Potential utility of 14 candidate housekeeping genes as normalization reference for RT-qPCR analysis with developmental samples (fertilized eggs to late veliger larvae) in Pacific abalone Haliotis discus hannai was evaluated using four different statistical algorithms (geNorm, NormFinder, BestKeeper and comparative ΔCT method). Different algorithms identified different genes as the best candidates, and geometric mean-based final ranking from the most to the least stable expression was as follow: RPL5, RPL4, RPS18, RPL8, RPL7, UBE2, RPL7A, GAPDH, RPL36, PPIB, EF1A, ACTB and B-TU. The findings were further validated via relative quantification of metallothionein (MT) transcripts using the stable and unstable reference genes, and expression levels of MT were greatly influenced according to the choice of reference genes. In overall, our data suggest that RPL5 and RPS18, either singly or in combination, are appropriate for normalizing gene expression in developmental samples of this abalone species, whereas ACTB, B-TU and EF1A are less stable and not recommended. In addition, our findings propose that standard deviations in geometric ranking as well as geometric mean itself should also be taken into account for the final selection of reference gene(s). This study could be a useful basis to facilitate the generation of accurate and reliable RT-qPCR data with developmental samples in this abalone species.
The black-veined white, Aporia crataegi (Lepidoptera: Papilionoidea), is nearly extinct in South Korea, although substantial numbers of dried specimens are available. One of the common practices for such species is to launch re-introduction program after proper amount of genetic information are analyzed from donor and donee populations. In this study, we sequenced complete mitochondrial genome (mitogenome) of A. crataegi to design species-specific primers for subsequent population works and to further understand the mitogenome evolution in lepodiopteran Papilionoidea. The 15,140-bp long A. crataegi mitogenome that has typical sets of 37 genes is smallest among true butterfly species with overall slightly smaller size in genes and regions throughout the genome. Arrangement of the genome is identical to those of other lepidopteran mitogenomes, in which tRNA cluster located between the A+T-rich region and ND2 gene is translocated into tRNAMet, tRNAIle, and tRNAGln from ancestral arrangement, tRNAIle, and tRNAGln, tRNAMet. The A/T content of the genome at 81.3% is the highest in Pieridae, but lower than that of lycaenid species (81.7% ~ 82.7%) The high A/T content in the genome is also reflected in codon usage, accounting for 41.69% of A/T-composed codons (TTA, ATT, TTT, and ATA). Unlikely the diversified or modified usage of anticodon for tRNASer(AGN) the species of Pieridae including A. crataegi all unanimously have GCT that has been hypothesized as ancestral for Lepidoptera. A total of 111 bp of non-coding sequences are dispersed in 13 regions, ranging in size from 1–49 bp. Among them relatively longer ones (≥ 16 bp) all have relatively higher sequence identity to other regions of the genome, suggesting partial duplication of the sequences during A. crataegi evolution. As has been reported in some species of Lepidoptera, the A. crataegi A+T-region also has typically found conserved sequences (e.g., poly-T stretch, ATAGA motif, ATTTA element, microsatellite-like A/T sequence, and poly-A stretch) and one tRNA-like sequence, and this feature was commonly found in true butterfly species.
Antioxidant activity is important for reducing oxidative stress that causes various metabolic disorders. Metabolic disorders are highly related to loss of productivity in livestock. Therefore, development of effective antioxidant compounds originating from plants is important for organic agriculture. Phenolic compounds in edible plants are regarded as major components relevant to antioxidant activity. The present study investigated the changes in antioxidant activity and phenolic compound profiles of Aronia (Aronia meloncarpa) by fermentation using different strains of Leuconostoc mesenteroides. A total of 5 strains of L. mesenteroides were used as starter cultures and their β-glucosidase activities were measured. A total of 6 experiment runs were prepared, one for control (uninoculated) and the others (inoculated) for treatments. For biological activity, antioxidant and antibacterial activities were measured. For phenolic compound profiling, TLC and HPLC analysis were performed. The strains of KACC12313 and KACC12315 showed greater enzyme activity than others. Treatment with KCCM35046 showed strong and broad antibacterial activity against to Listeria monocytogenes. Treatments with KCCM35046 and KACC12315 showed the highest total polyphenol content. The highest antioxidant activity was found in KACC12315 treatment. No remarkable alteration was found in thin layer chromatography (TLC) analysis. In phenolic compound profiling analysis, KCCM35046 showed notable alteration in compound area ratio compared to others and also showed the highest caffeic acid content. In chlorogenic acid, treatments with KCCM35046 and KACC12315 showed great content than others. Treatment with KACC12315 showed the greatest content of trans-ferulic acid. As a result of relative performance indexing analysis, L. mesenteroides KCCM35046 and KACC12315 were selected as the best strain for the fermentation of Aronia.
Geneally, rice seeds regardless indica or japonica are showing low germination ratio or completely lost germination ability together with lost of good eating quality under high temperature and humidity conditions. Thus, this study was designed to evaluate a longevity for conservation of good eating quality during long term storage in rice. For the longevity evaluation, germination ability was studied after 5 days of high temperature and humidity stress (50℃/RH 95%). Dharial, originated from Bangladesh and showing weedy type with red pericarp, was selected as a good donor for longevity genes. A mutant was developed from Dharial through EMS mutagenesis and two populations of Dharial/4*Ilmibyeo and Dharial/4*Gopumbyeo were also developed for genetic study. In the 2-DE analysis followed by MALDI-TOF MS with wild and mutant lines, several candidate genes were identified. In the longevity test of two populations, a few lines showing good germination ability after high temperature and humidity stress were selected and subjected to confirm the relationships between longevity and conservation of good eating quality under long term storage.