BackgroundSpecies-specific microsatellite markers are desirable for genetic studies and to harness the potential of MAS-based breeding for genetic improvement. Limited availability of such markers for coffee, one of the most important beverage tree crops, warrants newer efforts to develop additional microsatellite markers that can be effectively deployed in genetic analysis and coffee improvement programs. The present study aimed to develop new coffee-specific SSR markers and validate their utility in analysis of genetic diversity, individualization, linkage mapping, and transferability for use in other related taxa.ResultsA small-insert partial genomic library of Coffea canephora, was probed for various SSR motifs following conventional approach of Southern hybridisation. Characterization of repeat positive clones revealed a very high abundance of DNRs (1/15 Kb) over TNRs (1/406 kb). The relative frequencies of different DNRs were found as AT >> AG > AC, whereas among TNRs, AGC was the most abundant repeat. The SSR positive sequences were used to design 58 primer pairs of which 44 pairs could be validated as single locus markers using a panel of arabica and robusta genotypes. The analysis revealed an average of 3.3 and 3.78 alleles and 0.49 and 0.62 PIC per marker for the tested arabicas and robustas, respectively. It also revealed a high cumulative PI over all the markers using both sib-based (10-6 and 10-12 for arabicas and robustas respectively) and unbiased corrected estimates (10-20 and 10-43 for arabicas and robustas respectively). The markers were tested for Hardy-Weinberg equilibrium, linkage dis-equilibrium, and were successfully used to ascertain generic diversity/affinities in the tested germplasm (cultivated as well as species). Nine markers could be mapped on robusta linkage map. Importantly, the markers showed ~92% transferability across related species/genera of coffee.ConclusionThe conventional approach of genomic library was successfully employed although with low efficiency to develop a set of 44 new genomic microsatellite markers of coffee. The characterization/validation of new markers demonstrated them to be highly informative, and useful for genetic studies namely, genetic diversity in coffee germplasm, individualization/bar-coding for germplasm protection, linkage mapping, taxonomic studies, and use as conserved orthologous sets across secondary genepool of coffee. Further, the relative frequency and distribution of different SSR motifs in coffee genome indicated coffee genome to be relatively poor in microsatellites compared to other plant species.

A series of β-carboline-linked 2,4-thiazolidinedione hybrids was synthesized and studied for their DNA affinities and cytotoxicities. The most potent compound was 19e with IC50 of 0.97 ± 0.13 μM.

A crucial event in calcium signaling is the transition of a calcium sensor from the apo (Ca free) to the holo (Ca-saturated) state. Caldendrin (CDD) is a neuronal Ca-binding protein with two functional (EF3 and EF4) and two atypical (EF1 and EF2), non-Ca-binding EF-hand motifs. During the transition from the apo to the holo state, guided by the stepwise filling of Ca, the protein passes through distinct states and acquires a stable conformational state when only EF3 is occupied by Ca. This state is characterized by a Ca-derived structural gain in EF3 with destabilization of the EF4 motif. At higher Ca levels, when Ca fills in EF4, the motif regains stability. EF3 controls initial Ca binding and dictates structural destabilization of EF4. It is likely that this unexpected intermotif communication will have an impact on Ca-dependent target interactions.

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