Characterization of Fusarium diseases on commercially grown orchids in Hawaii

A decline of unknown aetiology has become a major problem for commercial orchid production in Hawaii, one of the primary orchid‐producing states in the USA. The major symptoms of decline include root degradation, foliar blight, pseudobulb rot and sheath rot. It was unclear whether all these symptoms are caused by the same or different pathogens, but preliminary research indicated that Fusarium species may be involved. In this study, the incidence of Fusarium species was examined across 186 plants, from 29 orchid genera and intergeneric hybrids across three islands in the state of Hawaii. The main five species associated with diseased orchids were F. proliferatum (38% of samples), F. solani (16%), F. oxysporum (16%) and two previously undescribed species (8% for both species combined). The two undescribed species were similar in appearance to F. subglutinans, and were designated FS‐A and FS‐B. Pathogenicity tests established that both F. proliferatum and FS‐B caused foliar spots, foliar blight and pseudostem rot on Dendrobium orchids, and that F. proliferatum isolates from diseased tissue of several genera could also induce symptoms on Dendrobium orchids. Although orchids have increasing importance in floriculture, relatively little is known about orchid pathogens, and previous studies focused primarily on Cymbidium and Phalaenopsis. This study provides new information concerning Dendrobium orchid pathogens and suggests a much wider host range than previously recognized for the five Fusarium species recovered from tissue with symptoms. These findings can contribute to better management of Fusarium diseases, which represent a significant challenge to orchid production in Hawaii.     

Stem rot of jewel orchids caused by a new forma specialis,Fusarium oxysporum f. sp. anoectochili in Taiwan

Stem rot of Anoectochilus formosanus (Af) caused by Fusarium oxysporum (Fo) is a major limiting factor to jewel orchid production in Taiwan. Fo causes discoloration in vascular tissues. However, some newly collected Fo isolates from Af stem rot do not cause vascular discoloration, suggesting changes may have occurred in the pathogen. Among recent Fo isolates from Af there are two colony types, the cottony alba (CA) and the sporodochial (S). In order to confirm that both colony types cause Af stem rot, 200 isolates were obtained from diseased stems in Nantou County and characterized by colony type and whether or not the infected plants had vascular discoloration. Isolates of both the CA and S types caused stem rot of Af; some isolates in each colony type caused vascular discoloration whilst others did not. Pathogenicity tests with 22 isolates resulted in stem rot disease severity ratings on Af of 3·1–4·0 and 2·1–4·0 with CA and S type colonies, respectively. The same isolates failed to cause disease on Cattleya, Dendrobium or Phalaenopsis plants. Phylogenetic analysis of partial intergenic spacer sequences showed that these isolates were distinguishable from other formae speciales of Fo and could be separated into two groups correlated with the CA or S type colonies with high bootstrap. Based on pathogenic, morphological and molecular characterizations, the Fo that causes stem rot of Af is proposed to be a new forma specialis, F. oxysporum f. sp. anoectochili, with different pathotypes.     

Genetic diversity of Fusarium oxysporum and related species pathogenic on tomato in Algeria and other Mediterranean countries

In order to characterize the pathogen(s) responsible for the outbreak of fusarium diseases in Algeria, 48 Fusarium spp. isolates were collected from diseased tomato in Algeria and compared with 58 isolates of Fusarium oxysporum originating from seven other Mediterranean countries and 24 reference strains. Partial sequences of the translation elongation factor EF‐1α gene enabled identification of 27 isolates as F. oxysporum, 18 as F. commune and three as F. redolens among the Algerian isolates. Pathogenicity tests confirmed that all isolates were pathogenic on tomato, with disease incidence greater at 28°C than at 24°C. All isolates were characterized using intergenic spacer (IGS) DNA typing, vegetative compatibility group (VCG) and PCR detection of the SIX1 (secreted in xylem 1) gene specific to F. oxysporum f. sp. lycopersici (FOL). No DNA polymorphisms were detected in the isolates of F. redolens or F. commune. In contrast, the 27 Algerian isolates of F. oxysporum were shown to comprise nine IGS types and 13 VCGs, including several potentially new VCGs. As none of the isolates was scored as SIX1+, the 27 isolates could be assigned to F. oxysporum f. sp. radicis‐lycopersici (FORL). Isolates from Tunisia were also highly diverse but genetically distinct from the Algerian isolates. Several Tunisian isolates were identified as FOL by a PCR that detected the presence of SIX1. The results show that isolates from European countries were less diverse than those from Tunisia. Given the difference between Algerian populations and populations in other Mediterranean countries, newly emergent pathogenic forms could have evolved from local non‐pathogenic populations in Algeria.     

Genetic variability among isolates of Fusarium oxysporum from sugar beet

Fusarium yellows, caused by the soil‐borne fungus Fusarium oxysporum f. sp. betae (Fob), can lead to significant yield losses in sugar beet. This fungus is variable in pathogenicity, morphology, host range and symptom production, and is not a well characterized pathogen on sugar beet. From 1998 to 2003, 86 isolates of F. oxysporum and 20 other Fusarium species from sugar beet, along with four F. oxysporum isolates from dry bean and five from spinach, were obtained from diseased plants and characterized for pathogenicity to sugar beet. A group of sugar beet Fusarium isolates from different geographic areas (including nonpathogenic and pathogenic F. oxysporum, F. solani, F. proliferatum and F. avenaceum), F. oxysporum from dry bean and spinach, and Fusarium DNA from Europe were chosen for phylogenetic analysis. Sequence data from β‐ tubulin, EF1α and ITS DNA were used to examine whether Fusarium diversity is related to geographic origin and pathogenicity. Parsimony and Bayesian MCMC analyses of individual and combined datasets revealed no clades based on geographic origin and a single clade consisting exclusively of pathogens. The presence of FOB and nonpathogenic isolates in clades predominately made up of Fusarium species from sugar beet and other hosts indicates that F. oxysporum f. sp. betae is not monophyletic.     

晋南地区黄瓜、黑籽南瓜死秧的病原菌鉴定及品种抗性研究

经分离、培养对不同菌种培养性状的观察,确定了侵染黄瓜、黑籽南瓜造成死秧的镰刀菌主要为尖镰孢菌黄瓜专化型、尖镰孢菌西瓜专化型、串珠镰刀菌和腐皮镰孢菌4种。经致病性测定,4种镰刀菌均能侵染黄瓜,引起发病造成死秧,可分为强致病类型和中强致病类型。经抗病性鉴定,黑籽南瓜种子只有南瓜4号为耐病品种;黄瓜种子也只有津优31号为耐病品种。     

Morphology,phylogeny and pathogenicity of Fusarium species from Sansevieria trifasciata in Malaysia

Leaf blight is a common disease affecting Sansevieria trifasciata in many countries, including Malaysia. In the present study, Fusarium isolates were consistently recovered from the diseased leaves collected from various locations throughout the country. Based on morphology and multigene phylogenetic analysis using mitochondrial small subunit (mtSSU), intergenic spacer region (IGS) and translation elongation factor 1-α (TEF1-α) gene sequences, seven Fusarium species were identified, with F. oxysporum being the most prevalent (67.6%) among 34 isolates. Pathogenicity tests resulted in the discovery of pathogenic isolates that belonged to F. oxysporum, F. proliferatum, and F. pseudocircinatum, whereas all isolates of F. brachygibbosum, F. concentricum, F. mangiferae, and F. solani were nonpathogenic. The results suggest that several Fusarium species are accountable for causing disease on S. trifasciata in Malaysia.     

Genetic diversity and pathogenicity of <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> isolated from wilted rocket plants in Italy

Thirty-six isolates of Fusarium oxysporum originated from Eruca vesicaria and Diplotaxis tenuifolia together with eight reference strains belonging to the formae speciales raphani, matthioli and conglutinans, typical on the Brassicaceae family, were tested for pathogenicity on two species of rocket plants (E. vesicaria L., syn. E. sativa, cv. ‘Rucola coltivata’; and D. tenuifolia cv. ‘Winter’) cultivated in the glasshouse. The results showed that different isolates were slightly, moderately or highly virulent. The strains were examined for differences in the nucleotide sequence of the ribosomal DNA (rDNA) intergenic spacer (IGS) region, about 2.5 kb long. The phylogenetic (neighbor-joining) analysis performed on the isolates enabled identification of four different groups, named I, II, III and IV. Thirty-one isolates out of 36 clustered in group I and were genetically similar to F. oxysporum f.sp. raphani. By considering the pathogenicity of the strains included in Group I, a partial host specialization could be observed: the average disease index of the isolates from D. tenuifolia was higher on wild rocket, whereas the average disease index of the isolates from E. vesicaria was higher on cultivated rocket. Moreover, isolates from cultivated rocket showed, on average, a higher degree of aggressiveness than the isolates from wild rocket. Concerning Group I, the sequence analysis confirmed the homogeneity of the population, with only five parsimony-informative SNPs and five haplotypes. Twenty-six out of 31 isolates belonged to haplotype 1. Groups II and III were genetically similar to strains of F. oxysporum f.sp. matthioli. Three other strains, not pathogenic or with a medium level of virulence, clustered together in Group 4, but their sequence was distant from that of other formae speciales. The pathogenicity and IGS analysis confirmed the presence of virulence variation and genetic diversity among the F. oxysporum isolates studied. To our knowledge, this is the first report of differentiation of formae speciales of F. oxysporum on rocket plants by IGS analysis.     

Isolation and characterization of novel soilborne lytic bacteriophages infecting Dickeya spp. biovar 3 (‘D. solani’)

Nine bacteriophages infecting Dickeya spp. biovar 3 (‘Dickeya solani’) were isolated from soil samples collected in different regions in Poland. The phages have a typical morphology of the members of the order Caudovirales, family Myoviridae, with a head diameter of c. 90–100 nm and tail length of c. 120–140 nm. In host range experiments, phage ?D5 expressed the broadest host range, infecting members of all Dickeya spp., and phage ?D7 showed the narrowest host range, infecting isolates of Dickeya dadantii and ‘D. solani’ only. None of the phages was able to infect Pectobacterium spp. isolates. All phages were prone to inactivation by pH 2, temperature of 85°C and by UV illumination for 10 min (50 mJ cm^?2). Additionally, phages ?D1, ?D10 and ?D11 were inactivated by 5 m NaCl and phage ?D2 was inactivated by chloroform. Phages ?D1, ?D5, ?D7 and ?D10 were characterized for optimal multiplicity of infection and the rate of adsorption to the bacterial cells. The latent period was 30 min for ?D1, 40 min for ?D5, 20–30 min for ?D7 and 40 min for ?D10. The estimated burst size was c. 100 plaque‐forming units per infected cell. The bacteriophages were able to completely stop the growth of ‘D. solani’ in vitro and to protect potato tuber tissue from maceration caused by the bacteria. The potential use of bacteriophages for the biocontrol of biovar 3 Dickeya spp. in potato is discussed.     

Identification and distribution of fungal pathogens associated with seedling blight of rice in the southern United States

Surveys were conducted in the five southern rice-producing states of Arkansas, Louisiana, Mississippi, Missouri and Texas in the United States during the 2018 and 2019 cropping seasons to determine the distribution and pathogenicity of fungal pathogens associated with seedling blight in rice. A total of 349 pathogenic fungal isolates were collected and identified as belonging to four genera: Rhizoctonia solani, Fusarium spp., Sclerotium rolfsii and Marasmius graminum based on morphological characteristics, molecular analysis and Koch's postulates. R. solani (252 out of 349 pathogenic isolates) was the most prevalent fungus isolated from diseased samples. Of the 252 pathogenic R. solani isolates, 245 were further classified as anastomosis group 11 (AG-11) and 7 as AG-4. Isolates of R. solani AG-4 and M. graminum were the most aggressive, with the highest stand loss (63% to 100%) and median disease rating (DR; 5.0), followed by isolates of R. solani AG-11 (stand loss = 4% to 100% and DR = 0.6 to 5.0), Fusarium spp. (stand loss = 26% to 48% and DR = 2.0 to 5.0) and S. rolfsii (stand loss = 33% to 48% and DR = 2.0 to 3.0) in causing seedling blight in rice. R. solani (62% to 83% of total pathogenic isolates) and Fusarium spp. (10% to 24% of total pathogenic isolates) were predominant in all the five states surveyed. S. rolfsii and M. graminum were present only in Louisiana and Texas. The results of this first systematic survey of rice seedling diseases in the southern United States will help develop effective fungicide seed treatment strategies for control of stand loss caused by seedling blight, one of the major factors limiting rice production.     

Phylogenetic relationships and virulence assays of Fusarium secorum from sugar beet suggest a new look at species designations

Fusarium spp. are responsible for significant yield losses in sugar beet (Beta vulgaris) with Fusarium oxysporum f. sp. betae most often reported as the primary causal agent. Recently, a new species, F. secorum, was reported to cause disease in sugar beet but little is known on the range of virulence within F. secorum or how this compares to the virulence and phylogenetic relationships previously reported for Fusarium pathogens of sugar beet. To initiate this study, partial translation elongation factor 1-α (TEF1) sequences from seven isolates of F. secorum were obtained and the data were added to a previously published phylogenetic tree that includes F. oxysporum f. sp. betae. Unexpectedly, the F. secorum strains nested into a distinct group that included isolates previously reported as F. oxysporum f. sp. betae. These results prompted an expanded phylogenetic analysis of TEF1 sequences from genomes of publicly available Fusarium spp., resulting in the additional discovery that some isolates previously reported as F. oxysporum f. sp. betae are F. commune, a species that is not known to be a sugar beet pathogen. Inoculation of sugar beet with differing genetic backgrounds demonstrated that all Fusarium strains have a significant range in virulence depending on cultivar. Taken together, the data suggest that F. secorum is more widespread than previously thought. Consequently, future screening for disease resistance should rely on isolates representing the full diversity of the Fusarium population that impacts sugar beet.     

Identification,pathogenicity and community dynamics of fungi and oomycetes associated with pea root rot in northern France

The pea root rot complex is a major concern for green pea production worldwide. This study aimed at characterizing its composition and dynamics throughout a cropping season in northern France. To this end, fungi and oomycetes were isolated from green pea plant roots with symptoms sampled at the flowering stage in 22 fields in 2017, and at the pea emergence, elongation and flowering stages in two fields in 2018. Out of 646 isolates collected, 317 were identified using molecular markers. Fusarium oxysporum, F. solani and F. redolens were highly predominant. Pathogenicity tests separated the isolates into four aggressiveness groups. F. solani isolates were the most aggressive. Phylogenetic analysis of their TEF1 sequences showed that they mainly belonged to the F. pisi lineage, and that F. oxysporum isolates were genetically close to isolates from the UK that did not belong to the forma specialis pisi. In addition, several Clonostachys rhizophaga isolates are reported for the first time to cause pea root rot. The oomycetes were rarely found and were represented by a few Pythium spp. isolates. Lastly, this study shows that the fungal and oomycete communities associated with pea root rot change during the cropping season. The level of dissimilarity of the root-rot-associated communities decreased throughout the cropping season towards a more similar composition at the flowering stage, dominated by F. solani, F. oxysporum and F. redolens. The proportion of nonpathogenic to weakly pathogenic isolates decreased progressively during the growing season in favour of moderately to highly pathogenic isolates.     

Phylogenetic relationships of Fusarium oxysporum f. sp. melonis in Iran

Fusarium wilt of melon caused by Fusarium oxysporum f. sp. melonis is a destructive fungal disease in melon growing regions. Isolates of F. oxysporum obtained from six major melon producing provinces in Iran, from melons and other hosts, were characterized based on pathogenicity to melon, vegetative compatibility groups (VCGs) and nuclear ribosomal DNA intergenic spacer (IGS) sequencing. Thirty-four of 41 isolates from Iran in this study were identified as race 1,2 which belonged to either VCG 0134 or an unassigned VCG, which based on IGS sequencing grouped with the VCG 0135 tester isolate. The seven remaining isolates were identified as nonpathogenic to melon belonging to two undescribed VCGs. Based on sequence analyses of the IGS region of Iranian and foreign isolates, nine lineages were identified, each including one VCG. The separation of VCGs into distinct lineages based on IGS sequences is mostly consistent with Repetitive extragenic palindromic PCR (Rep-PCR) results. Exceptions are VCGs 0130 and 0131, which could be differentiated with IGS sequences, but not with Rep-PCR. Different races from the USA, France and Iran associated with VCG 0134 grouped into one IGS lineage but could be differentiated with Rep-PCR, suggesting that this VCG is more diverse than previously thought. Given the long history of melon cultivation in Iran and the Rep-PCR diversity of isolates belonging to this VCG, it could be speculated that VCG 0134 perhaps evolved in Iran.     

Diverse <Emphasis Type="Italic">Fusarium solani</Emphasis> isolates colonise agricultural environments in Ethiopia

Fusarium solani is a fungal pathogen that infects many different genera of plants. It represents one of the two Fusarium spp. commonly isolated from agricultural soils and plant tissues in Ethiopia. To determine the diversity of F. solani in Ethiopia, we studied 43 isolates using Amplified Fragment Length Polymorphism (AFLP) and nucleotide sequences of the Translation Elongation Factor 1α (TEF-1α) and β-tubulin genes. TEF-1α sequences from GenBank, representing previously described species and clades of the F. solani-Haematonectria haematococca complex, were also included for comparative purposes. Phylogenetic analyses of the TEF-1α data separated the isolates into three groups corresponding with the three previously described clades (Clades 1–3) for this fungus. The Ethiopian isolates aggregated into one group corresponding to Clade 3. TEF-1α, β-tubulin and AFLPs further separated the Ethiopian isolates into a number of clusters and apparently novel phylogenetic lineages. Although the biological and ecological significance of these lineages and clusters is unclear, our data show that the Ethiopian agricultural environment is rich in species and lineages of the F. solani-H. haematococca complex.     

Occurrence of the F129L mutation in Alternaria solani populations in Germany in response to QoI application,and its effect on sensitivity

Early blight caused by Alternaria solani is a highly destructive disease of potatoes. Control of early blight mainly relies on the use of preventive fungicide treatments. Because of their high efficacy, azoxystrobin and other quinone outside inhibitors (QoIs) are commonly used to manage early blight. However, loss of sensitivity to QoIs has previously been reported for A. solani in the United States. Two hundred and three A. solani field isolates collected from 81 locations in Germany between 2005 and 2011 were screened for the presence of the F129L mutation in the cytochrome b gene; of these, 74 contained the F129L mutation. Sequence analysis revealed the occurrence of two structurally different cytb genes, which differed in the presence (genotype I) or absence (genotype II) of an intron, with genotype I being the most prevalent (63% of isolates). The F129L mutation was detected only in genotype II isolates, where it occurred in 97%. Sensitivity to azoxystrobin and pyraclostrobin was determined in conidial germination assays. All isolates possessing the F129L mutation had reduced sensitivity to azoxystrobin and, to a lesser extent, to pyraclostrobin. Early blight disease severity on plants treated with azoxystrobin was significantly higher for A. solani isolates with reduced fungicide sensitivity in the conidial germination assay compared with sensitive isolates. Data suggest an accumulation of F129L isolates in the German A. solani population over the years 2009–2011. It is assumed that the application of QoIs has selected for the occurrence of F129L mutations, which may contribute to loss of fungicide efficacy.     

Fusarium solani f. sp. passiflorae: a new forma specialis causing collar rot in yellow passion fruit

The aim of this study was to characterize a Fusarium population obtained from yellow passion fruit (YPF) with collar rot using pathogenicity, morphocultural characteristics and molecular tests. Pathogenicity and disease severity were assessed in six plant species: YPF, zucchini, tomato, bean, soya bean and cucumber. Potato dextrose agar medium (PDA) was used to determine mycelial growth at five temperatures (15–35°C). The colour produced by isolates was also determined on PDA at 25°C. Synthetic nutrient agar medium was used to evaluate: (i) type of mycelium and phialides; (ii) size, shape and number of septa from conidia; and (iii) production of chlamydospores and perithecia. Molecular tests consisted of sequencing the ITS–5·8S rDNA region and elongation factor 1α (EF‐1α) gene. The isolates caused large lesions on YPF, zucchini and tomato, with YPF having the highest mean disease severity and being the only one that showed wilt symptoms and death of the plant. Thus the isolates showed host specificity. Maximum mycelial growth occurred at 25°C and the predominant colour was bluish‐white. The isolates produced long phialides, dense aerial mycelium, oval microconidia with a mean size of 9·5 × 2·6 μm, macroconidia of 32·7 × 3·4 μm with 3·3 septa, and chlamydospores; only one isolate lacked perithecia. Phylogenetic trees of the ITS region and EF‐1α gene showed that isolates from YPF formed a distinct group within the F. solani group and the formae speciales of F. solani. It is proposed to name all isolates from YPF as F. solani f. sp. passiflorae.     

Pathogenic fungi involved in root rot of peas in the Netherlands and their physiological specialization

Research on root rot pathogens of peas in the Netherlands has confirmed the prevalence ofFusarium solani, F. oxysporum, Pythium spp.,Mycosphaerella pinodes andPhoma medicaginis var.pinodella. Aphanomyces euteiches andThielaviopsis basicola were identified for the first time as pea pathogens in the Netherlands. Other pathogens such asRhizoctonia solani andCylindrocarpon destructans were also found on diseased parts of roots. F. solani existed in different degrees of pathogenicity, and was sometimes highly specific to pea, dwarf bean of field bean, depending on the cropping history of the field.A. euteiches was specific to peas, whereasT. basicola showed some degree of physiological specialization.     

Survey on the prevalence of Rhizoctonia spp. in European soils and determination of the baseline sensitivity towards sedaxane

The prevalence of Rhizoctonia spp. in European soils was determined by analysing soil samples from 282 locations. Rhizoctonia spp. were found in 68% of these samples from France, Germany, the UK, Poland, Italy, Spain, Hungary and the Czech Republic. Samples from 136 locations were further analysed by pyrosequencing. Seventy‐six percent of the isolates were Rhizoctonia solani and 24% binucleate Rhizoctonia spp. Rhizoctonia solani anastomosis group (AG) 5 was detected most frequently (25%), followed by AG 9 (16%) and AG 4 (13%). For the binucleate Rhizoctonia spp., AG E was most prevalent (13%). Rhizoctonia cerealis was not detected in soil samples. Soil type or cropping history had no effect on the type of Rhizoctonia observed. Rhizoctonia solani AG 5 was the most frequently detected AG irrespective of the previous crop. The spectrum of AGs detected was similar for France, Germany and Poland but was significantly different for the UK (P = 0·0016). Finally, the baseline sensitivity towards sedaxane, a new active ingredient for seed treatment, was analysed for all isolates. The results indicate a low baseline sensitivity (average EC₅₀ of 0·028 p.p.m.) for all Rhizoctonia AGs. No difference in sensitivity was observed with the isolates obtained from different countries.     

Putative new species of the genus Dickeya as major soft rot pathogens in Phalaenopsis orchid production

Bacterial soft rots are a serious limitation to the production of orchids and other horticultural plants. Here, the characterization of causative bacteria isolated from Phalaenopsis orchids showing symptoms, from a commercial production site, is reported. The most commonly isolated bacteria were identified as Dickeya spp. Partial sequencing of 16S rDNA, fliC and dnaX showed diversity among the isolates and divided the isolates into two groups, with greatest similarity to previously reported undefined Dickeya lineages from orchids (UDL‐3 and UDL‐4). Two isolates (B16, S1) were sequenced using next‐generation sequencing, which has provided draft genomes of these two isolates for further studies (Ali? et al., 2015 ). Newly developed fliC‐based lineage‐specific quantitative real‐time PCR assays were used to distinguish among the lineages and to assess their relative abundances in diseased tissues. Virulence and aggressiveness comparison tests in vivo on Phalaenopsis orchids, potato plants and witloof chicory leaves indicated high virulence and extreme maceration potential of these novel Dickeya isolates, compared to a reference panel of other Dickeya spp. Pantoea cypripedii (formerly Pectobacterium cypripedii), which has previously been reported as a soft rot pathogen of orchids, was not detected, and isolates obtained from culture collections did not cause symptoms on artificially infected Phalaenopsis orchids.     

Morphological and molecular characterization of Fusarium spp. associated with yellowing disease of black pepper (Piper nigrum L.) in Malaysia

Yellowing disease is one of the most important diseases of black pepper (Piper nigrum L.). To characterize the pathogen(s) responsible for yellowing disease of black pepper in Malaysia, 53 isolates of Fusarium were collected from the roots of diseased black pepper plants and from rhizosphere soils from major growing areas in Sarawak and Johor. A total of 34 isolates of F. solani and 19 isolates of F. proliferatum were obtained and identified based on morphological characteristics and molecular techniques. DNA sequencing of the internal transcribed spacers (ITS1 and ITS2) and 5.8S ribosomal DNA regions was conducted to identify Fusarium species. Nucleotide sequence analysis of the ITS regions revealed that this molecular technique enabled identification of Fusarium at the species level as F. solani and F. proliferatum. In a pathogenicity test on 3-month-old black pepper plants, F. solani was pathogenic, but F. proliferatum was not. On the basis of morphology, DNA sequences and pathogenicity of the fungal isolates from the diseased plants, we showed that yellowing disease on black pepper is caused by F. solani     

Calcium and calmodulin may not regulate the disease resistance and pisatin formation responses of Pisum sativum to chitosan or Fusarium solani

No correlation was found between the chitosan or Fusarium solani-induced disease resistance responses in pea pod tissue and fluctuations in [Ca²⁺], inhibition of calmodulin, blockage of Ca²⁺ channels or host membrane leakage. Addition of exogenous Ca²⁺ 3 h before or after chitosan or F. solani treatments of pea pod tissue failed to alter the host response within 6 h, the time when the host actively resists both the compatible (F. solani f. sp. pisi) and incompatible (F. solani f. sp. phaseoli) macroconidia. Additionally, Ca²⁺ applied exogenously 3 h before or after chitosan significantly altered the level of UV-absorbing material released from the host tissue; however, it failed to affect the chitosan's ability to elicit phytoalexin formation by 24 h. Addition of exogenous Ca²⁺ 3 h before or after inoculation with either the compatible (f. sp. pisi) or incompatible (f. sp. phaseoli) fungi did not significantly change the host response by 24 h. The addition of EGTA or Ca²⁺ channel antagonists with the chitosan treatments also failed to significantly alter the chitosan-induced host response, thereby suggesting that chitosan probably does not function in the pea system by causing a Ca²⁺ influx into the host tissue which might then activate the host's resistance response. Inhibition of calmodulin related activities by calmidazolium failed to inhibit the chitosan or fungal induced host response. These results suggest that the response(s) induced in pea pod tissue by chitosan treatment or fungal inoculation may not be mediated by Ca²⁺, calmodulin or membrane leakage.