scholarly journals First Report of Mango Malformation Disease Caused by Fusarium pseudocircinatum in Mexico

Plant Disease ◽  
2014 ◽  
Vol 98 (11) ◽  
pp. 1583-1583 ◽  
Author(s):  
S. Freeman ◽  
G. Otero-Colina ◽  
G. Rodríguez-Alvarado ◽  
S. Fernández-Pavía ◽  
M. Maymon ◽  
...  
Keyword(s):  
South Africa ◽  
Phylogenetic Analyses ◽  
Mangifera Indica ◽  
Large Subunit ◽  
Elongation Factor ◽  
Economic Loss ◽  
Causal Agent ◽  
First Report ◽  
The Usa ◽  
Outdoor Nursery

Mango (Mangifera indica L.) malformation disease (MMD) is one of the most important diseases affecting this crop worldwide, causing severe economic loss due to reduction of yield. After the first report in India in 1891 (3), MMD has spread worldwide to most mango-growing regions. Several species of Fusarium cause the disease, including F. mangiferae in India, Israel, the USA (Florida), Egypt, South Africa, Oman, and elsewhere; F. sterilihyphosum in South Africa and Brazil; F. proliferatum in China; F. mexicanum in Mexico; and recently, F. tupiense in Brazil (1,2,3,4). Besides F. mexicanum, F. pseudocircinatum, not yet reported as a causal agent of MMD, was isolated in Mexico from affected inflorescences and vegetative malformed tissues (4). Symptoms of vegetative malformation caused by F. pseudocircinatum included hypertrophied, tightly bunched young shoots, with swollen apical and lateral buds producing misshapen terminals with shortened internodes and dwarfed leaves. Infected inflorescences of primary or secondary axes on affected panicles were shortened, thickened, and highly branched, while the peduncles became thick, remained green and fleshy, and branches profusely resembled a cauliflower in shape and size (3). Ten isolates of F. pseudocircinatum were recovered from cultivars Ataulfo, Criollo, Haden, and Tommy Atkins in Guerrero, Campeche, and Chiapas states and characterized. Isolates produced mostly 0-septate but occasionally 1- to 3-septate oval, obovoid, or elliptical aerial conidia (0-septate: 4 to 19 [avg. 8.7] × 1.5 to 4 [avg. 2.6] μm) in false heads in the dark and in short false chains under black light, unbranched or sympodially branched prostrate aerial conidiophores producing mono- and polyphialides, and sporodochia with straight or falcate conidia that were mostly 3- to 5-septate, but sometimes up to 7-septate (3-septate: 25 to 58 [avg. 41] × 2 to 3.3 [avg. 2.9] μm; 5-septate: 33.5 to 76.5 [avg. 56.7] × 2.5 to 6 [avg. 3.5] μm). Circinate sterile hyphae were rarely formed. Two representative isolates, NRRL 53570 and 53573, were subjected to multilocus molecular phylogenetic analyses of portions of five genes: nuclear large subunit 28S ribosomal RNA, β-tubulin, calmodulin, histone H3, and translation elongation factor (TEF)-1α (GenBank GU737456, GU737457, GU737290, GU737291, GU737371, GU737372, GU737425, GU737426, GU737398, and GU737399). Two pathogenicity tests were conducted with NRRL 53570 and 53573 on healthy 2-year-old nucellar seedlings of polyembryonic Criollo; 20 μl conidial suspensions (5 × 106 conidia/ml) of each isolate and water controls were inoculated separately on 15 buds on 3 different trees, as described previously (1). The following conditions were used in experiment 1: 24 to 27°C with light intensity of 16.2 to 19.8 •Mol m−2s−1 in the range of 400 to 700 nm, and photoperiods of 14 h light and 10 h dark. Typical vegetative disease symptoms were discernible in plants inoculated with NRRL 53570 (20%) and 53573 (7%) after 8 months. In experiment 2, after 3 months growth under the above conditions, seedlings were transferred to an outdoor nursery in Iguala, Guerrero. Typical vegetative symptoms of MMD were observed in 86.7 and 13.3% of the buds inoculated with F. pseudocircinatum NRRL 53570 and 53573, respectively, after 9 months. Isolates from typical symptomatic vegetative buds were confirmed as F. pseudocircinatum by sequencing a portion of their TEF-1α gene, thus fulfilling Koch's postulates. This is the first report of F. pseudocircinatum as a causal agent of MMD. References: (1) S. Freeman et al. Phytopathology 89:456, 1999. (2) C. S. Lima et al. Mycologia 104:1408, 2012. (3) W. F. O. Marasas et al. Phytopathology 96:667, 2006. (4) G. Otero-Colina et al. Phytopathology 100:1176, 2010.

scholarly journals Multilocus phylogenies reveal three new truffle-like taxa and the traces of interspecific hybridization in Octaviania (Boletaceae, Boletales)

IMA Fungus ◽  
2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Takamichi Orihara ◽  
Rosanne Healy ◽  
Adriana Corrales ◽  
Matthew E. Smith
Keyword(s):  
Rna Polymerase Ii ◽  
Phylogenetic Analyses ◽  
Large Subunit ◽  
Gene Tree ◽  
Phylogenetic Network ◽  
Elongation Factor ◽  
Morphological Observation ◽  
Tree Comparison ◽  
Unknown Species ◽  
The Usa

ABSTRACTAmong many convergently evolved sequestrate fungal genera in Boletaceae (Boletales, Basidiomycota), the genus Octaviania is the most diverse. We recently collected many specimens of Octaviania subg. Octaviania, including several undescribed taxa, from Japan and the Americas. Here we describe two new species in subgenus Octaviania, O. tenuipes and O. tomentosa, from temperate to subtropical evergreen Fagaceae forests in Japan based on morphological observation and robust multilocus phylogenetic analyses (nrDNA ITS and partial large subunit [LSU], translation elongation factor 1-α gene [TEF1] and the largest subunit of RNA polymerase II gene [RPB1]). Based on specimens from the Americas as well as studies of the holotype, we also taxonomically re-evaluate O. asterosperma var. potteri. Our analysis suggests that O. asterosperma var. potteri is a distinct taxon within the subgenus Octaviania so we recognize this as O. potteri stat. nov. We unexpectedly collected O. potteri specimens from geographically widespread sites in the USA, Japan and Colombia. This is the first verified report of Octaviania from the South American continent. Our molecular analyses also revealed that the RPB1 sequence of one O. tenuipes specimen was identical to that of a closely related species, O. japonimontana, and that one O. potteri specimen from Minnesota had an RPB1 sequence of an unknown species of O. subg. Octaviania. Additionally, one O. japonimontana specimen had an unusually divergent TEF1 sequence. Gene-tree comparison and phylogenetic network analysis of the multilocus dataset suggest that these heterogenous sequences are most likely the result of previous inter- and intra-specific hybridization. We hypothesize that frequent hybridization events in Octaviania may have promoted the high genetic and species diversity found within the genus.

scholarly journals ­­­Multi-locus phylogenies reveal three new truffle-like taxa and the traces of interspecific hybridization in Octaviania (Boletales)

2020 ◽  
Author(s):  
Takamichi Orihara ◽  
Rosanne Healy ◽  
Adriana Corrales ◽  
Matthew E. Smith
Keyword(s):  
Rna Polymerase Ii ◽  
Phylogenetic Analyses ◽  
Large Subunit ◽  
Gene Tree ◽  
Phylogenetic Network ◽  
Elongation Factor ◽  
Morphological Observation ◽  
Tree Comparison ◽  
Unknown Species ◽  
The Usa

Abstract Among many convergently evolved sequestrate fungal genera in Boletaceae (Boletales, Basidiomycota), the genus Octaviania is the most diverse. We recently collected many specimens of Octaviania subgenus Octaviania, including several undescribed taxa, from Japan and the Americas. Here we describe two new species in subgenus Octaviania, O. tenuipes and O. tomentosa, from temperate to subtropical evergreen Fagaceae forests in Japan based on morphological observation and robust multi-locus phylogenetic analyses (nrDNA ITS and partial large subunit [LSU], translation elongation factor 1-α gene [TEF1] and the largest subunit of RNA polymerase II gene [RPB1]). Based on specimens from the Americas as well as studies of the holotype, we also taxonomically re-evaluate the taxon O. asterosperma var. potteri. Our analysis suggests that O. asterosperma var. potteri is a distinct taxon within the subgenus Octaviania so we recognize this as O. potteri. We unexpectedly collected O. potteri specimens from geographically widespread sites in the USA, Japan and Colombia. This is the first verified report of Octaviania from the South American continent. Our molecular analyses also revealed that the RPB1 sequence of one O. tenuipes specimen was identical to that of a closely related species, O. japonimontana, and that one O. potteri specimen from Minnesota had an RPB1 sequence of an unknown species of O. subg. Octaviania. Additionally, one O. japonimontana specimen had an unusually divergent TEF1 sequence. Gene-tree comparison and phylogenetic network analysis of the multi-locus dataset suggest that these heterogenous sequences are most likely the result of previous inter- and intra-specific hybridization. We hypothesize that frequent hybridization events in Octaviania may have promoted the high genetic and species diversity within the genus.

scholarly journals Identification of Lasiodiplodia pseudotheobromae Causing Fruit Rot of Citrus in China

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 202
Author(s):  
Jianghua Chen ◽  
Zihang Zhu ◽  
Yanping Fu ◽  
Jiasen Cheng ◽  
Jiatao Xie ◽  
...  
Keyword(s):  
Phylogenetic Analyses ◽  
Elongation Factor ◽  
Morphological Characteristics ◽  
Citrus Fruit ◽  
Economic Loss ◽  
Fruit Rot ◽  
Post Inoculation ◽  
Citrus Reticulata ◽  
Postharvest Diseases ◽  
Citrus Reticulata Blanco

Considering the huge economic loss caused by postharvest diseases, the identification and prevention of citrus postharvest diseases is vital to the citrus industry. In 2018, 16 decayed citrus fruit from four citrus varieties—Satsuma mandarin (Citrus unshiu), Ponkan (Citrus reticulata Blanco cv. Ponkan), Nanfeng mandarin (Citrus reticulata cv. nanfengmiju), and Sugar orange (Citrus reticulata Blanco)—showing soft rot and sogginess on their surfaces and covered with white mycelia were collected from storage rooms in seven provinces. The pathogens were isolated and the pathogenicity of the isolates was tested. The fungal strains were identified as Lasiodiplodia pseudotheobromae based on their morphological characteristics and phylogenetic analyses using the internal transcribed spacer regions (ITS), translation elongation factor 1-α gene (TEF), and beta-tubulin (TUB) gene sequences. The strains could infect wounded citrus fruit and cause decay within two days post inoculation, but could not infect unwounded fruit. To our knowledge, this is the first report of citrus fruit decay caused by L. pseudotheobromae in China.

Affinities of the Boletus chromapes group to Royoungia and the description of two new genera, Harrya and Australopilus

2012 ◽  
Vol 25 (6) ◽  
pp. 418 ◽  
Author(s):  
Roy E. Halling ◽  
Mitchell Nuhn ◽  
Todd Osmundson ◽  
Nigel Fechner ◽  
James M. Trappe ◽  
...  
Keyword(s):  
New Genus ◽  
Phylogenetic Analyses ◽  
Large Subunit ◽  
Elongation Factor ◽  
Sister Group ◽  
Molecular Data ◽  
Translation Elongation ◽  
Spore Deposit ◽  
Large Subunit Rdna ◽  
Translation Elongation Factor 1Α

Harrya is described as a new genus of Boletaceae to accommodate Boletus chromapes, a pink-capped bolete with a finely scabrous stipe adorned with pink scabers, a chrome yellow base and a reddish-brown spore deposit. Phylogenetic analyses of large-subunit rDNA and translation elongation factor 1α confirmed Harrya as a unique generic lineage with two species, one of which is newly described (H. atriceps). Some Chinese taxa were recently placed in a separate genus, Zangia, supported by both morphology and molecular data. Multiple accessions from Queensland, Australia, support the synonymy of at least three species in a separate Australian clade in the new genus, Australopilus. The truffle-like Royoungia is also supported as a separate lineage in this clade of boletes. Even though it lacks stipe characters, it possesses the deep, bright yellow to orange pigments in the peridium. Additional collections from Zambia and Thailand represent independent lineages of uncertain phylogenetic placement in the Chromapes complex, but sampling is insufficient for formal description of new species. Specimens from Java referable to Tylopilus pernanus appear to be a sister group of the Harrya lineage.

Generic classification of some more hyphomycetes with solitary conidia borne on phialides

1998 ◽  
Vol 76 (9) ◽  
pp. 1570-1583 ◽  
Author(s):  
W Gams ◽  
K O'Donnell ◽  
H -J Schroers ◽  
M Christensen
Keyword(s):  
New Species ◽  
Phylogenetic Analyses ◽  
Large Subunit ◽  
Elongation Factor ◽  
28S Rdna ◽  
New Combination ◽  
Rdna Sequences ◽  
28S Rdna Sequences ◽  
Three New Species ◽  
Morphological Species

Unlike most phialide-producing fungi that liberate a multiplicity of conidia from each conidiogenous cell, only single conidia are formed on phialide-like conidiogenous cells in Aphanocladium, Verticimonosporium, and some species of Sibirina. A group of isolates obtained from soil of native Artemisia tridentata (sagebrush) grassland in Wyoming and from desert soil in Iraq is compared with these genera and classified as a fourth genus, Stanjemonium, honouring Stanley J. Hughes. Phylogenetic analyses of partial nuclear small- (18S) and large-subunit (28S) rDNA sequences indicate that Stanjemonium spp. form a monophyletic group with Emericellopsis. Sequences from the nuclear 18S and 28S rDNA were too conserved to resolve morphological species of Stanjemonium; however, phylogenetic analysis of b-tubulin and translation elongation factor 1a gene exons and introns resolved all species distinguished morphologically. Numerous conidiogenous cells or denticles are scattered along the cells of aerial hyphae in Aphanocladium and Stanjemonium spp., very rapidly collapsing into denticles in the former, somewhat more persistent and leaving broad scars in the latter. In Cladobotryum-Sibirina and Verticimonosporium spp., conidiogenous cells are discrete in terminal and intercalary whorls; phialides of the latter taxon are particularly swollen. The taxonomy of Aphanocladium is not yet resolved. Two species are recognized in Verticimonosporium. Three new species of Stanjemonium are described, and one new combination from Aphanocladium is proposed, along with one new species of Cladobotryum.Key words: Aphanocladium, Cladobotryum, conidiogenesis, hyphomycetes, molecular phylogeny, phialide, Stanjemonium, systematics, Verticimonosporium.

scholarly journals First Report of Leaf Rust of Blueberry Caused by Thekopsora minima on Vaccinium corymbosum in the Western Cape, South Africa

Plant Disease ◽  
2010 ◽  
Vol 94 (4) ◽  
pp. 478-478 ◽  
Author(s):  
L. Mostert ◽  
W. Bester ◽  
T. Jensen ◽  
S. Coertze ◽  
A. van Hoorn ◽  
...  
Keyword(s):  
South Africa ◽  
South African ◽  
Water Solution ◽  
Phylogenetic Analyses ◽  
The United States ◽  
Vaccinium Corymbosum ◽  
Tween 20 ◽  
Western Cape ◽  
The South ◽  
First Report

Southern highbush blueberry plants (Vaccinium corymbosum interspecific hybrids) showing rust-like symptoms were observed in July 2006 in Porterville in the Western Cape (WC), South Africa. Diseased plants were also found in Villiersdorp and George in the WC in 2007. In 2008, symptoms were observed in George, and in 2009, in all the previous reported areas. Cvs. Bluecrisp, Emerald, Jewel, Sharpblue, and Star were infected. Reddish-to-brown spots appeared on the adaxial surface of leaves and developed into yellow-to-orange erumpent uredinia with pulverulent urediniospores. Uredinia were hypophyllous, dome shaped, 113 to 750 μm wide, and occasionally coalescing. Urediniospores were broadly obovate, sometimes ellipsoidal or pyriform, with yellowish orange content, and measured 19 to 27 × 12 to 20 μm (average 24 × 15 μm, n = 30). Spore walls were echinulate, hyaline, 1 to 1.5 μm thick, and with obscure germ pores. No telia or teliospores were observed. Voucher specimens were lodged in the South African National Fungus Collection in Pretoria (PREM 60245). The isolate was initially identified as Thekopsora minima P. Syd. & Syd., based primarily on the absence of conspicuous ostiolar cells characteristic of Naohidemyces spp. (3). Genomic DNA was extracted from urediniospores. Approximately 1,400 bp were amplified spanning the 5.8S, ITS2, and 28S large subunit of the ribosomal DNA (1). The sequence (GU355675) shared 96% (907 of 942 bp; GenBank AF522180) and 94% (1,014 of 1,047 bp; GenBank DQ354563) similarities in the 28S portion, respectively, to those of Naohidemyces vaccinii (Wint.) Sato, Katsuya et Y. Hiratsuka and Pucciniastrum geoppertianum (Kuehn) Kleb, two of the three known rust species of blueberry (2). Although no sequences of T. minima were available for direct comparison, phylogenetic analyses of the 28S region strongly supported the South African blueberry rust as congeneric with T. guttata (J. Schröt.) P. Syd. & Syd. (GenBank AF426231) and T. symphyti (Bubák) Berndt (GenBank AF26230) (data not shown). Four 6-month-old cv. Sharpblue plants were inoculated with a suspension (approximate final concentration of 1 × 105 spores per ml) of fresh urediniospores in a water solution with 0.05% Tween 20. After incubation at 20°C for 48 h under continuous fluorescent lighting, the plants were grown in a glasshouse (18/25°C night/day temperatures). Identical uredinia and symptoms developed approximately 3 weeks after inoculation on the inoculated plants, but not on two control plants of cv. Sharpblue sprayed with distilled water and kept at the same conditions. The alternate host hemlock (Tsuga spp.) is not endemic to South Africa and not sold as an ornamental plant according to a large conifer nursery. Hosts of T. minima include Gaylussacia baccata, G. frondosa, Lyonia neziki, Menziesia pilosa, Rhododendron canadense, R. canescens, R. lutescens R. ponticum, R. prunifolium, R. viscosum, V. angustifolium var. laevifolium, V. corumbosum, and V. erythrocarpon (3). Visual inspection of possible hosts in the gardens in close proximity of Vaccinium production areas did not show any rust symptoms. To our knowledge, this is the first report of T. minima on blueberries outside of Asia and the United States (2). References: (1) M. C. Aime. Mycoscience 47:112, 2006. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Botany and Mycology Laboratory. Online publication. USDA-ARS, 2009. (3) S. Sato et al. Trans. Mycol. Soc. Jpn. 34:47, 1993.

Spegazzinia camelliae sp. nov. (Didymosphaeriaceae, Pleosprales), a new endophytic fungus from northern Thailand

Phytotaxa ◽  
2021 ◽  
Vol 483 (2) ◽  
pp. 117-128
Author(s):  
NAKARIN SUWANNARACH ◽  
JATURONG KUMLA ◽  
SAISAMORN LUMYONG
Keyword(s):  
Phylogenetic Analyses ◽  
Large Subunit ◽  
Elongation Factor ◽  
Small Subunit ◽  
Internal Transcribed Spacers ◽  
Nuclear Ribosomal Dna ◽  
Full Description ◽  
Translation Elongation ◽  
Spine Length ◽  
Elongation Factor 1 Alpha

A new endophytic ascomycete, described herein as Spegazzinia camelliae, was isolated from leaves of Camellia sinensis var. assamica collected from Nan Province, Thailand. This species is characterized by basauxic conidiophores and dark brown to blackish brown α and β conidia. It can be distinguished from previously described Spegazzinia species by the spine length of the α conidia and the size of the β conidia. Multi-gene phylogenetic analyses of the small subunit (SSU), large subunit (LSU) and internal transcribed spacers (ITS) of the nuclear ribosomal DNA (rDNA) and the translation elongation factor 1-alpha (tef1) genes also support S. camelliae is a distinct new species within Spegazzinia. A full description, color photographs, illustrations and a phylogenetic tree showing the position of S. camelliae are provided.

scholarly journals Multi-gene phylogeny and taxonomy of Amauroderma s. lat. (Ganodermataceae)

2020 ◽  
Vol 44 (1) ◽  
pp. 206-239 ◽  
Author(s):  
Y.-F. Sun ◽  
D.H. Costa-Rezende ◽  
J.-H. Xing ◽  
J.-L. Zhou ◽  
B. Zhang ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Phylogenetic Analyses ◽  
Large Subunit ◽  
Elongation Factor ◽  
Morphological Examination ◽  
Translation Elongation ◽  
Multiple Loci ◽  
Phylogenetic Studies ◽  
Molecular Phylogenetic

Amauroderma s.lat. has been defined mainly by the morphological features of non-truncate and double-walled basidiospores with a distinctly ornamented endospore wall. In this work, taxonomic and phylogenetic studies on species of Amauroderma s.lat. are carried out by morphological examination together with ultrastructural observations, and molecular phylogenetic analyses of multiple loci including the internal transcribed spacer regions (ITS), the large subunit of nuclear ribosomal RNA gene (nLSU), the largest subunit of RNA polymerase II (RPB1) and the second largest subunit of RNA polymerase II (RPB2), the translation elongation factor 1-α gene (TEF) and the β-tubulin gene (TUB). The results demonstrate that species of Ganodermataceae formed ten clades. Species previously placed in Amauroderma s.lat. are divided into four clades: Amauroderma s.str., Foraminispora, Furtadoa and a new genus Sanguinoderma. The classification of Amauroderma s. lat. is thus revised, six new species are described and illustrated, and eight new combinations are proposed. SEM micrographs of basidiospores of Foraminispora and Sanguinoderma are provided, and the importance of SEM in delimitation of taxa in this study is briefly discussed. Keys to species of Amauroderma s.str., Foraminispora, Furtadoa, and Sanguinoderma are also provided.

scholarly journals Molecular characterization and phylogenetic analyses of Lophodermella needle pathogens (Rhytismataceae) on Pinus species in the USA and Europe

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11435
Author(s):  
Jessa P. Ata ◽  
Kelly S. Burns ◽  
Suzanne Marchetti ◽  
Isabel A. Munck ◽  
Ludwig Beenken ◽  
...  
Keyword(s):  
Phylogenetic Analyses ◽  
Ribosomal Subunit ◽  
Elongation Factor ◽  
Translation Elongation ◽  
Phenotypic Characters ◽  
Elongation Factor 1 Alpha ◽  
The Usa ◽  
Phylogenetic Resolution ◽  
Elongation Factor 1

Increasing prevalence of conifer needle pathogens globally have prompted further studies on pathogen identification and a better understanding of phylogenetic relationships among needle pathogens. Several Lophodermella species can be aggressive pathogens causing needle cast in natural pine forests in the USA and Europe. However, their relationships with other Rhytismataceae species have historically been based on similarities of only limited phenotypic characters. Currently, no molecular studies have been completed to elucidate their relationships with other Lophodermella needle pathogens. This study collected and sequenced three gene loci, namely: internal transcribed spacer, large ribosomal subunit, and translation elongation factor 1-alpha, from five Lophodermella needle pathogens from North America (L. arcuata, L. concolor, L. montivaga) and Europe (L. conjuncta and L. sulcigena) to distinguish phylogeny within Rhytismatacaeae, including Lophophacidium dooksii. Phylogenetic analyses of the three loci revealed that all but L. conjuncta that were sampled in this study consistently clustered in a well-supported clade within Rhytismataceae. The multi-gene phylogeny also confirmed consistent nesting of L. dooksii, a needle pathogen of Pinus strobus, within the clade. Potential synapomorphic characters such as ascomata position and ascospore shape for the distinct clade were also explored. Further, a rhytismataceous species on P. flexilis that was morphologically identified as L. arcuata was found to be unique based on the sequences at the three loci. This study suggests a potential wider range of host species within the genus and the need for genetic characterization of other Lophodermella and Lophophacidium species to provide a higher phylogenetic resolution.

scholarly journals First Report of Postharvest Stem End Rot of mango fruit (Mangifera indica) caused by Lasiodiplodia theobromae in China.

Plant Disease ◽  
2021 ◽  
Author(s):  
Yanling Ma ◽  
Tanvir Ahmad ◽  
Yongquan Zheng ◽  
Nie Chengrong ◽  
Yang Liu
Keyword(s):  
Mangifera Indica ◽  
Causal Agent ◽  
Post Inoculation ◽  
Mango Fruit ◽  
Isolation Technique ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Fruit Samples ◽  
Fruit Surface ◽  
Β Tubulin

China is the second largest producer of mango in the world, a fruit has high nutritive value and a rich source of fiber (Kuhn et al., 2017). In late June 2019, a postharvest stem-end rot disease was observed in different local fruit markets (39°48'42.1"N 116°20'17.0"E) of the Fengtai district of Beijing, China. Black rot symptomatic lesions were observed on the fruit surface which initially started from the stem end of the mango fruit (Fig. 1). Approximately 45 % of mango fruits were affected with the disease. Symptomatic portions from collected fruit samples (n=40) were cut into small pieces (2mm2), rinsed with 1% NaClO for 20s and then washed three times with sterilized distilled water (SDW) for surface disinfection. The disinfected pieces were then placed on sterilized filter paper for drying. Later, these pieces were placed on Potato Dextrose Agar (PDA) plates and incubated at 28°C for seven days. The resulting fungal colonies were purified by the single spore isolation technique. The isolated fungal colonies were initially greenish to gray in color, later turning olive-black to black. Conidia were dark brown in color, oval-shaped, two-celled and measured 22.4 to 25.7 (24.06 ± 0.15) μm in length and 10.2 to 12.8 (11.3 ± 0.13) μm in width (n=36). Based on the symptoms, culture morphology and microscopic characters, Lasiodiplodia theobromae was suspected as the causal agent, and similar results were reported by Pavlic et al., 2004 and Burgess et al., 2006. For molecular identification, a multi-locus sequence analysis approach was used. The Internal Transcribed Spacers (ITS) region, elongation factor 1 alpha (EF1-α) and β-tubulin genes were amplified and sequenced using ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn, 1999), and Bt2a/Bt2b (Glass and Donaldson, 1995) primers respectively. The sequences of isolate MFT9 were deposited to GenBank (MW115977 (ITS), (MW118595 (EF1-α) and MW118596 (β-tubulin). All sequences showed more than 99.5% similarity with reported sequences of Lasiodiplodia theobromae isolate IBL340 with accessions numbers KT247466 (ITS), KT247472 (EF1-α) and KT247475 (β-tubulin). Phylogenetic reconstruction based on Maximum Likelihood, using Mega X (Kumar et al., 2018), grouped isolate MFT9 with isolates representing L. theobromae. Pathogenicity testing was performed on 18 fresh, healthy, medium-sized mango fruits for each treatment to fulfill Koch’s postulate. The fruits were disinfested with 1% NaClO and punctured with a sterilized needle to create approximately 2mm2 wounds for inoculation. Fruits were inoculated with 15µL of fresh inoculum (107 spores/mL) from isolate MFT9. Control fruits were inoculated with 15µL of SDW and both the inoculated and control fruits were incubated at 28°C for seven days of post inoculation. The rot lesions appeared at the point of inoculation and gradually spread on the fruit surface. The symptoms were similar to the symptoms observed on the original fruit samples (Fig. 2). This experiment was conducted three times under the same conditions, with control fruits remaining asymptomatic each time. The re-isolated fungus was identified as L. theobromae based on symptoms and morpho-molecular analysis, described above. L. theobromae is also reported as a causal agent responsible for a postharvest stem-end rot on Coconut in China (Zhang, et al., 2019). To our knowledge, this is the first report of L. theobromae causing postharvest stem-end rot of mango fruit in China. This finding suggests that L. theobromae is a potential problem for mango fruit production in China.

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