我国蔬菜根结线虫发生、致害和绿色防控研究进展

根结线虫是威胁我国蔬菜生产的最重要病原物类群之一，寄主达3 000多种，每年造成的蔬菜损失价值高达5亿美元，目前缺乏安全高效的防治方法。近年来，随着保护地蔬菜种植面积的增加，根结线虫病对我国蔬菜生产的危害逐年加重，因此必须加强对蔬菜根结线虫病发生、致害和防治的深入研究。该文对近10来年我国根结线虫的发生分布和危害、主要种类、鉴定和快速诊断方法、成灾规律、与植物的互作及机理、绿色防控等方面的研究进展进行综述，并展望我国根结线虫的研究趋势。     

云南省紫茎泽兰根结线虫病病原种类鉴定

为明确云南省紫茎泽兰根结线虫病的病原种类,于2019年2月在云南省澜沧县林下三七种植区采集根部带有明显根结的紫茎泽兰根系进行根结线虫分离,通过观察所分离根结线虫的2龄幼虫、雌成虫、会阴花纹特征对其进行形态学鉴定,并利用序列比对、系统发育树分析、序列特异性扩增区段(sequence characterized amplified region,SCAR)对其进行分子生物学鉴定。结果表明,该病原线虫雌成虫会阴花纹呈圆形至卵圆形,背弓中等高或低平,侧区一侧或两侧延伸形成翼状,尾区有刻点,2龄幼虫、雌成虫形态特征及形态测量指标与北方根结线虫Meloidogyne hapla相似;该病原线虫rDNA的ITS序列和mtDNA的COI序列与NCBI数据库中已登录的北方根结线虫相应序列相似度较高,分别达99.35%和98.05%以上;该病原线虫rDNA的ITS序列、mtDNA的COI序列分别以99%、100%的支持率与北方根结线虫聚为同一分支;利用SCAR特异性引物,该病原线虫均能扩增出大小约1 500 bp的基因特异性条带。综合形态学和分子生物学鉴定结果将云南省紫茎泽兰根结线虫病病原种类鉴定为北方根结线虫。     

Soil solarization as a substitute for methyl bromide fumigation in greenhouse tomato production in Cyprus

Preplant soil fumigation with methyl bromide (MB) is presently standard practice in greenhouse tomato production. Since this compound is scheduled to be phased out by 2005, the possibility of using solarization as an alternative soil disinfestation method was examined in four greenhouse tomato trials. Solarization was applied for 8 weeks in July-August, using transparent polyethylene sheets for soil mulching, and compared with MB fumigation applied in September, before planting, at 80 g/m². Solarization raised the maximum soil temperature by 9°C and reduced the population density ofFusarium spp. in soil by 91–98%. Similar reductions of soil inoculum (95–99%) were obtained with MB fumigation. Both methods provided effective control of Fusarium wilt, Verticillium wilt and corky root rot on tomato plants. MB fumigation was in addition highly effective against root-knot nematodes, whereas nematode control with solarization did not exceed 50%. Both treatments resulted in similar fruit yield increases, ranging within 90–140% compared with plants grown in untreated soil. During the second cropping season following soil treatment, solarization exhibited two times higher residual effectiveness against vascular wilt diseases compared with MB fumigation. The latter treatment, however, was superior to solarization in its residual effectiveness against root-knot nematodes and to a lesser extent against corky root rot. Fruit yields from solarized and MB-fumigated soil during the second cropping season were higher than those obtained from untreated soil by approximately 35% and 60%, respectively. In Cyprus, solarization appears to be an effective alternative to MB fumigation in greenhouse tomato production, especially if integrated with other approaches enabling more effective nematode control.     

Effect of solarization and fumigant applications on soilborne pathogens and root-knot nematodes in greenhouse-grown tomato in Turkey

A study was conducted in two greenhouses with a history of Fusarium crown and root rot (Fusarium oxysporum f.sp.radicis-lycopersici, Forl) and root-knot nematodes (Meloidogyne javanica andM. incognita). During the 2005–06 growing season, the effectiveness of soil disinfestation by solarization in combination with low doses of metham-sodium (500, 750, 1000 and 1250l ha⁻¹) or dazomet (400 g ha⁻¹), was tested against soilborne pathogens and nematodes in an attempt to find a suitable alternative to methyl bromide, which is soon to be phased out. Solarization alone was not effective in the greenhouse with a high incidence ofForl. In the greenhouse with a low level ofForl, all the treatments tested reduced disease incidence, and were therefore considered to be applicable for soil disinfestation. In addition, root-knot nematode density decreased with all the treatments tested in both of the greenhouses.     

Induced Resistance to Cyst and Root-knot Nematodes in Cereals by DL-β-amino-n-butyric Acid

Foliar sprays and soil drenches with DL--amino-n-butyric acid (BABA) reduced the number of Heterodera avenae and H. latipons cysts on wheat and barley. Foliar sprays of wheat with 8000mgl^–1 BABA reduced the number of H. avenae cysts by 90%, whereas 2000mgl^–1 BABA was enough to reduce the number of H. latipons cysts by 79%. Multiple spray treatments with 2000mgl^–1 BABA at 10-day intervals reduced the number of H. avenae cysts on wheat and barley. A soil drench of wheat with 125mgl^–1 BABA reduced the number of H. latipons cysts by 93% and H. avenae cysts by 43%. Second-stage juveniles of these nematodes penetrated and formed syncytia in wheat roots soil-drenched with BABA. More adult males of H. avenae were produced in BABA (<250mg1^–1)-treated wheat roots (~76%) than in untreated roots (27%). Soil drenches with higher concentrations of BABA inhibited development of adult males and females. Several chemical elicitors of induced resistance were tested for their ability to reduce the number of H. avenae cysts on wheat. Only BABA was found to be an effective resistance inducer. The number of egg masses of an unidentified Meloidogyne sp. root-knot nematode, which infects only monocots, was also reduced by 95% by a soil drench of wheat with 500mgl^–1 BABA. Development of this nematode inside the BABA-treated roots was also inhibited.     

剂型、灌水量和土壤疏松度对甲维盐在土壤中分布及防治根结线虫效果的影响

为明确剂型、灌水量等对杀线虫剂防治根结线虫效果的影响,采用土壤薄层层析和高效液相色谱法分析了乳油和微囊悬浮剂型及不同灌水量对甲维盐在不同疏松度土壤中的分布特点,并以番茄为材料进行了根结线虫田间防治试验。结果表明,微囊悬浮剂型甲维盐随水迁移能力较乳油剂型强。药量相同、单株灌水量为1 600 m L时,2种剂型在疏松土壤中分布较为均匀,其次是800m L;200 m L和400 m L时,乳油剂型的分布均匀度明显大于微囊悬浮剂型。单株灌水量800 m L时2种剂型在致密土壤中分布的均匀度显著低于疏松土壤,乳油剂型略高于微囊悬浮剂型。番茄定植后单株甲维盐有效成分为10 mg、灌水量为800 m L和1 600 m L时,乳油剂型防效分别为67.90%和79.62%,微囊悬浮剂型分别为65.00%和77.50%,差异不显著。表明田间使用甲维盐乳油或微囊悬浮剂型防治蔬菜根结线虫时应尽量在移栽后随水冲施,且单株灌水量不低于800 m L。     

Nematicidal Activity of Chrysanthemum coronarium

Organic amendments and green manure are potential alternatives to the harmful chemical control means currently used against plant-parasitic nematodes. In this work, Chrysanthemum coronarium was applied to the soil as a green manure to control the root-knot nematodes Meloidogyne incognita and M. javanica. Chrysanthemum coronarium significantly reduced nematode infection of tomato roots and improved plant-top fresh weight, both in the greenhouse and in microplots. Other green manures, derived from Anthemis pseudocotula, wild chickpea (Cicer pinnatifidum), Geranium spp. and wheat, were not as effective as C. coronarium. Chrysanthemum coronarium, retained its nematicidal activity even when applied as a dried material. Only mature C. coronarium plants, in their flowering stage, exhibited nematode control activity, but the green plant parts were more effective than the flowers. An aqueous extract of C. coronarium exhibited in vitro, nematostatic activity towards M. incognita and M. javanica second-stage juveniles and inhibited their hatching from eggs and egg-masses; its nematostatic activity was expressed also against other phytonematode species such as Heterodera avenae and Pratylenchus mediterraneus, but did not affect the beneficial entomopathogenic nematode Steinernema feltiae.     

宁夏葡萄苗圃根结线虫病发病症状、为害程度及种类

为有效防治葡萄苗圃根结线虫病,采用拔根百分法和直接解剖法调查宁夏永宁县玉泉营镇葡萄苗圃葡萄苗木根结线虫病的发病症状、为害程度,并通过形态学和分子生物学对其种类进行鉴定。结果表明:15个葡萄品种的病害症状之间有一定差异。除抗砧3号外,其余14个葡萄品种均受根结线虫侵染,且不同葡萄品种受根结线虫为害的严重度差异明显,其中西拉、红芭拉蒂、爱神玫瑰、阳光玫瑰、马瑟兰、贝达和黑比诺上根结线虫病较严重,病情指数介于51.67～78.33之间,大青的病情指数最低,为21.67。不同葡萄品种的每根段雌成虫数量之间、每根段根结数量之间差异明显,其中葡萄品种贝达的每根段雌成虫数量和每根段根结数量最多,分别为12.47条和4.27个,葡萄品种黑比诺的每根段雌成虫数量和每根段根结数量较少,分别为3.60条和1.73个;每根段雌成虫数量随着每根段根结数量的增加而增加。根据根结线虫2龄幼虫、雌成虫和会阴花纹的形态学特征及分子生物学鉴定结果确定为害葡萄苗圃的根结线虫主要为南方根结线虫Meloidogyne incognita和花生根结线虫M. arenaria。除抗砧3号外,其它14个葡萄品种均不同程度的感染了南方根结线虫和花生根结线虫,且所有葡萄品种感染南方根结线虫的比例均较高。     

棉隆与淡紫拟青霉联合防治番茄根结线虫病的效果评价

为明确土壤熏蒸与生防微生物对番茄根结线虫病的联合作用效果,采用移栽前棉隆熏蒸处理后穴施淡紫拟青霉Paecilomyces lilacinus YES-2-14生防菌剂的方法在温室大棚开展了联合防治试验。结果表明,棉隆用量为35 g/m~2时,24周拉秧时线虫数量减少90%以上,防效达到85.5%;移栽时穴施浓度10~7~10~8CFU/g的淡紫拟青霉菌剂,24周拉秧时防效为44.4%~59.6%;而当线虫初始密度达到674头/100 g土时,单一菌剂防效只有12.5%,但番茄长势和产量显著提高。棉隆熏蒸后再联合施用淡紫拟青霉菌剂,2个月和5个月时防效比单一棉隆处理分别提高14.1%和21.8%,比单一菌剂处理提高14.3倍和4.1倍,产量较对照、棉隆熏蒸和单一菌剂处理分别提高22.1%、9.5%和2.4%,且该处理地块线虫数量始终最低,与对照差异显著,而淡紫拟青霉数量则高于单一淡紫拟青霉菌剂处理。表明棉隆和淡紫拟青霉菌剂联合使用不仅能显著降低线虫数量,有效防治番茄根结线虫病,还能够促进植株生长并提高产量。     

Evaluation ofAmaranthus sp. andVernonia amygdalina, and soil amendments with poultry manure for the management of root-knot nematodes on eggplant

The suppressive effect of vernonia (Vernonia amygdalina), amaranth (Amarathus sp.) and poultry manure on root-knot nematodes (RKNs) (Meloidogyne spp.) infecting eggplant (Solanum macrocarpon) was studied at two sites in southern Benin naturally infested with these nematodes. After 3 months, soil and root-inhabiting RKN populations were significantly less (P≦0.05) in the plots cropped with vernonia, amaranth, and eggplant amended with poultry manure (PM) at the rate of 40 t ha⁻¹ as compared with the rate of 20 t ha⁻¹ and with the control. Poultry manure was more effective after 2 months than after 3 months. Overall, vernonia was the most effective treatment affecting RKN populations in the roots and the soil. The use of these treatments in nematode management through rotation and co-planted crops is discussed. http://www.phytoparasitica.org posting August 6, 2008.     

Application of a putative fatty-acid binding protein to discriminate serologically the two European quarantine root-knot nematodes, Meloidogyne chitwoodi and M. fallax, from other Meloidogyne species

Two major proteins, Mcf-A67 and Mcf-B66, were identified by mini two-dimensional polyacrylamide gel electrophoresis in order to distinguish the two European quarantine root-knot nematodes, Meloidogyne chitwoodi and M. fallax, from eight other species. These quarantine proteinic markers have been microsequenced after enzymatic digestion. The internal amino acid sequences exhibit similarities to members of a family of low molecular weight intracellular lipid-binding proteins. Moreover, to explore a simple, rapid, and inexpensive way to identify the two quarantine nematodes, dot blot hybridizations were performed using an antiserum (A67) produced from the longest amino-acid sequence of the protein Mcf-A67. Although several proteins stained on the M. chitwoodi and M. fallax western blot membranes, the two nematodes were easily distinguished from other root-knot nematodes, on dot blot assays with soluble proteins extracted from a single female. Because of its specificity and sensitivity, the use of the A67 antiserum to improve the diagnosis of the two European quarantine root-knot nematodes is discussed.     

长枝木霉菌株TL16防治南方根结线虫的作用机理

为探明长枝木霉Trichoderma longibrachiatum菌株TL16防治南方根结线虫Meloidogyne incognita的作用机理,采用原生质体转化法获得绿色荧光蛋白(green fluorescent protein,GFP)标记菌株GFP-TL16,通过测定菌株TL16和GFP-TL16对南方根结线虫卵和2龄幼虫(2nd-stage juvenile,J2)的寄生与致死作用,其发酵液对卵孵化的抑制作用和对J2的致死作用,以及菌株GFP-TL16在黄瓜根系的定殖情况和菌株TL16对番茄根结形成的抑制作用来综合分析其作用机理。结果显示:菌株TL16菌丝对南方根结线虫卵无寄生作用,处理19 d后卵降解率为26.33%,致死作用较低;菌株TL16分生孢子悬浮液处理南方根结线虫J2后72 h的致死率为1.65%,且无寄生作用。菌株TL16发酵液处理南方根结线虫J2后48 h的校正死亡率为10.71%,处理卵15 d后对卵孵化的相对抑制率为77.11%。菌株GFP-TL16可定殖于黄瓜根系中,经菌株TL16处理后接种南方根结线虫J2,番茄根结减退率为55.88%。表明长枝木霉菌株TL16可通过抑制根结线虫卵孵化和诱导番茄产生抗病性来防治根结线虫病。     

Efficacy of a formulated product containing <Emphasis Type="Italic">Quillaja saponaria</Emphasis> plant extracts for the control of root-knot nematodes

The nematicidal effect of a formulated product containing extract from Quillaja saponaria was evaluated against the root-knot nematodes. The product QL Agri® 35 (QL) was tested to record the effect on second stage juveniles motility, egg hatch and also against field populations in greenhouse experiments contacted in three different locations of Greece. Convulsive movement of second stage juveniles of Meloidogyne incognita was recorded after exposure for 8 days at a series of doses, while the most paralyzed juveniles were counted at the dose of 8 mg l^?1. There was also a gradual decrease in the number of juveniles emerging from egg masses of the same nematode species when the dose of Q. saponaria was increased from 0 to 8 mg l^?1. In greenhouse experiments, the use of Q. saponaria could control root-knot nematodes and prevent nematodes increase in soil. The present study demonstrates that the use of Q. saponaria extract has the ability to control root-knot nematodes. Control given by Q. saponaria in field populations infecting cucumber was similar to that of cadusafos (Rugby®) and oxamyl (Vydate®) under the tested dosages and the specific conditions of the experiments.     

Selection of rhizobacteria able to produce metabolites active against Meloidogyne exigua

This work aimed to select potentially useful rhizobacteria for the control of Meloidogyne exigua, a widely disseminated root-knot nematode found in Brazilian coffee fields. Initially, crude metabolites produced in liquid medium by 69 isolated rhizobacteria strains (previously obtained from several plants collected in the south of Minas Gerais State, Brazil) were tested in vitro for lethality towards M. exigua second stage juveniles (J₂). Substances produced by strains of Bacillus cereus, B. megaterium, B. pumilus, B. thuringiensis, Enterobacter asburiae, E. cloacae and Paenibacillus macerans caused high mortality in J₂. Subsequently, the ten most active crude metabolites were selected and assessed for nematicidal activity in a biotest with M. exigua-inoculated coffee plants. Although strains of B. cereus, B. pumilus and P. macerans caused a significant reduction in the number of root galls and/or nematode eggs per plant 90 days after treatment, the best results were obtained with the application of B. megaterium strain 54-06.     

Evaluation of Natural Chemical Compounds Against Root-lesion and Root-knot Nematodes and Side-effects on the Infectivity of Arbuscular Mycorrhizal Fungi

The survival of two species of plant parasitic nematodes: the root-lesion nematode Pratylenchus brachyurus, and the root-knot nematode Meloidogyne javanica, was evaluated in saturated atmospheres of 12 natural chemical compounds. The infectivity of two isolates of arbuscular mycorrhizal fungi: Glomus mosseae and Glomus intraradices, under identical experimental conditions, was also determined. All the compounds tested exerted a highly significant control against M. javanica and among them, benzaldehyde, salicilaldehyde, borneol, p-anisaldehyde and cinnamaldehyde caused a mortality rate above 50% over P. brachyurus. The infectivity of G. intraradices was inhibited by cinnamaldehyde, salicilaldehyde, thymol, carvacrol, p-anisaldehyde, and benzaldehyde, while only cinnamaldehyde and thymol significantly inhibited mycorrhizal colonization by G. mosseae.     

Characterization of Rhizoctonia Species Associated with Foliar Necrosis and Leaf Scorch of Clonally-propagated Eucalyptus in Brazil

The objective was to identify and characterize the causal agent of foliar necrosis and leaf scorch of Eucalyptus spp. in Brazil. Nineteen putative isolates of Rhizoctonia obtained from Eucalyptus plants during clonal propagation were compared with isolates from other hosts and with tester strains of anastomosis groups of Rhizoctonia solani. Features compared were morphological characteristics of anamorphs and teleomorphs, numbers of nuclei per cell in the vegetative hyphae, anastomosis of hyphae, and ability to produce necrotic lesions on cuttings and damping-off of E. grandis×E. urophylla hybrid seedlings. Rhizoctonia solani AG1 (AG1-IB like) was the most frequent causal agent isolated from Eucalyptus plants and cuttings with symptoms of leaf scorch and foliar necrosis respectively. These isolates were highly virulent on Eucalyptus cuttings and presented naturally epiphytic growth on Eucalyptus shoots. Binucleate isolates and isolates of R. solani AG4 were also virulent on cuttings and were most virulent on Eucalyptus seedlings causing pre- and post-emergence damping-off. Virulence on Eucalyptus cuttings and seedlings was not restricted to a single species or anastomosis group of Rhizoctonia.     

Adhesive hyphae ofArthrobotrys oligospora: an ultrastructural study

Electron microscopic studies of the interaction of second-stage juveniles ofMeloidogyne hapla andArthrobotrys oligospora (CBS 289.82) strongly suggest that hyphae attachment to nematodes was mediated by a 0.1m thick layer matrix between the fungus and prey after contact with the nematode cuticle. The amorphous electron opaque matrix was irregularly distributed over the fungal surface, in some cases covering only the side attached to the nematode. Serial sections of adhesive hyphae showed that the extracellular matrix spread from the exact site of capture. A matrix with this thickness was never observed in the absence of nematodes.     

Suitability of weed species prevailing in Spanish vineyards as hosts for root-knot nematodes

Commercial vineyards in southern Spain were surveyed and sampled during October to December 2004 to determine the extent to which common weeds present were suitable hosts of root-knot nematodes infesting soils of those vineyards. Seven weed species commonly growing in grapevine soils in southern Spain were found infected by either Meloidogyne incognita or M. javanica: Amaranthus retroflexus (redroot pigweed), Anchusa azurea (ox-tongue), Chenopodium album (goosefoot), Erodium moschatum (musk stork’s bill), Malva rotundifolia (low mallow), Sinapis alba (white mustard), and Solanum nigrum (black nightshade). The host suitability of the weeds to root-knot nematodes was evaluated on the basis of root galling severity and nematode population densities in soil and roots. Also, the host–parasite relationship in these naturally Meloidogyne-infected weeds was examined. All the weed species in the study were considered suitable hosts for M. incognita and M. javanica because: (a) high Meloidogyne spp. populations occurred in roots and surrounding soil of the weed species; (b) the severity of root galling was high, and (c) well-established permanent feeding sites were observed in the histopathological studies of infected root tissues. In addition, this study presents the first reports of S. alba and A. azurea as hosts for M. incognita, and of E. moschatum as a new host for M. javanica, thus increasing the list of reported weed hosts for Meloidogyne spp. These results indicate that noticeable population densities of M. incognita and M. javanica can be maintained or increased in these weeds, at population levels higher than those previously reported for the same nematodes infecting grapevine roots. The weeds infesting vineyards thus represent an important source of inoculum of Meloidogyne spp., and furthermore may act as reservoirs of these nematodes which can be disseminated within or among vineyards by agricultural operations.     

Integrated Soybean Pest Management in Micro River Basins in Brazil

A Soybean IPM system was deployed in large, continuous river basin areas. The system was designed to improve the productivity of the rural environment by restoring balance among pests and their natural enemies. The system was developed with the active participation of various governmental, industrial and academic institutions, and it was based on the main approaches utilized by the EMBRAPA soybean IPM program referred here as IPM-Soybean.The work of IPM-Soybean in microbasins is being implemented in five counties in Paraná state (Campo Mourão, Mamborê, Missal, Cambé and Toledo) in an area of approximately 18,020ha of soybean involving 343 producers at various stages of implementation. The results obtained in the Rio do Campo basin in Campo Mourão, after four years of IPM-Soybean, showed drastic changes in pest control practices. Mean number of insecticide applications per cropping year in the river basin fell from 2.8 (1993/94 season) to 1.23 four seasons later. Biological control of the velvetbean caterpillar, Anticarsia gemmatalis, by Baculovirus anticarsia increased 57%, from 205ha treated with this biological product in the 1993/94 season to 2730ha in 1998. A mean of about 300,000 Trissolcus basalis (Wollaston) adults were released per year to control soybean stink bugs. As a consequence of the parasitoid release and the use of more selective insecticides, the number of insecticide applications to control stink bugs was significantly reduced. After two soybean seasons with IPM-Soybean in the river basin, mean applications for stink bugs decreased from 0.81 before the program started to 0.09 in 1996. The mean number of applications in the region and the state respectively were 9.20 and 11.44 times greater. In 1997 and 1998, the results again showed very low population densities of stink bugs in the river basin area, with a consequent reduction in insecticide applications for the control of these pests. Furthermore there was a substantial improvement in the range of products used in this area for soybean pest control. Broad spectrum products, used in 97.5% of the applications before start of IPM-Soybean (1993/94), were replaced with more selective products, especially biologicals and growth regulators. The latter had rarely been used by the river basin producers before the study (0.6% of the applications), but they accounted for 35.8% of the applications after four seasons with IPM-Soybean. IPM-Soybean developed in river basins has resulted in a more stable control system that, in turn, provides greater long-term equilibrium between the pests and the natural enemies populations in these soybean producing areas.     

Toxicity of fungal endophyte secondary metabolites to plant parasitic nematodes and soil-borne plant pathogenic fungi

Fungi isolated from the cortical tissue of surface sterilized tomato roots collected from field plots produced secondary metabolites in nutrition broth that were highly toxic toMeloidogyne incognita. Especially strains ofFusarium oxysporum were highly active with 13 of 15 strains producing culture filtrates toxic to nematodes. The mechanism of action of the toxic metabolites produced by the non-pathogenicF. oxysporum strain 162 with proven biological control ofM. incognita in pot experiments was investigated. These metabolites reducedM. incognita mobility within 10 min of exposure. After 60 min, 98% of juveniles were inactivated. Juveniles were initially inactivated within a few minutes of exposure, but with exposure of 5 h 50% of the juveniles were dead and 24 h exposure resulted in 100% mortality. In a bioassay with lettuce seedlings metabolite concentrations > 100 mg/l reduced the number ofM. incognita juveniles on the roots comparing to the water control. TheF. oxysporum toxins were highly effective towards sedentary parasites and less effective towards migratory endoparasites. Nonparasitic nematodes were not influenced at all. Metabolites of strain 162 also reduced significantly the growth ofPhytophthora cactorum, Pythium ultimum andRhizoctonia solani in vitro. Secondary metabolites of endophytic fungi on plant-parasitic nematodes and soil-borne fungi should be considered for control of plant parasitic nematodes and plant pathogenic fungi. The results also show the need for proper selection of target nematodes inin vitro bioassays.