硫杆菌和元素硫在治理重金属污染中的应用研究进展

硫杆菌和元素硫在治理土壤和沉积物中重金属已经有了较多的研究。本文论述了利用硫杆菌和元素硫治理土壤和沉积物中的重金属污染的应用进展。主要包括了利用硫杆菌和元素硫去除重金属的优缺点及主要的影响因素,如温度、pH、细菌种类、元素硫浓度等。     

鸡粪好氧堆肥过程中氨氧化古菌群落结构的动态变化

应用聚合酶链式反应-变性梯度凝胶电泳（PCR DGGE）技术,研究了好氧堆肥过程氨氧化古菌（ammonia oxidizing archaea, AOA）的群落结构和多样性变化。结果表明,不同堆肥时期鸡粪好氧堆肥AOA菌群的群落结构发生了明显的变化。与AOA HH 2(GU2258721)亲缘关系较近的b条带(相似性96%)和未培养泉古菌属［uncultured crenarchaeote NM 152(HQ8752251)］的m条带(相似性99%)是堆肥过程一直存在的AOA菌属。条带c、 b、 f、 i和条带m、 k、 l、 n代表的种属分属两个大类。AOA群落的Shannon Weiner指数（H）、 均匀度指数（EH）均表现为第30 d＞第5 d＞第25 d第45 d＞第3 d第12 d＞第1 d第15 d。对AOA种群数据与不同时期堆体温度、 pH、 全氮、 铵态氮、 亚硝态氮和硝态氮等环境因子的冗余分析表明,堆体温度、 全氮、 亚硝态氮和硝态氮对氨氧化古菌群落演替有着显著的影响（P0.05）,AOA的群落结构在堆肥第1 d、 第5 d、 第25 d、 第30 d、 第45 d变化较大。说明这些参数能有效控制鸡粪好氧堆肥过程中AOA的群落结构。     

茶园一株异养硝化-好氧脱氮菌的鉴定及特性研究

经过菌株富集培养、 分离纯化等一系列步骤,从山东茶园土壤中筛选出一株菌株F1,通过对其进行16S rRNA测序及生理生化指标的测定,结合负染电镜观察结果,参照《伯杰细菌鉴定手册》鉴定该菌株属于黄杆菌属。同时对其硝化和反硝化脱氮能力进行了研究。结果表明,菌株F1能在利用有机物的同时进行硝化和反硝化脱氮作用。经过60 h的培养,在硝化培养基上对氨氮的去除率达到84.54%,对亚硝酸盐的去除率达到91.87%; 在反硝化培养基上对硝酸盐的去除率达到79.17%,对亚硝酸盐的去除率达到84.30%; 在以铵态氮为初始氮源的条件下脱氮能力最强。     

土壤硫的氧化还原及其环境生态效应

硫在土壤中的氧化还原不仅直接影响其在水、大气、生物、岩石圈的交换和循环,而且对生态环境产生影响。综述了土壤中发生的硫的氧化还原过程及硫的氧化还原过程产生的环境生态影响,最后展望了硫的氧化还原及其相关过程的研究前景。     

一株棒状杆菌的氨氧化活性研究

自鸡粪好氧发酵过程中筛选出一株棒状杆菌属(Corynebacterium)的高效异养氨氧化细菌。为明确它的氨氧化活性及其它氮转化特性,研究了初始有机物浓度、氨态氮浓度、pH值、C/N、溶解氧含量、培养温度等环境条件对其氨氧化活性的影响;同时还测定了氨化、硝化和反硝化等氮转化特性。结果显示,该菌株在自养条件下氧化氨态氮仅生成亚硝态氮不生成硝态氮,而在异养条件下氧化氨态氮能同时生成亚硝态氮和硝态氮,且氨态氮利用和亚硝态氮、硝态氮生成总是同步进行的。该菌株氨氧化最适环境条件是有机物浓度680 mg/L、氨态氮浓度424 mg/L、pH值7、C/N 1.77、培养温度35℃和高溶氧量;该菌株除了异养氨氧化特性外,还同时具有氨化、硝化和反硝化等多项氮转化能力。     

元素硫和双氰胺对菜地土壤铵态氮硝化抑制协同效应研究

采用好气培养法,研究了双氰胺（DCD）、元素硫（S0）和元素硫分解中间物（S2O32-）及其组合对蔬菜地土壤氮素硝化抑制作用。结果表明,在培养试验72 d内,DCD+S0、DCD、DCD+ Na2S2O3处理土壤NH4+-N总量分别是N处理的5. 8、5.1、5.9倍;S0、Na2S2O3处理分别是N处理的1.8、1.4倍;而所有硝化抑制剂（DCD、S0、S2O32-）处理土壤NO3--N含量显著低于N处理,表明DCD、S0和S2O32-均能抑制菜地土壤铵态氮硝化。培养试验开始8 d后,Na2S2O3和DCD对铵态氮硝化抑制产生协同效应,16 d后S0和DCD对铵态氮硝化抑制也产生协同效应,这可能是由于S0 氧化中间体S2O32-、S4O62-具有抑制DCD降解作用,延长了DCD硝化抑制作用时间。建议蔬菜生产上推荐使用DCD+S0组合,以提高氮素利用率。     

O_3浓度升高对麦田土壤氨氧化细菌、氨氧化古菌和硝化细菌数量的影响

硝化作用在氮循环过程中至关重要,包括氨氧化作用和亚硝酸盐氧化作用,通过氨氧化反应和亚硝酸盐氧化反应将N素转化为植物可利用的NO;形态。利用开顶式臭氧气室（OTCs,open-topchambers）试验平台,通过大田模拟熏气试验,结合Real-timePCR探讨大气O,浓度升高对麦田土壤氨氧化细菌（AOB）、氨氧化古菌（AOA）及硝化细菌（NOB）数量的影响。结果表明,AOB、AOA和NOB对O,胁迫的反应不一样,AOB基因拷贝数基本上随着O,浓度的升高呈现出降低的趋势,而AOA和NOB基因拷贝数随O_3浓度的升高变化不明显。冬小麦拔节期,当O_3浓度为40、80、120nmol·mol。时,20-40em土层的AOB基因拷贝数分别比对照处理降低39．8％、51．2％和59．4％。AOB和NOB基因拷贝数灌浆期多于收获期,0-10cm土层多于10-20em。AOA基因拷贝数随季节的变化不大。O_3胁迫可通过影响AOB、AOA和NOB的数量和活性来影响土壤的硝化反应,从而影响土壤的氮素循环过程。     

嗜热复合菌对堆肥品质及微生物群落演替的影响

  【目的】  研究嗜热复合菌对畜禽粪污堆肥理化特性和腐熟度的影响,探讨嗜热菌影响堆肥过程的微生物机制。  【方法】  堆料由75%羊粪和25%养鸡发酵床垫料构成,初始原料C/N为28,堆料量1.2 t,高度70～90 cm,开放条垛式堆沤。处理组为堆肥添加0.1%嗜热菌B. fordii FJAT-51578和U. thermosphaericus FJAT-51579等比混合的发酵液,对照组为添加1%市售枯草芽孢杆菌堆肥菌剂(Bacillus subtilis)。堆肥时间为2021年9月18日—10月14日,每两天检测1次温度。堆肥前15天,每两天进行一次翻抛,后期每5天进行一次翻抛,保持堆肥含水量50%～60%,直至高温期结束。在堆肥开始后第1、9和26天取堆肥样品,分析氮磷含量、硝化指数和种子发芽指数。结合扩增子测序,分析堆肥细菌群落结构变化,并揭示其主要环境影响因子。采用PICRUSt分析堆肥有效氮和有效磷代谢的微生物机制。  【结果】  嗜热复合菌添加促进堆肥硝化指数的降低和种子发芽指数的升高,促进堆肥腐熟;堆肥产物碱解氮和有效磷的含量分别比市售菌剂组高11.8%和7.7%。同时,嗜热复合菌的添加改变了细菌群落的分布,降低了堆肥细菌的多样性和丰富度,提高了糖单胞菌、链霉菌和嗜热葡萄孢菌等降解菌的丰度。RDA分析表明,pH和C/N是影响堆肥微生物群落多样性的主要因素,碱解氮与芽孢杆菌和糖单胞菌属丰度正相关,有效磷与嗜热裂孢菌、直丝菌属和马杜拉放线菌属丰度正相关。氮、磷代谢相关京都基因和基因组百科全书同源基因(KO)的PICRUSt分析显示,微生物氮磷循环相关KO的丰度随着堆肥进程均有所增加。添加嗜热菌剂提高了氨化、铵同化、硝酸盐同化、同化/异化硝酸盐还原等氮循环相关KO,及无机磷溶解、酸性磷酸酶和碱性磷酸酶等磷循环相关KO。  【结论】  在畜禽粪污堆肥中添加嗜热复合菌剂加快并延长了高温期,降低了C/N,提高了堆肥中碱解氮和有效磷含量,其中C/N、硝化指数和GI指数等指标在堆肥中期达腐熟程度标准,促进堆肥腐熟。堆肥中添加嗜热复合菌剂增加了细菌氮磷代谢相关KO的表达,提高了腐熟中期堆肥中嗜热菌的丰度和种类,碱解氮与芽孢杆菌和糖单胞菌属丰度呈正相关,有效磷与嗜热裂孢菌、直丝菌属和马杜拉放线菌属丰度正相关。因此,添加嗜热复合菌促进了堆肥有效氮磷的含量。     

围垦对崇明东滩湿地全程氨氧化微生物的影响

全程氨氧化细菌(complete ammonia oxidizers,Comammox)的发现根本改变了学术界对硝化过程的认识,但其地理分异规律及对氮转化过程的贡献仍不清楚。本研究选择长江口崇明东滩不同围垦年限(0、27、51、86 a)稻田表层耕作土壤,采用好氧培养试验测定土壤硝化潜力;通过标靶功能基因amo A实时荧光定量硝化微生物的数量变异特征,包括全程氨氧化细菌(Comammox)、氨氧化细菌(ammonia-oxidizing bacteria,AOB)和氨氧化古菌(ammonia-oxidizing archaea,AOA)。结果表明,与围垦0 a的自然滩涂湿地相比,围垦27、51、86 a的水稻土净硝化速率从N 2.24 mg/(kg·d)分别增加至N 19.3、11.6和11.4 mg/(kg·d),增幅高达5.1倍～8.7倍。AOA的丰度与围垦年限显著正相关。自然滩涂湿地中AOA和AOB的数量分别为0.34×10⁷ copies/g和1.14×10⁷ copies/g,围垦86 a后增幅最高可达27.9倍。自然滩涂湿地中Co...     

Elemental Sulfur Oxidation by Thiobacillus spp. and Aerobic Heterotrophic Sulfur-Oxidizing Bacteria

The prediction of the oxidation rate of elemental sulfur (S 0 ) is a critical step in sulfur (S) fertilizer strategy to supply plant-available sulfur. An incubation experiment was conducted to determine the rate and amount of S 0 oxidation in relation to the contribution of Thiobacillus spp. and aerobic heterotrophic S-oxidizing bacteria. After 84 days, 16.3% and 22.4% of the total S 0 applied to the soil were oxidized at 20 and 30-C, respectively. The oxidation of S 0 proved to be a two-step process with a rapid oxidation during the first 28 days and a slow oxidation from then on. The highest oxidation rate of 12.8 μg S cm-2 d-1 was measured during the first two weeks at 30-C. At 20-C the highest oxidation rate of 10.2 μg S cm-2 d-1 was obtained from two to four weeks after start of the experiment. On an average the soil pH declined by 3.6 and 4.0 units after two weeks of experiment. At the same time the electric conductivity increased nine times. With the oxidation of S 0 the population of Thiobacillus spp. and aerobic heterotrophic S-oxidizing bacteria increased. The corresponding values for Thiobacillus spp. and aerobic heterotrophic S-oxidizing bacteria increased from 2.9 × 10 5 and 1.4 × 10 5 g-1 soil at the start of the experiment to 4 × 10 8 and 5.6 × 10 8 g-1 soil 14 days after S 0 application, respectively. No Thiobacillus spp. was present eight weeks after S 0 application. The results suggested that oxidation of residual S 0 completely relied on aerobic heterotrophic S-oxidizing bacteria.     

Influence of nitrogen and elemental‐sulfur fertilization on sulfur oxidation and mineralization in relation to soil moisture on a calcareous soil of the Inner Mongolia steppe of China

Grassland‐livestock farming is the main agricultural activity in the Inner Mongolia steppe of China. It has been estimated that more than 80% of the grasslands suffer from sulfur (S) deficiency in this region. In an incubation study and a greenhouse experiment with alfalfa, the influence of soil moisture (40% and 70% water‐holding capacity, WHC), nitrogen (0 and 200 mg N (kg soil)^–1 as NH₄NO₃), and elemental sulfur (eS; 0 and 300 mg S (kg soil)^–1) amendments on the apparent eS oxidation, eS‐oxidation rate, net S‐mineralization rate, and S uptake of alfalfa were studied. After 28 d of incubation, the eS‐oxidation rate was four times higher at 70% than at 40% WHC if no N was applied. With N application, soil moisture had only minor effects on eS oxidation during the whole incubation period. In the greenhouse experiment, lower values for eS‐oxidation rate and net S‐mineralization rate were found if no N was applied. Application of N and eS significantly increased alfalfa growth and S uptake. The results of both experiments suggest that combined N and eS applications are the best way to alleviate S deficiency on these calcareous soils.     

使用便携式X射线荧光（PXRF）法原位区分酸性和非酸性冻土带

Frozen soils or those with permafrost cover large areas of the earth’s surface and support unique vegetative ecosystems. Plantsgrowing in such harsh conditions have adapted to small niches, which allow them to survive. In northern Alaska, USA, both moistacidic and non-acidic tundra occur, yet determination of frozen soil pHs currently requires thawing of the soil so that electrometric pHmethods can be utilized. Contrariwise, a portable X-ray fluorescence (PXRF) spectrometer was used in this study to assess elementalabundances and relate those characteristics to soil pH through predictive multiple linear regressions. Two operational modes, SoilMode and Geochem Mode, were utilized to scan frozen soils in-situ and under laboratory conditions, respectively, after soil sampleswere dried and ground. Results showed that lab scanning produced optimal results with adjusted coefficient of determination (R²) of0.88 and 0.33 and root mean squared errors (RMSEs) of 0.87 and 0.34 between elemental data and lab-determined pH for Soil Modeand Geochem Mode, respectively. Even though the presence of ice attenuated fluoresced radiation under field conditions, adjusted R² and RMSEs between the datasets still provided reasonable model generalization (e.g., 0.73 and 0.49 for field Geochem Mode). Principalcomponent analysis qualitatively separated multiple sampling sites based on elemental data provided by PXRF, reflecting differences inthe chemical composition of the soils studied. Summarily, PXRF can be used for in-situ determination of soil pH in arctic environments without the need for sample modification and thawing. Furthermore, use of PXRF for determination of soil pH may provide highersample throughput than traditional eletrometric-based methods, while generating elemental data useful for the prediction of multiplesoil parameters.     

腐植酸和硫酸铁配施改良偏碱烟田土壤的研究

采用田间试验方法,在偏碱烟田中施用腐植酸以及配施腐植酸+硫酸铁,分析其对土壤pH、土壤肥力和烟草农艺性状的影响,研究腐植酸及与硫酸铁配施改良偏碱烟田土的效果.结果表明:(1)腐植酸以及腐植酸+硫酸铁配施均能降低烟田土壤的pH,使pH分别比对照降低0.06,0.21个单位,二者配施的效果比单施腐植酸更好.(2)腐植酸以及腐植酸+硫酸铁配施均对提高土壤肥力有一定作用,腐植酸使土壤碱解氮含量比对照提高9.0％,腐植酸+硫酸铁配施使有机质、碱解氮和有效铁含量分别比对照提高19.6％,10.5％和41.7％,腐植酸+硫酸铁配施使烟叶铁含量比对照提高13.0％.(3)腐植酸以及腐植酸+硫酸铁配施均对烟草的株高、最大叶长、最大叶宽、有效叶片数和干重影响不大,对烟草的生长未产生明显影响.     

不同含水量天然硫酸盐盐渍土中二氧化碳和氧化亚氮的排放

Soil salinization may negatively affect microbial processes related to carbon dioxide(CO_2) and nitrous oxide(N_2O) emissions. A short-term laboratory incubation experiment was conducted to investigate the effects of soil electrical conductivity(EC) and moisture content on CO_2 and N_2O emissions from sulfate-based natural saline soils. Three separate 100-m long transects were established along the salinity gradient on a salt-affected agricultural field at Mooreton, North Dakota, USA. Surface soils were collected from four equally spaced sampling positions within each transect, at the depths of 0–15 and 15–30 cm. In the laboratory, artificial soil cores were formed combining soils from both the depths in each transect, and incubated at 60% and 90% water-filled pore space(WFPS) at 25?C. The measured depth-weighted EC of the saturated paste extract(EC_e) across the sampling positions ranged from 0.43 to 4.65 dS m~(-1). Potential nitrogen(N) mineralization rate and CO_2 emissions decreased with increasing soil EC_e, but the relative decline in soil CO_2 emissions with increasing ECe was smaller at 60% WFPS than at 90% WFPS. At 60% WFPS, soil N_2O emissions decreased from 133 μg N_2O-N kg~(-1) soil at EC_e 0.50 dS m~(-1) to 72 μg N_2O-N kg~(-1) soil at EC_e = 4.65 dS m~(-1). In contrast, at 90% WFPS,soil N_2O emissions increased from 262 μg N_2O-N kg~(-1) soil at EC_e = 0.81 dS m~(-1) to 849 μg N_2O-N kg~(-1) soil at EC_e = 4.65 dS m~(-1), suggesting that N_2O emissions were linked to both soil ECe and moisture content. Therefore, spatial variability in soil EC_e and pattern of rainfall over the season need to be considered when up-scaling N_2O and CO_2 emissions from field to landscape scales.     

苋菜对钾素吸收和利用上的基因型差异

The kinetics of K^ absorption and the utilization of both applied K and K in a calcareous alluvisol with low available K by different genotypes of grain amaranth(Amaranthus spp.),an ancient pseudo-cereal that produces a large biomass and a rich source of health nutrients and mineral elements,were studied by means of ion depletion technique and pot experiments.Grain amaranth had a high affinity for K^ absorption comparison with corn and cotton.However,there were differences among Amaranthus spp.in Kabsorption and utilization.Some cultivars such as R104,CX-4,Du001,Hy015 and Hr029 which had a higher rate of K absorption and stronger affinity for K^ were typical varieties with high K-use efficiency(KUE).The results showed that high KUE cultivars grew quickly,possessed stronger ability to take up soil slowly available K and Mineral K,and did not respond to K fertilization in the soil with low available K.Correspondingly,grain amaranth cultivars CX-77,Cr024,Vd001,Re003 and Sn003 were relatively low in KUE,Compared with high KUE cultivars,they took up more soil available K and both of their dry matter accumulation and K uptake responded to K applied significalty.     

镉胁迫对菊芋生理变化及镉富集的影响

通过用耐镉性较好的徐州菊芋和耐镉性较弱的潍坊菊芋进行对比试验,设置土培CK(Cd~(2+) 0 mmol/L,即对照组)、T1(Cd~(2+) 0.1 mmol/L)、T2(Cd~(2+) 0.5 mmol/L)和T3(Cd~(2+) 1.0 mmol/L)4个处理组,探讨菊芋镉胁迫下各项生理指标和根际土壤指标以及富集能力的影响。结果表明:经过不同镉浓度胁迫21天后,2个菊芋品种的根长、株高、叶长、叶宽均受到相应程度的抑制,抗氧化酶活性随镉浓度的增加而减弱;但丙二醛(MDA)含量变化有所不同,潍坊菊芋的MDA含量伴着镉胁迫加深而增加,徐州菊芋只在T3镉浓度下才出现明显的上升变化,并在21天时达到峰值,为对照组的3.52倍。根际土壤pH大致呈现出持续下降趋势,镉处理20天时潍坊菊芋和徐州菊芋根际土壤pH均在T3处理下达到最低,降幅分别为3.85%和3.41%;有机质含量却表现出不断增加的状态,T3镉处理至第20天时,潍坊菊芋最大增幅为38.60%,徐州菊芋为36.01%;随着施加镉浓度的变大,抑制了菊芋对土壤碱解氮的吸收,总体上潍坊菊芋受抑制程度大于徐州菊芋。2个菊芋品种对镉的富集效果也不同,徐州菊芋各器官镉富集量均高于潍坊菊芋;在转运镉方面,潍坊菊芋根部的镉均等地转移到了叶、茎上,徐州菊芋则将大部分从根部转移到茎上。综上所述,菊芋在镉胁迫下体现出一定的抗性,具有较强镉富集能力,徐州菊芋镉富集能力更为明显。因此,将能源植物菊芋应用于镉污染土壤兼具经济和生态意义。     

接种含ACC脱氨酶的假单胞菌能部分缓解干旱胁迫对豌豆生长、产量及成熟的影响

Two preselected plant growth promoting rhizobacteria （PGPR） containing 1-aminocyclopropane-1-carboxylate （ACC）- deaminase （EC 4.1.99.4） were used to investigate their potential to ameliorate the effects of drought stress on growth, yield, and ripening of pea （Pisum sativum L.）. Inoculated and uninoculated （control） seeds of pea cultivar 2000 were sown in pots （four seeds pot-1） and placed in a wire house. The plants were exposed to drought stress at different stages of growth （vegetative, flowering, and pod formation） by skipping the respective irrigation. Results revealed that inoculation of peas with PGPR containing ACC-deaminase significantly decreased the ＂drought stress imposed effects＂ on the growth and yield of peas. Exposure of plants to drought stress at vegetative growth stage significantly decreased shoot growth by 41% in the case of uninoculated plants, whereas, by only 18% in the case of inoculated plants compared to nonstressed uninoculated control.
Grain yield was decreased when plants were exposed to drought stress at the flowering and pod formation stage, but inoculation resulted in better grain yield （up to 62% and 40% higher, respectively） than the respective uninoculated nonstressed control. Ripening of pods was also delayed in plants inoculated with PGPR, which may imply decreased endogenous ethylene production in inoculated plants. This premise is further supported by the observation that inoculation with PGPR reduced the intensity of classical ＂triple＂ response in etiolated pea seedlings, caused by externally applied ACC. It is very probable that the drought stress induced inhibitory effects of ethylene could be partially or completely eliminated by inoculation with PGPR containing ACC-deaminase.     

根结线虫接种对黄瓜植株根际土壤pH和微生物的影响

通过人工接种不同数量的根结线虫卵,测定了黄瓜植株根际土壤的pH和微生物数量。结果表明,随接种数量的增加,根际土壤好气性细菌数量、厌气性细菌数量、细菌总数及细菌/真菌(B/F)逐渐降低;真菌数量却逐渐升高;放线菌数量在接种量为2000个·株-1时显著升高,之后随着接种量的增加逐渐降低;放线菌/真菌(A/F)在接种量为2000个·株-1时略有升高,之后随着接种量的增加逐渐降低。接种根结线虫后,黄瓜植株根际土壤中好气性细菌数量和B/F值与pH呈显著正相关,厌气性细菌数量、细菌总数、放线菌数量以及A/F值与pH呈极显著正相关,真菌数量与pH呈显著负相关。根结线虫侵染黄瓜植株导致根际土壤发生"真菌化",显示土壤质量下降。     

Quantitative real-time PCR effectively detects and quantifies colonization of sclerotia of Sclerotinia sclerotiorum by Trichoderma spp.

Some members of the fungal genus Trichoderma are able to colonize and destroy sclerotia, the thick-walled resting structures of the soilborne plant pathogenic fungus Sclerotinia sclerotiorum, thus providing a potential means of biological disease control. However, current methods to detect and quantify colonization of sclerotia are labor-intensive, and generally qualitative rather than quantitative in nature. Our objective was to develop quantitative real-time PCR (polymerase chain reaction) methods to detect and measure colonization of sclerotia by Trichoderma spp. Specific PCR primer/probe sets were developed for Trichoderma spp. and for S. sclerotiorum. A total of 180 ITS1 (internal transcribed spacer) and ITS2 sequences from different species in the genus Trichoderma were aligned, and consensus sequences were determined. Six candidate primer sets were based on conserved regions of the consensus sequence, and the specificity of each nucleotide sequence was examined using BLAST (Basic Local Alignment Search Tool; NCBI) software. Each candidate primer set was tested on genomic DNA of T. harzianum strain ThzID1-M3, as well as six different Trichoderma isolates from soil, and on genomic DNA of S. sclerotiorum. The optimum primer/probe set selected, TGP4, successfully amplified genomic DNA of all Trichoderma isolates tested, and showed high precision and reproducibility over a linear range of eight orders of magnitude, from 85 ng to 8.5 fg of T. harzianum genomic DNA. TGP4 did not amplify S. sclerotiorum DNA. A specific PCR primer/probe set (TMSCL2) was developed for S. sclerotiorum, based on the calmodulin gene sequence. TMSCL2 did not amplify Trichoderma DNA. Quantitative real-time PCR with the primers then was evaluated in experiments to test differential effects of two soil moisture levels (−50 kPa, −1000 kPa matric potential) on biocontrol of S. sclerotiorum by indigenous Trichoderma spp. Periodically over 40 days, Trichoderma and S. sclerotiorum DNA in sclerotia were quantified by PCR with appropriate primers. Over 90% of the sclerotia were colonized by indigenous Trichoderma spp. at −1000 kPa, over the 40-day period, compared to only 60% at −50 kPa. In addition to determining incidence of colonization, the PCR method allowed measurement of the extent of sclerotial colonization, which also was significantly greater in the drier soil. Quantitative real-time PCR with the TGP4 primer/probe set provides a sensitive detection and measurement tool to evaluate colonization of sclerotia by Trichoderma spp.