核桃黑斑病生防链霉菌YNF36的鉴定及其抑菌活性测定

【背景】由野油菜黄单胞菌(Xanthomonas campestris)和成团泛菌(Pantoea agglomerans)侵染引起的核桃黑斑病是一种严重的细菌性病害,给核桃产业带来了极大损失。【目的】从根际土壤中筛选出对核桃黑斑病病原菌野油菜黄单胞菌和成团泛菌均具有拮抗效果的放线菌菌株,可作为创制生防菌剂的出发菌株。【方法】采用稀释涂布法、平板对峙法和改良牛津杯法筛选拮抗菌株,通过形态学特征、生理生化特性和16S rRNA基因序列分析进行鉴定,测定无菌发酵液抗菌谱,离体叶片试验验证其对核桃黑斑病的防治效果。【结果】筛选到一株对2种病原菌均有较强拮抗作用的放线菌菌株YNF36。经形态学特征观察、生理生化特性试验及16S rRNA基因序列分析,将菌株YNF36鉴定为沙场链霉菌(Streptomyces arenae)。该菌株在SYP培养基上产量最高,抑菌活性最强,其无菌发酵液对金黄色葡萄球菌、大肠杆菌、黑曲霉、白色念珠菌、枯草芽孢杆菌、铜绿假单胞菌、蜡样芽孢杆菌这7种指示菌,以及链格孢菌、黑腐皮壳菌、胶孢炭疽菌、灰葡萄孢菌、黄褐孢霉菌、辣椒刺盘孢菌、腐皮镰孢菌这7种植物病原菌均有抑制作用,抗菌谱广。发酵液原液对离体叶片上的由野油菜黄单胞菌和成团泛菌造成的核桃黑斑病防效分别为75.69%和62.39%。【结论】沙场链霉菌YNF36补充了一种防治核桃黑斑病的生防材料,具有良好的开发价值和应用前景。     

Antagonistic and plant growth promoting activities of endophytic and soil actinomycetes

The objective of this study was the isolation and screening of actinomycete isolates for antagonistic potential and plant growth promoting activities. A total of 321 isolates were recovered from different plants, their rhizospheric soils and non-rhizospheric soils of Punjab and Himachal Pradesh regions. Out of these, 62 were endophytic, 156 were rhizospheric and 103 were non-rhizospheric isolates. In primary screening (dual culture assay), 83 isolates antagonised one or more test phytopathogenic fungi. From these active isolates, 20 were found to be antagonistic in well diffusion assay (secondary screening) and most of them demonstrated broad spectrum inhibitory activity against five to six test fungi. Studies on plant growth promoting activities revealed that 12 showed abilities to produce indole acetic acid, 10 produced siderophores and 12 showed ammonia production. Phosphate solubilisation was observed in five isolates and four fixed atmospheric N₂. In addition, production of hydrolytic enzymes such as chitinase, amylase, cellulase and protease was demonstrated by five, twenty, eleven and eleven isolates, respectively. The results of this study indicate that these isolates may be used as biocontrol and plant growth promoting agents. Morphological and chemotaxonomic studies revealed that all the active isolates belonged to the genus Streptomyces     

Greek indigenous streptomycetes as biocontrol agents against the soil‐borne fungal plant pathogen Rhizoctonia solani

Aims

To examine the biocontrol potential of multiactive Greek indigenous Streptomyces isolates carrying antifungal activity against Rhizoctonia solani that causes damping‐off symptoms on beans.

Methods and Results

A total of 605 Streptomyces isolates originated from 12 diverse Greek habitats were screened for antifungal activity against R. solani DSM843. Almost one‐third of the isolates proved to be antagonistic against the fungus. From the above isolates, six were selected due to their higher antifungal activity, identified by analysing their 16S rRNA gene sequence and studied further. The obtained data showed the following: firstly, the isolates ACTA1383 and ACTA1557 exhibited the highest antagonistic activity, and therefore, they were selected for in vivo experiments using bean seeds as target; secondly, in solid and liquid culture experiments under optimum antagonistic conditions, the medium extracts from the isolates OL80, ACTA1523, ACTA1551 and ACTA1522 suppressed the growth of the fungal mycelium, while extracts from ACTA 1383 and ACTA1557 did not show any activity.

Conclusions

These results corresponded important indications for the utility of two Greek indigenous Streptomyces isolates (ACTA1557 and ACTA1383) for the protection of the bean crops from R. solani damping‐off symptoms, while four of them (isolates OL80, ACTA1523, ACTA1551 and ACTA1522) seem to be promising producers of antifungal metabolites.

Significance and Impact of the Study

This is the first study on the biocontrol of R. solani using multiactive Streptomyces isolates originated from ecophysiologically special Greek habitats. Our study provides basic information to further explore managing strategies to control this critical disease.     

Isolation of Endophytic Actinomycetes From Roots and Leaves of Banana (Musa Acuminata) Plants and Their Activities Against Fusarium oxysporumf. sp. cubense

Two hundred and forty-two actinomycete strains were isolated from the interior of leaves and roots of healthy and wilting banana plants. Most of them were streptomycetes, Streptomyces griseorubiginosus-like strains were the most frequently isolated strains. Community analysis demonstrated increased actinomycete diversity in wilting leaves compared to that in healthy leaves, similar actinomycete communities were found in wilting and healthy roots. Screening of the isolates for antagonistic activity against Fusarium oxysporumf. sp. cubenserevealed that the proportion of antagonistic streptomycetes in healthy roots was higher than that in wilting roots (P < 0.01), but no difference was found between antagonistic strains isolated from healthy and wilting leaves. The potential biological control of Panama disease of banana by endophytic streptomycetes, especially Streptomyces griseorubiginosus-like strains was discussed.     

Influence of phylloplane colonizing biocontrol agents on the black spot of rose caused by Diplocarpon rosae

Abstract

An attempt was made to study the biocontrol efficacy of antagonistic microorganisms in phylloplane of rose cv. Edward to manage the black spot (Diplocarpon rosae) disease. Eight antagonistic microorganisms were tested in vivo against the black spot pathogen. Among these, Trichoderma viride and Pseudomonas fluorescens pf1 reduces the mycelial growth significantly. These two biocontrol agents were evaluated for their ability to induce defense-related enzymes and chemicals in plants. Increased activity of phenylalanine ammonia lyase (PAL), peroxidase (PO), polyphenoloxidase (PPO) and total phenolics were recorded in all the biocontrol agents treated leaves. P. fluorescens Pf1 recorded early and increased synthesis of the entire defense-related enzymes and total phenol within 6 days. The application of biocontrol agents induced the defense-related enzymes involved in phenyl propanoid pathway in addition to direct antagonism, which collectively contribute for enhanced resistance against invasion of Diplocarpon rosae in rose.     

一株抗茶炭疽菌和魔芋镰刀菌的淀粉酶产色链霉菌的分离、鉴定及生防潜能

【背景】放线菌是一类重要的生防菌,具有强大的代谢活性,能产生抗生素、酶、酶抑制剂和激素等天然产物抑制病原物生长。【目的】从茶树根际分离得到放线菌,研究候选放线菌对茶炭疽菌和魔芋镰刀菌的抑菌活性及其生防潜能。【方法】分别以茶炭疽病致病菌Colletotrichum camelliae和魔芋茎腐病致病菌Fusarium solani为指示菌,采用土壤稀释涂布法、平板对峙法和菌丝生长速率法,从茶树根际土壤中分离、筛选拮抗放线菌,并根据菌株的形态特征、生理生化特性和系统发育分析结果对其进行分类鉴定,并开展候选放线菌的产促生相关物质和分泌细胞壁水解酶能力的定性检测试验。【结果】共分离得到14株拮抗放线菌,菌株A-dyzsc04-2的拮抗效果最强,被鉴定为淀粉酶产色链霉菌（Streptomyces diastatochromogenes）。该菌株的活菌体对C. camelliae和F. solani的抑制率分别为66.71%±1.23%和71.59%±2.46%,其无菌发酵滤液对2种指示菌的抑菌率均大于90%;此外,菌株A-dyzsc04-2还具有产嗜铁素和葡聚糖酶以及溶解无机磷的能力。【结论】菌株A-dyzsc04-2是一株优良的生防菌,具有较高的开发利用价值,研究结果为菌株A-dyzsc04-2防治茶炭疽病和魔芋茎腐病提供了理论支撑。     

Epiphytic bacteria antagonistic to Curvularia leaf spot of yam

Curvularia eragrostidis yam leaf spot is a serious concern among the northeast Brazilian yam growing areas. In order to study its biocontrol, bacterial isolates from the yam phylloplane were tested against the pathogen. They were evaluated with respect to the following parameters: (1) inhibition of C. eragrostidis mycelial growth by using paired culture and cellophane membrane methods, (2) inhibition of conidium germination by using a paired suspension test, (3) reduction of disease severity and, (4) persistence of antagonistic action, on plants under greenhouse conditions. From a total of 162 bacterial isolates, 39 showed antagonism to the pathogen in paired culture. The bacteria produced extracellular, nonvolatile, and diffusible metabolites in the membrane cellophane test. Seventeen isolates resulted in more than 75% inhibition of C. eragrostidis mycelial growth. Among them, IF-26 showed the greatest antagonism. The isolates IF-82, IF-88, and IF-109 inhibited pathogen conidial germination, with average inhibition levels of 99.2, 98.2 and 96.2%, respectively. Under greenhouse conditions the antagonists were applied at three different time intervals relative to C. eragrostidis inoculation: 3 days before, at the same time, and 3 days after. IF-82 and IF-88 applied at the same time as pathogen inoculation both reduced disease severity 75%. IF-82 showed the best persistence of antagonistic action, with an average of 96.3%. IF-82, identified as Bacillus subtilis, was the best biocontrol agent for the yam leaf spot disease in this study.Correspondence to: Sami J. Michereff     

Two imide substances from a soil-isolated Streptomyces atratus strain provide effective biocontrol activity against grapevine downy mildew

Grapevine downy mildew (Plasmopara viticola) is a devastating disease of grapevine. In this study, 151 actinomycete isolates were obtained and tested for antagonistic activity against P. viticola. The assay suggested that 28 isolates displayed antagonistic effects to varying degrees. The greatest reduction in disease severity was observed with isolate PY-1, which reduced disease severity by 92.13% in the detached leaf assay, and by 83% in a field assay. It was identified as Streptomyces atratus using the 16S rDNA sequence analysis. To elucidate the antagonistic mechanism of PY-1 against P. viticola, scanning electron microscopy showed that major damage to the pathogens sporangia and sporangiophores was observed after treatment for PY-1. Furthermore, PY-1 showed antagonistic activity against other pathogens, including: Botrytis cinerea, Colletotrichum orbiculare, Fusarium oxysporum, Phytophthora capsici and Phytophthora infestans. Two imide compounds were purified from the fermentation liquid using silica gel chromatography and high-performance liquid chromatography and identified as 5-acetoxycycloheximide and cycloheximide using nuclear magnetic resonance. Both compounds showed significant antagonistic activity against P. viticola, determining a reduction in disease severity by 65% and 84%, respectively. In conclusion, 5-acetoxycycloheximide and cycloheximide were identified for the first time in a new S. atratus strain able to effectively control grapevine downy mildew.     

Application of Soil‐borne Actinomycetes for Biological Control against Fusarium Wilt of Chickpea (Cicer arietinum) caused by Fusarium solani fsp pisi

Black root rot, caused by Fusarium solani f.sp. pisi, is a devastating soil‐borne disease in chickpea in Iran with no effective control measures. With the aim of finding applicable biocontrol agents to alleviate the malady, isolates of Actinomycetes isolated from soil and their antagonistic effect against F. solani f.sp. pisi were evaluated both in vitro and in vivo. More than 100 Actinomycetes isolates were screened for their antifungal activities against the pathogen. The most active isolates were evaluated in greenhouse for their biocontrol performance. Based on the results of dual cultures in screening evaluations, the size of inhibition zone of fungal growth, and the most effective antagonist isolates (S3, S12 and S40) were selected for further studies. Identity of active isolates was determined, in this regard, 16S rDNA of isolates were amplified using universal bacterial primers FD1 and RP2. The PCR products were purified and sequenced. Sequence analysis of 16S rDNA was then performed using NCBI BLAST method. Comparison of the near full length 16S rRNA sequence of isolates to GenBank sequences demonstrated that isolates S3 and S12 were most similar to Streptomyces antibioticus, while isolate S40 was most similar to Streptomyces peruviensis. Biocontrol studies of these isolates in control of the disease in greenhouse significantly decreased the disease severity. Actinomycetes isolate S12 demonstrated the greatest effect in reducing disease than the other two. Results of this research are at preliminary stage for developing biocontrol agents. These data can be utilized as a platform for future studies with the aim of commercializing these biocontrol products and hoping to step towards sustainable agriculture.     

Biocontrol bacteria selected by a direct plant protection strategy against avocado white root rot show antagonism as a prevalent trait

Aim: This study was undertaken to study bacterial strains obtained directly for their efficient direct control of the avocado white root rot, thus avoiding prescreening by any other possible mechanism of biocontrol which could bias the selection. Methods and Results: A collection of 330 bacterial isolates was obtained from the roots and soil of healthy avocado trees. One hundred and forty‐three representative bacterial isolates were tested in an avocado/Rosellinia test system, resulting in 22 presumptive protective strains, all of them identified mainly as Pseudomonas and Bacillus species. These 22 candidate strains were screened in a more accurate biocontrol trial, confirming protection of some strains (4 out of the 22). Analyses of the potential bacterial traits involved in the biocontrol activity suggest that different traits could act jointly in the final biocontrol response, but any of these traits were neither sufficient nor generalized for all the active bacteria. All the protective strains selected were antagonistic against some fungal root pathogens. Conclusions: Diverse bacteria with biocontrol activity could be obtained by a direct plant protection strategy of selection. All the biocontrol strains finally selected in this work were antagonistic, showing that antagonism is a prevalent trait in the biocontrol bacteria selected by a direct plant protection strategy. Significance and Impact of the Study: This is the first report on the isolation of biocontrol bacterial strains using direct plant protection strategy in the system avocado/Rosellinia. Characterization of selected biocontrol bacterial strains obtained by a direct plant protection strategy showed that antagonism is a prevalent trait in the selected strains in this experimental system. This suggests that antagonism could be used as useful strategy to select biocontrol strains.     

甘蔗赤腐病内生拮抗菌YC89的筛选及鉴定

【目的】筛选防治甘蔗赤腐病(sugarcane red rot)的生防菌株。【方法】实验以前期分离获得的甘蔗内生细菌为目标菌,以甘蔗赤腐病的病原真菌镰孢炭疽菌(Colletotrichum falcatum Went.)为指示菌,采用平板对峙法筛选对该病菌有较强抑制作用的菌株,然后通过琼脂扩散法测定菌株代谢产物对抑菌活性的影响,并对具有较好拮抗效果的高效菌株进行抑菌广谱性分析并对其进行鉴定。最后通过形态学、生理生化特征以及16SrDNA和gyrA序列分析对高效菌株YC89进行鉴定。【结果】经初筛筛选到抑菌带均大于1.60 cm的5株拮抗细菌,其中X22、W2、YC89抑菌带均高达1.87 cm。对初筛得到的5株内生菌进行复筛,结果所示菌株YC89、H1、X22、W2、YT93对镰孢炭疽菌的抑菌率都在75%以上,其中菌株YC89对该病菌的抑菌效果最好,其抑菌率为78%。菌株YC89的发酵液、上清液、过滤液及粗蛋白提取液对镰孢炭疽菌的生长有较强的抑制作用,且菌株YC89对玉米大斑病、甘蔗梢腐病、草莓灰霉病等7种病原菌也有较好的抑制效果。通过菌株鉴定结果,初步将YC89菌株鉴定为贝莱斯芽孢杆菌(Bacillus velezensis)。【结论】菌株YC89对镰孢炭疽菌具有较好的抑制效果,表明其对甘蔗赤腐病的生物防治具有较大的应用潜力。     

Bio-formulation of fluorescent Pseudomonas spp. induces systemic resistance against red rot disease and enhances commercial sugar yield in sugarcane

Abstract

Isolates of Pseudomonas spp. collected from the rhizosphere of sugarcane and cane stalks were screened for their antagonistic activity against Colletotrichum falcatum causing red rot disease in sugarcane. Talc formulations of the selected Pseudomonas spp. isolates improved the sugarcane vegetative sett germination and sugarcane growth under field conditions. Optimal talc formulations were assessed for their effect on induction of systemic resistance against the pathogen in the canes under artificial inoculation. All the four isolates CHAO, EP1, KKM1 and VPT4 were effective in inducing systemic resistance against C. falcatum in two seasons. In other studies, the bacterial formulations were assessed to induce resistance in sugarcane in a sick plot situation. In pathogen-infested soil the isolates KKM1 and CHAO suppressed the red rot disease development in susceptible sugarcane cultivar. Pseudomonas strains also protected sugarcane in a disease-endemic location. Pseudomonas spp treatment substantially improved the cane juice quality parameters affected by the pathogen infection. Standardization of talc formulations and application methods in the field offers potential for large-scale application of biocontrol formulations for the management of red rot disease in sugarcane growing regions.     

番茄内生菌的分离鉴定及菌株FQ-G3抗病促生特性

由灰葡萄孢(Botrytis cinerea)引起的灰霉病是番茄生产中最重要的病害之一,当前使用的杀菌剂因药物残留、病原菌抗药性及食品安全等原因逐渐受到限制。因此,利用拮抗微生物的生物防治逐渐成为灰霉病防控的有效策略。【目的】从番茄植株体内筛选具有抗病促生特性内生菌株并对其生防潜力进行评估,为开发番茄灰霉病生物防治新策略提供理论依据。【方法】采用组织分离法在番茄植株不同部位分离出内生细菌、真菌,结合16SrRNA和ITS序列分析,对候选菌株进行初步鉴定;通过菌株对峙培养、果实离体接种筛选对灰葡萄孢具有拮抗活性的内生菌;进一步测定菌株分泌生长素、嗜铁素的能力及其对拟南芥和番茄幼苗生长的促生特性。【结果】从番茄植株不同部位共分离出72株内生细菌和31株内生真菌,通过平板对峙法筛选出1株对多种病原菌具有较好抑菌活性的内生细菌FQ-G3,分子鉴定为Bacillus velezensis。FQ-G3对灰葡萄孢抑菌率达80.93%,并显著抑制灰葡萄孢在番茄果实上的扩展。该菌株能够分泌生长素、蛋白酶和嗜铁素,且对拟南芥、番茄幼苗具有明显的促生效果。【结论】本研究表明分离自番茄植株的内生菌FQ-G3具...     

Evaluation of Streptomyces spp. and Bacillus spp. for biocontrol of Fusarium wilt in chickpea (Cicer arietinum L.)

A study was carried out to test direct and indirect antagonistic effect against Fusarium wilt, caused by Fusarium oxysporum f. sp. ciceri (FOC), and plant growth-promoting (PGP) traits of bacteria isolated from rhizosphere soils of chickpea (Cicer arietinum L.). A total of 40 bacterial isolates were tested for their antagonistic activity against FOC and of which 10 were found to have strong antagonistic potential. These were found to be Streptomyces spp. (five isolates) and Bacillus spp. (five isolates) in the morphological and biochemical characterisation and 16S rDNA analysis. Under both greenhouse and wilt sick field conditions, the selected Streptomyces and Bacillus isolates reduced disease incidence and delayed expression of symptoms of disease, over the non-inoculated control. The PGP ability of the isolates such as nodule number, nodule weight, shoot weight, root weight, grain yield and stover yield were also demonstrated under greenhouse and field conditions over the non-inoculated control. Among the ten isolates, Streptomyces sp. AC-19 and Bacillus sp. BS-20 were found to have more potential for biocontrol of FOC and PGP in chickpea. This investigation indicates that the selected Streptomyces and Bacillus isolates have the potential to control Fusarium wilt disease and to promote plant growth in chickpea.     

Design,Synthesis and Antibacterial Activity of Novel Pyrimidine-Containing 4H-Chromen-4-One Derivatives**

A series of pyrimidine-containing 4H-chromen-4-one derivatives were designed and synthesized by combining bioactive substructures. Preliminary biological activity results showed that most of the compounds displayed significant inhibitory activities in vitro against Xanthomonas axonopodis pv. Citri (X. axonopodis), Xanthomonas oryzae pv. oryzae (X. oryzae) and Ralstonia solanacearum (R. solanacearum). In particular, compound 2-[(3-{[5,7-dimethoxy-4-oxo-2-(3,4,5-trimethoxyphenyl)-4H-1-benzopyran-3-yl]oxy}propyl)sulfanyl]-4-(4-methylphenyl)-6-oxo-1,6-dihydropyrimidine-5-carbonitrile ( 4c ) demonstrated a good inhibitory effect against X. axonopodis and X. oryzae, with the half-maximal effective concentration (EC₅₀) values of 15.5 and 14.9 μg/mL, respectively, and compound 2-[(3-{[5,7-Dimethoxy-4-oxo-2-(3,4,5-trimethoxyphenyl)-4H-1-benzopyran-3-yl]oxy}propyl)sulfanyl]-4-(3-fluorophenyl)-6-oxo-1,6-dihydropyrimidine-5-carbonitrile ( 4h ) showed the best antibacterial activity against R. solanacearum with an EC₅₀ value of 14.7 μg/mL. These results were better than commercial reagents bismerthiazol (BT, 51.7, 70.1 and 52.7 μg/mL, respectively) and thiodiazole copper (TC, 77.9, 95.8 and 72.1 μg/mL, respectively). In vivo antibacterial activity results indicated that compound 4c displayed better curative (42.4 %) and protective (49.2 %) activities for rice bacterial leaf blight than BT (35.2, 39.1 %) and TC (30.8, 27.3 %). The mechanism of compound 4c against X. oryzae was analyzed through scanning electron microscopy (SEM). These results indicated that pyrimidine-containing 4H-chromen-4-one derivatives have important value in the research of new agrochemicals.     

Biological control of Botrytis cinerea on tomato using naturally occurring fungal antagonists

Abstract

In order to evaluate the potential of naturally occurring filamentous fungi having potential as biocontrol agents effective against grey mould and post-harvest fruit rot caused by Botrytis cinerea on tomato, fungal saprophytes were isolated. They were obtained from leaves, fruits and flowers belonging to different species of cultivated and spontaneous Solanaceous plants collected at the horticultural area of La Plata, Argentina. Of 300 isolates screened for inhibition of B. cinerea using the dual culture technique on agar plate, 12 strains inhibited strongly mycelial growth of the pathogen. Among the antagonists one isolate of Epicoccun nigrum (126), four of Trichoderma harzianum (110, 118, 248 and 252) and four isolates of Fusarium spp. decreased the spore germination of B. cinerea between 30 and 70%. These isolates were probed on tomato fruits to evaluate their biocontrol activity against post-harvest grey mould. In growth chamber tests, E. nigrum (27), F. equiseti (22, 105) and T. harzianum (118, 252) reduced the diameter of fruit lesions by 50 – 90% and were selected for further biocontrol assays of tomato plants in the greenhouse. Although there were not significant differences between the treatments and the control, F. equiseti (105), E. nigrum (27) and T. harzianum (118) reduced by 20, 22 and 22 respectively the disease on whole plants. The targeted application of isolates of E. nigrum, T. harzianum and F. equiseti provides a promising alternative to the use of fungicide spray to control B. cinerea on tomatoes.     

Antifungal activities of actinomycete strains associated with high-altitude sagebrush rhizosphere

The antifungal-producing potential of actinomycete populations from the rhizosphere of low-altitude sagebrush, Artemisia tridentata, has been examined. In a continued investigation of new sources of antifungal-producing microorganisms, this study examined the antifungal-producing potential of actinomycetes from the rhizosphere of high-altitude A. tridentata. With high-altitude sagebrush, rhizosphere soil actinomycete numbers were one to four orders of magnitude higher than those found in nonrhizosphere bulk soils and different from those found with the low-altitude plants. A total of 122 actinomycete isolates was screened against nine fungal species and six bacterial species for the production of antimicrobial compounds. Four rhizosphere isolates, Streptomyces amakusaensis, S. coeruleorubidus, S. hawaiiensis and S. scabies, showed broad-spectrum antifungal activity against three or more fungal species in plate assays. In liquid antagonism assays, mycelium production by Aspergillus niger was reduced by up to 50% by two of the actinomycete isolates. These results demonstrate the potential of rhizosphere microbiology in the search for new antimicrobials.     

Genetic and phenotypic characterization of Xanthomonas axonopodis pv. maculifoliigardeniae causing bacterial leaf spot of Ixora in Taiwan

Ixora spp. are Rubiaceae plants commonly planted as hedges or potted flower. Recently, incidents of bacterial leaf spot of Ixora were observed in central parts of Taiwan. Previous research on the disease has been scarce and focused mainly on its diagnosis. Therefore, many characteristics of the causal agent remain unclear. The present study aims to improve our understanding of this lesser-characterized pathogen and provide information useful for its identification and management. Bacterial strains Ixo1, Ixo2 and Ixo3 were isolated from infected Ixora x westii. All three isolates were able to grow and induce leaf spot symptoms on Ixora. They also exhibited morphological and physiological characteristics typical of Xanthomonads. Biolog analysis indicated that Ixo1 to Ixo3 have metabolic fingerprints similar to X. axonopodis pv. poinsettiicola. Multilocus sequence analysis and inoculation assays identified Ixo1 to Ixo3 as X. axonopodis pv. maculifoliigardeniae, albeit their gene sequences were very similar to other species/pathovars belonging to the X. euvesicatoria species complex; members of this species complex have different plant hosts, yet share similar housekeeping gene sequences. A semi-specific PCR assay evaluated in this work was able to differentiate Ixo1 to Ixo3 from bacteria not belonging to the X. euvesicatoria species complex, suggesting that the assay may be used in diagnosing bacterial leaf spot of ixoras. Finally, the sensitivity of the isolated pathogen to multiple commercial pesticides was tested, and the results showed that the bacterium is sensitive to streptomycin + tetracycline (10％ SP), thiophanate methyl + streptomycin (68.8％ WP) and oxolinic acid (20% WP), but more tolerant against copper-based chemicals. Overall, the findings from this work may facilitate the identification and management of X. axonopodis pv. maculifoliigardeniae.     

Plant growth regulators and signal molecules enhance resistance against bacterial blight disease of pomegranate

Bacterial blight caused by Xanthomonas axonopodis pv. punicae is a devastating disease causing huge economic losses in major pomegranate growing areas. Thus, this study has been undertaken to assess the effect of plant growth regulators (PGRs) and signal molecules on disease incidence of bacterial blight in addition to its impact on yield and productivity under field condition in disease endemic area. Considering the environmental concern for reduction in synthetics and antibiotics, PGRs offers cost‐effective and an eco‐friendly schedule for management of this disease. Foliar application of ethylene at 200 ppm resulted in significant reduction in disease incidence in leaves (from 33.01% to 4.87%), fruits (from 25.77% to 2.05%) and twigs (from 1.76% and 0.97% to 0%), as compared to untreated plants. Treatment with salicylic acid and paclobutrazol also resulted in reduction of disease incidence. Further, higher accumulation of total phenols and anthocyanin content in rind as well as in arils was observed in ethylene treatment with enhanced fruit yield in pomegranate plants. The enhanced disease tolerance by recording less disease incidence with increased accumulation of defence compounds like phenols and anthocyanins by ethylene, salicylic acid and paclobutrazol treatments resulted in improving fruit yield and quality of pomegranate fruits. Thus, judicious application of PGRs and signal molecules contributed for induction of resistance while improving developmental and reproductive parameters of pomegranate and thus could be a viable option for their synergistic use in pomegranate farming.     

Attachment microbes antagonistic against Botrytis cinerea—biological control and scanning electron microscope studies in vivo

Bacteria and yeasts, selected by an attachment assay for their ability to adhere to Botrytis cinerea hyphae or conidia, were evaluated for biocontrol potential against B. cinerea on excised tomato stems. Eight of the 12 bacteria and seven of the eight yeast isolates conferred 90% to 100% biocontrol activity when antagonist populations were applied at three to 80 times the pathogen inoculum density. Biocontrol was maintained at similar levels when biocontrol agent (BCA) application was delayed up to 48 h after pathogen challenge. Scanning electron microscopy showed extensive colonisation of B. cinerea mycelium or conidia by many of these isolates and also evidence of pathogen degradation. The biocontrol efficacy and potential for the assay are discussed with respect to cell-to-cell adhesion as a vehicle to deliver antagonistic mechanisms to highly specific pathogen sites.