National Culture Collection Research Articles

Geographical Isolation and Root-Associated Fungi in the Marine Terrains: A Step Toward Establishing a Strategy for Acquiring Unique Microbial Resources

This study aimed to understand whether the geo-ecological segregation of native plant species affects the root-associated fungal community. Rhizoplane (RP) and rhizosphere (RS) fungal microbiota of Sedum takesimense native to three geographically segregated coastal regions (volcanic ocean islands) were analyzed using culture-independent methods: 568,507 quality sequences, 1399 operational taxonomic units, five phyla, and 181 genera were obtained. Across all regions, significant differences in the phyla distribution and ratio were confirmed. The Chao’s richness value was greater for RS than for RP, and this variance coincided with the number of genera. In contrast, the dominance of specific genera in the RS (Simpson value) was lower than the RP at all sites. The taxonomic identity of most fungal species (95%) closely interacting with the common host plant was different. Meanwhile, a considerable number of RP only residing fungal genera were thought to have close interdependency on their host halophyte. Among these, Metarhizium was the sole genus common to all sites. These suggest that the relationship between potential symbiotic fungi and their host halophyte species evolved with a regional dependency, in the same halophyte species, and of the same natural habitat (volcanic islands); further, the fungal community differenced in distinct geographical regions. Importantly, geographical segregation should be accounted for in national culture collections, based on taxonomical uniqueness.

Distribution of Pathogen Resources by the National Culture Collection for Pathogens in South Korea from 2015 to 2019.

Introduction: The National Culture Collection for Pathogens (NCCP) in the Korea Disease Control and Prevention Agency (KDCA) is in charge of the collection, quality control, and commercialization of pathogens and of information on pathogens that cause infectious diseases, and distributes them to be utilized for promoting public health and the health care industry. Since the NCCP Task Force was established in 2009 and the law for collection, management, and exploitation of pathogen resources became effective in 2017, NCCP has been responsible for the deposition, registration, and preservation of pathogen resources. Herein, we provide information on the recent status of pathogen resource distribution over the past 5 years and pathogen resources that are available for distribution presently. Results: Within the past 5 years, the numbers of pathogen resources distributed were 1659, 1823, 1457, 1156, and 1706 in the years 2015, 2016, 2017, 2018, and 2019, respectively. In addition, more pathogens were distributed in February, March, September, and October than in other months. When categorized by the type of institution, profit organizations (3069) requested more pathogen resources than national and public research institutes (2291). Furthermore, based on the purpose of distribution, 440 cases were for vaccine or therapeutic development, 3041 for diagnostic technology development, 2487 for education, and 1134 for quality control. Conclusion: Based on these findings, we publicized the information on the distribution of pathogen resources to encourage their widespread use in infectious disease research and public health.

Endophytic Fungi of Emersed Halophytes in River Deltas and Tidal Flats of the Korean Ramsar Wetlands

This study aimed to obtain information on the diversity and distribution of the endophytic fungi in Ramsar wetlands. Vast salt marshes in Suncheon Bay, Korea, are formed by two types of ecotones (tidal flats and deltas) that are supported by the emersed halophytes Phragmites australis and Suaeda japonica. Overall, 324 endophytes were isolated from P. australis (six sampling points in the delta and five in the tidal flats) and S. japonica (six in tidal flats). Margalef’s, Menhinick’s, Shannon’s, and Simpson’s diversity indices significantly varied among the ecotones. In particular, higher variance in diversity value and unevenness was observed in the delta marsh compared with the tidal flat marsh. Further, morphological diversity in the delta salt marsh was 1.8 times higher than that of the tidal flat. Comprising several dominant genera (Aspergillus, Cladosporium, and Penicillium), Epicoccum, Paraconiothyrium, Septoriella, and Talaromyces were widely distributed regardless of the aquatic conditions or halophyte species. This study highlights the distinct distribution and diversity of marine endophytes in various ecotones that can physically protect the coastal areas. This data is of much value to secure a national culture collection for future restoration of the coastal wetlands and their ecosystems.

The Israel National Culture Collection of Algae (INCCA) for biodiversity conservation

The aim of this paper is to introduce the Israel National Culture Collection of Algae (INCCA) http://kinneret. ocean. org. il/INCCA. aspx. INCCA was set up at the Kinneret Limnological Laboratory, Israel Oceanographic & Limnological Research, to document, isolate and preserve the microalgae biodiversity of Israel, for future scientific, industrial and educational purposes. At present the collection contains 103 strains, mainly from the Cyanobacteria, Chlorophyta and Charophyta divisions. Each isolated culture was first identified by microscopy and later submitted to DNA barcoding using rbcL and the rRNA SSU-ITS as markers. The unialgal (xenic) cultures are maintained on agar plates or in liquid media, at constant illumination and temperature. We present our isolation, purification and characterization procedures, together with a listing of our current holdings. The distinctiveness of INCCA among other culture collections is that it is the first collection of microalgae in Israel and its adjacent Middle Eastern countries. Furthermore, since most isolates are from Lake Kinneret they are accompanied by environmental data derived from >50 years of monitoring on Lake Kinneret. Examples of the research conducted on several of our strains supports the scientific importance of this collection. By transforming INCCA into a formal internationally accredited collection, that will expand to include species of microalgae from all types of environments in Israel, we would like to provide a home for preserving microalgal biodiversity in this Middle-East country.

First record of a marine microalgal species, <i>Jaagichlorella roystonensis</i> (Trebouxiophyceae) isolated from Jungmun Saekdal Beach, Jeju Island, Korea

A eukaryotic marine microalga was isolated from Jungmun Saekdal Beach, Jeju Island, Korea and an integrated approach, including molecular phylogeny and morphology, was used to determine its taxonomical status. Molecular phylogenetic evidence inferred from the small subunit (SSU) 18S rRNA sequence and internal transcribed spacer (ITS) secondary structure analysis clearly showed that the isolate belonged to the recently described species, Jaagichlorella roystonensis. Distinctive morphological keys of the species were also observed by light microscopy and scanning/ transmission electron microscopy (S/TEM). In this study, a Korean marine J. roystonensis species was described for the first time and was subsequently added to the national culture collections in Korea.

First record of a marine microalgal species, <i>Micractinium singularis</i> (Trebouxiophyceae) isolated from Janghang Harbor, Korea

A eukaryotic microalga was isolated from seawater in Janghang Harbor, Korea and its morphological, molecular, and physiological characteristics were investigated. Due to its simple morphology, no distinctive characters were found by morphological observation, such as light microscope or scanning/transmission electron microscopy (S/ TEM). However, molecular phylogenetic evidence inferred from the concatenated small subunit (SSU) 18S rRNA and internal transcribed spacer (ITS) sequence data indicated that the isolate belonged to the newly described Micractinium singularis. Furthermore, it was clustered with Antarctic Micractinium strains and it also showed a psychrotolerant property, surviving at temperatures as low as 5°C. However, its optimal growth temperatures range from 15°C to 25°C, indicating that this microalga is a mesophile. Additionally, gas chromatography-mass spectrometry (GC/MS) analysis showed that the isolate was rich in nutritionally important omega-3 polyunsaturated fatty acid, and high-performance liquid chromatography analysis (HPLC) revealed that the high-value antioxidant lutein was biosynthesized as an accessory pigment by this microalga, with glucose as the major monosaccharide. Therefore, in this study, a Korean marine M. singularis species was discovered, characterized, and described. It was subsequently added to the national culture collections.

Complete Genome Sequences of Micrococcus luteus Strains NCCP 15687 and NCCP 16831, Isolated in South Korea.

In this study, the complete genome sequences of Micrococcus luteus strains NCCP 15687 and NCCP 16831 were determined and deposited in the National Culture Collection for Pathogens (NCCP) of South Korea. Genomic DNA was isolated from blood samples from patients infected with M. luteus.

First record of a marine microalgal species, <i>Chlorella gloriosa</i> (Trebouxiophyceae) isolated from the Dokdo Islands, Korea

Chlorella gloriosa (Chlorellaceae, Trebouxiophyceae) was isolated from seawater off the coast of the Dokdo Islands in Korea. An axenic culture was established using the streak-plate method on f/2 agar media supplemented with antibiotics, allowing identification of the isolate by morphological, molecular, and physiological analyses. The morphological characteristics observed by light and electron microscopy revealed typical morphologies of C. gloriosa species. The molecular phylogenetic inference drawn from the small-subunit 18S rRNA sequence verified that the microalgal strain belongs to C. gloriosa. Additionally, gas chromatography-mass spectrometry analysis showed that the isolate was rich in nutritionally important omega-3 and -6 polyunsaturated fatty acids and high-performance liquid chromatography analysis revealed that the high-value antioxidants lutein and violaxanthin were biosynthesized as accessory pigments by this microalga, with arabinose, galactose, and glucose as the major monosaccharides. Therefore, in this study, a Korean marine C. gloriosa species was discovered, characterized, and described, and subsequently added to the national culture collection.

Borrelia maritima sp. nov., a novel species of the Borrelia burgdorferi sensu lato complex, occupying a basal position to North American species.

Borrelia species are vector-borne parasitic bacteria with unusual, highly fragmented genomes that include a linear chromosome and linear as well as circular plasmids that differ numerically between and within various species. Strain CA690T, which was cultivated from a questing Ixodes spinipalpis nymph in the San Francisco Bay area, CA, was determined to be genetically distinct from all other described species belonging to the Borrelia burgdorferi sensu lato complex. The genome, including plasmids, was assembled using a hybrid assembly of short Illumina reads and long reads obtained via Oxford Nanopore Technology. We found that strain CA690T has a main linear chromosome containing 902176 bp with a blast identity ≤91 % compared with other Borrelia species chromosomes and five linear and two circular plasmids. A phylogeny based on 37 single-copy genes of the main linear chromosome and rooted with the relapsing fever species Borrelia duttonii strain Ly revealed that strain CA690T had a sister-group relationship with, and occupied a basal position to, species occurring in North America. We propose to name this species Borrelia maritima sp. nov. The type strain, CA690T, has been deposited in two national culture collections, DSMZ (=107169) and ATCC (=TSD-160).

Efficient Utilization of Korean Medical Fungal Pathogen Resource Bank for Clinical Research

A "pathogen resource" contains information about pathogens (e.g., bacteria, fungi, viruses, and protozoa) and microbial derivatives (e.g., DNA, RNA, plasmid, clone, and cDNA). Pathogen resources are important for their potential use in healthcare research because they contain clinical and epidemiological information that is different from microbial resources. In October 2014, the "Nagoya Protocol" on access and benefit-sharing with the Convention on Biological Diversity was enacted to restrict the movement of transboundary pathogens and protect the natural pathogen resources of each country. On July 2017, the Korean Medical Fungal Pathogen Resource Bank (KMFRB) was established to secure, discover, and develop biological resources focused on medical fungi. KMFRB has since been operating under the National Culture Collection for Pathogens of the National Institute of Health based on the Act No. 13992. This report aims to provide general information regarding KMFRB and suggest efficient ways to utilize human fungal pathogen resources for clinical research.

Transport kinetics of chromium in perfused human placental lobule in late gestation: in vitro study

Objective: Reports relating to maternal-fetal transport kinetics of chromium, an essential trace element in the human pregnancies are scanty. Hence, we thought it interesting to investigate the transport kinetics of this trace element in the human placenta in late gestation in vitro.Methods: Human placentae were collected immediately after delivery from normal uncomplicated pregnancies. Chromium chloride solution (GFS Chem Inc, Ohio, USA) at 10 times the physiological concentrations and antipyrine (Sigma Chem Co., St. Louis, USA) as internal reference marker was injected as a single bolus (100 µl) into the maternal arterial circulation of perfused placental lobules and perfusate samples were collected from maternal and fetal circulations over a study period of 5 minutes. National culture and Tissue collection medium, diluted with Earle’s buffered salt solution was used as the perfusate. Serial perfusate samples were collected from fetal venous perfusate for a period of 30 minutes. Chromium concentration in perfusate samples was determined using atomic absorption spectrophotometry and the concentration of reference marker, antipyrine was measured by spectrophotometry. Transport kinetics and transport parameters of study and reference markers were assessed using well-established parameters.Results: Differential transport rates of chromium and antipyrine in 10 perfusions differed significantly for 10 and 50% efflux fractions (ANOVA test, p < .05) while those of 25, 75, and 90% efflux fractions were not significantly different between the study and reference substances. Chromium transport fraction (TF) averaged 54.9% of bolus dose in 10 perfusions while that of antipyrine averaged 89% of bolus dose, representing 61.80% of reference marker TF. The difference observed in TF values of chromium and antipyrine was statistically significant (Student’s t-test, p < .05). Pharmacokinetic parameters such as area under the curve, clearance, absorption rate, elimination rate of chromium compared to reference marker was significantly different (ANOVA test, p < .05) between the study and reference substances.Conclusions: Our studies report for the first time maternal-fetal transport kinetics of chromium in human placenta in vitro. Considering the restricted transfer of this essential trace element from maternal to fetal circulation despite its small molecular weight, we hypothesize an active transport of chromium across the human placental membrane. Further studies relating to placental transport kinetics of this trace element in various pregnancy-related disease states are in progress.

Assessment of the Growth and Fruiting of 19 Oyster Mushroom Strains for Indoor Cultivation on Lignocellulosic Wastes

AbstractTwelve Pleurotus ostreatus (Jacq. ex Fr.) P. Kumm and six Pleurotus pulmonarius (Fr.) Quel. strains were characterized from the National Culture Collection of Mushrooms, Institute of Botany ...

Prevalence of factor H Binding Protein sub-variants among Neisseria meningitidis in China

ObjectiveTo study the prevalence of the fHbp genes in Neisseria meningitidis (N. meningitidis) isolates for further evaluation and development of serogroup B meningococcal vaccines in China. MethodsA panel of 1012 N. meningitidis strains was selected from the national culture collection from 1956 to 2016, according to the years of isolation, locations, and strain sources. These were tested by FHbp variant typing. Multi-locus sequence typing (MLST) was performed on 822 of these samples, including 242 strains from clinical strains and 580 carrier-derived strains. Analysis based on sequence types, serogroups, and FHbp variations were used to summarize the prevalence and characteristics of N. meningitidis. ResultsThere were 8 serogroups of N. meningitidis as well as a collection of nongroupable strains in this study. 1008 of 1012 N. meningitidis strains tested were positive for the fHbp gene. Serogroup A N. meningitidis (MenA) strains belonging to ST-1 and ST-5 clonal complexes harbored genes only encoding variant 1 (v1) FHbp. All MenW strains encoded v2 FHbp. 61.9% of clinical MenB strains were positive for v2 FHbp vs. 32.1% that were positive for v1. Among fHbp-positive carrier-derived MenB strains, v2 FHbp accounted for 90.8%. 79.7% of clinical MenC strains were positive for v1 FHbp and 20.3% were positive for v2 FHbp. Among carrier-derived MenC strains, v2 FHbp predominated. The number of major serogroups of N. meningitidis analyzed by MLST was 822, and the encoded FHbp showed CC- or ST-specific characteristics. ConclusionfHbp genes were detected in almost all N. meningitidis strains in this study. Therefore, it is possible that a vaccine against MenB or meningococci irrespective of serogroups, which includes FHbp, could be developed. Meningococcal vaccine development for China is a complex issue and these findings warrant further attention with respect to vaccine development.

Candida glabrata species complex prevalence and antifungal susceptibility testing in a culture collection: First description of Candida nivariensis in Argentina.

The presence of the cryptic species belonging to the Candida glabrata complex has not been studied in Argentina. We analyzed a collection of 117 clinical isolates of C. glabrata complex belonging to a National Culture Collection of Instituto Nacional de Microbiología "Dr. Carlos G. Malbrán" from Argentina (40 isolates from blood samples, 18 from other normally sterile sites, 20 from vagina, 14 from urine, 7 from oral cavity, 3 from catheter, 1 from a stool sample and 14 isolates whose clinical origin was not recorded). The aims of this work were to determine the prevalence of the cryptic species Candida nivariensis and Candida bracarensis and to evaluate the susceptibility profile of isolates against nine antifungal drugs. Identification was carried out by using classical phenotypic tests, CHROMagar™ Candida, PCR and MALDI-TOF. The minimal inhibitory concentrations of amphotericin B, 5-fluorocytosine, fluconazole, itraconazole, voriconazole, ketoconazole, posaconazole, caspofungin and anidulafungin were determined according to the EDef 7.3 (EUCAST) reference document. Of the 117 isolates, 114 were identified as C. glabrata and three as C. nivariensis by using PCR and MALDI-TOF. There were no major differences between C. nivariensis and C. glabrata susceptibility profiles. No resistant strains were found to echinocandins. We have found that the percentage of C. nivariensis in our culture collection was 2.56. This is the first description of C. nivariensis in Argentina, and data obtained could contribute to the knowledge of the epidemiology of this cryptic species.

The U.S. Culture Collection Network Lays the Foundation for Progress in Preservation of Valuable Microbial Resources

The U.S. Culture Collection Network was formed in 2012 by a group of culture collection scientists and stakeholders in order to continue the progress established previously through efforts of an ad hoc group. The network is supported by a Research Coordination Network grant from the U.S. National Science Foundation (NSF) and has the goals of promoting interaction among collections, encouraging the adoption of best practices, and protecting endangered or orphaned collections. After prior meetings to discuss best practices, shared data, and synergy with genome programs, the network held a meeting at the U.S. Department of Agriculture (USDA)-Agricultural Research Service (ARS) National Center for Genetic Resources Preservation (NCGRP) in Fort Collins, Colorado in October 2015 specifically to discuss collections that are vulnerable because of changes in funding programs, or are at risk of loss because of retirement or lack of funding. The meeting allowed collection curators who had already backed up their resources at the USDA NCGRP to visit the site, and brought collection owners, managers, and stakeholders together. Eight formal collections have established off-site backups with the USDA-ARS, ensuring that key material will be preserved for future research. All of the collections with backup at the NCGRP are public distributing collections including U.S. NSF-supported genetic stock centers, USDA-ARS collections, and university-supported collections. Facing the retirement of several pioneering researchers, the community discussed the value of preserving personal research collections and agreed that a mechanism to preserve these valuable collections was essential to any future national culture collection system. Additional input from curators of plant and animal collections emphasized that collections of every kind face similar challenges in developing long-range plans for sustainability.

Halophilic bacteria mediated phytoremediation of salt-affected soils cultivated with rice

Salinity is a most important ecological constraint for cereal crop including rice nowadays. Phytoremediation has been used to remediate salt affected soil but the use of halophilic bacteria is limited. In this study, the salt resistant Thalassobacillus denorans (NCCP-58) and Oceanobacillus kapialis (NCCP (National Culture Collection of Pakistan) -76), halophilic bacteria isolated from salt mines near Karak, Pakistan were used to evaluate its effect on rice variety (Basmati-385)in soil contaminated with different concentrations (control, 50, 100 and 150mM) of NaCl. It has been observed that significant increase in germination percentage and germination rate occurred in seeds primed with bacterial strains as compared to non-primed seeds. Root and shoot lengths were more in plants raised from primed seeds than non-treated seeds. Plants raised from inoculated seeds showed a significant increase in fresh and dry weight of seedling after 15days. Significant increase in photosynthetic pigment; chlorophyll a (Chl a), chlorophyll b (Chl b) and carotenoid contents occurred in 28days old plants inoculated with bacterial strains under different saline conditions. Similarly significant increase occurred in total nitrogen and protein contents in inoculated plants. Ca+2 and K+ ion concentration significantly increased while Na+ ion concentration decreased in plants inoculated with bacterial strains as compared to non-inoculated plants under different saline conditions. Bacterial strains O. Kapialis (NCCP-76) was more responsive in term of physiological and biochemical parameters than T. denorans sp. (NCCP-58). The corresponding strains have a positive effect in alleviating the salt stress in plants growing in saline condition.

Antimicrobial Susceptibility and Characterization ofPropionibacterium acnesby Multilocus Sequence Typing and Repetitive-Sequence-Based PCR

Propionibacterium acnes, a gram-positive, anaerobic, and aerotolerant bacterium that is found frequently in the skin as part of the human microbiome causes inflammatory acne, shoulder infection, and the contamination of medical devices. The study goals were the antibiotic resistant and molecular epidemiological characterization of the P. acnes isolates in Korea. A total of 22 P. acnes isolates originated from diverse patients were obtained from three National Culture Collections for Pathogens in South Korea. The hemolytic properties and minimum inhibitory concentrations (MIC) of five antibiotics (tetracycline, doxycycline, clindamycin, erythromycin, and minocycline) were determined. Only one isolate showed high MIC values and resistance to all five antibiotics. Genotypic characterization was achieved by multilocus sequence typing (MLST) for eight loci (aroE, guaA, tly, camp2, atpD, gmk, lepA, and sodA) and repetitive-sequence-based PCR (rep-PCR) analysis using the DiversiLab kit. MLST revealed four phylogroups that were type IA 1 (27.3%), type IA 2 (18.2%), type IB (13.6%), and type II (40.9%). Rep-PCR results demonstrated three clusters that were cluster I (39.1%), cluster II (45.5%), and cluster III (13.6%). The isolates of cluster I were part of phylogroup type IA (both IA 1 and IA 2 ), and the isolates of cluster II belonged to phylogroup type II. All isolates of phylogroup type IB were hemolytic and belonged to cluster III. The results of rep-PCR clustering analysis showed a good correlation with those of MLST phylogroups, suggesting that rep-PCR could be an alternative method to track P. acnes subtype lineages.

Molecular identification of clinical Nocardia isolates from India.

The epidemiology of nocardiosis is evolving with increasing number of Nocardia spp. causing human infection. In recent years, molecular techniques have been used to identify Nocardia spp. There are limited data available on the spectrum of Nocardia spp. isolated from clinical samples in India. Here, a molecular study was carried on 30 clinical isolates maintained in our National Culture Collection to evaluate the techniques used for identifying the agents. The isolates were identified by sequencing two promising genes: the 16S rRNA gene and hsp65. Both hsp65 and the 16S rRNA gene could reliably identify 90 % of Nocardia isolates, i.e. N. farcinica, N. cyriacigeorgica, N. brasiliensis, N. otitidiscaviarum, N. amamiensis and N. pneumoniae. The mean percentage dissimilarity of sequence identification was higher using the hsp65 gene (4 %, range 0-7.9 %) compared with the 16S rRNA gene (2.3 %, range 0-8.9 %). Two isolates that showed ambiguous results in both the short segment of the 16S rRNA gene and hsp65 sequences could be resolved by sequencing a larger fragment (∼1000 bp) of the 16S rRNA gene. Both of these isolates were identified as N. beijingensis with similarities of 99.8 and 100 % compared with the standard strain. Genotyping of N. cyriacigeorgica strains was performed using hsp65 gene sequences and compared with previously described genotypes. Our N. cyriacigeorgica isolates belonged to genotype 1 (n = 4) and genotype 2 (n = 2). The present study highlights a wide spectrum of Nocardia spp. in India and emphasizes the need for molecular techniques for identification to the species level.

Maternal–fetal transport kinetics of manganese in perfused human placental lobule in vitro

Objective: There have been no detailed reports relating to maternal–fetal transport kinetics of manganese, an essential trace element in the human pregnancies, and hence we have attempted to study the transport kinetics of this trace element in the human placenta in vitro.Methods: Human placentae from normal uncomplicated pregnancies were collected postpartum. Manganese chloride solution (GFS Chem Inc., Columbus, OH), 10 times the physiological concentrations, along with antipyrine (Sigma Chem Co., St. Louis, MO) as reference marker were then injected as a single bolus (100 µl) into the maternal arterial circulation of perfused placental lobules and perfusate samples collected from maternal and fetal circulations over a period of five minutes. National Culture and Tissue Collection medium, diluted with Earle’s buffered salt solution was used as the perfusate and serial perfusate samples from fetal venous perfusate collected for a period of 30 min. Concentration of manganese in perfusate samples was assessed by atomic absorption spectrophotometry, while that of antipyrine was assessed by spectrophotometry. Transport kinetics of substances studied were computed using established permeation parameters.Results: Differential transport rates of manganese and antipyrine in 12 perfusions differed significantly for 25.75, 90% efflux fractions (ANOVA test, p < 0.05), while those of 10 and 50% efflux fractions were not significantly different between the study and reference substances. Transport fraction (TF) of manganese averaged 54.9% of bolus dose in 12 perfusions, whereas that of antipyrine averaged 89% of bolus dose, representing 61.80% of reference marker TF. The difference observed in TF values of manganese and antipyrine was statistically significant (Student’s t-test, p < 0.05). Pharmacokinetic parameters such as area under the curve, clearance, absorption rate, elimination rate of manganese compared to reference marker were significantly different (ANOVA test, p < 0.05) between the study and reference substances.Conclusions: Our studies show for the first time maternal–fetal transport kinetics of manganese in human placenta in vitro. Considering the restricted transfer of this essential trace element despite its small molecular weight, we hypothesize possibility of active transport of manganese across the human placental membrane. Further studies relating to manganese placental transport in “diabetic model” placental perfusions are in progress.

First Report of Soybean Sudden Death Syndrome Caused by Fusarium virguliforme in South Africa

Soybean (Glycine max (L.) Merr.) is an important crop in many countries and production is currently increasing (from 311,450 ha in 2010 to 516,500 ha in 2013) in South Africa. On 27 February 2013 in the Lydenburg/Badfontein area, Mpumalanga Province, on a no-till commercial farm planted to soybean cultivar PAN 737 (Roundup Ready, maturity group 7) under irrigation for a second consecutive season, leaf symptoms typical of soybean sudden death syndrome were observed and reported by a farmer (3). The symptoms developed at the R6 growth stage (near physiological maturity) of the soybean plants. Leaf symptoms were interveinal chlorotic blotches that became necrotic while the veins remained green. These symptoms appeared throughout the plant but were most severe on the top leaves. Some of the severely affected leaflets dropped off with the petioles remaining attached to the plant. The vascular tissue in the upper taproot and lower stem turned gray-brown, but the pith remained white. Roots of the affected plants had decayed lateral roots. Surface disinfested root pieces with rot symptoms and spores directly from blue sporodochia on the rotten root were plated on potato dextrose agar amended with novostreptomycin 0.04 g/L (PDA+). Slow growing Fusarium isolates with blue to purple masses of sporodochia were consistently obtained from diseased plants. Cultures were single-spored and plated on PDA+. Growth rate of cultures on PDA+ was on average 6 to 9 mm after 5 days at 20°C. The morphology of the isolates fit the description of Fusarium virguliforme in Aoki et al. (1). Sequence analyses of the nuclear ribosomal internal transcribed spacer (ITS) and partial translation elongation factor (EF-1a) gene of the recovered eight isolates revealed that these isolates matched 99.6% with F. virguliforme O'Donnell & T. Aoki (Accession Nos. KF648835 to KF648850), one of the soybean sudden death syndrome causing species found in North and South America (1). All isolates are identical in each loci except that three isolates had one nucleotide deletion and two insertions at the EF-1a loci. The isolates are deposited at the national culture collection in Pretoria (PPRI13434 to PPRI13441). A glasshouse bioassay was conducted to test the pathogenicity of eight single-spored isolates by inoculating pasteurized planting medium (1:1:1 ratio of sand, perlite, and soil) with a layer of infested sand-bran medium (2) to each pot (13 cm in diameter) and covered with 2 cm of planting medium (4) after planting 20 seeds of soybean cultivar PAN 737. There were three pots per isolate randomized in a complete block design trial. All the South African F. virguliforme isolates tested induced leaf and root rot symptoms of sudden death syndrome on the soybean seedlings under glasshouse conditions after 4 weeks of inoculation. The fungus was re-isolated on PDA+ from diseased roots of the soybean seedlings to fulfill Koch's postulates. This is the first record of F. virguliforme in South Africa, and as an important component of soilborne diseases of soybean it may pose a major threat to the South African soybean industry.