• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Soil microbial characteristics and yield response to partial substitution of chemical fertilizer with organic amendments in greenhouse vegetable production

    2018-06-06 09:13:17RONGQinleiLlRuonanHUANGShaowenTANGJiweiZHANGYancaiWANGLiying
    Journal of Integrative Agriculture 2018年6期

    RONG Qin-lei, Ll Ruo-nan, HUANG Shao-wen, TANG Ji-wei, ZHANG Yan-cai, WANG Li-ying

    1 Key Laboratory of Plant Nutrition and Fertilizer, Ministry of Agriculture/Institute of Agricultural Resources and Regional Planning,Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China

    2 Institute of Agricultural Resources and Environment, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050051,P.R.China

    1. lntroduction

    In China, greenhouse vegetable production has undergone rapid development from less than 7 000 hectares in 1980 to almost 3.7 million hectares in 2015 (Huanget al.2016).The amount of inorganic nitrogen (N) fertilizer used on vegetable fields is greater than that used on cereal crops(Juet al.2006; Minet al.2011). This increased application of inorganic N can cause environmental pollution and soil degradation (Shenet al.2010; Rezaei Rashtiet al.2015).To avoid these problems, applying chemical fertilizers that are partially substituted with organic amendments is an economical and effective practice (Syswerdaet al.2012;Aparnaet al.2014). However, as an intensive and unique form of agriculture, greenhouse vegetable production has been characterized by a high cropping index and high agricultural inputs (Liet al.2001; Zhuet al.2011; Yanget al.2014), which is different from open fields in the same region. As a result, nutrient cycling induced by microbial activities undergoes large, relative changes in greenhouse vegetable production (Linet al.2004; Qinet al.2016; Yaoet al.2016). Thus, a better understanding of how partial substitution with organic amendments affects activity and microbial community composition of soil is needed in order to determine the implications for managing greenhouse agroecosystems.

    Soil microbes make important contributions to biologicallyand biochemically-mediated processes in soil, e.g., the decomposition of soil organic matter and nutrient retention(Nannipieriet al.2003; K?hl and van der Heijden 2016).Microbial parameters such as microbial biomass, enzyme activities, and activity and composition of the microbial community are sensitive and significant indicators for assessing changes in soil quality (Sharmaet al.2015).Soil microbial biomass, as a living part of soil organic matter, is an early indicator of soil management changes.Several long-term experiments have found that organic fertilization increases microbial biomass because the organic amendment delivers large amounts of external carbon (C) to soil (Kaschuket al.2010; Zhanget al.2015).However, the response of the microbial biomass to organic C input depends upon the application rate and the chemical composition of the organic amendments used (Kallenbach and Grandy 2011). Therefore, more information is needed on the effects of different inputs of organic amendments as substitutes on soil microbial biomass in greenhouse vegetable production.

    Phospholipid fatty acid (PLFA) analysis, as an established technique, has been widely used to evaluate the influence of management practices or environmental factors on changes in the composition of microbial communities (Zelles 1999). The relative abundances of Gram-negative bacteria,saprophytic fungi, and actinomycetes were related to soil organic C (SOC) transformation or turnover in the forest soil of the Baotianman Nature Reserve (Youet al.2014).The application of chemical N fertilizer increased the total microbial biomass and fungal abundance but decreased the bacterial abundance in a fluvo-aquic soil in a wheat(Triticum aestivumL.)-maize (Zea maysL.) rotation (Aiet al.2012). The influence of management practices such as cover crops, continuous cropping, and compost on soil microbial community composition has attracted attention in greenhouse vegetable production (Maulet al.2014;Willekenset al.2014). Nonetheless, the effect of inorganic fertilizers with different percentages of organic amendments on soil microbial community composition has not been well documented.

    Enzyme activity makes nutrients more available to plants and microorganisms by mineralizing organic C, N, sulphur(S), and phosphorus (P) from soil organic matter (Waringet al.2014). Adding organic material often leads to an overall increase in enzyme activity (Bonanomiet al.2014),but the response depends on changes in soil management and plant cover of soil (Nannipieriet al.2012). Longterm application of horse manure compost to greenhouse vegetable soil enhanced α-galactosidase, β-galactosidase,α-glucosidase, and β-glucosidase activities of soil (Zhanget al.2015). The activities of dehydrogenase, urease, and neutral phosphatase decreased significantly by increasing the rate of N application to a 2-year tomato (Solanum lycopersicumMill.)-cucumber (Cucumis sativusL.) rotation in China’s Yangtze River Delta (Shenet al.2010). At present, most research is focused on enzyme activity related to N and C transformation in greenhouse vegetable production systems. Very few studies have explored the response of soil enzymes involved in the C, N, S, and P biochemical cycles as affected by different percentages of organic amendment substitutes to inorganic fertilizers in greenhouse vegetable production.

    The objective of this work was to examine the effect of using different patterns of organic amendments as partial substitutes for chemical fertilizers on microbial biomass carbon (MBC) and nitrogen (MBN), soil microbial community composition, enzyme activity, and vegetable yields in greenhouse conditions. We hypothesized that changes in microbial properties would depend on different organic amendments (pig manure and straw), and these changes would be related to vegetable yield. To test our hypotheses,a 5-year field experiment was conducted with the following aims: (1) to investigate changes in chemical, biochemical,and microbial properties of soil and vegetable yield and to compare these patterns; and (2) to analyze the relationship between soil microbial properties and vegetable yield.

    2. Materials and methods

    2.1. Site description

    The 5-year experiment was conducted at the Dahe Experimental Station belonging to the Hebei Academy of Agriculture and Forestry Sciences, Hebei Province,China (38°08′N, 114°23′E). The study site has a warm,sub-humid continental monsoon climate with an average annual temperature of 11.5°C and annual precipitation of 540 mm.

    The solar greenhouse employed in the test measured 8 m× 48 m. The original surface soil was used for building the back wall. A crop rotation of winter-spring cucumber(Bomei 11) and autumn-winter tomato (Jinpeng 11) was used in the experiment from October 2009, and the field was left fallow between the growth periods of the two crops. The soil in the study has a clay loam texture and can be classified as calcareous cinnamon soil (FAO classification). The main soil properties (0–20 cm depth) are as follows: soil bulk density,1.35 g cm–3; electrical conductivity, 185.4 mS cm–1; pH, 8.0;organic matter, 9.1 g kg–1; nitrate N, 18.3 mg kg–1; available P, 6.2 mg kg–1; and available K, 98.2 mg kg–1.

    2.2. Experimental design

    A randomized block design was used with 3 replications and 5 different treatments as follows: (1) 4/4CN (CN, N in chemical fertilizer); (2) 3/4CN+1/4MN (MN, N in pig manure);(3) 2/4CN+2/4MN; (4) 2/4CN+1/4MN+1/4SN (SN, N in corn straw); and (5) 2/4CN+2/4SN. The amount of nutrient inputs (N, P2O5, and K2O) for the five treatments was the same. The nutrient inputs were determined based on soil tests and nutrient requirements for target yields (150 t ha–1for cucumber and 90 t ha–1for tomato) (Huanget al.2017).The nutrient inputs of N, P2O5, and K2O used in the winterspring cucumber season were 600, 300, and 525 kg ha–1,respectively, whereas those in the autumn-winter tomato season were 450, 225, and 600 kg ha–1, respectively.

    The chemical fertilizers used in the experiment included urea, calcium superphosphate, and potassium chloride. In order to keep the fertilizer treatments comparable with only small interannual changes every year, fresh commercial pig manure was used, which contained (33.2±4.5)% of water,(1.67±0.13)% of N, (1.30±0.12)% of P2O5, (1.04±0.12)%of K2O, and (142.6±6.8) g kg–1of C. The fresh straw used contained ((13.7±1.6)% of water, (0.75±0.08)%of N, (0.20±0.03)% of P2O5, (1.21±0.22)% of K2O, and(324.1±12.3) g kg–1of C. In the winter-spring cucumber season, 9 t ha–1of fresh commercial pig manure was used in the treatments 3/4CN+1/4MN and 2/4CN+1/4MN+1/4SN,of which 25% of the N came from pig manure; 20 t ha–1of fresh straw was used in the 2/4CN+1/4MN+1/4SN treatment,of which 25% of the N came from straw. In the autumnwinter tomato season, 6.75 t ha–1of fresh commercial pig manure was used in the treatments 3/4CN+1/4MN and 2/4CN+1/4MN+1/4SN, of which 25% of the N came from pig manure; and 15 t ha–1of fresh straw was used in the 2/4CN+1/4MN+1/4SN treatment, of which 25% of the N came from straw. When 50% N was substituted, the consumption of pig manure and straw doubled. Inputs of N,P2O5, and K2O for each organic amendment treatment were calculated by each nutrient concentration in pig manure and corn straw and the organic amendment investments. The amounts of N, P2O5, K2O, and C from chemical fertilizer,manure, and straw for each treatment are given in Tables 1 and 2.

    All the pig manure and 20% N, 100% P, and 40% K of the chemical fertilizer were evenly broadcast onto and mixed into the soil through rotary tillage. The crop straw was cut into short pieces and scattered within the 20–25 cm soil layer and then was covered with soil. In the winter-spring cucumber season, the remaining 80% of the N and 60%of the K was divided equally into 10 parts and top-dressed according to the nutrient requirements of the crop. In the autumn-winter tomato seasons, the rest of the fertilizer (80%N; 60% K) was divided into four parts and top-dressed at the expanding stage of the first to fourth fruit spikes when the diameter of the fruits reached 3–4 cm.

    The experimental plot corresponded to an area of 14.4 m2(2.4 m wide×6 m long). In each plot, 4 rows with 20 plants per row were planted (0.6 m between rows, 0.30 m between plants). Polyvinyl chloride (PVC) plates were embedded into the soil between plots to a depth of 100 cm,which extended above the ground by 5 cm. These were used to prevent lateral and transverse migration of nutrients and water between plots. Furrow irrigation was employed in the experiment. A soil probe (WITU technology Inc.,Shenyang, China) was inserted into the soil to a depth of 20 cm to monitor and control the moisture content at 75–95%of the field capacity.

    2.3. Soil sampling and analyses

    Sampling protocolSoil samples were collected from each plot at 0–20 cm depth during the uprooting stage of the fifth crop cycle (10 July 2014). Fresh soil samples were immediately transported to the laboratory in an ice-box for further processing. Gravel and residual roots were removed,and the soil samples were sieved (2-mm). A portion of each sample was then stored at 4°C to be analyzed for enzyme activities, soil microbial biomass, and dissolved organic C(DOC) and N (DON). Another part was stored at –70°C for microbiological PLFA analysis. The rest of each sample was air-dried for chemical analysis.

    Soil analysesBoth MBC and MBN were determined using the fumigation-extraction technique (Wuet al.1990),while the filtrates were analyzed using a TOC/TN analyzer(Analytik Jena, Multi N/C 3100). Both of the efficiencyconstants required for MBC and MBN calculation,kECandkEN, were taken to be 0.45 (Wuet al.1990; Joergensen 1996). The SOC content was determined by oxidation with potassium dichromate and titration with ferrous ammonium sulfate. The nitrate-N level (NO3–-N) was determined using the dual-wavelength ultraviolet spectrophotometric method(Normanet al.1985). DOC and DON were analyzed as described by Ghaniet al.(2007).

    Table 1 Amounts of nitrogen (N), phosphorus (P2O5), and potassium (K2O) used in each treatment during the winter-spring cucumber season

    Table 2 Amounts of nitrogen (N), phosphorus (P2O5), and potassium (K2O) used in each treatment during the autumn-winter tomato season

    Enzyme activityAll enzyme activities with the exception of urease, phenol oxidase, and peroxidase activities were determined using the microplate fluorometric assay(DeForest 2009; Aiet al.2015). The fluorescence was quantified using a microplate fluorometer (Scientific Fluoroskan Ascent FL, Thermo, USA) with 365 nm excitation and 450 nm emission filters (Saiya-Corket al.2002). The enzyme activities were expressed in nmol h–1g–1. Urease activity was determined using urea as the substrate as described by Kandeler and Gerber (1988) and was expressed as mmol NH4+g–1dry soil h–1.

    PLFA analysisThe composition of the soil microbial community was determinedviaPLFA analysis according to the procedure described by Wuet al.(2009). The abundance of individual PLFAs was indicated by its %mole abundance in each sample. PLFAs were divided into various taxonomic groups based on previously published PLFA biomarker data (Aiet al.2012; Moeskopset al.2012).Specifically, we used i14:0, a15:0, i15:0, 16:0, i16:0, 17:0,a17:0, cy17:0, i17:0, and cy19:0ω8c as bacteria biomarkers;i14:0, a15:0, i15:0, i16:0, a17:0, and i17:0 as Gram-positive bacteria biomarkers; and cy17:0 and cy19:0ω8c as Gramnegative bacteria biomarkers. The unsaturated PLFA 18:1ω9c was used as a fungal biomarker. The fatty acids 16:0 (10Me), 17:0 (10Me), and 18:0 (10Me) were used as markers for actinomycetes.

    2.4. Statistical analysis

    Statistical analysis was carried out using the SAS software package (ver. 9.1). A two-way randomized block ANOVA test was performed to analyze each variable using Fisher’s least significant difference (P=0.05) to compare the treatment means. Pearson correlation analysis was performed to determine the relationship of MBC and MBN, enzyme activity, microbial community composition, and yield (5 different treatments of the fifth year, three replicates each).Principal component analysis (PCA) and redundancy analysis (RDA) with the Monte Carlo permutation test (499 permutations) were performed to determine if soil enzyme activity or community composition was correlated with soil properties, as implemented in Canoco 5.0.

    3. Results

    3.1. Changes in vegetable yield

    The cucumber and tomato yields from 2010 to 2014 are shown in Table 3. The 4/4CN treatment had higher vegetable yield than treatments using organic amendment substitutions in the first three growing seasons. Since the autumn-winter tomato season in 2012,straw treatment (2/4CN+2/4SN) significantly increased tomato yield compared with 4/4CN treatment (except in the autumn-winter tomato season in 2013). For cucumber yield,straw treatment (2/4CN+1/4MN+1/4SN, 2/4CN+2/4SN)induced a slight increase in the fifth growing season in 2012, however, a significant increase was observed in the following winter-spring cucumber seasons in 2013 and 2014.However, there was no significant difference between 2/4CN+1/4MN+1/4SN and 2/4CN+2/4SN.

    3.2. Changes in microbial biomass C and N

    Both MBC and MBN were significantly affected by the organic amendments in the ninth growing season (except 3/4CN+1/4MN) (Fig. 1). The MBC and MBN values in organic-amended soil (i.e., 3/4CN+1/4MN, 2/4CN+2/4MN,2/4CN+1/4MN+1/4SN, and 2/4CN+2/4SN), were in the ranges of 137.0–290.2 and 36.2–57.2 mg kg–1, respectively.These values are much higher than those in 4/4CN treatment, which were in the ranges of 10.5–134.1 and 28.1–102.1%, respectively (Fig. 1).

    Both MBC and MBN also increased with higher amounts of added pig manure or straw (Fig. 1). The MBC and MBN in the 2/4CN+2/4MN treatment were, on average,25.2 and 32.8% higher, respectively, than those in the 3/4CN+1/4MN treatment. Similarly, MBC and MBN in the 2/4CN+2/4SN treatment were, on average, 27.9 and 6.7%higher, respectively, than those in the 2/4CN+1/4MN+1/4SN treatment.

    Table 3 The effects of different partial replacements of inorganic fertilizer (t ha–1) on the yields of winter-spring cucumber and autumn-winter tomato in the greenhouses from 2010 to 2014

    Fig. 1 The effects of different partial organic-amendment replacements on microbial biomass carbon (A) and nitrogen (B) in the soil in the uprooting stage of the 9th growing season (winter-spring cucumber). CN, nitrogen in chemical fertilizer; MN, nitrogen in pig manure; SN, nitrogen in corn straw. Bars indicate standard error, n=3. Different letters indicate significant differences between treatments (at the P< 0.05 level).

    3.3. Changes in soil enzyme activity

    Enzymes involved in C, N, and P cycling were significantly increased in activity in the organic-amended soil. The largest soil enzyme activities were observed in soils amended with straw (Fig. 2). Compared with 4/4CN, the activities of β-glucosidase, β-cellobiosidase, β-xylosidase,and α-glucosidase (involved in the C cycle) were,respectively, increased by 130.9–477.7, 215.1–968.6,81.6–412.4, and 47.0–183.9% in soil amended with straw(2/4CN+1/4MN+1/4SN, 2/4CN+2/4SN). The activity of N-acetyl-glucosaminidase, L-leucine aminopeptidase, and urease (related to the N cycle) were increased by 58.4–545.2, 0.1–37.8, and 38.8–188.5%, respectively. In contrast,the activity of soil phosphomonoesterase (involved in the P cycle) increased by 8.6–162.9% (Fig. 2). Sulfatase activity in the organic-amended soil was significantly decreased,especially when straw was used. The sulfatase activity fell by 2.1–9.3% compared to 4/4CN, and by 1.2–7.4%compared to pig-manure treated soil (Fig. 2).

    Ordination of the fertilization treatmentsviaPCA shows that they are primarily related to the first canonical axis(PC1) (PC1=95.1%). The samples were separated into three distinct groups, each possessing a specific range of soil MBC values (Fig. 3-A). The first group included 4/4CN,3/4CN+1/4MN, and 2/4CN+2/4MN treated soil, i.e., those generally having lower MBC values of 124.0–171.6 mg kg–1(Fig. 3-A). The second group included soil treated with 2/4CN+1/4MN+1/4SN which had an intermediate MBC value of 227.0 mg kg–1. The third group, containing 2/4CN+1/4MN+1/4SN treated soil, had a high MBC value of 290.2 mg kg–1. Indeed, RDA confirmed that soil MBC had a statistically significant effect (F=110,P<0.01) on enzyme activity, and that it accounted for 89.5% of the total enzyme activity variation (Fig. 3-B). In addition, DON was also significantly related to enzyme activity (F=2.3,P=0.03)and accounted for 2.8% of the variation in the total enzyme activity (Fig. 3-B).

    3.4. Changes in abundance and composition of the microbial communities

    Treatments using different organic-amendment substitutes increased the total PLFA content, though to different degrees, with the increases ranging from 7.0 to 66.1%compared with 4/4CN (Fig. 4-A). In particular, the soils treated with 50% N substituted by straw (2/4CN+2/4SN)had significantly higher PLFA content than that in 4/4CN treatment.

    Fig. 2 The response of enzyme activity to different partial organic-amendment replacements in the uprooting stage of the 9th growing season. CN, nitrogen in chemical fertilizer; MN, nitrogen in pig manure; SN, nitrogen in corn straw. Bars indicate standard error, n=3. Different letters indicate significant differences between treatments at P<0.05.

    The relative abundance of bacteria was significantly higher in the 2/4CN+2/4SN-treated soil than that in 4/4CN-treated soil, whereas there were no differences between pig manure and straw-treated soils. Abundance of fungi did not show a clear trend in response to organic-amended soil, except for a small but significant difference between 2/4CN+1/4MN+1/4SN and 4/4CN-treated soil. Compared with the 4/4CN-treated soil, the relative abundances of actinomycetes were remarkably increased in the strawtreated soil and the soils treated with 50% N substituted by pig manure. The ratio of Gram-positive to Gram-negative bacteria and bacteria to actinomycetes was significantly decreased in straw-treated soils (2/4CN+1/4MN+1/4SN and 2/4CN+2/4SN) compared to that in 4/4CN-treated soil.However, there were no significant differences in these ratios between pig manure-treated and 4/4CN-treated soil.

    PCA ordination analysis showed that the fertilization treatments could be separated into two distinct groups,each possessing a specific range of soil MBC values(Fig. 5-A). The first group (4/4CN, 3/4CN+1/4MN, and 2/4CN+2/4MN treated soil) generally had lower MBC,ranging from 124.0 to 171.6 mg kg–1(Fig. 5-A). The second group (2/4CN+1/4MN+1/4SN and 2/4CN+2/4SN treated soil) had higher MBC, ranging from 227.0 to 290.2 mg kg–1. The difference is significantly related to the change in soil MBC (F=14.3,P<0.01) (Fig. 5-B), which accounted for 52.3% of the total variation in the composition of the microbial community, based on the RDA between microbial community and soil property.

    3.5. Correlations between yield and soil microbial properties

    Fig. 3 Results of principal component analysis (PCA) of the enzyme activity in soil subjected to different fertilization treatments(A), and redundancy analysis (RDA) of the correlation between soil parameters and enzyme activity profiles (B). MBC, microbial biomass carbon; nitrate-N, NO3–-N; DON, dissolved organic nitrogen; TOC, total organic carbon; MBN, microbial biomass nitrogen;DOC, dissolved organic carbon. CN, nitrogen in chemical fertilizer; MN, nitrogen in pig manure; SN, nitrogen in corn straw. The data are from the uprooting stage of the ninth growing season (winter-spring cucumber). Red arrows indicate soil parameters that have a strong and significant impact on enzyme activity (P< 0.05). The corresponding proportion of the variation accounted for is shown in the lower right corner.

    Fig. 4 A comparison of the total phospholipid fatty acid (PLFA; A), the relative abundance of bacteria (histograms) and the G+ :G– ratio (dotted line; B), the relative abundance of fungi (C) and the relative abundance of actinomycetes (histograms) and bacteria:actinomycetes ratio (dotted line; D). G+, Gram-positive bacteria; G–, Gram-negative bacteria; CN, nitrogen in chemical fertilizer; MN, nitrogen in pig manure; SN, nitrogen in corn straw. Vertical bars represent the standard error (n=3) and lower case letters indicate significant differences between fertilizer treatments at the P< 0.05 level.

    Fig. 5 Results of principal component analysis (PCA) of the microbial community composition in soils receiving different fertilization treatments (A), and redundancy analysis (RDA) of the correlation between soil parameters and microbial community composition (B).MBC, microbial biomass carbon; nitrate-N, NO3–-N; TOC, total organic carbon; MBN, microbial biomass nitrogen; DON, dissolved organic nitrogen; DOC, dissolved organic carbon. CN, nitrogen in chemical fertilizer; MN, nitrogen in pig manure; SN, nitrogen in corn straw. The data are from the uprooting stage of the ninth growing season (winter-spring cucumber). Red arrows indicate soil parameters that have a strong and significant impact on the microbial community composition (P< 0.05). The corresponding proportion of the variation accounted for is shown in the lower right corner.

    Overall, the trends found in the total PLFA content,composition of the microbial communities (bacteria,actinomycetes, Gram-positive bacteria, Gram-negative bacteria), and soil enzyme activity showed that these properties generally increased with increasing MBC and MBN (Table 4). A correlation analysis revealed that the activities of most of the enzymes (phosphomonoesterase,β-glucosidase, β-cellobiosidase, N-acetylglucosaminidase,β-xylosidase, α-glucosidase, L-leucine aminopeptidase,and urease) were significantly positively correlated with soil MBC and MBN. However, sulfatase was significantly negatively correlated with MBC and MBN. MBC was significantly positively correlated with the total PLFA content, bacteria, actinomycetes, Gram-positive and Gram-negative bacteria. The analysis also showed that there were significantly positive correlations between enzyme activities (phosphomonoesterase, β-glucosidase,β-cellobiosidase, N-acetylglucosaminidase, β-xylosidase,α-glucosidase, L-leucine aminopeptidase, and urease)and the composition of the microbial communities(bacteria, actinomycetes, Gram-positive bacteria, Gramnegative bacteria). Furthermore, MBC, enzyme activity(phosphomonoesterase, L-leucine aminopeptidase, and urease), and composition of the microbial communities (total PLFA content, bacteria, fungi, actinomycetes, and Gramnegative bacteria) were significantly positively correlated with yield (Table 4).

    4. Discussion

    Many studies on grain cropland and vegetable fields have shown that the use of organic amendments, e.g., manure,compost, straw, can help to increase crop yield and improve soil quality (Bowleset al.2014; Agegnehuet al.2016). In this study, however, we found that crops grown in 4/4CN-treated soil produces the best cucumber and tomato yields in the first three growing seasons. This is mainly because the topsoil was removed when the greenhouse was newly built, and the organic amendments release nutrients more slowly compared to chemical fertilizer (Tianet al.1992). In our study, the highest yield was achieved using the strawsubstituted treatments after three consecutive fertilization management seasons. Thus, successive applications of the organic amendments were required to effectively improve cucumber and tomato yield and soil quality in the new greenhouse. Productivity in agricultural ecosystems relates to the size and activity of the microbial biomass (Anandet al.2015). Soil MBC was found to be significantly correlated with yield in our study. This result confirms previous findings that there is a close relationship between crop yield and microbial biomass in the soil, under both greenhouse (Chenet al.2000) and open field conditions (Anandet al.2015).

    We also found that cucumber and tomato yields were significantly correlated with soil enzymes associated with N (L-leucine aminopeptidase and urease) and P cycling(phosphomonoesterase). This may be because large amounts of organic C are brought into the soil by the manure and straw and because a portion of the N and P absorbed from the soil by microbes and plants will lead to microbes regulating extracellular enzyme production to acquire the limited nutrients (Allisonet al.2007; Bowleset al.2014).Interestingly, indicator PLFAs for Gram-negative bacteria,fungi, and actinomycetes were also significantly and positively correlated with yield. Daiet al.(2013) previously reported that peanut (Arachis hypogaeaLinn.) growth and yield are promoted by an increase in Gram-negative bacteria. A higher proportion of Gram-negative bacteria in the soil is usually interpreted as a shift from oligotrophic to more copiotrophic conditions(Borgaet al.1994; Saetre and B??th 2000).Additionally, actinobacteria can produce a wide variety of antimicrobial metabolites (Basilioet al.2003), which are beneficial to crop growth.These results indicate that manipulating these MBC-mediated microbial properties may be an important method of improving the yield in the greenhouse-vegetable field. Improved soil biological function has been revealed by noting general trends pointing to positive effects on soil enzyme activities and soil respiration in vegetables grown in plastic tunnels (Bonanomiet al.2014). This suggests that soil MBC is a good indicator to assess the change in biological activity of greenhouse-vegetable soil caused by alternatives to inorganic fertilizers.

    Several studies have shown that the composition of the soil microbial community is changed by organic amendments and that these changes are related to the soil C content(Lazcanoet al.2013; Willekenset al.2014).Interestingly, we found that the composition of the microbial community was more strongly affected by straw than pig manure. This may be because the input of new organic matter significantly stimulated the growth of microorganisms in the soil (Shiet al.2015), and because straw provided more organic C than the pig manure(Table 5). It has also been reported that straw may improve the physical properties of the soil by providing nutrients to directly promote microbial growth and enhance the SOC pool (Luet al.2015).It is generally recognized that soil MBC is used to indicate the size of the microbial community(Bastidaet al.2008). However, enhancement of the soil microbial biomass usually occurs through specific groups of microbial communities(e.g., bacteria, actinomycetes) (Donget al.2014). The results acquired in this study show that the relative abundances of bacteria, Grampositive bacteria, Gram-negative bacteria, and actinomycetes were significantly and positively correlated with soil MBC. We also found that the soil microbial community composition (except for fungi) is significantly correlated with all of the soil enzyme activities involved in the C, N,P, and S cycles, indicating a close relationship between microbial community composition and soil enzyme activity (Allisonet al.2007; Burnset al.2013).

    Table 5 Total organic carbon (TOC), dissolved organic carbon (DOC) and nitrogen (DON), and soil nitrate-nitrogen for different partial organic-amendment replacements in the uprooting stage of the 9th growing season (winter-spring cucumber)

    Table 5 Total organic carbon (TOC), dissolved organic carbon (DOC) and nitrogen (DON), and soil nitrate-nitrogen for different partial organic-amendment replacements in the uprooting stage of the 9th growing season (winter-spring cucumber)

    1) CN, nitrogen in chemical fertilizer; MN, nitrogen in pig manure; SN, nitrogen in corn straw.Data are the mean±SE, n=3. Different letters indicate significant differences among treatments at the P<0.05 level.

    –-N (mg kg–1)4/4CN 7.57±0.44 d 50.2±2.1 c 24.5±0.4 c 68.1±10.1 c 3/4CN+1/4MN 9.55±0.10 c 58.3±6.8 c 29.1±0.4 b 81.8±13.2 bc 2/4CN+2/4MN 12.86±0.30 b 68.7±9.3 bc 32.8±1.4 a 111.3±1.1 a 2/4CN+1/4MN+1/4SN 13.30±0.42 b 90.9±5.2 b 34.0±1.5 a 100.9±21.0 ab 2/4CN+2/4SN 14.43±0.52 a 123.4±9.1 a 34.7±2.9 a 90.8±17.6 ab Fertilizer treatments1) TOC (g kg–1) DOC (mg kg–1) DON (mg kg–1) NO3

    5. Conclusion

    This study demonstrates the changes occurring in soil microbial biomass, enzyme activity, microbial community composition, and cucumber and tomato yield as a result of organic amendment substitution in greenhouse production.The development of indicators of soil ecological functions in the greenhouse vegetable soil may help farmers to evaluate and discover an optimal fertilization management strategy to improve soil quality and increase vegetable yield. We have further presented evidence that MBC-induced changes in soil enzyme activity and microbial community composition might be an important mechanism by which vegetable yield may be improved. In particular, straw-substituted treatments can support high and sustainable yields in greenhousevegetable production systems. In summary, considering all of the effects of different proportions of manure and/or straw substitution on microbial characteristics and vegetable yield(and practical feasibility), combined application of chemical fertilizer, manure, and straw (that is, the combination 2/4CN+1/4MN+1/4SN) appears to be a superior fertilization pattern to use for high, sustainable yields in greenhousevegetable production systems.

    Acknowledgements

    This research was supported by the National Key Research and Development Program of China (2016YFD0201001),the earmarked fund for China Agriculture Research System(CARS-23-B02), and the Key Research and Development Program of Shandong Province, China (2017CXGC0206).

    Agegnehu G, Bass A M, Nelson P N, Bird M I. 2016. Benefits of biochar, compost and biochar-compost for soil quality,maize yield and greenhouse gas emissions in a tropical agricultural soil.Science of the Total Environment,543,295–306.

    Ai C, Liang G, Sun J, He P, Tang S, Yang S, Zhou W, Wang X.2015. The alleviation of acid soil stress in rice by inorganic or organic ameliorants is associated with changes in soil enzyme activity and microbial community composition.Biology and Fertility of Soils,51, 465–477.

    Ai C, Liang G, Sun J, Wang X, Zhou W. 2012. Responses of extracellular enzyme activities and microbial community in both the rhizosphere and bulk soil to long-term fertilization practices in a fluvo-aquic soil.Geoderma,173, 330–338.

    Allison V, Condron L, Peltzer D, Richardson S, Turner B. 2007.Changes in enzyme activities and soil microbial community composition along carbon and nutrient gradients at the Franz Josef chronosequence, New Zealand.Soil Biology and Biochemistry,39, 1770–1781.

    Anand K G V, Kubavat D, Trivedi K, Agarwal P K, Wheeler C,Ghosh A. 2015. Long-term application ofJatrophapress cake promotes seed yield by enhanced soil organic carbon accumulation, microbial biomass and enzymatic activities in soils of semi-arid tropical wastelands.European Journal of Soil Biology,69, 57–65.

    Aparna K, Pasha M A, Rao D L N, Krishnaraj P U. 2014.Organic amendments as ecosystem engineers: Microbial,biochemical and genomic evidence of soil health improvement in a tropical arid zone field site.Ecological Engineering,71, 268–277.

    Basilio A, Gonzalez I, Vicente M, Gorrochategui J, Cabello A,Gonzalez A, Genilloud O. 2003. Patterns of antimicrobial activities from soil actinomycetes isolated under different conditions of pH and salinity.Journal of Applied Microbiology,95, 814–823.

    Bastida F, Kandeler E, Moreno J, Ros M, García C, Hernández T. 2008. Application of fresh and composted organic wastes modifies structure, size and activity of soil microbial community under semiarid climate.Applied Soil Ecology,40, 318–329.

    Bonanomi G, D’Ascoli R, Scotti R, Gaglione S A, Caceres M G, Sultana S, Scelza R, Rao M A, Zoina A. 2014. Soil quality recovery and crop yield enhancement by combined application of compost and wood to vegetables grown under plastic tunnels.Agriculture,Ecosystems & Environment,192, 1–7.

    Borga P, Nilsson M, Tunlid A. 1994. Bacterial communities in peat in relation to botanical composition as revealed by phospholipid fatty acid analysis.Soil Biology and Biochemistry,26, 841–848.

    Bowles T M, Acosta-Martínez V, Calderón F, Jackson L E.2014. Soil enzyme activities, microbial communities, and carbon and nitrogen availability in organic agroecosystems across an intensively-managed agricultural landscape.Soil Biology and Biochemistry,68, 252–262.

    Burns R G, DeForest J L, Marxsen J, Sinsabaugh R L,Stromberger M E, Wallenstein M D, Weintraub M N,Zoppini A. 2013. Soil enzymes in a changing environment:Current knowledge and future directions.Soil Biology and Biochemistry,58, 216–234.

    Chen G, He Z, Huang C. 2000. Microbial biomass phosphorus and its significance in predicting phosphorus availability in red soils.Communications in Soil Science & Plant Analysis,31, 655–667.

    Dai C C, Chen Y, Wang X X, Li P D. 2013. Effects of intercropping of peanut with the medicinal plantAtractylodes lanceaon soil microecology and peanut yield in subtropical China.Agroforestry Systems,87, 417–426.

    DeForest J L. 2009. The influence of time, storage temperature,and substrate age on potential soil enzyme activity in acidic forest soils using MUB-linked substrates and L-DOPA.Soil Biology and Biochemistry,41, 1180–1186.

    Dong W, Zhang X, Dai X, Fu X, Yang F, Liu X, Sun X, Wen X,Schaeffer S. 2014. Changes in soil microbial community composition in response to fertilization of paddy soils in subtropical China.Applied Soil Ecology,84, 140–147.

    Ghani A, Dexter M, Carran R, Theobald P. 2007. Dissolved organic nitrogen and carbon in pastoral soils: The New Zealand experience.European Journal of Soil Science,58, 832–843.

    Huang S, Tang J, Li C. 2016. Status of heavy metals in vegetable soils under different patterns of land use.Plant Nutrition and Fertilizer Science,3, 707–718. (in Chinese)

    Huang S, Tang J, Zhang H, Yuan S, Wang Y. 2017. Drip fertigation technology of greenhouse cucumber based on management strategy at different growth stages.China Fruit& Vegetable,37, 82–84. (in Chinese)

    Joergensen R G. 1996. The fumigation-extraction method to estimate soil microbial biomass: Calibration of thekECvalue.Soil Biology and Biochemistry,28, 25–31.

    Ju X T, Kou C L, Zhang F S, Christie P. 2006. Nitrogen balance and groundwater nitrate contamination: Comparison among three intensive cropping systems on the North China Plain.Environmental Pollution,143, 117–125.

    Kallenbach C, Grandy A S. 2011. Controls over soil microbial biomass responses to carbon amendments in agricultural systems: A meta-analysis.Agriculture,Ecosystems &Environment,144, 241–252.

    Kandeler E, Gerber H. 1988. Short-term assay of soil urease activity using colorimetric determination of ammonium.Biology and Fertility of Soils,6, 68–72.

    Kaschuk G, Alberton O, Hungria M. 2010. Three decades of soil microbial biomass studies in Brazilian ecosystems: Lessons learned about soil quality and indications for improving sustainability.Soil Biology and Biochemistry,42, 1–13.

    K?hl L, van der Heijden M G A. 2016. Arbuscular mycorrhizal fungal species differ in their effect on nutrient leaching.Soil Biology and Biochemistry,94, 191–199.

    Lazcano C, Gómez-Brandón M, Revilla P, Domínguez J. 2013.Short-term effects of organic and inorganic fertilizers on soil microbial community structure and function.Biology and Fertility of Soils,49, 723–733.

    Li W, Zhang M, Zee S V D. 2001. Salt contents in soils under plastic greenhouse gardening in China.Pedosphere,11,359–367.

    Lin X G, Yin R, Zhang H Y, Huang J F, Chen R R, Cao Z H.2004. Changes of soil microbiological properties caused by land use changing from rice-wheat rotation to vegetable cultivation.Environmental Geochemistry and Health,26,119–128.

    Lu P, Lin Y, Yang Z, Xu Y, Tan F, Jia X, Wang M, Xu D, Wang X. 2015. Effects of application of corn straw on soil microbial community structure during the maize growing season.Journal of Basic Microbiology,55, 22–32.

    Maul J E, Buyer J S, Lehman R M, Culman S, Blackwood C B, Roberts D P, Zasada I A, Teasdale J R. 2014. Microbial community structure and abundance in the rhizosphere and bulk soil of a tomato cropping system that includes cover crops.Applied Soil Ecology,77, 42–50.

    Min J, Zhao X, Shi W M, Xing G X, Zhu Z L. 2011. Nitrogen balance and loss in a greenhouse vegetable system in southeastern China.Pedosphere,21, 464–472.

    Moeskops B, Buchan D, Sukristiyonubowo De Neve S, De Gusseme B, Widowati L R, Setyorini D, Sleutel S. 2012. Soil quality indicators for intensive vegetable production systems in Java, Indonesia.Ecological Indicators,18, 218–226.

    Nannipieri P, Ascher J, Ceccherini M, Landi L, Pietramellara G, Renella G. 2003. Microbial diversity and soil functions.European Journal of Soil Science,54, 655–670.

    Nannipieri P, Giagnoni L, Renella G, Puglisi E, Ceccanti B,Masciandaro G, Fornasier F, Moscatelli M C, Marinari S. 2012. Soil enzymology: Classical and molecular approaches.Biology and Fertility of Soils,48, 743–762.

    Norman R J, Edberg J C, Stucki J W. 1985. Determination of nitrate in soil extracts by dual-wavelength ultraviolet spectrophotometry.Soil Science Society of America Journal,49, 1182–1185.

    Qin H L, Zhang Z X, Lu J, Zhu Y J, Webster R, Liu X L, Yuan H Z, Hou H J, Chen C L, Wei W X. 2016. Change from paddy rice to vegetable growing changes nitrogen-cycling microbial communities and their variation with depth in the soil.European Journal of Soil Science,67, 650–658.

    Rezaei Rashti M, Wang W, Moody P, Chen C, Ghadiri H. 2015.Fertiliser-induced nitrous oxide emissions from vegetable production in the world and the regulating factors: A review.Atmospheric Environment,112, 225–233.

    Saetre P, B??th E. 2000. Spatial variation and patterns of soil microbial community structure in a mixed spruce–birch stand.Soil Biology and Biochemistry,32, 909–917.

    Saiya-Cork K, Sinsabaugh R, Zak D. 2002. The effects of long term nitrogen deposition on extracellular enzyme activity in anAcer saccharumforest soil.Soil Biology and Biochemistry,34, 1309–1315.

    Sharma S D, Kumar P, Bhardwaj S K, Chandel A. 2015.Agronomic performance, nutrient cycling and microbial biomass in soil as affected by pomegranate based multiple crop sequencing.Scientia Horticulturae,197, 504–515.

    Shen W, Lin X, Shi W, Min J, Gao N, Zhang H, Yin R, He X. 2010. Higher rates of nitrogen fertilization decrease soil enzyme activities, microbial functional diversity and nitrification capacity in a Chinese polytunnel greenhouse vegetable land.Plant and Soil,337, 137–150.

    Shi P, Wang S, Jia S, Gao Q. 2015. Effect of 25-year fertilization on soil microbial biomass and community structure in a continuous corn cropping system.Archives of Agronomy and Soil Science,61, 1303–1317.

    Syswerda S P, Basso B, Hamilton S K, Tausig J B, Robertson G P. 2012. Long-term nitrate loss along an agricultural intensity gradient in the Upper Midwest USA.Agriculture,Ecosystems & Environment,149, 10–19.

    Tian G, Kang B, Brussaard L. 1992. Biological effects of plant residues with contrasting chemical compositions under humid tropical conditions-decomposition and nutrient release.Soil Biology and Biochemistry,24, 1051–1060.

    Waring B G, Weintraub S R, Sinsabaugh R L. 2014.Ecoenzymatic stoichiometry of microbial nutrient acquisition in tropical soils.Biogeochemistry,117, 101–113.

    Willekens K, Vandecasteele B, Buchan D, De Neve S. 2014.Soil quality is positively affected by reduced tillage and compost in an intensive vegetable cropping system.Applied Soil Ecology,82, 61–71.

    Wu J, Joergensen R, Pommerening B, Chaussod R, Brookes P. 1990. Measurement of soil microbial biomass C by fumigation-extraction - an automated procedure.Soil Biology and Biochemistry,22, 1167–1169.

    Wu Y, Ding N, Wang G, Xu J, Wu J, Brookes P C. 2009.Effects of different soil weights, storage times and extraction methods on soil phospholipid fatty acid analyses.Geoderma,150, 171–178.

    Yang L, Huang B, Hu W, Chen Y, Mao M, Yao L. 2014. The impact of greenhouse vegetable farming duration and soil types on phytoavailability of heavy metals and their health risk in eastern China.Chemosphere,103, 121–130.

    Yao Z, Xing J, Gu H, Wang H, Wu J, Xu J, Brookes P C. 2016.Development of microbial community structure in vegetablegrowing soils from open-field to plastic-greenhouse cultivation based on the PLFA analysis.Journal of Soils and Sediments,16, 2041–2049.

    You Y, Wang J, Huang X, Tang Z, Liu S, Sun O J. 2014. Relating microbial community structure to functioning in forest soil organic carbon transformation and turnover.Ecology &Evolution,4, 633–647.

    Zelles L. 1999. Fatty acid patterns of phospholipids and lipopolysaccharides in the characterisation of microbial communities in soil: A review.Biology and Fertility of Soils,29, 111–129.

    Zhang L, Chen W, Burger M, Yang L, Gong P, Wu Z. 2015.Changes in soil carbon and enzyme activity as a result of different long-term fertilization regimes in a greenhouse field.PLoS ONE,10, e0118371.

    Zhu T, Zhang J, Cai Z. 2011. The contribution of nitrogen transformation processes to total N2O emissions from soils used for intensive vegetable cultivation.Plant and Soil,343, 313–327.

    亚洲午夜精品一区,二区,三区| 国产精品香港三级国产av潘金莲| 91成年电影在线观看| 亚洲精品久久国产高清桃花| 成熟少妇高潮喷水视频| 国产精品乱码一区二三区的特点| 午夜免费成人在线视频| 舔av片在线| 久9热在线精品视频| 欧美另类亚洲清纯唯美| 国产精品亚洲美女久久久| 国产99白浆流出| 丰满的人妻完整版| 亚洲国产精品成人综合色| 国产熟女午夜一区二区三区| 中文在线观看免费www的网站 | 这个男人来自地球电影免费观看| 国产高清videossex| 又黄又爽又免费观看的视频| 国产精品香港三级国产av潘金莲| 亚洲国产欧美一区二区综合| 欧美日韩亚洲国产一区二区在线观看| 天天一区二区日本电影三级| 亚洲成人精品中文字幕电影| 欧美日韩乱码在线| 人妻久久中文字幕网| 国产精品亚洲美女久久久| 日本五十路高清| 欧美一区二区国产精品久久精品 | 欧美极品一区二区三区四区| 人成视频在线观看免费观看| 国产精品爽爽va在线观看网站| 亚洲中文av在线| 美女高潮喷水抽搐中文字幕| 操出白浆在线播放| 18禁黄网站禁片午夜丰满| 国产精华一区二区三区| 美女 人体艺术 gogo| 最近在线观看免费完整版| 成人av在线播放网站| 99riav亚洲国产免费| 91国产中文字幕| 久99久视频精品免费| 桃色一区二区三区在线观看| 可以免费在线观看a视频的电影网站| 国产亚洲精品综合一区在线观看 | 免费在线观看亚洲国产| 搞女人的毛片| 99久久精品热视频| 免费看十八禁软件| 50天的宝宝边吃奶边哭怎么回事| 久久久久久九九精品二区国产 | 亚洲专区字幕在线| 久久久国产欧美日韩av| 国产亚洲精品久久久久5区| 国产精品亚洲av一区麻豆| 黑人操中国人逼视频| 国产精品影院久久| 亚洲精品中文字幕一二三四区| 91麻豆av在线| 国产精品亚洲一级av第二区| 亚洲熟妇熟女久久| 桃色一区二区三区在线观看| 成人永久免费在线观看视频| 亚洲成人中文字幕在线播放| 狠狠狠狠99中文字幕| www国产在线视频色| 特级一级黄色大片| 国产精品香港三级国产av潘金莲| 男插女下体视频免费在线播放| 午夜久久久久精精品| 中文亚洲av片在线观看爽| 久久天堂一区二区三区四区| 99在线视频只有这里精品首页| 日韩大尺度精品在线看网址| 久久久久亚洲av毛片大全| a级毛片a级免费在线| 97超级碰碰碰精品色视频在线观看| 91九色精品人成在线观看| 好男人电影高清在线观看| 久久香蕉国产精品| 久久久精品国产亚洲av高清涩受| 中文资源天堂在线| 丁香欧美五月| 亚洲一码二码三码区别大吗| 成年女人毛片免费观看观看9| 熟妇人妻久久中文字幕3abv| 欧美午夜高清在线| 热99re8久久精品国产| netflix在线观看网站| 国产亚洲精品第一综合不卡| 欧美在线一区亚洲| 国内精品久久久久精免费| 国产精品综合久久久久久久免费| 久久中文字幕一级| 亚洲国产中文字幕在线视频| 在线观看www视频免费| 99在线人妻在线中文字幕| 99在线视频只有这里精品首页| 天堂av国产一区二区熟女人妻 | 1024手机看黄色片| 一级毛片女人18水好多| 最好的美女福利视频网| 免费观看人在逋| 99久久综合精品五月天人人| 在线观看66精品国产| 99热只有精品国产| 精品一区二区三区四区五区乱码| 国产亚洲精品av在线| 亚洲国产精品999在线| 狠狠狠狠99中文字幕| svipshipincom国产片| 女同久久另类99精品国产91| 国产精品日韩av在线免费观看| 无人区码免费观看不卡| 不卡av一区二区三区| 国产不卡一卡二| 久久久久国内视频| 看片在线看免费视频| av天堂在线播放| 国产亚洲精品久久久久5区| 精品久久久久久成人av| 一进一出抽搐gif免费好疼| 久久精品综合一区二区三区| 可以免费在线观看a视频的电影网站| 老司机在亚洲福利影院| 久久中文看片网| 精品欧美国产一区二区三| 一级毛片精品| 无遮挡黄片免费观看| 大型av网站在线播放| 91大片在线观看| 黄色 视频免费看| 琪琪午夜伦伦电影理论片6080| 欧美3d第一页| 午夜激情av网站| 亚洲成人国产一区在线观看| av片东京热男人的天堂| 亚洲熟女毛片儿| 麻豆国产97在线/欧美 | 亚洲乱码一区二区免费版| 狠狠狠狠99中文字幕| 一本精品99久久精品77| 无遮挡黄片免费观看| a级毛片在线看网站| 久久久精品大字幕| 精品久久久久久,| 国产精品久久视频播放| 在线观看舔阴道视频| 在线观看一区二区三区| 可以在线观看的亚洲视频| 熟妇人妻久久中文字幕3abv| 99热只有精品国产| 久久草成人影院| 欧美成人午夜精品| 国产精品av久久久久免费| 午夜成年电影在线免费观看| 神马国产精品三级电影在线观看 | 色哟哟哟哟哟哟| 亚洲人成77777在线视频| 色综合欧美亚洲国产小说| 在线观看免费视频日本深夜| 成在线人永久免费视频| 观看免费一级毛片| 村上凉子中文字幕在线| 亚洲五月天丁香| 脱女人内裤的视频| 日韩欧美精品v在线| cao死你这个sao货| 精品日产1卡2卡| 日韩国内少妇激情av| 老汉色∧v一级毛片| 久久久精品欧美日韩精品| www.精华液| 国产一级毛片七仙女欲春2| 视频区欧美日本亚洲| www国产在线视频色| 国产亚洲av高清不卡| 色综合站精品国产| 高清在线国产一区| 女人高潮潮喷娇喘18禁视频| 亚洲欧美日韩高清专用| 手机成人av网站| 国产欧美日韩精品亚洲av| 性欧美人与动物交配| 在线十欧美十亚洲十日本专区| 亚洲人成网站在线播放欧美日韩| 非洲黑人性xxxx精品又粗又长| 国产一区二区三区在线臀色熟女| 免费观看人在逋| 国内精品久久久久久久电影| 天堂av国产一区二区熟女人妻 | 午夜两性在线视频| www日本黄色视频网| 老鸭窝网址在线观看| 亚洲欧美激情综合另类| 人妻丰满熟妇av一区二区三区| 久热爱精品视频在线9| 丰满人妻一区二区三区视频av | 日日爽夜夜爽网站| 亚洲精品在线美女| 欧美午夜高清在线| 欧美成人一区二区免费高清观看 | 亚洲av片天天在线观看| 精品人妻1区二区| 日韩欧美免费精品| 久久久久国产一级毛片高清牌| 亚洲午夜理论影院| 久久欧美精品欧美久久欧美| 天天添夜夜摸| 久久这里只有精品19| 蜜桃久久精品国产亚洲av| 亚洲天堂国产精品一区在线| 成人欧美大片| 999精品在线视频| 国产成年人精品一区二区| 18禁美女被吸乳视频| 欧美日韩福利视频一区二区| 人妻丰满熟妇av一区二区三区| 亚洲色图av天堂| 国产av一区二区精品久久| 国模一区二区三区四区视频 | xxxwww97欧美| 欧美乱码精品一区二区三区| 波多野结衣高清无吗| 国产一区二区三区在线臀色熟女| 99久久国产精品久久久| 日日夜夜操网爽| 每晚都被弄得嗷嗷叫到高潮| 亚洲精品美女久久久久99蜜臀| bbb黄色大片| 亚洲欧洲精品一区二区精品久久久| 韩国av一区二区三区四区| 女人高潮潮喷娇喘18禁视频| 听说在线观看完整版免费高清| 欧美色欧美亚洲另类二区| 亚洲午夜理论影院| 欧美日本亚洲视频在线播放| 国产av不卡久久| 久久精品91无色码中文字幕| 亚洲男人的天堂狠狠| 韩国av一区二区三区四区| 最好的美女福利视频网| 男女之事视频高清在线观看| 国产成人啪精品午夜网站| 久久香蕉精品热| 免费av毛片视频| 日韩大码丰满熟妇| 两个人看的免费小视频| 一区二区三区高清视频在线| 91成年电影在线观看| 成人国语在线视频| 国内精品一区二区在线观看| 草草在线视频免费看| 男女视频在线观看网站免费 | 黄色成人免费大全| 我的老师免费观看完整版| av国产免费在线观看| 琪琪午夜伦伦电影理论片6080| 免费在线观看亚洲国产| 亚洲最大成人中文| 久久精品aⅴ一区二区三区四区| 日日干狠狠操夜夜爽| 久久久国产成人精品二区| 亚洲国产日韩欧美精品在线观看 | 久久久久国内视频| 日本在线视频免费播放| 亚洲av五月六月丁香网| 国产精品香港三级国产av潘金莲| 亚洲国产精品成人综合色| 亚洲欧美日韩高清在线视频| 国产99白浆流出| 亚洲国产欧洲综合997久久,| 免费看十八禁软件| 国产精品 欧美亚洲| 少妇熟女aⅴ在线视频| 国产精品影院久久| 露出奶头的视频| 99精品久久久久人妻精品| 国产99久久九九免费精品| 黄色 视频免费看| 亚洲av日韩精品久久久久久密| av片东京热男人的天堂| 午夜福利在线在线| 久久香蕉激情| 久久婷婷成人综合色麻豆| 老鸭窝网址在线观看| 一本大道久久a久久精品| 久久久久久久午夜电影| 久久 成人 亚洲| 日韩欧美三级三区| 日韩欧美国产在线观看| 久久精品夜夜夜夜夜久久蜜豆 | 亚洲欧美精品综合一区二区三区| 99精品欧美一区二区三区四区| 国产三级在线视频| 欧洲精品卡2卡3卡4卡5卡区| 国产99久久九九免费精品| 亚洲精品在线观看二区| 久久精品成人免费网站| 亚洲在线自拍视频| 亚洲真实伦在线观看| 日本 欧美在线| 男男h啪啪无遮挡| 国产精品免费一区二区三区在线| 两人在一起打扑克的视频| 在线观看www视频免费| 日韩欧美免费精品| 欧美一级a爱片免费观看看 | 好男人在线观看高清免费视频| 少妇熟女aⅴ在线视频| 国产三级黄色录像| 国产精品永久免费网站| 此物有八面人人有两片| 日本一二三区视频观看| 亚洲国产欧美网| 国产免费av片在线观看野外av| 国产成人一区二区三区免费视频网站| 亚洲欧美精品综合一区二区三区| 国产精品九九99| 欧美日韩亚洲国产一区二区在线观看| 全区人妻精品视频| 听说在线观看完整版免费高清| 国产精品一区二区免费欧美| 久久婷婷成人综合色麻豆| 亚洲性夜色夜夜综合| 成人国语在线视频| 午夜激情av网站| 国产精品日韩av在线免费观看| 日本一区二区免费在线视频| 中文字幕av在线有码专区| 每晚都被弄得嗷嗷叫到高潮| 长腿黑丝高跟| 日本一二三区视频观看| 精品久久久久久久人妻蜜臀av| 日本黄大片高清| 麻豆国产97在线/欧美 | 老司机深夜福利视频在线观看| 搡老岳熟女国产| 嫁个100分男人电影在线观看| 怎么达到女性高潮| 神马国产精品三级电影在线观看 | 国产99白浆流出| 黄片大片在线免费观看| 亚洲一区二区三区色噜噜| 禁无遮挡网站| 每晚都被弄得嗷嗷叫到高潮| 国产精品美女特级片免费视频播放器 | 看免费av毛片| 久久精品影院6| 两个人看的免费小视频| 国产欧美日韩精品亚洲av| 欧美一级a爱片免费观看看 | 757午夜福利合集在线观看| 91av网站免费观看| 欧美激情久久久久久爽电影| 国产精品 国内视频| 岛国在线观看网站| 男女视频在线观看网站免费 | 久9热在线精品视频| 色哟哟哟哟哟哟| 真人一进一出gif抽搐免费| 亚洲中文字幕一区二区三区有码在线看 | 亚洲人成电影免费在线| 午夜福利高清视频| 久久久久性生活片| 成人三级黄色视频| 校园春色视频在线观看| 日本 av在线| 法律面前人人平等表现在哪些方面| 五月伊人婷婷丁香| 欧洲精品卡2卡3卡4卡5卡区| 久久精品国产清高在天天线| 淫妇啪啪啪对白视频| 中文字幕人妻丝袜一区二区| 99精品在免费线老司机午夜| av有码第一页| 一级毛片高清免费大全| 亚洲狠狠婷婷综合久久图片| 两性午夜刺激爽爽歪歪视频在线观看 | 亚洲天堂国产精品一区在线| 亚洲国产精品999在线| 一进一出抽搐动态| 国产伦一二天堂av在线观看| 波多野结衣巨乳人妻| 搡老岳熟女国产| 一级作爱视频免费观看| 757午夜福利合集在线观看| 天堂√8在线中文| 欧美日韩福利视频一区二区| 久久久久久久久中文| 香蕉av资源在线| 黄片大片在线免费观看| 丁香六月欧美| 日韩欧美精品v在线| 又大又爽又粗| 日日摸夜夜添夜夜添小说| 精品人妻1区二区| 日日干狠狠操夜夜爽| 桃色一区二区三区在线观看| 午夜两性在线视频| 中文字幕人妻丝袜一区二区| 欧美丝袜亚洲另类 | 国产精品1区2区在线观看.| 免费看十八禁软件| 亚洲成人久久性| 三级毛片av免费| 免费看a级黄色片| 成年女人毛片免费观看观看9| 巨乳人妻的诱惑在线观看| 熟女少妇亚洲综合色aaa.| 亚洲片人在线观看| 国产aⅴ精品一区二区三区波| 丰满人妻一区二区三区视频av | 国产麻豆成人av免费视频| 99国产极品粉嫩在线观看| 性欧美人与动物交配| 日韩 欧美 亚洲 中文字幕| 欧美国产日韩亚洲一区| 免费高清视频大片| 波多野结衣巨乳人妻| 国产精品精品国产色婷婷| 亚洲自偷自拍图片 自拍| 欧美zozozo另类| 小说图片视频综合网站| av福利片在线| 久久国产乱子伦精品免费另类| 婷婷丁香在线五月| 18禁国产床啪视频网站| 国产av麻豆久久久久久久| 最近视频中文字幕2019在线8| 欧美日韩国产亚洲二区| 亚洲一码二码三码区别大吗| 夜夜看夜夜爽夜夜摸| 国产真实乱freesex| 男人舔奶头视频| 久久精品国产99精品国产亚洲性色| 日本五十路高清| 成人三级做爰电影| 欧美午夜高清在线| 国产1区2区3区精品| 久久人妻福利社区极品人妻图片| 欧美日韩精品网址| 后天国语完整版免费观看| 波多野结衣高清无吗| 免费高清视频大片| 亚洲成人久久爱视频| 欧美日本亚洲视频在线播放| 国产成人啪精品午夜网站| 亚洲成人国产一区在线观看| 黄色毛片三级朝国网站| 男女之事视频高清在线观看| 女同久久另类99精品国产91| 亚洲人与动物交配视频| www日本黄色视频网| 级片在线观看| 琪琪午夜伦伦电影理论片6080| 国产成+人综合+亚洲专区| 亚洲国产精品成人综合色| 中文亚洲av片在线观看爽| 久99久视频精品免费| 午夜福利18| 日本在线视频免费播放| 国产黄a三级三级三级人| 欧美乱码精品一区二区三区| 日本 欧美在线| 日本黄大片高清| aaaaa片日本免费| 亚洲成人中文字幕在线播放| 精品第一国产精品| 精品久久久久久久毛片微露脸| 欧美精品啪啪一区二区三区| 18禁裸乳无遮挡免费网站照片| 日韩免费av在线播放| 欧美中文日本在线观看视频| 99久久精品热视频| 黄色毛片三级朝国网站| 成人午夜高清在线视频| 国产一级毛片七仙女欲春2| 免费在线观看成人毛片| 男女做爰动态图高潮gif福利片| 亚洲欧美日韩高清在线视频| 香蕉丝袜av| 小说图片视频综合网站| 国产成人av教育| 中文字幕人成人乱码亚洲影| 50天的宝宝边吃奶边哭怎么回事| 免费看美女性在线毛片视频| 女同久久另类99精品国产91| 久久久精品国产亚洲av高清涩受| 久久精品夜夜夜夜夜久久蜜豆 | 午夜a级毛片| 日韩大尺度精品在线看网址| 国产伦人伦偷精品视频| 亚洲中文字幕日韩| 桃色一区二区三区在线观看| 一进一出好大好爽视频| 欧美高清成人免费视频www| 日日干狠狠操夜夜爽| 国产午夜福利久久久久久| 午夜激情福利司机影院| av在线播放免费不卡| 久久热在线av| 免费看美女性在线毛片视频| 午夜两性在线视频| av片东京热男人的天堂| 国产91精品成人一区二区三区| 男男h啪啪无遮挡| 免费搜索国产男女视频| 日韩免费av在线播放| 亚洲av电影不卡..在线观看| 久久久水蜜桃国产精品网| 亚洲va日本ⅴa欧美va伊人久久| 男插女下体视频免费在线播放| 午夜免费成人在线视频| 正在播放国产对白刺激| 舔av片在线| 精品不卡国产一区二区三区| 无人区码免费观看不卡| 99久久精品热视频| 人人妻人人看人人澡| 一级片免费观看大全| 每晚都被弄得嗷嗷叫到高潮| 老司机福利观看| 长腿黑丝高跟| 又黄又爽又免费观看的视频| 国产真实乱freesex| 国产一区在线观看成人免费| 亚洲 欧美一区二区三区| 最近最新中文字幕大全免费视频| 国产99白浆流出| 久久草成人影院| 亚洲精品美女久久久久99蜜臀| 99国产精品一区二区三区| 午夜精品在线福利| 18禁黄网站禁片免费观看直播| 久久热在线av| 18禁美女被吸乳视频| 色哟哟哟哟哟哟| 91麻豆av在线| 久久香蕉国产精品| av免费在线观看网站| 非洲黑人性xxxx精品又粗又长| 国产蜜桃级精品一区二区三区| 日韩欧美在线乱码| a在线观看视频网站| 婷婷精品国产亚洲av在线| 久久伊人香网站| or卡值多少钱| 久久香蕉精品热| 成人国语在线视频| 免费高清视频大片| 老熟妇乱子伦视频在线观看| 热99re8久久精品国产| 免费av毛片视频| 搡老熟女国产l中国老女人| 在线观看午夜福利视频| 午夜福利18| 亚洲国产精品久久男人天堂| 黄片小视频在线播放| www.熟女人妻精品国产| 一本大道久久a久久精品| 精品免费久久久久久久清纯| 国产精品1区2区在线观看.| 一边摸一边抽搐一进一小说| 国产精品久久久久久久电影 | 久久精品91蜜桃| 两性夫妻黄色片| 少妇粗大呻吟视频| 1024手机看黄色片| 波多野结衣高清无吗| 又爽又黄无遮挡网站| 在线观看一区二区三区| svipshipincom国产片| 他把我摸到了高潮在线观看| 成人18禁高潮啪啪吃奶动态图| 天堂√8在线中文| 看黄色毛片网站| 最近视频中文字幕2019在线8| 欧美性长视频在线观看| 欧美黑人巨大hd| 国产亚洲精品久久久久久毛片| 成人手机av| 身体一侧抽搐| 午夜激情福利司机影院| 五月伊人婷婷丁香| 国产精品 欧美亚洲| 黄片小视频在线播放| 国产激情偷乱视频一区二区| 日韩欧美免费精品| 色老头精品视频在线观看| 99久久无色码亚洲精品果冻| 老司机在亚洲福利影院| 国模一区二区三区四区视频 | 亚洲人成网站在线播放欧美日韩| 日本一二三区视频观看| 91字幕亚洲| 少妇的丰满在线观看| 国产精品久久久久久精品电影| 中文字幕久久专区| 丝袜美腿诱惑在线| 亚洲精品一卡2卡三卡4卡5卡| 亚洲色图 男人天堂 中文字幕| 最好的美女福利视频网| 国产成人啪精品午夜网站| 亚洲精品色激情综合| 9191精品国产免费久久| 欧美成人性av电影在线观看| 亚洲人成网站在线播放欧美日韩| 操出白浆在线播放| 亚洲成人中文字幕在线播放| 最近最新中文字幕大全电影3| 国产精品九九99| 日本熟妇午夜| 午夜免费激情av|