Transcriptome analysis of heat stress response genes in potato leaves.

Heat stress has a severe impact on potato growth and tuberization process, always resulting in the decrease of tuber yield and quality. Therefore, it is of great significance for potato breeding to illuminate the mechanism of heat stress on potato and explore heat resistant genes. In this study, two cDNA libraries from normal potato leaves (20 °C day/18 °C night) and potato leaves with 3 days of heat treatment (35 °C day/28 °C night) were constructed respectively. Totally, 1420 differentially expressed genes (DEGs) were identified. The expression patterns of 12 randomly selected genes detected using droplet digital PCR agreed with the sequencing data. Gene ontology analysis showed that these DEGs were clustered into 49 different GO types, reflecting the functional diversity of the heat stress response genes. The results of KEGG pathway enrichment showed the potential biological pathways in which the DEGs were involved, indicating that these pathways may be involved in heat tolerance regulation. Most potato heat transcription factors (StHsfs) and heat shock proteins (StHsps) were not expressed efficiently based on expression profile of these DEGs. StHsp26-CP and StHsp70 were markedly increased after 3 days of heat treatment. These data will be useful for further understanding the molecular mechanisms of potato plant tolerance to heat stress and provide a basis for breeding heat-tolerance varieties.

Characterization of natural melanin from Auricularia auricula and its hepatoprotective effect on acute alcohol liver injury in mice.

This study characterized the natural melanin from Auricularia auricula and investigated its hepatoprotective effect on mice with acute alcoholic liver injury. The characterization of the melanin was analyzed based on elemental analysis, gel permeation chromatography (GPC), UV-visible spectroscopy (UV-visible), infrared spectrum (IR) and nuclear magnetic resonance spectra (NMR). To determine the liver protective effect of Auricularia auricula melanin, mice were administered with the melanin once daily for 3 weeks before ethanol induced liver injury. Biochemical parameters of liver function, histopathological sections, mRNA and protein expression of antioxidant enzyme were compared between mice with or without the melanin administered. Results showed that A. auricula melanin was a eumelanin and the average molecular weight was 48.99 kDa. The melanin can protect the mice from ethanol-induced liver injury by extending the duration of the righting reflex, and shortening the duration of the recovery. The liver index, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (γ-GT) and liver malondialdehyde (MDA) levels in mice treated with the melanin were significantly decreased. At the same time, the levels of liver alcohol dehydrogenase (ADH), and antioxidase such as catalase (CAT), and superoxide dismutase (SOD) were increased. Its protective effect may be related to the activation of nuclear factor E2-related factor 2 (Nrf2) and its downstream antioxidant enzymes such as glutamate cysteine ligase catalytic (GCLC), glutamate cysteine ligase modifier (GCLM), and NADP(H) quinine oxidoreductase 1 (NQO-1). These results suggested that A. auricula melanin may be an effective strategy to alleviate alcohol-induced liver damage.

Analysis of the differential gene and protein expression profile of the rolled leaf mutant of transgenic rice (Oryza sativa L.).

The importance of leaf rolling in rice (Oryza sativa L.) has been widely recognized. Although several studies have investigated rice leaf rolling and identified some related genes, knowledge of the molecular mechanism underlying rice leaf rolling, especially outward leaf rolling, is limited. Therefore, in this study, differential proteomics and gene expression profiling were used to analyze rolled leaf mutant of transgenic rice in order to investigate differentially expressed genes and proteins related to rice leaf rolling. To this end, 28 differentially expressed proteins related to rolling leaf traits were isolated and identified. Digital expression profiling detected 10 genes related to rice leaf rolling. Some of the proteins and genes detected are involved in lipid metabolism, which is related to the development of bulliform cells, such as phosphoinositide phospholipase C, Mgll gene, and At4g26790 gene. The "omics"-level techniques were useful for simultaneously isolating several proteins and genes related to rice leaf rolling. In addition, the results of the analysis of differentially expressed proteins and genes were closely consistent with those from a corresponding functional analysis of cellular mechanisms; our study findings might form the basis for further research on the molecular mechanisms underlying rice leaf rolling.

Diversity and evolutionary relationship of nucleotide binding site-encoding disease-resistance gene analogues in sweet potato (Ipomoea batatas Lam.).

Most plant disease-resistance genes (R-genes) isolated so far encode proteins with a nucleotide binding site (NBS) domain and belong to a superfamily. NBS domains related to R-genes show a highly conserved backbone of an amino acid motif, which makes it possible to isolate resistance gene analogues (RGAs) by degenerate primers. Degenerate primers based on the conserved motif (P-loop and GLPL) of the NBS domain from R -genes were used to isolate RGAs from the genomic DNA of sweet potato cultivar Qingnong no.2. Five distinct clusters of RGAs (22 sequences) with the characteristic NBS representing a highly diverse sample were identified in sweet potato genomic DNA. Sequence identity among the 22 RGA nucleotide sequences ranged from 41.2% to 99.4%, while the deduced amino acid sequence identity from the 22 RGAs ranged from 20.6%to 100%. The analysis of sweet potato RGA sequences suggested mutation as the primary source of diversity. The phylogenetic analyses for RGA nucleotide sequences and deduced amino acids showed that RGAs from sweet potato were classified into two distinct groups--toll and interleukin receptor-1 (TIR)-NBS-LRR and non-TIR-NBS-LRR. The high degree of similarity between sweet potato RGAs and NBS sequences derived from R-genes cloned from tomato, tobacco, flax and potato suggest an ancestral relationship. Further studies showed that the ratio of non-synonymous to synonymous substitution within families was low. These data obtained from sweet potato suggest that the evolution of NBS-encoding sequences in sweet potato occur by the gradual accumulation of mutations leading to purifying selection and slow rates of divergence within distinct R-gene families.