The Osmotin-Like Protein Gene PdOLP1 Is Involved in Secondary Cell Wall Biosynthesis during Wood Formation in Poplar.

Osmotin-like proteins (OLPs) mediate defenses against abiotic and biotic stresses and fungal pathogens in plants. However, no OLPs have been functionally elucidated in poplar. Here, we report an osmotin-like protein designated PdOLP1 from Populus deltoides (Marsh.). Expression analysis showed that PdOLP1 transcripts were mainly present in immature xylem and immature phloem during vascular tissue development in P. deltoides. We conducted phenotypic, anatomical, and molecular analyses of PdOLP1-overexpressing lines and the PdOLP1-downregulated hybrid poplar 84K (Populus alba × Populus glandulosa) (Hybrid poplar 84K PagOLP1, PagOLP2, PagOLP3 and PagOLP4 are highly homologous to PdOLP1, and are downregulated in PdOLP1-downregulated hybrid poplar 84K). The overexpression of PdOLP1 led to a reduction in the radial width and cell layer number in the xylem and phloem zones, in expression of genes involved in lignin biosynthesis, and in the fibers and vessels of xylem cell walls in the overexpressing lines. Additionally, the xylem vessels and fibers of PdOLP1-downregulated poplar exhibited increased secondary cell wall thickness. Elevated expression of secondary wall biosynthetic genes was accompanied by increases in lignin content, dry weight biomass, and carbon storage in PdOLP1-downregulated lines. A PdOLP1 coexpression network was constructed and showed that PdOLP1 was coexpressed with a large number of genes involved in secondary cell wall biosynthesis and wood development in poplar. Moreover, based on transcriptional activation assays, PtobZIP5 and PtobHLH7 activated the PdOLP1 promoter, whereas PtoBLH8 and PtoWRKY40 repressed it. A yeast one-hybrid (Y1H) assay confirmed interaction of PtoBLH8, PtoMYB3, and PtoWRKY40 with the PdOLP1 promoter in vivo. Together, our results suggest that PdOLP1 is a negative regulator of secondary wall biosynthesis and may be valuable for manipulating secondary cell wall deposition to improve carbon fixation efficiency in tree species.

Molecular epidemiology and virulence characteristics of Staphylococcus aureus nasal colonization in medical laboratory staff: comparison between microbiological and non-microbiological laboratories.

BACKGROUND: Medical laboratory staff are a high-risk population for colonization of Staphylococcus aureus (S. aureus) due to direct and dense contact with the pathogens; however, there is limited information about this colonization. This study sought to determine the prevalence and molecular characteristics of nasal colonization by S. aureus in medical laboratory staff in Guangzhou, southern China, and to compare the differences between microbiological laboratory (MLS) and non-microbiological laboratory (NMLS) staff. METHODS: S. aureus colonization was assessed by nasal swab cultures from 434 subjects, including 130 MLSs and 304 NMLSs from 33 hospitals in Guangzhou. All S. aureus isolates underwent the antimicrobial susceptibility test, virulence gene detection and molecular typing. RESULTS: The overall prevalence of S. aureus carriage was 20.1% (87/434), which was higher in MLSs than in NMLSs (26.2% vs. 17.4%, P < 0.05), while the prevalence of Methicillin-resistant S. aureus (MRSA) was similar. Living with hospital staff was associated with S. aureus carriage. The majority of the isolates harboured various virulence genes, and those in MLSs appeared less resistant to antibiotics and more virulent than their counterparts. A total of 37 different spa types were detected; among these, t338, t437, t189 and t701 were the most frequently encountered types. T338 was the main spa type contributing to nasal colonization Methicillin-sensitive S. aureus (MSSA) (13.0%), and t437-SCCmec IV was predominant in MRSA isolates (40%). CONCLUSIONS: These findings provide insight into the risk factors, molecular epidemiology and virulence gene profiles of S. aureus nasal carriage among the medical laboratory staff in Guangzhou.

Molecular epidemiology and characteristic of virulence gene of community-acquired and hospital-acquired methicillin-resistant Staphylococcus aureus isolates in Sun Yat-sen Memorial hospital, Guangzhou, Southern China.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of both hospital and community infections globally. It's important to illuminate the differences between community-acquired MRSA (CA-MRSA) and hospital-acquired MRSA (HA-MRSA), but there have been confusions on the definition, especially for the MRSA isolates identified within 48 h of admission. This study aimed to determine the molecular characteristics and virulence genes profile of CA and HA-MRSA isolates identified less than 48 h after hospital admission in our region. METHODS: A total 62 MRSA isolates identified within 48 h after admission and the clinical data were collected. Antimicrobial susceptibility test (AST) of collected isolates were performed according to the guidelines of Clinical and Laboratory Standards Institute (CLSI) 2015, and staphylococcal cassette chromosome mec (SCCmec) typing, multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) and virulence gene profiling were performed to explore the molecular diversity. RESULTS: SCCmec III and sequence type (ST) 239 were the most prevalent SCCmec type and ST in both CA and HA-MRSA groups. HA-MRSA group had higher prevalence of SCCmec III (87.2 %) and ST239 (79.5 %) compared with CA-MRSA (60.9 and 43.4 %, both P < 0.001), while the frequency of SCCmec IV (26.0 %) and ST59 (21.7 %) were higher in CA-MRSA than its counterpart (P < 0.001 and P = 0.003). MRSA-ST239-III was the predominant type in this study (61.3 %, 38/62), especially in HA-MRSA group (76.9 %, 30/39). However, CA-MRSA strains exhibited more diversity in genotypes in this study. Meanwhile, CA-MRSA tended to have lower resistant percentage to non-ß-lactams antibiotics but more virulence genes carriage, especially the staphylococcal enterotoxins (SE) genes. Notably, seb gene was only detected in CA-MRSA isolates (52.2 %), likely a significant marker for CA-MRSA isolates. Panton-Valentine leukocidin gene (PVL) was highly detected in both groups, while appeared no significantly different between CA-MRSA (47.8 %) and HA-MRSA (43.6 %). CONCLUSIONS: Our findings support a difference in the molecular epidemiology and virulence genes profile of CA-MRSA and HA-MRSA. Furthermore, this study indicates a possible transmission from HA-MRSA to CA-MRSA, which may cause the overlap of the definition.

Serotype distribution and antibiotic resistance of Streptococcus pneumoniae isolates collected at a Chinese hospital from 2011 to 2013.

BACKGROUND: Streptococcus pneumoniae infections are a major cause of global morbidity and mortality, and the emergence of antibiotic-resistant Streptococcus pneumoniae strains has been increasingly reported. This study provides up-to-date information on bacterial serotype distribution and drug resistance from S. pneumoniae clinical isolates that could guide prevention and treatment strategies for pneumococcal disease in China. METHODS: A total of 94 S. pneumoniae isolates were collected from outpatients and inpatients at one Chinese hospital from 2011-2013. Drug susceptibility and resistance was determined by minimum inhibitory concentrations (MICs). Capsular serotypes were identified by the quellung reaction test and multiplex polymerase chain reaction. RESULTS: Fifteen serotypes were identified among the 94 S. pneumoniae clinical isolates that were collected. Prevalent serotypes were 19F (42.6 %), 19A (8.5 %), 3 (8.5 %), and 6B (7.4 %). Potential immunization coverage rates for the 7-, 10- and 13-valent pneumococcal polysaccharide conjugate vaccines were 59.6, 62.6, and 79.6 %, respectively. Resistance rates to tetracycline, erythromycin, and trimethoprim/sulfamethoxazole were 91.2, 80.2 and 63.8 %, respectively. Resistance rates to penicillin, amoxicillin, ceftriaxone, and cefotaxime were 47.3, 34.1, 19.8, and 18.7 %, respectively. In almost all cases, antimicrobial resistance of the S. pneumoniae isolates in patients five years or younger was higher than isolates collected from patients aged 51 years or older. CONCLUSION: Prevalent serotypes among the 94 S. pneumoniae clinical isolates were 19F, 19A, 3, and 6B. The 13-valent pneumococcal polysaccharide conjugate vaccine covered the majority of the serotypes identified in this sample. Drug resistance varied among different serotypes and age groups. Clinical precautions should be taken to avoid the development of multidrug resistance in this potential human pathogen.

Differences in Staphylococcus aureus nasal carriage and molecular characteristics among community residents and healthcare workers at Sun Yat-Sen University, Guangzhou, Southern China.

BACKGROUND: The pathogenic potential and commensal nature of Staphylococcus aureus allows for easy transmission both within and outside of the hospital environment, and nasal carriage may be responsible for some serious infections. This study aimed to determine the molecular and epidemiological characteristics of nasal colonization by S. aureus in community residents (CR) and healthcare workers (HW) at Sun Yat-Sen University, Guangzhou, China. METHODS: A total of 589 volunteers, both CR (n = 297) and HW (n = 292), were recruited. Each subject completed a questionnaire, and specimens were obtained from the anterior nares for S. aureus screening. Genotypic analysis included pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), staphylococcal cassette chromosome mec (SCCmec) typing, and virulence gene detection. RESULTS: A total of 138 S. aureus isolates were recovered from separate subjects (23.4%, 138/589), with four isolates showing methicillin resistance (0.7%, 4/589). The prevalence of S. aureus carriage was 25.3% (75/297) in CR and 21.6% (63/292) in HW. Methicillin-resistant S. aureus (MRSA) were isolated from one CR (0.3%, 1/297) and three HW (1.0%, 3/292). The most common risk factors for S. aureus carriage in CR were being male, age ≤ 30 years, and nasal cavity cleaning habits. Having a household member in the healthcare profession was associated with increased risk among HW. Sequence type (ST)188 and ST59 were the most prevalent among the 20 observed STs, accounting for 14.6% and 12.2% of all isolates, respectively. The four MRSA isolates presented four different STs, with one isolate carrying a type IVa SCCmec element and the other three isolates containing type III SCCmec. PFGE analysis grouped the 129 isolates into 23 pulsotypes, with profiles A, N, E, L, and O the most prevalent. The Panton-Valentine leucocidin gene (pvl) was identified in two of the 138 isolates, while 57.5% of isolates carried both the Staphylococcus aureus enterotoxin A (sea) and enterotoxin B (seb) genes. CONCLUSIONS: These data indicate a low prevalence of nasal MRSA carriage but evidence of molecular heterogeneity among S. aureus isolates from CR and HW at Sun Yat-Sen University, Guangzhou. Differences in epidemiological and molecular characteristics of S. aureus between CR and HW populations may be useful for the understanding and prevention of S. aureus infection.