生物炭对设施黄瓜根际土壤养分和菌群的影响

以设施黄瓜根际土壤为试验对象,通过每hm2添加5、10、20、40、60 t不同生物炭量作为处理,研究黄瓜结果期养分和细菌群落变化规律,探讨生物炭调控二者间相互作用机制。试验结果表明:在生物炭添加量为5~60 t/hm2时,在黄瓜结果期可不同程度提高根际土壤碱解氮、速效磷、速效钾、有机碳含量和全氮质量比;在20~60 t/hm2时,可明显提高结果盛期根际土壤细菌中变形菌门、放线菌门、厚壁菌门丰度而降低酸杆菌门、绿弯菌门丰度;生物炭通过调控并提高变形菌门γ-变形菌纲、放线菌门放线菌纲放线菌目等细菌数量和比例,促进根际土壤碱解氮、速效磷、速效钾质量比的提高;经综合比较,20 t/hm2的生物炭量处理效果优于其他处理,与对照相比,在结果盛期可提高根际土壤碱解氮质量比39.86%、速效磷质量比135.95%、速效钾质量比81.35%、有机碳质量比82.89%、全氮质量比73.77%。     

炭化稻壳复合土壤调理剂改善土壤环境提高山药产量

山药的生长发育与土壤环境质量关系密切,因此,土壤质量的改善是提高山药抗性与产量的必要措施。本研究在江西瑞昌山药种植区通过施用土壤调理剂对山药土壤进行处理,旨在探明土壤调理剂对山药土壤理化性质和细菌微生物群落组成的影响,为山药土壤微生态环境的改良和栽培管理措施提供有效的技术支持。结果表明:1)土壤调理剂改变了土壤的理化性质,调理剂处理的土壤p H值、有机质含量、全氮含量、有效磷含量、速效钾含量、土壤含水量、团聚体几何平均直径、总孔隙度均增加,而土壤容重下降;2)与对照相比,调理剂处理的土壤中细菌多样性较丰富,其中酸杆菌门的相对丰度明显增加,变形菌门的相对丰度下降,GAL15和AD-3门的相对丰度明显下降,绿弯菌门、疣微菌门、芽单孢菌门的相对丰度明显增加,浮霉菌门的相对丰度明显下降,拟杆菌门的相对丰度明显增加;3)土壤调理剂增加了山药生长期叶片中叶绿素和脯氨酸的累积,提高了山药块茎组织超氧化物歧化酶的活性,提升了山药块茎抗病虫害能力及产量。研究表明土壤调理剂可通过改善土壤的理化性质及土壤中某些细菌的相对丰度,显著改善山药土壤的微生态环境,提高山药抗病虫害侵染能力,为山药品质改善及产量增加提供了有效的手段。     

Rhizobium strain,a banana (Musa spp.)-associated bacterium with a high potential as biofertilizer

A diazotrophic bacterial strain denominated 11B isolated from the rhizosphere of a banana plant (Musa spp.) was characterized morphologically, biochemically, and phylogenetically, whereas the symbiotic potential of the strain was assessed through tests of host range, interstrain nodulation competitiveness, and capacity to synthesize indole-3-acetic acid (IAA) and solubilize phosphate. Based on morphological and physiological–biochemical properties, as well as 16S ribosomal deoxyribonucleic acid (rDNA) sequence and phylogenetic analysis, the strain 11B belonged to the genus Rhizobium with 100.0% sequence similarity with Rhizobium tropici CAF440. The optimum growth temperature and pH for strain 11B are 28°C and 7.2, respectively. This strain was able to produce IAA, solubilize phosphate, and fix large amounts of nitrogen (N₂) and form effective nodules on the legumes Acaciella angustissima, Gliricidia sepium, Leucaena leucocephala, Lysiloma acapulcensis, and Phaseolus vulgaris. The rhizobial strain 11B was used successfully as a biofertilizer in agriculturally important legumes, forest trees, and agroindustrial plants.     

施硅对水稻白叶枯病抗性及叶片抗氧化酶活性的影响

【目的】水稻白叶枯病是一种细菌性枯萎病害,是限制水稻生产的重要生物因素之一。通过田间接种白叶枯病菌,研究施硅对水稻叶片中丙二醛含量及抗氧化系统酶活性的影响及其抗白叶枯病的机理,为安全有效的防治病害提供理论依据。【方法】以唐粳2号水稻品种为材料,2013年在河北省秦皇岛市进行田间试验,试验在两个施氮水平 [N 180 kg/hm2(正常供氮,N180), 450 kg/hm2(高量供氮,N450)]下设3个硅处理[不施硅(-Si),施硅酸钠(Si1, 以SiO2计,70 kg/hm2), 施硅钙肥(Si2, 以SiO2计,70 kg/hm2)],在水稻孕穗期采用剪叶法接种白叶枯病菌,研究硅对接种后30 d水稻病情指数和第1 d、 3 d、 5 d、 7 d和10 d水稻叶片中丙二醛(MDA)含量、 超氧化物歧化酶(SOD)活性、 过氧化氢酶(CAT)活性、 抗坏血酸过氧化物酶(APX)活性的影响。【结果】接种白叶枯病菌后,正常供氮水平,施硅处理的病情指数比不施硅处理平均降低17.8%(P0.05); 高量供氮水平,施硅钙肥的病情指数比不施硅降低15.1%(P0.05),而施硅酸钠的病情指数差异不显著。接种白叶枯病菌后,施硅处理的水稻叶片MDA均低于不施硅处理,且在正常供氮水平第7 d和高量供氮水平第3 d、 第7 d差异达显著水平。接种白叶枯病菌后,正常供氮水平第1 d、 第7 d和高量供氮水平第1 d、 第5 d,施硅处理的水稻叶片中SOD活性均显著高于不施硅处理,且第1 d施硅钙肥的叶片SOD显著高于施硅酸钠处理; 接种白叶枯病菌后,施硅处理的水稻叶片中CAT活性均高于不施硅处理,但未达显著水平; 高量供氮水平第1 d、 第7 d和第10 d施硅处理的水稻叶片中APX活性均显著高于不施硅处理。【结论】施硅能提高感病水稻叶片中SOD、 CAT和APX的活性,降低水稻叶片中MDA含量,有效清除植物体内活性氧(ROS),从而增强了水稻抗白叶枯病的能力; 在高量供氮水平下,硅钙肥抵御白叶枯病效果好于硅酸钠。     

水稻白叶枯病再流行原因分析与防控对策研究

为分析水稻白叶枯病历史演变与流行原因,研讨综合防治技术,对中国1960—2014年和浙江省温州市1971—2015年水稻白叶枯病的发生情况进行调查统计和分析。结果发现20世纪70—90年代初期为该病重发流行阶段,90年代中后期为间歇流行阶段,进入21世纪后病害发生减轻,但近年又趋上升态势。2014、2015年浙江省晚稻白叶枯病发病点多面广,浙江省东南沿海和沿江沿溪部分稻区发病较为严重,造成水稻产量较大损失。对该病在浙江省的发生动态及其影响流行的相关因子解析表明感病品种的种植与抗性退化、台风暴雨频繁、病原菌的累积和栽培管理措施不当是导致近年病害再度上升流行的主因;提出了新的耕作栽培条件下病害的防控对策,集成了一套以监测预警预报、选用抗（耐）病良种为基础,杜绝病菌来源为前提,秧田防治为重点,健身栽培和合理肥水、本田适期施药预防保护的综合防治技术,在生产上示范推广应用,有效控制该病的发生和流行。     

微生态制剂对养殖后期虾池水质及细菌群落的影响

水产养殖可持续发展的要求促进了微生态制剂的研究,为了解微生态制剂MP\|1和MP\|2的应用效果及其作用机理,选择处于养殖后期虾池开展应用试验,每周从试验池中采样分析水体CODMn、氨氮、亚硝酸盐、硝酸盐和总氮的变化,并用PCR\|DGGE分析了虾池细菌群落结构的变化。结果显示：投加微生态制剂MP\|1和MP\|2后1周,虾池亚硝酸盐均低于001 mg·L-1,2周后虾池氨氮分别稳定在026～033 mg·L-1和035～048 mg·L-1,试验过程中水体硝酸盐和CODMn保持稳定,总氮的去除率达到341％以上;虾池的主要细菌类群和细菌群落结构发生显著变化。结果表明：微生态制剂MP\|1和MP\|2可显著改善养殖后期虾池水质,其净水作用可能是微生态制剂调控虾池土著微生物群落变化后的共同作用结果。     

Evaluating the protective efficacy of antigen combinations against Photobacterium damselae ssp. piscicida infections in cobia,Rachycentron canadum L.

Cobia, Rachycentron canadum L., is a very important aquatic fish that faces the risk of infection with the bacterial pathogen Photobacterium damselae ssp. piscicida, and there are few protective approaches available that use multiple antigens. In the present study, potent bivalent antigens from P. damselae ssp. piscicida showed more efficient protection than did single antigens used in isolation. In preparations of three antigens that included recombinant heat shock protein 60 (rHSP60), recombinant α‐enolase (rENOLASE) and recombinant glyceraldehyde‐3‐phosphate dehydrogenase (rGAPDH), we analysed the doses that elicited the best immune responses and found that this occurred at a total of 30 μg of antigen per fish. Subsequently, vaccination of fish with rHSP60, rENOLASE and rGAPDH achieved 46.9, 52 and 25% relative per cent survival (RPS), respectively. In addition, bivalent subunit vaccines – combination I (rHSP60 + rENOLASE), combination II (rENOLASE + rGAPDH) and combination III (rHSP60 + rGAPDH) – were administered and the RPS in these groups (65.6, 64.0 and 48.4%, respectively), was higher than that achieved with single‐antigen administration. Finally, in combination IV, the trivalent vaccine rHSP60 + rENOLASE + rGAPDH, the RPS was 1.6%. Taken together, our results suggest that combinations of two antigens may achieve a better efficiency than monovalent or trivalent antigens, and this may provide new insights into pathogen prevention strategies.     

Frost promotes the pathogenicity of Pseudomonas syringae pv. actinidiae in Actinidia chinensis and A. deliciosa plants

Frost occurs in all major areas of cultivation, presenting a threat for the production of kiwifruit crops worldwide. A series of experiments were performed on 1‐year‐old, potted plants or excised twigs of Actinidia chinensis and A. deliciosa to verify whether strict relationships exist between bacterial canker outbreaks from Pseudomonas syringae pv. actinidiae (Psa) attacks and the occurrence of autumn and winter frost events. The association between the occurrence of autumn frost and the sudden outbreak of bacterial canker in A. chinensis in central Italy has been confirmed. Both autumn and winter frosts promote Psa multiplication in the inoculated twigs of both species. The day after the frost, reddish exudates oozing from the inoculation sites were consistently observed in both species, and Psa was re‐isolated in some cases. During the thawing of both A. deliciosa and A. chinensis twigs, the 2‐cm upward and downward migration of Psa from the inoculation site was observed within 3 min, and the leaves were consistently colonized with the pathogen. A consistent brown discoloration, accompanied with a sour‐sap odour, was observed throughout the length of the excised twigs of both Actinidia species after Psa inoculation and winter frost. Psa inoculation induced a remarkably higher necrosis in excised twigs that were not frozen compared with P. s. pv. syringae inoculation. Antifreeze protection using irrigation sprinklers did not influence the short‐term period of Psa and P. s. pv. syringae multiplication in both A. deliciosa and A. chinensis twigs. Thus, the damage from frost, freeze thawing and the accumulation of Psa in Actinidia twigs promotes the migration of the pathogen within and between the orchards. Taken together, the results obtained in this study confirmed that A. deliciosa is more frost tolerant than A. chinensis, autumn frosts are more dangerous to these crops than winter frosts, and in the absence of Psa, young kiwifruit plants remain sensitive to frost.     

Identification of genes required for the fitness of Rhodococcus equi during the infection of mice via signature-tagged transposon mutagenesis

Rhodococcus equi is a Gram-positive facultative intracellular bacterium that causes pyogranulomatous pneumonia in foals and immunocompromised people. In the present study, signature-tagged transposon mutagenesis was applied for the negative selection of R. equi mutants that cannot survive in vivo. Twenty-five distinguishable plasmid-transposon (plasposon) vectors by polymerase chain reaction (PCR), each containing a unique oligonucleotide tag, were constructed and used to select the transposon mutants that have in vivo fitness defects using a mouse systemic infection model. Of the 4,560 transposon mutants, 102 mutants were isolated via a real-time PCR-based screening as the mutants were unable to survive in the mouse model. Finally, 50 single transposon insertion sites were determined via the self-cloning strategy. The insertion of the transposon was seen on the virulence plasmid in 15 of the 50 mutants, whereas the remaining 35 mutants had the insertion of transposon on the chromosome. The chromosomal mutants contained transposon insertions in genes involved in cellular metabolism, DNA repair and recombination, gene regulation, non-ribosomal peptide synthesis, and unknown functions. Additionally, seven of the chromosomal mutants showed a reduced ability to multiply in the macrophages in vitro. In this study, we have identified several biosynthetic pathways as fitness factors associated with the growth within macrophages and survival in mice.     

海南省芒果细菌性黑斑病原菌生理生化特性初步研究

为了明确海南省芒果细菌性黑斑病原菌的生理生化特性,为该病害的科学防治提供理论基础。从海南省4 县市分离获得12 株芒果细菌性黑斑病原菌(Xanthomonas campestris pv. mangiferaeindicae)菌株并对其进行13 项生理和7 项生化特性测定。结果表明,在所测试的13 项生理特征中有9 项[耐盐性（1%～3%）、pH（5.0～8.0）、吐温80、醋酸盐、蔗糖、α-半乳糖、乳糖、麦芽糖和甘露醇]反应结果完全相同,但对果糖、葡萄糖、鼠李糖和木糖等碳源的利用却有明显差异。在7 项生化试验项目中,除HNMG15 产H2S试验为阴性外,其余供试菌株的氧化酶、过氧化氢酶、淀粉酶、硝酸盐还原、淀粉水解、明胶水解及产硫化氢等生化试验结果则完全一致。利用NTSYS-pc 2.10 软件采用Jaccard’s 相似系数和UPGMA方法对12株供试菌株的生理生化特征进行聚类分析,发现不同地理来源的芒果细菌性黑斑病原菌菌株可以聚成一类,由此可以说明海南芒果细菌性黑斑病原菌在生理生化特性方面存在一定的差异。