In recent years, there has been a great deal of attention toward free radicals, reactive oxygen species (ROS) generated by exposure of crop plant cells to physical radiations. Henceforth, the current study was planned to compare oxidative stress and mutagenic potential of different irradiation doses of fast neutron (FN) and UV-B on meiotic-pollen mother cells (PMCs), pollen grains (PGs) and seeds yielded from irradiated faba beans seedlings. On the cytogenetic level, each irradiation type had special interference with DNA of PMC and exhibited wide range of mutagenic action on the frequency and type of chromosomal anomalies, fertility of PGs and seed yield productivity based on the irradiation exposure dose and radiation sensitivity of faba bean plants compared with un-irradiated ones. On the molecular level, SDS-PAGE and RPAD-PCR analyses of seeds yielded from irradiated seedlings exhibited distinctive polymorphisms based on size, intensity, appearance, and disappearance of polypeptides bands compared with un-irradiated ones. The total values of protein and DNA polymorphisms reached 88% and 90.80% respectively. The neutron fluency (2.3 × 106 n/cm2) and UV-B dose for 1 hr were recorded as bio-positive effects. The present study proved that genetic variations revealed by cytogenetic test could be supported by gene expression (alterations in RAPD and protein profiles).
ABSTRACT. In this study, we analyzed the correlation between genetic variation based on random amplified polymorphic DNA (RAPD), acid phosphatase, and glutamate-oxaloacetate transaminase isozymes, and amino acid composition with the antioxidant potential status of 7 wild Trigonella foenum-graecum L. accessions collected from diverse ecogeographical regions. RAPD revealed that 90 DNA products had highly polymorphism value (94.12%) based on band numbers, with sizes ranging from 50-2100 base pairs, and band intensity. Of 49 DNA polymorphic bands, 31 unique and 3 monomorphic bands were scored. Acid phosphatase and glutamate-oxaloacetate transaminase showed total polymorphism values of 90.00 and 93.75%, respectively, based on zymogram number, relative front (R f ), and optical intensity. Because isozymes are composed of amino acids, they were analyzed using highperformance liquid chromatography, which revealed the presences of 16 amino acids of variable content ranging from 13.21-15.35%, 9 of which are essential amino acids in humans. RAPD and isozymes showed similarly high estimates of genetic variability. Genetic relationships RAPD, isozymes, and antioxidant activity of fenugreek accessions revealed by unweighted pair group method with arithmetic mean clustering analysis based on data obtained from all primers of RAPD and each isozyme were very similar. The antioxidant potential based on free radical scavenging, 2, 2-diphenyl-1-picrylhydrazyl, b-carotenelinoleate, total phenolic, and flavonoid contents values were variable among accessions. We found that fenugreek is a valuable genetic resource with high antioxidant activity. Their genotypes, based on data and clustering of RAPD, isozymes, and variable amino acid contents, combined with their antioxidant potential statues are important in fenugreek breeding and improvement programs.
ABSTRACT. Protein and DNA polymorphismswere surveyed among seven accessions of wild fenugreek (Trigonellafoenum-graecum L.) to estimate their genetic diversity and relationships. Samples were obtained from diverse ecogeographical areas in Saudi Arabia and Yemen. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of seed storage protein showed genetic variations among fenugreek germplasms, both quantitatively and qualitatively, generating a total of 168 polypeptide bands with different molecular weights ranging from 4.5 to 300 kDa. Twenty-six of these bands were polymorphic, with a considerable polymorphism value (80.00%). Furthermore, restriction fragment length polymorphism (RFLP) analysis was also employed, which was based on the ability of four restriction enzymes (EagI, EcoRI, FspI, and HindIII) to cleave genomic DNA of the plant materials at specific target nucleotide sequences into different SDS-PAGE and RFLP analyses of fenugreek accessions numbers of DNA fragments. RFLP analysis revealed 166 fragments with known sequences and variable lengths ranging from 80 to 4000 bp with a highly degree of polymorphism (88.71%). Data derived from SDS-PAGE or RFLP analyses were used to produce dendrograms, which clustered the studied fenugreek accessions into different groups based on the unweighted pair group method with arithmetic mean (UPGMA). The resulting relationships indicated that these two marker techniques were nearly equivalent, but not identical, with respect to phylogenetic information. In conclusion, SDS-PAGE analysis of seed proteins should be augmented with RFLP analysis of DNA for reliable estimates of genetic diversity among fenugreek germplasms.
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