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1.
Previous studies showed that Burkholderia pyrrocinia JK‐SH007 is a potential biocontrol agent of poplar canker disease. In this report, B. pyrrocinia JK‐SH007 was genetically modified by introducing the Bacillus subtilis Chi113 gene to enhance its antifungal activity. The green fluorescence of B. pyrrocinia JK‐SH007E1 can be detected because of its transformation with plasmid pHKT2‐Chi113 containing a gfp reporter gene. Real‐time quantitative PCR (qPCR) showed that the B. subtilis Chi113 gene was highly expressed in B. pyrrocinia JK‐SH007E1 at mRNA level. The chitinase activity and the antifungal activity of B. pyrrocinia JK‐SH007E1 were significantly increased, and the clear halo was visible on the colloidal chitin agar plate. In addition, the constructed recombinant plasmid pHKT2‐Chi113 was stably maintained for at least 100 generations in the absence of antibiotic selection in B. pyrrocinia JK‐SH007E1. In western blot analysis, the Chi113‐GFP fusion protein was detected in B. pyrrocinia JK‐SH007E1 using the GFP polyclonal antibody. In conclusion, the B. subtilis Chi113 gene was successfully transformed into B. pyrrocinia JK‐SH007 and was expressed both at mRNA level and protein level. Moreover, the antifungal activity of B. pyrrocinia JK‐SH007 was improved through this genetic modification.  相似文献   

2.
【目的】探究荧光假单胞菌与红绒盖牛肝菌共接种条件下NL-895杨根际土壤酶活性和土壤微生物功能多样性的变化,从微生态水平揭示溶磷细菌与外生菌根真菌互作对NL-895杨的促生机制。【方法】采用盆栽试验考察荧光假单胞菌JW-JS1与红绒盖牛肝菌(Xc)共接种后NL-895杨根际土壤酶(酸性磷酸酶、脱氢酶和转化酶)的活性变化,并通过BIOLOG微孔板法分析共接种对NL-895杨根际土壤微生物群落和功能多样性的影响。【结果】从整体趋势来看,不同接种处理后NL-895杨根际土壤酶活性均有所增强,土壤酸性磷酸酶、脱氢酶和转化酶活性均大于对照,其中,共接种JW-JS1+Xc处理对土壤酸性磷酸酶和转化酶活性的促进最为明显;接种150天后,不同接种处理土壤微生物的AWCD值均高于CK(培养24~48 h除外),且随着培养时间的延长呈先上升后稳定的趋势;培养96 h后,共接种JW-JS1+Xc处理土壤微生物McIntosh优势度指数和碳源利用丰富度指数显著高于单接种Xc或JW-JS1和CK(P<0.05),而Shannon丰富度指数和Simpson优势度指数显著低于单接种Xc或JW-JS1和CK(P<0.05);不同接种处理NL-895杨根际土壤微生物对BIOLOG微孔板中6大类碳源的利用能力差异显著(P<0.05)。【结论】荧光假单胞菌JW-JS1与外生菌根真菌Xc共接种对NL-895杨根际土壤酶活性和土壤微生物功能多样性的提高具有一定的促进作用,有利于其土壤肥力的改善,进而促进NL-895杨的生长。与单接种相比,溶磷细菌与外生菌根真菌共接种可更有效地提高杨树根际土壤酶活性和土壤微生物多样性。  相似文献   

3.
采集江西省林科院竹种园刚竹属3个竹种——毛竹、雷竹及黄秆乌哺鸡竹根际土壤作为研究对象,采用高通量测序技术比较3个竹种根际土壤微生物群落结构,分析其种类及多样性的差异。结果表明:不同竹种根际微生物群落结构具有一定差异性。3个竹种根际土壤细菌群落在各级分类水平上组分相似,构成比例差异不大。真菌群落在各级分类水平上组分相似,构成比例差异显著。在3个竹种中毛竹和雷竹根际土壤固氮菌群落构成组分及比例都更接近。由此可见,不同竹种的根际微生物群落都有其特异性,因此在培育中应对不同的竹种选择施用适当的微生物菌肥,以促进竹种根际土壤优势菌群的形成,进而促进竹种的生长。  相似文献   

4.
The fungal pathogen Septoria musiva can be difficult to isolate from cankers that result from its colonization of poplar stems, and its persistence in these cankers has not been well studied. In order to compare cultural and polymerase chain reaction (PCR)‐based assays for detection of S. musiva in cankers, stems of susceptible hybrid poplar clone NC11505 were wounded and inoculated in August 2003. At 8, 16, 24 and 32 weeks after inoculation (October and December 2003, February and April 2004, respectively), 110 inoculated stems (plus controls) were harvested and a semiselective culture medium was used in attempts to detect the pathogen in bark and wood. Six chips of bark and six chips of underlying wood from one half of each canker were incubated on the semiselective medium for 2 weeks until pycnidia and conidia of S. musiva could be identified. The number of positive cankers and positive chips (out of six attempts per tissue per canker) was recorded. The remaining halves of cankers from subsets of 70 inoculated stems (plus controls) of those harvested in October 2003 and April 2004 were tested using a PCR‐based assay. Three chips of bark and three chips of underlying wood were ground, and DNA was extracted and then amplified using S. musiva‐specific primers designed from the internal transcribed spacer (ITS) region of nuclear rDNA repeats. The number of positive cankers and positive chips (out of three attempts per tissue per canker) was recorded. For both assays, the number of positive cankers and the number of positive chips per canker decreased with time. Using either assay, however, the pathogen was still detected from at least 49% of cankers at 32 weeks after inoculation.  相似文献   

5.
A series of experiments was conducted to determine the relative tolerance in vitro of an isolate of Septoria musiva (a fungus that causes a severely damaging stem canker disease of poplars) for selected chemicals. Inhibition of diameter growth of this fungus on a V‐8 vegetable juice‐based medium with captan, chlorothalonil, iprodione, mancozeb and streptomycin sulphate at concentrations, respectively, of 50, 1, 10, 10, and 100 mg l–1 was relatively low compared to inhibition of eight other fungi cultured from cankers on poplars. In addition, the presence of captan stimulated profuse sporulation of the fungus. These properties assisted in the identification of S. musiva from cankers resulting from artificial inoculation of poplar branches in the field.  相似文献   

6.
A study of lesion development in stems of Eucalyptus nitens following artificial inoculations with canker fungi was carried out on 16‐year‐old plantation trees. In a first trial cambium bark wounds on smooth‐ and rough‐barked trees were inoculated with the mycelium of nine species of canker fungi, including Endothia gyrosa. In a second trial spores or mycelium of E. gyrosa were applied directly onto undamaged or superficially wounded bark surfaces. Infection subsequent to artificial inoculation via wounding (whatever the wounding technique or type of inoculum) resulted in significantly larger external lesions (mean lesion area up to 35.6 cm2 20 months after inoculation) on smooth bark compared with those on rough bark (up to 19.0 cm2). Microscopic studies of infected rough and smooth bark suggest that, once smooth bark is compromised by wounding and artificial inoculation, the particular anatomical structure of smooth bark may offer less mechanical resistance to post‐penetration hyphal spread in comparison with rough bark. It is suggested that at a pre‐penetration stage under natural conditions spores of E. gyrosa more easily infect rough bark via cracks associated with this type of bark but not present in smooth bark.  相似文献   

7.
Intercropping of mulberry(Morus alba L.)and alfalfa(Medicago sativa L.) is a new forestry-grass compound model in China,which can provide high forage yields with high protein.Nitrogen application is one of the important factors determining the production and quality of this system.To elucidate the advantages of intercropping and nitrogen application,we analyzed the changes of physicochemical properties,enzyme activities,and microbial communities in the rhizosphere soil.We used principal components analysis(PCA) and redundancy discriminators analysis to clarify the relationships among treatments and between treatments and environmental factors,respectively.The results showed that nitrogen application significantly increased pH value,available nitrogen content,soil water content(SWC),and urea(URE) activity in rhizosphere soil of monoculture mulberry.In contrast,intercropping and intercropping+N significantly decreased pH and SWC in mulberry treatments.Nitrogen,intercropping and intercropping+N sharply reduced soil organic matter content and SWC in alfalfa treatments.Nitrogen,intercropping,and intercropping+N increased the values of McIntosh diversity(U),Simpson diversity(D),and Shannon-Weaver diversity(H') in mulberry treatments.However,PC A scatter plots showed clustering of monoculture mulberry with nitrogen(MNE) and intercropping mulberry without nitrogen(M0).Intercropping reduced both H' and D but nitrogen application showed no effect on diversity of microbial communities in alfalfa.There were obvious differences in using the six types of carbon sources between mulberry and alfalfa treatments.Nitrogen and intercropping increased the numbers of sole carbon substrate in mulberry treatments where the relative use rate exceeded 4%.While the numbers declined in alfalfa with nitrogen and intercropping.RDA indicated that URE was positive when intercropping mulberry was treated with nitrogen,but was negative in monoculture alfalfa treated with nitrogen.Soil pH and SWC were positive with mulberry treatments but were negative with alfalfa treatments.Intercropping with alfalfa benefited mulberry in the absence of nitrogen application.Intercropping with alfalfa and nitrogen application could improve the microbial community function and diversity in rhizosphere soil of mulberry.The microbial community in rhizosphere soil of mulberry and alfalfa is strategically complementary in terms of using carbon sources.  相似文献   

8.
[目的]通过分析土壤微生物生物量及群落结构的演变趋势,筛选影响土壤微生物群落的关键环境因子,揭示土壤微生物群落对毛竹林长期粗放经营的响应机理。[方法]选取不同粗放经营年限(5 a、9 a、15 a、18 a)毛竹林,以天然马尾松林(Masson pine,MP)作为对照,采用磷脂脂肪酸(Phospholipid fatty acids,PLFA)分析方法表征土壤微生物量及群落结构。[结果]结果表明,毛竹林土壤微生物总PLFA含量以及细菌、真菌、放线菌等PLFA含量均显著低于马尾松林(P0.05),但不同经营年限毛竹林之间没有显著差异。土壤碱解氮及有效磷含量对土壤微生物总PLFA含量以及细菌、真菌、原生动物等PLFA含量影响显著(P0.05),而土壤碱解氮、p H值以及有机质含量对放线菌PLFA含量影响显著(P0.05)。长期粗放经营过程中毛竹林土壤微生物丰富度及多样性均呈逐渐下降趋势。非度量多维尺度转换排序(Non-metric multidimensional scaling,NMDS)分析结果表明,毛竹林土壤微生物群落结构与马尾松林有明显区分(R=0.388 1,P=0.009)。土壤含水量、碱解氮、有效磷以及p H值合计解释了90.28%的微生物群落结构变异量,其中土壤含水量、碱解氮、有效磷贡献显著(P0.05)。[结论]长期粗放经营降低了毛竹林土壤微生物量,改变了群落结构,其生态风险还有待于进一步评估。  相似文献   

9.
Phytophthora cinnamomi was repeatedly isolated from the rhizosphere of recently dead sweet chestnut trees in the upper Rhine valley. Identification was genetically confirmed by RFLP and sequencing of the ITS‐region. This is the first record of P. cinnamomi on forest land in Germany. Mode of introduction of this invasive species and potential factors favouring establishment are discussed.  相似文献   

10.
This study describes the first observation of Botryodiplodia canker in the Western Carpathians in south‐eastern Poland caused by Botryodiplodia hypodermia (Sacc.) Petr. (syn. Sphaeropsis hypodermia, S. ulmicola). The canker occurred on an approximately 17‐year‐old Ulmus glabra sapling in a mixed conifer/deciduous stand with elm trees severely damaged by Dutch elm disease. This paper describes disease symptoms and provides information on the macro‐ and micromorphology of the fungus isolated from the cankered tissues. The results of BLAST search using DNA sequences obtained for our cultures and subsequent phylogenetic positioning of the fungus among closely related Botryosphaeriaceae indicate that the species is much more closely related to Phaeobotryon than to the other Botryodiplodia or Sphaeropsis species. Moreover, a total of 16 polymorphisms within the ITS region were detected between S. ulmicola associated with Botryodiplodia canker in North America and B. hypodermia associated with the canker observed in Poland. Thus, the “European” variant of “Sphaeropsisulmicola can now be easily identified with our barcode sequences. The Botryodiplodia canker is much less prevalent in Europe than in North America. Differences in virulence of “American” and “European” linages and differences in susceptibility of various elm species may be the reason for the higher prevalence of the disease in North America.  相似文献   

11.
Europeanalder seedlings were inoculated with a suspension of the putative plant growthpromoting rhizobacterium (PGPR) Bacillus pumilus (CECT5105), or left non-inoculated (controls) in two different soils, and grownundercontrolled conditions. Soil A showed a thick texture, slightly acidic with ahigh mineral nitrogen content, while soil B showed a thin texture, basic andwith a lower nitrogen content. At each sampling time, over an 8-week period,shoot and root systems of the plants were measured, nodules counted, and shootand root length and surface were determined. In addition, changes in themicrobial rhizosphere structure were evaluated by the phospholipid fatty acid(PLFA) profile extracted directly from the rhizosphere soil. The increasesdetected in shoot surface were significant only in soil A, while the rootsystemwas affected in both soils. In soil A, inoculation with B.pumilus caused a perturbation that subsequently disappeared, whilethe rhizosphere community structure was seriously altered in soil B. Allbiometric parameters were enhanced to a greater extent in soil A, in which theinoculum did not alter the existing rhizosphere communities and nutrientavailability was better.  相似文献   

12.
In a 4‐year‐old collection of native Italian Populus alba, growing in the eastern Po valley, many trunks showed severe symptoms characterized by brown spots, similar to trunk scab and/or by cankers. Canker‐like growths, often with gall‐like‐formations in the earlier developmental phases were also found on some clones of P. trichocarpa and their hybrids, as well as on P. deltoides growing in several Northern Italian nurseries. Bacteria and fungi present in the affected tissues and potentially involved in the disease were examined. No fungi were detected, however, several species of bacteria were isolated and identified: Pseudomonas mendocina and Erwinia herbicola group, from the brown spots; Erwinia carotovora sub sp. carotovora and, occasionally, E. herbicola group, from cankers. Only E. carotovora sub sp. carotovora produced clear symptoms of canker when artificially inoculated on young plants of different poplar species. This is the first time this symptomology on poplar species has been clearly related to E. carotovora sub sp. carotovora.  相似文献   

13.
以长江中游芦苇滩地为研究对象,利用微生物分离培养计数方法和PCR-DGGE技术重点研究了人工杨树林对芦苇滩地土壤细菌种群的影响。结果表明:与芦苇地相比,人工杨树林根际土、根表土和0-10cm土层中的氮、速效磷和总有机碳的含量都明显降低;人工杨树林下根际土中的微生物数量都比芦苇地的低,而其根表土中则是细菌、真菌数量低于芦苇地,放线具比芦苇地高;就4种功能细菌来说,人工杨树林根际土的纤维素菌、反硝化菌、硝化细菌和氨氧化细菌的数量则比芦苇地低,而根表土中除了氨氧化细菌外,其他3种功能细菌含量又比芦苇地的高。4种功能细菌数量在不同的土壤剖面中的分布也各不相同:在0-10cm土层中,人工杨树林的纤维素菌和氨氧化细菌的数量高于芦苇地,而其余2种功能茵则低于芦苇地;在10-25cm土层中,人工杨树林的纤维素菌和硝化细菌含量低于芦苇地,反硝化菌的含量却高于芦苇地,而氨氧化菌含量相同;在25-60cm土层中,除了纤维素菌,其他3种功能细菌含量都是人工杨树林低于芦苇地。通过系统进化树分析发现,滩地中普遍存在的细菌类群为α变形菌纲、β变形茵纲和硝化螺旋菌纲,而人工杨树林和芦苇地的根表土、根际土和0-10cm土层均存在各自专有的细菌种群,表明人工杨树林对芦苇滩地土壤中细菌种群产生了一定的影响。  相似文献   

14.
除草剂草甘膦对板栗根际土壤微生物多样性的影响   总被引:3,自引:0,他引:3  
近年来,除草剂的使用已成为提高作物产量和品质的一种常用手段,伴随而来的是土壤微生物多样性失调及污染严重等生态问题。为了促使板栗能健康生长并保证其果实品质,利用Illumina Miseq高通量技术,分别对草甘膦施用后不同时间采集的根际土壤样品中细菌16S rRNA的V3-V4区和真菌ITS1-ITS2区进行测序,分析微生物群落的组成和多样性的变化情况。多样性分析结果表明:以草甘膦处理6和12d后,板栗根际土壤细菌的多样性和丰富度均无显著变化,真菌多样性显著降低,真菌丰富度先增加后不变。群落结构分析结果表明:板栗根际土壤优势细菌菌门主要是变形菌门、放线菌门、酸杆菌门等,优势真菌菌门主要是子囊菌门和担子菌,随着草甘膦处理时间的增加,变形菌门、子囊菌门的相对丰度均先增加后不变,酸杆菌门的相对丰度先增加后降低,担子菌门的相对丰度先增加后保持不变。研究结果表明:草甘膦对板栗根际土壤微生物多样性及群落组成均产生了一定的影响,其影响随着施药时间的延长而发生变化。  相似文献   

15.
Corymbia calophylla (marri), a keystone tree species in the global biodiversity hot spot of southwestern Australia, is suffering decline and mortality associated with a canker disease caused by the endemic fungus Quambalaria coyrecup. Phytophthora species are frequently isolated from the rhizosphere of C. calophylla, and a hypothesis is that Phytophthora root infection is predisposing C. calophylla to this endemic canker pathogen. Field surveys were conducted in both anthropogenically disturbed and undisturbed C. calophylla stands, from where a total of 100 rhizosphere soil samples, from both healthy and cankered trees, were collected. Phytophthora species were isolated from 26% of the samples collected, with Phytophthora incidence significantly higher on disturbed stands than in natural forests (73% and 27%, respectively). Five Phytophthora species were recovered, including P. cinnamomi, P. elongata, P. multivora, P. pseudocryptogea and P. versiformis. Under‐bark inoculations with the Phytophthora isolates caused significant lesion lengths in excised C. calophylla stems. Corymbia calophylla response to pot infestation trials in the glasshouse varied between Phytophthora species and isolates, with isolates of P. cinnamomi and P. multivora causing a significant reduction in seedling root volume and often leading to seedling death. This study demonstrates that root disease caused by Phytophthora species, especially P. cinnamomi and P. multivora, has the ability to adversely affect C. calophylla health. This study leads the way to do a dual inoculation trial with the canker pathogen Q. coyrecup, and different Phytophthora species to investigate if Phytophthora root infection predisposes C. calophylla to this canker disease.  相似文献   

16.
To improve the productivity of poplar plantations, a field experiment of split-plot design with four tree spacings and three poplar clones was established, and four soil enzyme activities and microbial biomass were monitored in the trial.Soil enzyme activities, in most cases,were significantly higher in topsoil(0–10 cm) than in lower horizons(10–20 cm).Soil cellulase, catalase and protease activities during the growing season were higher than during the non-growing season, while invertase activity followed the opposite trend.Soil invertase, cellulase and catalase activities varied by poplar clone but soil protease activity did not.Cellulase and protease activities in the plantation at 5×5 m spacing were significantly higher than in the other spacings.The highest catalase activity was recorded at 6×6 m spacing.At the same planting density, invertase activity was greater in square spacings than in rectangular spacings.Soil microbial biomass was also significantly affected by seedling spacing and poplar clone.The mean soil MBC was significantly lower in topsoil than in the lower horizon, while MBN showed the opposite pattern.Significantly positive correlations were observed among soil cellulase, protease and catalase activities(p0.01), whereas soil invertase activity was negatively and significantly correlated with cellulase, protease and catalase activities(p0.01).Soil microbial biomass and enzyme activities were not correlated except for a significantly negative correlation between soil MBC and catalase activities.Variations in soil enzyme activity and microbial biomass in different poplar plantations suggest that genotype and planting spacing should be considered when modeling soil nutrient dynamics and managing for long-term site productivity.  相似文献   

17.
The isolation, culture and the active determination of poplar ice nucleation active (INA) bacteria and the inoculation tests in laboratory and field were conducted, and the varieties, distribution and number of poplar INA bacteria and its pathogenicity and freezing injury property were determined. The study results showed that the INA bacteria widely spread on poplar in Northeast China and caused the frozen injury for poplar under the frost condition in Spring or Autumn, which was the key factor to induce INA bacterial canker. Through evaluation and investigation of different poplar varieties and inoculation tests, fine disease-resistant varieties and strains of poplar suitable for Northeast China were selected. Further tests for strong seedling showed that burying cuttings in sand and covering with plastic film could effectively avoid the frostbite, frozen and drought damage, reduce INA bacteria infection, and promote poplar growth. INA bacterial canker was detected early by highly specialized antiserums of INA bacteria and the agglutinated test of ring-shaped boundary surface. The inducers such as streptomycin, phenylmercuric acetae, salicylic acid and heat-killed bacteria to immerse cuttings, have obvious induced disease-resistant effect. Before poplar sprouted in early spring, through spraying the solution of frostbite agent, the control effect also was obvious. Foundation item: This paper was supported by National Foundation of Ninth Five-Year Plan (No.96-005-04-01-03). Biography: XIANG Cun-ti (1933-), male, professor in Collage of Forest Resources and Environment of Northeast Forestry University, Harbin 150040, P.R. China. Responsible editor: Zhu Hong  相似文献   

18.
麻疯树是公认的生物燃料植物之一,也是一种具有修复土壤重金属污染潜力的物种。近年来,基于麻疯树速生和各器官含有毒物质,麻疯树对土壤产生的影响越来越受到关注。文中综述了麻疯树对土壤物理性状、化学性状、微生物数量、微生物群落组成及土壤酶活性变化的影响,展望了未来的研究方向。研究表明,在不同种植条件下麻疯树引起土壤水分含量、pH值、主要养分含量及微生物多样性变化的趋势不同,麻疯树对土壤性状影响的研究存在一定局限性。建议深入研究麻疯树种植及其凋落物对土壤性状的影响,以期为麻疯树种植与调控提供科学参考。  相似文献   

19.
Abstract

Selected microorganisms (Laccaria laccata, Trichoderma viride, Bacillus subtilis, Pseudomonas corrugata and Azotobacter chroococcum) were used as inoculants to study their influence on seed germination, seedling survival and subsequent growth in Cedrus deodara. The microbial inoculations positively affected these parameters over control, in the following order: Azotobacter chroococcum Pseudomonas corrugata Bacillus subtilis Trichoderma viride Laccaria laccata.The major cause of seedling mortality was found to be (1) wilting of emerged seedlings by Fusarium oxysporum, and (2) cutworm attack. Bacterial inoculations were found to be superior in suppressing the proliferation of Fusariumsp., thereby enhancing seedling survival. One of the bacterial inoculants, B. subtilis, resulted in 76% seed germination, compared to 54% in control. Though inoculation with L. laccataresulted in maximum mycorrhizal infection (up to 72%) compared to48%in control seedlings, it afforded least protection against Fusariumsp., as well as minimal growth promotion. On the other hand, inoculation with T. viridesignificantly controlled wilting of seedlings and improved plant growth. Microbial inoculations were found to influence the soil microflora, nutrient status of the rhizosphere soil and that of different parts of seedlings. Enrichment of carbon was recorded in the rhizosphere soil. Enhanced nitrogen, phosphorus and potassium content of various plant parts indicated a positive influence also on nutrient uptake. The present study has implications for large scale forestry programs.  相似文献   

20.
Microbe communities in rhizosphere ecosystems are important for plant health but there is limited knowledge of them in the rhizospheres of genetically modified(GM) plants, especial for tree species. We used the amplitude sequencing method to analyze the V4 regions of the 16 S r RNA gene to identify changes in bacterial diversity and community structure in two GM lines(D520 and D521), one non-genetically modified(nonGM) line and in uncultivated soil. After chimera filtering,468.133 sequences in the domain Bacteria remained. There were ten dominant taxonomic groups(with [1 % of all sequences) across the samples. 241 of 551 genera(representing a ratio of 97.33 %) were common to all samples.A Venn diagram showed that 1.926 operational taxonomic units(OTUs) were shared by all samples. We found a specific change, a reduction in Chloroflexi, in the microorganisms in the rhizosphere soil planted with poplars. Taken together, the results showed few statistical differences in the bacterial diversity and community structure between the GM line and non-GM line, this suggests that there was no or very limited impact of this genetic modification on the bacterial communities in the rhizosphere.  相似文献   

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