水稻内生细菌B196的鉴定及其对水稻纹枯病的防治作用

从水稻体内分离得到的细菌菌株B196对水稻纹枯病菌Rhizoctonia solani的生长有较强抑制作用.通过形态学和生理生化鉴定及16S rDNA序列分析,将菌株B196鉴定为巨大芽孢杆菌Bacillus megaterium.采用浸种和喷雾的方法将菌株B196接种到水稻植株后,均能在稻株体内回收到该菌株,表明菌株B196能在水稻体内定殖.菌株B196对水稻纹枯病的盆栽和田间防效分别为64.29%和55.13%.     

绿色荧光蛋白标记解淀粉芽孢杆菌Bam22在油菜体内的定殖

研究GFP标记的解淀粉芽孢杆菌Bam22在油菜植株上的定殖规律,为其开发利用提供科学依据.采用自然转化法将携带绿色荧光蛋白基因的质粒pGFP4412导入Bam22细胞中,构建GFP荧光标记菌株;对比野生型菌株Bam22与标记菌株Bam22-GFP的生长曲线和抑菌能力;通过灌根法,观察标记菌株在油菜体内不同组织部位的定殖...     

Macroinvertebrate frequency data for the RIVPACS III sites in Northern Ireland and some comparisons with equivalent data for Great Britain

1. A total of 313 macroinvertebrate taxa were recorded from the 70 running‐water sites in the R iver I nV ertebrate P rediction A nd C lassification S ystem (RIVPACS III) dataset for Northern Ireland, after the application of a standardization procedure to ensure that all samples were identified to the same taxonomic level. The listing includes a small number of oligochaetes not previously reported from Ireland. The frequency of occurrence of the 313 taxa within the Northern Ireland dataset is also presented. 2. The taxonomic composition and frequency of occurrence of taxa in the Northern Ireland dataset were then compared with the 614 site dataset for Great Britain, which included 637 taxa, and a 75 site subset within Britain at a similar latitude to Northern Ireland with 333 taxa. 3. The macroinvertebrate fauna of Northern Ireland is dominated by taxa recorded at the highest frequencies of occurrence in Britain. Some notable absences are highlighted and, in particular, a small number of lotic mayflies and stoneflies which are common in Great Britain. There is documentary evidence of the introduction by man of a number of non‐insect taxa, either deliberately or by accident. 4. A detailed knowledge of the present composition of the macroinvertebrate fauna of running‐water sites in Northern Ireland, and an active research programme on the potential for new colonists to pose a threat to native species, are important factors in the future conservation of the freshwater fauna. Copyright © 2000 John Wiley & Sons, Ltd.     

Migration,colonization and seedling growth of rhizobia with matrine treatment in alfalfa (Medicago sativa L.)

Purpose: The purposes of this study were to determine the effect of matrine on the migration and the colonization dynamics of the two fluorescent-tagged rhizobia in Gannong No.5 alfalfa (Medicago sativa L. Gannong No. 5) tissues, and also to determine the effect of the combination treatments on alfalfa seedlings’ growth.

Materials and methods: 0, 100, 200, 300 and 400 mg L^?1 matrine levels were added into two cyan fluorescent protein (CFP)-tagged rhizobia; Ensifer meliloti LZgn5f (gn5f) and Ensifer meliloti 12531f (12531f), respectively; and drenched the alfalfa root with the inoculants. Then the migration and colonization of the two rhizobia in alfalfa on D7, D14, D21 and D28, and subsequently seedling growth were investigated.

Results: The results showed that the optimum matrine level enhanced the colonization of both fluorescent-tagged rhizobia in alfalfa roots and the highest colonization densities of log 6.31 cfu g^?1 and log 5.87 cfu g^?1 were achieved by adding 300 mg L^?1 matrine into 12531f and adding 100 mg L^?1 matrine into gn5f, respectively. They could migrate to the aerial tissues and most colonize stems through the application of adding 300 mg L^?1 matrine into 12531f and 100 mg L^?1 matrine into gn5f, respectively. No fluorescent-tagged rhizobia were detected in the control treatment. Alfalfa seedling growth parameters like leaf chlorophyll content, seedling growth rate, root length, seedling biomass and total N percentage also increased the most when 300 mg L^?1 matrine was added into 12531f and 100 mg L^?1 matrine added into gn5f treatments.

Conclusions: Our results suggest that 300 mg L^?1 matrine added into 12531f and 100 mg L^?1 matrine added into gn5f might be exploited to promote the colonization of rhizobia in alfalfa tissue and positively impact growth and yield, indicating possible benefits for plant cultivation.     

风化煤用量下覆膜和AM真菌对玉米生长和土壤微环境的影响

为缓解我国东部贫瘠土壤,以风化煤与覆膜为切入点,通过室内盆栽试验,研究两种覆膜方式(无覆膜和覆薄膜)和两个供试土壤基质(砂土基质和砂煤混合基质)下接种AM真菌对干旱胁迫时玉米生长特性、水分利用效率与土壤性状的影响。结果表明:两个覆膜方式下,砂煤混合基质相比砂土基质提高了接种AM真菌处理的玉米根系侵染率和土壤根外菌丝密度,但无明显差异;同时土壤总球囊霉素和易提取球囊霉素含量分别显著提高了80.0%~106.5%和55.0%~73.3%(P0.05)。同一覆膜方式下,砂煤混合基质接种AM真菌和CK处理的土壤有机碳、全氮与速效磷含量分别显著高于砂土基质下相应处理,但降低了土壤速效钾含量。砂煤混合基质下覆薄膜与接种AM真菌联合对玉米株高、生物量、叶片SPAD值及水分利用效率的促进效果最好;同时砂煤混合基质接种AM真菌处理提高了无覆膜下土壤蔗糖酶和全覆膜下过氧化氢酶和碱性磷酸酶含量,分别比砂土基质下处理显著提高了46.8%~59.8%、37.9%~70.0%与57.8%~87.5%(P0.05)。研究表明施加一定量风化煤时,接种AM真菌和覆薄膜能够促进水分胁迫下的植物生长发育,改善水分利用效率和提高土壤肥力。     

华南主要树木丛枝菌根真菌物种多样性调查研究

为调查华南地区丛枝菌根（arbuscular mycorrhiza,AM）真菌的物种多样性与资源分布状况,本研究对该区域红花羊蹄甲（Bauhinia blakeana）、黄梁木（Neolamarckia cadamba）、构树（Broussonetia papyrifera）、尾叶桉（Eucalyptus urophylla）和杧果（Mangifera indica）5种主要树木AM真菌侵染状况进行研究,通过形态学特征及核糖体18S rRNA基因序列分析鉴定AM真菌种类。结果表明:1）5种树木均能形成丛枝菌根,红花羊蹄甲和尾叶桉为疆南星型（Arum-type）,杧果、构树和黄梁木为重楼型（Paris-type）。红花羊蹄甲和杧果的菌根侵染率高、孢子密度大,构树、黄梁木和尾叶桉的AM真菌侵染率和孢子密度相对较低。2）鉴定出AM真菌5属8种,分别为无梗囊霉属（Acaulospora）的浅窝无梗囊霉（Acaulospora lacunosa）和刺无梗囊霉（Acaulospora spinosa）,斗管囊霉属（Funneliformis）的摩西斗管囊霉（Funneliformis mosseae）,根孢囊霉属（Rhizophagus）的根内根孢囊霉（Rhizophagus intraradices）,以及球囊霉属（Glomus）的3种Glomus spp.和硬囊霉属（Sclerocystis）1种Sclerocystis sp.。3）球囊霉属的AM真菌广泛分布在红花羊蹄甲、杧果和构树根际,刺无梗囊霉分布在杧果和尾叶桉根际,摩西斗管囊霉为优势种,分布在构树和黄梁木根际;而浅窝无梗囊霉和硬囊霉只分布在1种树根际,表明其宿主专一性相对较强。结果显示华南树木根际土壤中AM真菌物种多样性较高,同时本研究为深入研究该地区林木根际AM真菌功能多样性提供了初步科学依据。     

Colonization of Aeromonas salmonicida subsp. masoucida strains in Atlantic salmon (Salmo salar L.) during infection

Aeromonas salmonicida subsp. masoucida (ASM) is classified as atypical A. salmonicida and brought huge economic damages to the local salmonid aquaculture in China. An ASM strain named AS‐C4 was used to investigate the colonization of ASM in Atlantic salmon (Salmo salar L.) by an immersion challenge with the control group (T0, no AS‐C4), group T1 (2.67 × 10⁴ CFU/ml AS‐C4) and group T2 (2.67 × 10⁷ CFU/ml AS‐C4). The numbers of AS‐C4 copies in different fish tissues (gill, intestine, skin, blood, muscle, spleen, liver and kidney) were determined at different time points post challenge using the quantitative real‐time PCR (qRT‐PCR). AS‐C4 were detected in the gill and intestine as early as 0 hr after the challenge both in T1 and T2 groups, suggesting that the gill and intestine were probably the portals of entry of AS‐C4 into salmon. Although AS‐C4 could not be detected in the skin until 24 hr after the challenge in T1 group, it could be detected in the skin as early as 0 hr after the challenge in T2 group, indicating that the skin may also be a portal of entry of AS‐C4 into salmon. AS‐C4 was immediately detected in the blood within 3 hr after it entered the host, suggesting that AS‐C4 successfully invaded the bloodstream of fish. After AS‐C4 colonized the host, it colonized the internal tissues, such as the spleen, liver, kidney and muscle. The results of this study will contribute to the understanding of the pathogenesis of the ASM strains and give a broader understanding of the infection route of ASM in it's host, providing more information for the development of new therapeutic strategies to protect against this pathogen in aquaculture.     

Development of a quantitative PCR assay for monitoring Streptococcus agalactiae colonization and tissue tropism in experimentally infected tilapia

Streptococcus agalactiae has become one of the most important emerging pathogens in the aquaculture industry and has resulted in large economic losses for tilapia farms in China. In this study, three pairs of specific primers were designed and tested for their specificities and sensitivities in quantitative real‐time polymerase chain reactions (qPCRs) after optimization of the annealing temperature. The primer pair IGS‐s/IGS‐a, which targets the 16S‐23S rRNA intergenic spacer region, was finally chosen, having a detection limit of 8.6 copies of S. agalactiae DNA in a 20 μL reaction mixture. Bacterial tissue tropism was demonstrated by qPCR in Oreochromis niloticus 5 days post‐injection with a virulent S. agalactiae strain. Bacterial loads were detected at the highest level in brain, followed by moderately high levels in kidney, heart, spleen, intestines, and eye. Significantly lower bacterial loads were observed in muscle, gill and liver. In addition, significantly lower bacterial loads were observed in the brain of convalescent O. niloticus 14 days post‐injection with several different S. agalactiae strains. The qPCR for the detection of S. agalactiae developed in this study provides a quantitative tool for investigating bacterial tissue tropism in infected fish, as well as for monitoring bacterial colonization in convalescent fish.     

Trichoderma asperellum is effective for biocontrol of Verticillium wilt in olive caused by the defoliating pathotype of Verticillium dahliae

Verticillium wilt caused by a highly virulent, defoliating (D) pathotype of Verticillium dahliae is threatening olive production in Spain and other Mediterranean countries. This disease must be managed by an integrated strategy, in which biocontrol agents can play an important role. We have investigated the potential of Trichoderma asperellum strains for antagonism against V. dahliae and suppression of Verticillium wilt of olive caused by the D pathotype. First, we tested the antagonistic potential of T. asperellum strains Bt2, Bt3 and T25 against six V. dahliae isolates, four of the D and two of the nondefoliating (ND) pathotypes, in different in vitro assays. All T. asperellum strains overgrew the colonies of all V. dahliae isolates to a similar extent. However, extracellular compounds from strains Bt3 and T25 showed higher anti-V. dahliae activities than those of Bt2 in membrane assays. Also, growth of Bt2 was reduced by ND V. dahliae whereas that of Bt3 and T25 was not affected by V. dahliae-secreted compounds. In planta assays using strains Bt3 and T25, and ’Picual’ olive plants, showed that the two T. asperellum strains significantly reduced the severity of symptoms and the standardized area under the disease progress curve caused by highly virulent D V. dahliae, but not the final disease incidence. Strain T25 significantly increased growth of ‘Picual’ plants and displayed higher ability for colonizing the olive rhizosphere and establishing endophytic infection in olive roots than Bt3.     

Suppression of the fungal pathogen Magnaporthe grisea by Stenotrophomonas maltophilia,a seed-borne rice (Oryza sativa L.) endophytic bacterium

The present work was carried out to study the potential of bacteria isolated from the seeds of rice plant for the biocontrol of five rice pathogenic fungi. Eleven endophytic bacteria isolated from rice seeds were evaluated for their antagonistic potential. Of five pathogens studied, only the growth of Magnaporthe grisea was inhibited by one bacterial isolate in an in vitro dual culture assay. Based on 16S rDNA sequence analysis, and biochemical and morphological characteristics, this strain was closely related to Stenotrophomonas maltophilia. We named this new isolate to be S. maltophilia SEN₁ (seed endophyte). This isolate was further tested for the production of volatile and diffusible antibiotics against M. grisea, for plant growth-promoting (PGP) traits and colonization of some rice cultivars. In addition, S. maltophilia SEN₁ was tested for its ability to promote plant growth and reduce the incidence of rice blast disease under greenhouse conditions. When applied to the soil, this isolate increased seedling growth and suppressed blast disease in plants of three studied cultivars. This study also showed this isolate could colonize the root interior of other rice cultivars. This study indicates that the S. maltophilia isolate studied has an excellent potential to be used as biocontrol agents of M. grisea or biofertilizer under in vitro and in vivo conditions.