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Cercospora leaf spot, incited by the fungus Cercospora beticola Sacc., is the most widespread and damaging sugar beet (Beta vulgaris L.) foliar disease. Under favorable for the disease agro-climatic conditions, insufficient control of epidemics leads to significant sugar yield reduction and consequent heavy economic losses. Disease control strategies basically rely on a combination of fungicide applications, growing resistant cultivars, and appropriate crop rotation. The need for a more efficient and sustainable disease management, primarily dictates the production of varieties with elevated genetic resistance and the rational use of chemical treatments to avoid the development of pathogen resistance to fungicides. Both classical and molecular breeding approaches are followed to obtain better resisting varieties, the main aims being the exploitation of additional genetic variation from natural gene pools as well as the incorporation of novel resistance traits through genetic engineering. Alternating and combining fungicides differing in mode of action, restricted use of chemicals easily provoking pathogen resistance as well as systematic risk assessment for resistance development when incorporating any novel compound, could ensure a prolonged effectiveness of the spraying programs. Designing and employing various effective integrated pest management systems is hoped to contribute towards a more effective and environmentally sound disease control.

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Salinity is one of the most severe environmental stresses, negatively affecting productivity of salt-sensitive crop species. Given that germination is the most critical phase in the plant life cycle, the present study aimed to determine seed germination potential and associated traits under salt stress conditions as a simple approach to identify salttolerant lentil genotypes.• The genetic material consisted of six lentil genotypes whose adaptation to various agroclimatic conditions is not well elucidated. Salinity stress was applied by addition of NaCl at three different levels of stress, while non-stressed plants were included as controls. Evaluation of tolerance was performed on the basis of germination percentage, seed water absorbance, root and shoot length, seedling water content, seedling vigour index and number of seedlings with an abnormal phenotype.• Overall, our findings revealed that salinity stress substantially affects all traits associated with germination and early seedling growth, with the effect of salinity being dependent on the level of stress applied. It is noteworthy, however, that genotypes responded differently to the varying salinity levels. In this context, Samos proved the most salt-tolerant genotype, indicating its possible use for cultivation under stress conditions.• In conclusion, the determination of seed germination and early growth potential may be exploited as an efficient strategy to reveal genetic variation in lentil germplasm of unknown tolerance to salinity stress. This approach allows selection of desirable genotypes at early growth stages, thus enabling more efficient application of various breeding methods to achieve stress-tolerant lentil genotypes.

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In the present work, we describe the characterisation of the glutathione transferase (GST) gene family from Agrobacterium tumefaciens C58. A genome survey revealed the presence of eight GST-like proteins in A. tumefaciens (AtuGSTs). Comparison by multiple sequence alignment generated a dendrogram revealing the phylogenetic relationships of AtuGSTs-like proteins. The beta and theta classes identified in other bacterial species are represented by five members in A. tumefaciens C58. In addition, there are three "orphan" sequences that do not fit into any previously recognised GST classes. The eight GST-like genes were cloned, expressed in Escherichia coli and their substrate specificity was determined towards 17 different substrates. The results showed that AtuGSTs catalyse a broad range of reactions, with different members of the family exhibiting quite varied substrate specificity. The 3D structures of AtuGSTs were predicted using molecular modelling. The use of comparative sequence and structural analysis of the AtuGST isoenzymes allowed us to identify local sequence and structural characteristics between different GST isoenzymes and classes. Gene expression profiling was conducted under normal culture conditions as well as under abiotic stress conditions (addition of xenobiotics, osmotic stress and cold and heat shock) to induce and monitor early stress-response mechanisms. The results reveal the constitutive expression of GSTs in A. tumefaciens and a modulation of GST activity after treatments, indicating that AtuGSTs presumably participate in a wide range of functions, many of which are important in counteracting stress conditions. These functions may be relevant to maintaining cellular homeostasis as well as in the direct detoxification of toxic compounds.

To explore possible sources of transgenic resistance to the rhizomania-causing Beet necrotic yellow vein virus (BNYVV), Nicotiana benthamiana plants were constructed to express the harpin of Pseudomonas syringae pv. phaseolicola (HrpZPsph). The HrpZ protein was expressed as an N-terminal fusion to the PR1 signal peptide (SP/HrpZ) to direct harpin accumulation to the plant apoplast. Transgene integration was verified by mPCR in all primary transformants (T0), while immunoblot analysis confirmed that the protein HrpZPsph was produced and the signal peptide was properly processed. Neither T0 plants nor selfed progeny (T1) showed macroscopically visible necrosis or any other macroscopic phenotypes. However, plants expressing the SP/HrpZPsph showed increased vigor and grew faster in comparison with non-transgenic control plants. Transgenic resistance was assessed after challenge inoculation with BNYVV on T1 progeny by scoring of disease symptoms and by DAS-ELISA at 20 and 30 dpi. Transgenic and control lines showed significant differences in terms of the number of plants that became infected, the timing of infection and the disease symptoms displayed. Plants expressing the SP/HrpZPsph developed localized leaf necrosis in the infection area and had enhanced resistance upon challenge with BNYVV. In order to evaluate the SP/HrpZ-based resistance in the sugar beet host, A. rhizogenes-mediated root transformation was exploited as a transgene expression platform. Upon BNYVV inoculation, transgenic sugar beet hairy roots showed high level of BNYVV resistance. In contrast, the aerial non-transgenic parts of the same seedlings had virus titers that were comparable to those of the seedlings that were untransformed or transformed with wild type R1000 cells. These findings indicate that the transgenically expressed SP/HrpZ protein results in enhanced rhizomania resistance both in a model plant and sugar beet, the natural host of BNYVV. Possible molecular mechanisms underlying the enhanced resistance and plant growth phenotypes observed in SP/HrpZ transgenic plants are discussed.

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