Morphomolecular Identification of Trichoderma sp. and their Mycoparasitic Activity Against Soil Borne Pathogens

This study was carried out to identify and characterize Trichoderma species isolated from rhizospheric soil of Uttar Pradesh, India, by using single spore technique. Morphological, cultural and molecular characterization were done with sequence analysis of the internal transcribed spacer (ITS) region. The classes were compared with morphological identification and rDNA sequence data for every class of all Trichoderma strains were of the same identity. These strains belonged to T. harzianum (Th azad), T. viride (01PP), T. asperellum (Tasp/CSAU), T. Koningii [TK (CSAU)], T. atroviride (71L), T. longibrachiatum (21PP), T. virens [Tvi (CSAU)], T. reesei [Tr (CSAU)], T. aggressivum [T.agg(CSAU)], T. aureoviride [T. avi (CSAU)], T. citrinoviride [T. cvi (CSAU)], T. erinaceum [T. eri (CSAU)], T. koningiopsis [T. kop (CSAU)], T. tomentosum [T. tos (CSAU)], T. mintisporum [T. mip (CSAU)], T. pubscenes [T. sce (CSAU)], T. saturnisporum [T. ssp (CSAU)], T. spirale [T. sp. (CSAU)]. Morphological studies were based on the colony appearance, growth rate and microscopic features such as branching patterns of conidiophores, the arrangement of phialospores and their shape, size and color. The 5.8S-ITS regions of the Trichoderma strains were amplified using ITS1 and ITS4 primers. The rRNA based analysis is a central method used not only to explore microbial diversity but also to identify new strains. Validations of ITS marker with 18 Trichoderma sp. were done and their sequences were deposited at NCBI GenBank their permanent accession no. were allotted.

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Morphological and molecular identification of ectomycorrhizal fungi associated with Persian oak (Quercus brantii Lindl.) tree

Journal of Forest Science ◽

10.17221/31/2020-jfs ◽

2020 ◽

Vol 66 (No. 6) ◽

pp. 244-251

Author(s):

Behnaz Yousefshahi ◽

Masoud Bazgir ◽

Samad Jamali ◽

Fatemeh Valizadeh Kakhki

Keyword(s):

Its Region ◽

Molecular Techniques ◽

Molecular Methods ◽

Morphological Identification ◽

Tree Roots ◽

Microscopic Features ◽

Zagros Zone ◽

Oak Tree ◽

Ecm Fungi ◽

Quercus Brantii

Identification of ectomycorrhizal (ECM) fungi in different ecosystems has major significance. In this research, to identity ECM fungi, we used two methods including the morphological method and the molecular method that is more precise. Basidiocarp collection of fungi associated with oak tree (Quercus brantii Lindl.) roots was carried out in the spring season 2016 and was identified by morphological and molecular methods. We also checked macroscopic and microscopic features and measured each structure using BioloMICS Measures software. To verify the morphological identification, the internal transcribed spacer (ITS) region was amplified by PCR using the primer pair ITS1/ITS4, and the sequences were analyzed. According to the morphological observations, the identified species were Amanita crocea, Boletus comptus, Tricholoma giganteum, and Inocybe rimosa. Besides, based on molecular techniques by comparing sequences, we identified five species out of the eight ones as A. crocea and other species as T. giganteum, I. rimosa and B. comptus. Both morphological and molecular methods are necessary for identifying ECM fungi associated with tree roots in the Zagros zone in the west of Iran.

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Molecular differentiation of the Murraya paniculata Complex (Rutaceae: Aurantioideae: Aurantieae)

BMC Evolutionary Biology ◽

10.1186/s12862-019-1555-4 ◽

2019 ◽

Vol 19 (1) ◽

Author(s):

Chung Huy Nguyen ◽

G. Andrew C. Beattie ◽

Anthony M. Haigh ◽

Inggit Puji Astuti ◽

David J. Mabberley ◽

...

Keyword(s):

Sequence Data ◽

Its Region ◽

Molecular Dating ◽

Putative Hybrid ◽

Citrus Industry ◽

Morphological Studies ◽

Murraya Paniculata ◽

Taxonomic Uncertainty ◽

Liberibacter Asiaticus ◽

Cryptic Taxa

Abstract Background Orange jasmine has a complex nomenclatural history and is now known as Murraya paniculata (L.) Jack. Our interest in this common ornamental stemmed from the need to resolve its identity and the identities of closely related taxa as hosts of the pathogen ‘Candidatus Liberibacter asiaticus’ and its vector Diaphorina citri. Understanding these microbe-vector-plant relationships has been hampered by taxonomic confusion surrounding Murraya at both the generic and specific levels. Results To resolve the taxonomic uncertainty, six regions of the maternally-inherited chloroplastal genome and part of the nuclear-encoded ITS region were amplified from 85 accessions of Murraya and Merrillia using the polymerase chain reaction (PCR). Clustering used maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI). Chronograms were produced for molecular dating, and to test the monophyly of Murraya rigorously, using selected accessions of Murraya and 26 accessions of the Rutaceae and Simarubaceae. Sequence data from the ITS and chloroplastal regions suggest that Murraya paniculata (sensu (Swingle WT and Reece CR, The Citrus Industry, p. 190–430, 1967)) can be separated into four distinct but morphologically somewhat cryptic taxa: Murraya paniculata (sensu (Mabberley DJ, Taxon 65:366–371, 2016)), M. elongata, M. sumatrana and M. lucida. In addition, Murraya omphalocarpa was identified as a putative hybrid of M. paniculata and M. lucida with two geographically isolated nothovarieties representing reciprocal crosses. Murraya is monophyletic, and molecular dating suggests that it diverged from Merrillia during the Miocene (23–5 Ma) with this Murraya group speciating and dispersing during the Middle Miocene onwards. Conclusions The accessions from Asia and Australasia used in this study grouped into biogeographical regions that match herbarium specimen records for the taxa that suggest natural allopatric distributions with limited overlap and hybridity. Murraya paniculata has been distributed around the world as an ornamental plant. The division of the Murraya paniculata complex into four species with a rare hybrid also confirms morphological studies.

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First Report of Brown Rot Caused by Monilinia fructicola on Nectarine in Serbia

Plant Disease ◽

10.1094/pdis-08-12-0718-pdn ◽

2013 ◽

Vol 97 (1) ◽

pp. 147-147 ◽

Cited By ~ 9

Author(s):

J. Hrustić ◽

M. Mihajlović ◽

B. Tanović ◽

G. Delibašić ◽

I. Stanković ◽

...

Keyword(s):

Prunus Persica ◽

Its Region ◽

Fruit Rot ◽

Economic Losses ◽

Morphological Identification ◽

Single Spore ◽

Daily Growth ◽

Causal Organism ◽

First Report ◽

Apple Fruits

In August 2011, nectarine (Prunus persica (L.) Batsch var. nucipersica (Suckow) C. K. Schneid) fruit originated from Oplenac region with symptoms of fruit rot was collected at a green market in Belgrade. Fruit had large, brown, sunken lesions covered with grayish brown tufts. Symptoms resembled those caused by species of Monilinia including M. laxa, M. fructigena, or M. fructicola (2). In order to isolate the causal organism, small superficial fragments of pericarp were superficially disinfected with commercial bleach and placed on potato dextrose agar (PDA). The majority (32 out of 33) isolates formed rosetted non-sporulating colonies with lobed margins resembling those of M. laxa. However, one isolate (Npgm) produced an abundant, grayish-white colony with even margins and concentric rings of sporogenous mycelium, resembling those described for M. fructicola (2). Conidia were one-celled, hyaline, ellipsoid to lemon shaped, 7.38 to 14.76 × 4.92 to 9.84 μm, and borne in branched monilioid chains. The average daily growth on PDA at 24°C was 10.9 mm. A single-spore isolate of Npgm was identified as M. fructicola based on the morphology of colony and conidia, temperature requirements, and growth rate (2). Morphological identification was confirmed by an amplified product of 535 bp using genomic DNA extracted from the mycelium of pure culture and species-specific PCR for the detection of M. fructicola (2). The ribosomal internal transcribed spacer (ITS) region of rDNA of Npgm was amplified and sequenced using primers ITS1/ITS4. Sequence analysis of ITS region revealed 100% nucleotide identity between the isolate Npgm (GenBank Accession No. JX127303) and 17 isolates of M. fructicola from different parts of the world, including four from Europe (FJ411109, FJ411110, GU967379, JN176564). Pathogenicity of the isolate Npgm was confirmed by inoculating five surface-disinfected mature nectarine and five apple fruits by placing a mycelial plug under the wounded skin of the fruit. Nectarine and apple fruits inoculated with sterile PDA plugs served as a negative controls. After a 3-day incubation at 22°C, inoculated sites developed brown lesions and the pathogen was succesfully reisolated. There were no symptoms on the control nectarine or apple fruits. M. fructicola is commonly present in Asia, North and South America, New Zealand, and Australia, while in the EPPO Region the pathogen is listed as an A2 quarantine organism (3). In Europe, the first discovery of M. fructicola was reported in France and since then, it has been found in Hungary, Switzerland, the Czech Republic, Spain, Slovenia, Italy, Austria, Poland, Romania, Germany, and Slovakia (1). Most recently, M. fructicola was found on stored apple fruits in Serbia (4). To our knowledge, this is the first report of M. fructicola decaying peach fruit in Serbia. These findings suggest that the pathogen is spreading on its principal host plants and causing substantial economic losses in the Serbian fruit production. References: (1) R. Baker et al. European Food Safety Authority. Online publication. www.efsa.europa.eu/efsajournal . EFSA J. 9:2119, 2011. (2) M. J. Côté. Plant Dis. 88:1219, 2004. (3) OEPP/EPPO. EPPO A2 list of pests recommended for regulation as quarantine pests. Version 2009-09. http://www.eppo.org/QUARANTINE/listA2.htm . (4). M. Vasic et al. Plant Dis. 96:456, 2012.

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Phylogenetic and morphological studies in Xylodon (Hymenochaetales, Basidiomycota) with the addition of four new species

MycoKeys ◽

10.3897/mycokeys.47.31130 ◽

2019 ◽

Vol 47 ◽

pp. 97-137 ◽

Cited By ~ 4

Author(s):

Janett Riebesehl ◽

Eugene Yurchenko ◽

Karen K. Nakasone ◽

Ewald Langer

Keyword(s):

New Species ◽

New Zealand ◽

Sequence Data ◽

Phylogenetic Analyses ◽

Its Region ◽

Morphological Data ◽

Similar Species ◽

New Combinations ◽

Line Drawings ◽

Morphological Studies

Xylodon (Hymenochaetales, Basidiomycota) is the largest segregate genus of Hyphodontia s.l. Based on molecular and morphological data, 77 species are accepted in Xylodon to date. Phylogenetic analyses of ITS and 28S sequences, including 38 new ITS and 20 28S sequences of Xylodon species, revealed four species new to science. The new taxa X.exilis, X.filicinus, X.follis and X.pseudolanatus from Taiwan, Nepal, Réunion, Belize, and USA are described and illustrated. In addition, species concepts for Odontiavesiculosa from New Zealand and Xylodonlanatus from U.S.A. are revised and the new name X.vesiculosus is proposed. Phylogenetic analyses of the ITS region placed X.spathulatus, X.bubalinus and X.chinensis in a strongly supported clade and demonstrated that they are conspecific. Palifer and Odontiopsis are synonymised under Xylodon based on morphological and sequence data. The following new combinations are proposed: X.erikssonii, X.gamundiae, X.hjortstamii, X.hyphodontinus, X.septocystidiatus and X.verecundus. Line drawings of X.cystidiatus, X.hyphodontinus, X.lanatus and X.vesiculosus, as well as photographs of X.raduloides basidiomata, are provided. A key to X.lanatus and similar species is presented.

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SINGLE SPORE ISOLATION AND MOLECULAR CHARACTERIZATION OF TRICHODERMA SPECIES FROM AGRICULTURAL FIELDS OF ZHEJIANG PROVINCE, CHINA

Pakistan Journal of Phytopathology ◽

10.33866/phytopathol.030.02.0464 ◽

2018 ◽

Vol 30 (2) ◽

pp. 115

Author(s):

Baharullah Khattak ◽

Hong K. Wang ◽

Fucheng Lin ◽

Saif Ullah ◽

Qaiser Jamal ◽

...

Keyword(s):

Plant Pathogens ◽

Agricultural Soils ◽

Biological Control Agent ◽

Control Agent ◽

Soil Samples ◽

Zhejiang Province ◽

Single Spore ◽

Trichoderma Spp ◽

Trichoderma Species ◽

Trichoderma Sp

It is well known that Trichoderma spp. can be used as a biological control agent against variety of plant pathogens. In this study, Trichoderma species were isolated from rhizosphere samples of various sites in Zhejiang province, China. A total of 41 soil samples were processed for the isolation of single cell culture of Trichoderma sp. Out of which, 32 species of the genus Trichoderma, were isolated and characterized by using single spore isolation method. The isolates were subjected to sequencing, for precise identification up to species level. Trichoderma harzianum was isolated from maximum (09) number of soil samples, followed by T. velutinum, isolated from 3 soil samples. The other isolated species of the genus Trichoderma were; lixii, atroviride, koningii, longibrachatum, rossicum, gamsii, koningiopsis, tomentosum, afroharzianum, viride, citroviride, aureoviride, simmonsii and hamatum. It was concluded that Trichoderma sp. were abundantly found in the agricultural soils of Zhejiang province, China.

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Schoenus (Cyperaceae) is not monophyletic based on ITS nrDNA sequence data

Australian Systematic Botany ◽

10.1071/sb15046 ◽

2016 ◽

Vol 29 (5) ◽

pp. 265 ◽

Cited By ~ 9

Author(s):

Paul M. Musili ◽

Adele K. Gibbs ◽

Karen L. Wilson ◽

Jeremy J. Bruhl

Keyword(s):

Bayesian Inference ◽

Internal Transcribed Spacer ◽

Sequence Data ◽

Its Region ◽

Bootstrap Support ◽

Nuclear Rdna ◽

Its Nrdna ◽

Rdna Sequence Data ◽

Strong Bootstrap Support ◽

Lines Of Evidence

We used nuclear rDNA-sequence data from the internal transcribed spacer (ITS) region to test the monophyly of Schoenus by using maximum parsimony and Bayesian inference. Schoenus is not monophyletic, with strong bootstrap support for most branches and congruence across analyses. nrITS does not resolve terminal taxa fully and, therefore, needs to be used in combination with other lines of evidence to address questions of species limits.

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Reconsideration of the ‘well-known’ hypotrichous ciliate Pleurotricha curdsi (Shi et al., 2002) Gupta et al., 2003 (Ciliophora, Sporadotrichida), with notes on its morphology, morphogenesis and molecular phylogeny

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.000377 ◽

2015 ◽

Vol 65 (Pt_9) ◽

pp. 3216-3225 ◽

Cited By ~ 7

Author(s):

Xiaoteng Lu ◽

Chen Shao ◽

Yuhe Yu ◽

Alan Warren ◽

Jie Huang

Keyword(s):

Sequence Data ◽

Southern China ◽

Phylogenetic Analyses ◽

Large Body ◽

Small Subunit ◽

Variable Number ◽

Ssu Rdna ◽

Intermediate Form ◽

Rdna Sequence Data

The oxytrichid species Pleurotricha curdsi (Shi et al., 2002) Gupta et al., 2003, isolated from a tributary of the Yangtze River in the Mudong district of Chongqing, southern China, was reinvestigated with emphasis on its morphology, morphogenesis and small-subunit (SSU) rDNA-based phylogeny. Compared with three previously described populations, the Mudong population of P. curdsi is characterized by its large body size, 170–295 × 65–110 μm in vivo, and by having a variable number of right marginal rows, either two or three. Likewise, the number of right marginal rows anlagen (RMA) is also variable, i.e. usually two, but sometimes several small extra anlagen that give rise to the formation of the third row, are present to the left of the RMAs. We posit that the Mudong population is an intermediate form between the three previously described populations. Phylogenetic analyses based on the SSU rDNA sequence data show that all populations of P. curdsi cluster with the type species of the genus, Pleurotricha lanceolata, in a clade nested within the Oxytrichidae.

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Higher-level relationships among the eucalypts are resolved by ITS-sequence data

Australian Systematic Botany ◽

10.1071/sb00039 ◽

2002 ◽

Vol 15 (1) ◽

pp. 49 ◽

Cited By ~ 82

Author(s):

Dorothy A. Steane ◽

Dean Nicolle ◽

Gay E. McKinnon ◽

René E. Vaillancourt ◽

Brad M. Potts

Keyword(s):

Internal Transcribed Spacer ◽

Sequence Data ◽

Its Region ◽

Molecular Data ◽

Nuclear Ribosomal Dna ◽

Its Sequence ◽

Phylogenetic Groups ◽

Single Section ◽

Western Australian ◽

Genus Eucalyptus

This expanded survey of ITS sequences represents the largest analysis of molecular data ever attempted on Eucalyptus. Sequences of the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA were included in an analysis of 90 species of Eucalyptus s.s. and 28 species representing eight other genera (Allosyncarpia, Angophora, Arillastrum, Corymbia, Eucalyptopsis, Stockwellia, Lophostemon and Metrosideros). The results of the study indicate that Angophora and Corymbia form a well-supported clade that is highly differentiated from Eucalyptus s.s. Corymbia species are divided between two clades, one of which may be the sister to Angophora. Allosyncarpia, Arillastrum, Eucalyptopsis and ‘Stockwellia’ are also highly differentiated from Eucalyptus s.s. If the genus Eucalyptus is to be expanded to include Angophora and Corymbia(sensu Brooker 2000), ITS data suggest that Allosyncarpia, Eucalyptopsis, ‘Stockwellia’ and potentially Arillastrum should also be included in Eucalyptus s.l. The ITS data suggest that subg. Symphyomyrtus is paraphyletic and that subg. Minutifructus should be included within it. Within subg.Symphyomyrtus, only sect. Maidenaria appears to be monophyletic. Sections Adnataria and Dumaria are probably monophyletic; sections Exsertaria and Latoangulatae are very close and probably should be combined in a single section. Section Bisectae is polyphyletic and is divided into two distinct lineages. The phylogenetic groups depicted by ITS data are consistent with the frequency of natural inter-specific hybridisations as well as data from controlled crosses within subgenus Symphyomyrtus. The ITS data illustrate that subg. Idiogenes and western Australian monocalypts are early evolutionary lines relative to E. diversifolia, E. rubiginosa (monotypic subg. Primitiva) and the eastern monocalypts and that subg. Primitiva should be sunk into subg. Eucalyptus. Subgenus Eudesmia may be monophyletic, grouping with subgenera Idiogenes and Eucalyptus. Further work is required to confirm the phylogenetic positions of the monotypic subgenera Alveolata, Cruciformes, Acerosae and Cuboidea.

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Biological and Phylogenetic Analyses Suggest that Two Cryphonectria spp. Cause Cankers of Eucalyptus in Africa

Plant Disease ◽

10.1094/pdis.2003.87.11.1329 ◽

2003 ◽

Vol 87 (11) ◽

pp. 1329-1332 ◽

Cited By ~ 18

Author(s):

J. Roux ◽

H. Myburg ◽

B. D. Wingfield ◽

M. J. Wingfield

Keyword(s):

South Africa ◽

South America ◽

South African ◽

Sequence Data ◽

Phylogenetic Analyses ◽

Eucalyptus Grandis ◽

Its Region ◽

South African Isolate ◽

Dna Sequence Data ◽

The Republic

Cryphonectria cubensis is an economically important pathogen of commercial Eucalyptus spp. Differences have been reported for disease symptoms associated with Cryphonectria canker in South Africa and other parts of the world, and recent DNA-based comparisons have confirmed that the fungus in South Africa is different from that in South America and Australasia. During a disease survey in the Republic of Congo, Cryphonectria canker was identified as an important disease on Eucalyptus grandis and E. urophylla. In this study, we compared Congolese and South African isolates of C. cubensis using DNA sequence data and pathogenicity under greenhouse conditions. The β-tubulin and internal transcribed spacer (ITS) region sequences show that C. cubensis in Congo is different from the fungus in South Africa and that Congolese isolates group most closely with South American isolates. Furthermore, pathogenicity tests showed that a South African isolate was more aggressive than two Congolese isolates. We conclude that two distinct Cryphonectria spp. occur in Africa and hypothesize that the fungus in the Congo probably was introduced into Africa from South America. Both fungi are important pathogens causing disease and death of economically important plantation trees. However, they apparently have different origins and must be treated separately in terms of disease management and quarantine considerations.

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Revealing the endophytic mycoflora in tea (Camellia sinensis) leaves in Sri Lanka: the first comprehensive study

Phytotaxa ◽

10.11646/phytotaxa.514.3.5 ◽

2021 ◽

Vol 514 (3) ◽

pp. 247-260

Author(s):

KASUN THAMBUGALA ◽

DINUSHANI DARANAGAMA ◽

SAGARIKA KANNANGARA ◽

THENUKA KODITUWAKKU

Keyword(s):

Sri Lanka ◽

Camellia Sinensis ◽

Endophytic Fungi ◽

Sequence Data ◽

Fungal Endophytes ◽

Fungal Endophyte ◽

Tea Leaves ◽

Molecular Sequence Data ◽

Molecular Sequence ◽

Rdna Sequence Data

Endophytic fungi are a diverse group of microorganisms that live asymptomatically in healthy tissues of host and they have been reported from all kinds of plant tissues such as leaves, stems, roots, flowers, and fruits. In this study, fungal endophytes associated with tea leaves (Camellia sinensis) were collected from Kandy, Kegalle, and Nuwara Eliya districts in Sri Lanka and were isolated, characterized, and identified. A total of twenty endophytic fungal isolates belonging to five genera were recovered and ITS-rDNA sequence data were used to identify them. All isolated endophytic fungal strains belong to the phylum Ascomycota and the majority of these isolates were identified as Colletotrichum species. Phyllosticta capitalensis was the most commonly found fungal endophyte in tea leaves and was recorded in all three districts where the samples were collected. This is the very first investigation on fungal endophytes associated with C. sinensis in Sri Lanka based on molecular sequence data. In addition, a comprehensive account of known endophytic fungi reported worldwide on Camellia sinensis is provided.

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