Shoot meristem culture eliminates bacterial and fungal infections from elite varieties of turmeric (Curcuma longa L.)

In Vitro Cellular & Developmental Biology - Plant - Turmeric (Curcuma longa L. (Zingiberaceae)) is a rich source of medicinally important chemical compounds obtained from both pseudostem...     

Whole Genomic Analysis of Human G12P[6] and G12P[8] Rotavirus Strains that Have Emerged in Myanmar

G12 rotaviruses are emerging rotavirus strains causing severe diarrhea in infants and young children worldwide. However, the whole genomes of only a few G12 strains have been fully sequenced and analyzed. In this study, we sequenced and characterized the complete genomes of six G12 strains (RVA/Human-tc/MMR/A14/2011/G12P[8], RVA/Human-tc/MMR/A23/2011/G12P[6], RVA/Human-tc/MMR/A25/2011/G12P[8], RVA/Human-tc/MMR/P02/2011/G12P[8], RVA/Human-tc/MMR/P39/2011/G12P[8], and RVA/Human-tc/MMR/P43/2011/G12P[8]) detected in six stool samples from children with acute gastroenteritis in Myanmar. On whole genomic analysis, all six Myanmarese G12 strains were found to have a Wa-like genetic backbone: G12-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1 for strains A14, A25, P02, P39, and P43, and G12-P[6]-I1-R1-C1-M1-A1-N1-T1-E1-H1 for strain A23. Phylogenetic analysis showed that most genes of the six strains examined in this study were genetically related to globally circulating human G1, G3, G9, and G12 strains. Of note is that the NSP4 gene of strain A23 exhibited the closest relationship with the cognate genes of human-like bovine strains as well as human strains, suggesting the occurrence of reassortment between human and bovine strains. Furthermore, strains A14, A25, P02, P39, and P43 were very closely related to one another in all the 11 gene segments, indicating derivation of the five strains from a common origin. On the other hand, strain A23 consistently formed distinct clusters as to all the 11 gene segments, indicating a distinct origin of strain A23 from that of strains A14, A25, P02, P39, and P43. To our knowledge, this is the first report on whole genome-based characterization of G12 strains that have emerged in Myanmar. Our observations will provide important insights into the evolutionary dynamics of spreading G12 rotaviruses in Asia.     

Combining flow cytometry and gfp reporter gene for quantitative evaluation of Pectpbacterium carotovorum ssp. carotovorum in Ornithogalum dubium plantlets

Aims: Ornithogalum dubium is a natural host of the soft rot pathogen Pectobacterium carotovorum ssp. carotovorum (Pcc). The present study was aimed to develop a quantification system for Pcc expressing a gfp reporter gene, using fluorescent activated cell sorter (FACS) in planta. Methods and Results: Several calibration steps were required to distinctly gate the GFP‐labelled bacteria at FL1 mode and count the bacteria. To validate the bacterial counts obtained by FACS analysis, an internal standard of polystyrene green fluorescent microsphere beads was employed, resulting in high correlation with serial dilutions and plate counting. This allowed quantification of the bacteria, with no further need to culture, dilute or plate the cells. Micropropagation tools were developed to produce uniform plantlets of O. dubium, which were either inoculated with increasing concentrations of Pcc or elicited for resistance towards Pcc using methyl jasmonate. The rapid counting procedure allowed recovering, gating and counting the bacterial population in planta, separately from the plant cells background and from the microsphere beads. Conclusions: The FACS based quantification approach of Pcc was found accurate, reproducible and time saving, thus useful for counting bacteria in planta. Significance and Impact of the Study: The combination of time‐ and cost‐saving approach for Pcc quantification with efficient screening tools during early stages of micropropagation may facilitate the preliminary process of selection for resistant cultivars.     

BrdU-Hoechst-Giemsa方法的进一步改进以及青鱼和草鱼复制带核型的研究

对BrdU-Hoechst-Giemsa方法进行了一定的改进和补充;并将它应用于青鱼和草鱼的核型研究中。实验进一步阐明,BrdU-Hoechst-Giemsa方法的关键性步骤之一是要先测算实验鱼的细胞周期;BrdU、AMD、Hoechst 33258,EB和AO虽都有抑制鱼类染色体浓缩、促进染色体分带的作用,但其中以DNA碱基特异性给合物BrdU、AMD和Hoechst 33258较佳,特别用AMD和Hoechst 33258同时处理活细胞的分带效果最好。     

Yet another improved silver staining method for the detection of proteins in polyacrylamide gels

Silver staining is very sensitive for detection of proteins in polyacrylamide gels and different procedures have been published. By combining and modifying some of the recipes, a very reproducible method, which is based upon staining with diamine complexes of silver, has been developed. The background staining is negligible and reduced silver does not precipitate on the gel surface. The technique works very well for sodium dodecyl sulfate-polyacrylamide gel electrophoresis in both homogeneous and in gradient gels as well as for two-dimensional (2-D) PAGE. It was possible to detect 1-10 ng of protein corresponding to approximately 50 pg/mm2, provided that a discontinuous buffer system was used, which gives sharp bands.     

Simple screening procedure for microorganisms to degrade amygdalin

Summary Amygdalin is a cyanogenic glycoside occurring among others in almonds and bitter apricot seeds. Utilization of seeds for human or animal nutrition requires adequate detoxification. The present method for screening microbial cultures for their ability to degrade amygdalin is a convenient micro-titre version based on the picrate reaction principle.     

Evaluation of the relationship among gene expressions and enzyme activities with antioxidant role and presenilin 1 expression in Alzheimer's disease

It is known that oxidative stress originating from reactive oxygen species plays a role in the pathogenesis of Alzheimer's disease. In this study, the role of antioxidant status associated with oxidative stress in Alzheimer's disease was investigated. Peripheral blood samples were obtained from 28 healthy individuals (as control) and 28 Alzheimer's patients who met the National Institute of Neurological and Communicative Diseases and Stroke/Alzheimer's Disease and Related Disorders Association criteria. Catalase, glutathione S-transferase and paraoxonase 1 enzyme activities in blood plasma and glutathione S-transferase enzyme activities in erythrocytes were determined by spectrophotometer. Catalase, glutathione S-transferase and presenilin 1 gene expressions in leukocytes were determined using qRT-PCR. Data were analysed with SPSS one-way anova , a LSD post hoc test at p < 0.05. The activity of each enzyme was significantly reduced in Alzheimer's patients compared to control. The catalase gene expression level did not change compared to the control. Glutathione S-transferase and presenilin 1 gene expression levels were increased compared to the control.     

Net Ecosystem Exchanges of Carbon, Water, and Energy in Young and Old-growth Douglas-Fir Forests

To be able to estimate the cumulative carbon budget at broader scales, it is essential to understand net ecosystem exchanges (NEE) of carbon and water in various ages and types of ecosystems. Using eddy-covariance (EC) in Douglas-fir dominated forests in the Wind River Valley, Washington, USA, we measured NEE of carbon, water, and energy from July through September in a 40-year-old stand (40YR) in 1998, a 20-year-old stand (20YR) in 1999, and a 450-year-old stand (450YR) during both years. All three stands were net carbon sinks during the dry, warm summers, with mean net daily accumulation of –0.30 g C m^–2 d^–1, –2.76 g C m^–2 d^–1, and –0.38 g C m^–2 d^–1, respectively, in the 20YR, 40YR, and 450YR (average of 1998, 1999) stands; but for individual years, the 450YR stand was a carbon source in 1998 (0.51 g C m^–2 d^–1) and a sink in 1999 (–1.26 g C m^–2 d^–1). The interannual differences for the summer months were apparent for cumulative carbon exchange at the 450YR stand, which had 46.9 g C m^–2 loss in 1998 and 115.9 g C m^–2 gain in 1999. As predicted, the 40YR stand assimilated the most carbon and lost the least amount of water to the atmosphere through evapotranspiration.