Inhibitive Activity of the Extracts of Plants in Artemisia against Fusarium oxysporum f. sp. vasinfectum and Fusarium momiliforme

[Objective]The aim was to provide theoretical basis for the control of cotton Fusarium wilt and cotton boll rot disease.[Method]The inhibitory activity of the extracts from three species of plants(Artemisia annual L.,Artemisia capillaris Thunb.,Artemisia argyi Levl.et Vant)against Fusarium oxysporum f.sp.vasinfectum and Fusarium moniliforme were studied under the condition of laboratory.[Result]The extracts of all the three plants in Artemisia showed strong antifungal activity against the tested pathogenic ...

棉花枯萎病菌(Fusarium oxysporum f.sp.Vasinfectum(ATK)Sndyder & Hansln的抑菌土初步研究

1987～1988年在河南新乡用相同接菌量种植感病品种,对棉花抗枯萎病品种86—1连作10年以上田块土样进行盆栽和田间小区试验。结果表明.枯萎病平均病指分别为29.4和4.4,与对照果园土比较抑菌效果分别为39.9％和85.8％。上述田块中86—1棉株根围5cm内土样,盆栽枯萎病平均病指19.5,抑菌效果60.5％。连作10年以上枯萎病圃土样,病指分别为27.2和6.8,抑菌效果为43.9％和76.2％。1990～1991年在辽宁省经作所试验结果,连作10年以上枯萎病圃田间抑菌效果56.2％,连作5年病圃抑菌效果17.6％。上述结果证明,枯萎病圃衰退的主要原因是棉花枯萎病菌抑菌土的产生和存在。土样内枯萎菌菌量测定结果表明,3个抑菌土土样内菌量比果园导菌土减少48.6％～59.2％。抑菌土经高压灭菌后抑菌效果完全消失,初步证明抑菌因子可能以生物因子为主。

从土壤中分离棉枯萎病菌(Fusarium oxysporum f.sp.vasinfetum (Atk) Synder & Hansen)选择性培养基研究

本文介绍一种用于分离检测土壤中棉枯萎病菌的选择性培养基—植选2号。其成分为:KH_2PO_41g,MgSO_4·7H_2O 0.5g,K_2S_2O_50.2g,KCl0.6g,NH_4NO_3 0.5g,蛋白胨5g,山梨糖10g,蔗糖5g,琼脂20g,蒸馏水1000ml,PCNB 620mg,Oxgall1g,硫酸链霉素300mg,盐酸金霉素75mg。根据棉枯萎病在此培养基上的形态特征,能较容易地识别和检测该病菌。

棉花枯萎病菌(Fusarium oxysporum f. sp. Vasinfectum)突变体的诱发与鉴定

通过对32个棉花枯萎病菌菌株进行的突变体诱发与鉴定实践及技术方法探索,进一步验证了可以采用KClO3培养基来诱发突变体,并利用各突变体对不同N源的利用能力差异而采用MM,MH,MA,MO2培养基对各突变体类型进行鉴定。实验结果显示绝大多数菌株(93.7%)在KClO3培养基上能诱发出突变体,但多数(85%)突变体类型不全,且不同菌株诱得突变体的难易不同,个别菌株很难诱发到突变体。各突变体的诱发率高低依次为nit1>nit3>nit8>nitM,与国内外相关报道结果基本一致。

应用巢式PCR检测黄瓜尖镰孢菌(Fusarium oxysporum f．sp．cucumbrum)

文章应用BIK2/BIK3和BIK1/BIK4两对引物,采用巢式PCR(Nested-PCR)技术对镰孢菌属26个菌株(代表了6个镰孢菌种)以及从黄瓜根际分离的20株真菌、6株细菌和7株放线菌共计59个菌株进行了扩增。结果显示,只有3个黄瓜尖镰孢菌的致病菌株能产生一条长度大约为800bp的片断。对接种黄瓜尖镰孢菌的黄瓜植株进行检测结果表明,巢式PCR能从接种5d后的黄瓜病样中特异性地检测出病原菌,而症状的出现需要10～13d时间。该项技术具有较高的灵敏度,适用于黄瓜枯萎病早期侵染检测研究。

DNA methylation patterns of banana leaves in response to Fusarium oxysporum f. sp. cubense tropical race 4

Fusarium wilt of banana, which is caused by Fusarium oxysporum f. sp. cubense tropical race 4 （Foc TR4）, is a serious soil-borne fungal disease. Now, the epigenetic molecular pathogenic basis is elusive. In this study, with methylation-sensitive amplification polymorphism （MSAP） technique, DNA methylation was compared between the leaves inoculated with Foc TR4 and the mock-inoculated leaves at different pathogenic stages. With 25 pairs of primers, 1 144 and 1 255 fragments were amplified from the infected and mock-inoculated leaves, respectively. DNA methylation was both changed and the average methylated CCGG sequences were 34.81 and 29.26% for the infected and the mock-inoculated leaves. And DNA hypermethylation and hypomethylation were induced by pathogen infection during all pathogenic stages. Further, 69 polymorphic fragments were sequenced and 29 of them showed sequence similarity to genes with known functions. And RT-PCR results of four genes indicated that their expression patterns were consistent with their methylation patterns. Our results suggest that DNA methylation plays important roles in pathogenic response to Foc TR4 for banana.

Biological Characteristics of Fusarium oxysporum f. sp.niveum Isolated from Watermelon Cultivated in Plastic Greenhouse and Indoor Toxicity Determination of Different Fungicides in Hainan Province of China

[ Objective] The paper was to study biological characteristics of Fusarium oxysporum f. sp. niveum in watermelon cultivated in plastic greenhouse in Hainan Province of China and to determine indoor toxicity of fungicides. [ Method] Effects of various factors on mycelial growth and spore germination were stud- ied, and the lethal temperature of spores and indoor toxicity of fungicides were also determined. [Result] The fungal mycelium had a better growth rate on water- melon juice medium than the others. The optimum temperature for mycelial growth and spore germination were 28 ℃ and 30 ℃, and the optimum pH values were 7 - 8 and 7 - 9, respectively. Light-dark alternation was conducive for mycelial growth. The lethal temperature of spore was 60 ℃ for 5 min. The fungal mycelium grew better while taking fructose and glucose as carbon sources and yeast extract and peptone as nitrogen sources. Toxicity determination showed that 50% pro- chloraz-manganese chloride complex WP had the greatest inhibition effect on mycelial growth, with the ECso value of 0.730 9 μg/mL; followed by 32.5% benzoic azoxystrobin SC, 30% benzoylate· propieanazol EC, 10% difenoconazole WG, 25% bromothalonil · carbendazim WP and 25% bromothalonil WP, with the ECso values from 1.884 7μg/mL to 8. 161 0μg/mL. [Conclusion] The study provided basic data for field control against F. oxysporum f. sp. niveum on watermelon cultivated in plastic greenhouse in Hainan Province.

Construction of PEG-mediated Genetic Transformation and Gene Knockout System in Fusarium oxysporum f. sp.cubense Tropic Race 4

Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense tropic race 4(Foc TR4), is a typical vascular and soil-borne disease which has significantly threatened the sustainable development of banana industry. In order to reveal the infection process and pathogenesis of Foc TR4, the young mycelia(66.7 mg/ml) of wild-type strain of Foc TR4(WT-Foc TR4) cultured for 18-20 h were lysed with enzyme mixture for protoplast formation, which consisted of 25 mg/ml driselase, 0.4 mg/ml chitinase, 15 mg/ml lysing enzyme and 1.2 mol/L potassium chloride. The resulted protoplasts of 2×10~7 cells/ml were used to test the efficiency of transformation mediated by polyethylene glycol, and up to 9 transformants per microgram of DNA were obtained. AmCyan, RFP and YFP genes were stably transferred into the WT-Foc TR4, separately, using the protoplast transformation system. The gene FoOCH1 encoding α-1, 6-mannosyltransferase in the WT-Foc TR4 was knocked out using the split-marker recombination technology. The genetic transformation and gene knockout system in this pathogen lays a foundation for the study of functional genomics and plant-pathogen interactions.

Biochemical and Physiological Changes of Three Watermelon Cultivars Infested with Fusarium oxysporum f.sp.niveum

The dynamic changes in membrane permeability, activities of disease-related enzymes, and contents of pathogenesis-relevant chemical compounds and photosynthetic pigments in root cells of three watermelon cultivars were studied after inoculation with Fusarium oxysporum f. sp. niveum at seedling stage. The results showed that the capacity of self-regulating and returning to normal status of cultivar Kelunsheng (resistant) was greater than that of a susceptible cultivar, Zaohua, in terms of malonaldehyde (MDA) content, relative conductivity, and activity of superoxide dismutase (SOD) and catalase (CAT). The resistant cultivar maintained a higher activity of dehydrogenase, higher content of vitamin C(Vc) , and relatively lower content of soluble sugar than the susceptible cultivar. The content of soluble protein was higher in the resistant cultivar than that in the susceptible one at day 1 after inoculation. The capacities of the resistant cultivar to inhibit chlorophyll deterioration and maintain a higher carotenoid content were significantly stronger than those of the susceptible cultivar.

Genetic analysis of the maximum germination distance of Striga under Fusarium oxysporum f.sp.strigae biocontrol in sorghum

Maximum germination distance（MGD） is an important component of Striga resistance in sorghum. The objective of this study was to determine gene action influencing MGD of Striga hermonthica and Striga asiatica among selected sorghum lines treated with a biocontrol agent, Fusarium oxysporum f. sp. strigae（FOS） for effective breeding with Striga resistance, and FOS compatibility. Twelve sorghum genotypes were selected based on their Striga resistance, FOS compatibility, and superior agronomic performance. Selected genotypes were crossed using a bi-parental mating design to generate six families for genetic analysis. Agar-gel assays were used to determine low haustorium initiation factor（LHF） using the 12 parental lines, their F_1 progenies, backcross derivatives, and F_2 segregants in two sets. One set had S. hermonthica seed and the other one had S. asiatica seed. Both were treated with and without FOS. Genotypes were evaluated using a split-plot design with three replications and MGD data were recorded followed by generation mean analysis. FOS reduced MGD by 1cm under both S. hermonthica and S. asiatica infestations. Additive, dominance, and epistatic gene actions were involved in the control of MGD of the two Striga species in the evaluated populations. On average, the relative contribution of additive, additive×additive and dominance×dominance genetic effects on the MGD of S. hermonthica and S. asiatica, with FOS, were 20, 33, and 36%; and 21, 32, and 35%, respectively. Breeding methods exploiting these genetic effects may provide enhanced response to selection for Striga resistance and FOS compatibility in integrated Striga management（ISM） programmes.

Physiological Races of Fusarium oxysporum f. sp. Ciceris in Iraq

Twenty isolates of Fusarium oxysporum f. sp. ciceris were isolated from wilted chickpea plants obtained from different districts of north part of Iraq to assess variability in pathogenicity of the populations. Each isolate was tested on 12 differential chickpea varieties. Isolates showed highly significant variation in wilt severity on the differential varieties. Based on the reaction types that induced on differential varieties, isolates were grouped into four groups, First group included isolates FocSl, FocQ7, FocQ 10, FocFI3, FocH 17 and FocHl8; the second group included isolates FocS2, FocS3, FocS4, FocQ5, FocQ8, FocQ9, FocF11, FocF12, FocFl4 and FocH19; the third group included isolates FocF15, FocHl6, FocH20; where the isolate FocQ6 was placed in the fourth group. Results showed that the percentage of genetic similarity was ranged 42% to 100% and was 42% between the first group and other groups and 72% between the three groups the rest and thus this indicate the presence of four races of the fungus which are O, 4, 5 and 1B/C, this represent the first record of these races in lraq.

Isolation and identification of Fusarium oxysporum f. sp. cubense in Fujian Province, China

Fusarium wilt, caused by Fusarium oxyporum f. sp. cubense(Foc), is the most serious disease affecting banana production.To clarify the distribution of the Foc races in Fujian Province of China, 79 soil samples were collected from four regions of Zhangzhou City, the primary banana production area in Fujian. We isolated and identified 12 Foc strains based on internal transcribed spacer(ITS) sequence analysis, PCR amplification by using Foc-specific primers and pathogenicity assays.Our analysis indicated that 11 isolates belong to Foc race 1, and 1 isolate belongs to the Foc tropical species race 4(TR4).Although TR4 has previously been reported to occur in primary banana-producing provinces, such as Hainan, Guangxi,and Guangdong of China, this is the first report of TR4 isolated from the soil in Fujian Province. Monitoring the presence of Foc, in particular, the TR4 strains in the soil, is the basic strategy to prevent and control Fusarium wilt.

Effects of Medium Nutrients on Fusarium oxysporum Schl. f.sp

[Objective] To study the effects of different culture conditions on the Fusarium oxysporurn SchL f. sp. [Method] Based on species identification of the pathogenic organism of Fusarium oxysporum Schl. f. sp, effects of different cultures and different nutrients on the mycelial growth and conidial production of Fusarium oxysporum SchL f. sp were studied. [Result] The mycelial growth and conidial pro- duction of Fusarium oxysporum SchL f. sp was different under different culture con- ditions. PDA medium was the most suitable medium for the mycelial growth and had the highest conidial production; and the mycelial grew the fastest on the medium with maltose as carbon source or peptone as nitrogen source, which also had the highest conidial production. [Conclusion] This study provided experimental basis for the study of Fusarium oxysporum SchL f. sp and also provided theoretical basis for the study and control of Fusarium oxysporum Schl. f. sp.

Isolation and Identification of Shoot-Tip Associated Endophytic Bacteria from Banana cv. Grand Naine and Testing for Antagonistic Activity against <i>Fusarium oxysporum</i>f. sp. <i>cubense</i>

Endophytic bacteria colonizing the shoot-tips of banana cv. Grand Naine were isolated and tested for the antagonistic activity against the Panama wilt pathogen Fusarium oxysporum f. sp. cubense (Foc). Pre-isolation, the suckers were given extensive disinfection treatments and the homogenate from the excised shoot-tip portion was plated on nutrient agar (NA) and trypticase soy agar (TSA). This yielded altogether 47 isolates: 26 on NA and 21 on TSA, respectively, from the 10 suckers collected during August to February. The number of bacterial isolates obtained per sucker varied from one to 15 based on colony characteristics registering up to 10 distinct species per shoot-tip based on 16S rRNA sequence analysis. The 47 isolates belonged to 19 genera and 25 species under the phylogenetic classes of Actinobacteria, α- and γ-Proteobacteria and Firmicutes. Actinobacteria constituted the predominant phylum (55% isolates) with the constituent genera of Arthrobacter, Brevibacterium, Corynebacterium, Curtobacterium, Kocuria, Kytococcus, Micrococcus, Naumanella, Rothia and Tessaracoccus spp. and an unidentified isolate belonging to the family Frankiaceae. Proteobacteria constituted the second major phylum (Brevundimonas, Enterobacter, Klebsiella, Pseudomonas, Serratia and Sphingomonas spp.) followed by Firmicutes (Bacillus and Staphylococcus spp.). Antagonistic activity of the endophytes against Foc was tested through agar plate assays (pit and spot applications on fungal lawn) employing potato dextrose agar and NA. Endophytic Pseudomonas aeruginosa (isolate GNS.13.2a) which was associated with a single sucker showed significant growth inhibition effect on Foc while Klebsiella variicola (GNS.13.3a) and Enterobacter cloacae (GNS13.4a) exhibited moderate inhibition. The study brings out considerable sucker to sucker variation in the associated cultivable endophytic bacteria in “Grand Naine” banana and identifies a few bacterial endophytes with biocontrol potential against the devastating Foc pathogen.

Characterization of Fusarium Oxysporum Isolates Obtained from Wax Gourd and Chieh-qua in China by Pathogenicity, RAMs and Sequence Analysis of the rDNA Internal Transcribed Spacers (ITS1 and ITS2)

Wax gourd (Benincasa hispida Thumb. Cogn) is called white gourd, winter melon, Chinese preserving melon, Chinese squash, and don kwa. It has been cultivated in China for over 2 300 years. It probably

Comparative study of <i>Fusarium oxysporum</i>f sp. <i>lycopersici</i>and <i>Meloidogyne incognita</i>race-2 on plant growth parameters of tomato

Many species of soil-inhabiting fungus Fusarium, cause severe yield loss in many crops. Experiments were conducted in net house condition with complete randomized block design to determine the individual effect of different in-oculum levels of root-knot nematode, Meloidogyne incognita, Race-2 and Fusarium oxysporum f sp. lycopersici on plant growth parameters viz., Plant length, fresh and dry weight and number of fruits of tomato var. P21. The experimental results showed that both the pathogens cause significant reduction in plant growth parameters. However, the fungus was not much effective on plant growth parameters in comparison to root-knot nematode. Greatest reduction in plant growth parameters was recorded in plants inoculated with 8000 J2/Kg soil of Meloidogyne in-cognita race 2. The threshold level of root-knot nematode was 1000 J2/kg soil while threshold level of Fusarium was @ 1 g/Kg soil. Inoculum level of Fusarium oxysporum f sp. lycopersici and Meloidogyne in-cognita race-2 was pathogenic and caused significant reduction at and above 1 g/kg soil and 1000 J2/Kg soil respectively.

A study on the pathogen species and physiological races of tomato Fusarium wilt in Shanxi,China

In order to clarify the main pathogens of tomato Fusarium wilt in Shanxi Province, China, morphological identification, elongation factor 1 alpha （EF-1α） sequence analysis, specific primer amplification and pathogenicity tests were applied to study the isolates which were recovered from diseased plants collected from 17 different districts of Shanxi Province. The results were as follows： 1） Through morphological and molecular identification, the following 7 species of Fusarium were identified： F. oxysporum, F. solani, F. verticillioides, F. subglutinans, F. chlamydosporum, F. sporotrichioides, and F. semitectum; 2） 56 isolates of F. oxysporum were identified using specific primer amplification, among which, 29, 5 and 6 isolates were respectively identified as F. oxysporum f. sp. lycopersici physiological race 1, race 2, and race 3; 3） pathogenicity test indicated the significant pathogenicity of F. oxysporum, F. solani, F. verticillioides, and F. subglutinans to tomato plant. Therefore, among these 4 species confirmed as pathogenic to tomato in Shanxi, the highest isolation rate （53.3％） corresponded to F. oxysporum. Three physiological species, race 1, race 2, and race 3 of F. oxysporum f. sp. lycopersici are detected in Shanxi, among which race 1 is the most widespread pathogen and is also considered as the predominant race.

Screening of Bacillus subtilis HAAS01 and Its Biocontrol Effect on Fusarium wilt in Sweet Potato

Fusarium wilt,a disease caused by Fusarium oxysporum f.sp batatas(Fob)is an important disease in sweet potato production.Using endophytic bacteria for biological control of sweet potato diseases is one of the important ways.A Bacillus subtilis with antagonistic effect on Fusarium wilt of sweet potato was isolated from soil by confrontation culture.According to the biological characteristics,16S rDNA sequence analysis,and physiological and biochemical analysis,the Bacillus subtilis HAAS01 was named.A pot experiment was conducted for the biological control experiment of strain HAAS01,and the endogenous hormone content,antioxidant enzyme activity,soluble protein content,and related gene expressions of sweet potato plants were detected.The results showed that the HAAS01 strain could promote the production of endogenous hormones and resist the infection of plant diseases together with defensive enzymes and upregulation of related gene expressions.In summary,Bacillus subtilis HAAS01 was effective in controlling Fusarium wilt of sweet potato and has potential for application and development.

Relationship between Tomato (Solanum lycopersicum) Nutrition and the Severity of the Vascular Fusariosis

The vascular fusariosis is one of the main obstacles in the tomato crop production. Currently, the management of the nutrient solution is presented as a control option for fusariosis, for such reason different nutrient solutions with nitrogen (N), phosphorus (P), potassium (K) and calcium (Ca) modifications;in four saladette tomato varieties Bony Best (BB), Manapal (M), Walter (W) and FLA were evaluated in this research, in order to analyze the damage caused by Fusarium oxysporum f.sp. lycopersici (FOL) severity. Tomato plants were inoculated with 3 FOL breeds, establishing a completely random design with a factorial arrangement with six repetitions. Disease’s severity was evaluated 30 days after the inoculation using levels of (0 - 4) of the methodology established by Marlatt and Correll (1988). Afterwards, a statistical analysis was done through the Kruskal Wallis test, in which it was observed that treatments 93 (Ca 207 mg∙L-1, R3, variety BB), 95 (207 mg∙L-1 Ca, R3, variety W), 81 (0Ca, R3, BB) showed less disease severity. In the foliar analysis, the best treatments were 22 (193 mg∙L-1 N, R3, M) for N, the greatest P content was treatment 94 (207 mg∙L-1 Ca, R3, M) for K treatment three and 93 (207 mg∙L-1 Ca, R3, variety BB) greatest Ca content.

The effects of Trichoderma on preventing cucumber fusarium wilt and regulating cucumber physiology

In our previous studies, we identified 3 Trichoderma strains with anti-Fusarium oxysporum activity, including T. asperellum 525, T. harzianum 610, and T. pseudokoningii 886. Here, we evaluated the effects of these 3 Trichoderma strains on preventing cucumber fusarium wilt through pot culture and greenhouse culture experiments. All 3 Trichoderma strains demonstrated higher control effects toward cucumber fusarium wilt than previous studies, with efficacies over 78%. Additionally, inoculation with the 3 Trichoderma strains significantly promoted the quality and yield of cucumbers. Among the 3 strains, Trichoderma 866 was the most effective, with disease control efficacy of 78.64% and a cucumber yield increase of 33%. Furthermore, seedlings inoculated with Trichoderma exhibited significantly increased measures of plant height, stem diameter, leaf area, aboveground fresh weight, underground fresh weight, chlorophyll content, and nitric nitrogen content, as well as the activities of several stress-resistance enzymes, including superoxide dismutase(SOD), peroxidase(POD), catalase(CAT), polyphenol oxidase(PPO), and ascorbate oxidase(AAO). In addition, the plants inoculated with Trichoderma showed decreased cell membrane permeability and malondialdehyde(MDA) content in the leaves. Together, our results suggest that T. asperellum 525, T. harzianum 610, and T. pseudokoningii 886 inoculations inhibit F. oxysporum infection, stimulate the metabolism in cucumbers, and enhance the activities of stress-resistance enzymes, which consequently promote the growth of cucumber plants, prevent cucumber fusarium wilt, and improve the yield and quality of cucumbers. T. harzianum is a commonly used biocontrol fungus, while few studies have focused on T. asperellum or T. koningense. In this study, strains of T. asperellum and T. pseudokoningii showed excellent plant disease prevention and growth promoting effects on cucumber, indicating that they also have great potential as biocontrol fungi.