CRISPR/Cas9 genome editing systems have been established in a broad range of eukaryotic species. Herein, we report the first method for genetic engineering in pyogo (shiitake) mushrooms (Lentinula edodes) using CRISPR/Cas9. For in vivo expression of guide RNAs (gRNAs) targeting the mating-type gene HD1 (LeA1), we identified an endogenous LeU6 promoter in the L. edodes genome. We constructed a plasmid containing the LeU6 and glyceraldehyde-3-phosphate dehydrogenase (LeGPD) promoters to express the Cas9 protein. Among the eight gRNAs we tested, three successfully disrupted the LeA1 locus. Although the CRISPRCas9–induced alleles did not affect mating with compatible monokaryotic strains, disruption of the transcription levels of the downstream genes of LeHD1 and LeHD2 was detected.Based on this result, we present the first report of a simple and powerful genetic manipulation tool using the CRISPR/Cas9 toolbox for the scientifically and industrially important edible mushroom, L. edodes.

A pandemic influenza A (H1N1) virus strain was isolated from a pig farm in Korea in December 2009. The strain was propagated in and isolated from both the Madin-Darby canine kidney cell line and embryonated eggs. The partial and complete sequences of the strain were identical to those of A/California/04/2009, with >99% sequence similarity in the HA, NA, M, NS, NP, PA, PB1, and PB2 genes. The isolated strain was inactivated and used to prepare a swine influenza vaccine. This trial vaccine, containing the new isolate that has high sequence similarity with the pandemic influenza A (H1N1) virus, resulted in seroconversion in Guinea pigs and piglets. This strain could therefore be a potential vaccine candidate for swine influenza control in commercial farms.
Agriculture ; Animals ; Cell Line ; Eggs ; Guinea Pigs ; Influenza Vaccines ; Influenza, Human* ; Kidney ; Korea* ; Orthomyxoviridae* ; Ovum ; Pandemics* ; Seroconversion ; Swine

Sixteen genomic DNA simple sequence repeat (SSR) markers of Lentinula edodes were developed from 205 SSR motifs present in 46.1-Mb long L. edodes genome sequences. The number of alleles ranged from 3–14 and the major allele frequency was distributed from 0.17–0.96. The values of observed and expected heterozygosity ranged from 0.00–0.76 and 0.07–0.90, respectively. The polymorphic information content value ranged from 0.07–0.89. A dendrogram, based on 16 SSR markers clustered by the paired hierarchical clustering' method, showed that 33 shiitake cultivars could be divided into three major groups and successfully identified. These SSR markers will contribute to the efficient breeding of this species by providing diversity in shiitake varieties. Furthermore, the genomic information covered by the markers can provide a valuable resource for genetic linkage map construction, molecular mapping, and marker-assisted selection in the shiitake mushroom.
Alleles ; Breeding ; DNA* ; Gene Frequency ; Genetic Linkage ; Genetic Variation ; Genome ; Lentinula* ; Methods ; Microsatellite Repeats ; Shiitake Mushrooms*

Plummer-Vinson syndrome is characterized by dysphagia, iron deficiency anemia, and upper esophageal web. The associated symptoms can be resolved by administering iron supplements as well as by endoscopic intervention. Relapse in patients with Plummer-Vinson syndrome is very rare. We describe a case of a 42-year-old woman with Plummer-Vinson syndrome whose symptoms were successfully treated with endoscopic dilatation and iron supplementation at first admission; however, 1 year later, she revisited our hospital because of dysphagia. On second admission, investigations revealed esophageal web relapse in Plummer-Vinson syndrome. She was again successfully treated with endoscopic dilatation and iron supplementation. After first admission, her anemia was not normalized due to poor compliance and loss of follow-up. We experienced a case of esophageal web relapse due to uncorrected iron deficiency anemia in a patient with Plummer-Vinson syndrome. This experience indicates that continuous iron supplementation and long-term follow-up is important in patients with Plummer-Vinson syndrome.
Adult ; Anemia ; Anemia, Iron-Deficiency ; Compliance ; Deglutition Disorders ; Dilatation ; Female ; Follow-Up Studies ; Humans ; Iron ; Plummer-Vinson Syndrome* ; Recurrence

Plummer-Vinson syndrome is characterized by dysphagia, iron deficiency anemia, and upper esophageal web. The associated symptoms can be resolved by administering iron supplements as well as by endoscopic intervention. Relapse in patients with Plummer-Vinson syndrome is very rare. We describe a case of a 42-year-old woman with Plummer-Vinson syndrome whose symptoms were successfully treated with endoscopic dilatation and iron supplementation at first admission; however, 1 year later, she revisited our hospital because of dysphagia. On second admission, investigations revealed esophageal web relapse in Plummer-Vinson syndrome. She was again successfully treated with endoscopic dilatation and iron supplementation. After first admission, her anemia was not normalized due to poor compliance and loss of follow-up. We experienced a case of esophageal web relapse due to uncorrected iron deficiency anemia in a patient with Plummer-Vinson syndrome. This experience indicates that continuous iron supplementation and long-term follow-up is important in patients with Plummer-Vinson syndrome.
Adult ; Anemia ; Anemia, Iron-Deficiency ; Compliance ; Deglutition Disorders ; Dilatation ; Female ; Follow-Up Studies ; Humans ; Iron ; Plummer-Vinson Syndrome* ; Recurrence