New sources of resistance to Meloidogyne incognita race 3 in wild cotton accessions and histological characterization of the defence mechanisms

Accessions of Gossypium spp., some of them never previously tested, were evaluated for resistance to a local isolate of Meloidogyne incognita race 3 in greenhouse experiments. Nematode infection was characterized based on the galling and egg mass indexes and the reproduction factors (RF). Root‐knot nematode reproduction among the newly tested accessions varied from highly susceptible (AS0188, AS0189) to moderately resistant (MT123 no. 3), and some accessions showed highly reduced nematode reproduction (CIR1343, CIR1348, Fai Mui). Histological observations of two resistant accessions (G. barbadense CIR1348 and G. hirsutum TX‐25, respectively) showed that resistance occurs through a two‐stage mechanism in the first accession and through a single‐stage mechanism in the second. Parasitism is blocked early after second‐stage juvenile (J2) penetration or during its initial tissue migration (CIR1348) and the development of later‐stage juveniles into female adults is suppressed at a later stage (TX‐25 and CIR1348). Fluorescence and bright light microscopy showed that root cells surrounding nematodes exhibit a hypersensitivity‐like reaction, with the accumulation of presumably phenolic compounds and the presence of necrotic cells that limit the development of nematodes and the formation of giant cells. Underdeveloped giant cells with degenerated cytoplasmic content were found in small numbers in CIR1348 and in large numbers in TX‐25, along with deformed nematodes. The full characterization of the defence mechanisms of novel sources of resistance to the root‐knot nematode in cotton constitutes a first step towards their use in crop improvement.     

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.     

Cropping systems for the management of phytonematodes

Damage caused by nematodes is one of the limiting factors in crop production. Traditional nematode management is based on the use of crop rotations, resistant cultivars, nematicides, or combinations of these methods. For a crop like peanut (Arachis hypogaea), cultivars resistant to root-knot nematodes are not available. There are soybean (Glycine max) cultivars resistant to some of the species of root-knot nematodes (Meloidogyne spp.); however, most fields have nematode infestations composed of mixtures of species. Research at Auburn has shown that tropical crops can be used effectively in rotation to manage nematode problems. Rotations with American jointvetch (Aeschynomene americana), castor (Ricinus communis), hairy indigo (Indigofera hirsuta), partridge pea (Cassia fasciculata), sesame (Sesamum indicum), and velvetbean (Mucuna deeringiana) have resulted in good nematode control and increased yields of peanut and soybean. Some crops (castor, sesame) are considered ‘active’ in that they produce compounds that are nematicidal, whereas others (e.g. corn, sorghum) are simply non-host, that is, ‘passive’.     

Root‐knot nematodes do not affect black root rot disease severity on watermelon and bottle gourd

The effects of root‐knot nematodes on black root rot of watermelon and bottle gourd were studied using field surveys and co‐inoculation tests with Meloidogyne incognita (southern root‐knot nematode) and Diaporthe sclerotioides. The results of the field survey suggested that root‐knot nematodes had little effect on either the severity of black root rot or infection with D. sclerotioides. None of the three fields showed a significant positive correlation between disease severity and nematode gall index, with low correlation coefficients. Co‐inoculation experiments under controlled conditions found no significant effect of root‐knot nematodes on black root rot of watermelon and bottle gourd based on area under disease progress curves (AUDPC). These results were supported by the quantities of DNA of the two agents in root tissues because no significant difference was found between dual‐ and single‐inoculation treatments with M. incognita and/or D. sclerotioides. These findings suggest that root‐knot nematodes probably do not affect the infection of watermelon or bottle gourd roots by D. sclerotioides or the incidence of black root rot in these crops caused by this fungus.     

Host suitability of Vitis rootstocks to root‐knot nematodes (Meloidogyne spp.) and the dagger nematode Xiphinema index,and plant damage caused by infections

The host suitability of commercial Vitis rootstocks commonly used in Spain (161‐49C, 41B, 1103P, 110R, 140Ru and SO4) to root‐knot nematodes (Meloidogyne arenaria, M. incognita, M. javanica) and Xiphinema index, and damage caused by nematode infection were determined under controlled conditions. The three root‐knot nematodes reproduced with a rate higher than one in all rootstocks, indicating that they are suitable hosts for these nematodes. Growth of rootstocks infected with the root‐knot nematodes was less vigorous than that of nematode‐uninfected controls in the majority of the rootstocks studied. Root infection resulted in moderate to severe root galling in all rootstocks. The shoot and main stem diameters appeared to be the most sensitive variables of damage caused by infection by Meloidogyne spp., with reduction rates from 36% and 53% in 161‐49C to 57% and 66% in 140Ru, respectively. The shoot height was not significantly affected by the root‐knot nematodes and the root fresh weight generally increased as a consequence of intensive galling. The nematode X. index caused significant root damage with a reproduction factor higher than one in all rootstocks. However, reproduction factor was significantly influenced by the rootstock and significantly decreased by about 12‐fold (5·7 to 18·1‐fold) with the increase in inoculum density from 100 to 1000 nematodes per plant. The root dry weight was reduced by X. index infections, and was the plant growth variable most affected by the nematode infection in all rootstocks at both inoculum densities. Meloidogyne arenaria, M. incognita, M. javanica and X. index, prevalent in many world vineyards, are all shown to have a damaging effect on the six tested rootstocks.     

Alteration of gene expression profile in the roots of wild diploid Arachis duranensis inoculated with Ralstonia solanacearum

Bacterial wilt caused by Ralstonia solanacearum is a serious disease of peanut (Arachis hypogaea) in China. However, the molecular basis of peanut resistance to R. solanacearum is poorly understood. Arachis duranensis, a wild diploid species of the genus Arachis, has been proven to be resistant to bacterial wilt, and thus holds valuable potential for understanding the mechanism of resistance to bacterial wilt and genetic improvement of peanut disease resistance. Here, suppression subtractive hybridization (SSH) and macroarray hybridization were employed to detect differentially expressed genes (DEGs) in the roots of A. duranensis after R. solanacearum inoculation. A total of 317 unique genes were obtained, 265 of which had homologues and functional annotations. KEGG analysis revealed that a large proportion of these unigenes are mainly involved in the biosynthesis of phytoalexins, particularly in the biosynthetic pathways of terpenoids and flavonoids. Subsequent real‐time polymerase chain reaction (PCR) analysis showed that the terpenoid and flavonoid synthesis‐related genes showed higher expression levels in a resistant genotype of A. duranensis than in a susceptible genotype, indicating that the terpenoids and flavonoids probably played a fundamental role in the resistance of A. duranensis to R. solanacearum. This study provides an overview of the gene expression profile in the roots of wild Arachis species in response to R. solanacearum infection. Moreover, the related candidate genes are also valuable for the further study of the molecular mechanisms of resistance to R. solanacearum.     

The development of juveniles of Heterodera schachtii in roots of resistant and susceptible genotypes of Sinapis alba,Brassica napus,Raphanus sativus and hybrids

The development ofHeterodera schachtii Schm. (beet cyst nematode, BCN) juveniles in roots of resistant and susceptible genotypes belonging to cruciferous crop species and hybrids was studied from 4 to 28 days after inoculation. No difference in root penetration by larvae was observed between resistant and susceptible plants.The development of nematodes in roots from resistant plants ofRaphanus sativus L., resistant xBrassicoraphanus Sageret and a resistant hybrid xBrassicoraphanus x Brassica napus L. was similar. BCN resistance in these three sources of plant material appeared to be related to an increased male:female nematode ratio as compared to the ratio found in susceptibleR. sativus plants.Also in resistant plants ofSinapis alba L. and a resistant intergeneric hybridS. alba x B. napus the increase in male:female nematode ratio, as compared to the ratio found for susceptibleS. alba cultivars and a susceptible intergeneric hybridS. alba x B. napus, seemed to be related with the observed resistance. In roots of the resistantS. alba and of a resistant hybridS. alba x B. napus, however, BCN resistance might also be due to a slower development of larvae and increased necrosis of root cells at the site of larval penetration.     

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     

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。     

酸黄瓜南方根结线虫病抗性的解剖学及其细胞学研究

为探明酸黄瓜南方根结线虫病抗性的内在机理,以抗病材料酸黄瓜与感病材料北京截头为试材,对南方根结线虫侵入与幼虫发育、根的解剖学及线虫侵入诱导形成的取食位点的细胞学结构进行了比较研究。结果表明,抗病材料酸黄瓜根系中线虫侵入率极显著低于感病材料北京截头(P0.01),酸黄瓜能够有效抵抗南方根结线虫的侵入;抗、感材料根系中雌雄虫个体比例分别为1∶12和1∶5(P0.01),抗性反应能够抑制线虫取食和幼虫发育;酸黄瓜根的解剖结构与普通栽培黄瓜相似,酸黄瓜的根不具有结构抗性;抗病反应中细胞发生过敏性坏死,感病反应中无此类现象发生。抗侵入、抑制取食和坏死反应是抗病材料酸黄瓜抗性反应的主要特征。     

和田鞘氨醇杆菌AMCC 100218菌株的筛选、鉴定及杀线虫特性

为筛选对南方根结线虫具有致死效果的生防细菌,从山东省10个县市蔬菜主产区番茄根际土壤中分离可培养细菌,采用离体杀线虫试验测定分离菌株对南方根结线虫Meloidogyne incognita的致死活性,结合生理生化特征及分子生物学方法对高效杀线虫菌株进行分类鉴定,同时对其杀线虫特性进行表征,并通过盆栽试验进一步验证其生防潜力。结果显示,从山东省蔬菜主产区番茄根际土中分离到1株高效杀线虫菌株AMCC 100218,结合生理生化试验与16S rRNA序列分析,鉴定此菌株为和田鞘氨醇杆菌Sphingobacterium hotanense;该菌株对南方根结线虫2龄幼虫的致死效果可达88.87%,其杀线虫活性物质具有较好的热稳定性和储存稳定性,且耐碱不耐酸;盆栽试验结果表明,该菌能够显著减少土壤中的虫口密度,降低番茄发病率。表明和田鞘氨醇杆菌AMCC 100218菌株是1株具有防治根结线虫病潜力的生防细菌。     

<Emphasis Type="Italic">Solanum sisymbriifolium</Emphasis> - a new approach for the management of plant-parasitic nematodes

Plant-parasitic nematodes are serious pests causing important crop losses worldwide. After extensive screening of non-tuber-bearing Solanaceae, a resistant trap crop, Solanum sisymbriifolium, with a high production level of hatching agents, seemed an ideal control method for potato cyst nematodes (PCN), Globodera spp. Recently, root-knot nematodes (RKN), Meloidogyne spp., were found coexisting with PCN. Therefore, it is important to find alternative methods to control both nematode genera. The chemical properties of S. sisymbriifolium turns this plant into an excellent candidate for further nematicidal studies and to develop new crop production models. Studies concerning the effects of this plant on plant-parasitic nematodes are presented. Pathogenicity studies with four S. sisymbriifolium cvs (Domino, Pion, Sis 4004 and Sharp) and five Meloidogyne species showed that all cultivars of S. sisymbriifolium studied were resistant to M. chitwoodi and hypersusceptible to M. arenaria and M. hapla. For M. hispanica only cv Pion was susceptible. M. javanica induced different responses: cvs Pion and Sharp were susceptible; cv Domino resistant and Sis 4004 hypersusceptible. The studies of the hatching effects of root exudates from these cvs showed that they had an influence on the hatching inhibition of second stage juveniles of the five Meloidogyne species tested.     

我国烟草根结线虫病发生与防治研究进展

 烟草根结线虫病是一种严重影响烟叶产量和质量的重要病害之一。近年来,随着烟叶种植布局的优化调整以及大气温室效应的影响,烟草根结线虫病在全国各烟区的危害逐年加重,因此必须加强对烟草根结线虫病的发生与防治研究。本文详细综述了我国烟草根结线虫病的发生分布、危害损失、致病机制、发病规律及防治措施等研究进展,并对烟草根结线虫的鉴定技术、协同致病、抗病育种以及新型防控技术进行了总结和展望。明确了我国烟草根结线虫病优势种为南方根结线虫,并且多种根结线虫混发的现象逐步增加。同时,发现由烟草根结线虫病诱发的多种其他根茎类病害的危害逐年加重。通过加强烟草根结线虫发生动态监测、合理布局抗病品种、应用针对性的农业防治、生态防治和生物防治等综合防治措施,可以逐步实现对烟草根结线虫病的绿色防控,从而促进烟草产业的健康发展。     

A high-throughput method for early screening of coffee (<Emphasis Type="Italic">Coffea</Emphasis> spp.) genotypes for resistance to root-knot nematodes (<Emphasis Type="Italic">Meloidogyne</Emphasis> spp.)

Root-knot nematodes (Meloidogyne spp.) threaten the livelihood of millions of farmers producing coffee worldwide. The use of resistant plants either as cultivars or rootstocks appears to be the single most effective method of control. A screening method was developed to evaluate large populations of plants for resistance to root-knot nematodes. Two coffee cultivars, one susceptible and the other resistant to Meloidogyne paranaensis, were grown under controlled conditions in two substrates: a commercial sieved potting compost and an inert substrate containing sand with a water-absorbent synthetic polymer. Plant growth and development and nematode multiplication were compared for two inoculation dates (2 and 8 weeks after planting) and two evaluation dates (eight and 13 weeks after inoculation). Root growth, but not nematode multiplication, was influenced by the choice of substrate. Evaluation of the differences in root weight and nematode numbers between the different cultivars, substrates and dates of inoculation suggested that an optimal condition could be defined. The best discrimination between susceptible and resistant plants was found in the experiment where inoculation occurred at 2 weeks after planting and evaluation occurred at 8 weeks after inoculation. Because the total duration of this experiment was only 3 months, high-throughput evaluation was possible, opening up new possibilities for screening large germplasm collections and studying the genetic control of root-knot nematode resistance in coffee.     

根结线虫的研究现状

概述了根结线虫的发生分布和传统分类鉴定,以及分子生物学技术(同工酶电泳技术、DNA重组技术、PCR技术)在根结线虫种和生理小种鉴定中的运用及其防治,并对生物防治及抗根结线虫育种、转基因工程在植物线虫学研究领域上的应用前景作了展望。     

Screening new <Emphasis Type="Italic">Arachis</Emphasis> amphidiploids,and autotetraploids for resistance to late leaf spot by detached leaf technique

Cultivated groundnut is susceptible to late leaf spot (LLS) caused by Phaeoisariopsis personata [(Berk. & M.A. Curtis) Aex] and resistance is low to moderate in the primary gene pool of groundnut. Closely related wild species in the secondary gene pool are highly resistant to the disease. All the closely related Arachis species are diploid and cultivated groundnut is a tetraploid. Utilization of diploid Arachis species to transfer LLS resistance is time consuming and cumbersome. New sources of Arachis hypogaea (also called synthetic groundnut) were developed at ICRISAT. These are tetraploids and the present investigation has shown that they are resistant to LLS.     

Development of a real‐time PCR method for the detection of the dagger nematodes Xiphinema index,X. diversicaudatum,X. vuittenezi and X. italiae,and for the quantification of X. index numbers

The ectoparasitic dagger nematodes Xiphinema index and Xiphinema diversicaudatum, often at low numbers in the soil, are vectors of grapevine nepoviruses, which cause huge agronomical problems for the vineyard industry. This study reports a method, based on real‐time PCR, for the specific detection of these species and of the closely related non‐vector species Xiphinema vuittenezi and Xiphinema italiae. Specific primers and TaqMan probes were designed from the ribosomal DNA internal transcribed spacer 1 (ITS1), enabling the specific detection of single individuals of each of the X. index, X. diversicaudatum, X. italiae and X. vuittenezi species whatever the nematode population. The specificity of detection and absence of false positive reaction were confirmed in samples of each species mixed with the three other Xiphinema species or mixed with nematodes representative from other genera (non‐plant‐parasitic Dorylaimida, Longidorus sp., Meloidogyne spp., Globodera spp. and Pratylenchus sp.). The method was shown to be valid for the relative quantification of X. index numbers through its use, from crude nematode extracts of soil samples, in a greenhouse assay of grapevine accessions ranging from highly susceptible to resistant. As an alternative to time‐consuming microscopic identification and counting, this real‐time PCR method will provide a fast, sensitive and reliable diagnostic and relative quantification technique for X. index nematodes extracted from fields or controlled conditions.     

Genetic variability of Meloidogyne paranaensis populations and their aggressiveness to susceptible coffee genotypes

Meloidogyne paranaensis is one of the most destructive root‐knot nematode (RKN) species parasitizing coffee in Brazil and in the Americas generally. The objectives of this study were to assess the genetic variability, aggressiveness and virulence of seven different M. paranaensis populations on susceptible and resistant Coffea spp. All seven RKN populations were identified by biochemical and molecular methods. Coffee seedlings were inoculated in the greenhouse, and the nematode reproduction factor was used to infer their reproduction on coffee genotypes. Phylogenetic studies showed a low genetic variability in M. paranaensis populations, regardless of the existence of three esterase phenotypes (Est P1, P2 and P2a), except for the population Est P2a from Guatemala, which is genetically different from other M. paranaensis populations from Brazil. The Est P2a and Est P2 (Herculândia, SP, Brazil) populations were the most aggressive on two susceptible C. arabica cultivars under greenhouse conditions. None of the M. paranaensis populations were virulent on resistant coffee genotypes, confirming their resistance to the seven M. paranaensis populations tested. The resistant coffee cultivars, namely Clone 14 INCAPER, Catuaí Vermelho × Amphillo MR2161 (E1 16‐5 III), Apoatã IAC 2258, Timor Hybrid UFV 408‐01 (E1 6‐6 II) and IPR 100, exhibited segregation for resistance in the ratio of 0%, 2.4%, 12%, 26% and 29%, respectively. These are promising results, because they validate resistance against several M. paranaensis populations in different Coffea spp. genetic resources, which can be used in breeding programmes or as rootstocks, such as Apoatã IAC 2258 and Clone 14 INCAPER.