Professional Documents
Culture Documents
Application of Biotechnology For Nematode Control
Application of Biotechnology For Nematode Control
CITATION
READS
433
2 AUTHORS:
Michael G.K. Jones
Murdoch University
197 PUBLICATIONS 3,780 CITATIONS
SEE PROFILE
John Fosu-Nyarko
Murdoch University
24 PUBLICATIONS 235 CITATIONS
SEE PROFILE
CHAPTER FOURTEEN
Contents
1. Introduction
2. Early Selection for Plants with Nematode Resistance; Susceptibility, Resistance
and Tolerance
3. Biotechnological Approaches to Plant Parasitic Nematode Control
4. Natural Resistance Approach to Plant Parasitic Nematode Control
4.1 Transfer of Natural Resistance Genes to Different Species
5. Transgenic Approaches to Plant Parasitic Nematode Control
5.1 Disruption of Feeding Site Formation or Function
5.2 Overexpression of Host Genes with Modied Expression in Feeding Cells
5.3 RNAi-Based Nematode Resistance
5.4 Differences in Responses to RNAi in Different Nematode Species
5.5 Factors that Affect the Efcacy of RNAi Traits
5.6 Differences in Results between Model and Crop Plants
5.7 Broad Resistance to Different Plant Nematodes
6. Transgenic Technology Advances
7. From the Laboratory to the Market e Commercialization of Plant Parasitic
Nematode-Resistance Traits
7.1 Patenting
7.2 Commercialization Pathway
7.3 The Funding Gap for Early Stages of Commercialization
7.4 The Commercial Value of Nematode Resistance Traits
7.5 Specialist/Small-Scale Commercialization of Nematode Resistance Traits
8. Transgenic Nematode Resistance for Public Good
9. Regulation and Public Acceptance of GM Traits
10. Safety of RNAi-Based Traits
11. Genome-Enabled Development of Novel Chemical Nematicides
12. Ectopic Delivery of dsRNA e Nontransgenic RNAi
13. Other New Nematode Control Agents
14. Conclusions
References
359
360
362
362
363
363
365
365
366
367
367
368
371
339
340
341
344
344
348
349
349
350
350
355
356
357
357
357
359
340
Abstract
Effective control of plant parasitic nematodes in crop plants will contribute hundreds of
millions of dollars to global agriculture and help underpin future food security. Natural
nematode resistance genes present in gene pools of crop species and their relatives
have long been exploited with the aim of transferring such traits into economically
important crops where effective resistance is lacking. Biotechnology also contributes
to this process via marker-assisted selection to identify and combine the best nematode
resistance genes, and increasingly in providing new knowledge of target genes, and the
potential to exploit this knowledge using transgenic technology. Thus recent advances
now make it possible to exploit specic aspects of nematode-host plant interactions to
design control strategies that include enabling plants to prevent nematode invasion,
reducing effectiveness of nematode migration through tissues, preventing successful
establishment or reducing feeding ability or nematode fecundity. The knowledge of
what genes are vital for successful nematode parasitism can also be used to develop
new chemical control agents. These new strategies may either be available for public
use or be delivered commercially. For transgenic technologies, both modes of delivery
face the same issues in terms of deployment, such as substantial eld testing, meeting
environmental and human safety regulations, adequate funding to complete statutory
requirements, and public acceptance of GMOs when the product is to be marketed.
However, as technology develops, new strategies for nematode control are emerging,
both for transgenic approaches and in genome editing, which should be regarded by
regulators as a form of mutation rather than genetic modication. With such advances
in biotechnology, the release of commercial varieties of major crops with new forms of
nematode resistance, or new modes of delivery of control agents, is likely to become a
commercial reality. To improve durability, transgenic traits could be based on resistance
with different modes of action: for example, RNAi-based technology combined with
expression of peptides which disrupt sensory activities. Ideally such traits would be
added to existing crop genotypes with the best conventional or natural nematode resistance, to increase the effectiveness and durability of the nematode resistance trait.
Biotech trait expression could also be limited to roots to minimise expression in harvested parts, and this could improve public acceptability.
1. INTRODUCTION
The current status of molecular understanding of nematodeplant interactions is described in earlier chapters in this volume, and it is clear that
rapid advances are being made in unravelling the mechanisms which enable
plant parasitic nematodes to be such successful plant pests. The question
addressed in this chapter is how this new information can be translated to practical application, and used to reduce crop losses caused by these devastating
parasites. If this can be achieved it will be a signicant contribution to future
341
crop security and increased productivity in a sustainable manner. The requirement to nd new ways of controlling plant nematodes is all the more pressing
because some of the older chemical nematicides have been withdrawn or are
now under restricted use mandates: this has led to a renewed interest in developing new strategies to control plant parasitic nematodes based on genetic,
chemical or integrated approaches to manage nematode pests.
The academic advances in knowledge are now impressive, and it is clear
that research of excellent quality is being done to understand nematodeplant
interactions: in particular such research is leading to identifying what effectors
they secrete to be able to avoid or neutralize host plant defences, detect gradients, migrate within roots and, depending on the species, induce the formation
of long-term feeding sites. However, there is still a gap between this basic
research and its practical application to control these pests. As concluded by
McCarter (2009), the future of plant nematology as a discipline is dependent
on the value of commercial solutions delivered to growers. Such advances
are likely to come from both conventional and genetic approaches: McCarter
also emphasized that economically and environmentally sound methods to
control nematodes which contribute a commercial increase in crop yields
will result in more investment in the eld. A summary of the biotechnology-based strategies now available for nematode control, which include
both established breeding technologies and transgenic approaches, is provided
in Table 1, with brief explanations of the strategies and of their current status.
Status/Example
342
John Fosu-Nyarko and Michael G.K. Jones
343
344
345
Heterodera schachtii
Heterodera avenae
Globodera pallida
Heterodera glycines
Globodera rostochiensis
Cyst Nematodes
Potato
Potato
Tomato
Potato
Soybean
Soybean
Barley
Barley
Wheat
Wheat
Wheat
Sugar beet
Sugar beet
Gro1
H1
Hero
Gpa2
rhg1
Rhg4
Ha2, Ha3
Ha4
Cre1
Cre3
Cre8
Hs1pro1
Hs2
Table 2 Summary of Natural Resistance Genes to Cyst and Root Knot Nematodes, and Major QTLs Associated with Resistance to
Pratylenchus spp
Crop or Source
Nematode Species
Resistance Genes
of Resistance
References
346
John Fosu-Nyarko and Michael G.K. Jones
Mi-1
Mi-3 on chromosome 12
Mi-9
Meloidogyne incognita
Pratylenchus thornei
Pratylenchus neglectus
Peanut
Me3 on chromosome P9
Meloidogyne arenaria
347
348
Macrosiphum euphorbiae and the white y Bemisia tabaci (Nombela, Williamson, & Mu~
niz, 2003; Rossi et al., 1998). Although Pratylenchus species are
often regarded as being less damaging in terms of crop losses, they can be
the most economically important nematode pests in areas of low rainfall
such as the Australian wheatbelt. Not surprisingly, the most detailed
research on breeding for tolerance and resistance to Pratylenchus spp. has
been carried out in Australian cereal breeding programs (Table 2) (Jones
& Fosu-Nyarko, 2014). Genotypes with high tolerance to infestation
with Pratylenchus thornei and medium tolerance to Pratylenchus penetrans
have been identied among wheat cultivars, although they are not necessarily resistant or tolerant to other Pratylenchus species (Smiley & Nicol,
2009). Also major Quantitive Trait Loci (QTLs) for P. thornei, P. penetrans
and Pratylenchus neglectus, some of which have polygenic and additive resistance effects, have been used routinely to select for resistance for these
nematodes in Australian and CIMMYT (International Maize and Wheat
Improvement Center) wheat breeding programs (Table 2) (Williams
et al., 2002).
Study of resistance to Pratylenchus species is also important in barley
because losses caused can also be substantial. To date, ve QTL loci contributing to resistance to P. neglectus in barley germplasm have been identied on
chromosomes 3H, 5H, 6H and 7H, and these may be useful for markerassisted selection for resistance in barley (Table 2) (Sharma et al., 2011).
Although the resistance conferred by some of these genes is useful in
improving resistance to Pratylenchus species in commercial crop varieties,
there is still a need to identify new, more effective and durable sources of
natural resistance to nematodes in most major crop species.
349
350
351
to current methods used for nematode control (Fire et al., 1998; Tan, Jones, &
Fosu-Nyarko, 2013; Urwin, Lilley, & Atkinson, 2002). The rst question
was how to make plant parasitic nematodes take up dsRNA from external
solutions, and this was solved by the pioneering work of Urwin et al.
(2002), who showed that upon addition of neurostimulants to the soaking
solution for H. glycines J2s they could be induced to take up sufcient dsRNA
to induce RNAi. Since that time, dsRNA feeding/soaking has been used to
assess the effects of downregulation of over 30 essential and parasitism genes
of various plant nematode species, including cyst nematodes (H. glycines, H.
schachtii, Gobodera pallida, Gobodera rostochiensis), root knot nematodes (M.
incognita, M. javanica, Meloidogyne hapla, M. arenaria and Meloidogyne artiellia),
root lesion nematodes (Pratylenchus zeae, P. thornei, Pratylenchus coffeae) and
other ectoparasitic nematodes (Radopholus similis, and Bursaphelenchus xylophilus) (Joseph, Gheysen, & Subramaniam, 2012; Li, Todd, Oakley, Lee, &
Trick, 2011; Lilley, Bakhetia, Charlton, & Urwin, 2007; Tan et al., 2013)
(Reviewed for RKNs in chapter Function of Root-Knot Nematode Effectors and Their Targets in Plant Parasitism). However, it has since been
demonstrated that neurostimulants and other chemicals are not necessarily
needed to induce RNAi using dsRNA (e.g. Fanelli, Di, Jones, & Giorgi,
2005; Kimber et al., 2007). In H. glycines and for some Pratylenchus spp, it appears that, for some genes at least, silencing resulting from soaking in dsRNA
does not always produce stable phenotypic effects, since it appears that the
effects of RNAi can wear off hours or days after the initial effect, leading
to nematode recovery or regaining of function. Nevertheless the soaking
method has been shown to be an effective method for initial screening of
gene function and for discovery of candidate target genes suitable for
plant-delivered RNAi for nematode control.
In contrast to soaking plant nematodes in solutions containing dsRNA,
host (in planta) delivery provides dsRNA continuously if expressed in host
cells from a constitutive promoter. This mode of delivery of dsRNA appears
to be an ideal and economical approach to control obligate parasites such as
plant parasitic nematodes. In planta delivery of dsRNA of two target genes
(an integrase and a pre-mRNA splicing factor) was rst demonstrated by
Yadav, Veluthambi, and Subramaniam (2006) to reduce replication of M.
incognita on transgenic tobacco plants, and this was quickly followed by
the work of Huang, Allen, Davis, Baum, and Hussey (2006) who expressed
dsRNA to an M. incognita effector protein in transgenic plants and also
showed reduced nematode reproduction. Since then a series of experimental and crop plants have been engineered to generate inverted repeats
Meloidogyne
incognita
Tomato
Grapes
Tomato
Soybean
Troponin C (tnc)
Secreted peptide (16D10)
Calreticulin (crt)
L-Lactate
Soybean
Arabidopsis
dehydrogenase
Tobacco
Tobacco
Table 3 Host-Delivered RNAi for Parasitism and Essential Genes of Cyst and Root Knot Nematodes: Summary of Reduced Infectivity on
Model and Crop Plants
Nematode
Gene Silenced
Plant/Crop
Major Phenotype
References
352
John Fosu-Nyarko and Michael G.K. Jones
Arabidopsis
Potato
Meloidogyne
hapla
Meloidogyne
chitwoodi
Arabidopsis
Arabidopsis
Arabidopsis
Meloidogyne
arenaria
Tomato
Tomato
Dual oxidase
Soybean
Tyrosine phosphatase
Meloidogyne
javanica
Soybean
(Continued)
353
Heterodera
schachtii
Heterodera
glycines
Up to 68% reduction in female cysts
87% reduction in female cysts
81% reduction in female cysts
88% reduction in female cysts
93% reduction in female cysts
81% reduction in female cysts
79% reduction in nematode
95% reduction in nematode
23e64% reduction in developing
females
12e47% reduction in developing
females
>50% reduction in developing females
42% reduction in developing females
36% reduction in female cysts
Up to 20% reduction in female cysts
Up to 20% reduction in female cysts
Up to 55% reduction in female cysts
Up to 92% reduction in female cysts
Soybean
Soybean
Soybean
Soybean
Soybean
Soybean
Soybean
Soybean
Arabidopsis
Arabidopsis
Arabidopsis
Arabidopsis
Arabidopsis
Arabidopsis
Arabidopsis
Arabidopsis
Arabidopsis
Cyst Nematodes
Li et al. (2010)
Li et al. (2010)
Sindhu et al. (2009)
Table 3 Host-Delivered RNAi for Parasitism and Essential Genes of Cyst and Root Knot Nematodes: Summary of Reduced Infectivity on
Model and Crop Plantsdcont'd
Nematode
Gene Silenced
Plant/Crop
Major Phenotype
References
354
John Fosu-Nyarko and Michael G.K. Jones
355
356
et al., 2008; Sindhu et al., 2009; Patel et al., 2010; Hamamouch et al., 2012).
However, silencing seven genes (elongation factor 1a, two vacuolar H
ATPases, integrase, pre-mRNA splicing factor, troponin C and tropomyosin) of H. schachtii via transgenic Arabidopsis resulted in up to 98% reduction
in adult females (Fosu-Nyarko & Jones, 2013, 2014).
The differences in effectiveness of RNAi may relate to differences in
biology of hostparasite interactions such as presence or absence of feeding
tubes: Meloidogyne feeding tubes appear to be larger and more regular in
structure than those formed by feeding cyst nematodes, and this may inuence the ability of uptake of dsRNAs, whereas Pratylenchus species do not
form feeding tubes. Alternatively there may be fundamental differences in
RNAi pathways in different nematodes, or in systemic movement of siRNAs in the nematodes. The reasons for such differences, or whether current
reports reect more differences in experimental procedures used, remain to
be demonstrated experimentally.
357
plant events will exhibit a range of the desired property, and only the most
effective events will be chosen for progression through a commercialization
pipeline.
358
359
360
one or more traits. In the latter case, company investors will seek a way to
recoup their investment (an exit strategy), either by trade sale to a larger
company, by public listing or by raising further investment to exploit the
trait directly. The further the product is progressed along the developmental
pathway, the higher the expected returns will be.
361
Discovery
Early discovery
Late discovery
Total cost
Construct optimization
Commercial Event production & selection
Introgression Breeding & Wide Area Testing
Regulatory Science
Regulatory & Regulatory Affairs
Total
17.6
13.4
31.0
28.3
13.6
28.0
17.9
17.2
136.0
5
5
5
5
6
6
6
6
362
363
364
365
366
367
368
molecules that will t the active site of established target gene products and
inhibit their vital function.
One focus is on an integrated pest management strategy to control nematodes, which may combine both biological and chemical agents, agents such
as Bayer CropSciencess Bacillus rmus (Votivo), Paecilomyces lilacinus (Bioact)
combined with chemical control, e.g. uopyram (Velum/Verango), or using Pasteuria technology (e.g. CALARIVA) with AVICTA (Syngenta) and
other new products on the market. These can be combined with more
restricted methods of delivery, such as via drip irrigation or seed coating.
These delivery methods are more suited to nematode control in horticultural crops, and have the benet of providing control agents only where they
are needed, thus enabling more expensive and less stable chemicals to be
applied, so reducing costs and any detrimental environmental effects.
14. CONCLUSIONS
The exploitation of natural resistance genes using genotype screening
and marker-assisted selection will continue to be the standard approach to
improving resistance and/or tolerance to nematodes for many horticultural
369
and staple food crops. However, for many crops this approach is slow, and in
most cases adequate broad-spectrum resistance is not available. This in part
accounts for the lag in approved release of nematode resistance traits
compared to other important agronomic traits as compiled by Information
Systems for Biotechnology (Figure 3, http://www.isb.vt.edu/releasesummary-data.aspx). With advances in transgenic technologies the release
of commercial varieties of major crops with transgenic nematode resistance
traits can be predicted to arrive soon. Already maize with eight stacked genes
(SmartStax Geniutycorn, with two herbicide tolerances and six Bacillus
thuringiensis-based insect resistance traits) is commercially available, and varieties with similar stacked traits will soon be available for cotton, soybean and
other crops. The addition of a nematode resistance trait would differentiate
the rst producer of such a trait, and would provide a competitive advantage. Such a biotech trait could be based on RNAi or on expression of other
synthetic resistance genes, and be added to genotypes with the best conventional nematode resistance. In addition, synthetic resistance genes with
different modes of action could be stacked to increase the efcacy and durability of the nematode resistance trait, and their expression limited to roots to
improve acceptability and unnecessary expression in harvested parts of the
plant.
As emphasized by Atkinson et al. (2012), crop losses to nematode infestation is often greatest in developing countries, where the need for food
security is greatest, and nematodes attack both commodity and staple crops.
In these countries it may not be a commercial proposition to develop nematode-resistant transgenic crops: rather a humanitarian approach may be
adopted in which such crops are developed for the public good. However,
following this approach will still require detailed studies to be undertaken to
ensure the food is safe for humans and the environment, and the political and
public support for their implementation, and in country support for their
further development and deployment.
Since two-thirds of the worlds population already lives in countries
where GM crops are grown, and concern for world food security is
now increasing, it is likely that the pace for adoption of transgenic
crops will increase as one of the mix of measures that will be needed
to feed over 9 billion people in a sustainable manner. Transgenic nematode resistance traits will undoubtedly be used to help ensure global food
security in the future, although the path to reach this stage will have
some issues to address, particularly in relation to public perception rather
than science.
370
That is not to say that there have not been practical successes in
nematode control, as can be seen when considering advances in control
for different crops and nematode pests on a case-by-case basis. For
example, real benets have been derived through conventional plant
breeding practices by introduction of the H1 major resistance gene
against the potato cyst nematode Globodera rostochiensis (FinkersTomczak et al., 2011) and the Mi gene conferring host resistance to
root knot (Meloidogyne) species in tomato (Williamson & Kumar, 2006),
which has been transferred to many different tomato varieties throughout
the world. A notable success has arisen from the study of natural genes for
resistance to the cereal cyst nematode (H. avenae), in which effective control has been developed for wheat initially using large-scale screening of
genotypes with resistance to H. avenae, followed by identication of
NBS-LRR type resistance genes from wheat and its wild relatives, and
their implementation in breeding programs. A series of 11 Cre genes
have been identied and introgressed into wheat cultivars (in particular
the genes Cre 1, Cre3 and Cre 8) to confer resistance, particularly in
Australia (Lewis, Matic, & McKay, 2009; Ogbonnaya, Eastwood, &
Lagudah, 2009). Marker-assisted selection is now used to pyramid some
of these resistance genes, and this approach is used routinely in many
wheat breeding programs worldwide (Ogbonnaya et al., 2009) to provide
effective control of H. avenae.
Marker-assisted selection can also be used to pyramid and combine
different desired traits into improved germplasm, and this has been an
ongoing activity undertaken by plant breeders over many years. In
addition to major gene resistances, minor (multigene-based) resistance
has been used incrementally to improve host resistance to nematode
infestation in many crops, such as the rht genes in soybean improving
resistance to the soybean cyst nematode (H. glycines), and a range of
minor genes contributing to increased resistance to root lesion nematodes
(Pratylenchus spp.) in cereals (Jones & Fosu-Nyarko, 2014). The more
conventional application of biotechnology to nematode control by
marker-assisted selection is now being complemented by the developing
biotechnological approaches of transgenic plants, genome editing, developing new nematode control agents and new modes of delivery of control
agents. The commercial implementation and delivery of these biotechnology-based technologies will undoubtedly contribute to future global
food security.
371
REFERENCES
Alkharouf, N. W., Klink, V. P., Chouikha, I. B., Beard, H. S., MacDonald, M. H.,
Meyer, S., et al. (2006). Time course microarray analyses reveals global changes in
gene expression of susceptible Glycine max (soybean) roots during infection by Heterodera
glycines (soybean cyst nematode). Planta, 224, 838852.
Ammiraju, J., Veremis, J., Huang, X., Roberts, P., & Kaloshian, I. (2003). The heat-stable
root-knot nematode resistance gene Mi-9 from Lycopersicon peruvianum is localized on
the short arm of chromosome 6. Theoretical and Applied Genetics, 106, 478484.
Atkinson, H. J., Lilley, C. J., & Urwin, P. E. (2012). Strategies for transgenic nematode control
in developed and developing world crops. Current Opinion in Biotechnology, 23, 251256.
Atkinson, H. (2014). Transgenic nematode resistance for African food security: nematode
resistant bananas as a case study. Proceeding of the 6th International Congress of Nematology, Cape Town, May 2014.
Ballvora, A., Hesselbach, J., Niewoehner, J., Leister, D., Salamini, F., & Gebhardt, C. (1995).
Marker enrichment and high resolution map of the segment of potato chromosome VII
harbouring the nematode resistance gene Gro1. Molecular & General Genetics, 249, 8290.
Barcala, M., Garcia, A., Cabrera, J., Casson, S., Lindsey, K., Favery, B., Garcia-Casado, G.,
Solano, R., Fenoll, C., Escobar, C., et al. (2010). Early transcriptomic events in microdissected Arabidopsis nematode-induced giant cells. Plant J, 61, 698712.
Barr, A. R., Chalmers, K. J., Karakousis, A., Kretschmer, J. M., Manning, S.,
Lance, R. C. M., et al. (1998). RFLP mapping of a new cereal cyst nematode resistance
locus in barley. Plant Breeding, 117, 185187.
Breyer, D., Herman, P., Brandenburger, A., Gheysen, G., Remaut, E., Soumillion, P., et al.
(2009). Genetic modication through oligonucleotide-mediated mutagenesis. A GMO
regulatory challenge? Environmental Biosafety Research, 18.
Cai, D., Kleine, M., Kie, S., Harloff, H. J., Sandal, N. N., Marcker, K. A., et al. (1997).
Positional cloning of a gene for nematode resistance in sugar beet. Science, 275, 832834.
Charlton, W. C., Harel, H. Y. M., Bakhetia, M., Hibbard, J. K., Atkinson, H. J., &
McPherson, M. J. (2010). Additive effects of plant expressed double-stranded RNAs
on root-knot nematode development. International Journal for Parasitology, 40, 855864.
Chitwood, D. J. (2002). Phytochemical based strategies for nematode control. Annual Review
of Phytopathology, 40, 221249.
Chu, Y., Wu, C. L., Holbrook, C. C., Tillman, B. L., Person, G., & Ozias-Akins, P. (2011).
Marker-assisted selection to pyramid nematode resistance and the high oleic trait in
peanut. The Plant Genome, 4, 110117.
Concibido, V. C., Diers, B. W., & Arelli, P. R. (2004). A decade of QTL mapping for cyst
nematode resistance in soybean. Crop Science, 44, 11211131.
Danchin, E. G. J., Arguel, M.-J., Campan-Fouriner, A., Perfus-Barbeoch, L., Magliano, M.,
Rosso, M. N., et al. (2013). Identication of novel target genes for safer and more specic
control of root-knot nematodes from a pan-genome mining. PLoS Pathogens, 9(10),
e1003745.
Dinh, P. T. Y., Brown, C. R., & Elling, A. A. (2014). RNA interference of effector gene
Mc16D10L confers resistance against Meloidogyne chitwoodi in Arabidopsis and potato.
Phytopathology, 104, 10981106.
Djian-Caporalino, C., Fazari, A., Arguel, M. J., Vernie, T., VandeCasteele, C., Faure, I., et al.
(2007). Root-knot nematode (Meloidogyne spp.) Me resistance genes in pepper (Capsicum annuum L.) are clustered on the P9 chromosome. Theoretical and Applied Genetics, 114, 473486.
Dubreuil, G., Magliano, M., Dubrana, M. P., Lozano, K., Lecomte, P., Favery, B., Abad, P.,
& Rosso, M. N. (2009). Tobacco rattle virus mediates gene silencing in a plant parasitic
root-knot nematode. Journal of Experimental Botany, 60, 40414050.
372
Eastwood, R. F., Lagudah, E. S., & Appels, R. (1994). A directed search for DNA sequences
tightly linked to cereal cyst nematode resistance genes in Triticum tauschii. Genome, 37,
311319.
Escobar, C., Barcala, M., Portillo, M., Almoguera, C., Jordano, J., & Fenoll, C. (2003). Induction of the Hahsp17.7G4 promoter by root-knot nematodes: involvement of heatshock elements in promoter activity in giant cells. Molecular Plant-Microbe Interaction,
16, 10621068.
Fanelli, E., Di, V. M., Jones, J. T., & Giorgi, C. (2005). Analysis of chitin synthase function in
a plant parasitic nematode, Meloidogyne artiellia, using RNAi. Gene, 349, 8795.
FAOSTAT. (2012). FAO Statistical Year Book, 2012. Rome: Food and Agriculture Organization of the United Nations, ISBN 978-92-5-106913-4.
Finkers-Tomczak, A., Bakker, E., Boer, J. D., Van der Vossen, E. A., Achenbach, U.,
Golas, T., et al. (2011). Comparative sequence analysis of the potato cyst nematode resistance locus H1 reveals a major lack of co-linearity between three haplotypes in potato
(Solanum tuberosum ssp). Theoretical Applied Genetics, 122, 595608.
Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver, S. E., & Mello, C. C. (1998).
Potent and specic genetic interference by double-stranded RNA in Caenorhabditis
elegans. Nature, 391, 806811.
Fosu-Nyarko, J., & Jones, M. G. K. (2013). Target genes for control of plant parasitic nematodes and
use of same. Patent Publication Number US20130269057.A1, PCT number PCT/
IB2011/002130.
Fosu-Nyarko, J., & Jones, M. G. K. (2014). Target genes for control of plant parasitic nematodes and
use of same. Patent Publication Number WO2014001893.A3, PCT number PCT/
IB2013/001385.
Fosu-Nyarko, J., Jones, M. G. K., & Wang, Z. (2009). Functional characterisation of transcripts in early stage Meloidogyne javanica-induced giant cells isolated by laser
microdissection. Molecular Plant Pathology, 10, 237248.
Galal, A., Sharma, S., Abou-Elwafa, S. F., Sharma, S., Kopisch-Obuch, F., Laubach, E., et al.
(2014). Comparative QTL analysis of root lesion nematode resistance in barley. Theoretical and Applied Genetics, 127, 13991407.
Ganal, M. W., Simon, R., Brommonschenkel, S., Arndt, M., Phillips, M. S., Tanksley, S. D.,
et al. (1995). Genetic mapping of a wide spectrum nematode resistance gene (Hero)
against Globodera rostochiensis in tomato. Molecular Plant Microbe Interactions, 8, 886891.
Ganal, M. W., & Tanksley, S. D. (1996). Recombination around the Tm2a and Mi resistance
genes in different crosses of Lycopersicon peruvianum. Theoretical and Applied Genetics, 92,
101108.
Garcia, G. M., Stalker, H. T., Shroeder, E., & Kochert, G. (1996). Identication of RAPD,
SCAR, and RFLP markers tightly linked to nematode resistance genes introgressed from
Arachis cardenasii into Arachis hypogaea. Genome, 39, 836845.
Hamamouch, N., Li, C., Hewezi, T., Baum, T. J., Mitchum, M. G., Hussey, R. S.,
Vodkin, L. O., & Davis, E. L. (2012). The interaction of the novel 30C02 cyst nematode
effector protein with a plant beta-1,3 endoglucanase may suppress host defence to promote parasitism. Journal of Experimental Botany, 63, 36833695.
Heller, R., Schondelmaier, J., Steinr
ucken, G., & Jung, C. (1996). Genetic localization of
four genes for nematode (Heterodera schachtii Schm.) resistance in sugar beet (Beta vulgaris
L.). Theoretical and Applied Genetics, 92, 991997.
Huang, G., Allen, R., Davis, E. L., Baum, J. T., & Hussey, S. R. (2006). Engineering broad
root-knot resistance in transgenic plants by RNAi silencing of a conserved and essential
root-knot nematode parasitism gene. Proceedings of National Academy of Science of the USA,
103, 1430214306.
Ibrahim, H. M. M., Hosseini, P., Alkharouf, N. W., Hussein, E. H. A., El Kader, A., ElDin, G., et al. (2011). Analysis of gene expression in soybean (Glycine max) roots in
373
response to the root knot nematode Meloidogyne incognita using microarrays and KEGG
pathways. BMC Genomics, 12, 220.
Jacobsen, E., & Schouten, H. J. (2008). Cisgenesis, a new tool for traditional plant breeding,
should be exempted from the regulation on genetically modied organisms in a step by
step approach. Potato Research, 51, 7588.
Jacquet, M., Bongiovanni, M., Martinez, M., Verschave, P., Wajnberg, E., & CastagnoneSereno, P. (2005). Variation in resistance to the root-knot nematode Meloidogyne incognita
in tomato genotypes bearing the Mi gene. Plant Pathology, 54, 9399.
Jones, M. G. K. (1981). Host cell responses to endoparasitic nematodes. Annals of Applied
Biology, 97, 353372.
Jones, M. G. K., & Fosu-Nyarko, J. (2014). Molecular biology of root lesion nematodes (Pratylenchus spp.) and their interaction with host plants. Annals of Applied Biology, 164, 163181.
Joseph, S., Gheysen, G., & Subramaniam, K. (2012). RNA interference in Pratylenchus coffeae:
knock down of Pc-pat-10 and Pc-unc-87 impedes migration. Molecular and Biochemical
Parasitology, 186, 5159.
Kimber, M. J., McKinney, S., McMaster, S., Day, T. A., Fleming, C. C., & Maule, A. G.
(2007). p Gene distribution in a parasitic nematode reveals motor dysfunction and unusual neuronal sensitivity to RNA interference. The FASEB Journal, 21, 12331243.
Klein-Lankhorst, R., Rietveld, P., Machiels, B., Verkerk, R., Weide, R., Gebhardt, C., et al.
(1991). RFLP markers linked to the root knot nematode resistance gene Mi in tomato.
Theoretical and Applied Genetics, 81, 661667.
Klink, V. P., Kim, K. H., Martins, V., Macdonald, M. H., Beard, H. S., Alkharouf, N. W.,
et al. (2009). A correlation between host-mediated expression of parasite genes as tandem
inverted repeats and abrogation of development of female Heterodera glycines cyst formation during infection of Glycine max. Planta, 230, 5371.
Kreike, C. M., deKoning, J. R. A., Vinke, J. H., van Ooijen, J. W., Gebhardt, C., &
Stiekema, W. J. (1993). Mapping of loci involved in quantitatively inherited resistance
to the potato cyst-nematode Globodera rostochiensis pathotype Ro1. Theoretical Applied Genetics, 87, 464470.
Kretschmer, J. M., Chalmers, K. J., Manning, S., Karakousis, A., Barr, A. R.,
Islam, A. K. M. R., et al. (1997). RFLP mapping of the Ha 2 cereal cyst nematode resistance gene in barley. Theoretical and Applied Genetics, 94, 10601064.
Lagudah, E. S., Moullet, O., & Appels, R. (1997). Map-based cloning of a gene sequence
encoding a nucleotide-binding domain and a leucine-rich region at the Cre3 nematode
resistance locus of wheat. Genome, 40, 659665.
Leister, D., Ballvora, A., Salamini, F., & Gebhardt, C. (1996). A PCRdbased approach for
isolating pathogen resistance genes from potato with potential for wide application in
plants. Nature Genetics, 14, 421429.
Lewis, J. G., Matic, M., & McKay, A. C. (2009). Success of cereal cyst nematode resistance
in Australia: history and status of resistance screening systems. In I. T. Riley, J. M. Nicol,
& A. A. Dababat (Eds.), Proceedings of the rst workshop of the international cereal cyst nematode initiative: Cereal cyst nematode: Status, research and outlook (pp. 137142) (Antalya,
Turkey).
Lilley, C. J., Bakhetia, M., Charlton, W. L., & Urwin, P. E. (2007). Recent progress in the
development of RNA interference for plant parasitic nematodes. Molecular Plant Pathology, 2007, 701711.
Li, J., Todd, T. C., Oakley, T. R., Lee, J., & Trick, H. N. (2010). Host-derived suppression
of nematode reproductive and tness genes decreases fecundity of Heterodera glycines
Ichinohe. Planta, 232, 775785.
Li, J., Todd, T. C., Oakley, T. R., Lee, J., & Trick, H. N. (2011). Biotechnological application of functional genomics towards plant-parasitic nematode control. Plant Biotechnology
Journal, 9, 936944.
374
Lin, B., Zhuo, K., Wu, P., Cui, R., Zhang, L. H., & Liao, J. (2013). A novel effector protein,
MJ-NULG1a, targeted to giant cell nuclei plays a role in Meloidogyne javanica parasitism.
Molecular Plant-Microbe Interactions, 26, 5566.
Liu, S., Kandoth, P. K., Warren, S. D., Yeckel, G., Heinz, R., Alden, J., et al. (2012). A soybean cyst nematode resistance gene points to a new mechanism of plant resistance to
pathogens. Nature, 492, 256260.
Lozano-Juste, J., & Cutler, S. R. (2014). Plant genome engineering in full bloom. Trends in
Plant Science, 19, 284287.
Matthews, B. F., Beard, H., MacDonald, M. H., Kabir, S., Youssef, R. M., Hosseini, P., et al.
(2012). Engineered resistance and hypersusceptibility through functional metabolic
studies of 100 genes in soybean to its major pathogen, the soybean cyst nematode. Planta,
237, 13371357.
McCarter, J. P. (2009). Molecular approaches toward resistance to plant-parasitic nematodes. In
R. H. Berg, & C. G. Taylor (Eds.), Cell biology of plant nematode Paraistism (pp. 239268).
Springer-Verlag Berlin, Heidelberg.
Messeguer, R., Ganal, M., De Vicente, M. C., Young, N. D., Bolkan, H., & Tanksley, S. D.
(1991). High resolution RFLP map around the root knot nematode resistance gene (Mi)
in tomato. Theoretical and Applied Genetics, 82, 529536.
Nekrasov, V., Staskawicz, B., Weigel, D., Jones, J. D. G., & Kamoun, S. (2013). Targeted
mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided
endonuclease. Nature Biotechnology, 31, 691693.
Niew
ohner, J., Salamini, F., & Gebhardt, C. (1995). Development of PCR assays diagnostic
for RFLP marker alleles closely linked to alleles Gro1 and H1, conferring resistance to the
root cyst nematode Globodera rostochiensis in potato. Molecular Breeding, 1, 6567.
Nombela, G., Williamson, V. M., & Mu~
niz, M. (2003). The root-knot nematode resistance
gene Mi-1.2 of tomato is responsible for resistance against the whitey Bemisia tabaci.
Molecular Plant-Microbe Interactions, 16, 645649.
Ogbonnaya, F. C., Eastwood, R. F., & Lagudah, E. (2009). Identication and utilisation of
genes for cereal cyst nematode resistance (Heterodera avenae) resistance in wheat: the
Australian experience. In I. T. Riley, J. M. Nicol, & A. A. Dababat (Eds.), Proceedings
of the rst workshop of the international cereal cyst nematode initiative: Cereal cyst nematode:
Status, research and outlook (pp. 166171) (Antalya, Turkey).
Opperman, C. H., Taylor, C. G., & Conkling, M. A. (1994). Root-knot nematode-directed
expression of a plant root-specic gene. Science, 263, 221223.
Patel, N., Hamamouch, N., Li, C., Hussey, R., Mitchum, M., Baum, T., Wang, X., &
Davis, E. L. (2008). Similarity and functional analyses of expressed parasitism genes in
Heterodera schachtii and Heterodera glycines. Journal of Nematology, 40, 299310.
Patel, N., Hamamouch, N., Li, C., Hewezi, T., Hussey, R., Baum, T., Mitchum, M., &
Davis, E. L. (2010). A nematode parasitism protein similar to annexins in host plants. Journal of Experimental Botany, 61, 235248.
Petrick, J. S., Brower-Toland, B., Jackson, A. L., & Kier, L. D. (2013). Safety assessment of
food and feed from biotechnology-derived crops employing RNA-mediated gene regulation to achieve desired traits: a scientic review. Regulatory Toxicology and Pharmacology,
66, 167176.
Portillo, M., Cabrera, J., Lindsey, K., Topping, J., Andrs, M. F., Emiliozzi, M., Oliveros, J. C.,
Garca-Casado, G., Solano, R., Koltai, H., Resnick, N., Fenoll, C., & Escobar, C. (2013).
Distinct and conserved transcriptomic changes during nematode-induced giant cell development in tomato compared with Arabidopsis: a functional role for gene repression. New
Phytol., 197, 12761290. http://dx.doi.org/10.1111/nph.12121.
Ramsay, K., Wang, Z. H., & Jones, M. G. K. (2004). Using laser capture microdissection to
study gene expression in early stages of giant cells induced by root-knot nematodes. Molecular Plant Pathology, 5, 587592.
375
Rossi, M., Goggin, F. L., Milligan, S. B., Kaloshian, I., Ullman, D. E., &
Williamson, V. M. (1998). The nematode resistance gene Mi of tomato confers resistance against the potato aphid. Proceedings of the National Academy of Sciences of the USA,
95, 97509754.
Schmidt, A. L., McIntyre, C. L., Thompson, J., Seymour, N. P., & Liu, C. J. (2005). Quantitative trait loci for root lesion nematode (Pratylenchus thornei) resistance in middleeastern landraces and their potential for introgression into Australian bread wheat.
Crop and Pasture Science, 56, 10591068.
Sharma, S., Sharma, S., Kopisch-Obuch, F. J., Keil, T., Laubach, E., Stein, N., et al. (2011).
QTL analysis of root-lesion nematode resistance in barley: 1. Pratylenchus neglectus. Theoretical and Applied Genetics, 122, 13211330.
Sijmons, P. C., Atkinson, H. J., & Wyss, U. (1994). Parasitic strategies of root-knot
nematodes and associated host cell responses. Annual Review of Phytopathology, 32,
235259.
Sindhu, A. S., Maier, T. R., Mitchum, M. G., Hussey, R. S., Davis, E. L., & Baum, T. J.
(2009). Effective and specic in planta RNAi in cyst nematodes: expression interference of four parasitism genes reduces parasitic success. Journal of Experimental Botany,
60, 315324.
Smiley, R. W., & Nicol, J. M. (2009). Nematodes which challenge global wheat production.
In B. F. Carver (Ed.), Wheat science and trade (pp. 171187). USA: Wiley-Blackwell.
Sobczak, M., Avrova, A., Jupowicz, J., Phillips, M. S., Ernst, K., & Kumar, A. (2005). Characterization of susceptibility and resistance responses to potato cyst nematode (Globodera
spp.) infection of tomato lines in the absence and presence of the broad-spectrum nematode resistance Hero gene. Molecular Plant-Microbe Interactions, 18, 158168.
Steeves, R. M., Todd, T. C., Essig, J. S., & Trick, H. N. (2006). Transgenic soybeans expressing siRNAs specic to a major sperm protein gene suppress Heterodera glycines
reproduction. Functional Plant Biology, 33, 991999.
Szakasits, D., Heinen, P., Wieczorek, K., Hofmann, J., Wagner, F., Kreil, D. P., et al. (2009).
The transcriptome of syncytia induced by the cyst nematode Heterodera schachtii in
Arabidopsis roots. The Plant Journal, 57(5), 771784. http://dx.doi.org/10.1111/j.
1365-313X.2008.03727.x. [pii]: TPJ3727.
Tan, J., Jones, M. G. K., & Fosu-Nyarko, J. (2013). Gene silencing in root lesion nematodes
(Pratylenchus spp) signicantly reduces reproduction in a plant host. Experimental Parasitology, 133, 166178.
Taylor, J. R. N., Belton, P. S., Beta, T., & Duodu, K. G. (2014). Increasing the utilisation of
sorghum, millets and pseudocereals: developments in the science of their phenolic
phytochemicals, biofortication and protein functionality. Journal of Cereal Science, 59,
257275.
Thompson, J. P., Brennan, P. S., Clewett, T. G., Sheedy, J. G., & Seymour, N. P. (1999).
Progress in breeding wheat for tolerance and resistance to root-lesion nematode (Pratylenchus thornei). Australasian Plant Pathology, 28, 4552.
Toktay, H., McIntyre, C. L., Nicol, J. M., Ozkan, H., & Elekcioglu, H. I. (2006). Identication of common root-lesion nematode (Pratylenchus thornei Sher et Allen) loci in bread
wheat. Genome, 49, 13191323.
Tripathi, L., Roderick, H., Babirye, A., & Atkinson, H.,J. (2014). Transgenic nematode resistance for African food security: nematode resistance bananas as a case study. In Proceeding of
the 6th international congress on nematology, 2014 (pp. 121122) (Cape Town, South Africa).
Urwin, P. E., Lilley, C. J., & Atkinson, H. J. (2002). Ingestion of double-stranded RNA by
preparasitic juvenile cyst nematodes leads to RNA interference. Molecular Plant-Microbe
Interactions, 15, 747752.
376
van der Voort, J. R., Wolters, P., Folkertsma, R., Hutten, R., Van Zandvoort, P., Vinke, H.,
et al. (1997). Mapping of the cyst nematode resistance locus Gpa2 in potato using a strategy based on comigrating AFLP markers. Theoretical and Applied Genetics, 95, 874880.
Wang, Z. H., Potter, R. H., & Jones, M. G. K. (2003). Differential display analysis of gene
expression in the cytoplasm of giant cells induced in tomato roots by Meloidogyne javanica.
Molecular Plant Pathology, 4, 361371.
Wang, Z., Shuie, L., & Jones, M. G. K. (2008). Use of a double promoter system to target
nematode feeding cells. In Proceedings of the EU COST ACTION 872 workshop, exploiting
genomics to understand plant-nematode interactions, Postonja, Slovenia, May 2008.
Ware, J. O. (1936). Plant breeding and the cotton industry. In Yearbook of agriculture (p. 657).
Washington D.C: US Department of Agriculture.
Webb, D. M., Baltazar, B. M., Rao-Arelli, A. P., Schupp, J., Clayton, K., Keim, P., et al.
(1995). Genetic mapping of soybean cyst nematode race-3 resistance loci in the soybean
PI 437.654. Theoretical and Applied Genetics, 91, 574581.
Webber, H. J., & Orton, W. A. (1902). Some diseases of cowpea. II. A cowpea resistant to
root knot nematode (Heterodera radicola). In USDA bureau of plant industry Bulletin no. 17.
Wilfarth, H. (1900). Ein neuer gesichtpunkt zur bekmpfung der nematoden. Zeitchr. d. ver
d Deut. Zucker Industrie. Lieferung, 529, 195204.
Williams, K. J., Fisher, J. M., & Langridge, P. (1996). Development of a PCR-based allelespecic assay from an RFLP probe linked to resistance to cereal cyst nematode in wheat.
Genome, 39, 798801.
Williamson, V. M., & Kumar, A. (2006). Nematode resistance in plants: the battle
underground. Trends in Genetics, 22, 396403.
Williams, K., Taylor, S., Bogacki, P., Pallotta, M., Bariana, H., & Wallwork, H. (2002).
Mapping of the root lesion nematode (Pratylenchus neglectus) resistance gene Rlnn1 in
wheat. Theoretical and Applied Genetics, 104, 874879.
Yadav, B. C., Veluthambi, K., & Subramaniam, K. (2006). Host-generated double stranded
RNA induces RNAi in plant-parasitic nematodes and protects the host from infection.
Molecular and Biochemical Parasitology, 148, 219222.
Yaghoobi, J., Kaloshian, I., Wen, Y., & Williamson, V. M. (1995). Mapping a new nematode
resistance locus in Lycopersicon peruvianum. Theoretical and Applied Genetics, 91, 457464.
Yang, Y., Jittayasothorn, Y., Chronis, D., Wang, X., Cousins, P., & Zhong, G.-Y. (2013).
Molecular characteristics and efcacy of 16D10 siRNAs in inhibiting root-knot nematode infection in transgenic grape hairy roots. PLoS One, 8, 69463. http://dx.doi.org/
10.1371/journal.pone.0069463.
Zhang, L., Hou, D., Chen, X., Li, D., Zhu, L., Zhang, Y., et al. (2012). Exogenous plant
MIR168a specically targets mammalian LDLRAP1: evidence of cross-kingdom regulation by microRNA. Cell Research, 22, 107126.
Zwart, R. S., Thompson, J. P., & Godwin, I. D. (2005). Identication of quantitative trait
loci for resistance to two species of root-lesion nematode (Pratylenchus thornei and P.
neglectus) in wheat. Crop and Pasture Science, 56, 345352.