Attempts to eradicate graft-transmissible infections through somatic embryogenesis in <Emphasis Type="Italic">Citrus</Emphasis> ssp. and analysis of genetic stability of regenerated plants

Ability to detect Pseudocercospora macadamiae infection in macadamia husk at least four months before symptoms become visible will aid the development of disease control measures. This study examined the distinctness of P. macadamiae within the phylogenetic lineages of the genus Pseudocercospora. In addition, we developed two quantitative PCR (qPCR) assays, as rapid diagnostic tools, for early detection and quantification of P. macadamiae in planta. Phylogenetic analysis of concatenated sequences of four gene loci (large subunits, internal transcribed spacer (ITS), translation elongation factor 1-alpha (TEF-1α) and actin of 47 P. macadamiae isolates showed that P. macadamiae is a distinct species in the genus Pseudocercospora. P. macadamiae isolates were partitioned into subunits in the cluster but the grouping of the isolates was regardless of location. Nucleotide diversity (0.02) and the coefficient of genetic differentiation (0.07) were low in the P. macadamiae population. Two qPCR primer sets, based on ITS (PMI) and TEF-1α (PME) were designed that consistently amplified P. macadamiae in fungal cultures (Ct = 16.93 ± 0.11 and Ct = 21.20 ± 0.11, respectively) and in planta (Ct = 32.36 ± 0.28 and Ct = 38.07 ± 1.20, respectively). The PMI primers also detected species in the genus Pseudocercospora, while PME was more specific and robust for quantification of P. macadamiae. Both primer sets detected P. macadamiae in asymptomatic tissue samples and strongly differentiated various stages of disease progression, which revealed approximately 10-fold increase in fungal biomass between each consecutive stage of symptom development.     

Assessment of latent infection with <Emphasis Type="Italic">Verticillium longisporum</Emphasis> in field-grown oilseed rape by qPCR

Improvement of cultivar resistance is the key strategy to control the host-specialized pathogen Verticillium longisporum in oilseed rape (OSR). A special feature of this pathogen is its systemic, non-homogenous and delayed colonization of the plant xylem resulting in an extended symptomless period of latency. As a result, severity of infection in the field is difficult to score as it becomes apparent only at crop maturity stages when it may be confused with natural senescence. Assessment of Verticillium disease severity in OSR by visual scoring of microsclerotia on harvested stubbles unsatisfactorily reflects genotypic resistance as it is strongly affected by the ripening stage of the plant. To overcome these limitations, we developed a qPCR method, which unambiguously differentiates levels of quantitative resistance to V. longisporum in OSR genotypes under field conditions. The specificity and sensitivity of two primer pairs targeting ITS or tubulin loci in the V. longisporum genome were tested. While tubulin primers showed a high specificity to V. longisporum isolates, ITS primers exhibited a significantly higher sensitivity in detecting fungal DNA in stem tissue (limit of quantification =0.56 fg DNA) of field-grown pre-symptomatic plants. The best discrimination of resistant and susceptible OSR cultivars based on fungal DNA analysis in stem tissue was achieved at growth stage 80, at the transition of fungal vascular growth in viable plants to saprotrophic colonization of senescent stem tissues. Field screening data obtained with qPCR at growth stage 80 confirmed results from greenhouse testing thus corroborating the relevance and reliability of seedling assays for determining cultivar responses to V. longisporum in the field, as a useful tool for breeders in first selection of elite OSR genotypes with improved resistance to Verticillium.     

Interaction of root colonizing biocontrol agents demonstrates the antagonistic effect against <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">lycopersici</Emphasis> on tomato

Plant growth promoting Bacillus subtilis MSS9 and Bacillus licheniformis MSS14 were isolated from the tomato rhizosphere. These isolates were capable of inhibiting the fungal pathogen, Fusarium oxysporum f. sp. lycopersici causing fusarium wilt in tomato, tested by dual culture method and by mycolytic enzyme production. The isolates have the capacity to form biofilm on the microtitre plate. Scanning electron microscopy revealed good colonization capacity of Bacillus licheniformis MSS14 on tomato plant root as compared to Bacillus subtilis MSS9, pot experiments were also analyzed to study the effects of both rhizobacterial cultures on pathogen development and plant growth. It was observed that MSS14 reduces the incidence of Fusarium oxysporum f. sp. lycopersici in tomato and there was significant increase in vegetative parameters like root length, shoot length, plant wet weight, dry weight and chlorophyll content after which indicates that the root colonization property of the culture MSS14 helps in enhancing the biocontrol capacity against pathogen than that of MSS9.     

Local infection associated with vascular bundles: the colonization of coconut palm leaflets by two <Emphasis Type="Italic">Camarotella</Emphasis> species

Two Camarotella (Phyllachoraceae) species, C. torrendiella and C. acrocomiae are the causative agents of small (SV) and large verrucosis (LV), respectively, which are important diseases affecting Brazilian coconut palms. The small verrucosis produces necrotic lesions in coconut palm leaflets, whereas LV just produces chlorosis. Semi-thin sections of asymptomatic leaflets and of leaves presenting stromata in different development stages were compared through light microscopy in order to characterize the colonization process of these two coconut palm verrucosis agents. Camarotella torrendiella initially colonized the adaxial epidermal cells and the cells underlying the epidermis close to the vascular bundles. In latter colonization stages, the hyphae of C. torrendiella remained limited to the underlying necrotic tissue adjacent to the mature stromata, mostly in the intracellular spaces and in the collapsing cells of necrotized tissues around the vascular bundles. This species does not colonize intracellular intact fiber cells, xylem vessels or phloem sieve tube elements. In contrast, C. acrocomiae presented a typical biotrophic parasitism model such as that of some gramineous Phyllachora spp. High densities of C. acrocomiae hyphae were found inside intact sieve tube elements; however, with no evidence of cellular death. The extensive hyphal colonization by C. acrocomiae within sieve tube elements was also observed in tertiary and quaternary bundles, as well as in anastomosing vascular bundles. The dependence of both species on the colonization sites associated with vascular bundles indicates the need for additional studies about these intricate host-pathogen relationships. These studies could be important to define new strategies to control coconut palm verrucosis diseases.     

Rice response to simultaneous bacterial blight and drought stress during compatible and incompatible interactions

Plant response to one type of stress can be affected by simultaneous exposure to a second stress, for example when abiotic and biotic stresses occur together. Ten rice genotypes comprising those with bacterial blight (BB) resistance (R) genes, drought quantitative trait loci (QTLs) plus a BB R gene, and BB susceptible genotypes, were subjected to mild and moderate drought stress and plants were inoculated with two Xoo strains (PXO99 and PXO145) to simulate the challenges rice crops face under simultaneous stress of drought and BB. Plant height and dry shoot biomass were significantly reduced by drought stress treatments. The BB disease lesion lengths varied according to rice genotypes and PXO99 Xoo multiplication and spread in planta was higher compared to that of PXO145, which generally decreased under mild drought stress. Rice genotype IRBB7 (Xa7) showed less Xoo spread and a reduced Xoo multiplication under drought stress compared to the well-watered control with PXO145. In contrast, in genotypes with a different BB R gene and/or drought QTLs [IRBB4 (Xa4), IR87705–6-9-B (Xa4 + qDYT _2.2 ), IR87707–445-B-B-B (Xa4 + qDYT _2.2  + qDYT _4.1 ) and IR87707–446-B-B-B (Xa4 + qDYT _2.2  + qDYT _4.1 )], Xoo multiplication and spread in planta was higher with drought stress. This study has shown that drought stress affected rice response to the BB pathogen and the response varied according to the rice genotype. It is concluded that evaluating rice varieties under combined abiotic and biotic stresses will be the best strategy to determine biotic stress resistance durability under climate change.     

Resistance evaluation of differentials and commercial wheat cultivars to stripe rust (<Emphasis Type="Italic">Puccinia striiformis</Emphasis>) infection in hot spot regions of Canada

Stripe rust of wheat, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most important diseases of wheat in Canada. This study presents the results from resistance evaluation of Yr genes and western Canadian wheat cultivars from different milling classes, to natural infection in southern Alberta and British Columbia which are considered hot spots of stripe rust occurrence in Canada, due to proximity to Pacific Northwest of the United States where stripe rust epidemics are frequent. Genes Yr1, Yr5, Yr15, and YrSP were effective in all environments; Yr17 and Yr28, which were earlier reported ineffective to existing stripe rust races at the seedling stage in Canada, were effective at adult plant stages in most of the environments because of warmer climates in southerly locations, a favourable condition for expression of the genes. Yr17 is common in winter wheat cultivars and only reported spring wheat cultivar carrying it is CDC Stanley, which can serve as donor parent in breeding programs. Gene Yr24/26 was not very effective in western prairies although reported as effective in eastern prairies. Residual resistance from combination of defeated genes (Yr3, Yr7, Yr9, Yr27) in some supplementary differentials was observed. Most cultivars carry slow-rusting, pleiotropic adult-plant resistance gene Yr18 and some Yr29, which were effective in some locations. These genes failed to provide complete protection under high disease pressure. Seedling and adult plant resistance genes Yr5, Yr15, Yr17 and Yr18, Yr36, respectively could be good targets for resistance breeding. Stacking adult plant resistance genes with seedling resistance genes can provide durable resistance to stripe rust.     

Antagonistic effects of biofilm-forming bacterial strains co-inoculated with blood disease pathogenic strain, <Emphasis Type="Italic">Ralstonia syzygii</Emphasis> subspecies <Emphasis Type="Italic">celebesensis</Emphasis> in banana plants

A blood disease pathogenic strain, Ralstonia syzygii subspecies celebesensis was used to study the possible association of biofilm-forming bacteria with the development and severity of blood disease in banana plants. Therefore, the objective of this study was to determine the effects of mono-culture and co-culture inoculation of isolated biofilm-forming bacteria with the blood disease pathogen in banana pseudostems in glasshouse conditions. Putative biofilm-forming bacteria were isolated from an infected banana plant and were further identified using 16SrRNA sequencing. Four isolates, identified as Enterobacter hormaechei, Enterobacter cloacae, Kosakonia radicincitans and Klebsiella pneumoniae, were inoculated as a mono- and co-culture with R. syzygii subsp. celebesensis into 2 months old banana plants. The observation after the 8 weeks of post inoculation showed that plants which were co-inoculated with the pathogen and K. radicincitans, a biofilm-forming bacterium, were the most susceptible towards the infection. In contrast, plants under two treatments (which were co-inoculated with the pathogen and E. cloacae and the pathogen with E. hormaechei) were less susceptible towards the infection. This study revealed the antagonistic effects of two biofilm-forming strains which reduced the severity of infection caused by the pathogenic agent. Scanning electron micrographs of the cross section of plant rhizomes indicated the dissimilarity of adhesion and host colonization conditions of the pathogen in each infected plant from different treatments.     

In vitro competition between <Emphasis Type="Italic">Fusarium graminearum</Emphasis> and <Emphasis Type="Italic">Epicoccum nigrum</Emphasis> on media and wheat grains

Competitive effects between Fusarium graminearum, causing Fusarium head blight, and the endophyte Epicoccum nigrum, were performed in in vitro competition assays between the two species. Two E. nigrum isolates were isolated from wheat grains and tested as competitors against two F. graminearum isolates. A dual petri dish assay showed that E. nigrum reduced the mycelial growth of F. graminearum and vice versa. A glass slide assay revealed that E. nigrum crude cultural filtrate also had reducing effect on the growth of F. graminearum comparable to that of E. nigrum spore suspensions. Microscopy showed hyphae of F. graminearum and E. nigrum with many side branches when in close proximity, in contrast to pronounced apical hyphal growth when growing alone. Combinations of F. graminearum and E. nigrum on sterilised wheat grains were studied over time by qPCR. F. graminearum biomass was significantly reduced in inoculations applying E. nigrum three days prior to F. graminearum. In conclusion, these results showed competition and mycelial behaviour effects between F. graminearum and E. nigrum and support that E. nigrum may have potential to reduce F. graminearum infections in wheat. Competition experiments should be carried out in planta to study the interaction further.     

Rice <Emphasis Type="Italic">OsPBL1</Emphasis> (ORYZA SATIVA ARABIDOPSIS PBS1-LIKE 1) enhanced defense of <Emphasis Type="Italic">Arabidopsis</Emphasis> against <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> DC3000

The receptor-like cytoplasmic kinases (RLCK family VII) are required for plant defense against various pathogens. Previously, OsPBL1 (ORYZA SATIVA ARABIDOPSIS PBS1-LIKE 1) was isolated from rice as a potential RSV (rice stripe virus) resistant factor, but its physiological roles in plant defense are yet to be investigated. In this study, we demonstrated that OsPBL1increased defense against P. syringae in transgenic Arabidopsis. To ascertain the role of OsPBL1 gene in plant defense, OsPBL1 tagged with HA (i.e. Hemagglutinin) was overexpressed in Arabidopsis and examined for the resistance against Pseudomonas syringae pv. tomato DC3000 (i.e. Pst DC3000). At 3 dpi of Pst DC3000, transgenic Arabidopsis lines exhibited the reduced chlorotic lesion and propagation of P. syringae, compared to wild type. Elevated pathogen resistance of transgenic lines was correlated with increased H₂O₂ accumulation and callose deposition on the infected leaves. It was also revealed that expression levels of salicylic acid dependent genes such as PR1, PR2, and PR5, were induced higher in transgenic lines than wild type. Taken together, our data suggested that OsPBL1 exerted the role in defense against pathogen attacks in plant via mainly facilitating salicylic acid dependent pathway.     

AlgU contributes to the virulence of <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">tomato</Emphasis> DC3000 by regulating production of the phytotoxin coronatine

Pseudomonas syringae pv. tomato DC3000 (Pst DC3000), which causes bacterial speck disease of tomato, has been used as a model pathogen to investigate the molecular basis of plant–pathogen interactions. The function of many potential virulence factors encoded in the Pst DC3000 genome and their modes of action are not fully understood. P. syringae is known to produce the exopolysaccharide alginate. Although AlgU, a sigma factor, is known to regulate the expression of genes such as algD related to alginate biosynthesis, the molecular mechanisms of AlgU in the virulence of Pst DC3000 is still unclear. To investigate the function of AlgU and alginate in plant–bacterial pathogen interactions, we generated ΔalgU and ΔalgD mutants. After inoculation with ΔalgU but not ΔalgD, host plants of Pst DC3000 including tomato and Arabidopsis had milder disease symptoms and reduced bacterial populations. Expression profiles of Pst DC3000 genes revealed that AlgU can regulate not only the expression of genes encoding alginate biosynthesis, but also the expression of genes related to type III effectors and the phytotoxin coronatine (COR). We also demonstrated that the ΔalgU mutant showed full virulence in the Arabidopsis fls2 efr1 double mutant, which is compromised in the recognition of PAMPs. Further, the application of COR was able to restore the phenotype of the ΔalgU mutant in the stomatal response. These results suggest that AlgU has an important role in the virulence of Pst DC3000 by regulating COR production.     

Effect of simultaneous and sequential inoculations of <Emphasis Type="Italic">Meloidogyne incognita</Emphasis>, <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> and <Emphasis Type="Italic">Phomopsis vexans</Emphasis> on eggplant in sand mix and fly ash mix soil

Effects simultaneous and sequential inoculations of Meloidogyne incognita, Ralstonia solanacearum and Phomopsis vexans were studied on the growth, chlorophyll and carotenoid contents of eggplants grown in 25% fly ash and 25% sand mix soil. Plants grown in 25% fly ash mix soil had lesser plant growth than grown in 25% sand ash mix soil. Inoculation of M. incognita / R. solanacearum or P. vexans caused reduction in plant growth, chlorophyll and carotenoid contents in both types of soils but these pathogens in combination caused a greater reduction in than individual inoculation. Inoculation of M. incognita 20 days prior to R. solanacearum caused a greater reduction in plant growth than inoculation of M. incognita prior to P. vexans. Inoculation of P. vexans prior to R. solanacearum caused a lesser reduction in plant growth, chlorophyll and carotenoid contents than inoculation of P. vexans prior to M. incognita. Inoculation of R. solanacearum 20 days prior to M. incognita caused a greater reduction in plant growth, chlorophyll and carotenoid contents than inoculation of R. solanacearum prior to P. vexans. Galling and multiplication of M. incognita was higher in plants grown in 25% sand amended soil than with 25% fly ash soil. R. solanacearum and P. vexans had adverse effects on galling and nematode multiplication. Wilt and blight indices caused by R. solanacearum and P. vexans were 3 respectively. Wilt and blight indices were 4 when two pathogens were inoculated together.     

Quantitative phenotyping of grey leaf spot disease in maize using real-time PCR

Grey leaf spot is an important maize foliar disease caused by the fungal pathogens Cercospora zeae-maydis and Cercospora zeina. Although methods exist to detect these Cercospora species in maize, current techniques do not allow quantification of the fungi in planta. We developed a real-time SYBR® Green PCR assay for quantification of grey leaf spot disease in maize based on the amplification of a fragment of a cytochrome P450 reductase (cpr1) gene. In planta fungal DNA content was normalised to a maize glutathione S-transferase III gene (gst3) to yield values of ng Cercospora DNA/mg maize DNA. The assay was specific to the two Cercospora spp., and we observed no amplification of the cpr1 fragment in non-target maize leaf pathogens or saprophytes. The assay was employed to quantify C. zeina in glasshouse inoculated maize plants and grey leaf spot infected field plants of resistant and susceptible maize lines. In both instances, C. zeina DNA content correlated with symptomatic leaf lesion area, and the susceptible maize line contained significantly more C. zeina DNA than the resistant line. Sequence differences between the C. zeina and C. zeae-maydis cpr1 amplicons enabled us to perform melt curve analyses to identify the Cercospora species causing grey leaf spot at a particular location. This assay has application in the early detection and quantification of Cercospora spp., both of which are important tools in grey leaf spot disease management and maize breeding programmes.     

Survival and development of spotted bollworm, <Emphasis Type="Italic">Earias vittella</Emphasis> (Fabricius) (Lepidoptera: Nolidae) on different transgenic <Emphasis Type="Italic">Bt</Emphasis> and isogenic non-<Emphasis Type="Italic">Bt</Emphasis> cotton genotypes

Four Bt cotton hybrids, each with one of four different events, viz., MRC 6301 Bt (cry1Ac gene), JKCH 1947 Bt (modified cry1Ac gene), NCEH 6R Bt (fusion cry1Ac/cry1Ab gene) and MRC 7017 Bollgard II (cry1Ac and cry2Ab genes) were compared for survival and development of Earias vittella (Fabricius) along with their isogenic non-Bt genotypes. None of the neonates were able to complete the larval period and reach pupal stage on squares of 90, 120 and 150 days old crop of all Bt hybrids. Likewise, on bolls also, zero per cent larval survival was observed in all Bt hybrids except JKCH 1947 Bt where 0.67 per cent larvae could manage to reach pre-pupal stage at 120 and 150 days old crop but failed to form cocoon and enter pupal stage. The surviving larva took more development time (3.7 to 5.4 days) as compared to larvae fed on bolls of JKCH 1947 non-Bt. The average survival period (ASP) of larvae was in order of 150 > 120 > 90 days old crop among the crop ages; JKCH 1947 Bt > MRC 6301 Bt > NCEH 6 R Bt > MRC 7017 Bollgard II among Bt hybrids; and bolls > squares between fruiting bodies. However, reverse was true for speed index of toxic effect. The concentration of Cry toxin varied significantly in squares and bolls and also among the crop ages. The amount of Cry toxin in squares and bolls had significant negative correlation with ASP of the E. vittella larvae.     

Chitin elicitor receptor kinase 1 (CERK1) is required for the non-host defense response of <Emphasis Type="Italic">Arabidopsis</Emphasis> to <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. Sp. <Emphasis Type="Italic">cubense</Emphasis>

Banana wilt disease is a typical vascular disease caused by the fungal pathogen Fusarium oxysporum f. sp. cubense 4 (Foc 4). Pattern recognition receptors in the plant cell membrane can recognize pathogen-associated molecular patterns (PAMPs) to activate multi-layer defense responses, including defense gene expression, stomatal closure, reactive oxygen species (ROS) burst and callose deposition, to limit pathogen growth. In the present study, we found that chitin elicitor receptor kinase 1 (CERK1) was required for the non-host resistance of Arabidopsis thaliana to Foc B2 (a strain of Foc 4). The cerk1 mutant had weaker defense responses after Foc B2 treatment, including lower expression of PAMP- and salicylic acid-responsive genes, no stomatal closure, lower ROS level and less callose deposition, than that of the wild-type plant. Consistent with this, the cerk1 mutant plants exhibited higher susceptibility to non-host pathogen Foc B2. These results suggest the crucial importance of CERK1 in Foc B2-triggered non-host resistance.     

Diversity of the cultivable gut bacterial communities associated with the fruit flies <Emphasis Type="Italic">Bactrocera dorsalis</Emphasis> and <Emphasis Type="Italic">Bactrocera cucurbitae</Emphasis> (Diptera: Tephritidae)

Gut microbes play an important role in insect morphogenesis, nutrition, development of resistance against parasitoids and detoxification of toxic compounds. A culture-based approach is therefore an useful tool for the characterization of cultivable microbial communities associated with the insect gut. In the present study an attempt was made to decipher the gender specificity of gut bacterial communities of two major fruit fly species of India viz., Bactrocera dorsalis (Hendel) and Bactrocera cucurbitae (Conquillett) (Diptera: Tephritidae). Based on molecular identification, B. dorsalis females were found to predominantly harbor the bacterial species Enterobacter cloacae, Enterobacter asburiae and Citrobacter freundii, while B. dorsalis males were found to harbor Providencia rettgerii, Klebsiella oxytoca, Enterococcus faecalis and Pseudomonas aeruginosa The cultivable diversity from females of B. cucurbitae comprised mainly of Morganella morganii and Bacillus pumilis while B.cucurbitae males were predominantly colonized by aerobic endospore formers viz., Bacillus cereus, B. licheniformis and B. subtilis. The above findings have thrown light on a distinct pattern of gender specific gut bacterial colonization in fruit flies, which have to be factored in for the formulation of fruit fly management strategies.     

The Eurotiomycete <Emphasis Type="Italic">Apinisia graminicola</Emphasis> as the causal agent of a leaf spot disease on the energy crop <Emphasis Type="Italic">Miscanthus</Emphasis> x <Emphasis Type="Italic">giganteus</Emphasis> in Northern Germany

Miscanthus x giganteus is a fast growing, perennial energy crop for temperate climates. Because of its high annual biomass production rates and its characteristics as a low-input crop, an expansion of field cultivation can be anticipated to cover increasing demands for sustainable biomass production. However, knowledge about pathogens that could have an impact on biomass production is still limited for M. giganteus. Here, we report about the isolation of the filamentous fungus Apinisia graminicola from necrotic leaf lesions of M. giganteus grown on a field trial plot in Northern Germany. Inoculation assays with the isolated A. graminicola strain confirmed its capacity to cause a leaf spot disease on M. giganteus. Additional inoculation assays revealed that A. graminicola also caused necrotic lesions on leaves of the model grass Brachypodium distachyon. Generally, symptoms of A. graminicola-caused leaf spot disease were stronger on B. distachyon compared to M. giganteus. Incubation temperatures above 22 °C during A. graminicola infection resulted in stronger disease symptoms on both, M. giganteus and B. distachyon leaves. Microscopic analysis of cross sectioned, infected leaf tissue revealed an epiphytic mycelium formation on the surface and an endophytic colonization of the mesophyll leave tissue, especially in M. giganteus. Our results revealed that the isolated A. graminicola strain is a causal agent of a leaf spot disease on grass leaves. Its potential on endophytic growth in M. giganteus might open new possibilities in studying this type of plant-fungal interaction on a cellular and molecular level in an energy crop.     

A chalcone synthase controls the verticillium disease resistance response in both <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> and cotton

Verticillium wilt is a devastating disease caused by the soil-borne fungus Verticillium dahliae that causes severe wilt symptoms in more than 400 plant species, including economically important cotton. However, the molecular mechanism of plant resistance to Verticillium remains unclear. In this study, we identified an Arabidopsis mutant, vsad1 (verticillium sensitive and anthocyanin deficient 1), which showed more serious disease symptoms such as discoloration and chlorosis than wild-type Arabidopsis. vsad1 is a previously identified allele of the transparent testa 4 gene (tt4), which encodes chalcone synthase (CHS), a key enzyme involved in the biosynthesis of flavonoids. Our results showed that VSAD1 expression was induced in response to Verticillium dahliae infection. Overexpression of VSAD1 partially recovered the anthocyanin accumulation phenotype of the vsad1–1 mutant. The concentration of V. dahliae increased and ROS accumulation decreased in the vsad1 mutant after infection with V. dahliae. Knockdown of the homologous gene GhCHS in cotton plants increased their susceptibility to V. dahliae infection. Thus, we conclude that VSAD1 is involved in the regulation of plant resistance to Verticillium wilt.     

Five plant families support natural sporulation of <Emphasis Type="Italic">Cronartium ribicola</Emphasis> and <Emphasis Type="Italic">C. flaccidum</Emphasis> in Finland

Fusarium is one of the most destructive fungal genera whose members cause many diseases on plants, animals, and humans. Moreover, many Fusarium species secrete mycotoxins (e.g. trichothecenes and fumonisins) that are toxic to humans and animals. Fusarium isolates from date palm trees showing disease symptoms, e.g. chlorosis, necrosis and whitening, were collected from seven regions across Saudi Arabia. After single-sporing, the fungal strains were morphologically characterized. To confirm the identity of morphologically characterized Fusarium strains, three nuclear loci, two partial genes of translation elongation factor 1 α (tef1α) and β-tubulin (tub2), and the rDNA-ITS region, were amplified and sequenced. Of the 70 Fusarium strains, 70 % were identified as F. proliferatum that were recovered from six regions across Saudi Arabia. Fusarium solani (13 %), as well as one strain each of the following species: F. brachygibbosum, F. oxysporum, and F. verticillioides were also recovered. In addition, five Fusarium-like strains were recognized as Sarocladium kiliense by DNA-based data. The preliminary in vitro pathogenicity results showed that F. proliferatum had the highest colonization abilities on date palm leaflets, followed by F. solani. Although F. oxysporum f. sp. albedinis is the most serious date palm pathogen, F. proliferatum and F. solani are becoming serious pathogens and efforts should be made to restrict and control them. In addition, the potential toxin risks of strains belonging to F. proliferatum should be evaluated.