Novel Functions of the Fatty Acid and Retinol Binding Protein (FAR) Gene Family Revealed by Fungus-Mediated RNAi in the Parasitic Nematode, Aphelenchoides besseyi

RNA interference (RNAi) is a powerful tool for the analysis of gene function in nematodes. Fatty acid and retinol binding protein (FAR) is a protein that only exists in nematodes and plays an important role in their life activities. The rice white-tip nematode (RWTN), Aphelenchoides besseyi, is a migratory endoparasitic plant nematode that causes serious damage in agricultural production. In this study, the expression levels of eight RWTN genes were effectively decreased when RWTN was fed Ab-far-n (n: 1–8) hairpin RNA transgenic Botrytis cinerea (ARTBn). These functions of the far gene family were identified to be consistent and diverse through phenotypic changes after any gene was silenced. Such consistency indicates that the body lengths of the females were significantly shortened after silencing any of the eight Ab-far genes. The diversities were mainly manifested as follows: (1) Reproduction of nematodes was clearly inhibited after Ab-far-1 to Ab-far-4 were silenced. In addition, silencing Ab-far-2 could inhibit the pathogenicity of nematodes to Arabidopsis; (2) gonad length of female nematodes was significantly shortened after Ab-far-2 and Ab-far-4 were silenced; (3) proportion of male nematodes significantly increased in the adult population after Ab-far-1, Ab-far-3, and Ab-far-5 were silenced, whereas the proportion of adult nematodes significantly decreased in the nematode population after Ab-far-4 were silenced. (4) Fat storage of nematodes significantly decreased after Ab-far-3, Ab-far-4, and Ab-far-7 were silenced. To our knowledge, this is the first study to demonstrate that Ab-far genes affect sex formation and lipid metabolism in nematodes, which provides valuable data for further study and control of RWTNs.

The effect of ultra high-diluted drugs on plant-nematode interaction

Among plant pathogens, sedentary endoparasitic nematodes are one of the most damaging pests in global agriculture. Within this group, root-knot nematodes (RKN) Meloidogyne spp. is one of the most specialized phytoparasitic nematodes according to the complexity of the induced feeding sites. These phytopathogenic worms are highly resistant due to the large physiological variability, therefore difficult to fight against. The traditional methods of control, such as the crop rotation and the use of resistant varieties by classical selection are not always effective, time consuming and costly. The modern methods including the application of genetically modified crops (GMO), synthetic pesticides and bionematicides could be dangerous for human health and ecologically hazardous (Thomason, 1987; Malatesta et al. 2002a,b, 2008a,b; Spiroux de Vendômois et al., 2010; Seralini et al., 2012). Therefore, alternative routes must be taken in order to obtain plants resistant to nematodes. The ultra high dilutions (HDs) approach is largely in charge to dissect the infective process. HDs method is eco-friendly, low cost and leaves no residue in the environment. The classical phytopathological methods combined with modern microscopy approaches allowed to characterize the effect of different HD drugs on compatible interaction between model plant Arabidopsis thaliana and root-knot nematodes Meloidogyne incognita.

New genetic tools for plant defense against parasitic nematodes

Nematodes belong to economically important pests. Here we reviewed the recent data on molecular mechanisms of plant resistance to cyst and gall nematodes including the most devastating Globodera rostochiensis, G. pallida, Heterodera schachtii, Meloidogyne chitwoodi, and M. incognita. The Golden Potato Cyst Nematode (G. rostochiensis, GPCN) may be taken as an example of an economically important pest: in Russia, it occurs in 61 regions with a total area of 1.8 million ha and may cause the yield loss from 19 to 90 %. The biological characteristics of sedentary nematodes makes their agrotechnical control problematic, i.e. the GPCN cysts remain dormant in soil for many years until a susceptible host appears, whereas nematicides are either toxic or inefficient. Introgression of resistance genes (R-genes) from related cultivated or wild species is likely to be the most appropriate way for their biocontrol. The life cycle of sedentary nematodes is based on juveniles’ penetration into the host root where they reprogram plant cells into a syncytium or the so-called ‘giant cells’ and inhibit the plant defense response. Molecular mechanisms of plant-nematode interaction are unusual and this phenomenon provides a very interesting model for the investigation of plant morphogenesis control as well as for the development of new genetic instruments of biocontrol. Here we reviewed recent publications on plant parasitic nematode effectors used for hijacking of the plant immune system, data on R-genes and molecular mechanisms of their activities. In addition, host-induced gene silencing (HIGS) is discussed as a perspective mechanism for nematode biocontrol. HIGS is based on the RNA interference in the cells of the host plant addressed against the nematode genes important for their development and productivity. Several recent investigations demonstrated efficiency of HIGS against sedentary nematodes.

Transcriptome Analysis of Eggplant Root in Response to Root-Knot Nematode Infection

Eggplant (Solanum melongena L.), which belongs to the Solanaceae family, is an important vegetable crop. However, its production is severely threatened by root-knot nematodes (RKNs) in many countries. Solanum torvum, a wild relative of eggplant, is employed worldwide as rootstock for eggplant cultivation due to its resistance to soil-borne diseases such as RKNs. In this study, to identify the RKN defense mechanisms, the transcriptomic profiles of eggplant and Solanum torvum were compared. A total of 5360 differentially expressed genes (DEGs) were identified for the response to RKN infection. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that these DEGs are mainly involved in the processes of response to stimulus, protein phosphorylation, hormone signal transduction, and plant-pathogen interaction pathways. Many phytohormone-related genes and transcription factors (MYB, WRKY, and NAC) were differentially expressed at the four time points (ck, 7, 14, and 28 days post-infection). The abscisic acid signaling pathway might be involved in plant-nematode interactions. qRT-PCR validated the expression levels of some of the DEGs in eggplant. These findings demonstrate the nematode-induced expression profiles and provide some insights into the nematode resistance mechanism in eggplant.

Changes in the Plant β-Sitosterol/Stigmasterol Ratio Caused by the Plant Parasitic Nematode Meloidogyne incognita

Sterols play a key role in various physiological processes of plants. Commonly, stigmasterol, β-sitosterol and campesterol represent the main plant sterols, and cholesterol is often reported as a trace sterol. Changes in plant sterols, especially in β-sitosterol/stigmasterol levels, can be induced by different biotic and abiotic factors. Plant parasitic nematodes, such as the root-knot nematode Meloidogyne incognita, are devastating pathogens known to circumvent plant defense mechanisms. In this study, we investigated the changes in sterols of agricultural important crops, Brassica juncea (brown mustard), Cucumis sativus (cucumber), Glycine max (soybean), Solanum lycopersicum (tomato) and Zea mays (corn), 21 days post inoculation (dpi) with M. incognita. The main changes affected the β-sitosterol/stigmasterol ratio, with an increase of β-sitosterol and a decrease of stigmasterol in S. lycopersicum, G. max, C. sativus and Z. mays. Furthermore, cholesterol levels increased in tomato, cucumber and corn, while cholesterol levels often were below the detection limit in the respective uninfected plants. To better understand the changes in the β-sitosterol/stigmasterol ratio, gene expression analysis was conducted in tomato cv. Moneymaker for the sterol 22C-desaturase gene CYP710A11, responsible for the conversion of β-sitosterol to stigmasterol. Our results showed that the expression of CYP710A11 was in line with the sterol profile of tomato after M. incognita infection. Since sterols play a key role in plant-pathogen interactions, this finding opens novel insights in plant nematode interactions.

Screening plants for resistance/susceptibility to plant-parasitic nematodes.

This chapter provides information on the methods for initial screening to determine the resistance or susceptibility of plants, cultivars or breeding lines to plant parasitic nematodes. Specific protocols for screening and further resistance breeding on several plant-nematode combinations are described and practical guidelines for screening of Musa germplasm and several screening procedures for cyst nematodes are presented.