Determination of cytogenetic and epigenetic effects of manganese and copper on Zea mays L.

Heavy metal accumulation and its possible effects are prominent problem for not only human health but also for the environment and plant systems due to that heavy metals are non-biodegradable. In this research, it was aimed to examine the impacts of heavy metals on toxicity and genotoxicity in maize. Seeds of corn were subjected to various concentrations of MnSO4 and CuSO4 for determining their effects on DNA methylation, DNA damage levels, protein and phytohormone alterations. The results revealed that an increase in copper and manganese concentrations causes decrease in soluble protein levels, genomic template stability (GTS) and mitotic index but causes an increase in RAPD profile alterations and DNA hypermethylation. Additionally, HPLC analyses show that CuSO4 and MnSO4 contamination reduces growth-promoting hormones, like gibberellic acid (GA), zeatin (ZA) and indole acetic acid (IAA), and increases the abscisic acid (ABA). This study obviously indicated that CuSO4 and MnSO4 have epigenetic and genotoxic effects. A decrease in the phytohormone level (ZA, GA, and IAA) and an increase in the ABA level under CuSO4 and MnSO4 are thought to be a part of the defense system of maize to struggle with stress.

Assessment of Genetic Diversity in Promising Sugarcane Mutants Developed Through Chemical Mutagenesis

Sugarcane is an important cash crop gaining importance as ideal raw material for bioenergy production. Conventional sugarcane improvement is carried out through intensive crossing and selection breeding within the Saccharum complex. Being highly polyploidy crop the mutation breeding is also integrated for specifically improving the desired genetic trait in sugarcane varieties developed through conventional breeding. In the present work RAPD markers based assessment of genetic diversity and kinship relationships of six promising mutants developed using CoC 671 has been reported. The banding pattern of the PCR amplified products showed polymorphism with the RAPD profile in all six mutants. Mutants CoC 671, TC 2813, TC 2819, TC 2826, TC 2875 formed a separate cluster, and TC 906, TC 922, and TC 906 B formed a separate cluster distinct from the earlier one with the parent (CoC 671) and the standard Check (Co 86023). The genetic distance between the groups was found to be only marginally higher than the respective group distances and the overall mean genetic distance. The similarity indices indicated that there was 15 % genetic dissimilarity between CoC 671 and Co 86032. This may be because CoC 671 was one of the parents for Co 86032. In the present investigation, the genetic similarity value ranged from 0.97 to 0.78 among the mutants. A total of 34 band positions were scored, out of which 15 RAPD bands were polymorphic. From the genetic similarity coefficient based on RAPD band data sharing, it was found that the majority of the clones were almost more than 78% similar to the mother plant. Thus the level of similarity between varieties seemed to be not very high but the mutants could be distinguished from each other easily. RAPD analysis indicated EMS-induced point mutations resulting in specific rectifications without much change in the genetic backbone of genotype CoC 671. Since this analysis was carried out on mutants after multiplying in field for three successive generations, we presume that these mutants/genotypes are stable and express the stable genetic variations.

Characterization of Clinical MRSA Isolates from Northern Spain and Assessment of Their Susceptibility to Phage-Derived Antimicrobials

Methicillin-resistant Staphylococcus aureus (MRSA) is a prevalent nosocomial pathogen, causing a wide range of diseases. The increased frequency of MRSA isolates in hospitals and the emergence of vancomycin resistance have sparked the search for new control strategies. This study aimed to characterize sixty-seven MRSA isolates collected from both infected patients and asymptomatic carriers in a Spanish hospital. RAPD-PCR allowed the identification of six genetic patterns. We also investigated the presence of genes involved in producing adhesins, toxins and the capsule; the biofilm; and antimicrobial resistance. A notable percentage of the isolates carried virulence genes and showed medium-high ability to form biofilms. Next, we assessed the strains’ susceptibility to two phages (phiIPLA-C1C and phiIPLA-RODI) and one endolysin (LysRODI). All strains were resistant to phiIPLA-C1C, and most (70.2%) were susceptible to phiIPLA-RODI. Regarding LysRODI, all strains displayed susceptibility, although to varying degrees. There was a correlation between endolysin susceptibility and the random amplification of polymorphic DNA (RAPD) profile or the presence of some virulence genes (fnbA, eta, etb, PVL and czr), but that was not observed with biofilm-forming ability, strain origin or phage sensitivity. Taken together, these findings can help to explain the factors influencing endolysin effectiveness, which will contribute to the development of efficient therapies targeting MRSA infections.

Mutation determination of rice by using RAPD primers

PCR is a powerful tool for the amplification of genetic sequences but sometimes, even though using an established PCR protocol that had been optimized and successful for the amplification of a particular DNA segment, use of that same protocol on a different region can result in a less than desirable outcome. Therefore, an experiment was conducted at Molecular biology laboratory of Malaysian Nuclear Agency during December 2016 to January 2017 used seeds of 6 indica rice cultivars. To conduce RAPD experiments for the rice species it was established the following reaction conditions for the final volume of 20 μl where 0.1 unit of Taq DNA polymerase, 0.4 μl of each dNTP, 2.5 mM MgCl2, 1 μl primer and 2.0 μl of DNA template. From this experiment, it is clear that after mutation the parent MR219 performed some genetic modification and produce genetically different variety namely NMR151, NMR152, ML3, ML10 and ML30. These mutant varieties have two different groups based on their mutation source and it is clear enough from their RAPD profile. Int. J. Agril. Res. Innov. & Tech. 9 (1): 1-7, June, 2019

In vitro Multiplication and Genetic Fidelity Studies in Cleopatra Mandarin (Citrus reshni Tanaka)

The present investigation was undertaken for in vitro multiplication of Cleopatra mandarin using nodal segment explants. Nodal segments taken from 9 years old plant in the month of March - April were cultured on Murashige and Skoog (1962) medium supplemented with BAP 0.5 mg/l + Kin 0.5 mg/l for establishment. Various concentrations of auxins viz. IAA and NAA were used singly or in combination with cytokinins (Kinetin and BAP) for shoot multiplication. Maximum number of shootlets per sprouted bud (8.2 ± 0.22) and maximum length of shootlets (3.0 ± 0.12cm) were recorded when auxins were used in combination with cytokinins at a concentration of NAA 0.3 mg/l + BAP 1.0 mg/l. Full strength MS media fortified with 1mg/l IBA was used for rooting of multiplied shoots and was kept in potting media containing sand, soil and vermi compost in 1:1:1 ratio for hardening.In vitro raised plants were examined for genetic stability by using RAPD primers. Out of fifty primers screened, eleven primers produced amplification while thirty-nine primers did not show any amplification. All RAPD profile for in vitro raised plants were monomorphic and similar to their mother plant, which showed that all the plants raised through micropropagation were true to type.

Random amplified polymorphic DNA-based molecular heterogeneity analysis of Salmonella enterica isolates from foods of animal origin

Aim: This study aims to study the significance of random amplified polymorphic DNA (RAPD) typing in heterogeneity analysis of Salmonella serovars, isolated from foods of animal origin. Materials and Methods: Salmonella serovars isolated and identified from different foods of animal origin such as meat, milk, and egg by standard bacteriological methods. DNA isolated from all 10 isolates which are confirmed by biochemical and serotyping methods and then RAPD was performed using the primers OPB 10, primer 1290, NSC I, NSC II, and primer 3. Then, RAPD data were analyzed using the BioNumerics software, Belgium, Germany. Results: RAPD polymerase chain reaction (PCR) using five primers, namely OPB 10, primer 1290, NSC I, NSC II, and primer 3, classified the 10 isolates into 9, 10, 10, 7, and 10 RAPD-PCR types with discriminating powers of 0.1987, 0.423, 0.50889, 0.1842, and 0.2582, respectively. The phylogram constructed with NSC I profile classified isolates based on geographical origin. Primer 1290, NSC II, and primer 3 produced some uniform bands in all isolates indicating their binding ability in conserved genomic region. This study revealed that RAPD profile can be best used for finding out the heterogeneity at molecular level of Salmonella isolates in combination with other molecular and phenotypic typing techniques. Thus, our results support earlier observation of its significance by different workers on different Salmonella serotypes. Conclusion: Repeatability of RAPD-PCR is insufficient to distinguish genetic differences among Salmonella serovars.

Aeromonas hydrophila associated with mass mortality of adult Goldfish, Carassius auratus (Linnaeus 1758) in ornamental farms in India

Aeromonas hydrophila was identified the causative agent of a disease outbreak in goldfish, Carassius auratus from four ornamental fish farms in Kerala, India. Seven bacterial isolates viz., Plesiomonas shigelloides (NPPS-1), Aeromonas hydrophila (NPAH-1, 2, 3 and 4), Citrobacter freundii (NPCF-1) and Acinetobacter spp. (NPA-1) were isolated from the affected fish. Further, all four A. hydrophila (NPAH-1, 2, 3 and 4) isolates were identified by amplification of gyrB and rpoD genes. The RAPD profile using 3 primers confirmed that all four A. hydrophila were genetically similar. No cytopathic effect was observed on goldfish fin (GFF) cell line after inoculation of the tissue homogenate from the affected fish and affected tissues were found negative for koi herpesvirus (KHV), cyprinid herpesvirus-2 (CyHV-2) and spring viraemia of carp virus (SVCV). Experimental challenge resulted in mortality of fish injected with A. hydrophila only. A. hydrophila was observed to be cytotoxic on GFF cell line and exhibited haemolytic activity on 5% sheep blood agar. A. hydrophila possessed multiple virulence genes viz., enterotoxins, haemolytic toxins and outer membrane protein as determined by PCR. A. hydrophila was sensitive to Cefixime, Chloramphenicol, Nitrofurantoin, Kanamycin, Ciprofloxacin, Furazolidone and Cefixime/Clavulanic acid. After treatment with the suggested antibiotics, the fish were recovered from the disease.

Common Ancestry & Genetic Diversity of Few Indigenous Chilli Land Races of North East India

Since chilies contain high quantities of bioactive potential compounds, they are expected to possess large number of pharmacological values like antioxi-dants, antifungal, antibacterial, anti-inflammatory, antiulcer, diuretics activi-ties and is being employed for the treatment of different ailments in the indigenous system of medicine. The current study aimed at comparative evaluation of antioxidant properties, phytochemicals and RAPD assay from extracts of 6 different types of chilli found in the states of Assam and Arunachal Pradesh of North East India. The plant extracts were screened for phyto-chemicals, antioxidant analysis and RAPD profile. Phytochemical analysis shows abundant presence of alkaloids, phenols, flavonoids, saponins etc. The antioxidants activities of all the aqueous extracts were evaluated using the DPPH method. The antioxidant activity (% oxidation inhibition) ranged from as high as 86.56% in Bhut Jolokia (Capsicum assamicum) extracts to as low as 18.77% in Badami Jolokia (Capsicum annum) extract. Chilli samples studied possess potential health benefits by inhibiting many oxidation reactions caused by free radicals. Genetic diversity in 6 chilli genotypes was analyzed by RAPD markers of the fourteen random primers among which 11 were polymorphic. The phylogenetic tree showed common ancestry and the divergence among the chilli species. Based on the current study findings, it may be concluded the RAPD technology can be a useful tool in explaining the genetic diversity among chilli genotypes.