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21.
The effects of interactions between pseudomonads (Pseudomonas cepacia strains R55 and R85, P. aeruginosa strain R80, P. fluorescens strain R92, and P. putida strain R104) and the arbuscular mycorrhizal fungus Glomus clarum (Nicol. and Schenck) isolate NT4, on spring wheat (Triticum aestivum L. cv. Laura), grown under gnotobiotic and nonsterile conditions, were investigated. Although plant growth responses varied, positive responses to pseudomonad inoculants generally were obtained under gnotobiotic conditions. Shoot dry weight enhancement ranged from 16 to 48%, whereas root enhancement ranged from 82 to 137%. Shoot growth in nonsterile soil, however, was unaffected by pseudomonad inoculants, or reduced by as much as 24%. Shoot growth was unaffected or depressed by G. clarum NT4 whereas early root growth was enhanced by 38%. Significant interactions between the pseudomonad inoculants and G. clarum NT4 were detected. Typically, dual inoculation influenced the magnitude of response associated with any organism applied alone. The effect of these pseudomonads on G. clarum NT4 spore germination was investigated. Germination was inhibited when spores were incubated either on membranes placed directly on bacterial lawns of strains R85 and R104 (i.e., direct assay), or on agarose blocks separated from the bacteria by membranes (i.e., diffusion assay). When the agarose blocks were physically separated from the pseudomonad (i.e., volatile assay), there was no evidence of inhibition, suggesting that a nonvolatile, diffusible substance(s) produced by both strains R85 and R104 may inhibit G. clarum NT4 spore germination. Received: 11 December 1995  相似文献   
22.
Dried soil samples from many sources have been stored in archives world-wide over the years, but there has been little research on their value for studying microbial populations. Samples collected since 1843 from the Broadbalk field experiment on crop nutrition at Rothamsted have been used to document changes in the structure and composition of soils as agricultural practices evolve, also offering an invaluable record of environmental changes from the pre- to post-industrial era in the UK. To date, the microbial communities of these soils have not been studied, in part due to the well-documented drop in bacterial culturability in dried soils. However, modern molecular methods based on PCR amplification of DNA extracted directly from soil do not require bacterial cells to be viable or intact and may allow investigations into the legacy of bacteria that were present at the time of sample collection.

In a preliminary study, to establish if dried soils can provide a historical record of bacterial communities, samples from the Broadbalk soil archive dating back to 1868 were investigated and plots treated with either farmyard manure (FYM) or inorganic fertilizer (NPK) were compared. As anticipated, the processes of air-drying and milling greatly reduced bacterial viability whilst DNA yields declined less and may be preserved by desiccation. A higher proportion of culturable bacteria survived the archiving process in the FYM soil, possibly protected by the increased soil organic matter. The majority of surviving bacteria were firmicutes, whether collected in 2003 or in 1914, but a wide range of genera was detected in DNA extracted from the samples using PCR and DGGE of 16S rRNA genes. Analysis of DGGE band profiles indicated that the two plots maintained divergent populations. Sequence analysis of bands excised from DGGE gels, from a sample collected in 1914, revealed DNA from - and β-proteobacteria as well as firmicutes. PCR using primers specific for ammonia oxidizing bacteria showed similar band profiles across the two treatments in recently collected samples, however older samples from the NPK plot showed greater divergence. Primers specific for the genus Pseudomonas were designed and used in real-time quantitative PCR to indicate that archived soil collected in 1868 contained 10-fold less pseudomonad DNA than fresh soil, representing around 105 genomes g−1 soil. Prior to milling, dramatically less pseudomonad DNA was extracted from recently collected air-dried soil from the NPK compared to the FYM plot; otherwise, the two plots followed similar trends. Overall bacterial abundance, diversity and survival during the archiving process differed in the two soils, possibly due to differences in clay and soil organic matter content. Nevertheless, the results demonstrate that air-dried soils can protect microbial DNA for more than 150 years and offer an invaluable resource for future research.  相似文献   

23.
弹性蛋白酶基因(PAE)的克隆及在毕赤酵母中的表达   总被引:1,自引:0,他引:1  
以1株产弹性蛋白酶的铜绿假单胞菌(Pseudomonas aeruginosa)基因组DNA为模板,经PCR扩增得到的铜绿假单胞菌弹性蛋白酶(P.acruginosa elastase,PAE)基因,与GenBank中的序列对比发现同源性为99%.成功地构建了重组表达载体pPIC3.5K/PAE,莺组质粒Sac Ⅰ线性化后转化毕赤酵母(Pichia pastoris)菌株KM71中,通过PCR和表型鉴定表明,PAE基因已经整合到毕赤酵母染色体上.经大量筛选获得48株含高拷贝的重组毕赤酵母转化子.在甲醇诱导下,经过毕赤酵母高密度发酵进行PAE的表达,经SDS-PAGE分析.结果表明,在培养基上清中含有一明显特异性蛋白条带,大小为34kD.活性检测结果,酶活为1 060 U/mL,是出发菌株的26倍.  相似文献   
24.
基于全基因组数据,确定生防菌株PF-1的分类地位,验证其对植物病原菌的拮抗作用以及挖掘其潜在的生防功能。通过全基因组中16S rRNA基因序列的系统发育分析及基因组分析方法确定生防菌株PF-1的分类地位,通过平板对峙方法研究其对马铃薯枯萎病菌和棉花黄萎病菌的拮抗能力;利用antiSMASH软件分析和预测菌株PF-1的抗生素相关基因并挖掘其生防潜力。基于16S rRNA基因序列的系统发育分析及基因组分析结果,PF-1被鉴定为防御假单胞菌(Pseudomonas protegens),同时发现PF-1基因组中存在15种次生代谢产物基因簇,具有良好的抑制病原菌生长和提高植物抗病性的能力,在农业中具有良好的应用前景。  相似文献   
25.
番茄青枯菌分泌物中热稳定成分的毒性研究   总被引:5,自引:0,他引:5  
研究了青枯细菌毒性菌株平板培养滤液中热稳定性成分对番茄组织培养物的影响。结果表明;PCF中含有一类非酶,非胞外多糖的热稳定毒性物质,它对番茄叶片脱分化过程有抑制作用,抗病材料较感病材料敏感,另外,它对番茄试管生根也起抑制作用;EPS没有上述抑制作用。  相似文献   
26.
2000-2001年在江苏、浙江、安徽等地发现一种桔梗病害,并从其病叶上分离得到了26个菌株。菌株接种于桔梗叶片后,发病症状与自然发病症状完全一致,并从回接病株上重新分离得到此病原菌。各菌株间致病力无明显的差异。经革兰氏染色反应、菌体形态、培养性状、生理生化反应、(G C)%等鉴定,确定该病原菌为丁香假单胞杆菌的一个新的致病变种。该病菌能引起桔梗细菌性叶斑病(又称斑点病)。  相似文献   
27.
定期测定了感病品种红花大金元接种烟草野火病菌后叶片内5种酶活性的动态变化,研究结果表明:烟草接种病菌后,SOD活性先上升,后在8d下降,低于对照;POD活性接种后在1d略低于对照,后上升较快,10d达到高峰,此后一直高于对照;PPO活性在接种后1d低于对照15.8%,但此后上升,16d达到高峰,18d下降低于对照;CAT活性变化与POD相似,接种1d低于对照,但此后一直高于对照,并于6d达到高峰,10d虽有所下降,但接着升高;PAL活性与CAT、POD变化相似,接种后1d活性低于对照28.3%,其后上升,10d达到高峰,是对照的2.11倍,并维持较高的水平。  相似文献   
28.
姜瘟病发生规律调查及防治建议   总被引:2,自引:1,他引:2  
通过对舒城、铜陵、临泉、体宁等主要栽姜区姜瘟病发生规律的调查、病原菌鉴定和防治研究,明确了引起姜瘟病的病原菌为姜青枯假单胞菌,除少数菌株属生物型Ⅲ外都为生物型Ⅳ。新栽姜区的菌原主要来自带病种姜,而老姜区土壤带菌和带病种姜都是主要侵染源。病害发生与轮作、种姜处理、气候、姜田管理、土质、施肥等关系密切。采取轮作、选留无病种姜建立留种田、种姜处理、加强田间管理等,结合大田期药物防治能有效的控制病害。  相似文献   
29.
对杨树高效解磷细菌蜡状芽孢杆菌(Bacillus cereus)JYZ-SD1和荧光假单胞菌(Pseudomonas fluorescens)JW-JS1在3L发酵罐里发酵条件进行优化。结果显示这两株解磷细菌在72h内都是接种量20%、转速400 r/min、通气量100ln/h条件下菌体生长量最大。为将这两株杨树根际解磷细菌更高效、更大量的扩繁提供参考依据。  相似文献   
30.
从全国各地采集豆豉样品,经富集培养并利用纤维蛋白平板法获得一株形态与现存产纤溶酶微生物差异较大的菌株HS9。通过传统方法、化学方法以及16SrRNA序列分析对HS9进行分类鉴定,属于Pseudomonas aeruqinosa,是未见报道的产豆豉纤溶酶菌株。发酵培养HS9获得粗酶,经20%~70%硫酸铵梯度盐析、SephadexG-75凝胶过滤以及CM-Sepharose Fast Flow阳离子交换层析分离纯化后,得到了电泳纯酶。通过SDS-PAGE了解该酶分子量约为34kD,pH8.0~8.5时酶活性最高,最适作用温度48℃,作用方式为直接水解纤维蛋白,胃蛋白酶抑制剂在工作浓度1μmol/L时能完全抑制其活性,推测该酶为天冬氨酸蛋白酶,是一种新型的豆豉纤溶酶。  相似文献   
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