Changes in physiology and biochemistry of mottle streak virus infected finger millet plants

A significant reduction in the growth parameters viz., plant height, number of tillers, number of productive tillers, leaves, leaf area, 1000 grain weight and grain yield were observed in the mottle streak virus infected finger millet plants compared to healthy finger millet plants. The germination and vigour of seedlings from the seeds of infected plants were reduced. Physiological changes in finger millet as a result of virus infection were investigated. The chlorophyll pigments ‘a’ and ‘b’ as well as total chlorophyll were reduced due to mottle streak infection. The virus infection led to increased total sugar, starch, soluble protein and phenol contents. The mineral metabolism of infected plants showed a reduction in nitrogen, phosphorus, potassium, magnesium, calcium and iron.     

The Effects of Co-Infection by Sunn-Hemp Mosaic Virus (SHMV) and Fusarium oxysporum on the Growth of French Bean

In the seedlings of three cultivars of French bean (Phaseolus vulgaris), ‘Prince,’‘Masterpiece’ and ‘Pinto’, co-infection by Sunn-hemp mosaic virus (SHMV) with either of the vascular wilt pathogens, Fusarium oxysporum f. sp. phaseoh (Fop) or f. sp. tracbeiphilum (Fot) caused greater losses in total fresh weight and in leaf area compared with uninfected plants or plants infected singly with any one of these pathogens. Co-infection of a fourth cultivar (‘Canadian Wonder’) had no greater effects on growth reduction than single infection. The concentration of SHMV in the leaves of ‘Prince’ and to a greater extent in the leaves of ‘Masterpiece’ increased more after double infection than with infection by the virus alone. The nature and possible mechanisms of the pathogenic effects in French bean are discussed.     

Flavescence dorée phytoplasma deregulates stomatal control of photosynthesis in Vitis vinifera

Flavescence dorée (FD) is among the major grapevine diseases causing high management costs; curative methods against FD are unavailable. In FD‐infected plants, decrease in photosynthesis is usually recorded, but deregulation in stomatal control of leaf gas exchange during FD infection and recovery is unknown. We measured the seasonal time course of gas exchange rates in two cultivars (‘Barbera’ and ‘Nebbiolo’) during the term of 1 year when grapevines experienced a water stress and another with no drought, with difference in gas exchange rates in response to FD infection and recovery as assessed by symptom observation and phytoplasma detection through PCR analysis. Chlorophyll fluorescence was also evaluated at the time of maximum symptom severity in ‘Barbera’, the cultivar showing the most severe stress response to FD infection, causing the highest damage in vineyards of north‐western Italy. In FD‐infected plants, net photosynthesis and transpiration gradually decreased during the season, more during the no drought year than during drought. During recovery, healthy (PCR negative) plants infected 2 years before, but not those infected an year before, regained the gas exchange performances to the level as measured before infection. The relationships between stomatal conductance and the residual leaf intercellular CO₂ concentration (c_i) discriminated healthy versus FD‐infected and recovered plants; at the same c_i, FD‐infected leaves had higher non‐photochemical quenching than healthy ones. We conclude that metabolic, not stomatal, leaf gas exchange limitation in FD‐infected and recovered grapevines is the basis of plant response to FD disease. In addition, we also suggest that such response is dependent upon water stress, by showing that water stress superimposes on FD infection in terms of stomatal and metabolic non‐stomatal limitations to carbon assimilation.     

Aphid behavioral responses to virus‐infected plants are similar despite divergent fitness effects

We compared the settling preferences and reproductive potential of an oligophagous herbivore, the pea aphid, Acyrthosiphon pisum Harris (Hemiptera: Aphididae), in response to pea plants, Pisum sativum L. cv. ‘Aragorn’ (Fabaceae), infected with two persistently transmitted viruses, Pea enation mosaic virus (PEMV) and Bean leaf roll virus (BLRV), that differ in their distribution within an infected plant. Aphids preferentially oriented toward and settled on plants infected with PEMV or BLRV in comparison with sham‐inoculated plants (plants exposed to herbivory by uninfected aphids), but aphids did not discriminate between plants infected with the two viruses. Analysis of plant volatiles indicated that plants inoculated with either virus had significantly higher green leaf volatile‐to‐monoterpene ratios. Time until reproductive maturity was marginally influenced by plant infection status, with a trend toward earlier nymph production on infected plants. There were consistent age‐specific effects of plant infection status on aphid fecundity: reproduction was significantly enhanced for aphids on BLRV‐infected plants across most time intervals, though mean aphid fecundity did not differ between sham and PEMV‐infected plants. There was no clear pattern of age‐specific survivorship; however, mean aphid lifespan was reduced on plants infected with PEMV. Our results are consistent with predictions of the host manipulation hypothesis, extended to include plant viruses: non‐viruliferous A. pisum preferentially orient to virus‐infected host plants, potentially facilitating pathogen transmission. These studies extend the scope of the host manipulation hypothesis by demonstrating that divergent fitness effects on vectors arise relative to the mode of virus transmission.     

Heavy metal soil contamination delays the appearance of virus-induced symptoms on potato but favours virus accumulation

Abstract

Here we report on the investigation of virus infection development in plants undergoing simultaneous heavy metal stress. We carried out small-scale field experiments using model system Potato virus X (PVX) – Solanum tuberosum cv. Povin' (potato) plants. Heavy metals zinc (Zn), copper (Cu) and lead (Pb) were added to the soil separately (monometal contamination) at a range of concentrations. Our results show that heavy metal stress significantly delays the appearance and potentiates severity of virus-specific symptoms on infected potato plants. We also demonstrate that PVX content in plants may increase tremendously in response to this abiotic stressor. Finally, we provide links to the possible consequences in the context of virus epidemiology.     

The effect of cassava mosaic geminiviruses on symptom severity, growth and root yield of a cassava mosaic virus disease-susceptible cultivar in Uganda

A study was carried out to assess the effect of different cassava mosaic geminiviruses (CMGs) occurring in Uganda on the growth and yield of the susceptible local cultivar ‘Ebwanateraka’. Plants infected with African cassava mosaic virus (ACMV), ‘mild’ and ‘severe’ strains of East African cassava mosaic virus‐Uganda (EACMV‐UG2) and both ACMV and EACMV‐UG2 were grown in two experiments in Kabula, Lyantonde in western Uganda. The most severe disease developed in plants co‐infected with ACMV and EACMV‐UG2 and in those infected with the ‘severe’ form of EACMV‐UG2 alone; disease was least severe in plants infected with the ‘mild’ strain of EACMV‐UG2. ACMV‐infected plants and those infected with the ‘mild’ strain of EACMV‐UG2 were tallest in the 1999–2000 and 2000–2001 trials, respectively; plants dually infected with ACMV and EACMV‐UG2 were shortest in both trials. Plants infected with ‘mild’ EACMV‐UG2 yielded the largest number and the heaviest tuberous roots followed by ACMV and EACMV‐UG2 ‘severe’, respectively, whilst plants dually infected with ACMV and EACMV‐UG2 yielded the least considering the two trials together. Reduction in tuberous root weight was greatest in plants dually infected with ACMV and EACMV‐UG2, averaging 82%. Losses attributed to ACMV alone, EACMV‐UG2 ‘mild’ and EACMV‐UG2 ‘severe’ were 42%, 12% and 68%, respectively. Fifty percent and 48% of the plants infected with both ACMV and EACMV‐UG2 gave no root yield in 1999–2000 and 2000–2001, respectively. These results indicate that CMGs, whether in single or mixed infections, reduce root yield and numbers of tuberous roots produced and that losses are substantially increased following mixed infection.     

Effect of different manuring and defoliation patterns on broad-leaved dock (Rumex obtusifolius) in grassland

The effects of cutting frequency and cutting height on broad‐leaved dock (Rumex obtusifolius) in Lolium perenne‐based agricultural grassland at two levels of fertiliser input were investigated at North Wyke, Devon, UK. Two micro‐plot field experiments, containing immature dock plants at uniform densities, with a factorial design, were used to compare: (a) ‘organic’ and ‘low‐input’ fertilisation, i.e. cattle slurry only vs slurry plus mineral fertiliser (NPK at 100‐0‐64 kg ha^?1 yr^?1), (b) cutting heights of 5–6 cm vs 10–12 cm, and (c) four harvesting frequencies representative of different grassland management practices (regular 4‐weekly cutting, a ‘hay‐stage’ cutting, and two treatments with ‘silage‐stage’ cutting). Expt 1 was established in 1995 with 13 dock plants m^?2 (from excised dock‐root sections) and Expt 2 established in 1996 with 25 plants m^?2 (as seed‐grown plug plants). Treatments were assessed over the 2 subsequent years to determine treatment effects on total herbage dry matter (DM) yield and dock DM yield, and on in‐situ measures of dock ramets. In both experiments, total DM yield was increased by 1.0–2.01 ha^?1 yr^?1 for treatments receiving NK fertiliser; the proportion of dock was also higher than from slurry‐only treatments. In Expt 1, the dock ramet density, mean dock ramet height, mean leaf length and numbers of dock leaves per m² were also greater on NK fertilised treatments in autumn of yr 1. Height of cut had no consistent effect on dock yield, but dock ramet density and leaf density in autumn were greater on the 5–6 cm than the 10–12 cm cutting treatment, Expt 2 only. In yr 2 of both experiments cutting at 4‐weekly intervals resulted in less dock in the herbage than hay‐stage cutting and, particularly in Expt 1, there were associated differences in leaf density and ramet height in autumn; silage‐stage treatments were intermediate. Results are discussed in relation to requirements for management options where there is a need to avoid or reduce herbicides.     

Excising stem samples underwater at native tension does not induce xylem cavitation

Xylem resistance to water stress‐induced cavitation is an important trait that is associated with drought tolerance of plants. The level of xylem cavitation experienced by a plant is often assessed as the percentage loss in conductivity (PLC) at different water potentials. Such measurements are constructed with samples that are excised underwater at native tensions. However, a recent study concluded that cutting conduits under significant tension induced cavitation, even when samples were held underwater during cutting. This resulted in artificially increased PLC because of what we have termed a ‘tension‐cutting artefact’. We tested the hypothesized tension‐cutting artefact on five species by measuring PLC at native tension compared with after xylem tensions had been relaxed. Our results did not support the tension‐cutting artefact hypothesis, as no differences were observed between native and relaxed samples in four of five species. In a fifth species (Laurus nobilis), differences between native and relaxed samples appear to be due to vessel refilling rather than a tension‐cutting effect. We avoided the tension‐cutting artefact by cutting samples to slightly longer than their measurement length and subsequent trimming of at least 0.5 cm of sample ends prior to measurement.     

Trypsin inhibitor activity in mature tobacco and tomato plants is mainly induced locally in response to insect attack,wounding and virus infection

Wounding of plants by insects is often mimicked in the laboratory by mechanical means such as cutting or crushing, and has not been compared directly with other forms of biotic stress such as virus infection. To compare the response of plants to these types of biotic and abiotic stress, trypsin inhibitor (TI) activity induced locally and systemically in mature tobacco (Nicotiana tabacum L.) and tomato (Lycopersicon esculentum L.) plants was followed for 12 days. In tobacco, cutting, crushing and insect feeding all induced comparable levels of TI activity of approx. 5 nmol·(mg leaf protein)^?1 in wounded leaves, while tobacco mosaic virus (TMV) infection of tobacco induced 10-fold lower amounts in the infected leaves. In tomato, feeding by insects also led to the induction of a level of TI activity of 5 nmol·(mg leaf protein)^?1. In contrast, both cutting and crushing of tomato leaves induced 10-fold higher amounts. These data show that biotic stress, in the form of insect feeding and TMV infection, and abiotic stress, in the form of wounding, have different effects on local levels of induced TI activity in mature tobacco and tomato plants. Irrespective of the type of wounding, in neither tobacco nor tomato could systemic induction of TI activity be observed in nearby unwounded leaves, which suggests that systemic induction of TI activity in mature tobacco and tomato plants is different from systemic TI induction in seedlings. Wounding of tobacco leaves, however, did increase the responsiveness to wounding elsewhere in the plant, as measured by an increased induction of TI activity.     

Comparison of the potential rate of population increase of brown and green color morphs of Sitobion avenae (Homoptera: Aphididae) on barley infected and uninfected with Barley yellow dwarf virus

Life tables of brown and green color morphs of the English grain aphid, Sitobion avenae (Fabricius) reared on barley under laboratory conditions at 20 ± 1°C, 65% ± 5% relative humidity and a photoperiod of 16 : 8 h (L : D) were compared. The plants were either: (i) infected with the Barley yellow dwarf virus (BYDV); (ii) not infected with virus but previously infested with aphids; or (iii) healthy barley plants, which were not previously infested with aphids. Generally, both color morphs of S. avenae performed significantly better when fed on BYDV‐infected plants than on plants that were virus free but had either not been or had been previously infested with aphids. Furthermore, when fed on BYDV‐infected plants, green S. avenae developed significantly faster and had a significantly shorter reproductive period than the brown color morph. There were no significant differences in this respect between the two color morphs of S. avenae when they were reared on virus‐free plants that either had been or not been previously infested with aphids. These results indicate that barley infected with BYDV is a more favorable host plant than uninfected barley for both the color morphs of S. avenae tested, particularly the green color morph.     

Comparison of environmental sampling methods for detecting Salmonella in commercial laying flocks in the UK

Aims: To investigate the performance of the Salmonella National Control Programme (NCP) sampling/testing methods in laying flocks of domestic fowl. Methods and Results: Eighty‐five visits were made to 69 flocks representative of the main production systems (cage, barn and free‐range) infected with Salmonella. In each visit, three methodologies were compared: (i) the European Union (EU) baseline survey method (five faeces and two dust samples); (ii) an in‐house (Veterinary Laboratories Agency, VLA) ‘wet’ method that involved collecting 10 dust and 10 faeces samples into jars with buffered peptone water; and (iii) a method involving two samples of pooled faeces and one of dust (cultured as one sample of each type), which has been adopted for the NCP for laying flocks across the EU. Conclusions: The ‘wet’ method was the most sensitive, and the NCP the least, although individual NCP samples were the most sensitive ones. Significance and Impact of the Study: The apparent lower sensitivity of the NCP method may be compensated by repeated sampling of flocks (twice during rear and several times during lay). Sampling using VLA methodology should be advocated for farms aiming to disclose low‐level Salmonella before restrictions on the sale of eggs from Salmonella Enteritidis or Salmonella Typhimurium‐infected flocks are in place.     

Age-related Resistance in Bell Pepper to Cucumber mosaic virus

We demonstrated the occurrence of mature plant resistance in Capsicum annuum‘Early Calwonder’ to Cucumber mosaic virus (CMV) under greenhouse conditions. When Early Calwonder plants were sown at 10 day intervals and transplanted to 10‐cm square pots, three distinct plant sizes were identified that were designated small, medium and large. Trials conducted during each season showed that CMV accumulated in inoculated leaves of all plants of each size category. All small plants (with the exception of the winter trial) developed a systemic infection that included accumulation of CMV in uninoculated leaves and severe systemic symptoms. Medium plants had a range of responses that included no systemic infection to detection of CMV in uninoculated leaves with the systemically infected plants being either symptomless or expressing only mild symptoms. None of the large plants contained detectable amounts of CMV in uninoculated leaves or developed symptoms. When plants were challenged by inoculation of leaves positioned at different locations along the stem or different numbers of leaves were inoculated, large plants continued to accumulate CMV in inoculated leaves but no systemic infection was observed. When systemic infection of large plants did occur, e.g. when CMV‐infected pepper was used as a source of inoculum, virus accumulation in uninoculated leaves was relatively low and plants remained symptomless. A time‐course study of CMV accumulation in inoculated leaves revealed no difference between small and large plants. Analyses to examine movement of CMV into the petiole of inoculated leaves and throughout the stem showed a range in the extent of infection. While all large plants contained CMV in inoculated leaves, some had no detectable amounts of virus beyond the leaf blade, whereas others contained virus throughout the length of the stem but with limited accumulation relative to controls.     

Plant water stress and previous herbivore damage affect insect performance

1. Short‐term changes in plant resistance traits can be affected by abiotic factors or damage by herbivores, although how the combined effects of abiotic factors and previous damage affect subsequent insect larval development is not well understood. 2. Complementary glasshouse and field experiments were conducted to evaluate whether plant water stress and previous herbivore damage influenced monarch (Danaus plexippus) larval development on common milkweed, Asclepias syriaca. 3. In the glasshouse, water stress altered a suite of A. syriaca functional traits but did not affect nutrient content, whereas herbivore damage increased leaf nitrogen (N) and reduced the carbon:nitrogen (C:N) ratio. A bioassay experiment showed that monarch larval survival was lower on well‐watered plants that were previously damaged by monarch larva than on damaged and drought‐stressed plants. Bioassay larvae consumed less leaf tissue of previously damaged plants, whereas monarch larval mass was affected additively by water stress and previous damage, after correcting for the amount of leaf tissue consumed. 4. In a 2‐year field experiment, monarch larval performance was higher on previously damaged A. syriaca plants that received experimentally reduced rainfall, relative to plants receiving ambient rainfall. 5. Collectively, these results from glasshouse and field experiments suggest that insect performance was highest on previously damaged plants under water stress and highlight the additive and interactive roles of abiotic and biotic factors on herbivore performance.     

Infection of ‘Candidatus Phytoplasma ulmi’ reduces the protein content and alters the activity of detoxifying enzymes in Amplicephalus curtulus

It has been reported that insecticide‐detoxifying enzymes such as glutathione S‐transferases (GST) and esterases are affected by microbial infections in hemipteran insect vectors. The total protein content, and GST and α‐ and β‐esterase activities were quantified in ‘Candidatus Phytoplasma ulmi’‐infected and uninfected adults of Amplicephalus curtulus Linnavuori & DeLong (Hemiptera: Cicadellidae) at 25, 35, and 45 days after the acquisition access period (AAP) in the head‐thorax and abdomen sections. The total protein content was lower in phytoplasma‐infected leafhoppers 25, 35, and 45 days after the AAP. Thirty‐five days after the AAP, the GST and β‐esterase activities had increased (26 and 69%, respectively) compared to the control. However, 45 days after the AAP, the phytoplasma‐infected leafhoppers displayed lower GST (87%) and β‐esterase (253%) activities than the uninfected individuals. On the other hand, the α‐esterase activity proved to be unaffected by the phytoplasma infection. Forty‐five days after the AAP, females had a higher phytoplasma titer (46%) in their head‐thorax than in their abdomen sections, whereas males showed a higher titer in their abdomens (75%). In addition, the GST and β‐esterase activities in the abdomen were affected negatively by 96–98% as a result of the increasing ‘Ca. Phytoplasma ulmi’ titer. These results indicate that an infection of ‘Ca. Phytoplasma ulmi’ alters the metabolic activities of A. curtulus.     

Effect of viruses on agronomic and brewing characteristics of four hop (Humulus lupulus) cultivars in Australia

The effect of Hop latent virus (HpLV), Hop mosaic virus (HpMV), and Prunus necrotic ring spot virus [PNRSV (apple, A, and intermediate, I, serotypes)], on the survival of softwood cuttings, the vigour (height) of early season growth, cone yield, and the levels of brewing organic acids in mature plants, was assessed in four hop (Humulus lupulus) cultivars in Tasmania, Australia. Virus infections were associated with an increase in the mortality of softwood cuttings following propagation. In all cultivars, height of early growth was a poor indicator of the effect of viruses on cone yield and levels of brewing organic acids (alpha and beta acids). In cv. ‘Nugget’, infection by the virus combinations studied was not associated with reductions in cone yield, however plants infected by PNRSV‐I, in 2000, had 11% lower alpha acids and 7% lower beta acids. In ‘Opal’, infection by HpLV and HpMV were the most deleterious to cone yield, however the effect of HpMV was ameliorated when in combination with PNRSV‐I. Reductions in alpha and beta acid content were attributable only to mixed infections of HpLV + HpMV in combination with either serotype of PNRSV. In ‘Pride of Ringwood’, yield loss was mostly attributable to HpMV and to a lesser extent, HpLV. Some ameliorations in cone yield loss occurred in plants containing a mixed infection between HpMV and HpLV or either of the ilarvirus serotypes. Both of the ilarviruses and HpMV caused reductions in alpha acid content. In ‘Victoria’, cone yield loss was mostly attributable to combinations of viruses such as HpLV + PNRSV‐I and HpLV + HpMV. The deleterious effect of HpLV + HpMV was ameliorated by PNRSV‐A and to a lesser extent, PNRSV‐I. Infection by the virus combinations studied did not significantly affect alpha and beta acid levels in either year. Results suggested the effect of viruses and their combinations differed between cultivars and varied between seasons. This information, when combined with knowledge of the rates of virus re‐infection, can be used to recommend control strategies for the Australian hop industry.     

Biochemical and epigenetic changes in phytoplasma‐recovered periwinkle after indole‐3‐butyric acid treatment

Aim: To elucidate the possible mechanism of phytoplasma elimination from periwinkle shoots caused by indole‐3‐butyric acid (IBA) treatment. Methods and Results: It has been shown that a transfer of in vitro‐grown phytoplasma‐infected Catharanthus roseus (periwinkle) plantlets from medium supplemented with 6‐benzylaminopurine (BA) to one supplemented with IBA can induce remission of symptoms and even permanent elimination of ‘Candidatus Phytoplasma asteris’ reference strain HYDB. Endogenous auxin levels and general methylation levels in noninfected periwinkles, periwinkles infected with two ‘Candidatus Phytoplasma’ species and phytoplasma‐recovered periwinkles were measured and compared. After the transfer from cytokinin‐ to auxin‐containing media, healthy shoots maintained their phenotype, methylation levels and hormone concentrations. Phytoplasma infection caused a change in the endogenous indole‐3‐acetic acid to IBA ratio in periwinkle shoots infected with two ‘Candidatus Phytoplasma’ species, but general methylation was significantly changed only in shoots infected with ‘Ca. P. asteris’, which resulted in the only phytoplasma species eliminated from shoots after transfer to IBA‐containing medium. Both phytoplasma infection and treatment with plant growth regulators influenced callose deposition in phloem tissue, concentrations of photosynthetic pigments and soluble proteins, H₂O₂ levels and activities of catalase (CAT) and ascorbate peroxidase (APX). Conclusion: Lower level of host genome methylation in ‘Ca. P. asteris’‐infected periwinkles on medium supplemented with BA was significantly elevated after IBA treatment, while IBA treatment had no effect on cytosine methylation in periwinkles infected with ‘Candidatus Phytoplasma ulmi’ strain EY‐C. Significance and Impact of the Study: Hormone‐dependent recovery is a distinct phenomenon from natural recovery. As opposed to spontaneously recovered plants in which elevated peroxide levels and differential expression of peroxide‐related enzymes were observed, in hormone‐dependent recovery changes in global host genome, methylation coincide with the presence/absence of phytoplasma.     

Effects of ‘Candidatus Phytoplasma asteris’ on the Volatile Chemical Content and Composition of Grindelia robusta Nutt.

Grindelia robusta, a perennial herb, contains an essential oil that is used as an antitussive, sedative, and analgesic agent. During the spring of 2007, ‘Candidatus Phytoplasma asteris’‐related phytoplasmas were identified in plants showing virescence and phyllody symptoms. The qualitative and quantitative composition of the oil of healthy and infected plants was compared by gas chromatography/mass spectrometry. Samples from six symptomatic and five asymptomatic plants tested by nested PCR followed by RFLP analyses confirmed the presence of ‘Ca. P. asteris’ in all symptomatic samples. The oils from healthy and infected plants, obtained by steam distillation, contained 42 components; that of healthy plants contained a higher concentration of monoterpenes, especially limonene and bornyl acetate, which were nearly 50% higher.     

Effects of endophyte infection on drought stress tolerance of Lolium perenne accessions from the Mediterranean region

Perennial ryegrass (Lolium perenne) is a cool-season grass species that is often infected by the leaf-inhabiting endophyte Neotyphodium lolii. This particular endophyte is asexual and has the potential to impact host survival, growth and reproduction. The objective was to assess the potential costs or benefits of endophyte infection on drought stress tolerance of native perennial ryegrass accessions originally collected from Italy, Morocco, Tunisia, and Turkey. Sixty infected (E+) individuals from each accession were planted in a greenhouse. Half of these individuals were treated with a systemic fungicide to eliminate the endophyte (E−). For two drought periods water was withheld for 10-14 days and then allowed a one week recovery period following each. In some accessions under drought, E+ plants had more tillers, greater tiller lengths, total dry mass and green shoot mass than E− plants, suggesting a positive effect of endophyte infection on host growth. Total tiller length and the number of tillers showed significant population × treatment × infection interactions for 4 of 6 populations. This work is one of the few that documents the effects of endophyte infection for a common forage grass species from wild populations native to its distributional range. The results demonstrate that endophyte infection can help ameliorate abiotic stress such as drought and there may be a selective advantage for grasses from certain Mediterranean regions.