Joint research project

Cicer arietinum ancient varieties in Egypt: exploiting associated pathogens and modern defence strategies.

Project leaders
Vitantonio Pantaleo, Moemen Sayed Hanafy
Agreement
EGITTO - NRC - National Research Centre of Egypt
Call
CNR-NRC 2016-2017
Department
Biology, agriculture and food sciences
Thematic area
Biology, agriculture and food sciences
Status of the project
New

Research proposal

In many countries, grain legumes have gained much importance in view of the acute shortage in the production of animal proteins and the prevalence of protein malnutrition. This makes the grain legumes to be considered as the "poor men's meat in the developing countries but also a valid alternative of protein source for specific (vegetarian or vegan) or balanced diets in developed countries.

With a worldwide production of 73x106 (FAOSTAT 2013) tons, grain legumes (also known as pulses) rank third after cereals and oilseeds. Moreover, legumes are able to fix the atmospheric nitrogen in association with bacteria and play a central role in low input (Sustainable Agricultural Systems) (Graham and Vance 2003). The world production of chickpea on 2013 was 13x106 tones (FAOSTAT 2013), makes chickpea rank fourth among the pulses after soybean, peanut and common bean. However, chickpea can be the most important crop at a regional level, especially in semi-arid areas of the world (Popelka and Higgins 2007) and in Mediterranean regions.
The seeds of chickpea contains about 20% protein, about 55% carbohydrate and about 5% fat and represent a basic food crop in many developing countries. The productivity of chickpea has been limited due to their susceptibility to viruses, pathogens and pests, sensitivity to environmental stresses and poor pollination. Commercially, the species is grouped into the desi-type (small-seeded) chickpea that accounts for about 85% of world production (7.8×106 t) and is the principal type grown in India, Pakistan, Iran, Afghanistan and Ethiopia. However, the Kabuli-type (large seeded) is grown in the Middle East, India, Mexico as well as in North America, Australia and Spain (Popelka and Higgins 2007). The colour of the seeds (black, red or white) is also a key commercial character and it is associated to the content of antioxidants, that are molecules relevant for human health. The genetic improvement of chickpea by conventional breeding has been met with low success due to the narrow genetic base and sexual incompatibility for interspecific hybridization (Somers et al. 2003; Popelka et al. 2004). Therefore, biotechnological approaches offer a visible strategy to improve the grain productivity of this important legume.

Most legumes, especially large-seeds grain legumes such as chickpea, faba bean, pigeonpea and phaseolus, are considered recalcitrant to in vitro regeneration and transformation (Link et al. 2008; Hanafy et al. 2008; Varshney et al. 2009). Therefore, development of a reliable genetic transformation technique will be of great importance to incorporate the desirable traits into chickpea. The number of reports of successful genetic transformation of chickpea is relatively low, however the NRC labs possess all the necessary skills to develop C. arietinum transgenic plants


Chickpea chlorotic dwarf virus (CCDV) is a geminivirus transmitted by the leafhopper Orosius orientalis; it infects tobacco species and grain legumes including chickpea (Horn et al., 1993). The symptoms developed following CCDV infection include plant stunting, internode shortening, phloem browning in the collar region, with leaf reddening in desi-type or yellowing in kabuli-type chickpeas until severe yield losses (Horn et al., 1993).
Viral infections in legumes may also interfere with the seed colour . Indeed, viral silencing suppressors are known to subvert post-trasnscriptional regulation of endogenous genes, including those involved in antioxidant accumulation and colour of the seed (Senda et al., 2014).
Chickpea may also be infected by phytoplasmas, phloematic bacteria transmitted by phloem feeder hemipteran insects. Different ribosomal sub-groups within the 'Candidatus Phytoplasma aurantifolia' species have been associated to dwarfing and phyllody symptoms in this species in several parts of the world (Chen et al., 2011). Phytoplasma infection hampers pod production, and infected plants are a phytoplasma reservoir for potential vectors to other important crops such as citrus.
Finally, plant-parasitic nematodes can significantly limit chickpea production worldwide (Castillo et al., 2008). Moreover, nematode attacks can increase the sensitivity of the infected plants to other biotic and abiotic stresses with resulting stunting and poor yield (Sikora et al 2005). The most aggressive parasites of chickpea are cyst-forming nematodes (Heterodera spp.) microscopic roundworm reported on roots of many legume crops. They causes very important losses annually; therefore, nematode control is an important goal in the development of sustainable chickpea production systems. The chemical control is not sustainable due to economic and environmental reasons. The gene SHMT (Li et al., 2012) can have wide-ranging effects on one-carbon folate metabolism and alterations of the folate homeostasis leading to folate deficiency may trigger hypersensitive-response-like programmed cell death of the developing feeding cells and subsequent death of nematode.
Alternatively, in resistant cultivar the gene may lead to the production of nematicidal compound or serve as the target of a nematode-secreted small molecule or effector protein to trigger a resistance signalling pathway

Research goals

With the present proposal we intend to launch a combined and collaborative effort aiming to improve existing protocols for in vitro plant regeneration and genetic transformation systems of Egyptian local chickpea cultivars (Dr. M. Hanafy and Co). The ultimate aim of such Project is the development of meaningful CCDV and nematode resistant breeding lines (Dr. E. Noris and Dr. P. Leonetti, respectively). Moreover, specific transgenic plants will also be useful tools for the identification of genes involved in the production of antioxidants in the seeds, which are controlled at post-transcriptional levels by short non-coding RNAs (Dr. V. Pantaleo). Selected Egyptian varieties will also be screened for possible novel phloematic bacteria that could be relevant as emerging plant diseases in the Mediterranean basin (Dr. C. Marzachì).
The objectives of the proposal are summarized herebelow:

1) Improving the transformation protocol for the selected Egyptian cultivars;
2) Obtain resistant chickpea plants capable to reduce the yield loses caused by CCDV and cyst nematodes, thus improving the food security and self-sufficiency in Egypt;
3) Obtain transgenic chickpea plants expressing a selected viral suppressors able to subvert the Chalcone syntase pathway;
4) Screening for novel phloematic bacteria hosted by chickpea;
5) Transferring the European experience in molecular breeding and genetic engineering to Egypt through continuing our close cooperation between NRC-Egypt and CNR-Italy.

Last update: 28/03/2024