Differential expression of a β-1,4-endoglucanase induced by diet change in the foliar nematode Aphelenchoides fragariae

We identified and characterized a β-1,4-endoglucanase, Afr-ENG-1, in the foliar nematode Aphelenchoides fragariae that is differentially expressed when the nematode feeds on fungi or plants. When individuals from hosta plants were transferred to a fungus culture, expression of the enzyme decreased 1,812-fold after five generations on the fungus diet. Afr-eng-1 was readily detected in the genome of 75% of nematodes from the plant population but only in 38% of the diet-changed population. The gene cannot be detected in nematodes maintained on fungus for over 100 generations. Diet was also associated with changes in nematode body size and in the severity of symptoms caused on hosta leaves. Plant-diet nematodes caused larger lesions and were longer and thinner than fungus-diet nematodes. Nematodes moved from a plant diet to a fungus diet for five generations had the same body size as the nematodes that had fed on the fungus for 100 generations. Full-length sequences of Afr-eng-1 were obtained and found to encode a glycosyl hydrolase family 5 protein. This is the first β-1,4-endoglucanase and plant-parasitism-related gene described in the genus Aphelenchoides.     

Systemic nematicidal activity of fluensulfone against the root-knot nematode Meloidogyne incognita on pepper

BACKGROUND: Fluensulfone, a new nematicide of the fluoroalkenyl group, has proved to be very effective in controlling root‐knot nematodes, Meloidogyne spp., by soil application. The systemic activity of this compound against M. incognita on peppers via soil drenching and foliar spray was evaluated. RESULTS: Root application of fluensulfone via soil drenching showed slight and no nematode control activity when applied 4 and 10 days, respectively, after inoculation. A single foliar spray of peppers with a fluensulfone solution at 3.0 g L^?1 prior to inoculation reduced the galling index by 80% and the number of nematode eggs by 73–82% of controls. The reduction in these parameters by fluensulfone was much higher than that obtained with oxamyl or fenamiphos at the same concentration. This activity was also observed when the plants were sprayed 21 days before inoculation. A series of experiments suggested that foliar spray with fluensulfone prior to inoculation reduces nematode invasion. However, foliar spray after inoculation did not inhibit nematode development inside roots. CONCLUSION: Fluensulfone showed relatively high nematode control activity when sprayed on the foliage before inoculation. Fluensulfone may be used as a foliar application, in addition to soil application, for root‐knot nematode control. Copyright © 2011 Society of Chemical Industry     

Evaluation of chemical and integrated strategies as alternatives to methyl bromide for the control of root-knot nematodes in Greece

Current environmental awareness has led to a greater demand for alternative nematode control strategies. Three field experiments were established to compare management tactics on cucumber in commercial greenhouses naturally infested with root-knot nematodes (Meloidogyne spp). Cucumber rootstocks which have shown resistance to soil-borne diseases were tested to reveal any resistance/tolerance to root-knot nematodes, and integration of these rootstocks with nematicides was investigated. Metham-sodium and 1,3-dichloropropene (1,3-D) provided good control of nematode populations when their application was followed by the application of a non-fumigant nematicide such as cadusafos or oxamyl. Neither fumigant could provide season-long control of nematode populations, and a further application of cadusafos was required for satisfactory control. The efficacy of metham-sodium was significantly increased when injected into soil in comparison with its application through the drip irrigation system. The use of rootstocks resistant to soil-borne fungal pathogens used together with chemical means of nematode control provided promising results for their further use in integrated strategies as alternatives to methyl bromide. However, the latter was the superior treatment for the control of root-knot nematodes in soil infested with residues of galled roots. Dazomet, metham-sodium nor the non-fumigant nematicides oxamyl and fenamiphos could reduce nematode population as efficiently as methyl bromide. None of the chemicals tested except methyl bromide could enter galled roots and kill surviving nematodes.     

鸢尾种苗中截获的草莓滑刃线虫形态和分子鉴定

草莓滑刃线虫是一种重要的叶和芽寄生线虫,可侵染多种重要花卉、经济作物,被列入我国检疫性有害生物名录。目前,草莓滑刃线虫的鉴定以形态学鉴定为主,但一线口岸常分离不到雌虫,鉴定困难。本研究采用形态特征和分子特征相结合的方法对草莓滑刃线虫进行鉴定:对鸢尾种苗中截获的滑刃属线虫群体雌虫、雄虫进行形态观察和形态测计并与已报道文献进行比较;PCR扩增该群体的rDNA-ITS区并测序,构建基于ITS序列的系统发育进化树;采用草莓滑刃线虫特异引物对该群体DNA样本进行扩增。结果表明:该群体与草莓滑刃线虫俄罗斯种群的测量值相接近;系统进化关系分析显示,该群体与NCBI公布的草莓滑刃线虫均处于同一进化支;采用草莓滑刃线虫特异引物PCR扩增可产生片段大小为169 bp的目的条带。     

Control ofDitylenchus dipsaci in garlic by bulb and soil treatments

Garlic bulbs heavily infected withDitylenchus dipsaci were chemically treated to control the nematode. Treatment with ethoprop resulted in greatest reduction of nematodes early in the season; oxamyl provided good control; methomyl was ineffective. Treating the soil with aldicarb or phenamiphos was also effective. Later in the season, nematode populations increased with all treatments and crop damage was severe. In a soil heavily infested with this pathogen, nematode populations in the untreated plots were initially low but had increased rapidly 100 days after planting, resulting in total loss of plants by the end of the season. Excellent control was obtained by treating the soil with methyl bromide (MB) or solar heating with transparent polyethylene sheets prior to planting, although MB treatment caused severe stunting of the plants. Treatment with ethylene dibromide (EDB) controlled the pathogen initially, but later in the season the population level increased. The yields (kg/m²) were: untreated, 0; solar heating, 2.325; EDB, 0.813; and MB, 1.152.     

广西香蕉植物线虫病发生情况调查

作者于2009-2010年,对广西香蕉线虫病发生情况进行了实地调查,分别采集根际土壤和根系样本共253份,调查结果表明：广西香蕉主要有害线虫有根结线虫、矮化线虫、短体线虫、肾状线虫、螺旋线虫、滑刃线虫、纽带线虫、垫刃线虫和小环线虫,其中根结线虫发生最普遍也最严重,根结病指最高达60,土壤根结线虫虫口密度最高达307头/100 mL土。〖JP〗     

辽宁省水稻品种种子携带干尖线虫的调查

对辽宁省主要水稻品种的种子上携带水稻干尖线虫(Aphelenchoides besseyi)的群体数量进行了调查,结果表明沈稻3、辽粳294、旱禾、沈农389、1437、辽盐188、辽盐34等26个品种的种子上携带有水稻干尖线虫。胚乳发育正常的饱粒种子中带虫率、线虫虫量最高,空粒种子带虫率最低。     

41.7%氟吡菌酰胺悬浮剂不同施药时期对黄瓜根结线虫的防治效果

根结线虫在内蒙古设施黄瓜生产中平均造成30%以上的产量损失,危害严重时导致绝收,为了明确41.7%氟吡菌酰胺SC有效防治黄瓜根结线虫的最佳施药时期,在定植后当天、3 d、7 d和15 d分别使用41.7%氟吡菌酰胺SC稀释1 500倍液灌根;以定植后当天使用对照药剂20%噻唑膦EW稀释1 000倍液灌根、10%噻唑膦GR 2 kg/667 m2地面撒施及未施药处理为对照,施药71 d后41.7%氟吡菌酰胺SC处理区的土壤根结线虫虫口减退率分别为43.75%、37.39%、32.05%、18.12%,对黄瓜根结线虫的防治效果分别为66.4%、60.8%、55.6%、33.4%,对照药剂处理根结线虫虫口减退率分别为36.4%、25.0%,防治效果分别为64.6%、39.8%。研究结果表明,〖JP+1〗41.7%氟吡菌酰胺SC稀释1 500倍药液于设施黄瓜定植后当天和3 d灌根处理对根结线虫的防治效果较好,都达到60%以上,增产明显,分别为25.1%和39.2%,本研究为41.7%氟吡菌酰胺SC防治黄瓜根结线虫提供理论依据。     

Influence of a Fungus-Feeding Nematode on Growth and Biocontrol Efficacy of Trichoderma harzianum

ABSTRACT A fungivorous nematode, Aphelenchoides sp., was isolated from field soil by baiting with mycelium of the biocontrol fungus Trichoderma harzianum ThzID1, and subsequently was maintained on agar cultures of the fungus. Interactions between the nematode and the green fluorescent protein-producing transformant, T. harzianum ThzID1-M3, were investigated in both heat-treated (80 degrees C, 30 min) and untreated field soil. ThzID1-M3 was identified in soil by epifluorescence microscopy. When ThzID1-M3 was added to soil as an alginate pellet formulation, addition of the nematode (10 per gram of soil) significantly reduced radial growth and recoverable populations of the fungus, and the effect was greater in heat-treated soil than in untreated soil. Addition of ThzID1-M3 to soil pretreated with the nematode (10 per gram of soil) stimulated nematode population growth for approximately 10 to 20 days, whereas nematode populations decreased in the absence of added Trichoderma sp. When sclerotia of Sclerotinia sclerotiorum were added to soil (10 per 200 g of soil) with ThzID1-M3 (40 pellets per 200 g of soil), addition of Aphe-lenchoides sp. (2,000 per 200 g of soil) reduced the number of sclerotia colonized by ThzID1-M3. These results suggest that fungivorous nematodes may be a significant biotic constraint on activity of biocontrol fungi in the field.     

Differential effect of nematicide treatments on tuber yields in early- and late-maturing potato cultivars

A 3-year field study examined the effects of 1,3-dichloropropene or oxamyl on tuber yields in four early- and five late-maturing potato cultivars. The nematicide treatments increased total tuber yield by ≈ 12% for early-maturing cultivars, but by less than 2% for the late-maturing cultivars. The treatments reduced the numbers of root lesion nematodes (primarily Pratylenchus penetrans ), which were the dominant plant parasitic nematodes at the site. Covariance analysis indicated that treatment effects on total tuber yield were no longer significant after adjusting for root lesion nematodes in soil at harvest, confirming that these nematodes had a direct effect on the host. As expected, there were significant inverse relationships ( P  = 0·05) between total tuber yields and numbers of root lesion nematodes in soil and in roots. However, the application of nematicides in late-maturing cultivars was found to be of no practical value. Crop production systems designed to reduce or optimize pesticide applications can form one criterion in the development of more environmentally sensitive management strategies. The merit of utilizing chemicals for root lesion nematode control are considered with regard to cultivar maturity.     

玉米矮化病株及其根际土壤内线虫的分离与分子鉴定

为探究玉米矮化病的病原,采用Ludox TM悬浮法对从吉林和辽宁采集的玉米矮化病株及其根际土壤和健康株根际土壤分离的线虫进行了鉴定和种群比较,并对矮化病株茎基组织PCR产物进行测序分析。结果表明:玉米健康株根际土壤分离出29属线虫,特有线虫是角咽线虫属Actinolaimus和木盾移线虫属Peltamigratus,矮化玉米病株根际土壤分离出28属线虫,特有线虫为锥线虫属Dolichorus,二者相同的线虫属有27个。矮化玉米病株根际土壤中植物线虫种类和数量明显比健康株根际土壤中高,线虫种群数量差异明显的主要是矮化线虫属Tylenchorhynchus、短体线虫属Pratylenchus和刺线虫属Belonolaimus,病株根际土壤中这3个属线虫总数约占病株土壤分离线虫总数的20.23%、11.27%和10.40%。玉米矮化病株茎基组织中短体线虫属和矮化线虫属数量占优势,2个属的线虫数量分别占测序总数的22%、14%(吉林长岭)和16%、20%(辽宁黑山)。表明玉米矮化病的发生与植物线虫相关,很可能是多种线虫共同引起的病害。     

应用斯氏线虫防治桑皱鞘叶甲

应用斯氏属小卷蛾线虫Steinernema carpocapsae(A24品系）对桑皱鞘叶甲Abirus fortunei(Baly）进行了室内和田间感染试验。在25～30℃的室内条件下，1500～2000条线虫/ml的处理使桑皱鞘叶甲幼虫感染率达86%～100%。在9～10月份，先清除田间杂草，然后在桑园开沟，按45～75万条线虫/m^2的剂量施用小卷蛾线虫，对幼虫的防治效果可达88.3%～96.2%，并使次年成虫的出土数量减少46.2%～86.7%。此种防治方法，避免了在春蚕期间摘叶饲蚕与桑皱鞘叶甲化学防治的矛盾，显示出应用昆虫病原线虫防治桑皱鞘叶甲的良好前景。     

4种药剂对豇豆根结线虫病的防治效果

为了更好地防治豇豆根结线虫病,对4种杀线虫剂进行了田间防效评价试验。结果表明,供试的4种药剂均对豇豆根结线虫有一定的防效,能控制土壤中２龄幼虫的数量和抑制根结的形成,其中10％噻唑膦颗粒剂对豇豆根结线虫的防效最好,具有低残留、持效期长等优点,可作为防治豇豆根结线虫的首选药剂; 其次为５％阿维菌素颗粒剂。同时供试的4种药剂在试验剂量范围内均对豇豆安全,建议在生产上推广应用10％噻唑膦颗粒剂和５％阿维菌素颗粒剂来防治豇豆根结线虫病。     

Rice nematode problems in Nigeria: Their occurrence,distribution and pathogenesis

Abstract

In a survey of plant parasitic nematodes associated with or affecting rice throughout Nigeria, some important nematode pests, especially the white tip disease nematode, Aphelenchoides besseyi and the rice root nematodes Hirschmanniella spinicaudata and H. oryzae were identified from seed, soil and root samples from swamp rice fields respectively. The sugarcane cyst nematode, Heterodera sacchari occurred in swamp rice fields only around the major sugarcane estates in Nigeria. The root‐knot Meloidogyne incognita and the root lesion nematode, Pratylenchus brachyurus were also encountered in upland (rainfed) rice fields. The white tip disease nematode, A. besseyi occurred at low levels in soils and rice seeds throughout the country. High population levels of H. spinicaudata and H. oryzae were encountered especially in areas where monoculture of rice is practised. General chlorosis, poor tillering and significantly reduced yield have been observed due to H. spinicaudata. Rice plants attacked by H. sacchari also showed intense chlorosis, delayed and reduced tillering and reduced grain yield. The roots of attacked plants were twiggy, very necrotic and blackened. The root‐knot M. incognita and the root lesion nematodes P. brachyurus have both been observed to reduce rice yields. Rice cultivars screened for reactions to the nematodes showed varying degrees of susceptibilities. Some varieties were however resistant to the root‐knot nematode, M. incognita.     

Comparison of sandy soils suppressive or conducive to ectoparasitic nematode damage on sugarcane

ABSTRACT Two South African sandy soils, one suppressive and the other conducive to ectoparasitic nematode damage on monoculture sugarcane, were compared. Analysis of field transects indicated that the suppressive soil displayed a comparatively higher population of the weak ectoparasite Helicotylenchus dihystera, whose predominance among ectoparasitic nematodes is known to limit yield loss caused by more virulent phytonematodes. Soil type was identical at both sites (entisols), but the suppressive soil had a higher organic matter content and a lower pH, which correlated with H. dihystera population data. In contrast, microclimatic differences between the two field sites were unlikely to be responsible for the suppressive or conducive status of the soils, as shown in a greenhouse experiment. The two soils exhibited a bacterial community of the same size but with different genetic structures, as indicated by automated ribosomal intergenic spacer analysis (RISA). The number of culturable fluorescent pseudomonads was higher for the conducive soil, probably because extensive root damage caused by ectoparasitic nematodes favored proliferation of these bacteria. This study shows that apparently small differences in soil composition between fields located in the same climatic area and managed similarly can translate into contrasted nematode communities, ectoparasitic nematode damage levels, and sugarcane yields.     

Effects of Potato-cyst Nematodes (Globodera pallida) and Soil pH on Root Growth, Nutrient Uptake and Crop Growth of Potato

Potato-cyst nematodes (Globodera pallida) cause severe yield losses in potato. Plants infected with potato-cyst nematodes generally have reduced concentrations of nitrogen, phosphorus and potassium in the foliage. This study investigated whether reduced growth of nematode-infected potato is caused by nutrient limitation.Experiments in the field and in containers showed that phosphorus concentration correlated best with total crop biomass at early stages of growth. The role of phosphorus in nematode damage was further investigated in the field and in the Wageningen Rhizolab. The experimental field was infested with potato-cyst nematodes and two levels of nematode density were established by fumigation with a nematicide. Prior applications of calcium carbonate resulted in pHKCl levels of 4.8 and 6.1. Two levels of phosphorus fertiliser were applied: either 0 or 225kgPha–1. In the Wageningen Rhizolab, soil of both pH levels from the field was used after treatment with 1MRad gamma irradiation to kill the nematodes. Subsequently, half of the soil was inoculated with cysts to give a nematode density of 30 viable juveniles per gram of soil.In the field, nine weeks after planting, the total crop biomass ranged from 107gm–2 for the treatment with nematodes at pHKCl 6.1 without phosphorus fertiliser to 289gm–2 for the fumigated treatment at pHKCl 4.8 with phosphorus fertiliser. The differences in total biomass for the various treatments were explained by differences in foliar phosphorus concentration. Nematodes induced or aggravated P deficiency and reduced total biomass. This was not the only damage mechanism as at high, non-limiting levels of foliar phosphorus concentration, nematodes still reduced total biomass.In the Wageningen Rhizolab, directly after planting, the number of roots visible against minirhizotrons was reduced by nematodes. However, the increase of root number in the nematode treatment continued longer than in the control, until root number was higher than that of the control. The compensary root growth of the nematode treatment was restricted to the top 30cm and nematodes reduced rooting depth.High soil pH reduced growth, mainly by reducing the availability of phosphate. Both nematodes and high soil pH reduced nutrient uptake per unit root length. Our results lead us to suggest an interaction between nematodes and soil pH, with nematode damage being higher at pHKCl 6.1 than at pHKCl 4.8.     

佛手山药线虫病害发生调查与病原鉴定

湖北省武穴市佛手山药是“农产品地理标志产品”, 近年佛手山药线虫病害严重, 为此在病源地采集山药土样进行了病害发生调查及病原鉴定?通过形态特征观察?形态特征测计值比较, 以及基于rDNA 28S D2/D3区序列构建短体线虫属线虫群体系统进化树并进行SCAR验证, 对引起佛手山药线虫病的病原线虫进行种类鉴定?结果表明:危害佛手山药的病原线虫的群体特征与咖啡短体线虫Pratylenchus coffeae相似?系统进化树显示:该病原群体与中国香港群体(HQ688677.1)聚类在一个分支上, 置信度达99%?因此, 将该群体线虫鉴定为咖啡短体线虫Pratylenchus coffeae?此外田间调查表明:线虫主要集中分布于10~20 cm深度的土层中, 线虫密度随土壤深度的增加而呈递减趋势, 且连作1年地较连作2年地的线虫数量少, 施用阿维菌素的地块较未施用的线虫数量少?上述结果表明, 在佛手山药生产中实行轮作和科学选择药剂防治, 避免连作, 可降低该线虫发生量?     

进口百合种球携入植物线虫数量风险初探

对荷兰和新西兰进口的8种百合种球入境时所携带的植物线虫种类和数量进行了检测,并进行了灭菌土栽培,以监测和探讨携入的线虫数量在花卉一个生长周期后的变化,进而为评价携入的植物线虫的数量风险提供科学依据。研究结果表明栽培前种球内的优势植物线虫——拟滑刃属和滑刃属线虫数量在百合一个生长周期结束后均以几何级数的速率增长;栽培前检出数量极少的垫刃属线虫,生长周期结束后却大量出现;栽培前少量检出的短体属、矮化属、根结属、丝尾垫刃属和伪垫刃属的线虫,周期结束后,均未再检测到。本研究结果初步表明拟滑刃属、滑刃属和垫刃属线虫具有较高的潜在危险性,其他检出的植物线虫类群潜在风险较小。     

Evaluating Pochonia chlamydosporia in a double-cropping system of lettuce and tomato in plastic houses infested with Meloidogyne javanica

The effect of Pochonia chlamydosporia , a facultative fungal parasite of nematode eggs, alone or in combination with oxamyl was evaluated in a double-cropping system of lettuce and tomato in unheated plastic houses infested with Meloidogyne javanica at two sites for two consecutive growing seasons. An additional treatment of methyl bromide fumigation was included to compare crop yield in nematode-free vs. nematode-infested soil. Final population densities, reproductive rate, root gall rating, and egg production were determined after each crop. Pochonia chlamydosporia was isolated from nematode eggs up to nine months after application to soil. The fungus survived in the rhizosphere for the entire growing season at one site, but only at low densities. Final population densities of M. javanica decreased after cultivation of lettuce and increased after tomato, and this pattern of population fluctuation was unaffected by treatment, experiment or site. The reproductive rate on lettuce was equal to or below 1, and it was similar among treatments in both experiments at both sites. Eggs were not found on lettuce roots. On tomato, the reproductive rate in the fungus + oxamyl treatment was significantly lower ( P  < 0·05) than other treatments in experiment 1 at both sites. Fungus + oxamyl consistently reduced root gall ratings on tomato in all cases, but numbers of eggs per g root varied depending on treatment. Methyl bromide-treated plots remained free of M. javanica at the end of the 2-year study.     

Chemical alternatives to methyl bromide for the control of root-knot nematodes in greenhouses

The complete phase-out of methyl bromide from use in developed countries by 1 January 2005 will cause many problems in agricultural industries that are now heavily reliant on its use. Three field experiments were established to compare management tactics on tomato and cucumber in commercial greenhouses naturally infested with root-knot nematodes (Meloidogyne spp). Reduction of nematode juveniles in soil and roots to nil detection levels was observed in all plots following soil fumigation with methyl bromide. A significant reduction of nematode juveniles and root-galling index was observed in plots treated with metham-sodium, dazomet and 1,3-dichloropropene compared with the control and plots treated with non-fumigant nematicides. Reduction of the nematode population led to an increase in fruit yield. However, data collected from the second cultivation season indicated that single control methods such as fumigant or contact nematicides alone cannot drastically decrease initial nematode population and those nematodes which escape control lead to population increase by the end of the cropping season.