Evaluation of bacterial biological control agents for control of root-knot nematode disease on tomato

Among 19 bacterial strains isolated in Yunnan from rhizosphere soils and plant tissues, Bacillus methylotrophicus strain R2-2 and Lysobacter antibioticus strain 13-6 exhibited the highest antagonistic activity against the tomato root-knot nematode Meloidogyne incognita in plate and greenhouse pot experiments. The two strains, when applied as soil drenches or seed treatments in greenhouse experiments, reduced root-knot severity and incidence on tomato compared to no-bacteria controls. In tomato field trials conducted in separate years, soil drench treatment with either strain reduced root-knot disease levels and increased yields compared to the control. Levels of disease control and yield enhancement provided by the strains were higher than those using the chemicals abamectin and carbofuran. This is the first report of B. methylotrophicus being used as a biocontrol agent against a plant parasitic nematode and the first demonstration that B. methylotrophicus and L. antibioticus can suppress disease caused by root-knot nematodes in the field.     

Evaluation of resistance to root-knot nematode (Meloidogyne incognita) in okra cultivars

The root-knot nematode, Meloidogyne incognita, is one of the major limiting factors affecting plant growth and yield causing an estimated $100 billion loss per year worldwide. Synthetic pesticides, though instantaneously effective, are usually prohibitively expensive, not readily available, may cause hazards to both man and livestock, and inflict injury to the environment. Notable among the alternatives to nematicides is the use of resistant cultivars which are inexpensive and eco-friendly. In the present studies, twelve okra (Abelmoschus esculentus L) cultivars were evaluated for their resistance against M. incognita under field conditions. Ten-day old okra plants of test cultivars were inoculated with 3000 freshly hatched second stage juveniles of M. incognita. The nematode caused reductions in various growth parameters of all the cultivars to varying levels over their respective controls. None of the cultivars was found completely resistant. The cultivar ‘Sharmeeli’ was highly susceptible as >100 galls were recorded on the roots. Sharmeeli also showed maximum reductions in growth among the cultivars evaluated. The cultivars Anmol and Sindha were susceptible with 71–100 galls. The cultivars Sabz Pari, Super Star, PMS-55 and PMS Beauty were moderately susceptible with 31–70 galls and comparatively less reductions in growth. Cultivars Sanam, Dikshah, Arka Anamika, Ikra-1 and Ikra-2 with 11–30 galls were rated as moderately resistant and showed less damage by the nematode as compared to susceptible cultivars and their planting could provide a useful tool to control root-knot nematodes.     

Enhanced defense responses of tomato plants against late blight pathogen Phytophthora infestans by pre-inoculation with rhizobacteria

The efficacy of resistance in tomato plants, induced by rhizobacteria isolated from Jeju Island in Korea, against late blight disease caused by Phytophthora infestans was tested. Among the bacterial isolates tested, pre-inoculation with four isolates induced an effective defense against late blight. The four isolates, TRH423-3, TRH427-2, KRJ502-1 and KRY505-3, were identified as Burkholderia gladioli, Miamiensis avidus, Acinetobacter quenomosp and Bacillus cereus, respectively, by sequencing the ribosomal intergenic spacer region. They also promoted the growth of tomato seedlings. To illustrate resistance mechanisms, the infection process by P. infestans was examined using a fluorescence microscope. There were no noticeable differences in the rate of germination and appressorium formation between the untreated and pre-inoculated plants. However, callose was more frequently formed at the penetration sites on the leaves of pre-inoculated plants than the untreated plants, suggesting that these rhizobacterial isolates induce defense responses against P. infestans.     

Biological control of the false root-knot nematode Nacobbus aberrans by Pseudomonas protegens under controlled conditions

The use of antagonistic biological agents, such as fungi and bacteria, offers an economical and safe strategy to manage plant–parasitic nematodes in infested fields. The false root-knot nematode, Nacobbus aberrans, is a damaging parasite of many agronomic and horticultural crops in South and North America. The management of this nematode is challenging and often not profitable for farmers. In greenhouse tests, conducted in Argentina, applications of the strain Pseudomonas protegens CHA0 and its isogenic derivative ARQ1 (used as control) at a rate of 10⁸ cfu ml⁻¹ suppressed infection and reproduction of N. aberrans on tomato roots. However, neither of the strains promoted plant root or shoot growth of the treated plants. Root colonization by the bacteria was assessed by specific PCR–RFLP protocols and fluorescence microscopy. The results obtained in this preliminary study were encouraging and showed the potential of P. protegens CHA0 to be used for the management of the false root-knot nematode on tomato.     

Induction of lignin and pathogenesis related proteins in dragon fruit plants in response to submicron chitosan dispersions

Chitosan is capable of triggering the induction of defence responses of the plants. This study was carried out to investigate in vitro and in vivo antifungal activity of submicron chitosan dispersions and also to determine the mechanism of induced resistance in dragon fruit plants by strengthening of cell wall and production of pathogenesis related proteins. Highest inhibition in radial mycelial growth (90.2%) was obtained by 600 nm droplet size at 1.0% chitosan concentration. None of the emulsifiers showed any antifungal effect during in vitro studies. Field studies showed the reduction of 22% disease incidence and 66% disease severity in anthracnose on dragon fruit plants treated with 600 nm droplet size as compared to control plants after one month of assessment. Lignin contents in cell wall of dragon fruit plants increased 2 folds in plants treated with 1.0% chitosan at 600 nm droplet size while 11 and 10 folds increase in β-1,3-glucanase and chitinase activity, respectively was also recorded in the plants inoculated with Colletotrichum gloeosporioides. The accumulation of lignin and higher level activity of pathogenesis related proteins in response to submicron chitosan dispersions may bolster in mounting effective resistance against the devastating disease of anthracnose.     

牛蒡寡糖诱导对小麦白粉病抗性的影响

为探讨小麦白粉病的防治新途径,采用不同浓度的牛蒡寡糖(BOS)(0.5～20.0 g/L)溶液处理小麦幼苗,分析其感染白粉病菌后病情指数、苯丙氨酸解氨酶(PAL) 、过氧化物酶(POD)和过氧化氢酶(CAT) 活性的变化.结果表明,随着BOS溶液浓度的升高,小麦幼苗的病情指数越来越低,对白粉病的抗性越来越强,在BOS浓度达到10.0 g/L时,抗性最好.BOS处理后的小麦幼苗叶片PAL、POD和CAT活性均高于对照,且在一定浓度范围内酶活性随着BOS浓度的增加而升高.PAL的活性在BOS处理浓度为5.0 g/L时达到最大,而POD和CAT活性在10.0 g/L时出现峰值.说明适宜浓度的牛蒡寡糖诱导可以提高小麦幼苗内部相关防御酶的活性,进而增强其白粉病抗性.     

外源信号物质诱导人参抗人参锈腐病的作用研究

探讨外源水杨酸(SA)茉莉酸甲酯(MeJA)诱导人参抗锈腐病的效果,以期通过施入外源物质来提高人参植株的抗性.测定了0～800mg·L-1不同浓度SA和MeJA溶液对人参锈腐病菌的影响,利用筛选的浓度(200 mg· L-1)处理二年生人参移栽苗,接种人参锈腐菌,调查人参锈腐病的发病率和病害严重度.结果表明,浓度为200 mg· L-1的SA和MeJA溶液对人参锈腐病菌无直接毒害作用,而经SA和MeJA处理的人参植株锈腐病发病率和病害严重度显著降低,SA的诱抗效果优于MeJA.     

Management of the root-knot nematode Meloidogyne graminicola infesting rice in the nursery and crop field by integrating seed priming and soil application treatments of pesticides

The study evaluates the effects of seed priming (5 g or ml kg⁻¹ seed) and soil application (2 kg or l ha⁻¹) of eight organophosphate pesticides on rice root-knot disease caused by Meloidogyne graminicola. Seed priming (SP) or soil application (SA) of phorate, carbofuran and chlorpyriphos (1000 J₂ of M. graminicola kg⁻¹ soil) suppressed galling in the rice nursery by 92 and 99%, 80 and 88% and 76 and 80%, respectively, over control. Relatively similar decreases in the galling were recorded when this nursery was grown for four months in the sterilized soils in earthen pots. Rice cv. PS-5 grown in naturally infested soil in earthen pots (1000 J₂ kg⁻¹ soil) became stunted, showing chlorotic foliage, and terminal galls developed on the roots. The treatment of SP + SA 15 + 30 days after planting (DAP) with phorate, carbosulfan, and chlorpyriphos significantly suppressed the root-knot development and improved the plant growth of rice over the controls (P ≤ 0.05). The overall effect of the SP + SA 15 DAP treatments was marginally weaker than the SP + SA 15 + 30 DAP treatments but statistically on par. Under field conditions, the greatest decrease in the galling occurred due to SP + SA 15 + 30 DAP of phorate (69−71%) and SP + SA 15 DAP (65−67%) followed by carbosulfan and chlorpyriphos. The yield of rice plants was also highest with phorate (32−36% and 29−34%) over the control during the two years of the study. The soil population of M. graminicola decreased by 58–84% over four months due to the phorate treatments. The study demonstrates that seed priming with phorate effectively controls nematode infections in the nursery and that soil application at 15 DAP (2 kg ai ha⁻¹) prevents root-knot development in an infested field under irrigated conditions. Use of SP + SA 15 DAP may enable to avoid one soil application of phorate in the field.     

Detecting resistance to the root-knot nematodesMeloidogyne hapla andM. chitwoodi in potato and wildSolanum spp.

Summary Various methods of screening for resistance to root-knot nematodes were compared and evaluated. Seedling populations ofSolanum spp., grown in clay pots and plastic tubes with silver sand and inoculated with juveniles ofMeloidogyne chitwoodi andM. hapla, showed large differences in the number of egg masses on roots 7 weeks after inoculation. The differences were reproducible when re-testing was done with cuttings and plants from tubers. No resistance toMeloidogyne spp. was observed with ten potato cultivars when grown in clay pots, plastic tobes or closed containers. Plants from tubers in growth pouches developed a large two-dimensional root system, and after inoculation with juveniles the infection process could be observed over 8 weeks. A method of infecting potato tuber tissue withMeloidogyne is described, using tuber slices in Petri dishes as a potential screening test for tuber resistance.     

Screening of three sweet potato (Ipomoea batatas L.) cultivars for resistance to different virulence groups of root-knot nematodes (Meloidogyne spp.) under controlled conditions

The susceptibility of three sweet potato cultivars (Ipomoea batatas L.) C4, TIS 3290 and TIS 9162 was evaluated against 156 isolates of Meloidogyne spp. with the aim to include resistant/tolerant sweet potato cultivars in a crop rotation scheme for the management of root-knot nematodes. The nematode isolates corresponded to races 1, 2 and 3 of Meloidogyne arenaria (n = 7), races 1, 2, 3 and 4 of M. incognita (n = 131) and Meloidogyne javanica (n = 18). Also, the isolates of M. incognita were differentiated in virulence groups: Pepper (n = 35), Pepper-Mi (n = 25), Tomato (n = 41) and Tomato-Mi (n = 30), depending on their ability to parasitize resistant pepper and tomato cultivars. The tested isolates of M. javanica parasitized C4 and TIS 3290, but not TIS 9162, whereas M. arenaria parasitized C4 and TIS 9162, but not TIS 3290, and M. incognita was able to parasitize the three sweet potato cultivars tested. C4 was the most susceptible cultivar to all nematode species tested, especially M. incognita, TIS 3290 was the most resistant and TIS 9162 was in between (7.2, 62.9 and 26.9% of resistant plants, respectively). Susceptibility of the sweet potato cultivars showed slight variations depending on the race or virulence group of M. incognita. The results suggest that sweet potato cultivars TIS 3290 and TIS 9162 may be used as rotation crops in fields where root-knot nematodes are present, their selection depending on the Meloidogyne isolates present. The use of resistant sweet potato cultivars would be preferably combined with other management practices to avoid virulence selection in nematode isolates.     

甲基茉莉酸、苯丙噻重氮、草酸 诱导油菜对菌核病的抗性

采用温室盆栽方法研究甲基茉莉酸(MeJA) 、苯丙噻重氮(BTH)和草酸(OA)诱导油菜对菌核病产生的 局部和系统抗性。结果表明, 0. 01～1. 0mmol/L BTH, 0. 1～1. 0mmol/L MeJA和0. 5～15mmol/L OA均能有效诱导 局部和系统抗性,系统抗性比局部抗性产生晚。Trypan blue染色结果表明,在MeJA,BTH和低浓度OA诱导过的油 菜叶片中,菌丝生长受到抑制,表现为菌丝变粗、弯曲。较低浓度(如0. 01mmol/L MeJA, 0. 5mmol/L OA)下虽然易 诱导局部抗性,而系统抗性较弱, 0. 01mmol/L MeJA几乎不诱导系统抗性。高浓度(如20mmol/L以上)OA有致病 作用。     

甘蓝型油菜抗倒性评价及抗倒性与株型结构的关系

以一批抗倒伏和易倒伏的甘蓝型油菜进行田间试验，研究甘蓝型油菜抗倒性评价指标及抗倒性与株型结构关系。结果表明，成熟期田间测定抗拉力可反映出品种的抗根倒能力，成熟期根重及根颈粗与品种的抗根倒能力也有密切关系。成熟期室内测定的茎秆抗折力矩可反映出品种抗茎倒能力，茎重，尤其是茎干重/茎秆长也能反映品种茎秆的抗倒能力。株高、茎秆重心高度/株高等形态指标与品种抗倒性有密切关系。株高、分枝点高/株高适中、重心高度/株高较小、分枝数适中、角果分布均匀、株型为紧凑型的品种较抗倒伏，单株生产力也较高，是理想的抗倒株型。     

稻瘟菌非致病性和弱致病性菌株对稻株诱导抗性的初步研究

稻瘟病是世界各稻区分布最广的病害之一。其病原菌Magnaporthe grisea (Hebert) Barr nov. comb. (anamorph Pyricularia oryzae Cavara)属高度变异的真菌。国内外曾对其复异进行了较长时期的研究,认为异核现象、准性生殖及非整倍性是变异的机制。近年来,     

Evaluation of spray programs containing famoxadone plus cymoxanil, acibenzolar-S-methyl, and Bacillus subtilis compared to copper sprays for management of bacterial spot on tomato

Bacterial spot caused by Xanthomonas euvesicatoria Jones et al. and Xanthomonas perforans Jones et al. is a major disease on fresh market commercial tomato in Florida. Fourteen field trials were conducted between 1999 and 2005 (10 in south Florida and four in north Florida) testing famoxadone plus cymoxanil (Tanos 50DF^®, E.I. du Pont de Nemours and Company, Wilmington, DE), Bacillus subtilis strain QST 713 (B. subtilis) (Serenade WPO^® or Serenade Max^®, AgraQuest, Inc., Davis, CA), and acibenzolar-S-methyl (ASM) (Actigard 50WG^®, Syngenta Crop Protection, Greensboro, NC) at different rates and in various application programs that were combined and rotated with copper hydroxide and mancozeb for management of bacterial spot. In field applied spray treatments containing famoxadone as a component, all of the programs significantly reduced bacterial spot severity on plants compared to the untreated control plants (UTC) and 97% of the programs were equal for disease suppression conferred by the copper-mancozeb standard. In spray programs containing ASM or B. subtilis plus copper hydroxide, treated plants had significantly reduced disease compared to the UTC plants and were not different from the plants treated with the copper-mancozeb standard. Yield data from small plots was unaffected. Several of the programs which used these compounds in rotation with copper-mancozeb provided similar levels of reduction in the disease severity for bacterial spot while reducing by 50% the amount of copper applied to plants. The effect of famoxadone plus cymoxanil on the survival of Xanthomonas in vitro did not cause a significant reduction in the bacterial population and was not determined to be directly bactericidal. However, greenhouse and field testing supports disease reduction of bacterial spot on plants treated with these compounds. The products tested in these trials may be useful, alternative tools for use in an integrated management program for bacterial spot on tomato.     

Induction of systemic resistance and defense-related enzymes after elicitation of resistance by rhizobacteria and silicon application against Ralstonia solanacearum in tomato (Solanum lycopersicum)

Bacterial wilt caused by Ralstonia solanacearum is a devastating plant disease affecting several economically important crops. To trigger plant-mediated systemic resistance against the pathogen, silicon and the rhizobacteria strain Bacillus pumilis were applied as elicitors. The impact of each elicitor in the induction of systemic resistance was evaluated by individual or dual application in pathogen inoculated tomato genotypes. Application of silicon and B. pumilis significantly reduced bacterial wilt incidence by 50.7 and 26.7%, respectively, in the King Kong 2 genotype (moderately resistant) and by 31.1 and 22.2%, respectively, in the L390 genotype (susceptible) but the dual application of the elicitors did not. The pathogen responsive lipoxygenase (LOX) activity was decreased in the silicon-treated tomato plants and no significant difference was detected in B. pumilis treated-pathogen inoculated plants compared to the control. In addition, a non-significant increases of peroxidase (PO) activity and phenylalanine ammonia lyase (PAL) activity were observed in either silicon/B. pumilis treated and pathogen inoculated plants. However, the PO, PAL and LOX activities declined during the dual application due to antagonistic effect of the two pathways triggered by the two elicitors. Therefore, separate application of each elicitor is recommended as the best strategy for induction of systemic resistance against bacterial wilt in tomato with silicon being a stronger inducer than B. pumilis.     

Effects of Pseudomonas fluorescens CHA0 on the Resistance of Wheat Seedling Roots to the Take-all Fungus Gaeumannomyces graminis var. tritici

Abstract

The aim of this study was to verify that wheat seedlings treated with Pseudomonas fluorescens CHA0 (CHA0 hereafter) before inoculation with Gaeumannomyces graminis var. tritici (Ggt), a pathogen of take-all, acquire induced resistance to Ggt. The soil with wheat seedlings growing on it was drenched with a suspension of CHA0 cells and inoculated with Ggt 24 h later. Then, the seedlings were grown in a glasshouse, and severity of take-all disease, fresh weights of root and shoot and lengths of root and shoot, and also the activities of soluble peroxidase (SPOX), ionically cell-wall-bound peroxidase (CWPOX), β-1,3-glucanase, β-1,4-glucanase and the concentration of total phenolic compounds in the root of the seedlings were examined. The results indicated that the treatment with CHA0 before inoculation with Ggt mitigated the disease severity significantly, and increased the root and shoot lengths and root and shoot fresh weights. The treatment with CHA0 increased the activities of SPOX, CWPOX, β-1-3-glucanase, β-1,4-glucanase and phenolic compounds in the wheat roots and the activities of SPOX and β-1,4-glucansee activities were highest at day 4 and those of CWPOX and β-1,3-glucanase at day 6 after inoculation with Ggt. The concentration of total phenolic compounds was also highest at day 6 after the inoculation with Ggt. The results suggest that the take-all suppressing effect of CHA0 may be related to enhanced defense response of the wheat roots.     

The control of isariopsis leaf spot and downy mildew in grapevine cv. Isabel with the essential oil of lemon grass and the activity of defensive enzymes in response to the essential oil

The aim of this study was to evaluate the potential of the essential oil of lemon grass (Cymbopogon citratus) for the control of isariopsis leaf spot (Pseudocercospora vitis) and downy mildew (Plasmopara viticola) in grapevine cv. Isabel, the effect of the essential oil on the productivity of the grapevines and the effect of the essential oil on the activity of the enzymes chitinase and catalase. The experiment was conducted in a commercial vineyard over two consecutive crop cycles. Each plant in the experiment was subjected to one of the following nine treatments: 0, 0.5, 1.0, 2.0 or 4.0 mL L⁻¹ essential oil, Tween^® 80%, Bordeaux mixture, Acibenzolar-S-methyl and mancozeb. An analysis of the area under the disease progress curve showed a quadratic response by both diseases to the doses of essential oil during the first and second crop cycles. The essential oil treatments also increased the number and mass of the clusters of fruit as well as the productivity and desirable chemical characteristics of the grape. The activity of chitinase increased as a result of the essential oil treatments, whereas the activity of catalase decreased. The essential oil at doses of 1.0 and 2.0 mL L⁻¹ can serve as an alternative means of controlling isariopsis leaf spot and downy mildew and can also serve to improve the fruit quality of grapes cv. Isabel in tropical regions.     

Multiple-fungicide resistance to carbendazim,diethofencarb, procymidone,and pyrimethanil in field isolates of Botrytis cinerea from tomato in Henan Province,China

Botrytis cinerea, the fungal pathogen responsible for causing gray mold in tomatoes, frequently exhibits multiple fungicide resistance. In this study, the sensitivity to carbendazim (Car), diethofencarb (Die), procymidone (Prc), and pyrimethanil (Pyr) was determined for 263 isolates of B. cinerea from diseased fruit and leaves of tomato plants from 87 commercial greenhouses in Henan Province of China during 2013 and 2014. Of the isolates tested, 89%, 93%, 85%, and 86% were resistant to Car, Die, Prc, and Pyr, respectively. Twelve phenotypes were found having different sensitivity patterns, and multi-fungicide resistance was found to one, two, three or four fungicides. The percentage of multiple fungicide resistant isolates (Car^RDie^RPrc^RPyr^R) was 68%. Mycelial growth, spore production, and mycelial dry weight were not significantly different between resistant and sensitive phenotypes of the B. cinerea isolates. However, the virulence of Car^SDie^RPrc^SPyr^S isolates was higher than that of the other fungicide-resistance phenotypes. All of the carbendazim and diethofencarb resistant phenotypes carried a point mutation at codon 198 (E198A or E198K) or 200 (F200Y) in the β-tubulin gene, and all of the procymidone resistant phenotypes carried point mutations at codons 369 (Q369P) and 373 (N373S) in the Bos1 gene. These results provided important reference data for assessment of resistance risk of B. cinerea isolates from commercial tomato transplants in Henan Province, indicating the existence of a high frequency of resistance to carbendazim, diethofencarb, procymidone, and pyrimethanil revealing the necessity of a more integrated control of gray mold.     

内生解淀粉芽孢杆菌TB2菌株活性物质诱导辣椒果抗疫病的生化机理

利用内生解淀粉芽孢杆菌TB2菌株的活性物质喷施辣椒果后0、24、48、72 h接种辣椒疫霉病菌进行防治辣椒疫病试验,结果表明,TB2菌株的活性物质对辣椒果疫病的发生有一定控制作用,活性物质处理间隔24 h后再接种病菌,其防病效果得到显著提高。生化机制研究表明,TB2的活性物质处理可使辣椒果的可溶性蛋白含量、丙二(MDA)含量、超氧阴离子自由基(O.-2)产生速率及过氧化物酶(POD)、超氧化物歧化酶(SOD)活性发生显著变化。病菌处理的辣椒果,可溶性蛋白含量、MDA含量和O.-2产生速率及POD、SOD、过氧化氢酶(CAT)、苯丙氨酸氨解酶(PAL)活性均会显著升高,且其最大值高于活性物质处理果的相应最大值。活性物质和病菌共同处理的辣椒果,POD和PAL的活性可上升到高于病菌单独处理的峰值,但MDA含量和O.-2产生速率及SOD和CAT活性均较低,与对照的接近,表现出相对稳定状态。由此推断,TB2的活性物质诱导辣椒果抗疫病与POD、PAL等酶的活性相关。     

The application of an excised root assay for the determination of susceptibility or resistance to root-knot nematodes (Meloidogyne spp. Goeldi) in potatoes (Solanum spp. L.)

Excised roots of six potato selections were challenged with egg inoculum of fourMeloidogyne species:M. chitwoodi, M. javanica, M. incognita, andM. hapla. The dual cultures were established under aseptic conditions in 60 × 15 mm petri plates on a defined agar medium. Egg hatch occurred within one week and galling within four weeks at incubation temperature of 24°C. All six potato selections, Russet Burbank, White Rose, PAS 3065,S. sparsipilum 365343.A19-12,S. sparsipilum ×S. phureja 856D68-1, and NDTX8731-1R supported reproduction of all fourMdoidogyne species. Corresponding greenhouse tests were run with eachin vitro experiment to establish the efficacy of this screening technique compared to greenhouse methods. Results from the greenhouse assays had less variation within cultivar treatment than did the results from thein vitro assays. Thein vitro assay, however, may be conducted with less labor, time, and greenhouse facility management.