Identification and characterization of important sterile and maintainer lines from various genotypes for advanced breeding programmes of onion (Allium cepa)

Onion is one of the major vegetable crops in terms of production as well as consumption. In the current research, available onion genetic stock was evaluated to identify male-sterile lines and produce high-yielding F₁ hybrids for future breeding programmes. A mitochondrial DNA-based marker was mapped and correlated with phenotypic traits to isolate male-sterile plants. Based on the floral and pollen structure, nine putative male-sterile lines were identified. On the other hand, for nuclear marker identification at Ms locus, two sets of primers were used, one for Ms dominant allele and another for sterile and maintainer plants. Results revealed that 70% of open pollinated varieties (OPVs) possess plants with sterile cytoplasm coupled with genetic sterility at Ms locus, called sterile “A” line. Approximately 20% of plants in some genotypes were identified with normal (N) cytoplasm having recessive fertility gene at Ms locus, called maintainer “B” line. Based on the present findings, “A”, “B” and “R” (restorer line), future F₁ hybrid seed production systems in onion is discussed.     

Purification and characterization of glycerolipid acyl-hydrolase from the red alga Gracilaria vermiculophylla

ABSTRACT:   A glycerolipid acyl-hydrolase was purified 19-fold with a yield of 11% from the prostaglandin-producing red alga Gracilaria vermiculophylla by ammonium sulfate precipitation, anion-exchange chromatoraphy and gel filtration chromatography. Sodium dodecylsulfate– polyacrylamide gel electrophoresis of the final preparation showed a single band corresponding to a molecular mass of 20 kDa, but Superdex 200 fast protein liquid chromatography exhibited a molecular mass of 40 kDa. Accordingly, it was suggested that the purified enzyme was a homodimer of a 20 kDa subunit. The optimal temperature and pH were 37°C and 7–8, respectively. The purified enzyme catalyzed hydrolysis of the acyl groups of both glycoglycerolipids and phospholipids, especially monogalactosyldiacylglycerol and phosphatidylcholine. These results suggest that the enzyme hydrolyze the membrane lipids of the alga to release various saturated and unsaturated fatty acids, including arachidonic acid as substrate for prostaglandin synthesis.     

Identification of Wap65, a human homologue of hemopexin as a copper-inducible gene in swordtail fish, <Emphasis Type="Italic">Xiphophorus helleri</Emphasis>

Copper is one of the major heavy metal pollutants found in the aquatic environment. Therefore, it is important for determining the genes that play a key role in copper metabolism in aquatic organisms. This study, thus, aimed to identify a new copper-inducible gene in swordtail fish, Xiphophorus helleri. Using ACP-based RT-PCR coupled with RLM-RACE, we cloned Wap65, a mammalian homologue of hemopexin gene. The gene exhibits high identity at amino acid levels with the Wap65 gene of other fish species (42–68%) and mammalian hemopexin gene (35–37%). In addition, ten cysteine and two histidine residues are conserved in the swordtail fish Wap65 gene. These cysteine residues are vital for structural integrity, and histidine residues provide high binding affinity towards heme. As revealed by RT-PCR, the gene was upregulated in swordtail fish that were exposed to copper in a dose- and time-dependent manner. Therefore, the identification of Wap65, a mammalian homologue of hemopexin, as a new copper-inducible gene will provide greater insight into the role of this gene in copper metabolism.     

Antioxidant Activity of Hydrolysates and Peptide Fractions of Nemipterus japonicus and Exocoetus volitans Muscle

The focus of the study was to investigate the antioxidant activity of hydrolyzed muscle protein of Nemipterus japonicus and Exocoetus volitans. The trypsin protein hydrolysates of both fish showed maximum free radical scavenging potential and lipid peroxidation inhibition. Furthermore, it was purified by chromatographic methods followed by the lipid peroxidation inhibition; free radical scavenging assay was performed before and after purification. The purified peptide fractions of N. japonicus and E. volitans exhibited higher activity against polyunsaturated fatty acids (PUFA) peroxidation which was similar to natural antioxidants like α-tocopherol. Free radical scavenging potencies were measured by electron spin resonance technique (ESR). The purified peptide of E. volitans quenched free radicals (DPPH, hydroxyl, and superoxide) slightly more than N. japonicus. The amino acid composition of both fish protein hydrolysates showed variations in their ratio. The purified peptides were tested for cell cytotoxicity for Vero (kidney epithelial cells of the African Green Monkey) and Hep G₂ (human hepatocellular liver carcinoma) cell lines. It was found that peptides did not show any cytotoxic effect for Vero cell lines and exerted a significant antiproliferative effect on Hep G₂ cell lines.