硫红霉素菌渣有机肥对大豆土壤中耐药菌及相关抗性基因的影响

【目的】研究硫红霉素菌渣有机肥施用后对大豆农田土壤中耐药菌及相关抗性基因丰度的影响,为硫红霉素菌渣有机肥施用生物安全性评价提供科学依据。【方法】检测不同菌渣有机肥施用量下,大豆不同生长时期土壤中耐药细菌的数量和种类,并采用荧光定量PCR方法检测相关抗性基因(ARGs)污染水平,分析菌渣有机肥施用对耐药菌及相关抗性基因的影响。【结果】大豆苗期土壤中各类细菌数量大于结果期,苗期样品中土壤细菌总数、硫红霉素抗性菌数显著高于对照组,而青霉素、头孢拉定抗性菌株数与对照组差异不显著。在结果期样品中,施加了硫红霉素药渣有机肥的土壤中各类菌落总数与对照组差异不显著。所获得14株硫红霉素耐药菌菌株分布于11个菌属,其中假节杆菌属、芽胞杆菌属、类谷氨酸杆菌属菌株数占总菌株数的比例最高,25株青霉素耐药菌菌株分布于7个菌属,其中链霉菌属菌株数最高。12株头孢拉定耐药菌分布于5个菌属,其中假单胞菌属菌株数占比例达到50.00%。施用硫红霉素菌渣有机肥对土壤中常见ARGs的绝对丰度和相对丰度有一定的影响,但影响不显著。【结论】硫红霉素菌渣有机肥施用后对作物土壤中ARGs水平没有显著影响。

生物炭与炭基肥对大豆根际土壤细菌和真菌群落的影响

土壤微生物在农田土壤生态系统中发挥重要作用,然而秸秆生物炭与炭基肥处理对微生物群落的影响以及对农田生态环境的意义尚不清楚。以黄淮海平原豆-麦轮作为研究对象,采用荧光定量PCR和Illumina高通量测序技术比较不同施肥方式对土壤细菌和真菌群落的丰度、组成和多样性差异,探究秸秆还田、生物炭以及炭基肥添加对根际土壤微生物群落结构的影响。试验处理包括单施化肥对照(CK)、秸秆全量还田配施化肥处理(CS)、炭基肥处理(BCF)、低量生物炭配施化肥处理(LB)以及高量生物炭配施化肥处理(HB)。结果表明:与对照CK相比,LB、BCF和HB处理显著提高了根际土壤有机碳和速效磷含量;LB、BCF和HB处理对土壤细菌丰度没有显著影响,HB处理显著降低了变形菌门的相对丰度。HB处理的真菌丰度显著高于CK,增加了86.3%,真菌群落Chao1和ACE指数较CK分别显著增加了5.9%和5.8%;与CK和CS相比,LB、HB和BCF处理明显改变了真菌群落结构。冗余分析(RDA)表明,土壤有机碳、总氮和速效钾是改变真菌群落结构的主要驱动因子。综上,研究表明大豆根际真菌群落对生物炭施加的敏感程度高于细菌,且土壤碳和氮含量是影响真菌和细菌群落结构的关键因子。该研究结果可为秸秆资源的合理利用提供理论参考。

单甲脒农药对大豆-土壤系统的生态影响

研究了单甲脒农药（DMAH）对大豆－土壤系统的生态影响．结果表明：除CK外，5个设置浓度［（DMAH）／mL·L－1］0．25、0．5、2．5、5.0、25均对大豆虫害有不同的防治效果．其中＞2．5时可防治大豆多种虫害；大豆对DMAH稀释液的pH值反应敏感，高浓度处理的叶片因酸度大而出现药害；推荐浓度［（DMAH）＝0．25mL·L－1］，对植株的生长和生理功能有促进作用．可使总生物量及产量分别增加20％左右，且籽粒中农药无残留，土壤农药残留量也小；当处理pH值＜5时会对叶片有伤害．生理功能被抑制，总生物量及产量都不如CK；DMAH在植物器官及土壤都有不同程度的残留量．其中叶残留量大于籽粒及土壤．可以认为，低浓度DMAH处理不会对植物－土壤系统造成污染．

UV-B增强对土壤-大豆系统CO2排放的影响

采用静态箱-气相色谱法测定CO2排放通量,研究了人工模拟UV-B辐射增强对土壤-大豆系统CO2排放的影响.结果表明,在相同的气象条件和田间管理下,UV-B辐射增强对CO2排放的季节变化模式无明显影响.在植株结荚前,UV-B增强对CO2排放通量没有显著影响,但从植株结荚到成熟,CO2排放通量显著降低,2004和2006年分别降低了49.03%和48.13%;并显著降低了整个生育期的CO2累积排放量,2004和2006年分别降低了43.26%和44.79%.收割实验表明,在分枝开花期,UV-B增强降低了土壤CO2排放,但对植株地上部分的CO2排放没有明显影响;从结荚到成熟,UV-B增强主要通过降低植株地上部分CO2排放通量来降低土壤-大豆系统的CO2排放.UV-B辐射增强降低了大豆生态系统碳固定量,在分枝开花-结荚阶段的固定碳量减少了352.73g/m2CO2,在结荚-鼓粒阶段固定碳量减少了1456.25g/m2CO2.

UV-B增强与秸秆施用对土壤-大豆系统呼吸速率和N_2O排放的影响

采用人工模拟UV-B增强方式,通过大田试验,研究了UV-B增强与秸秆施用对土壤-大豆系统呼吸速率和N2O排放通量的影响.结果表明:在三叶-分枝期、开花-结荚期、鼓粒成熟期和全生长期,UV-B增强,系统平均呼吸速率分别降低了59.88%,65.47%,67.35%和64.44%,N2O平均排放通量分别降低了37.94%,24.61%,48.42%和34.16%;秸秆施用促进了系统呼吸速率,4个时期平均呼吸速率分别增加了59.88%,61.50%,99.16%和64.44%;降低了全生长期的N2O平均排放通量,但没有达到显著差异水平(P=0.236).UV-B增强和秸秆施用复合处理显著增大土壤-大豆系统的呼吸速率,降低全生长期的N2O平均排放通量,但没有达到显著差异水平(P=0.229).

地表UV-B辐射增强对土壤-大豆系统N_2O排放的影响

通过大田试验和室外盆栽试验,采用人工增加紫外辐射的方法模拟UV-B辐射增强,用静态箱-气相色谱法测定N2O排放通量,研究地表UV-B辐射增强对土壤-大豆系统N2O排放的影响。结果表明:在相同的气象条件和田间管理措施下,UV-B辐射增强没有改变土壤-大豆系统N2O排放通量的季节性变化规律。但从植株结荚到成熟,UV-B辐射增强降低了土壤-大豆系统N2O排放通量,进而降低了N2O的累积排放量。收割实验发现,在分枝开花期,UV-B辐射增强对土壤N2O排放影响明显,降低了土壤N2O排放通量;从结荚至鼓粒期,UV-B辐射增强主要通过降低植株地上部分N2O排放通量来降低土壤-大豆系统的N2O排放。UV-B辐射增强显著降低了植株的生物量,并影响到植株的氮代谢和土壤NH4+-N与微生物氮。UV-B辐射增强可能会导致农田生态系统NO排放量降低。

四川地区结瘤大豆根际土壤中紫云英、苜蓿和三叶草根瘤菌的多样性分析

为了解四川地区结瘤大豆根际土壤是否存在与紫云英、苜蓿和三叶草共生结瘤的根瘤菌及其多样性,从四川盆地采集不同种植模式下结瘤大豆根际土壤样品31份,利用紫云英、苜蓿和三叶草进行盆栽捕获试验以获得共生根瘤,从根瘤中分离纯化出根瘤菌,对其进行表型和分子鉴定;并将根瘤菌回接到紫云英、苜蓿、三叶草和大豆根际以检测根瘤菌的寄主范围.在捕获实验中,苜蓿和三叶草分别在6个土壤样品中共生结瘤,而紫云英只在2个土样中共生结瘤,并从共生根瘤中分离出14株根瘤菌;16S r DNA基因序列的相似性分析表明这14株根瘤菌均属于根瘤菌属(Rhizobium),且16S r DNA基因的系统发育树揭示它们分属于根瘤菌属的不同种;这14株根瘤菌在回接实验中都只能让捕获豆科植物结瘤,而不能让其他3种豆科植物结瘤.本研究结果表明,四川地区结瘤大豆根际土壤中的紫云英根瘤菌(Rhizobium astragali)、苜蓿根瘤菌(R.meliloti)和三叶草根瘤菌(R.leguminosarum var.trifolii)资源较少,其与大豆根瘤菌(Bradyrhizobium japonicum)分属于不同的属,且根瘤菌的回接实验进一步证明了根瘤菌的寄主专一性.

Soil Respiration During a Soy bean-Growing Season

Soil respiration induced by soybean cultivation over its entire growing season and the factors influencing soil respiration were investigated to examine the seasonal pattern of soil respiration induced by soybean cultivation, explore soybean growth and photosynthesis on soil respiration, and determine the temperature dependence on soil respiration. Soil respiration in a pot experiment with and without soybean plants was sampled using the static chamber method and measured using gas chromatograph. Air temperature was a dominant factor controlling soil respiration rate in unplanted soil. Additionally, rhizosphere respiration comprised 62% to 98% of the soil respiration rate in the soybean-planted soil varying with the soybean growth stages. Harvesting aerial parts of soybean plant caused an immediate drop in the soil respiration rate at that stage. After harvesting the aerial parts of the soybean plant, a highly significant correlation between soil respiration rate and air temperature was found at the flowering stage （P 〈 0.01）, the pod stage （P 〈 0.01）, and the seed-filling stage （P 〈 0.05）. Thus, rhizosphere respiration during the soybean-growing period not only made a great contribution to soil respiration, but also determined the seasonal variation pattern of the soll respiration rate.

Surfactant Linear Alkylbenzene Sulfonate Effect on Soil Cd Fractions and Cd Distribution in Soybean Plants in a Pot Experiment

A pot experiment was conducted to study the effect of an anionic surfactant linear alkylbenzene sulfonate(LAS) application on cadmium(Cd)fractions in soils and Cd distribution in different tissues of soybean(Glycine max)plants as well as soil pH.Soil samples were treated with three levels of Cd(0,5,and 10 mg kg^(-1))and five levels of LAS(0,5, 15,50,and 100 mg L^(-1)).Results indicated that compared to the control soils(no Cd and no LAS treatment),soil pH increased and available Cd decreased in the soil treated with external Cd and watered with LAS solutions.Meanwhile, soil exchangeable Cd and Cd bound to carbonates decreased;Cd bound to amorphous iron and manganese oxides and Cd bound to organic matter increased.In addition,LAS application could reduce enrichment of Cd in soybean plants, resulting in decreased Cd in the soybean plants.Thus,suitable LAS application could decrease bioavailability and mobility of soil Cd.

Application of Soil Amendments Reduced Cadmium Bioavailability but Restrained Soybean Growth

The soybean is a crop which easily accumulates cadmium(Cd),which threatens human health.To assess the impact of the application of classic soil amendments on the Cd concentration in the soybean and the Cd bioavailability in the soil,a field study was conducted in Xiangtan Country(XT)and Liling City(LL),with inorganic-organic-microbial matter(T1)and silicon-calcium-magnesium oxide from natural minerals(T2)as two soil amendments in this study.The results indicated that the soil pH in the two sites increased significantly,up to 0.7~1.1 units and the Cd concentration in the stem,leaf,husk and seed in the two sites decreased differently.Of which,the soil available Cd in Xiangtan County(XT)decreased by 11.9%~16.0%,the enrichment factor(EF)and translocation factor(TF)reduced by 37.9%and 23.5%,respectively.Both soil amendments were effective in increasing the soil pH,reducing the seed and soil available Cd,but the soil organic matter,total N,stem length and grain yields decreased slightly.In conclusion,the Cd bioavailability was reduced but the soybean growth was restrained with the application of the soil amendments.

Mutant acetolactate synthase （ALS） gene as a selectable marker for Agrobacterium-mediated transformation of soybean

Soybean is one of the crops most difficult to be manipulated in vitro. Although several soybean marker genes, all the selectable markers used were from bacteria origin. To find suitable selectable marker gene from plant origin for soybean transformation, a mutant acetolactate synthase （ALS） gene from Arabidopsis thaliana was tested for Agrobacterium-mediated soybean embryo axis transformation with the herbicide Arsenal as the selective agent. Transgenic soybean plants were obtained after the herbicide se- lection and the To transgenic lines showed resistance to the herbicide at a concentration of 100 g/ha. ALS enzyme assay of To transgenic line also showed higher activity compared to the wild type control plant. PCR analysis of the T1 transgenic lines confirmed the integration and segregation of the transgene. Taken together, our results showed that the mutant ALS gene is a suitable selectable marker for soybean transformation.

Feasibility of Tea Saponin-Enhanced Soil Washing in a Soybean Oil-Water Solvent System to Extract PAHs/Cd/Ni Efficiently from a Coking Plant Site

Mixed contaminated brownfield sites have brought serious risks to human health and environmental safety. With the purpose of removing polycyclic aromatic hydrocarbons （PAHs） and heavy metals from a coking plant site, an innovative technology for ex-situ washing was developed in the present work. The combination of 15.0 mLL-1 soybean oil and 7.5 g L-1 tea saponin proved an effective method to extract co-pollutants from soil. After two consecutive washing cycles, the efficiency rates of removal for 3-, 4-, 5（＋6）-ring, and total PAHs, Cd, and Ni were approximately 98.2%, 96.4%, 92.3%, 96.3%, 94.1%, and 89.4%, respectively. Meanwhile, as evaluated by Tenax extraction method and metal stability indices, the residual PAHs and heavy metals after consecutive washing mainly existed in the form with extremely low bioaccessibility in the soil. Thus, in the soil after two washing cycles, there appeared limited environmental transfer risk of co-pollutants. Moreover, a subsequent precipitation method with alkaline solution and PAH- degrading strain Sphingobium sp. PHE9 inoculation effectively removed 84.6%-100% of Cd, 82.5%-91.7% of Ni, and 92.6%-98.4% of PAHs from the first and second washing solvents. The recovered solvents also exhibited a high recycling effectiveness. Therefore, the combined cleanup strategy proposed in this study proved environmentally friendly, which also played a major role in risk assessment and marlagement in mixed polluted sites.

Heavy Metal Contamination in Soil and Soybean near the Dabaoshan Mine, South China

Concentrations of Pb, Cd, Cu, Zn, Cr and Ni in soybean (Glycine max L.) grown near the Dabaoshan Mine were investigated, and their potential risk to the health of inhabitants was estimated. In the Fandong (FD) and Zhongxin (ZX) villages, which are near the Dabaoshan mineral deposit, concentrations of Pb (0.34 mg kg^(-1) for FD), Cd (0.23 mg kg^(-1) for ZX) and Cr (1.14 and 1.75 mg kg^(-1) for FD and ZX, respectively) in the seeds of soybean exceeded the tolerance limit set by Chinese standards. The estimated daily intakes (EDIs) from consumption of soybean seeds for FD inhabitants were 0.570, 0.170, 38.550, 142.400, 1.910 and 14.530 μg d^(-1) kg^(-1) boby weight for Pb, Cd, Cu, Zn, Cr and Ni, respectively. Our results indicate that soybeans grown in the vicinity of the Dabaoshan Mine accumulate some metals, and the seeds pose a potential health risk to the local inhabitants.

Synergistic Effects of Biochar and NPK Fertilizer on Soybean Yield in an Alkaline Soil

Biochar effects on legume growth and biological nitrogen fixation have been studied extensively, mostly in acidic soils with laboratory produced biochar. In the present study, a pot experiment in a full factorial experimental design was performed to examine soybean yield and nodulation of three genotypes grown with or without biochar and NPK fertilizers in an alkaline soil. We observed synergistic effects of biochar and NPK fertilizer applications on biomass and seed yields for all three soybean genotypes. Total biomass production and seed yield increased on average by 67% and 54%, respectively, with biochar and by 201% and 182% with NPK fertilizer application compared to the control. When applications of biochar and NPK fertilizer were combined, the increases were 391% and367%, respectively. However, the biomass production in the control was very low(692 kg ha-1) due to a high soil p H(8.80). The nodulation increased with biochar and NPK fertilizer applications, and was largest with the combined application. A correlation was found between leaf chlorophyll content(single photon avalanche diode value) and nodule number. We suggested that the synergistic increase in yield was due to a decrease in soil p H caused by biochar and NPK fertilizer applications thereby increasing P availability in this alkaline soil.

Soil Phosphorus Fractions Change in Winter in a Corn-Soybean Rotation with Tillage and Phosphorus Fertilization

Determining how agricultural management practices affect soil phosphorus(P) over the winter may further our understanding of the soil P cycle under specific environmental conditions in eastern Canada. This study assessed changes over winter for soil P fractions and other selected chemical properties as affected by tillage and P fertilization. In 1992, a long-term corn(Zea mays L.) and soybean(Glycine max L.) rotational experiment was established in the province of Quebec, Canada. Soil samples(0–15 cm) were collected in fall 2001 and 2007 after a soybean harvest, and in the following spring 2002 and 2008 before corn seeding, in main plots under moldboard plow and no-till managements and selected subplots fertilized with 0, 17.5, or 35 kg P ha-1and 160 kg N ha-1. Soil samples were analyzed for P fractions and other chemical properties to assess changes over winter for 2001–2002 and 2007–2008. Changes over winter of all soil P fractions were significant for the two periods, indicating the occurrence of soil P transformation and movement over winter. The Mehlich-3-extractable Fe, Al, Ca, and Mg decreased during the two studied periods. Tillage had no significant effect on all soil P fractions. The resin-extractable P in 2001–2002 and Na HCO3-extractable inorganic P and Na OH-extractable organic P during the two winters were significantly increased under P fertilization. This study demonstrated that P in cultivated soils changed during winter as a result of changes in labile P fractions possibly due to the solubilization of residual fertilizer P combined with environmental factors.

Minimum Data Set for Assessing Soil Quality in Farmland of Northeast China

Soil quality assessment provides a tool for agriculture managers and policy makers to gain a better understanding of how various agricultural systems affect soil resources. Soil quality of Hailun County, a typical soybean （Glycine max L. Merill） growing area located in Northeast China, was evaluated using soil quality index （SQI） methods. Each SQI was computed using a minimum data set （MDS） selected using principal components analysis （PCA） as a data reduction technique. Eight MDS indicators were selected from 20 physical and chemical soil measurements. The MDS accounted for 74.9% of the total variance in the total data set （TDS）. The SQI values for 88 soil samples were evaluated with linear scoring techniques and various weight methods. The results showed that SQI values correlated well with soybean yield （r = 0.658＊＊） when indicators in MDS were weighted by the regression coefficient computed for each yield and index. Stepwise regression between yield and principal components （PCs） indicated that available boron （AvB）, available phosphorus （AvP）, available potassium （AvK）, available iron （AvFe） and texture were the main factors limiting soybean yield. The method used to select an MDS could not only appropriately assess soil quality but also be used as a powerful tool for soil nutrient diagnosis at the regional level.