The isolated spp of Trichoderma was found to be Trichoderma harzianum and Trichoderma viride. Cluster III and IV contained two isolates each. The result of the phylogenetic analysis based on the Tef1 gene sequences of 20 Trichoderma isolates is shown in Fig. The mechanisms used by these two endophytic strains against the pathogen were evaluated by cryo-scanning electron microscopy. (2017) described several Trichoderma isolates that developed profuse fluffy mycelium and two to three fine defined concentric mycelium (white) and conidia (green) rings. In this study, 23 strains of Trichoderma were isolated from terrestrial and marine environments and identified using a polyphasic approach of morphological characterization and ITS gene sequence analysis. . The plates were then incubated at 281C. Principle component analysis (PCA) was performed using XLSTAT software. Turn the Petri dish on end. In vitro antagonistic potential of the biocontrol agent was evaluated against Fusarium oxysporum f. sp. Diversity and antagonistic potential of, Srivastava AK, Singh RN, Kumar S, Kashyap PL, Arora DK. Aspergillus can be found in a variety of environments throughout the world given that their growth is largely determined by the availability of water. However, ERG-1 gene also showed heterogeneity among some antagonistic isolates and indicated the possibility of occurrence of squalene epoxidase driven triterpene biosynthesis as an alternative biocontrol mechanism in Trichoderma species. http://www.austin.cc.tx.us/microbugz/03morphology.html, accessed January 14, 2005. After incubation, it was again centrifuged at 13,000rpm for 20min at 4C temperature. Methods based on such repetitive elements have also been used for studying the diversity in the ecosystem, presenting the phylogenetic relationship between strains and discriminating between microorganisms those are genetically close to each other (Rai et al. <>endobj Morphology and life cycle of Trichoderma viride (green mold) Depending on the substrate on which it grows, T.viride appears to be a light greenish or yellowish mold with somewhat granular or powdery conidia without the formation of concentric rings on the colony (Fig.1)(2). Phytohormone profiles induced by, Mohapatra BR, Broersma K, Mazumder A. 34 isolates of Trichoderma were obtained from various geographical locations in India and were subjected to phenotypic evaluation. If a colony displays a greater growth rate than the other colonies, a curve is likely to be observed in the contact zone. Being kept in one place, the resulting cells have accumulated to form a visible patch. Biochemical and metabolic profiles of, Malmierca MG, Cardoza RE, Alexander NJ, McCormick SP, Collado IG, Hermosa R, et al. You may print and distribute up to 200 copies of this document annually, at no charge, for personal and classroom educational use. Plant defense activation and management of tomato root rot by a chitin-fortified, Solanki MK, Kumar S, Pandey AK, Srivastava S, Singh RK, et al. Form - What is the basic shape of the colony? 0000273901 00000 n x][6~wC\IfSM]{1+ZOw %DsJe ~ht7 u^&tuYr|muvy!~LL7?u|zGD^O/_0 i)qO/_|vzIw7}20E2,{{w[q7OO?|gi6V"O=_\lcSQu-!-?JdXYhQw)zahcZ|C;#jfgHoga uPW1bt-*>7IO'G0~@{>N ':=&+lsJ(9W^e/X*7. are rapidly growing and form hyaline hyphae that give rise to yellow-green to green patches of conidia formed on clusters . Based on the sequence analysis of EF1 gene, the 20 antagonistic isolates were divided in five species: T. harzianum, T. koningii, T. asperellum, T. virens and T. viride. Mounts were examined under the microscope to observe the morphological characteristics of conidia and conidiophores. All the two species were distinct from each other in the characteristics such as mycelial growth rate, colony appearance, shape of conidia and conidiophores, and branching . 2005). Principal component analysis (PCA) based on first and second coordinates showed a maximum Eigen value of 9.758 and minimum value of 0.017 with a percentage variation of 48.71 and 13.16, respectively (Fig. %%EOF Morphological characterization. All RAPD, ERIC, REP and BOX-PCR reactions were repeated to ensure validity of results. Common physical characteristics of bacteria colonies are listed and separated into 3 categories. The comparison of the genetic structure between antagonistic Trichoderma isolates was carried out by molecular (RAPD, REP, ERIC and BOX markers), and biochemical (production of cell wall degrading enzymes) markers. endobj Two examples of molds are shown below: Moss green colonies, a white cloud, or a ring of spores can be attributed to the growth of Aspergillus, which is common in such fungal infections as athlete's foot. The purified isolates were identified with their morphology and molecular . When printing this document, you may NOT modify it in any way. http://www.shroomery.org/images/23418/Aspergillus_nidulaus.jpg, accessed January 14, 2005. This study provides information on post-treatment procedures that . 2015). 2004; El_Komy et al. 2. The number of bands in the amplification profile was 182, and their size was found to vary from 250 to 3000bp among these isolates (Fig. All . Austin Community College. A plethora of antagonistic Trichoderma isolates have been identified by several researchers from different places around the world, having history of varied climate, soil type, cropping system, etc., which differ in their innocuousness and efficacy as biocontrol agents (Sharma et al. hb```b``e`c` @1v+ C{PFs9X%Ops600|yfMrN81 xi{ISy./K}Q1ls82nNl84EgQ.JaWM>7NC>L 'Yd6^[2.T Singh RK, Kumar DP, Singh P, Solanki MK, et al. What are the Chances? 0000168795 00000 n phialide, conidiophore, chlamydospores, and the colony growth patterns of strain GD040 were observed (Xue et al., 2021). being able to grow at 45 C (113 F). All the PCR reactions were carried out in 25l reaction mixture containing 5 Gitschier buffer, 50ng DNA template, 2mM MgCl2, 0.25mM dNTP mixture and 0.25M each of primer, and one unit of Taq Polymerase (Bangalore Genie, India). PCR amplification of squalene epoxidase (ERG1) gene, showing ~500bp amplicon in Trichoderma isolates having distinct geographical lineages. Copyright 2002-2023 Science Buddies. Sudheer Kumar, Email: moc.liamg@raci.reehdus. (11) The Shroomery. A 5 mm diameter disc using a sterile cork borer was cut from the actively growing edge of a fresh colony (before the The use of Trichoderma isolates with efficient antagonistic activity represents a potentially effective and alternative disease management strategy to replace health hazardous chemical control. 2. The Trichoderma strains were tested for their tolerance and biosorption of nickel. (12) ETH Life International. are termed the colony morphology. Observation of growth pattern on different media and colony properties led us to classify the . The genus Trichoderma has five species; Trichoderma harzianum, Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma pseudokoningii, and Trichoderma viride. We use cookies and those of third party providers to deliver the best possible web experience and to compile statistics. Mating type genes and genetic markers to decipher intraspecific variability among, Kashyap PL, Rai S, Kumar S, Srivastava AK. 0000004455 00000 n Trichoderma includes a variety of fungal species used for biological control of phytopathogenic fungi. 2007; de los Santos-Villalobos et al. Trichoderma spp. PCA analysis explained 31.53, 61.95, 62.22 and 60.25% genetic variation among Trichoderma isolates based on RAPD, REP-, ERIC- and BOX element analysis, respectively. The fungus T. harzianum was grown . Species of Clonostachys and Trichoderma isolated from the surface of bananas were highly antagonistic to crown rot-causing fungal pathogens of banana, . The .gov means its official. Then . Hypocrea spp. Colony morphology observations are an important pre-step in the identification and further use of cultivated colonies. Trichoderma species are a group of fungi which is widely distributed in major terrestrial ecosystems; they are also commonly used as biocontrol agents for many plant diseases. 0000011611 00000 n The characteristics of a colony (shape, size, pigmentation, etc.) (8) Biology at the University of Cincinnati Clermont College. 0000014182 00000 n From the front, the color is white. 2014; Galarza et al. In present study, Trichoderma isolates were categorized on the basis of description and keys given by Gams and Bissett (1998). Isolates UNT60, UNT68, NAT70, DET89, HAT96, UNT38, UNT13 and UNS30 grew faster (13.3mmday1) than other isolates. The media 0000002556 00000 n 2015). Washington State University. 0000008456 00000 n 0000168067 00000 n The supernatant was discarded and pellet was washed with 70% ethanol. 4 0 obj Srivastava AK, Singh P, Singh RK, Kashyap PL, et al. The isolates were further screened for their antagonistic potential against the pathogen on PDA by measuring the relative growth rates as a function of the incubation period. However, further experiments are needed to validate the role of squalene epoxidase driven triterpene biosynthesis in biocontrol mechanisms of tested isolates. 2015). 4 0 obj In general, both techniques were found to produce reproducible results especially with purified genomic nucleic acid as a template, and when the primer concentration and composition of buffer were strictly controlled. Ethidium bromide (0.25mgml1) was used as an intercalating agent. Seven new species of the genus Trichoderma belonging to the Viride clade are described from soil samples of different regions in China, and named as T. adaptatum, T. beijingense, T. bifurcatum, T. mangshanicum, T. shaoguanicum, T. tardum and T. vulgatum. Solanki MK, Singh N, Singh RK, Singh P, Srivastava AK, Kumar S, Kashyap PL, Arora DK. The Trichoderma isolates were initially sorted according to morphology and based on the translation elongation factor 1- gene sequence similarity, . Conidiophores are hyaline, branched, and may occasionally display a pyramidal arrangement. In this context, twenty isolates were obtained from tomato rhizosphere and evaluated by their antagonistic activity against four fungal pathogens (Fusarium oxysporum f. sp. Septate hyaline hyphae, conidiophores, phialides, and conidia are observed. However, globose and sigmoid or hooked phialides were also observed in two (UNT68 and ALT73) and one isolate (NAT03), respectively (Table2). 0000282422 00000 n (2013a). 0000008727 00000 n Morphological characteristics of four Trichoderma isolates and two endophytic Fusarium isolates. It does not have the typical structure of a bacterium, since Streptomyces coelicolor is a gram positive bacterium, whose habitat is widespread throughout the world. 2013; Cardoza et al. ergosterol) from tomato rhizosphere. Opacity - For example, transparent (clear), opaque, translucent (almost clear, but distorted vision, like looking through frosted glass), iridescent (changing colors in reflected light), etc. Similarly, maximum -1,3 glucanase production was observed in UNS30 followed by DET94 (Table1). morphology, physiology, and colony characteristics distinguish type I and type II as different species . (10) The Shroomery. Colonies of Trichoderma grow rapidly and mature in 5 days. The rate at which they grow is largely determined by the temperature range in . Trichoderma crassum was found to be a sister species to T. virens, based on molecular sequences and phenotypic data. Trichoderma morphology. These patches may sometimes form concentric rings. 2009; Baszczyk et al. Shalini Rai, Email: moc.liamg@90orciminilahs. PLK participated in the design of the study, performed statistical analysis and helped to draft the manuscript. Supernatant was taken into another Oakridge tube and an equal volume of phenol:chloroform:isoamyl alcohol (25:24:1) was added with slow inversion. A perusal of the PCA analysis revealed that eight isolates (HAT96, UNT68, DET94, UNT60, UNT64, NAT69, DET02, DET89 and UNT09) formed a major cluster (cluster IV), while three isolates were grouped in cluster II (UNS28, UNS63 and UNT38) and IV (UNT70, NAT03 and ALT73). Mycoparasitism studies of, Radjacommare R, Venkatesan S, Samiyappan R. Biological control of phytopathogenic fungi of vanilla through lytic action of, Rai P, Sharma A, Saxena P, Soni AP, et al. Careers, Unable to load your collection due to an error. The phylogenetic relationships of these species were inferred based on partial RPB2 and EF-1 DNA sequence data and phenotypic characteristics, including teleomorph, anamorph, colony and growth rates. The structural and functional analysis of genes involved in the synthesis of ergosterol especially intermediates, such as squalene could provide additional strategies to improve the ability of biocontrol of the Trichoderma strains. with a volumetric pipette at a concentration of 1 10 8 colony-forming units (CFU) per mL directly into the soil. Identification of loci and functional characterization of trichothecene biosynthetic genes in the filamentous fungus, Cardoza RE, Malmierca MG, Gutirrez S. Overexpression of. These results are consistent with the findings of Markovich and Kononova (2003). 2011). VL tamatar 4) rhizosphere (Table1). The mixture was centrifuged at 13,000rpm for 15min at 4C to pellet the mycelium. The soil particles attached to roots were carefully collected after uprooting plants, stored at 4C and processed within 24h of collection. The number of bands in the amplification profile was 350, and their size was found to vary from 270 to 3000bp among these isolates (Fig. 2015). 2010; Solanki et al. For this, the pathogenic fungi were obtained from National Agriculturally Important Microorganisms Culture Collection (NAIMCC), NBAIM, Mau, Uttar Pradesh. Trichoderma longibrachiatum and Trichoderma viride may also produce chlamydospores. They reported that the mycoparasitic capacity of various species and isolates of Trichoderma differs. The authors declare that they have no competing interests. HHS Vulnerability Disclosure, Help The white areas do not contain spores, while the green areas are covered with dense masses . Species identification if based on only morphology, may lead to sometimes erroneous, uncertain or remains unclear due to overlapping characters. The phialides may be solitary or arranged in clusters. 0000005668 00000 n Before Despite the commercial successes of these biocontrol agents, the major limitations remain their restricted efficacy and inconsistency under field conditions. The phialides were nine-pin shaped and their size varied between 3.913.71.72.9 to 7.015.02.03.0m in seventeen isolates. Under such situations, it is valuable to establish patterns of gene flow, as well as to develop a fingerprint of Trichoderma isolates. Trichodiene production in a, Markovich NA, Kononova GL. 2009; Krmczi et al. For example, circular, filamentous, etc. (2003). Trichoderma species were isolated from different ecological habitats in Nuwara Eliya district using pour plate technique and five isolates (T1, T2, T3, T4 and T5) were recognized using colony . Surface - How does the surface of the colony appear? Each distinct circular colony should represent an individual bacterial cell or group that has divided repeatedly. http://tea.rice.edu/Images/stoyles/stoyles_pinkJPG.JPG.jpg, accessed January 14, 2005. For morphological analysis, isolates were grown on PDA at 281C for 57days. Terpene compounds (e.g., ergokonins and viridins) are involved in the biocontrol process due to their antifungal properties (Malmierca et al. 0000005215 00000 n Although it is commonly considered as a contaminant, Trichoderma may cause infections in presence of certain predisposing factors. The size of RAPD fragments ranged 2502500bp. 0000009629 00000 n 1 (A-C): Trichoderma asperellum CA-03/9840 strain in PDA medium and microscopic image. Umbonate colonies have a knob-like central elevation. Plimmer JR. Regulatory problems associated with natural products and biopesticides. 0000067697 00000 n W E L C O M E 2. For this pre-treatment, fresh mycelium and the agar medium underneath are frozen at 80 C and lysed mechanically. <> Thermocycling was run with the following settings: heating at 94C (5min); then 35 cycles of 94C (30s), 60C (30s) and 72C (90s). 3971 0 obj <> endobj Class: Euascomycetes The typical morphology of . 2015). were studied for the Gliocladium (with strongly convergent phialides) and Verticillium (with straight and moderately divergent phialides) are closely related genera. 2004; Anees et al. This refers to the ability of several Trichoderma species to form mutualistic endophytic relationships with several plant species. Three different families of repetitive sequences include: the 3540bp repetitive extragenic pallindromic (REP) sequence, the 124127bp enterobacterial repetitive intergenic consensus (ERIC) sequence and 154bp BOX (composed of the box A, B and C subunits) element. Correct identification will provide information on understanding the interparasitic relationship with target pathogens and the subsequent environmental fate of the antagonist needed for effective application. Peritonitis [319], pulmonary, perihepatic, and disseminated infections [1924] have so far been reported. Bacteria grow tremendously fast when supplied with an abundance of nutrients. As the conidia are formed, scattered blue-green or yellow-green patches become visible. % Genetic diversity, mating types and phylogenetic analysis of Indian races of, Krmczi P, Danilovi G, Manczinger L, Jovanovi L, Pankovi D, et al. 8600 Rockville Pike Pramod W. Ramteke, Email: moc.oohay@eketmarwp. Twenty isolates of Trichoderma were obtained from healthy tomato (Solanum lycopersicum cv. 0000002313 00000 n http://www.austin.cc.tx.us/microbugz/03morphology.html, accessed January 14, 2005. They usually turn into a different color, from the center outwards. There are several mechanisms involved in Trichoderma antagonism, namely, antibiosis whereby the antagonist fungus produces antibiotics, competes for nutrients and mycoparasitism, whereas Trichoderma directly attacks the plant pathogen by excreting lytic enzymes such as chitinases and -1,3 glucanases (Kubicek et al. and their application as a biocontrol agent in the field, it is important to understand their biodiversity. 0000007880 00000 n 4). BOX-PCR banding pattern showed a total of 200 fragments in the range of 2504000bp. Several species of Trichoderma have been used as biological control agents to manage diseases of vegetable and other crops (Solanki et al. Comparison of five rep-PCR genomic fingerprinting methods for differentiation of fecal, Ospina-Giraldo MD, Royse DJ, Thon MR, Chen X, Romaine CP. 0000098924 00000 n Image 11: A colony of yeast on the agar plate. official website and that any information you provide is encrypted Washington State University. 2013). Nickel contamination is a serious environmental issue that requires immediate action. 2011; Malmierca et al. However, it was ellipsoidal and obovoid in rest of the eight isolates (UNT13, UNT70, UNS28, UNS30, NAS46, ALS47 and NAT03). Tree was constructed by the neighbour-joining method. Thus, in present study, comparative nucleotide sequencing of EF1a gene was performed to distinguish and identify antagonistic Trichoderma isolates. Bacterial Colony Morphology: An Important Precursor. Cultural characteristics: Cultural characteristics comprising growth rate, colour and colony appearance were examined. Molecular and morphological characterization of green mold, de los Santos-Villalobos S, Guzmn-Ortiz DA, Gomez-Lim MA, Dlano-Frier JP, et al. For the molecular identification of ergosterol producing isolates, ERG1F (5-CGCTCCGTGCTTCTTCTTCTC-3) and EGR1R (5-CTTCTTCTCTCCCGTCTCC-3) primers were used. are common soil microorganisms that play an important role in limiting phytopathogenic microorganisms, improving plant growth and degrading plant biomass. The numbers given over branches indicate bootstrap coefficient. 0000019651 00000 n Trichoderma (Ascomycetes, Hypocreales) strains that have warted conidia are traditionally identified as T. viride, the type species of Trichoderma. FOIA Trichoderma harzianum Jn14 conidia, phialides, and colony. Consequently, more efficient Trichoderma isolates with high antagonistic potential capabilities are needed for successful biological control systems. The predominant specie was Trichoderma harzianum followed by Trichoderma viride. Ellipsoidal and sub-globose to globose conidia were noticed in thirteen isolates (UNT60, UNT64, UNT68, NAT69, NAT70, ALT73, DET89, DET94, HAT96, UNT38, UNS63, UNT09 and DET02). <>/XObject<>/ExtGState<>/Pattern<>/ProcSet[/PDF/Text/ImageB/ImageC/ImageI] >>/Annots[ 15 0 R] /MediaBox[ 0 0 612 792] /Contents 4 0 R/Group<>/Tabs/S/StructParents 0>> 0000274257 00000 n In fact there is a book called Bergey's Manual of Determinative Bacteriology (commonly termed Bergey's Manual) that describes the majority of bacterial species identified by scientists so far. 0000006630 00000 n However, their identity and clinical significance remain unconfirmed and doubtful [531]. Testing the Biological Control Ability of Trichoderma . However, the antifungal effects of Trichoderma and their metabolites on zoonotic fungal pathogens are largely unknown. Trichoderma stramenium; Colony diameter (PDA, 25C, 3 days) 90 in 2 days: 33-72: 36-49: 55-67: Colony color: . 2012; Al-Sadi et al. One approach involves the exploitation of ubiquitously conserved known genes that display sequence variation. However, two morphologically distinct types of conidial warts (I and II) have been found. These techniques are important not only for understanding their ecological role in the rhizosphere, but also to characterize the biological control agents for registration and patenting biocontrol strains, recognizing the strains, quality checking during production and ecological characterization (Plimmer 1993). Trichoderma is a genus of fungi in the family Hypocreaceae that is present in all soils, where they are the most prevalent culturable fungi. The PCR reactions were carried out in 25l reaction mixture containing 10 PCR buffer, 50ng DNA template, 2mM MgCl2, 0.25mM dNTP mixture and 0.25M each of primer, and one unit of Taq Polymerase (Bangalore Genie, India). For example, smooth, glistening, rough, dull (opposite of glistening), rugose (wrinkled), etc. 0000014973 00000 n http://biology.clc.uc.edu/fankhauser/Labs/Microbiology/Yeast_Plate_Count/07_yeast_0.2mL_plate_P7201181.jpg, accessed January 14, 2005. It does not have the typical structure of a bacterium, since it is not shaped like a coconut, a bacillus or a spiril. The present study evaluated whether the inoculation of Bacillus subtilis, Trichoderma harzianum, and rock powder into the soil would benefit soil fertility and plant growth in vase conditions. There was a positive correlation between the antagonistic capacity of Trichoderma isolates towards fungal pathogens and their lytic enzyme production. and transmitted securely. endobj For comparison of growth, colony appearance and morphological features, discs of fresh monosporic Trichoderma cultures were transferred to 9 cm Petri dishes containing 20 ml of either PDA, CMD (cornmeal dextrose agar) or SNA (synthetic low nutrient agar), which were cultured at 15, 20, 25, 30 and 35C, with 12 hour photoperiod. Recent evidences indicated the importance of the sterol biosynthetic pathway in inducing plant defense-related gene expression in both the antagonistic fungus and the plant (Cardoza et al. This powder was taken in a centrifuge tube and 2 CTAB (hexadecyltrimethyl ammonium bromide) buffer (15ml) was added in each tube separately. Trichoderma is a very common genus especially in soil and decaying wood. are the teleomorph of some Trichoderma species. The first (PCA1) and second (PCA2) principal components were accounted for 20.47 and 11.06%, respectively. The identification of Trichoderma species has been mainly determined by morphological features such as conidia and phialides growth rate, the colony color, size, and length. Their phylogenetic positions are determined by analyses of the combined partial sequences of translation elongation factor 1-alpha and the . On the . The first two principal component axis of the biplot accounted for 25.54% (PC1) and 27.36% (PC2) of the total variation of the pathogenantagonist interaction. 2015). Comparison of molecular and phenetic typing methods to assess diversity of selected members of the genus, Rai S, Kashyap PL, Kumar S, Srivastava AK, Ramteke PW. Based on these studies, we speculate that the species composition, distribution, and genetic structure of Trichoderma on the tomato rhizosphere may be different. These findings are in agreement with earlier studies, where no defined correlations between genetic variability assessed by random markers (e.g. The interaction of biological control agents versus four different fungal pathogens showed significant differences in growth inhibition of the pathogen isolates (Table1). Five primers viz., OPA-2 (TGCCGAGCTG), OPA-3 (AGTCAGCCAC), OPA-13 (CAGCACCCAC), OPA-15 (TTCCGAACCC) and OPA-18 (AGGTGACCGT) produced a total of 641 fragments among all the 20 isolates (Fig. 0000019461 00000 n Since, species of Trichoderma are reported as the causal agent of green mould disease (Ospina-Giraldo et al. Trichoderma infections are opportunistic [908] and develop in immunocompromised patients, such as neutropenic cases and transplant recipients, as well as patients with chronic renal failure, chronic lung disease, or amyloidosis. Briefly, for each fungal isolates, fresh mycelium (~5g) was dried on sterile blotter paper and was ground in liquid nitrogen to make a fine powder. This may be due to the ubiquitous nature and seemingly random chromosomal distribution of random repeats in Trichoderma genome, giving rise to simultaneous PCR amplification of multiple genomic regions (Rai et al. A lobate margin has rounded finger-like growths that spread outward. However, the results of these studies demonstrated that besides the natural ecosystems, the investigation of agricultural soils also reveals important information about Trichoderma biodiversity. 3). Meanwhile, deletion of vps8, vps10, and vps35 did not influenced the yeast colony morphology, and achieved higher Human urokinase-type plasminogen activator (uPA) secretion on fibrin-containing medium (Agaphonov et al. The PCR reaction was carried out in a 25-l reaction mixture containing the following: 10 PCR buffer, 50ng DNA template, 2mM MgCl2, 0.25mM dNTP mixture and 0.25M each of primer, and one unit of Taq Polymerase (Bangalore Genie, India). Accessibility Interestingly, the interaction of indigenous Trichoderma isolates with four different fungal pathogens resulted in significantly different amounts of pathogen inhibition. Further, combined studies including biological, biochemical and molecular technologies, are essential to select indigenous antagonistic Trichoderma isolates that can be used under different environmental conditions. Clade A comprised mainly T. harzianum (UNT60, UNT64, UNT68, NAT69 and UNT70), T. viride (UNT09, DET02 and NAT03), T. koningii (UNS63) and T. virens (UNS28) and showed very high homology to the nearest ERG1 sequence of H. lixii, T. arundinaceum and T. reesei submitted in NCBI GenBank. A perusal of data indicated that there was a significant difference in growth rate among isolates. Genetic variability of squalene epoxidase (ERG1) gene among these isolates in addition to their differences in aggressiveness toward multiple fungal pathogens suggest that combinations of isolates could further be applied in both greenhouse and field studies to obtain resistance against multiple fungal pathogens in tomato crop.
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