CaCl_2对干旱地区钙质土壤放线菌分离效果的影响

采用稀释平板法研究了高氏1号培养基中加入CaCl2对钙质土壤放线菌分离效果的影响。结果表明:(1)加入CaCl2对干旱区钙质土壤放线菌的出菌量有一定影响,大部分土壤的放线菌总数较对照增长23.9%~47.0%,加钙处理与对照的差异达到显著水平(P0.05);加钙后链霉菌数量有增有减,增、减幅度分别为17.5%~123.3%、18.0%~42.5%,减少幅度未达到显著水平(P0.05)。(2)向高氏1号培养基中加钙有利于广谱高效拮抗性放线菌的分离。     

土壤放线菌的选择性分离及其代谢产物抗菌活性评价*

从土壤中选择性分离放线菌,并进行抗菌活性评价,以期筛选到具有潜在应用开发价值的菌株。以改良的HV培养基为分离培养基,添加萘啶酮酸、制霉菌素作为抑制剂,对从安徽、重庆、河北、湖北、江苏、吉林、山东、四川、天津、云南及德国莱比锡等地采集的45份土壤样品中的放线菌进行了选择性分离。以植物病原菌黑曲霉、棉花枯萎病菌、稻瘟病菌、辣椒疫霉、茄青枯病菌为指示菌,通过平板琼脂块初筛,并结合摇瓶复筛和盆栽防效进行评价。共选择性分离到570株放线菌;初筛显示,212个菌株代谢产物显示了抗真菌或抗细菌活性,占总菌株数量的37.2%,33个菌株代谢产物兼具抗真菌活性和抗细菌活性,占菌株数量的5.8%;摇瓶复筛显示,其中2株放线菌TJ231、TJ430发酵液对黑曲霉、棉花枯萎病菌、稻瘟病菌、辣椒疫霉、茄青枯病菌显示了强烈的抑制效果;1株放线菌TJ561发酵液对黑曲霉、棉花枯萎菌、稻瘟菌、辣椒疫霉显示了强烈的抑制效果;盆栽试验表明,放线菌TJ231、TJ430、TJ561发酵液对辣椒疫霉病的保护效果分别为56.2 %、87.2%、61.4%。     

太白山土壤放线菌分离方法及生态分布规律

研究了改良高氏培养基、甘油-天门冬酰胺培养基、R2A培养基、改良高氏二号培养基、海藻糖-脯氨酸培养基对太白山不同植被土壤放线菌的分离效果,并分析放线菌的生态分布规律。结果表明:太白山南北坡土壤养分大体呈现为中海拔林地含量相对较高,有机质和全氮含量最高分别为77.81 g·kg~(-1)和5.35 g·kg~(-1),低海拔林地与高海拔林地含量相对较低,有机质和全氮含量最低分别为11.77 g·kg~(-1)和0.85 g·kg~(-1),即自低海拔至高海拔不同林地类型土壤养分整体呈现先升高后降低的趋势;甘油-天门冬酰胺培养基分离的放线菌数量与种类均高于其它培养基,其数量最高可达258×10~4cfu·g~(-1)土,种类最多为35种;改良高氏培养基分离南坡的效果优于北坡;R2A培养基分离中高海拔林地土壤优势更明显。中海拔林地类型,如北坡的红皮桦、牛皮桦,南坡的牛皮桦,分离的放线菌数量均显著高于其它林地类型;土壤速效磷和全氮含量对放线菌的分离效果影响较大。     

拮抗农业致病菌放线菌的分离、筛选与鉴定

采用梯度稀释法从土壤中分离出放线菌G5。该菌株对苹果轮纹病菌、油菜菌核病菌等多种农业致病真菌有很强的抑制作用。用16SrDNA序列分析法对G5进行序列分析,该菌株16SrDNA序列与链霉菌属的13个放线菌的同源性为99％。用传统细菌学的方法对G5菌株进行鉴定,其形态特征和生理生化特性与链霉菌的特征和特性相一致,表明该菌株属于链霉菌。     

高山草甸土壤放线菌分离方法研究

采用平板涂抹分离法研究了不同分离培养基及样品预处理对高山草甸土放线菌分离的效果。结果表明：土样在120℃干热处理1 h后其悬浮液加0.05%SDS（十二烷基磺酸钠）和6%酵母膏,40℃振荡30 min,能促进放线菌孢子萌发,增加放线菌的分离数量,并抑制杂菌生长,是高山草甸土放线菌分离时理想的预处理方法。改良HV琼脂和高氏1号加重铬酸钾（50 mg/L）培养基是较好的分离培养基。     

土壤拮抗放线菌的分离及其抗菌活性筛选

以常见植物病原真菌和细菌为指示菌,从分离的422株放线菌中筛选出28株可代谢抗细菌活性物质的菌株,其中N331、N393菌株发酵液对供试12种真菌有显著的抑制作用。离体条件下,N331、N393菌株发酵液对供试真菌菌丝生长的抑制中浓度(EC50)均小于15 mL/L;对玉米弯孢病菌、棉花枯萎病菌、小麦根腐病菌、黄瓜霜霉病菌孢子萌发的EC₅₀亦小于15 mL/L。盆栽试验结果表明,N331、N393菌株发酵原液对黄瓜霜霉病的保护效果分别为97.4%和85.6%,治疗效果分别为83.3%和59.3%。     

土壤稀有放线菌的选择性分离及其抗菌活性研究

在优化土壤稀有放线菌选择性分离方法的基础上,对从安徽、浙江、山东、陕西等地采集的31份土样中的稀有放线菌进行了选择性分离。结果表明:以改进的HV培养基为分离培养基,将土壤样品稀释100倍,添加2×10-5 g/mL重铬酸钾＋2×10-5 g/mL萘啶酮酸＋1×10-5 g/mL卡那霉素作为抑制剂分离效果最好。以常见植物病原真菌和细菌为指示菌,从分离的417株放线菌中筛选出H32、H75、H223、H227、H238等5株可代谢抗菌活性物质的菌株。离体条件下,5株放线菌菌株发酵液对小麦根腐病菌Bipolaris sorokiniana、玉米大斑病菌Exserohilum turcicum、烟草赤星病菌Alternaria alternate的菌丝生长和孢子萌发均有强烈的抑制作用,其中H223和H238菌株发酵液对病原真菌菌丝生长的抑制中浓度(EC50)值均小于10 mL/L;H32菌株发酵液对小麦根腐病菌、玉米大斑病菌、烟草赤星病菌孢子萌发的EC50值分别为25.5、28.9和29.9 mL/L。5株放线菌发酵液对枯草芽孢杆菌、大肠杆菌、金黄色葡萄球菌抑制作用显著,其中H223菌株发酵液抑制作用最强,抑菌圈达30 mm以上。盆栽实验结果表明,5株放线菌发酵液对小麦白粉病的保护效果为73.45％~82.35％,治疗效果为67.74％~70.80％。     

稻草畏土壤残留的生物测定初报

移栽稻田一次性除草剂稻草畏系由稻无草、乙草胺及一些添加剂混配而成,具有高效、广谱、低毒、经济、对水稻安全、适用地域广等特点,可有效地防除移栽稻田一年生和多年生单、双子叶杂草,一次用药即可控制移栽稻田全季草害。稻草畏推出后,深受广大植保、农资部门和农民的欢迎,1992～1994年     

拮抗茄子黄萎病菌土壤放线菌的分离筛选和鉴定

为了研究土壤拮抗性放线菌对茄子黄萎病的生防价值,从陕西省延安市宝塔区与汉中地区土壤中分离、纯化获得154株具有纤维素降解活性的放线菌。采用平板对峙培养法进行初筛,用生长速率法对抑菌效果较好的菌株ZX-10-4进行抑菌谱的测定,并进行盆栽试验验证其对茄子黄萎病的防治效果,通过形态、培养特征及16S rDNA序列分析研究其分类地位。结果显示,有17株放线菌对茄子黄萎病菌抑菌效果较好且抑菌活性稳定,其中菌株ZX-10-4的抑菌活性最高,抑菌率达90.15%;活性菌株ZX-10-4发酵原液对茄子黄萎病的保护和治疗作用分别为70.00%和60.33%,稀释5倍液的防效也在50%以上。根据形态、培养特征、生理生化指标和16S rDNA序列分析,初步确定菌株ZX-10-4为白网链霉菌Streptomyces albireticuli的近似种。     

香蕉枯萎病颉颃放线菌的分离和筛选

从采自内蒙古、海南的土样中经分离、纯化得到64株优势放线菌,采用平板对峙培养法对香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)进行颉颃放线菌的筛选.通过皿内颉颃试验,获得4株抑菌效果较好的颉颃放线菌,以HN3菌株的抑菌效果最为明显,发酵液抑茵试验表明,其抑菌物质存在于菌丝内.     

Evaluation of the bioavailability of the herbicide prosulfocarb through adsorption on soils and model soil colloids, and through a simple bioassay

Adsorption isotherms of prosulfocarb were determined on soils with different physicochemical properties. The extent of adsorption increased with the organic carbon content, but the results also suggested the involvement of some inorganic soil surfaces. In order better to understand the role of each soil surface, adsorption-desorption isotherms were determined on model soil colloids consisting of a soil humic acid, a montmorillonite, a synthetic aluminium hydroxide and their associations. The highest adsorption was observed on the humic acid, but it was also confirmed that the montmorillonite interacted with prosulfocarb. In contrast, the aluminium hydroxide was not active and its association with montmorillonite provoked a decrease in adsorption compared with montmorillonite alone. Except for humic acid, the highest adsorption took place on the ternary association montmorillonite-aluminium hydroxide-humic acid. On each surface tested, the adsorption was largely reversible and decreased at increasing pH. The herbicidal activity of prosulfocarb was tested by the determination of the germination and growth of a typical weed, Lolium multiflorum Lam, in contact with prosulfocarb solutions at different concentrations. The herbicide did not prevent the germination of the seeds but inhibited the growth of the roots and leaves. The same test was performed in the presence of the ternary system to evaluate the influence of adsorption on the plant bioavailability. It was concluded that the presence of an adsorptive surface reduced the herbicidal activity and that the simple bioassay proposed could be useful in predicting the extent of adsorption in a given soil.     

海洋放线菌A3202的分离鉴定及其对柑橘采后病害的防效

为获得用于防治柑橘采后病害的海洋放线菌,以柑橘青霉病菌Penicillium italicum、柑橘绿霉病菌P.digitatum和柑橘炭疽病菌Colletotrichum gloeosporioides为指示菌,对分离到的25株海洋放线菌进行筛选。通过平板对峙法测定拮抗菌株的抑菌谱,并测定其发酵产物粗提物对柑橘果实采后病害的防效,根据形态特征、培养特征、生理生化特征及16S rDNA序列对其进行鉴定。结果表明,分离自鳞笠藤壶Tetraclita squamosa的菌株A3202对3种指示菌均具有强抑菌活性,对供试13种植物病原菌的菌丝生长具有不同程度的抑制作用,尤其对柑橘青霉病菌和柑橘绿霉病菌的抑制作用最强,抑菌带宽度分别可达2.33±0.05 cm和2.32±0.10 cm。该菌株800 mg/L粗提物溶液对接种后7 d的柑橘青霉病、柑橘绿霉病和柑橘炭疽病的相对防效均可达100%。初步鉴定该菌株为放线菌中的米修链霉菌Streptomyces misionensis。     

Lessons from 10 years of stakeholder adoption of a soil bioassay for assessing the risk of spinach Fusarium wilt

Although the maritime Pacific Northwest (PNW) is the only region of the United States suitable climatically for spinach seed production, the acidic soils are highly conducive to spinach Fusarium wilt caused by Fusarium oxysporum f. sp. spinaciae. A soil bioassay developed to quantify the risk of spinach Fusarium wilt in fields has been offered to seed growers annually since 2010. Soil sampled from growers' fields each winter was planted with highly susceptible, moderately susceptible, and partially resistant spinach inbred lines, and the plants rated weekly to calculate a Fusarium wilt severity index (FWSI) and the area under the disease progress curve (AUDPC). Results for 147 soils tested from 2010 to 2013 have been published. This study examined results for an additional 248 soils tested from 2014 to 2019 with the bioassay modified to include an option of agricultural limestone amendment to the soils tested. FWSI and AUDPC were affected significantly (p < .001) by the main effects of soil and spinach inbred line, and the interaction of these factors. Correlation analyses showed a range in degree of association of FWSI and AUDPC with spinach seed crop rotation duration and soil properties, depending on the spinach inbred line (r = −.255 to –.267, n = 172 soils with characteristics suitable for correlation analyses). Stepwise regression models for 172 soils with relevant parameters for regression analyses identified spinach seed crop rotation interval, rate of agricultural limestone amendment, soil pH, and soil Fe, Mn, and Zn concentrations as most strongly associated with FWSI and AUDPC. However, the models accounted for ≤33.4% (R²) of the variability in Fusarium wilt risk. The soil bioassay remains a primary tool for spinach seed growers to select fields with low risk of Fusarium wilt.     

Analysis of metsulfuron‐methyl residues in wheat field soil: a comparison of HPLC and bioassay techniques

BACKGROUND: Metsulfuron‐methyl is a low‐application‐rate sulfonylurea herbicide that is widely used to control broad‐leaved weeds in wheat. Owing to its persistent nature, its residues may be present at phytotoxic levels for the next crop in rotation. Therefore, a comparative evaluation of HPLC and bioassay techniques was made for the analysis of this herbicide in wheat field soil. RESULTS: Metsulfuron‐methyl was applied to wheat crop at different rates (4, 8 and 12 AI ha^?1) at 28 days after sowing as a post‐emergence application, and the soil was analysed for metsulfuron‐methyl residues by HPLC and lentil seed bioassay techniques. The bioassay was found to be the more sensitive technique. At the recommended rate of application, 4 g AI ha^?1, the bioassay technique could detect the residue up to 30 days in surface soil, while, with HPLC, residues were not detectable on the 15th day. The half‐lives of metsulfuron‐methyl by HPLC and bioassay were calculated as 6.3–7.8 and 17.5 days respectively. Under field conditions, residues of metsulfuron‐methyl were also detected in subsurface soil by the bioassay technique at trace levels, but were not detected by the solvent extraction/HPLC method. CONCLUSION: Lentil seed bioassay is a more sensitive technique than HPLC. Traces of residues detected in subsurface soil indicated the mobility of metsulfuron‐methyl into lower layers. Copyright © 2009 Society of Chemical Industry     

Optimization of hydroponic bioassay for herbicide tepraloxydim by using water free from chlorine

Initial attempts to develop a hydroponic bioassay test for tepraloxydim failed due to lack of repeatability. Investigation of the fate of tepraloxydim in test media revealed that small residues of chlorine and chloramines present on distilled water cause fast degradation of the herbicide. Half‐life of tepraloxydim in the presence of a chlorine excess was determined to be DT₅₀ < 5 s. Reaction with chloramines was slower (DT₅₀ = 4.5 h). Finally, when this factor was eliminated by using water completely free of chlorine, the main process that took place was the isomerization of the oxime group (E vs. Z). However, the overall degradation was slow (DT₅₀ = 17 days) and the hydroponic bioassay was optimized in the absence of chlorine.     

Detection of Colletotrichum coccodes and Helminthosporium solani in soils by bioassay

The sensitivity of a bioassay in detecting soil inoculum of Colletotrichum coccodes and Helminthosporium solani was examined using potato minitubers and microplants. Tests were conducted on soils which were collected from fields in which the interval after a previous potato crop differed, and which were also artificially infested with conidia or microsclerotia. For C. coccodes , determining plant infection based on the occurrence of infected roots after 9–12 weeks was a sensitive method for detecting and quantifying the amount of inoculum in soil. Infestations of less than 0·4 microsclerotia per g soil were detected in artificially infested soils. A semiselective medium, developed for isolating C. gloeosporioides from pepper, detected soil infestations by C. coccodes as low as nine conidia or one microsclerotium per g soil in artificially infested soil. For H. solani , infection on minitubers was a sensitive measure, with soil inoculum of fewer than 10 conidia per g soil being detected. Soil infestation could be quantified by assessing the percentage surface area of minitubers covered by sporulating lesions, which was strongly related to the amount of soil infestation. The results of these bioassay tests were compared with published results for real-time quantitative PCR assays on the same soils. The two methods were in good agreement in artificially infested soils, but the bioassay appeared to be more sensitive with naturally infested soils.     

土壤样品中长喙壳属真菌诱导分离方法的研究

 长喙壳属真菌(Ceratocystis spp.)是一类重要的土传植物病原真菌,可危害多种作物。由于技术方法的限制,对于土壤样品中该病原菌的分离比较困难。本研究通过比较以胡萝卜、马铃薯、甘薯作为诱导富集载体对土壤中长喙壳属真菌的富集分离效果,确定了胡萝卜为最优诱导富集载体。进一步通过对胡萝卜片厚度、土壤相对湿度、培养温度、光照条件、保湿方式等培养条件的优化,建立了从土壤中分离长喙壳属真菌的有效方法,具体为:以厚度为3~5 mm的胡萝卜片作为诱导富集载体;土壤样品相对含水量为10%~30%;12 h光暗交替或24 h黑暗;在22℃~26℃条件下不保湿进行培养,8 d后可从携带最低数量为10个孢子/g的土壤样品中分离到靶标菌。采用该方法,对采自新疆、河南、湖南、云南等地共计108份自然或耕作土壤样品进行了分离,结果表明从采自云南蒙自市、临沧市的45份土壤样品中均分离获得了长喙壳属真菌。该方法具有操作简便、所需仪器设备简单、灵敏度较高、成本低和高通量等特点,适用于田间调查,可对样本量较大的土壤样品进行快速及时分离,以获得不同土壤生境中的长喙壳属真菌分离物。     

Development of a rapid,accurate glasshouse bioassay for assessing fusarium wilt disease responses in cultivated Gossypium species

A rapid glasshouse‐based bioassay method to screen large numbers of cotton plants for responses to Fusarium oxysporum f. sp. vasinfectum (Fov) was developed. Different Fov inoculum concentrations and methods of inoculation were assessed using resistant and susceptible cotton cultivars. Cotton seeds were planted directly into Fov‐inoculated soil. Studies of seed germination, seedling establishment, seedling mortality and fusarium wilt symptoms (i.e. stunting, foliar symptoms and vascular browning) were performed to optimize the bioassay parameters. Growing seedlings in Fov‐inoculated soils at 5 × 10⁴ or 1 × 10⁵ CFU g^?1 soil, in individual seedling tubes with 12 h at 28–30°C and 12 h at 15–18°C, gave consistent results when assessing Fov disease responses 6 weeks after inoculation. When fusarium wilt resistance ranks (FWRRs) and vascular browning index (VBI) means of 18 Australian and other cotton cultivars from the Fov glasshouse bioassay were compared against their fusarium field performance ranks (F‐ranks), assessed on adult plants for cotton cultivar release, Pearson’s correlation was highly significant for both comparisons. The level of congruence between field and glasshouse data indicated that this protocol should be an effective tool for large‐scale screening for Fov‐resistance responses in diverse germplasm and breeding populations and for advancing genetic research to develop molecular markers for Fov resistance in cotton.     

一种简便易行的栗疫病菌单孢分离方法

介绍了一种分离栗疫病菌单分生孢子的简便易行的方法。此方法可以排除菌丝段的干扰,也适用于分生孢子器中产生微小分生孢子的其他真菌。