Conditions for Infection of Apple by Phytophthora syringae

The processes leading to Phytophthora fruit rot were divided into two main stages for the purposes of investigating the effects of temperature and duration of wet periods on pathogen development: oospore germination and infection of fruit by zoospores. It was found that the first stage was markedly affected by temperature over the range 10–20°C and required a wet period of 4–7 days. At 18 and 20°C, activation was low regardless of the length of the wet period. Once oospore germination (first stage) had occurred, free water was necessary for only a few hours for fruit infection (second stage) to occur, but the incidence of infection rose rapidly over the first 48 h, regardless of temperature over the range 10–20°C. From the data obtained, mathematical models were produced relating the incidence of Phytophthora fruit rot to the two weather variables. These models can be used to develop a weather‐based risk assessment system for the disease.     

Isolation of anti-fungal phenolic compounds from petioles of two Hevea brasiliensis (rubber) genotypes and their effect on Phytophthora meadii

Phenolic compounds were present in greater amounts in non‐infected petioles of genotypes of Hevea brasiliensis that are resistant to Phytophthora leaf disease than in genotypes that are susceptible. Phenolic compounds extracted from petioles of either susceptible (PB86) or resistant (RRIC100) genotypes, before or after infection with Phytophthora meadii, had anti‐fungal properties. Artificially infected petioles of PB86 had phenolic acids, triterpenoids or flavonoids, whereas healthy petioles contained only triterpenoids or flavonoids. However, healthy or infected petioles of RRIC100 contained only trace amounts of the above compounds and of vanillin (3‐methoxy‐4‐hydroxybenzaldehyde). Vanillin and umbelliferone (7‐hydroxycoumarin) were shown to suppress zoospore germination of P. meadii on glass slides and to inhibit its growth in pea broth and V‐8 juice agar. Vanillin was slightly more active than umbelliferone. Resistance of RRIC100 to Phytophthora was suspected as being related to the polymerisation of phenolic compounds to form lignin, which may suppress further spread of the pathogen's mycelium into healthy tissues. Formation of lignin from phenolic aldehydes as a barrier to disease spread may be a critical factor in resistance.     

Phylogenetic analyses and cross-infection studies of Phytophthora species infecting cacao and durian in South-Central Mindanao,Philippines

Phytophthora pathogens have contributed largely to the production losses of plantation crops such as cacao and durian. Correct identification and knowledge of the diversity of the pathogen is a requisite in the formulation of effective management strategies. Fifty-eight putative Phytophthora isolates were collected from cacao and durian orchards in South-Central Mindanao, Philippines, with 43 from cacao and 15 from durian. Molecular identification of isolates based on DNA sequences of the internal transcribed spacer (ITS), partial β-tubulin (β-tub) and mitochondrial gene (cox2+spacer) regions combined with morphocultural characterization identified 57 Phytophthora palmivora isolates from cacao and durian typically showing variations in colony morphology and sporangial shape. However, these variations were not phylogenetically informative. One Phytophthora meadii (CDDS-P3) causing pod rot of cacao was phylogenetically isolated from P. palmivora. Phylogenetic analysis of isolates using combined ITS, β-tubulin (β-tub) and mitochondrial (cox2+spacer) DNA regions showed an intraspecific variation within P. palmivora. Representative isolates of P. palmivora used for cross-inoculation showed more aggressiveness on its original host with higher lesion diameter and less aggressiveness on its potential hosts with smaller lesion diameter on artificially inoculated detached plant part. All Phytophthora isolates were tested pathogenic to cacao, durian and its potential hosts (rubber and coconut); therefore, cross-infection in the field is possible in an intercrop system. Implementation of combined control strategies should be done against highly diverse pathogens such as P. palmivora and the potentially destructive pathogen P. meadii, which was tested pathogenic and virulent to its original (cacao) and potential hosts (coconut, durian and rubber).     

Biological control of fruit-rot of jackfruit by rhizobacteria and food grade lactic acid bacteria

Fungal species causing fruit rot of jackfruit have been isolated from seven different locations of Birbhum and Burdwan districts of West Bengal, India. Each isolate showed more or less similar microscopic characteristics. A representative strain VBAM1, isolated from a severely infected jackfruit was identified as Rhizopus stolonifer by 28S rDNA sequence homology. Increased reducing sugar content in pectin broth indicates pectinase production by the pathogen. The pathogen was not inhibited by ⩾500 μg/ml of Mancozeb and Bavistin. Copper oxychloride, Blytone 50% a.i. showed antifungal activity at comparatively lower concentration (200 μg/ml). Two rhizospheric bacterial strains, Burkholderia cenocepacia VBC7 and Pseudomonas poae VBK1, and three different strains of Lactococcus lactis subsp. lactis can produce significant zones of inhibition against the pathogen in dual culture overlay plates. They induced mycelia breakage of pathogen as evidenced from scanning electron micrographs. When applied to whole plants, the strains reduced or prevented disease and when applied postharvest to Rhizopus inoculated fruit delayed and/reduced disease incidence. These agents were also re-isolated from the applied surfaces and survived for long time when mixed with suitable carrier base indicating stability in a formulation over time.     

Fusarium incarnatum is associated with postharvest fruit rot of muskmelon (Cucumis melo)

Postharvest fruit rot was observed on muskmelon (Cucumis melo) on market shelves at a melon farm in the city of Hatyai, Songkhla, Thailand. Diseased muskmelon fruit displayed cotton-like mycelia on wounds and had brown to dark brown internal tissue. Based on morphological characteristics and molecular analysis of internal transcribed spacer (ITS) and translation elongation factor 1-α (TEF1-α) DNA sequences, the fungal pathogen was identified as Fusarium incarnatum. This species belongs to the Fusarium incarnatum-equiseti species complex (FIESC). A pathogenicity test was conducted to verify Koch's postulates, and F. incarnatum was observed to cause fruit rot on muskmelon; symptoms of the disease were similar to those seen in the field. However, only artificially wounded melons became infected, suggesting that F. incarnatum is an obligate wound-infecting pathogen. To our knowledge, this is the first report of F. incarnatum causing fruit rot of muskmelon in Thailand.     

Morphological and Molecular Characterization of Pomegranate Fruit Rot Pathogen, <Emphasis Type="Italic">Chaetomella raphigera</Emphasis>, and its Virulence Factors

A new fungal pathogen was isolated from rotten pomegranates collected from the orchards of different parts of Maharashtra. The pathogen was morphologically identified as Chaetomella raphigera followed by sequencing of ITS and D1/D2 hypervariable region of LSU (28S) of rRNA gene. The pathogen produced pectinase, cellulase, xylanase and protease in liquid medium at a concentration of 71, 13.8, 54.3 and 7 U/ml respectively. Enzyme activity was also determined during pathogenesis in the tissues artificially infected by C. raphigera. Xylanase activity was maximum (25.1 U/g) followed by pectinase (19.2 U/g) and cellulase (1.5 U/g), whereas, protease activity was unnoticed. There was significant correlation (P < 0.05) between disease rating scale and pectinase, xylanase and cellulase activity in infected tissues. This indicates the simultaneous production of hydrolytic enzymes that aids in necrosis of fruit tissues. The elevated levels of these enzymes in infected tissues as compared with control suggest their possible role in pathogenesis. Thus, pectinase, cellulase and xylanase produced by C. raphigera acts as major virulence factors in the development of fruit rot in pomegranates. This is a first report of fungal fruit rot caused by C. raphigera in pomegranate.     

Modeling the spread of Phytophthora

We consider a model for the morphology and growth of the fungus-like plant pathogen Phytophthora using the example of Phytophthora plurivora. Here, we are utilizing a correlated random walk describing the density of tips. This random walk incorporates a delay in branching behavior: newly split tips only start to grow after a short while. First, we question the effect of such a delay on the running fronts, for uniform- as well as non-uniform turning kernels. We find that this delay primarily influences the slope of the front and therewith the way of spatial appropriation, and not its velocity. Our theoretical predictions are confirmed by the growth of Phytophthora in concrete experiments performed in Petri dishes. The second question addressed in this paper, concerns the manner tips are interacting, especially the point why tips stop to grow “behind” the interface of the front, respectively in confrontation experiments at the interface between two colonies. The combination of experimental data about the spatially structured time course of the glucose concentration and simulations of a model taking into account both, tips and glucose, reveals that nutrient depletion is most likely the central mechanism of tip interaction and hyphal growth inhibition. We presume that this is the growing mechanism for our kind of Phytophthora in infected plant tissue. Thus, the pathogen will sap its hosts via energy depletion and tissue destruction in infected areas.     

Novel bacterial endophytes from Hevea brasiliensis as biocontrol agent against Phytophthora leaf fall disease

Bacterial endophytes offer control against many diseases of crop plants as potential biocontrol agents. Antagonistic bacterial endophytes acting against Phytophthora meadii have been screened from leaf, petiole and root tissues of Hevea brasiliensis. Six bacterial endophytes could exhibit more than 50 % inhibition of P. meadii, among which EIL-2, from disease-free zones showed a maximum of 62.5 % inhibition. The isolate EIL-2 was characterized as Alcaligenes sp. and the other isolates were identified as Pseudomonas aeruginosa. 16S rDNA sequence analysis showed that there existed genetic variation among the five isolates of P. aeruginosa from different tissues of the plant indicating the tissue type adaptation of the isolates. Dual culture technique with endophyte EIL-2 completely arrested the growth of P. meadii when inoculated prior to pathogen. The bioassay with EIL-2 in H. brasiliensis clones, RRII 105 showed 43 % reduction of lesion size on infected leaves whereas in RRIM 600 it was only 30 %.     

Development of an Improved Isolation Approach and Simple Sequence Repeat Markers To Characterize Phytophthora capsici Populations in Irrigation Ponds in Southern Georgia

Phytophthora capsici, the causal agent of Phytophthora blight, is a major concern in vegetable production in Georgia and many other states in the United States. Contamination of irrigation water sources by P. capsici may be an important source of inoculum for the pathogen. A simple method was developed in this study to improve the efficiency of recovering P. capsici from fruits used as baits in irrigation ponds. In contrast to direct isolation on agar plates, infected fruit tissues were used to inoculate stems of pepper seedlings, and the infected pepper stems were used for isolation on agar plates. With isolation through inoculation of pepper stems, the frequency of recovering P. capsici from infected eggplant and pear fruits increased from 13.9% to 77.7% and 8.1% to 53.5%, respectively, compared with direct isolation on agar plates. P. capsici was isolated from seven out of nine irrigation ponds evaluated, with most of the ponds containing both A1 and A2 mating types and a 4:5 ratio of A1 to A2 when isolates from all ponds were calculated. All P. capsici isolates were pathogenic on squash plants, and only a small proportion (8.2%) of the isolates were resistant or intermediately sensitive to mefenoxam. Simple sequence repeats (SSRs) were identified through bioinformatics mining of 55,848 publicly available expressed sequence tags of P. capsici in dbEST GenBank. Thirty-one pairs of SSR primers were designed, and SSR analysis indicated that the 61 P. capsici isolates from irrigation ponds were genetically distinct. Cluster analysis separated the isolates into five genetic clusters with no more than two genetic groups in one pond, indicating relatively low P. capsici genetic diversity in each pond. The isolation method and SSR markers developed for P. capsici in this study could contribute to a more comprehensive understanding of the genetic diversity of this important pathogen.Phytophthora capsici, the causal agent of Phytophthora blight, is a widespread and destructive plant pathogen that causes root rot, crown rot, fruit rot, and foliar blight on many economically important crops in the United States and throughout the world (1). A number of important vegetable crops are susceptible to this pathogen, including peppers, squash, cucumber, watermelon, cantaloupe, zucchini, eggplant, pumpkin, tomatoes, and snap beans. The pathogen causes significant yield reductions and quality losses to vegetable industries and has become a major concern in vegetable production in the United States in recent years. The efficacies of current strategies for management of the disease are limited. No single fungicide has consistently and effectively suppressed losses caused by P. capsici epidemics. While fungicides containing the active ingredient mefenoxam provide some level of control of P. capsici, mefenoxam-resistant isolates that challenge the usefulness of the compound have developed (3, 8).It is critical to understand the ecology and epidemiology of P. capsici in order to design more effective disease management strategies. Studies conducted in recent years indicate that P. capsici survives in irrigation water in the United States, and irrigation water may serve as an important inoculum source. Roberts et al. (14) reported that P. capsici was isolated from tailwater (surface runoff water) in Florida using water filtration and lemon leaf baiting techniques. Gevens et al. (3) used pear and cucumber fruits as baits and isolated P. capsici from irrigation water sources in Michigan. It was unknown, however, if irrigation water sources in Georgia could be significant sources of primary inoculum. Earlier studies using water filtration or direct isolation from water and bottom sediment did not identify P. capsici in surface irrigation ponds in Georgia (16).Since surface water can be a significant source of P. capsici, it is critical to use appropriate methods to isolate the pathogen from irrigation water and to facilitate characterization of the isolates. Fruit, especially pears, is often used as bait to recover Phytophthora spp. from water (3, 21). In comparison to water filtration, the baiting technique is easier and less labor intensive. However, direct isolation from infected fruit bait is often hampered by other microorganisms. Isolation of Phytophthora spp. is often affected by Pythium spp. that overgrow fruit and agar plates. Hence, development of a more efficient isolation method is needed to increase the frequency of P. capsici recovery to facilitate the detection and characterization of isolates associated with water sources.The objectives of this study were to develop an efficient method to isolate P. capsici from irrigation ponds in southern Georgia and to develop simple sequence repeat (SSR) markers to analyze the genetic diversity of P. capsici populations in irrigation ponds. SSRs are tandemly repeated motifs of 1 to 6 bases found in the nuclear genomes of all eukaryotic organisms and are often abundant and evenly dispersed (7). They are highly polymorphic, multiallelic, and codominant and are believed to be a more efficient marker system than restriction fragment length polymorphisms and randomly amplified polymorphic DNAs (18, 23). SSR markers have been derived from publicly available expressed sequence tags (ESTs) of a few plant pathogens, including Phytophthora infestans, Phytophthora sojae, and Magnaporthe grisea (5, 10, 23); however, no SSRs for P. capsici have been developed. Development of EST-SSR markers may provide an effective molecular marker system for analysis of genetic variation within P. capsici populations.     

First morphogenetic identification of <i>Fusarium solani</i> isolated from orange fruit in Egypt

Losses due to postharvest decay may occur at any time during postharvest handling, from harvest to consumption affecting the produce quality and quantity. Accurate identification of the pathogen causing postharvest disease is essential to the selection of an appropriate disease control approach. Nine isolates of Fusarium recovered from orange fruit were identified as Fusarium solani. The fungus is involved with fruit decay. The obtained cultures were purified and grown on potato-dextrose agar (PDA), malt yeast agar (MYA), and Czapek's nutrient media (CNM) under light for identification. A pathogenicity test was carried out to fulfil Koch's postulates. The pathogen could only enter ripe orange fruit through wounds and cracks causing the rot disease. The identification of the fungal isolates was confirmed to be F. solani by DNA sequencing, which was 99 to 100% homologous to those deposited in the Gen- Bank. The identity of nine fungal isolates was confirmed to be F. solani by DNA sequencing of the internal transcribed spacer (ITS) rDNA region (GenBank Accession Nos. DQ486874 to DQ486881 and KC758879). To our knowledge, this is the first morphogenetic identification of F. solani isolated from orange fruit in Egypt.     

Supplementary Antimold Activity of Phosethyl AL, A New Brown Rot Fungicide for Citrus Fruits

Phosethyl Al (= Aliette) is a new and promising postharvest fungicide against brown rot of citrus fruits. Its effectiveness in controlling green mold (Penicillium digitaum), the major citrus fruit pathogen, was studied on artificially inoculated fruit and compared with that of sodium o-phenylphenate (SOPP) and thiabendazole (TBZ). In vitro and in vivo experiments have shown that phosethyl Al is capable — although to a lesser extent than SOPP and TBZ — of inhibiting growth of P. digitatum and reducing the incidence of decay of citrus fruits, caused by a wild-type, and apparently also by a benzimidazole-resistant strain of this fungus. The influence of times of treatment and modes of application (including temperature and duration of treatment, rinsing of the fruit) on the effectiveness of phosethyl Al, SOPP and TBZ in controlling green mold was studied and discussed. It is concluded that, under conditions of natural infection, phosethyl Al can be used for the postharvest control of Phytophthora-caused brown rot without any apparent increase in the incidence of green mold in the treated fruits.     

Studies on the Biological Control of Phytophthora cryptogea Pethybr. et Laff. II. Effectiveness of Trichoderma and Gliocladium spp. in the Control of Phytophthora Foot Rot of Gerbera

Bip T containing about 10⁹ spores of Trichoderma viride, applied to peat 10 days before inoculation of substrate with Phytophthora cryptogea, effectively controlled Phytophthora foot rot of gerbera. The biocide in dosage of 300 and 600 g/m³ inhibited the development of the pathogen in substrate. The other potential antagonists T. hamatum and T. viride applied into peat at a dosage of 600 g/m³, decreased Phytophthora foot rot development.     

Pectobacterium carotovorum subsp. brasiliense Causing Soft Rot on Paprika in Korea

During summer 2011 in South Korea, severe fruit rot of paprika was observed, causing severe economic losses in paprika production. Symptoms of fruit and pedicel decay were consistent with symptoms caused by Pectobacterium carotovorum subsp. brasiliense (Pcb) as recently described in Brazil, the United States, Israel and South Africa. Physiological analysis and pathogenicity test of strains isolated from paprika fruit revealed that the pathogen was the bacterium Pcb. Sequencing and phylogenetic analysis of the 16S rDNA and partial 16S–23S rDNA intergenic spacer region confirmed that the isolates were Pcb. This is the first report of Pcb in Korea, which has a significant economic impact on Korean paprika production.     

Sources of crown rot (Phytophthora cactorum) infection in strawberry and the effect of cold storage on susceptibility to the disease

Cold-stored plants of strawberry cultivars Tamella, Cambridge Favourite and Redgauntlet were more susceptible to pathogenic isolates of Phytophthora cactorum than similar plants which had not been cold-stored. Indigenous nonpathogenic isolates of P. cactorum did not cause crown rot in cold-stored plants, although a small number of symptomless latent infections occurred. The majority of P. cactorum isolates causing crown rot symptoms were taken from infected strawberry crowns, although two isolates from gooseberry plants, but of uncertain origin, were also pathogenic. Outbreaks of crown rot in areas with no previous history of the disease therefore probably result from the importation of non-indigenous inoculum with planting material. Assessments of the timing of infection in relation to cold storage revealed that a high incidence of death in the cold store and chronic wilt symptoms on planting from the store resulted from initiating symptomless infections prior to cold storage. However, infection during the period immediately after cold storage resulted in rapid wilt symptoms of Phytophthora crown rot. When plated in sterile distilled water for 24 h, pieces of tissue from infected plants which had died during cold storage produced large numbers of sporangia and zoospores. This indicates that such plant material could provide a potent source of inoculum for infections in the post storage thawing environment. It is proposed that a combination of heightened host susceptibility resulting from cold storage and the presence of scatted latent infections or infected debris among the plants could result in a sudden, large scale appearance of crown rot, as sometimes is seen with cold-stored plantings of strawberries.     

Genome-wide identification of laccase gene family in three Phytophthora species

Phytophthora spp. is a primary pathogen in oomycete, causing economically and environmentally devastating epidemics of plants. Laccases have been found in all domains of life but have not been reported in oomycte. In this paper, laccase genes of Phytophthora spp. were identified in three genomes (Phytophthora capsici, Phytophthora sojae and Phytophthora ramorum). 18 laccase genes were identified in total, including four in P. capsici genome, six in P. sojae genome and eight in P. ramorum genome. Most of the predicted gene models shared typical fungal laccase character, possessing three conserved positions with one cysteine and ten histidine residues at these positions. Phylogenetic analysis illustrated that laccases from Phytophthora clustered into four clades, while fungal laccases clustered together. The results provided the theoretical ground for new hypotheses about the roles laccases in oomycetes and may guide the future research of these enzymes.     

Phytophthora chrysanthemi sp. nov., a new species causing root rot of chrysanthemum in Japan

A new species of Phytophthora was isolated from stem and root rot of chrysanthemum in the Gifu and Toyama prefectures of Japan. The species differs from other Phytophthora species morphologically, and is characterized by nonpapillate, noncaducous sporangia with internal proliferation, formation of both hyphal swellings and chlamydospores, homothallic nature, distinctive intercalary antheridia, and funnel-shaped oogonia. The new species can grow even at 35°C, with an optimum growth temperature of 30°C in V8 juice agar medium. In phylogenetic analyses based on five nuclear regions (LSU rDNA; genes for translation elongation factor 1α, β-tubulin, 60 S ribosomal protein L10, and heat shock protein 90), the isolates formed a monophyletic clade. Although the rDNA ITS region shows a high resolution and has proven particularly useful for the separation of Phytophthora species, it was difficult to align the sequences for phylogenetic analysis. Therefore, ITS region analysis using related species as defined by the multigene phylogeny was performed, and the topology of the resulting tree also revealed a monophyletic clade formed by the isolates of the species. The morphological characteristics and phylogenetic relationships indicate that the isolates represent a new species, Phytophthora chrysanthemi sp. nov. In pathogenicity tests, chrysanthemum plants inoculated with the isolates developed lesions on stems and roots within 3 days, and the symptoms resembled the ones originally observed. Finally, the pathogen’s identity was confirmed by re-isolation from lesions of infected plants.     

A novel Fomitiporia species associated with esca on grapevine in South Africa

Esca disease of grapevine is a complex trunk disease consisting of several symptoms, one of which, white rot, has been found to be caused by various basidiomycetes within the order Hymenochaetales. During recent surveys of esca-related pathogens in South African vineyards, several unidentified basidiomycetes were isolated from white rot occurring in diseased vines. A new Fomitiporia species, F. capensis, is described based on fruit body morphology and combined internal transcribed spacer (ITS) and large-subunit (LSU) phylogeny, where it forms a clearly delineated and well-supported clade. Morphologically, F. capensis is similar to F. punctata in that both species essentially lack setae. Fomitiporia capensis, F. punctata and F. aethiopica produce similarly sized basidiospores but differ in terms of host range, pore size and, possibly, fruit body shape. Phylogenetically, F. capensis appears to be related to F. tenuis, though morphologically, the species differ significantly in that F. tenuis has smaller pores and smaller basidiospores. Fomitiporia capensis was found to occur widely as vegetative mycelium throughout the Western Cape Province, though fruit bodies were scarce in comparison. A vineyard with fruit bodies was also found in Limpopo in the north east of the country. Fruit bodies were found growing on the underside of the cordon of living vines displaying external symptoms typically associated with esca, or general decline and dieback symptoms together with internal white rot.     

Expression of phenolics and defence-related enzymes in relation to red root rot disease of tea plants

Red root rot caused by Poria hypolateritia is a dreadful disease in tea plant due to sudden death of bushes. In response to fungal pathogen, variation in the defence-related enzymes was investigated. The infected tea root was undertaken to study about various defence-related and pathogen-related enzymes. The infected root, as a prime response to disease attack, was subjected to the analysis of phenolics, phenylalanine ammonia lyase, tyrosine ammonia lyase, peroxidase, polyphenol oxidase, catalase, chitinase, β-1,3-glucanase and protease were assayed. The results on assay of defence-related enzymes revealed that the activity was significantly higher in infected roots when compared with healthy roots. Phenolics were accumulated more in infected roots. The sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis further confirmed the presence of induced pathogenesis-related proteins in the infected root tissues. The activity of all enzymes was increased up to threefold amount when compared with normal ones. The accumulation of defence enzymes in plants revealed the virulence of root pathogen in stimulating induced systemic resistance of tea plants and phytopathogenicity causing pathogenesis. This study exemplify to recognise underlying processes in causing infection and to identify the existence of host–pathogen relationship.     

Microscopic and polyclonal antibody-based detection of yellow leaf disease of arecanut (Areca catechu L.)

Phytoplasma, the pathogen of yellow leaf disease (YLD) of arecanut (Areca catechu L.) was detected by transmission and scanning electron microscopy. Tissues of YLD affected palms contained phytoplasmas in the phloem sieve elements, but not in symptomless healthy palm tissues. Phytoplasma was purified from tissues of diseased palms employing percoll density gradient centrifugation and confirmed by transmission electron microscopy. Using the purified phytoplasma preparation, a polyclonal antiserum was raised in rabbits and used for standardisation of agar gel double diffusion (Ouchterlony) test and DAC-ELISA. Clear precipitin line was observed in Ouchterlony test between the antigen from diseased palms and the pathogen-specific antibodies after 48 h incubation and only undiluted antiserum showed best result in the test. However, in ELISA, 1:10 antigen dilution and 1:400 pathogen-specific antibodies dilution produced sensitive detection of the pathogen with a difference of >3.5 times absorption values between healthy and diseased samples. The results thus confirmed the ability of antiserum to distinguish healthy and infected plants and utility of ELISA for effective diagnosis of YLD.