A New Root-Knot Nematode, Meloidogyne baetica n. sp. (Nematoda: Heteroderidae), Parasitizing Wild Olive in Southern Spain

ABSTRACT High infection rates of wild olive (Olea europaea sp. sylvestris) feeder roots and soil infestation by a new root-knot nematode were found in sandy soil at Vejer de la Frontera (Cádiz), southern Spain. Morphometric traits and analyses of the nematode esterase electrophoretic pattern as well as of the internal transcribed spacer 1 (ITS1)-5.8S gene and D2-D3 fragment of the 28S gene of rDNA showed that specimens differed clearly from known root-knot nematodes. Studies of host-parasite relationships showed a typical susceptible reaction in naturally infected wild olive plants and in olive planting stocks (cvs. Arbequina and Picual) artificially inoculated with the nematode. However, the nematode did not reproduce in artificially inoculated chickpea, pea, and tomato. Because of the ability of this new nematode to infect wild and cultivated olives only, we suggest the common name, "Mediterranean olive root-knot nematode." The species is herein described and illustrated, and named as Meloidogyne baetica n. sp. The new root-knot nematode can be distinguished from other Meloidogyne spp. by (i) the perineal pattern, which is almost similar to that of M. artiellia, characterized by distinct inner striae forming two distinct longitudinal bands, extending throughout the perineum to just below the vulva; (ii) female excretory pore anterior to the level of stylet knobs, excretory pore distance from anterior end/length of stylet ratio extremely small (0.5 to 0.8); and (iii) second-stage juveniles with elongate-conoid tail. Phylogenetic trees derived from maximum parsimony analyses showed that M. baetica is closely related to M. artiellia, the cereal and legume root-knot nematode.     

Effect of root-knot nematodes on cajanol accumulation in the vascular tissues of pigeonpea after stem inoculation with Fusarium udum

The effect of root-knot nematodes ( Meloidogyne javanica and M. incognita ) on production of the isoflavonoid phytolexin, cajanol, was investigated in pigeonpea ( Cajanus cajan ) plants infected with Fusarium udum , the causal organism of Fusarium wilt. Seven-day-old seedlings of a wilt-resistant pigeonpea cultivar, ICP 9145 and a wilt-susceptible cultivar, Malawi Local, both of which were moderately susceptible to the nematode, were grown in soil infested with 2000 Meloidogyne juyeniles per plant. A duplicate set of plants remained free from nematodes. Twenty-one days later, all the plants were inoculated with F. udum by stem puncture. Quantitative estimates of cajanol in the vascular tissues were made at intervals up to 15 days after inoculation with the fungus. No external symptoms of wilt appeared in any plants of the wilt-resistant cultivar in the absence of the nematode. However, when inoculated with the nematode, two thirds of the plants developed wilt symptoms. Cajanol levels were lower in both the wilt-resistant and wilt-susceptible plants in the presence of the nematode than in its absence, although this effect was considerably more marked in the wilt-resistant cultivar. These results indicate that the root-knot nematode is capable of breaking resistance of ICP 9145 to Fusarium wilt and that at least part of the mechanism of this effect is retarded cajanol accumulation.     

海南省蔬菜根结线虫发生种类与分布

2014年-2018年, 对海南省蔬菜根结线虫病害进行了田间随机采样调查和病原种类分子鉴定?结果显示, 蔬菜根结线虫病在海南18市县均有发生, 且大部分旱田连作地块病株率达到80%以上?进一步对采集的295份根结线虫样本种类进行了分子鉴定, 共鉴定出象耳豆根结线虫?南方根结线虫和爪哇根结线虫3种病原种?其中, 象耳豆根结线虫单一检出率达到62.37%, 南方根结线虫单一检出率为23.39%, 爪哇根结线虫的检出率仅为5.76%, 象耳豆根结线虫和南方根结线虫复合侵染检出率为8.47%?除五指山市样本以外, 海南其余17市县样本均检测到象耳豆根结线虫侵染为害?本研究显示象耳豆根结线虫为海南省蔬菜上的优势病原根结线虫种类, 该结果对指导品种布局?制定根结线虫病害的防治策略具有重要意义?     

西南部分地区荞麦根结线虫种类与地理分布

近年来,随着我国荞麦面积的迅速增加,根结线虫病呈逐年加重趋势,严重影响了荞麦的产量与品质。本研究于2014年对西南10个地区17个乡镇秋播苦荞麦根结线虫的分布、发生种类以及危害程度进行了调查。结果表明,西南地区危害荞麦的根结线虫种类有南方根结线虫(Meloidogyne incognita)、爪哇根结线虫(M.javanica)和花生根结线虫(M.arenaria)3种,其中南方根结线虫为优势种群。田间根结线虫种群大多数为单一种群,23.5%的样品为南方根结线虫与爪哇根结线虫(或花生根结线虫)组成的混合种群。调查发现前作为烟草或马铃薯的地块,荞麦根结线虫发生危害严重,平均被害株率为6.0%~77.5%,病情指数为1.4~26.1,而前作为玉米的荞麦根结线虫则较轻,被害株率最高为16.0%,相应病情指数为2.3。     

Control of root-knot nematodes (Meloidogyne spp.) in Hungary1

Under Hungarian climatic conditions, root-knot nematodes, and other phytoparasitic nematodes, generally cause damage in certain plant growing areas. These pests are present in glasshouses throughout the country. In the field the nematodes occur in sandy soil in the vegetable-growing areas of southern and central Hungary. The following species of root-knot nematodes have so far been reported to occur in Hungary: Meloidogyne incognita, M. arenaria, M. hapla, M. javanica, M. incognita acrita, M. thamesi and M. naasi. There are 10 specific nematicides registered for nematode control in the country. These chemicals are mainly applied in glasshouses where their use produces most economic return. Nematoderesistant cultivars are only available in the case of tomato. The nematodes cause the heaviest damage on glasshouse cucumber.     

Histological Characterization of Resistance to Different Root-Knot Nematode Species Related to Phenolics Accumulation in Capsicum annuum

ABSTRACT In the pepper Capsicum annuum CM334, which is used by breeders as a source of resistance to Phytophthora spp. and potyviruses, a resistance gene entirely suppresses reproduction of the root-knot nematode (Meloidogyne spp.). The current study compared the histological responses of this resistant line and a susceptible cultivar to infection with the three most damaging root-knot nematodes: M. arenaria, M. incognita, or M. javanica. Resistance of CM334 to root-knot nematodes was associated with unidentified factors that limited nematode penetration and with post-penetration biochemical responses, including the hypersensitive response, which apparently blocked nematode migration and thereby prevented juvenile development and reproduction. High-performance liquid chromatography analysis suggested that phenolic compounds, especially chlorogenic acid, may be involved in CM334 resistance. The response to infection in the resistant line varied with root-knot nematode species and was correlated with nematode behavior and pathogenicity in the susceptible cultivar: nematode species that quickly reached the vascular cylinder and initiated feeding sites in the susceptible cultivar were quickly recognized in CM334 and stopped in the epidermis or cortex. After comparing our data with those from other resistant pepper lines, we suggest that timing of the resistance response and the mechanism of resistance vary with plant genotype, resistance gene, and root-knot nematode species.     

植物源提取物防治根结线虫的分子研究

针对内生真菌的生物防治,重点阐述了中草药提取物的杀虫活性、植物抗线虫基因的发现以及植物防御系统激发的最新研究.植物源提取物作为潜在的线虫杀虫剂,为生物源农药开发和利用提供新的思路.同时,对于今后线虫杀虫剂开发中存在的问题及解决方案进行了探讨,就其发展方向提出了建议.     

Meloidogyne virulence locus molecular marker for characterization of selected mi-virulent populations of Meloidogyne spp. is correlated with several genera of betaproteobacteria

Resistance to root-knot nematodes in tomato is conferred by the Mi resistance gene to the three most important species of Meloidogyne: M. arenaria, M. incognita, and M. javanica. Nevertheless, the Mi gene is unable to inhibit the reproduction of selected and naturally Mi-virulent populations of root-knot nematodes. As pathogenicity assays are time consuming, molecular markers were developed for the easy identification of Mi-virulent populations of Meloidogyne. The sequence characterized amplified region-Meloidogyne virulence locus (MVC) molecular marker is reported to differentiate Mi-avirulent and naturally Mi-virulent from selected Mi-virulent populations. This marker was used to compare acquired virulence in populations of M. javanica from Spain. The original populations used to develop the MVC marker were included as control for reference. Results showed that this marker did not amplify genomic DNA extracted from single juveniles or females of any of the populations tested either from Spain or Japan. In silico analyses performed with the recently published complete genome of M. incognita, indicated that the MVC marker is not correlated to a MVC or to any eukaryotic organism but to several betaproteobacteria genus from the family Comamonadaceae.     

海南岛葫芦科蔬菜根结线虫危害性调查与种类鉴定初报

对海南11个市县的葫芦科蔬菜进行根结线虫危害性调查,结果显示,大部分地区发病率在50%以上;其中琼中腰子、澄迈长安和琼中营根主产菜区的病情指数分别达到了47.12、47.14和40,已造成严重损失。采自海南11个县市5种葫芦科蔬菜24个根结线虫种群经单卵块纯化培养,运用形态学和同工酶的方法对其进行种类鉴定,结果表明,南方根结线虫占总数的79.17%,爪哇根结线虫占总数的12.5%,根结线虫疑似新种或新纪录种占总数的8.3%。南方根结线虫是危害海南葫芦科蔬菜的优势种。     

辽宁省保护地蔬菜根结线虫发生规律及防治对策*

2008-2009年,对辽宁省蔬菜主产区保护地蔬菜根结线虫病的危害程度进行调查,以辽宁省的辽中、辽西、辽南、辽东4个地区的黄瓜大棚为对象,对棚内土壤中根结线虫的发生规律进行系统研究。结果表明：辽宁省保护地蔬菜根结线虫病为南方根结线虫危害。该病在辽宁省保护地蔬菜上发生已相当普遍,其中以连作7年以上的黄瓜、番茄、苦瓜易感寄主为主的老菜棚的发病率达到100%,病情指数达到80以上。4个地区大棚土壤中根结线虫数量普遍随土层的加深呈明显减少的趋势;并且根结线虫数量普遍随着大棚前边缘向内的距离加大呈明显增加的趋势。结合辽宁省根结线虫发病特点,提出相应的防治措施。     

A new root-knot nematode, Meloidogyne silvestris n. sp. (Nematoda: Meloidogynidae), parasitizing European holly in northern Spain

High infection rates of European holly ( Ilex aquifolium ) feeder roots by an unknown root-knot nematode were found in a holly forest at Arévalo de la Sierra (Soria province) in northern Spain. Holly trees infected by the root-knot nematode showed some decline and low growth. Infected feeder roots were distorted and showed numerous root galls of large (8–10 mm) to moderate (2–3 mm) size. Morphometry, esterase and malate dehydrogenase electrophoretic phenotypes and phylogenetic trees of sequences within the ribosomal DNA (rDNA) demonstrated that this nematode species differs clearly from other previously described root-knot nematodes. Studies of host-parasite relationships showed a typical susceptible reaction in naturally infected European holly plants, but did not reproduce on a number of cultivated plants, including tomato, grapevine, princess-tree and olive. The species is described here, illustrated and named as Meloidogyne silvestris n. sp. The new root-knot nematode can be morphologically distinguished from other Meloidogyne spp. by: (i) roundish perineal pattern, dorsal arch low, with fine, sinuous cuticle striae, lateral fields faintly visible; (ii) female excretory pore level with stylet knobs, or just anterior to them, EP/ST ratio about 0·8; (iii) second-stage juveniles with hemizonid located 1 to 2 annuli anterior to excretory pore and short, sub-digitate tail; and (iv) males with lateral fields composed of four incisures, with areolated outer bands. Phylogenetic trees derived from maximum parsimony analysis based on 18S, ITS1-5·8S-ITS2 and D2–D3 of 28S rDNA showed that M. silvestris n. sp. can be differentiated from all described root-knot nematode species, and it is clearly separated from other species with resemblance in morphology, such as M. ardenensis , M. dunensis and M. lusitanica .     

Solarization of soil in piles for the control of Meloidogyne incognita in olive nurseries in southern Spain

The potential of solarization to control Meloidogyne incognita in piles of soil used at olive nurseries in southern Spain was studied in 1999 and 2000. Kaolin and soil infested with free eggs and egg masses of the nematode in nylon bags were buried 20 and 40 cm deep inside conical piles of soil 80 cm high and with a base diameter of 1 m. Soil piles were solarized for 3 weeks in July and August. The effect of various periods of solarization was assessed by egg hatch bioassays in sterile water, and by infectivity to tomato plants. Maximum soil temperature at 20 cm depth in solarized piles was 47·4°C in 1999 and 48·2°C in 2000, compared with 32·9°C and 31·7°C in nonsolarized piles. Solarization reduced egg hatch by > 95% compared with nonsolarized samples, irrespective of type, burial depth and location of inocula in a soil pile. Egg hatch of egg mass-infested samples buried at 20 cm depth was higher than that of free eggs buried at the same depth. The differential effect associated with burial depth and type of inoculum was not found in solarized piles. In nonsolarized piles, hatch of free eggs from samples buried at 40 cm depth was higher than that from samples buried at 20 cm depth. Egg hatch in samples from solarized piles was lower than that from nonsolarized piles. A bioassay of tomato plants in 2000 confirmed the reduction in infectivity of free eggs buried in solarized soil piles. Under the conditions in southern Spain, solarization of 40 cm-high piles of soil for 3 weeks can therefore be used for the control of root-knot nematodes in potting soil for olive nursery production.     

我国南方地区主要根结线虫DNA变异的RAPD分析

 本试验用5组随机引物对来自我国南方地区的30个根结线虫种群进行RAPD分析,并从中筛选出多态性较好的引物12个。共扩增出179条DNA多态带,各供试种群间存在着丰富的遗传多态性。扩增结果表现出种间差异大于种内差异的共同趋势,这表明上述12个引物能够较客观地反映种群间亲缘关系的远近。北方根结线虫与另外3种线虫(南方根结线虫、爪哇根结线虫、花生根结线虫)的亲缘关系最远;在3种主要根结线虫中,爪哇根结线虫与南方根结线虫的亲缘关系相对较近。基于种群间的相似系数分析和应用UPGMA法构建的聚类树状图,显示出不同的根结线虫在较低的相似性系数范围聚类,而绝大多数种内的不同种群均以较高的相似性系数聚在一起,这与形态分类基本一致,反映了形态学分类的分子遗传本质,同时也表明了应用RAPD技术进行根结线虫亲缘关系分析和种类鉴定具有合理性和可行性。本文还对RAPD方法对南方根结线虫小种鉴定的可能性进行了初步探讨。     

甘肃省白银市保护地蔬菜根结线虫病病原鉴定

2012年5月-7月在甘肃省白银市采集了番茄、黄瓜、豇豆等几种保护地蔬菜的25个根结线虫群体.直接解剖获得雌虫,通过雌虫会阴花纹观察,结合酯酶和苹果酸脱氢酶(MDH-EST)测定,鉴定出所收集的25个群体中有16个为南方根结线虫[Meloidogyne incognita (Kofoid&White)],9个为爪哇根结线虫[M.javanica (Treub)].南方根结线虫为优势种群,本次试验也是首次在甘肃省发现爪哇根结线虫.     

真菌Sr18代谢产物对根结线虫胁迫下黄瓜叶片保护酶的影响

为探究根结线虫胁迫下丝状真菌Sr18代谢产物对黄瓜的作用机理,采用温室盆栽及人工接种试验,研究了不同浓度的Sr18代谢产物对南方根结线虫胁迫下黄瓜叶片保护酶的影响。结果表明,线虫侵染黄瓜根部以后,黄瓜叶片SOD、POD和CAT活性减弱,PPO和PAL浓度降低。施加不同浓度的Sr18代谢产物,能够使线虫胁迫下的黄瓜叶片SOD、POD和CAT活性增强,使PPO和PAL的含量增加,说明Sr18代谢产物能够提高黄瓜的保护酶活性与含量,增强黄瓜对南方根结线虫的抗性。     

生防链霉菌JH108-2的发酵培养基优化

 为了提高链霉菌菌株JH108-2的发酵滤液对植物寄生线虫的毒力,采用正交试验设计,优化其发酵培养基。使用优化培养基[大豆粉20g、(NH₄)₂SO₄ 6g、蛋白胨3g、葡萄糖40g、酵母粉6g,NaCl 2g、K₂HPO₄ 0.6g.CaCO₃ 2g、蒸馏水1000mL、pH 6.0]所得的发酵滤液,稀释10倍,处理南方根结线虫(Meloidogyne incognita)二龄幼虫,24h后校正死亡率达95.4%,比对照培养基的发酵滤液所产生的校正死亡率高6.9%,差异达到极显著水平(P=0.01)。     

AM真菌和根结线虫互作对黄瓜生长及生理特征的影响

通过盆栽试验研究了丛枝菌根真菌Arbuscularmycorrhiza（AM）和南方根结线虫Meloidogyneincognita互作对黄瓜幼苗生长及生理特征的影响。结果表明,预先接种地表球囊霉Glomusversi—forme再接种线虫处理的鲜、干重较只接种线虫的处理显著增加;预先接种AM真菌再接种线虫处理的根结线虫病情指数、AM真菌侵染率较只接种线虫的处理明显降低;接种线虫使黄瓜植株的过氧化物酶（POD）、过氧化氢酶（CAT）及苯丙氨酸解氨酶（PAL）、总酚含量有不同程度提高,其中预先接种根内球囊霉G．intraradices对黄瓜植株起到了较好的保护作用;与对照相比,只接种线虫处理的叶片磷含量降低了25．26％,预先接种AM真菌再接种线虫处理的叶片磷含量较只接种根结线虫的含量高。     

Local and Systemic Induced Resistance to the Root-Knot Nematode in Tomato by DL-beta-Amino-n-Butyric Acid

ABSTRACT Chemical inducers of pathogenesis-related proteins and plant resistance were applied to tomato plants, with the aim of inducing resistance to the root-knot nematode Meloidogyne javanica. Relative to control plants, foliar spray and soil-drenching with dl-beta-amino-n-butyric acid (BABA) reduced root-galling 7 days after inoculation, as well as the number of eggs 30 days after inoculation. Other chemicals (alpha- and gamma-amino-n-butyric acid, jasmonic acid, methyl jasmonate, and salicylic acid) were either phytotoxic to tomato plants or did not improve control of root-knot nematodes. Fewer second-stage juveniles invaded BABA-treated tomato roots, and root-galling indices were lower than in control tomato plants. Resistance phenomena in seedlings lasted at least 5 days after spraying with BABA. Nematodes invading the roots of BABA-treated seedlings induced small, vacuolate giant cells. Postinfection treatment of tomato plants with BABA inhibited nematode development. It is speculated that after BABA application tomato roots become less attractive to root-knot nematodes, physically harder to invade, or some substance(s) inhibiting nematode or nematode feeding-site development is produced in roots.     

番茄抗性品种与黄瓜轮作对根结线虫的防治作用

通过室内盆栽试验探讨对根结线虫具有不同抗性的番茄品种与黄瓜轮作减轻根结线虫病危害的可行性。试验结果表明:抗性品种多菲亚能极大降低土壤中根结线虫2龄幼虫和卵的密度,减轻黄瓜根结线虫病的危害;抗性砧木Beaufort也能显著地降低土中根结线虫和卵的密度,但下茬黄瓜根结线虫的危害与对照相比没有显著差异。表明抗性品种与感病品种轮作能有效地防止根结线虫的发生。     

Physiological Variation in the Genus Meloidogyne as Determined by Differential Hosts

Four species of root-knot nematodes, namely Meloidogyne incognita (Kofoid et White) Chitwood, M. javanica (Treub) Chitwood, M. arenaria (Neal) Chitwood, and M. hapla Chitwood have a wide host-range and are widely distributed throughout the world. For each of the species there are certain non-hosts and these are different for each species, thus making it possible, through the use of differential hosts, to identify the species as well as detect pathogenic variations within a species. A knowledge of the nature and extent of this variation is needed for the development of varieties which possess a broad base for resistance.