of dilution, the 1:1000
extract was used for enzyme-linked immunosorbent assay
(ELISA). An aliquot of 150 ȝL extract solution was added
into each well of a BIOLOG ECO plate that was prewarmed
to 25 qC. The plate was incubated at 28 qC, and the absorbance
at 590 nm was recorded everyday until the seventh day using
the ELISA reaction plate reader (Multiscan MK3, Thermo
Fisher Scientific, MA, USA). Each well of the BIOLOG ECO
plate (Biolog, Hayward, CA, USA) was loaded with a
single-carbon source. Thirty-one single-carbon sources in 6
classes were used in the analysis, such as carbohydrate, fatty
acid, amino acid, amine, carboxylic acid, and phenolic acid.
ELISA reaction of microorganisms was presented as the
average well color development (AWCD) for each microplate,
which was calculated using the formula AWCD = [™(C  R)] /
N, where C is the absorbance of the 31 carbon source wells, R
 YANG Yu-Hong et al. / Acta Agronomica Sinica, 2011, 37(1): 105–111
is the absorbance of the corresponding control wells, and N
refers to the number of carbon source.
1.4 Data analysis
Statistical analysis, principal component analysis (PCA), and
correlation analysis were conducted using SSPS 11.5 software.
2 Results
2.1 Toxic effect of tobacco rhizospheric soil from
continuous cropping system
Compared to the control, the root lengths of lettuce and
tobacco seedlings were significantly inhibited in the
rhizospheric soil of continuous cropping tobacco, and the
inhibitory effect (IR) varied with the fertilizer supply (Fig. 1).
For lettuce, the IRs were 45.15%, 18.03%, and 21.70% in
treatments T1, T2, and T3, respectively; for tobacco, the IRs
were 57.52%, 33.18%, and 37.81%, respectively. The result
showed that the autotoxic effect of tobacco in the continuous
cropping system was regulated by different fertilizers, and
farmyard manure had the best effect, whereas traditional
compound fertilizer had the worst effect. In addition, the
autotoxic effect of tobacco rhizospheric soil on lettuce
seedlings was greater than that on tobacco seedlings.
2.2 Functional diversity of microbial community in
rhizospheric soil under tobacco monoculture
The AWCD values of microflora increased with cultural
time regardless their different feeding carbon sources. The
microflora feeding on different carbon resources had signifi-
80
Tobacco
a
Inhibition rate (%)
60
a
40
b
c (fatty acid) and negative (phenolic acid) correlations (Table 1).
20
0 acid, and phenolic acid, which resulted in the differentiation
T1 T2 T3 T1
Treatment
Fig. 1 Autotoxic effect of soil mediated by continuous
cropping tobacco on root lengths of tobacco and
lettuce under different fertilization treatments
T1: Traditional compound fertilizer; T2: Traditional compound
fertilizer plus farmyard manure; T3: Traditional compound
fertilizer plus commercial organic fertilizer.
Error bar shows the derivation of 5 replicates.
Different letters above the bars indicate significant
difference (P < 0.05) among treatments.
cantly different AWCD values in the 3 treatments. When
carbohydrate, fatty acid, or phenolic acid was used as the
carbon resource, treatment T2 exhibited the largest effects on
AWCD values with the simulative equation gradients of 0.320,
0.289, and 0.222, respectively; comparatively, the gradients of
AWCD simulative equations were 0.318, 0.272, and 0.196 for
treatment T3 and of 0.293, 0.264, and 0.176 for treatment T1,
respectively. When amino acid was used as the carbon source,
the AWCD values of microflora were affected by fertilization
treatments in the order of T1 (0.365) > T3 (0.353) > T2
(0.344). For amide and carboxylic acid used as the carbon
sources, the effects on AWCD value by fertilizers showed the
orders of T1 (0.370) > T2 (0.360) > T3 (0.344) and T3 (0.373)
> T1 (0.332) > T2 (0.314), respectively (Fig. 2). Therefore,
the traditional compound fertilizer was conducive to the
growth of the microflora feeding on amino acids and amine as
carbon sources, and the commercial organic fertilizer and
farmyard manure were favorable for the microflora using
carboxylic acids and carbohydrate, fatty acids, and phenolic
acids as the carbon sources.
2.3 PCA of microflora in tobacco rhizospheric soil under
continuous cropping system
Based on AWCD values of microflora cultural for 7 d, the
functional diversity of microflora was analyzed using PCA
method. The utilization property of carbon source was
separated into 2 principal components, which explained
74.4% and 25.6% of the total variation. The soil samples from
T1, T2, and T3 treatments were clustered on different
positions in the PCA graph (Fig. 3). According to the
comparative analysis between PCA and AWCD value of
single-carbon source, the component 1 was significantly
correlated with 16 single-carbon sources, of which 12 had
positive correlations and 4 had negative correlation. The
Lettuce
positive carbon sources were mainly involved in carbohydrate
and phenolic acid, and the negative carbon sources were
b mainly in carbohydrate and fatty acid classes. The component
b
2 contained each one carbon source with significantly positive
Clearly, the major carbon sources were carbohydrate, fatty
T2 T3 of the principal components.
2.4 Correlation between microflora using different carbon
sources and autotoxic effect of tobacco rhizospheric soil
The correlation coefficient of autotoxic potential between
tobacco and lettuce was 1.00 (P < 0.01). On both tobacco and
lettuce in different fertilization treatments, the autotoxic effects
of rhizospperic soils from tobacco continuous cropping field
were positively correlated with the AWCD values of microflora
using carbohydrate and phenolic acid, but negatively correlated
with the AWCD values of microflora using fatty acid. In addi-
 YANG Yu-Hong et al. / Acta Agronomica Sinica, 2011, 37(1): 105–111

3 3 B
A

2 2

1 1

0 0
1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8

3 3
C D

2 2
AWCD value

1 1

0 0
1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8

3 3
E F

2 2
AWCD value

1 1

0 0
1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8

Days of microorganism culture Days of microorganism culture

T1 T2 T3

Fig. 2 Changes in averages of well color development (AWCD) values of tobacco rhizospheric microflora with different carbon sources
A: Carbohydrate; T1: y = 0.293x + 0.1544 (R2 = 0.8603), T2: y = 0.320x + 0.1493 (R2 = 0.8553), T3: y = 0.318x  0.0545 (R2 = 0.9611);
B: Fatty acid; T1: y = 0.264x + 0.1017 (R2 = 0.9196), T2: y = 0.289x  0.0052 (R2 = 0.9609), T3: y = 0.272x + 0.2061 (R2 = 0.8172);
C: Amino acid; T1: y = 0.365x  0.0948 (R2 = 0.9381), T2: y = 0.344x + 0.0907 (R2 = 0.8838), T3: y = 0.353x  0.0267 (R2 = 0.9422);
D: Amide; T1: y = 0.370x  0.0060 (R2 = 0.9340); T2: y = 0.360x + 0.0114 (R2 = 0.9055), T3: y = 0.344x + 0.0794 (R2 = 0.8809);
E: Carboxylic acid; T1: y = 0.332x  0.0087 (R2 = 0.8788), T2: y = 0.314x + 0.0732 (R2 = 0.9129), T3: y = 0.373x  0.0221 (R2 = 0.9256);
F: Phenolic acide; T1: y = 0.176x + 0.2972 (R2 = 0.7935), T2: y = 0.222x + 0.2284 (R2 = 0.7420), T3: y = 0.196x + 0.2209 (R2 = 0.7964).
T1: Traditional compound fertilizer; T2: Traditional compound fertilizer plus farmyard manure;
T3: Traditional compound fertilizer plus with commercial organic fertilizer.
 YANG Yu-Hong et al. / Acta Agronomica Sinica, 2011, 37(1): 105–111

Table 1 Carbon sources mainly correlated with components

of principal component analysis
Correlation
Carbon source Class of carbon source
coefficient
Principal component 1
Į-D-lactose Carbohydrate 1.00**
D-cellobiose Carbohydrate 1.00**
Glycogen Carbohydrate 1.00**
Į-cyclodextrin Carbohydrate 1.00**
D,L-Į-glycetol phosphate Carbohydrate 1.00**
Glucose-1-phosphate Carbohydrate 1.00**
D-galacturonic acid Carboxylic acid derivatives 1.00**
N-acetyl-D-glucosamine Amine 1.00**
D-mannitol Carbohydrate 1.00**
Fig. 3 Loadings for principal component analysis of tobacco
rhizospheric microflora under continuous cropping system D-xylose Carbohydrate 1.00**
T1: Traditional compound fertilizer; T2: Traditional compound ȕ-methyl-D-glucose Carbohydrate 1.00**
fertilizer plus farmyard manure; T3: Traditional compound 4-hydroxy benzoic acid Phenolic acid 0.98*
fertilizer plus with commercial organic fertilizer. Itaconic acid Carboxylic acid derivatives 1.00**
1-Erythritol Carbohydrate 1.00**
D-galacturonic acid Carboxylic acid derivatives 1.00**
tion, the AWCD values of microflora using phenolic acid was D-glucosaminic acid Amino acid 1.00**
negatively correlated with that using fatty acid (Table 2). This Principal component 2
result indicated that the autotoxic effect of tobacco Ȗ-hydroxybutyric acid Carboxylic acid derivatives 0.98*
rhizosphere soil was closely correlated with the microflora 4-Hydroxy benzoic acid Phenolic acid 1.00**
using carbohydrate and phenolic acid as carbon sources. * Significant at P < 0.05. ** Significant at P < 0.01.

Table 2 Correlation analysis between autotoxic allelopathic potential of tobacco rhizospheric soil and AWCD values of
microflora in continuous cropping system
Autotoxic potential AWCD value of microflora cultured in single-carbon source
Carbon source
Tobacco Lettuce Carbohydrate Phenolic acid Fatty acid Carboxylic acid Amide Amino acid
Carbohydrate 0.99** 0.98* 1.00**
Phenolic acid 0.98* 0.99* 0.94 1.00**
Fatty acid 0.94 0.96* 0.89 0.99** 1.00**
Carboxylic acid 0.57 0.51 0.66 0.37 0.25 1.00**
Amide 0.24 0.18 0.36 0.03 0.11 0.94 1.00**
Amino acid 0.64 0.68 0.55 0.79 0.87 0.27 0.59 1.00**
Data are the averages of treatments T1, T2, and T3. * Significant at P < 0.05. ** Significant at P < 0.01.

3 Discussion tobacco rhizopheric soil, and thus affect autotoxicity of the

Continuous monoculture results in serious decline of
biomass and quality of tobacco and enhances occurrence of
disease in production [1]. It is found that fertilization
management can alleviate this continuous cropping obstacle
to some extent, such as proper quantities of fertilizers and
application regime [8–10]. This effect was confirmed in this
study, in which farmyard manure and commercial organic
fertilizer showed positive effects on reducing autotoxic
potential of rhizospheric soil under tobacco continuous
cropping system, particularly, the farmyard manure.
Fertilization is able to change the microflora populations in
soil [8–10]. Microorganism is a component of soil ecology and
plays an important role in nutrient transformation and
decomposition. The structure of soil microbial community
refers to the quantity and proportion of main microflora in soil.
Changes in the community structure directly affect the
functions of rhizospheric microorganisms; as a result, plant
growth is regulated due to variation of soil substance and
energy conversion. Therefore, the effect of fertilizer on
microbial function diversity is a research focus for eliminating
autotoxicity in soil of continuous cropping tobacco. We found
that the soil microflora was increased significantly in number
after fertilization, but its function diversity varied with
 YANG Yu-Hong et al. / Acta Agronomica Sinica, 2011, 37(1): 105–111
fertilizer types. Based on AWCD values of the single-carbon
species, the functional diversity of microflora can be
explained with 2 principal components, and 3 carbon sources
(carbohydrate, fatty acid, and phenolic acid) contributed
mainly to this separation. According to correlation analysis,
the AWCD values of microflora using carbohydrates and
phenolic acid as carbon sources were correlated with the
autotoxic effect of tobacco in the continuous cropping soil.
This finding is helpful to understand the crucial factors in the
complex soil microecosystem of continuous cropping tobacco
that may influence the microbial functional diversity. We
concluded that the microorganisms using carbohydrates and
phenolic acids as carbon sources play a key role in the
autotoxic effect of tobacco in continuous cropping soil.
Plant continuous cropping may cause variations of
chemical substances in rhizospheric soils. For example, the
continuous cropping of poplar (Populus deltoides Bart. ex.
Marsh.) increased the phenolic acid accumulation and reduced
the ammonifier in soils [15]; in rhizosphere of continuous
cropping soybean (Glycine max L. Merr.), phenolic acids were
observed in accumulation [16]. In this study, we found that the
rhizospheric microflora using carbohydrate and phenolic acid
were closely correlated with the autotoxic effect of tobacco.
Therefore, we speculated that the accumulations of phenolic
acid and carbohydrate in rhizospheric soil might be a reason
for the continuous cropping obstacle in tobacco. However, the
phenolic acid-dependent microorganism can be stimulated
when farmyard manure is applied, resulting in the
consumption of phenolic acid in soil and alleviation of
autotoxic effect ultimately. Comparatively, traditional
compound fertilizer was favorable for microflora feeding on
amino acid, fatty acid, carboxylic acid, and amine as carbon
sources, but inhibited the growth of microflora using phenolic
acid and carbohydrate (Table 2). The result of this study
showed that farmyard manure was the best type of fertilizer in
the continuous cropping system of tobacco, whereas, the
traditional compound fertilizer was the worst.
In the complex rhizospheric microecosytem of tobacco, a
variety of microorganisms are involved. We need to know
what species could use carbohydrate and phenolic acid as
carbon sources and their interactions. Besides, the mechanism
of signal conduction may explain the stimulation and
inhabitation of favorable microfloar in rhizosphereic soil of
tobacco. These issues should be disclosed in future studies.
4 Conclusions
Fertilizer management could optimize the structure of
microbial community in rhizospheric soil of tobacco and
mitigate the continuous cropping obstacle. Farmyard manure
was the most suitable fertilizer to alleviate such obstacle,
followed by commercial organic fertilizer, and traditional
compound fertilizer was the worst. Farmyard manure was
favorable to the growth of the microflora feeding on
carbohydrate and phenolic acid as carbon sources, which
might be the key factors for mitigating the continuous
cropping obstacle of tobacco.
Acknowledgment
The study was supported by the projects of Yunnan
Provincial Tobacco Monopoly Bureau, Kunming, China.
References
[1] Jin Y, Yang Y H, Duan Y Q, Kong G H. Effect of rotational
cropping and continuous cropping on yield and quality of
flue-cured tobacco. Southwest China J Agric Sci, 2004,
17(suppl-1): 267–271 (in Chinese with English abstract)
[2] Freddy A. Common bean response to tillage intensity and weed
control strategies. Crop Syst, 2001, 93: 556–563
[3] Yu J Q. Autotoxic potential in vegetables crops. In Ailelopathy
Update-basic and Applied Aspects. Ed. S. & Narwad. 1999. pp
149–162
[4] Zhang X L, Pang Z G, Zhou X F, Ni W Z. Continuous cropping
obstacle and autotoxicity. Chin J Soil Sci, 2007, 38: 267–271
(in Chinese with English abstract)
[5] Wu F Z, Zhao F Y, Gu S Y. Effect of the continuous
cultivating cucumber on the bio-chemical properties of soil in
the plastic greenhouse. Syst Sci Comprehensive Studies Agric,
2002, 18: 20–22 (in Chinese with English abstract)
[6] Pan M Y, Zhang Y Z, Xiao N Q, Tan Z J. Effect of continuous
cropping tobacco on the microbes and enzyme activities in dry
land soil. World Sci-Tech R&D, 2008, 30: 295–297 (in Chinese
with English abstract)
[7] Chen D M, Ke W H, Chen L L, Huang J W, Wu W X, Chen T,
Zhang Z Y, Lin W X. Diversity of bacterial community in
rhizosphere soils under effects of continuously planting burley
tobacco. Chin J Appl Ecol, 2010, 21: 1751–1758 (in Chinese
with English abstract)
[8] Xue X P, Zhang Q, Chen Y, Wang M S, Pan W J, Li J X.
Effects of fertilization and microorganism on growth of
continuous cropping tobacco seedling. Acta Agric Jiangxi,
2008, 20(9): 1–4 (in Chinese with English abstract)
[9] Wang C, Wu F, Liu X L, Liu B. Tobacco rhizosphere
microorganism in different fertility of soil. Chin Tob Sci, 2005,
(2): 12–14 (in Chinese with English abstract)
[10] Guo H H, Yi J H, Jia Z H, Sun Z J, Li H W. Effect of
fertilizaion on tobacco growth and microbe quantity in
rhizosphere soil. Tob Sci Technol, 2004, (6): 40–42 (in Chinese
with English abstract)
[11] Wu J F, Lin X G. Effect of soil microbes on plant growth. Soils,
2001, (1): l8–21 (in Chinese with English abstract)
[12] Guo H X, Liu W Q, Jiang Z S. Effect of cake seed sesame
fertilizer on soil microbe in root system of flue-cured tobacco.
J Henan Agric Univ, 2002, (4): 344–347 (in Chinese with
English abstract)
[13] Xia Z Y, Li Y H, Yang S J. Effects of microbial fertilizer on
flue-cured tobacco on field. Chin Tob Sci, 2002, (3): 28–30 (in
 YANG Yu-Hong et al. / Acta Agronomica Sinica, 2011, 37(1): 105–111

Chinese with English abstract)
[14] Lin R Y, Rong H, Zhou J J, Yu C P, Ye C Y, Chen L S, Lin W
X. Impact of rice seedling allelopathy on rhizospheric microbial
populations and their functional diversities. Acta Ecol Sin,
2007, 27: 3644–3654 (in Chinese with English abstract)
[15] Tan X M, Wang H T, Kong L G, Wang Y P. Accumulation of
phenolic acids in soil of a continuous cropping Poplar
plantation and their effects on soil microbes. J Shandong Univ
(Nat Sci), 2008, 43(1): 14–19 (in Chinese with English
abstract)
[16] Zhang S X, Gao Z Q, Liu H L. Continuous cropping obstacle
and rhizospheric microecology: III. Soil phenolic acids and
their biological effect. Chin J Appl Ecol, 2000, 11(5): 102–105
(in Chinese with English abstract)