Sandalwood (Santalum album L; family Santalaceae) is a highly significant aromatic oil yielding tree. It is valued for two important traits, heartwood and essential oil obtained from the heartwood. This study was proposed to assess the morphological and genetic variability of sandalwood accessions. For this, genotypes were randomly selected (n = 177) from the 14 populations from three states in southern India. The total heartwood oil content and quality was estimated by UV method and GC-MS. Total 14 oil-specific genic SSR markers were procured to evaluate the genetic diversity among the sandalwood accessions. Total core size, heartwood content, and oil of S. album ranged from 4.4 to 19.1 cm; 0.0 to 17.3 cm; and 0.0 to 5.96% with covariance 27.61, 85.25, and 73.12% followed by mean 9.74, 3.77, and 2.71, respectively. Genetic diversity estimates were highly polymorphic in terms of Na 7.28, Ne 5.89, He 8.0 PIC 0.891, with little Ho, and F-0.922. AMOVA revealed that minimal genetic variation among populations and highest variation was found among individuals with Nm (58.4). The UPGMA reveals the cluster favored the grouping pattern by the PCA analysis. Structure and PCA analysis clustered the entire populations into two major groups with F ST 0.046 in which population of Kerala and Karnataka were pure and Telangana accessions were found admixtures. No significant correlation (r 2 = 0.23, P = 0.00) was observed between heartwood oil and genetic structures. A high degree of transferability of genic markers would facilitate the assessment of novel genotypes for future tree improvement and conservation of Sandalwood populations.
Extraction of DNA from fresh tissues is routine in studies of tropical forest species, but DNA extraction from wood is considered as difficult due to its highly degraded nature and adequate quality of genomic DNA extraction is essential for molecular studies. Very few studies have validated the potential for isolating DNA from dried wood (Heartwood and Sapwood). Wood genomic DNA extraction is difficult from mature timber (Teak (Tectona grandis f; verbanaceae), Black Rosewood (Dalbergia latifolia f; Fabaceae) Ben Teak (Lagerstroemia lanceolata f; Lytheraceae) tissues due to presence of high quantity of secondary metabolites polyphenols, tannins and terpenoids and protein inhibitors. Mostly in laboratories DNA extraction kits are available but by using kits, DNA yield is very low and it is quite expensive too. We have standardized and validated the DNA extraction through manual protocol which is applicable for Bark, Sapwood and Heartwood samples of tree species which contains huge amount of inflexible tissues and fibers. The quality of the DNA was tested by spectrophotometer, gel electrophoresis and PCR (ISSR and SSR) amplification. An avrage DNA yield for heartwood ranges from 0.186 -0.166 µg/µL and sapwood was ranges from 0.26 -0.244 µg/µL. Modification of CTAB method was by addition of polyvinylpyrrolidone (PVP) appx 0.25%, variation in Rnase concentration, proteinase treatment with different concentration and incubation time. In order to evaluate the standardized wood genomic DNA extraction protocol, we compared it with the mature leaf and core samples (heartwood and sapwood) of the same timber species. The outcome was also quantified and proved by means of polymerase chain reaction analysis by using ISSR and SSR microsatellite markers conducted with isolated pure DNAs. This modified protocol made increased yield and purity of wood total genomic DNA and facilitate the important application of forenHow to cite this paper: Fatima, T., Srivastava, A., Hanur, V.S. and Srinivasa Rao, M. (2018) An Effective Wood DNA Extraction
Japanese encephalitis is one of the serious vector-borne viral encephalitis diseases found worldwide and poses a major threat to public health. Most Japanese encephalitis virus (JEV) infections are subclinical; only 1: 250 to 1:1000 infected persons develop clinical presentations. Delay in proper diagnosis of JE affects the timeliness of treatment initiation and increases the mortality rate in patients. Therefore, there is an extreme need to develop potential biomarkers, which might improve the diagnosis and can become the basis for development of new therapeutics. The microRNAs (miRNAs/or miRs) are small noncoding RNAs of 17-24 nucleotides that are known to regulate about 60% of human genes. Although miRNAs have been found to regulate various aspects of innate and adaptive immune responses, less information on circulating miRNAs in JE is known. The study of JEV infected human serum miRNAs will provide novel information for the diagnosis of JE as well as for the improvement of disease outcome.Total RNA, including miRNA, was extracted from serum followed by the complementary DNA (cDNA) synthesis by using sequence-specific primers. cDNA was amplified using target-specific TaqMan MicroRNA Assay. Real-time polymerase chain reaction data was normalized using both exogenous (cel-miR-39) and endogenous (hsa-miR-93) controls.We have found significantly altered expression of miR-155 and miR-21 in serum of JEV infected patients as compared to healthy controls, revealing their role as a a noninvasive biomarker in JE. A significant correlation between miRNAs and JE was observed that offers the basis for miRNAs to serve as a new component to develop possible therapeutic strategies for JE in near future.
Sandalwood (Santalum album L.) is the second most expensive wood in the world. There are approximately 16 species of sandalwood (S.
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