Antisense therapeutics: is it as simple as complementary base recognition?

Antisense oligonucleotides provide a simple and efficient approach for developing target-selective drugs because they can modulate gene expression sequence-specifically. Antisense oligonucleotides have also become efficient molecular biological tools to investigate the function of any protein in the cell. As the application of antisense oligonucleotides has expanded, multiple mechanisms of oligonucleotides have been characterized that impede their routine use. Here, we discuss different mechanisms of action of oligonucleotides and the possible ways of minimizing non-antisense-related [corrected] effects to improve their specificity.     

Mechanisms, ecological consequences and agricultural implications of tri-trophic interactions

Recent research bridging mechanistic and ecological approaches demonstrates that plant attributes can affect herbivores, natural enemies of herbivores, and their interaction. Such effects may be genetically variable among plants and/or induced in individual plants by herbivore attack, and are mediated by primary plant attributes (i.e. nutritional quality and physical structure) and defense-related products (i.e. secondary chemicals and plant volatiles), and may be modified by human activity (e.g. by the introduction of Bacillus thuringiensis). The study of tri-trophic interactions is important in order to understand natural species interactions and to manipulate these interactions in pest control.     

E2F-1 is essential for normal epidermal wound repair.

E2F factors are involved in proliferation and apoptosis. To understand the role of E2F-1 in the epidermis, we screened wild type and E2F-1(-/-) keratinocyte mRNA for genes differentially expressed in the two cell populations. We demonstrate the reduced expression of integrins alpha(5), alpha(6), beta(1), and beta(4) in E2F-1(-/-) keratinocytes associated with reduced activation of Jun terminal kinase and Erk upon integrin stimulation. As a consequence of altered integrin expression and function, E2F-1(-/-) keratinocytes also show impaired migration, adhesion to extracellular matrix proteins, and a blunted chemotactic response to transforming growth factor-gamma1. E2F-1(-/-) keratinocytes, but not dermal fibroblasts, exhibit altered patterns of proliferation, including significant delays in transit through both G(1) and S phases of the cell cycle. Recognizing that proliferation and migration are key for proper wound healing in vivo, we postulated that E2F-1(-/-) mice may exhibit abnormal epidermal repair upon injury. Consistent with our hypothesis, E2F-1(-/-) mice exhibited impaired cutaneous wound healing. This defect is associated with substantially reduced local inflammatory responses and rates of re-epithelialization. Thus, we demonstrate that E2F-1 is indispensable for a hitherto unidentified cell type-specific and unique role in keratinocyte proliferation, adhesion, and migration as well as in proper wound repair and epidermal regeneration in vivo.     

Impact of ultraviolet-B radiation on photosynthetic capacity,antioxidative potential and metabolites in Solanum tuberosum L. under varying levels of soil NPK

The impact of supplemental ultraviolet-B (sUV-B; 280–315 nm; +7.2 kJ m^?2 d^?1) radiation was studied on various physiological parameters, antioxidative potential and metabolites of Solanum tuberosum L. cv. Kufri Badshah plants under varying levels of soil NPK. The N, P and K treatments were: the recommended dose of N, P and K; 1.5 times the recommended dose of N, P and K; 1.5 times the recommended dose of N and 1.5 times the recommended dose of K. The recommended NPK level provided maximum protection to photosynthetic assimilation under sUV-B radiation, while stomatal conductance was best at 1.5 times the recommended NPK. Carbon dioxide assimilation declined maximally at 1.5 times the recommended N/K under sUV-B radiation. Plants grown at the recommended NPK and 1.5 times the recommended NPK levels showed higher superoxide dismutase, peroxidase and ascorbate peroxidase activities under sUV-B radiation compared to 1.5 times the recommended N/K levels. sUV-B significantly increased total phenolics and flavonoids in plants at the recommended and 1.5 times the recommended NPK, while flavonoids declined at 1.5 times the recommended N. This study clearly showed that NPK amendment provided maximum protection to photosynthetic assimilation of potato plants under sUV-B radiation, activating the antioxidative defense system as well as flavonoids. NPK at 1.5 times the recommended dose, however, did not cause any additional benefit to photosynthetic carbon fixation; hence the recommended dose of NPK is found to be the best suited dose of fertilizer under ambient as well as sUV-B regime.     

Association mapping of agro-morphological characters among the global collection of finger millet genotypes using genomic SSR markers

Identification of alleles responsible for various agro-morphological characters is a major concern to further improve the finger millet germplasm. Forty-six genomic SSRs were used for genetic analysis and population structure analysis of a global collection of 190 finger millet genotypes and fifteen agro-morphological characters were evaluated. The overall results showed that Asian genotypes were smaller in height, smaller flag leaf length, less basal tiller number, early flowering and early maturity nature, small ear head length, and smaller in length of longest finger. The 46 SSRs yielded 90 scorable alleles and the polymorphism information content values varied from 0.292 to 0.703 at an average of 0.442. The gene diversity was in the range of 0.355 to 0.750 with an average value of 0.528. The 46 genomic SSR loci grouped the 190 finger millet genotypes into two major clusters based on their geographical origin by the both phylogenetic clustering and population structure analysis by STRUCTURE software. Association mapping of QTLs for 15 agro-morphological characters with 46 genomic SSRs resulted in identification of five markers were linked to QTLs of four traits at a significant threshold (P) level of ≤0.01 and ≤0.001. The QTL for basal tiller number was strongly associated with the locus UGEP81 at a P value of 0.001 by explaining the phenotypic variance (R ²) of 10.8 %. The QTL for days to 50 % flowering was linked by two SSR loci UGEP77 and UGEP90, explained 10 and 8.7 % of R ² respectively at a P value of 0.01. The SSR marker, FM9 found to have strong association to two agro-morphological traits, flag leaf width (P—0.001, R ²—14.1 %) and plant height (P—0.001, R ²—11.2 %). The markers linked to the QTLs for above agro-morphological characters found in the present study can be further used for cloning of the full length gene, fine mapping and their further use in the marker assisted breeding programmes for introgression of alleles into locally well adapted germplasm.     

A unique Ni2+ ‐dependent and methylglyoxal‐inducible rice glyoxalase I possesses a single active site and functions in abiotic stress response

The glyoxalase system constitutes the major pathway for the detoxification of metabolically produced cytotoxin methylglyoxal (MG) into a non‐toxic metabolite d ‐lactate. Glyoxalase I (GLY I) is an evolutionarily conserved metalloenzyme requiring divalent metal ions for its activity: Zn²⁺ in the case of eukaryotes or Ni²⁺ for enzymes of prokaryotic origin. Plant GLY I proteins are part of a multimember family; however, not much is known about their physiological function, structure and metal dependency. In this study, we report a unique GLY I (OsGLYI‐11.2) from Oryza sativa (rice) that requires Ni²⁺ for its activity. Its biochemical, structural and functional characterization revealed it to be a monomeric enzyme, possessing a single Ni²⁺ coordination site despite containing two GLY I domains. The requirement of Ni²⁺ as a cofactor by an enzyme involved in cellular detoxification suggests an essential role for this otherwise toxic heavy metal in the stress response. Intriguingly, the expression of OsGLYI‐11.2 was found to be highly substrate inducible, suggesting an important mode of regulation for its cellular levels. Heterologous expression of OsGLYI‐11.2 in Escherichia coli and model plant Nicotiana tabacum (tobacco) resulted in improved adaptation to various abiotic stresses caused by increased scavenging of MG, lower Na⁺/K⁺ ratio and maintenance of reduced glutathione levels. Together, our results suggest interesting links between MG cellular levels, its detoxification by GLY I, and Ni²⁺ – the heavy metal cofactor of OsGLYI‐11.2, in relation to stress response and adaptation in plants.