Sweet orange (Citrus sinensis), a key fruit species, is considered as a primary ingredient in herbal medical formulations against ailments such as food borne diseases. Sour orange (C. aurantium) is also very famous as a medicinal plant. There are six commonly grown sweet orange cultivars in Sri Lanka (Arogya, Bibila sweet, MKD, Sisila, BAN and MT) but the antibacterial activity present in their fruit juice is not well documented. Therefore, the present study was conducted to characterize the antibacterial activity of the fruit juice of these sweet oranges in comparison to sour orange and also to establish DNA barcodes for the tested cultivars for precise identification. Fruit juice was collected from sweet orange cultivars and sour orange and antibacterial activity was measured against three model pathogenic bacterial species, Escherichia coli, Staphylococcus aureus and methicillin-resistant S. aureus. After employing filter paper disc method, the diameter of zone of bacterial inhibition (DZI) was measured as the parameter of antibacterial activity. The genomic DNA was extracted from all the tested plants and PCR amplified using trnH–psbA primer pair and subjected to DNA sequencing, followed by alignment analysis and dendrogram construction. Arogya and MKD did not show any antibacterial activity (DZI = 0.0 mm), whereas Sisila, BAN and MT showed antibacterial activity only against E. coli and S. aureus (mean DZI of 8.2 mm and 8.4 mm respectively). Bibila sweet and sour orange showed significantly higher antibacterial activity against all E. coli, S. aureus and methicillin-resistant S. aureus (mean DZI of 10.2 mm, 10.5 mm and 7.8 mm respectively). DNA barcoding provided unique sequence identifiers for each cultivar. These antibacterial activity data in combination with DNA barcodes could help to develop new cultivars through breeding to promote the sweet orange industry in Sri Lanka.The Journal of Agricultural Sciences, vol.11, No 1, January 2016, pp.13-23
Black pepper (Piper nigrum L.) is an important spice. The adulteration of black pepper seeds and powder with papaya seeds, green chili and red chili can be seen and limited studies have been conducted to detect these adulterants. The objectives of the present study were to assess the appropriateness of morphometric methods to discriminate papaya seed and chili adulterations in black pepper and to establish a DNA based strategy to detect these adulterations. A necessary adulteration series of seeds and powders were prepared for the analyses along with commercial samples. The appearance of the seed and powder samples were slightly different but not very distinct among the pure and adulterated samples emphasizing the need of a biochemical approach to detect the adulteration. The adulterated and commercial black pepper samples received lower pungency ranks compared to that of pure samples. QIAGEN DNeasy ® Plant Mini Kit was successful in extracting PCR amplifiable DNA from any sample without papaya seeds and the modified CTAB method was able to extract required PCR amenable DNA from any sample with papaya seed material. The universal DNA barcoding primer pair, psbA-trnH, was used to amplify the DNA. Black pepper DNA yielded 200 bp band, chili and papaya DNA yielded 450 bp band and DNA from adulterated samples produced both 200 bp and 450 bp bands. Therefore this strategy can be used to detect papaya / chili adulterations in black pepper.
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