Bioactive Peptides from Algae: Traditional and Novel Generation Strategies,Structure-Function Relationships,and Bioinformatics as Predictive Tools for Bioactivity

Over the last decade, algae have been explored as alternative and sustainable protein sources for a balanced diet and more recently, as a potential source of algal-derived bioactive peptides with potential health benefits. This review will focus on the emerging processes for the generation and isolation of bioactive peptides or cryptides from algae, including: (1) pre-treatments of algae for the extraction of protein by physical and biochemical methods; and (2) methods for the generation of bioactive including enzymatic hydrolysis and other emerging methods. To date, the main biological properties of the peptides identified from algae, including anti-hypertensive, antioxidant and anti-proliferative/cytotoxic effects (for this review, anti-proliferative/cytotoxic will be referred to by the term anti-cancer), assayed in vitro and/or in vivo, will also be summarized emphasizing the structure–function relationship and mechanism of action of these peptides. Moreover, the use of in silico methods, such as quantitative structural activity relationships (QSAR) and molecular docking for the identification of specific peptides of bioactive interest from hydrolysates will be described in detail together with the main challenges and opportunities to exploit algae as a source of bioactive peptides.     

Suitability of mapped sequence tagged microsatellite site markers for establishing distinctness, uniformity and stability in aromatic rice

At present, testing for distinctness, uniformity and stability (DUS) of crop varieties relies on a set of morphological characters. These characters suffer fromthe limitations of number, interaction with the environment in which the variety grows and subjectivity in decision-making. The potential of DNA-based markers such as sequence tagged microsatellite site (STMS), for establishing DUS merits investigation. In the present study, a set of 55 mapped STMS markers, selected from 12 linkage groups of rice genome, was used to examine distinctness of 23 aromatic rice genotypes including the commercially important Basmati varieties. Forty-one of these markers (74.5%) showed polymorphism between the varieties. The number of alleles per locus ranged from 2–4 with an average of 2.3. The polymorphism information content (PIC) of the markers varied from 0.083 to 0.665 with an average of 0.338. All the varieties could be differentiated from each other at a low probability (0.07×10^-13) of identical match by chance. The marker-based clustering of the varieties corresponded with the known phenotypic classification, thereby providing confidence in the distinctness established by the mapped STMS markers. The utility of these markers to study uniformity and stability was analysed using a commercially important crossbred Basmati rice variety Pusa Basmati 1(IET-10364) that contributes about 40–50% of Basmati rice export from India. Genotyping of twenty individual plants, grown from the nucleus, breeder, foundation, certified and farmer's saved seed samples using all the 55 markers revealed no variation among the plants. These observations suggested that the set of mapped markers employed in this study could be further used for establishing distinctness of aromatic rice varieties and for studying DUS of the important commercial variety Pusa Basmati 1. This revised version was published online in July 2006 with corrections to the Cover Date.     

One-step RT-PCR for the Detection of Infectious Bursal Disease Virus in Clinical Samples

A single-tube, non-interrupted, one-step RT-PCR has been standardized to amplify the hypervariable region of the VP2 gene sequence of infectious bursal disease virus (IBDV). The technique standardized on purified viral RNA was successfully applied to the detection of the virus directly in clinical samples. The amplified products were confirmed to be IBDV specific by their size in ethidium bromide-stained agarose gel, nested PCR and restriction enzyme digestion. Digestion of the amplicons with StyI restriction enzyme also differentiated classical virus from six very virulent field isolates. The sensitivity of the one-step RT-PCR was found to be 0.2 pg of viral RNA.     

Evaluation of hyperimmune sera against goat pox viral antigens

Tropical Animal Health and Production -     

Molecular genetic mapping of quantitative trait loci associated with loaf volume in hexaploid wheat (Triticum aestivum)

Major efforts in wheat research are being made to improve the yield and quality of wheat. Loaf volume (Lv) is the main quality parameter deciding the bread making potential of wheat. To genetically dissect quantitative trait loci (QTLs) for Lv, a Recombinant Inbred Line (RIL) population (F₈) was developed from a cross between two Indian wheat varieties “HI 977” and “HD 2329”. A total of 914 SSR and 100 ISSR primers were used for molecular analysis and the genetic map comprising 19 chromosomes was constructed with 202 SSR markers and 2 HMW glutenin subunit loci: Glu-B1 and Glu-D1. The phenotypic data were collected from six environments including three different agro-climatic zones for 2 consecutive years. Dissection of Lv through AMMI model revealed significant G×E variance for the trait. QTL analysis was performed using composite interval mapping. A total of 30 QTLs for Lv were detected and significant QTLs were identified on 6B and 6D chromosomes; 1B, 1D, 2A, 3A, 5B and 5D also contributed genetically to Lv. Association between 6B and 6D QTLs and variable expression of gliadins on group 6 chromosomes were discussed. QTLs detected in this study were compared with other QTL analysis in wheat.     

Microbial populations,enzyme activities and nitrogen-phosphorus-potassium enrichment in earthworm casts and in the surrounding soil of a pineapple plantation

Summary Total populations of bacteria and fungi, dehydrogenase activity (as a measure of total potential microbial activity), and urease and phosphatase activities were determined in earthworm casts and surrounding laterite soils planted to pineapple. The casts contained higher microbial populations and enzyme activities than the soil. Except for fungal populations, statistically significant (P = 0.05) increases were found in all other parameters. Microbial populations and enzyme activities showed similar temporal trends with higher values in spring and summer and lower values in winter. The earthworm casts contained higher amounts of N, P, K and organic C than the soil (P = 0.05). Selective feeding by earthworms on organically rich substrates, which break down during passage through the gut, is likely to be responsible for the higher microbial populations and greater enzyme activity in the casts.     

Protection of mice against <Emphasis Type="Italic">Brucella abortus 544</Emphasis> challenge by vaccination with recombinant OMP28 adjuvanted with CpG oligonucleotides

Brucella abortus, a gram negative, facultative intracellular pathogen causes brucellosis in many animal species and humans. Although live, attenuated vaccines are available against this infection, they suffer from certain limitations. Therefore, the development of an effective subunit vaccine against brucellosis is an area of intense research. The outer membrane proteins (OMPs) of Brucella species have been extensively studied for its immunogenicity and protective ability. We have investigated the potential of CpG ODN to enhance the immunogenicity and protective efficacy of recombinant 28 kDa outer membrane protein (rOMP28) of Brucella melitensis. The study demonstrated vigorous immunoglobulin G (IgG) response of OMP28. The administration of rOMP28 with CpG caused increased cell mediated immune response in terms of induced IgG2a, T-cell proliferation and up-regulation of type I cytokine expression. In contrast, the free antigen suppressed the interferon gamma (type I cytokine) production on in-vitro stimulation of spleenocytes. The result indicates the role of OMP28 in the down regulation of IFN-γ production. Moreover, the B. abortus S-19 vaccinated mice showed highest production of IL-4 and IFN-γ. The protective ability of the antigen was evaluated by systemic bacterial clearance after challenging the mouse with B. abortus 544 pathogen. The level of protection was significant in rOMP28+CpG treated mice but was lower than the required level. The results of the present study indicate that rOMP28 could be an immunogen capable of inducing both humoral and cellular immune response. The humoral response was biased towards Th1 type when it was co-administered with CpG.     

Path-Coefficient Analysis of Leaf-Anatomical Characters Affecting Frost Injury in Potato

Correlation and path-coefficient analysis for leaf-anatomical characters affecting frost injury was carried out in twenty eight varieties and accessions belonging to several wild and cultivated tuber-bearing species of Solanum in order to identify selection criteria in breeding for frost resistance in potato Palisade thickness and palisade proportion showed a significant negative correlation with frost injury Path-coefficient analysis revealed that palisade thickness had the highest direct negative effect on frost injury whereas leaf thickness, not conforming to its behaviour as found in simple correlation analysis, emerged ax the next highest factor to have direct positive effect on frost injury. Direct and indirect effects of these factors on frost injury vis a vis their interdependence were found to be in support ot the ‘palisade thickness method’ for screening frost resistant genotypes in potato breeding programmes.