Christos Dordas

Role of Nutrients in Controlling Plant Diseases in Sustainable Agriculture: A Review

In recent years the importance of sustainable agriculture has risen to become one of the most important issues in agriculture. In addition, plant diseases continue to play a major limiting role in agricultural production. The control of plant diseases using classical pesticides raises serious concerns about food safety, environmental quality and pesticide resistance, which have dictated the need for alternative pest management techniques. In particular, nutrients could affect the disease tolerance or resistance of plants to pathogens. However, there are contradictory reports about the effect of nutrients on plant diseases and many factors that influence this response are not well understood. This review article summarizes the most recent information regarding the effect of nutrients, such as N, K, P, Mn, Zn, B, Cl and Si, on disease resistance and tolerance and their use in sustainable agriculture. There is a difference in the response of obligate parasites to N supply, as when there is a high N level there is an increase in severity of the infection. In contrast, in facultative parasites at high N supply there is a decrease in the severity of the infection. K decreases the susceptibility of host plants up to the optimal level for growth and beyond this point there is no further increase in resistance. In contrast to K, the role of P in resistance is variable and seemingly inconsistent. Among the micronutrients, Mn can control a number of diseases as Mn has an important role in lignin biosynthesis, phenol biosynthesis, photosynthesis and several other functions. Zn was found to have a number of different effects as in some cases it decreased, in others increased, and in others had no effect on plant susceptibility to disease. B was found to reduce the severity of many diseases because of the function that B has on cell wall structure, plant membranes and plant metabolism. Cl application can enhance host plants’ resistance to disease. Si has been shown to control a number of diseases and it is believed that Si creates a physical barrier which can restrict fungal hyphae penetration, or it may induce accumulation of antifungal compounds. Integrative plant nutrition is an essential component in sustainable agriculture, because in most cases it is more cost-effective and also environmentally friendly to control plant disease with the adequate amount of nutrients and with no pesticides. Nutrients can reduce disease to an acceptable level, or at least to a level at which further control by other cultural practices or conventional organic biocides are more successful and less expensive.

Evidence for channel mediated transport of boric acid in squash (Cucurbita pepo)

Boron (B) is taken up by plant roots as undissociated boric acid which is a non-electrolyte of similar size to urea and other non-electrolytes. In animal systems, non-electrolytes are transported across biological membranes through aquaporins or through non-aquaporin channels. In artificial lipids boric acid is known to diffuse directly through the lipid bilayer at a rate that is determined by lipid composition. A possible role for channel proteins in in-vitro B uptake is suggested by recent work in which B uptake into isolated membrane vesicles was inhibited by channel blockers and by demonstration that the expression of the plant channel protein PIP1 in Xenopus oocytes increases boric acid uptake by 30%. This study examines whether B transport is a channel-mediated process in intact plants. In the presence of the channel inhibitors HgCl2, phloretin, and DIDS, B uptake by squash plants was reduced by 40–90% by HgCl2 (as HgCl2 varied from 50 μM to 1 mM), 44% by phloretin (250 μM) and 58% by DIDS (250 μM). The effect of Hg ions on B uptake was reversed by 2-mercaptoethanol. The addition of other non-electrolytes in size ranges similar to boric acid inhibited B uptake to various degrees. Addition of urea resulted in 54% decrease in B uptake, while, acetamide, formamide, thiourea and glycerol inhibited uptake by 50, 35, 53 and 44%, respectively. The effect of HgCl2 on B uptake was greater at high B concentrations than at low B concentrations. These data and information from in-vivo studies suggest two possible mechanisms of B uptake: passive diffusion through lipid bilayers and channel-mediated transport.

Nonsymbiotic hemoglobins and stress tolerance in plants.

Hemoglobins (Hbs) are heme containing proteins found in most organisms including animals, bacteria, and plants. Their structure, size, and function are quite diverse among the different organisms. There are three different types of hemoglobins in plants: symbiotic (sHb), nonsymbiotic (nsHb), and truncated hemoglobins (trHb). The nonsymbiotic hemoglobins are divided into: class 1 hemoglobins (nsHb-1s), which have a very high affinity for oxygen: and class 2 hemoglobins (nsHb-2s), which have lower affinity for oxygen, are similar to the sHbs. nsHb-1s are expressed under hypoxia, osmotic stress, nutrient deprivation, cold stress, rhizobial infection, nitric oxide exposure, and fungal infection. Tolerance to stress is very important for the survival of the plant. Hemoglobins are one of many different strategies that plants have evolved to overcome stress conditions and survive. Hbs also react with NO produced under different stress conditions. Class 1 nsHbs are involved in a metabolic pathway involving NO. Those hemoglobins provide an alternative type of respiration to mitochondrial electron transport under limiting oxygen concentrations. Class 1 nsHbs in hypoxic plants act as part of a soluble, terminal, NO dioxygenase system, yielding nitrate from the reaction of oxyHb with NO. The overall reaction sequence, referred to as the nsHb/NO cycle, consumes NADH and maintains ATP levels via an as yet unknown mechanism. Class 2 nsHbs seem to scavenge NO in a similar fashion as class 1 Hbs and are involved in reducing flowering time in Arabidopsis. nsHbs also show peroxidase-like activity and NO metabolism and possibly protect against nitrosative stress in plant-pathogen interaction and in symbiotic interactions. nsHbs may be involved in other stress conditions such as osmotic, nutrient and cold stress together with NO and the function of nsHbs can be in NO metabolism and signal transduction. However, other possible functions cannot be precluded as Hbs have many different functions in other organisms.

Sustainable production of barley and wheat by intercropping common vetch

Intercropping legumes with cereals for forage production is a sustainable technique showing several environmental benefits. We studied yields, quality and the growth rate of a 2-year experiment including (1) sole crops of common vetch, barley and winter wheat, and (2) intercrops of common vetch with barley and winter wheat using seeding ratios of 55:45 and 65:35. Our results show that the greatest dry matter yields were obtained with wheat and barley sole crops. The lowest yield was obtained with common vetch sole crop. The intercrops produced about 13–30% more dry matter than the common vetch sole crop, but 12–23% less than cereal sole crops. Further, the growth rate of common vetch and cereals was greater when species were grown as sole crops than in intercrops. Quality components indicated an advantage for the sole crop of common vetch followed by its intercrop with barley at the 65:35 seeding ratio, which exhibited higher crude protein yield than the sole crop of cereals and the other intercrops. The results of this study indicate that common vetch intercrops with barley or winter wheat produced higher dry matter than common vetch sole crop. In addition, the intercrop of common vetch with barley (65:35) provided higher forage quality than the other intercrops. Our study highlights that vetch-cereal intercropping can be used as an alternative cropping system which combines sustainability due to N fixation from common vetch, and high yield and forage quality.

Boron deficiency affects cell viability, phenolic leakage and oxidative burst in rose cell cultures

Despite the fact that the effect of B deficiency on cell metabolism has been studied extensively the mechanism by which B deficiency causes cell death has not been determined. Several authors have hypothesized that B deficiency leads to oxidative burst and hence cell death, though this has not been demonstrated experimentally. In the present work we utilize rose cell (Rosa damascena Mill cv Gloide de Guilan) suspension culture, maintained at the stationary growth phase to determine the effect of B deficiency on cell viability and a number of physiological and biochemical parameters including H2O2 production, phenolic leakage, pH of the medium, B concentration and biomass. B deficiency resulted in the death of some cells as early as 24 h following B deprivation, and continued rapidly in the following days. In B deficient cells a small oxidative burst (indicated by the production of H2O2) was observed coincident with first cell death and increasing thereafter. Increasing amounts of phenolics were observed in the culture medium of the deficient treatment indicating loss of membrane integrity, however results suggest this increase is a secondary consequence of cell death. The effect of B deficiency on the oxidative burst, together with the effect on cell viability is discussed.

Boron application affects seed yield and seed quality of sugar beets

Sugar beet (Beta vulgaris L.) is one of the most important sugar crops worldwide. Despite the fact that sugar beet crop has high requirements for boron (B), the effect of B applications on seed yield and on seed quality is not known. A 2-year field study was conducted to determine whether soil and foliar B applications during anthesis increase seed set, final seed yield and improve seed quality of sugar beets. Boron solutions were applied at four rates (0, 245, 490 and 735 mg/l of B) as foliar applications and at two rates (1·5 and 3 kg/ha of B) as soil applications to field plots exhibiting no vegetative symptoms of B deficiency. Foliar B application increased the concentration of B in vegetative and reproductive tissues much more than soil application. In addition, foliar B application increased the seed yield by an average of 10 % in the first year and by an average of 44% in the second year. The mean seed weight was affected by B application as it was increased in both years. The proportion of larger seeds (>5·00 and 4·5-5·00 mm) increased with increasing application of B. Moreover, seed quality was affected and the proportion of abnormal seedlings was decreased with B application. However, seed vigour was not affected by B application. These data indicate that foliar B application can improve seed yield and seed quality of sugar beet grown for seed production. However, the physiological basis of this effect remains unknown.

Corn poppy (Papaver rhoeas) cross-resistance to ALS-inhibiting herbicides.

BACKGROUND Papaver rhoeas (L.) has evolved resistance to tribenuron in winter wheat fields in northern Greece owing to multiple Pro(197) substitutions. Therefore, the cross-resistance pattern to other sulfonylurea and non-sulfonylurea ALS-inhibiting herbicides of the tribenuron resistant (R) and susceptible (S) corn poppy populations was studied by using whole-plant trials and in vitro ALS catalytic activity assays. RESULTS The whole-plant trials revealed that tribenuron R populations were also cross-resistant to sulfonylureas mesosulfuron + iodosulfuron, chlorsulfuron and triasulfuron. The whole-plant resistance factors (RFs) calculated for pyrithiobac, imazamox and florasulam ranged from 12.4 to > 88, from 1.5 to 28.3 and from 5.6 to 25.4, respectively, and were lower than the respective tribenuron RF values (137 to > 2400). The ALS activity assay showed higher resistance of the ALS enzyme to sulfonylurea herbicides (tribenuron > chlorsulfuron) and lower resistance to non-sulfonylurea ALS-inhibiting herbicides (pyrithiobac > florasulam ≈ imazamox). CONCLUSION These findings indicate that Pro(197) substitution by Ala, Ser, Arg or Thr in corn poppy results in a less sensitive ALS enzyme to sulfonylurea herbicides than to other ALS-inhibiting herbicides. The continued use of sulfonylurea herbicides led to cross-resistance to all ALS-inhibiting herbicides, making their use impossible in corn poppy resistance management programmes.

Multiple Pro197 Substitutions in the Acetolactate Synthase of Corn Poppy (Papaver rhoeas) Confer Resistance to Tribenuron

Abstract Variations in the acetolactate synthase (ALS) gene sequence were determined from 28 populations of corn poppy resistant (R) to tribenuron and from 6 populations susceptible (S) to this herbicide. The ALS gene fragment (634 bp) sequence revealed in R populations five point mutations at the codon Pro197, and among them the substitution of Pro197 by Ala was the most common. The sequencing chromatograms revealed that nine R individuals had only the mutant ALS gene and were homozygous (RR), 18 R individuals had both the wild type and the mutant ALS gene and were heterozygous (RS), whereas one R individual was heterozygous but with two different mutant ALS alleles (R1R2). The use of restriction digestion profile analysis to verify the DNA sequence results by detecting the existence of point mutations at the codon 197 managed to distinguish the R and S alleles and confirmed the results obtained from the sequencing chromatograms analysis. The secondary protein structure prediction suggested the formation of novel β-strands for each of the five mentioned amino acid substitutions that was not present in wild type ALS around the mutant site. These findings support the hypothesis that the substitution of Pro197 by Ser, Thr, Ala, Arg, or Leu resulted in altered secondary structure, which stabilizes an ALS tertiary conformation that prevents tribenuron binding and thus confers resistance to this herbicide. Nomenclature: Tribenuron; corn poppy, Papaver rhoeas L. PAPRH.

INCREASED CONCENTRATION OF SOIL CADMIUM AFFECTS ON PLANT GROWTH, DRY MATTER ACCUMULATION, Cd, AND Zn UPTAKE OF DIFFERENT TOBACCO CULTIVARS (NICOTIANA TABACUM L.)

Cadmium (Cd) is a toxic trace metal pollutant for humans, animals, and plants. Tobacco is a wellknown efficient accumulator of Cd and the genotypic differences in Cd uptake and the response to Cd was not determined. The objectives of this study were to investigate: 1) the effects of Cd on the growth and development of different tobacco cultivars; 2) the differences among tobacco cultivars in Cd concentration, uptake, and use for the phytoremediation of polluted soils with Cd; and (3) the interactions between Cd and Zn with respect to concentration and uptake. The Cd level affected the number of leaves and dry matter accumulation, and there were differences among the different cultivars that were used. Furthermore, some cultivars showed a higher reduction in growth than others, indicating that they are more sensitive to Cd level in the soil. Moreover, differences existed among the cultivars for the Cd concentration and uptake. There also were negative correlations between Cd and Zn concentrations; as Cd accumulation increased, Zn accumulation decreased, which showed that the two heavy metals were antagonistic. These results suggest that tobacco cultivars differed greatly in their growth and developmental responses to Cd and in the concentration and uptake of Cd and Zn. In addition, it is possible to use certain tobacco cultivars to lower the Cd concentration in the soil.

Permeability and the mechanism of transport of boric acid across the plasma membrane of Xenopus laevis oocytes.

Boron is an essential element for vascular plants and for diatoms, cyanobacteria, and a number of species of marine algal flagellates. Boron was recently established as an essential micronutrient for frogs (Xenopus laevis) and preliminary evidence suggests that it may be essential for all animals. The main form of B, which is available in the natural environment, is in the form of undissociated boric acid. The permeability coefficient and the mechanism of transport of boric acid, however, have not been experimentally determined across any animal membrane or cell. In the experiments described here, the permeability coefficient of boric acid in Xenopus oocytes was 1.5 × 10−6 cm/s, which is very close with the permeability across liposomes made with phosphatidylcholine and cholesterol (the major lipids in the oocyte membrane). Moreover, we investigated the mechanism of boric acid movement across the membrane of Xenopus oocytes and we compared it with the transport across artificial liposomes. The transport of boric acid across Xenopus oocytes was not affected by inhibitors such as HgCl2, phloretin, or 4,4-diisothiocyanatostilbene-2,2′-d-sulfonic acid (DIDS). The kinetics of B uptake was linear with concentration changes, and the permeability remained the same at different external boric acid concentrations. These results suggest that B transport occurs via simple passive diffusion through the lipid bilayer in Xenopus oocytes.