Lime and ammonium carbonate fumigation coupled with bio-organic fertilizer application steered banana rhizosphere to assemble a unique microbiome against Panama disease

Microbiome plays a key role in determining soil suppressiveness against invading pathogens. Our previous study revealed that microbial community of bulk soil could be manipulated by lime and ammonium bicarbonate fumigation followed by biofertilizer application. However, the assembly of microbial community suppressive to banana Panama disease in the rhizosphere is still unclear. In this study, we used high-throughput sequencing and quantitative PCR to explore the assembly of rhizosphere microbiome associated with banana Panama disease suppression in a two-seasonal pot experiment. We found biofertilizer applied to lime and ammonium bicarbonate fumigated soil significantly (P < 0.05) reduced the abundance of rhizosphere Fusarium oxysporum compared to biofertilizer applied to non-fumigated soil. Principal coordinate analysis revealed that biofertilizer applied to lime and ammonium bicarbonate fumigated soil re-shaped the rhizosphere bacterial community composition by increasing the phylogenetic relatedness, and stimulating indigenous microbes, for example, Gemmatimonas, Sphingomonas, Pseudomonas, Lysobacter and Bacillus. Co-occurrence analysis revealed that potential species involved in disease suppression were more interrelated in disease-suppressive soils. Taken together, lime and ammonium bicarbonate fumigation followed by biofertilizer application could induce banana rhizosphere to assemble beneficial microbes dominated consortia to suppress banana Panama disease.     

施肥处理对不同抗性品种香蕉枯萎病的防控效果

【目的】随着香蕉枯萎病菌4号生理小种热带型(简称Foc TR4))在云南的入侵、传播和蔓延,对云南的香蕉产业产生严重的威胁。通过实时荧光定量PCR分析蕉园定植香蕉后7个月内的土壤中枯萎病病原菌TR4含量动态变化,明确不同香蕉品种的大田抗性表现以及不同肥料的防控效果,为枯萎病的防控提供技术参考。【方法】选用巴西蕉、桂蕉1号、南天黄和自主选育的云蕉1号为供试品种开展田间试验,设置虾肽有机肥+虾肽特护+虾肽果叶康(简称:虾肽有机肥处理)、常规有机肥+微生物制剂(简称:微生物处理)和常规有机肥(简称:对照)3个处理,调查4个品种在4个时间段的枯萎病发病率和3种肥料的防治效果。【结果】在月平均枯萎病病原菌TR4含量均超过2000拷贝的土壤条件下,4个品种的发病率在3个施肥处理中均表现出差异性,南天黄、云蕉1号的发病率与其他2个主栽感病品种的发病率差异达显著水平;3种施肥处理间的发病率达显著差异,发病率从高到低表现为对照虾肽有机肥处理微生物处理。【结论】施用微生物制剂对降低枯萎病发病率起一定的作用。南天黄的抗病性较强,云蕉1号也表现出较强的抗性,但还有待进一步改良和提高抗性。     

米修链霉菌TF78对香蕉枯萎病的田间防效及根际土壤微生物的影响

【背景】香蕉枯萎病是香蕉生产上的毁灭性病害,生物防治是遏制该病害发生的有效手段。在前期的研究中,从健康香蕉根际土壤中分离获得一株对香蕉枯萎病具有良好盆栽防治效果的生防菌——米修链霉菌(Streptomyces misionensis) TF78,但其对香蕉枯萎病的田间生防潜力和对土壤微生物环境的影响尚不清楚。【目的】评价米修链霉菌TF78对香蕉枯萎病的田间防治效果,明确其对香蕉根际土壤微生物群落的影响。【方法】选取两块发病香蕉园,测定该生防菌株对香蕉枯萎病的防治效果,并利用扩增子测序技术分析施用菌剂组和空白对照组共12份香蕉根际土壤的微生物多样性和丰度。【结果】米修链霉菌TF78对两块香蕉园的田间防效分别达55.30%和45.32%。该生防菌株处理组的物种稀释曲线坡度大于空白对照组,并显著富集了优势种群梳霉门(Kickxellomycota),消减了绿弯菌门(Chloroflexi)、酸杆菌门(Acidobacteria)和苔藓杆菌(Bryobacter)的丰度,对土壤中优势种群变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)、放线菌门(Actinobacteria)、芽单胞菌门(Gemmatimonadetes)及木霉属(Trichoderma)、鞘氨醇单胞菌属(Sphingomonas)、寡养单胞菌属(Stenotrophomonas)和芽孢杆菌属(Bacillus)的相对丰度影响不显著。【结论】米修链霉菌TF78塑造了不利于香蕉枯萎病菌Fusarium oxysporum f.sp.cubense存活的土壤环境,有效降低了田间香蕉枯萎病的发生,同时对土壤中大部分具有重要生态功能和抑菌功能的优势微生物种群影响不显著。该研究结果为米修链霉菌TF78的进一步开发应用奠定了基础。     

Effects of rhizosphere soil,vesicular-arbuscular mycorrhizal fungi and phosphate onPlantago major L. ssp.pleiosperma Pilger

Biotic factors in the rhizosphere and their effect on the growth ofPlantago major L. ssp.pleiosperma Pilger (Great plantain) were studied. In a pot experiment the effect on shoot growth of the addition of 2.5% rhizosphere soil at four levels of phosphate was highly dependent on the availability of phosphate: a promoting effect at low phosphate levels was observed while a reducing effect occurred at higher phosphate levels. As the roots were infected with vesicular-arbuscular mycorrhizal (VAM) fungi in the treatment with rhizosphere soil, two other experiments were set up to separate effects of the indigenous VAM fungi from effects of the total rhizosphere population. The uptake of phosphate and shoot growth was not decreased at higher phosphate availability when VAM inoculum was added alone or in combination with rhizosphere soil. The growth reducing effect of the rhizosphere soil could therefore not be ascribed only to mycorrhizal infection. The results suggest that biotic factors in the rhizosphere soil affect the phosphate uptake ofPlantago major ssp.pleiosperma. This may, under conditions of phosphate limitation, lead to an increase of phosphate stress and, subsequently, a growth reduction. Futhermore, it is concluded that VAM fungi, as part of the rhizosphere population, may compensate this phosphate stress by enhancing the phosphate uptake.Grassland Species Research Group Publication No. 148.     

Studies on the rhizosphere mycoflora of broad bean and cotton

Summary The fungal flora of the rhizosphere of three varieties of broad bean and cotton was studied by the dilution-plate technique. The numbers of fungi were higher in the rhizosphere than in the non-rhizosphere soil. Plant type and age, and soil type have a significant influence of the nature and numbers of fungal flora associated with plant roots.Cladosporium was relatively more abundant in the rhizosphere of broad bean varieties, whilePenicillium was found to constitute a high percentage of fungi found in the rhizosphere of cotton varieties. Plant variety has no influence on the nature of such fungal flora.     

Comparative study on the host susceptibility of different banana cultivars to plant parasitic nematodes in Malappuram district of Kerala,India

Six cultivars of banana grown widely in Malappuram district of Kerala were evaluated for the diversity of plant parasitic nematodes in their respective rhizosphere. The genotypes evaluated include Nendran, Poovan (Mysore), Njali poovan, Mysore poovan, Chenkadali and Ponnan. The study aims on a comparative analysis of rhizosphere of different banana varieties for the occurrence of plant parasitic nematodes, diversity, frequency of occurrence and to determine the most vulnerable type of banana variety. Cobb’s decanting and sieving method was used to isolate nematodes from soil. Thirty three samples randomly collected from different blocks of Malappuram district are subjected to analysis and a ten species of nematodes were reported. Among the six varieties analysed both Nendran and Mysore Poovan variety seems to be more susceptible to phytoparasitic nematodes. With respect to eight blocks of Malappuram district, Helicotylenchus sp. shows highest prominence value followed by Radopholus similis, reniform nematode and Meloidogyne incognita.     

A new bioorganic fertilizer can effectively control banana wilt by strong colonization with Bacillus subtilis N11

Fusarium wilt is one of the most serious diseases caused by a soil-borne pathogen affecting banana production. The goal of this study was to evaluate the capability of a novel bio-organic fertilizer (BIO2) that integrated the biocontrol agent Bacillus subtilis N11, and mature composts to control Fusarium wilt of banana in pot experiments. The results showed that the application of the BIO2 significantly decreased the incidence rate of Fusarium wilt compared to the control. To determine the antagonistic mechanism of the strain, we also studied the colonization of the natural biocontrol agent on banana roots using a GFP marker. The studies were performed in a hydroponic culture system, a sand system and a natural soil system. The results indicated that the bacteria colonized predominantly by forming biofilms along the elongation and differentiation zones of the roots. The fact that similar observations were obtained in all three systems suggests that colonization by N11 can be studied in a defined system. The population of B. subtilis N11 in the rhizosphere and on banana roots was also monitored. We speculate that the colonization pattern of B.subtilis N11 can be linked to the mechanism of protection of plants from fungal infection.     

Fusarium species on groundnut kernels and in groundnut soil

Summary A total of 17 species and varieties of Fusarium have been isolated from groundnut kernels, rhizosphere and geocarposhere and from the soil of groundnut fields in Israel.F.solani predominated in the Fusarium flora of soil, rhizosphere and fresh kernels, and in particular on stored kernels. In the geocarposhere,F. solani, F. oxysporum andF. equiseti occurred in fairly equal amounts. Fusaria made up 16.2 to 32.6 per cent of the total mycoflora of kernels, soil, rhizosphere and geocarposphere.In tests with 14 isolates from groundnuts, production of toxins inducing visible reactions on rabbit skin was strongest when cultures were kept at 24 to 30°C. Only one isolate failed to produce a toxic reaction.The mortality of seedlings induced in inoculation tests with 23 groundnut isolates ofF. oxysporum, F. solani andF. equiseti was severe (mostly averaging 20 to 40%) in tomato and eggplant, moderate (mostly 10 to 20%) in bean, cucumber, watermelon and onion, weaker in cotton and pepper, and least in maize and wheat.     

Nitrate reduction in the leaves and numbers of nitrifiers in the rhizosphere ofPlantago lanceolata growing in two contrasting sites

Summary Numbers of autotrophic nitrifiers in the rhizosphere, and thein vivo nitrate reductase activity (NRA) in the leaves of individual plants ofPlantago lanceolata were determined in plants at two contrasting sites. In a dune grassland, high numbers of nitrifiers were present in the rhizosphere, and significant NRA was detected in the leaves. During dry periods nitrate utilization sometimes was depressed. In a wet hayfield, on peat soil, very low numbers of nitrifiers were found in the rhizosphere. Also the NRA was low. In the wet habitat, the NRA in the leaves of some fen species, containing aerenchyma in the roots, was higher than that ofP. lanceolata, not containing aerenchyma.Grassland Species Research Group. Publication No. 105.     

生姜根系不同生态位细菌群落多样性特征、组成及结构差异

生姜作为常见的调味品和传统中药材,是我国重要的经济作物之一。作为取食部分的生姜块茎与根系直接相连,其产量、品质与根相关细菌群落密切相关。然而,关于生姜根系微环境中细菌群落的特点仍鲜有报道,土壤环境能否衍生出宿主特异性内生菌群落尚不清楚。以生姜根系不同生态位细菌群落为研究对象,采用高通量测序技术,对非根际、根际及根内细菌进行16S rRNA基因测序。结果表明,不同生态位细菌群落多样性存在显著差异,其中非根际及根际细菌群落多样性(Shannon index, Observed species, Faith′s PD)显著高于内生菌群落。同时,各生态位共现网络稳定性和复杂度表现为非根际>根际>根内细菌群落。而在组成上,细菌群落在不同生态位差异显著(R²=0.57,P=0.001)。其中变形菌门(Proteobacteria)是根内的优势门,该门类下假单胞菌属(Pseudomonas)、短波单胞菌属(Brevundimonas)、寡养单胞菌属(Stenotrophomonas)及泛菌属(Pantoea)在根内显著富集。在根际细菌中,拟杆菌门(Bacteroid...     

Contribution to the knowledge of soil fungi in Libya

Summary Rhizosphere and soil counts of fungi aroundArtemisia herba alba grown in three localities in Tripoli have shown a rhizosphere effect. Counts from localities near the sea shore were higher than from areas a distant to the south. However, R/S ratios were almost the same in the three localities and on all the experimental agar media. Ten genera and 25 species commonly occurring in the rhizosphere or soil ofArtemisia could be identified. The order of occurrence for the chief genera of the rhizosphere ofArtemisia was the same in the three localities.     

Differential Colonization Patterns of Bananas (Musa spp.) by Physiological Race 1 and Race 4 Isolates of Fusarium oxysporum f.sp. cubense

Fusarium oxysporum f.sp. cubense (Foc) is the causative agent of Fusarium wilt of bananas (Musa spp.). To clarify the colonization patterns of Foc in bananas, two green fluorescent protein‐tagged isolates, NT320 (race 1) and B2‐gfp (race 4), were used to follow infection of the banana varieties Pisang Awak and Brazil. Penetration and colonization of both isolates in roots of these two banana varieties were observed within 6 days, but sporulation in xylem vessels was not observed until day 30 postinoculation. Interestingly, B2‐gfp penetrated into xylem vessels of Pisang Awak banana roots more quickly than NT320, implying that the race 4 isolate is more virulent than the race 1 isolate. This result was further confirmed by comparing the disease severity of plants inoculated with NT320 with that of plants inoculated with B2‐gfp. Quantitative real‐time PCR revealed that some pathogenicity‐associated genes, including Fga1, Fhk1, Fow2 and Ste12, were upregulated by B2‐gfp during exposure to Brazil bananas, while they were either downregulated by NT320 or not significantly changed. These data might partly explain why the race 4 isolate was more virulent than the race 1 isolate.     

广东省本地油茶和引种油茶根际土壤微生物群落特征

【背景】近年来,油茶低效林面积较大,根际土壤微生物影响林木抗性和生长,对林业可持续发展具有重要意义。【目的】了解广东省本地油茶和引种油茶根际土壤微生物群落特征。【方法】利用高通量测序分析油茶根际土壤微生物群落组成。【结果】油茶根际土壤细菌有26门77纲201目377科593属676种,真菌有14门50纲121目266科502属631种。油茶根际土壤中的优势细菌为酸杆菌门和变形菌门,优势真菌为子囊菌门和担子菌门。两种油茶根际土壤微生物组成差异显著,本地油茶根际土壤的细菌多样性显著高于引种油茶。在门水平上,脱硫杆菌门细菌和罗兹菌门、被孢霉门真菌的相对丰度在两种油茶间差异显著,Amorphotheca在本地油茶根际土壤中特异性富集。两种油茶根际土壤细菌碳代谢相对丰度差异显著,真菌以腐生营养型为主,其次为病理营养型和共生营养型。本地油茶根际土壤中显著富集土壤腐生菌,而共生营养型真菌(尤其是丛枝菌根真菌)相对丰度(6.43%)显著低于引种油茶中(21.83%)。此外,有机质和养分含量是影响油茶根际土壤微生物群落的关键因子。【结论】本地油茶和引种油茶根际土壤微生物群落组成和结构差异显著,Amorp...     

18份广东香蕉种质对枯萎病的抗性评价

【背景】香蕉枯萎病是世界性的香蕉毁灭性病害,尚无有效药剂防控,筛选抗病品种是目前理想的防治方法。【方法】采用组培苗伤根接种法,研究了18份香蕉种质对香蕉枯萎病菌4号生理小种的抗性水平,并根据病情指数进行抗性分级。【结果】在供试的18份香蕉种质中,2份(东莞大蕉、抗枯5号)高抗,2份(碧盛、大丰)抗病,3份(抗枯1号、粉杂、农科1号)中抗,7份(粤优抗1号、广东-741、泰国B9、大蕉、台湾8号、海贡蕉、威廉斯8818)感病,4份(巴西、广东2号、广粉1号、粉蕉)高感。【结论与意义】不同香蕉种质对香蕉枯萎病菌4号生理小种的抗病性存在较大差异,本研究初步筛选出7份抗枯萎病的香蕉种质,为香蕉枯萎病抗病育种提供了依据,为病区种植香蕉品种提供了有效参考。     

Survival ofPseudomonas syringae pv.lachrymans with cucumber roots

Summary Survival ofPseudomonas syringae pv.lachrymans with seedling cucumber roots, root washings, rhizosphere soil, and nonrhizosphere soil was determined 7–8 days after the soil surface was watered with a cell suspension of the bacterium. Plants were in pots in the green-house and soil was not sterilized. Survival was best with roots and root washings, next best in rhizosphere soil, and poor in nonrhizosphere soil.     

Induction of defence-related enzymes in susceptible variety of banana: role of Fusarium-derived elicitors

Elicitor prepared from the Fusarium oxysporum f. sp cubense (Foc) isolated from infected banana rhizosphere induced the accumulation of resistance-associated enzymes in leaves of susceptible and resistant variety of banana. Roots of Grand Naine (susceptible) and robusta (resistant) variety were inoculated with 1 g/l Foc elicitors. Distinct difference in peroxidase, polyphenol oxidase, β-1,3-glucanase, chitinase and phenolics was observed in control plants of resistant and susceptible varieties. Induced defence-related enzymes in susceptible variety were increased tothe level of untreated resistant variety. This depicted that Fusarium-derived elicitor effectively induced defence in susceptible variety to the apparent level of untreated resistant variety.     

Occurrence and correlations of nematodes,Fusarium oxysporum and edaphic factors on banana plantations

Problems caused by nematodes and Fusarium wilt (Panama disease) on banana plantations are responsible for yield losses and are limiting to its cultivation. In the state of Goias, there is little information about the nematode occurence on this crop, and its relation with the incidence of Fusarium oxysporum f.sp. cubense (Foc). This research had the purpose to identify the occurrence of plant‐parasitic nematodes on banana plantations and to verify its correlation with the Fusarium wilt and with the soil attributes (pH, texture, nutrients). Twelve banana orchards in the state of Goias were sampled in the municipalities of Anapolis, Caiaponia, Goiatuba, Itaguaru, Itumbiara (two areas), Jatai, Morrinhos, Ouro Verde, Palestina, Taquaral and Uruana. All sampled areas, except Morrinhos, revealed contamination with Foc, and all areas had different genera of nematodes either in the soil or in the roots samples. Meloidogyne sp., Helicotylenchus sp. and Rotylenchus sp. were the main genera of plant‐parasitic nematodes found in the samples, with Meloidogyne sp. and Rotylenchus sp. being the most dominant and abundant genera. The presence of Pratylenchus sp. increases the population levels of F. oxysporum. Helicotylenchus sp. is highly correlated with high concentrations of Mn. High population density of Meloidogyne sp. was found in irrigated areas with low concentrations of P, Ca, Mg and soil pH.     

Development of a mode of application of bioorganic fertilizer for improving the biocontrol efficacy to <Emphasis Type="Italic">Fusarium</Emphasis> wilt

More effective ways of applying biocontrol products should be developed based both on the characteristics of the biocontrol agents and the normal practices of the agricultural producer. A new system was developed to improve the biocontrol efficacy of Fusarium wilt for watermelon production, and this system was tested in pot and field experiments. Biocontrol was achieved by applying a novel bioorganic fertilizer product (BIO) to Fusarium-infested soil. The best biocontrol was obtained by application of a bioorganic fertilizer, BIO, into soil during the nursery phase of watermelon seedling followed by a second application to Fusarium-infested soil when watermelon seedlings were transplanted. In comparison with the controls, the incidence of the disease was reduced by 60–100% in the pot experiment and by 59–73% in the field experiment when the BIO was applied during the nursery stage. After application of BIO during the nursery stage, the number of colony-forming units of Fusarium oxysporum in rhizospheric soil was significantly (P < 0.05) inhibited compared to the controls. An in vitro experiment showed that the antagonist Paenibacillus polymyxa in the BIO could effectively colonize the rhizosphere of watermelon and proliferate along the extending plant roots. This inhibited growth of Fusarium oxysporum in the rhizosphere of watermelon and protected the watermelon roots from attack by the pathogens. The method used for biocontrol Fusarium wilt disease in watermelon should be a useful strategy to improve field efficacy of other biocontrol agents.     

Effect ofAgrobacterium radiobacter and its polysaccharide on emergence and damping-off of tomato plants

Agrobacterium radiobacter B6 and agrobacteran (an exopolysaccharide of the succinoglycan group) stimulated seed germination and tomato plant emergence. The germination was most stimulated by dipping the seeds in 0.1 % agrobacteran for 30 min whereas plant emergence in garden soil was best with 0.4 % agrobacteran at 10–20°C. Treatment of the plants withA.radiobacter cells and agrobacteran solution at 30–35°C. had a lower effect.A.radiobacter cells applied on seed surface protected the plants against damping off in garden soil artificially inoculated with the fungiRhizoctonia solani andPythium ultimum; in soils contaminated withFusarium solani 0.1 to 0.2% agrobacteran had a higher protective effect than the bacterization. The difference can be attributed to the varying density ofA.radiobacter population in plant rhizosphere in the presence of different plantpathogenic fungi, different interactions of microorganisms in the rhizosphere and different mode of penetration of the pathogens into plant roots.     

香蕉假茎生物炭对根际土壤细菌丰度和群落结构的影响

【目的】将香蕉假茎生物炭施加到土壤,探讨香蕉假茎生物炭对香蕉根际土壤微生物的影响。【方法】以香蕉假茎生物炭(BPB)0、1%、2%、3%的质量比与土壤均匀混合。盆栽培养3个月后分离香蕉根际土壤。采用16S rRNA高通量测序技术对根际土壤细菌群落结构和丰度进行表征。【结果】提高BPB施用量可增加土壤有机质、有效钾、有效磷含量,提高土壤pH值,但降低有效氮浓度。在1%BPB样品中获得2278个OTUs,其显示细菌群落中的最大多样性。施加3%的BPB处理土壤,拟杆菌门、疣微菌门和厚壁菌门的相对丰度显著增加;放线菌门、酸杆菌门、芽单胞菌门明显减少。主成分分析发现,1%BPB和2%BPB处理的样本之间土壤细菌群落相似。【结论】施加不同比例BPB改变了根际土壤中细菌丰度和群落结构,且高比例添加改变更加明显。