In preliminary stage, the nucleotide sequence of Cs16 Colletotrichum gloeosporioides associated gene obtained from database of (NCBI), analyzed with Clustalx 1.83 in order to assess the deduced amino acid sequences. The full-length sequences of Cs16 gene is including with specific Open Reading Frame (ORF) having 3’ and 5’ UTR was obtained. In order to assess the relationship of C. gloeosporioides associated CS16 gene with other members, a combined rooted neighbor-joining (NJ) tree was generated through MEGA 4.5 by following 1000 bootstrap replicates. The resulted alignment predicts that Cs16 gene showed entirely diverse sequences from other species and has no any conserved domain sequences between them. In order to scrutinize the full length nucleotide sequences, the gene functional cloning of Cs16 gene has been carried out by PCR amplification. To examine the functional analysis of Cs16 gene in C. gloeosporioides and its regulatory mechanisms, the over expressing vector (pGapneoR26) was transformed into Agrobacterium strain LBA4404 that resulted successful detection of GFP-Cs16 expression into C. gloeosporioides transformants and it was also substantially detected by fluorescence microscopy. The T-DNA intentional mutants of C. gloeosporioides mycelia illustrated phonotypic different intensification, colour, colony growth rate than wild type isolated C. gloeosporioides STIJ6 strain.
Whitefly is one of the most widespread agricultural pests in the world. Essential oils might be used to control this insect in an environmentally responsible way. The fumigant, repellent, and anti-oviposition activity of ethanol-extracted essential oils of Trachyspermum ammi, Withania coagulans, and Murraya koenigii against Bemisia tabaci was investigated in this study. In the experiment, three essential oil concentrations (2.5 mg/mL, 5 mg/mL, and 10 mg/mL) were used. Trachyspermum ammi had the highest percentage of whitefly death in laboratory experiments due to its fumigant toxicity; the same tendency was found in contact toxicity and repellent effect. Mortality percent rises as the concentration of essential oil increases with bioassay time. As the concentration of essential oil grows with bioassay time, so does the mortality rate. The most adult whiteflies escaped from the treated plants' greenhouse due to the highest essential oil concentration. The greatest repellency was found with Trachyspermum ammi essential oil at 10 mg/mL. The essential oil had the greatest anti-oviposition efficacy against whiteflies. Trachyspermum ammi possessed the most potent anti-oviposition deterrent, followed by Withania coagulans in second place. Murraya koenigii finished third with moderate anti-oviposition, which affects the number of eggs produced in comparison to the control. As a consequence, these three oils might be used as an effective and environmentally acceptable bio-insecticide to control B. tabaci.
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