An effective biocontrol bioformulation against Phytophthora blight of pepper using growth mixtures of combined chitinolytic bacteria under different field conditions

Phytophthora blight of pepper caused by Phytophthora capsici has devastating consequences when combined with other pathogens, including Rhizoctonia solani, Fusarium oxysporum, and Fusarium solani. In order to develop a field-effective biocontrol strategy against Phytophthora blight of pepper, three chitinolytic bacteria, Serratia plymuthica strain C-1, strongly antagonistic to P. capsici, Chromobacterium sp. strain C-61, strongly antagonistic to R. solani, and Lysobacter enzymogenes strain C-3, antagonistic to R. solani and Fusarium spp., were selected. In pot studies, application of cultures combining the three bacterial strains effectively suppressed Phytophthora blight more than application of any single bacterial strain. Bioformulations developed from growth of the strains in a simple medium containing chitin under large batch conditions resulted in effective control in field applications. Efficacy of the bioformulated product depended on both the dose and timing of application. Although the undiluted product suppressed Phytophthora blight under all field conditions, a 10-fold diluted product was effective in solar-sterilized greenhouses and in fields with crop rotation. These results suggest that the developed product could be a new effective system to control Phytophthora blight disease in pepper.     

Fusarium wilt on sweet basil: Cause and sources in southeastern Spain

This study concerned a new disease detected in 1997 in southeastern Spain — Fusarium wilt in basil (Ocimum basilicum L.), caused byFusarium oxysporum f.sp.basilici. Its importance was evaluated at two locations in the Almería area, where 14% of the plants presented symptoms of the disease after 4 months of cropping. The search for sources of the disease inoculum was centered on the health of the seeds and the polypropylene trays that were reused for plant production. Analysis of four lots of seeds from Germany and Italy showed that two of them harboredF. oxysporum f.sp.basilici. This finding was confirmed by the analysis of seeds collected from diseased plants. Furthermore, analysis of three reused trays revealed the presence of the pathogen on them and it was concluded that the trays acted as the source of dispersion of the mycosis. http://www.phytoparasitica.org posting July 14, 2004.     

The potential for Fusarium oxysporum f. sp. fragariae,cause of fusarium wilt of strawberry,to colonize organic matter in soil and persist through anaerobic soil disinfestation

Fusarium wilt of strawberry, caused by Fusarium oxysporum f. sp. fragariae, is a disease of primary concern for strawberry production in many countries. Crop rotation and anaerobic soil disinfestation (ASD) have gained recent interest for their potential to contribute to management of this disease. Both techniques involve incorporation of organic matter into soil, which may be utilized by strains of Fusarium that are competitive saprophytes. We show that F. oxysporum f. sp. fragariae can colonize strawberry, lettuce, raspberry, and broccoli leaf tissues, which are sources of organic matter generated during crop rotation. This pathogen increased in soil population density during ASD treatments that did not become anaerobic, possibly as a result of growth on the organic amendment. However, significant population decreases were observed after ASD treatment when at least 100,000 cumulative reduced mV hours occurred in a 14-day experiment. Post-ASD abundance of F. oxysporum f. sp. fragariae in soil was negatively correlated with cumulative reduced mV hours. The only treatment that consistently caused disinfestation was exposed to a maximum temperature of 22 °C, which indicates there is potential for developing effective ASD treatments in the cool climates where strawberries are grown. Awareness that F. oxysporum f. sp. fragariae can act as a competitive soil saprophyte should be further investigated for its potential to alter disease outcomes where organic amendments are applied.     

Fusarium wilt of basil in Greece: Foliar infection and cultivar evaluation for resistance

Fusarium wilt of basil (Ocimum basilicum), caused byFusarium oxysporum f.sp.basilici, is reported for the first time in Greece. Foliage inoculation of young plants resulted in a downward movement of the pathogen to the crown and roots and 20–30% plant mortality. Of 14 commercial basil cultivars evaluated for possible disease resistance using young plants, six out of eight large-leaved cultivars were found resistant, while all six small-leaved ones were susceptible. http://www.phytoparasitica.org posting Feb. 23, 2004.     

Survival of plant pathogens under structural solarization

Structural solarization of greenhouses is a nonchemical sanitation procedure. The method involves dry heating, since maximal temperatures may exceed 60°C and consequent relative humidity (r.h.) is low (ca 15%), under fluctuating temperature and r.h. regimes. Thirty-five structural solarization experiments were performed over 7 years, testing one bacterial and five fungal plant pathogens. Various aspects of pathogen thermal inactivation under this method were studied. Thermal inactivation of the various pathogens differed according to the organism and inoculum form. Sensitivity to heat was highest withClavibacter michiganensis subsp.michiganensis and lowest withFusarium oxysporum f.sp.radicislycopersici inoculum in dry infected tomato stems, with ED₈₀ values of 7 and 47 days, respectively; intermediate values were obtained forPythium sp.,F. oxysporum f.sp.melonis, F. oxysporum f.sp.basilici andSclerotium rolfsii. The maximal ambient temperatures were in the range of 28.2° to 33.1°C. Structural solarization for sanitation can be a useful component of integrated pest management in greenhouses. http://www.phytoparasitica.org posting Sept. 28, 2004.     

Identification,pathogenicity and community dynamics of fungi and oomycetes associated with pea root rot in northern France

The pea root rot complex is a major concern for green pea production worldwide. This study aimed at characterizing its composition and dynamics throughout a cropping season in northern France. To this end, fungi and oomycetes were isolated from green pea plant roots with symptoms sampled at the flowering stage in 22 fields in 2017, and at the pea emergence, elongation and flowering stages in two fields in 2018. Out of 646 isolates collected, 317 were identified using molecular markers. Fusarium oxysporum, F. solani and F. redolens were highly predominant. Pathogenicity tests separated the isolates into four aggressiveness groups. F. solani isolates were the most aggressive. Phylogenetic analysis of their TEF1 sequences showed that they mainly belonged to the F. pisi lineage, and that F. oxysporum isolates were genetically close to isolates from the UK that did not belong to the forma specialis pisi. In addition, several Clonostachys rhizophaga isolates are reported for the first time to cause pea root rot. The oomycetes were rarely found and were represented by a few Pythium spp. isolates. Lastly, this study shows that the fungal and oomycete communities associated with pea root rot change during the cropping season. The level of dissimilarity of the root-rot-associated communities decreased throughout the cropping season towards a more similar composition at the flowering stage, dominated by F. solani, F. oxysporum and F. redolens. The proportion of nonpathogenic to weakly pathogenic isolates decreased progressively during the growing season in favour of moderately to highly pathogenic isolates.     

Characterization of Fusarium population associated with wilt of jojoba in Israel

Fusarium wilt, caused by Fusarium oxysporum, is a major disease of jojoba, causing serious economic losses. This study was aimed at characterizing the Fusarium populations associated with jojoba in Israel. Fifty Fusarium isolates used in this study included 23 isolates from the 1990s (“past”) and 27 recently isolated (“recent”). All the isolates were characterized by arbitrarily primed (ap)-PCR and 16 representatives were additionally delineated using multilocus (tef1, rpb1, rpb2) phylogeny and evaluated for their pathogenic potential. Consequently, 88% of the isolates were identified and characterized to the F. oxysporum species complex. The remaining 12% grouped within the F. fujikuroi, F. solani, and F. redolens species complexes. Variations in the infection rate (16.7%–100%), disease symptoms (0.08–1.25, on a scale of 0–3), and fungal colonization index (0.67–2.17, on a scale of 0–4) were observed within the tested isolates, with no significant differences between the past and recent isolates. The representative isolates were assigned to 11 groups based on ap-PCR. Pathogenicity tests showed that isolates from Groups II, IV, and V were the most aggressive, whereas isolates from Groups III, VIII, and IX were the least aggressive. Among the tested isolates, F. oxysporum sensu lato was the most aggressive, followed by F. proliferatum, while F. nygamai was the least aggressive. This study demonstrates the complexity and genetic diversity of Fusarium wilt on jojoba in Israel, indicating possible multiple introductions of infected germplasm into the country.     

Allelism of theFcu-1 andFoc genes conferring resistance to fusarium wilt in cucumber

The inheritance of resistance toFusarium oxysporum f.sp.cucumerinum race 1 was determined in the cucumber cv. WIS-248 by analyzing segregation of F₁, F₂, and BC populations of crosses with the susceptible cv. Straight-8. Resistance was conferred by a single dominant gene. In an allelism test, it was proven that theFcu-1 gene, which confers resistance toF. oxysporum f.sp.cucumerinum races 1 and 2 in cucumber cv. SMR-18 and theFoc gene, which confers resistance toF. oxysporum f.sp.cucumerinum race 2 in cucumber cv. WIS-248, are indistinguishable.     

Inhibition of plant pathogens in vitro and in vivo with essential oil and organic extracts of Cestrum nocturnum L.

The efficacy of the essential oil and various organic extracts from flowers of Cestrum nocturnum L. was evaluated for controlling the growth of some important phytopathogenic fungi. The oil (1000 ppm) and the organic extracts (1500 μg/disc) revealed antifungal effects against Botrytis cinerea, Colletotrichum capsici, Fusarium oxysporum, Fusarium solani, Phytophthora capsici, Rhizoctonia solani and Sclerotinia sclerotiorum in the growth inhibition range of 59.2-80.6% and 46.6-78.9%, respectively, and their MIC values were ranged from 62.5 to 500 and 125 to 1000 μg/mL. The essential oil had a remarkable effect on spore germination of all the plant pathogens with concentration and time-dependent kinetic inhibition of P. capsici. Further, the oil displayed remarkable in vivo antifungal effect up to 82.4-100% disease suppression efficacy on greenhouse-grown pepper plants. The results obtained from this study may contribute to the development of new antifungal agents to protect the crops from fungal diseases.     

番茄茎叶提取物对8种植物病原菌的生物活性初步研究

采用6种溶剂对番茄茎叶进行平行提取,以葡萄白腐病菌、葡萄黑痘病菌、苹果腐烂病菌、苹果轮纹病菌、苹果斑点落叶病菌、棉花枯萎病菌、瓜类枯萎病菌和番茄早疫病菌为供试菌,对番茄不同提取液进行抑菌活性测定。结果表明,各溶剂提取液对特定病菌都有极好的抑制作用,对葡萄白腐病菌和苹果腐烂病菌的抑制率均为100%,对其它病菌也都有很好的活性。番茄茎叶的水提取液对供试8种病原菌的综合抑制效果最好,对葡萄白腐病菌、葡萄黑痘病菌和苹果腐烂病菌的抑制作用高达100%,对瓜类枯萎病菌抑菌效果最差也为67.74%。,因此,水应作为番茄茎叶农用抑菌活性物质提取的首选溶剂,石油醚可作为有机溶剂提取番茄茎叶农用抑菌活性物质的溶剂。     

晋南地区黄瓜、黑籽南瓜死秧的病原菌鉴定及品种抗性研究

经分离、培养对不同菌种培养性状的观察,确定了侵染黄瓜、黑籽南瓜造成死秧的镰刀菌主要为尖镰孢菌黄瓜专化型、尖镰孢菌西瓜专化型、串珠镰刀菌和腐皮镰孢菌4种。经致病性测定,4种镰刀菌均能侵染黄瓜,引起发病造成死秧,可分为强致病类型和中强致病类型。经抗病性鉴定,黑籽南瓜种子只有南瓜4号为耐病品种;黄瓜种子也只有津优31号为耐病品种。     

枸杞炭疽病生防菌株筛选、生物功能测定及防治效果

为研发对枸杞炭疽病有良好防治效果的生防制剂,利用枸杞炭疽病菌Colletotrichum acutatum对实验室已分离保存的芽胞杆菌菌株进行室内筛选,并对拮抗效果较好的菌株进行形态学和分子生物学鉴定、病原菌孢子萌发抑制试验、生物学功能测定、稳定性测定、抑菌谱测定以及室内离体防治效果试验和田间防治效果试验。结果表明,菌株F3A对枸杞炭疽病菌有较好的拮抗作用,对枸杞炭疽病菌菌丝的抑制率为62.13%;结合形态学特征、16S rDNA以及gyrA基因序列分析,将菌株F3A鉴定为贝莱斯芽胞杆菌Bacillus velezensis;菌株F3A具有溶解有机磷和无机磷的能力,在含色氨酸和不含色氨酸的金氏培养液中产吲哚乙酸量分别为5.76 mg/L和5.74 mg/L;菌株F3A产蛋白酶和葡聚糖酶的活性较高;菌株F3A连续培养20代后,对枸杞炭疽病菌的抑制率为61.21%,该菌对棉花枯萎病菌Fusarium oxysporum f.sp.vesinfectum、番茄早疫病菌Alternaria solani、黄瓜枯萎病菌F.oxysporum f.sp.cucumerinum、番茄灰霉病菌Botrytis cinerea、番茄叶霉病菌Cladosporium fulvum、茄子菌核病菌Sclerotinia sclerotiorum和黄芪根腐病菌F.solani的抑制率分别为40.71%、53.58%、32.00%、53.00%、79.27%、71.13%和66.08%;菌株F3A发酵液的保护作用明显优于治疗作用,菌株F3A发酵液1倍稀释液室内预防效果为90.32%;喷施菌株F3A发酵液1倍稀释液3 d和7 d后的田间防治效果分别为78.26%和63.19%。表明菌株F3A有作为开发枸杞炭疽病生物农药的潜力。     

OS-诱抗剂诱导植物抗病性的作用探讨

用PDA培养基平板法测定了0.4%OS-诱抗剂水剂对水稻纹枯病菌、小麦纹枯病菌、油菜菌核病菌、辣椒立枯病菌、瓜类绵腐病菌、黄瓜枯萎病菌的生物活性,其EC₅₀值分别为34.56、59.33、33.17、85.92、91.91、122.87μg/mL,OS-诱抗剂对水稻纹枯病、油菜菌核病较好。高效液相色谱分析表明,经OS-诱抗剂处理后的植物提取液中酚类物质的种类和含量相对于对照有明显的变化,说明OS-诱抗剂对植物的防病作用可能是促使植物体内产生了酚类抗病物质。     

PCR-based differentiation of Fusarium oxysporum ff. sp. lycopersici and radicis-lycopersici and races of F. oxysporum f. sp. lycopersici

The pathogenic type (form and race) of Fusarium oxysporum, which generates wilt symptoms on tomato, was rapidly identified with a polymerase chain reaction (PCR)-based technique. We compared the partial nucleotide sequences of endo polygalacturonase (pg1) and exo polygalacturonase (pgx4) genes from isolates of F. oxysporum ff. sp. lycopersici (FOL) and radicis-lycopersici (FORL) from Japan and designed specific primer sets (uni, sp13, sp23, and sprl) based on the nucleotide differences that appeared among the pathogenic types. PCR with the uni primer set amplified a 670∼672-bp fragment from all isolates of FOL and FORL. With the sp13 primer set, an amplicon of 445 bp was obtained only from isolates of FOL race 1 and 3. With the sp23 primer set, a 518-bp fragment was obtained from isolates of FOL race 2 and 3. The sprl primer set yielded a 947-bp fragment from isolates of FORL, but not from FOL. A combination of amplifications with these primer sets effectively differentiated the pathogenic types of F. oxysporum in tomato.     

Fusarium Redolens f.sp asparagi, Causal Agent of Asparagus Root Rot, Crown Rot and Spear Rot

Two Fusarium species, F. oxysporum f.sp. asparagi and F. proliferatum, are known to be involved in the root and crown rot complex of asparagus. We have investigated reports on the involvement of F. redolens, a third species, which until recently was considered conspecific with F. oxysporum because of morphological similarities. RFLP analysis of the rDNA internal transcribed spacer region and AFLP fingerprinting identified eight strains from asparagus unambiguously as F. redolens. Four of these were tested and found to be pathogenic to asparagus either in this study (two strains) or in a previous one in which they were classified as F. oxysporum (three strains). Disease symptoms and disease development were the same as with F. oxysporum f.sp. asparagi and F. proliferatum. Present data and literature reports identify F. redolens as a host-specific pathogen involved in root, crown and spear rot of asparagus. The pathogen is formally classified as F. redolens Wollenw. f.sp. asparagi Baayen.     

Molecular identification and pathogenicity of Fusarium and Alternaria species associated with root rot disease of wolfberry in Gansu and Ningxia provinces,China

Wolfberry (Lycium barbarum) ranks in the top 10 best-selling medicinal plants in China and it has been used for centuries as a medicine and a food supplement. It is suggested to have benefits on human health due to the rich content of polysaccharides, carotenoids, flavonoids, and alkaloids contained in its fruits, leaves, and root bark. Recently, severe root rot diseases have been causing plant losses in major growing areas. Here, we report fungi causing root rot disease in Chinese wolfberry plants. The analysis of nucleotide sequences of the internal transcribed spacer (ITS) region revealed a total of 92 isolates isolated from both soil and plant material samples. Fusarium spp. were the most abundant (58%), followed by Penicillium spp. (9%), and Alternaria spp. (5%). Fusarium spp. included F. oxysporum (36%), F. solani (30%), F. chlamydosporum (9%), F. nematophilum (9%), and F. tricinctum (8%). Sequences from the translation elongation factor 1-α gene (TEF-1α) were used to confirm the identity of Fusarium spp. and showed the predominance of F. oxysporum and F. solani. To confirm the pathogenicity of isolates, four isolates belonging to Fusarium spp. and one isolate belonging to Alternaria spp., isolated from wolfberry root tissues with root rot symptoms, were tested in outdoor and laboratory conditions. Results revealed that the five tested isolates were pathogenic with varying degrees of aggressiveness and ability to induce symptoms of root rot in wolfberry seedlings. The five isolates were recovered from inoculated seedlings, completing Koch's postulates. This is the first report on causative agents of root rot in Chinese wolfberry.     

Simple diagnosis using ethanol immersion of strawberry plants with latent infection by Colletotrichum acutatum, Dendrophoma obscurans, and Fusarium oxysporum f. sp. fragariae

Simple diagnosis by ethanol immersion (SDEI) to detect Glomerella cingulata was used to detect three other fungi that also cause latent infection of strawberry plants. Signs on strawberry leaves with asymptomatic latent infection by Colletotrichum acutatum became visible using SDEI. Salmon-pink conidial masses were produced in the acervuli on the treated leaves 5 days after incubation at 28°C. In the case of Dendrophoma obscurans, pycnidia with amber conidial masses formed 5 days after incubation at 28°C. The pycnidia were observed mainly on the ribs, and conidial masses exuded from the ostiole. These macroscopic conidial masses were similar to those of G. cingulata and C. acutatum. When water was dripped onto a lesion caused by D. obscurans, the pycnidia exuded white filamentous conidial masses, making the distinction of D. obscurans from G. cingulata or C. acutatum. On petioles with latent infection by Fusarium oxysporum f. sp. fragariae, white aerial hyphae grew out from the vascular tissues on the cut surface 3 days after incubation at 28°C and were easily observed by eye or with a loupe. Thus, SDEI was also useful for diagnosing latent infection of strawberry plants by C. acutatum, D. obscurans, and F. oxysporum f. sp. fragariae.     

Genetic variability among isolates of Fusarium oxysporum from sugar beet

Fusarium yellows, caused by the soil‐borne fungus Fusarium oxysporum f. sp. betae (Fob), can lead to significant yield losses in sugar beet. This fungus is variable in pathogenicity, morphology, host range and symptom production, and is not a well characterized pathogen on sugar beet. From 1998 to 2003, 86 isolates of F. oxysporum and 20 other Fusarium species from sugar beet, along with four F. oxysporum isolates from dry bean and five from spinach, were obtained from diseased plants and characterized for pathogenicity to sugar beet. A group of sugar beet Fusarium isolates from different geographic areas (including nonpathogenic and pathogenic F. oxysporum, F. solani, F. proliferatum and F. avenaceum), F. oxysporum from dry bean and spinach, and Fusarium DNA from Europe were chosen for phylogenetic analysis. Sequence data from β‐ tubulin, EF1α and ITS DNA were used to examine whether Fusarium diversity is related to geographic origin and pathogenicity. Parsimony and Bayesian MCMC analyses of individual and combined datasets revealed no clades based on geographic origin and a single clade consisting exclusively of pathogens. The presence of FOB and nonpathogenic isolates in clades predominately made up of Fusarium species from sugar beet and other hosts indicates that F. oxysporum f. sp. betae is not monophyletic.     

Lack of biocontrol capacity in a non-pathogenic mutant of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">melonis</Emphasis>

The aim of this study was to assess the biocontrol capacity of rev157, a non-pathogenic mutant of a pathogenic strain of Fusarium oxysporum f. sp. melonis (Fom24). Inoculated in association with the virulent parental strain, the mutant rev157 did not protect the host plant (muskmelon) against infection by Fom24. Applied on flax, a non-host plant, the mutant rev157 was not able to protect it against its specific pathogen F. oxysporum f. sp. lini. On the contrary the parental strain Fom24 did protect flax as well as a soil-borne biocontrol strain (Fo47). Since the mutant rev157 was affected neither in its growth in vitro nor in its capacity to penetrate into the roots, it can be speculated that the mutation has affected traits responsible for interactions within the plant. In F. oxysporum the pair of strains Fom24/rev157 is a good candidate to identify genes involved in the biocontrol capacity of F. oxysporum and to test the hypothesis of a link between capacity to induce the disease and capacity to induce resistance in the plant.     

<Emphasis Type="Italic">In vivo</Emphasis> evaluation of essential oils and biocontrol agents combined with heat treatments on basil cv Genovese Gigante seeds against <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">basilici</Emphasis>

Seed treatments with essential oils (from savory and thyme) and biocontrol agents (Pseudomonas spp. and Fusarium oxysporum) have been evaluated in vivo after dry hot air treatments against Fusarium oxysporum f. sp. basilici on basil seeds. The savory and thyme essential oils showed a significant pathogen control activity because of their innate antifungal activity and because of the seed application method, but the dry hot pre-treatment did not show any obvious effect on the performance of the essential oil treatments. The dry heat treatment improved the Pseudomonas seed dressing effect against F.oxysporum f. sp. basilici, and showed important reductions in plant infection and the disease index on the treated seed plants, without any negative effect on seed germination. However, the pathogen control provided by the heat treatments combined with the application of the biocontrol agents never reached the same performance as the chemical treatments considered as the reference. Thus, short dry heat treatments on basil seeds have been shown to be a valid but complementary seed disinfection method against Fusarium wilt.