3 research outputs found

    Identification and molecular characterisation of vascular-related defense genes against papaya dieback disease

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    Malaysia is one of the world’s main exporters of papaya with an export value worth RM120 million per year. An outbreak of papaya dieback disease on 2003 lowered the market value up to 60% from the previous year, and remains affected since. Erwinia mallotivora, the causal bacteria for papaya dieback disease, enters the papaya plant either through natural openings or wounds, to penetrate into the petioles and stem, and subsequently colonizes the entire vascular system. This results in the appearance of water-soak lesion at the infected region and disruption of the plant’s upper meristem region, which produces papaya fruits. To date, the only solution to this problem is by demolishing all the infected papaya plants as it can easily be transmitted to other plants nearby. An alternative approach such as genetic engineering to avoid mass destruction of matured papaya plants is deemed critical. Hence, the aims of this study were to identify potential vascular-related defense genes using bioinformatics approach, isolate and characterize the genes from Eksotika papaya and subsequently to assess the functionality of the genes in transformed papaya seedlings challenged with E. mallotivora. This study has successfully identified vascular-related defense genes against papaya dieback disease using bioinformatics approach. After data mining of bioinformatics databases including The Arabidopsis Information Resources (TAIR), Arabidopsis thaliana Trans-factor and cis-element Prediction Database (ATTED), Phytozome, and National Center for Biotechnology Information (NCBI), identified five potential genes were then mapped onto the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of Arabidopsis model plant. Orthologs of the Glycerol kinase (NHO1), Pathogenesis-related protein 1 (PR1b), Leucine-rich repeat receptor-like serine/threonine-protein kinase (EFR), RPM1 interacting protein 4 (RIN4) and Mitogen-activated protein kinase 4 (MPK4) genes were isolated and fully characterized from Carica papaya var. Eksotika I. NHO1 gene with an estimated size of 1572 base pair (bp) was hypothesized as defense-related gene against pathogen. The similar role was predicted with PR1b gene (492 bp), while EFR gene (741 bp) was postulated to be involved in pathogen signaling. In addition, RIN4 gene (540 bp) was suggested to be a member of resistance inducer upon pathogen invasion. MPK4 gene (1125 bp) was estimated to work in a cascade of mediating various responses against pathogens through different signaling pathway of defense responses. These genes were then sub cloned, in a sense orientation, into the pEAQ virus vector for subsequent transformation into papaya seedlings via Agrobacterium tumefaciens strain GV3101. The constructs were designated as pEAQ.NHO1, pEAQ.PR1, pEAQ.EFR, pEAQ.RIN4 and pEAQ.MPK4. Leaves of two months old papaya seedlings were infiltrated with A. tumefaciens strain GV3101 (OD600 = 1.5) harbouring each respective constructs. The transformed papaya seedlings were then challenged with E. mallotivora (cfu = 106) and exhibited disease symptoms development as early as day two after infection. Further profiling of the transgene expression in the papaya seedlings via real-time polymerase chain reaction (qRTPCR) analysis resulted in functional expression against the causal bacteria of this papaya dieback disease. Overexpression of PR1b and NHO1 genes resulted in high expression level in infected plants compared to the uninfected plants. While overexpression of MPK4 and RIN4 showed high fold changes compared to control plants, expression of MPK4 exhibited a repressed defense response via earlier development of symptoms in both infected and uninfected plants. In addition, overexpression of EFR showed a down-regulation expression for both infected and uninfected plants. These findings will lay the foundation for subsequent studies in developing a conceivable solution against this papaya dieback disease

    Development of Water Soluble NPK Fertilizer for Watermelon Cultivation Under Rain Shelter Structure

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    Commercial watermelon cultivation at open field depends on granular fertilizer for higher yield and quality. However, application of granular fertilizer is laborious, inefficient, inadequate nutrient uptake and can cause toxicity. Therefore, smart fertigation system as incorporated by IoT (Internet of things) appears to be the best option to improve such issues. To complete the smart fertigation system, application of water-soluble NPK fertilizer is the key tool. Therefore, an experiment was conducted to evaluate and select the optimum water soluble NPK rates used on growth, yield and fruit quality of watermelon under rain shelter structure. Watermelon seedlings were subjected to four water-soluble NPK fertilizer rates at 75%, 100% (Control - 108 kg N, 247.32 kg P and 153.6 kg K ), 125% and 150% based on common practices of NPK granular fertilizer used in watermelon at open field planting system for 65 days. Fertilizer rates induced at 150% significantly increased leaf number, leaf SPAD, total leaf area and leaf dry weight as compared to control at 14.17%, 16.22%, 21.55% and 15.06% respectively. Whereas, application of fertilizer at 125% significantly increased the leaf number, total leaf area and leaf dry weight with the respective increments of 11.5%, 17.19% and 13.08%. Based on regression trendline, application of 125% optimized the performances of the plants such as leaf number, total leaf area and fruit weight. To conclude, increasing 25% NPK standard fertilizer rate (125%) could be promising for watermelon cultivation throughout fertigation under open field planting system. Further trials in open field planting system needs to be undertaken to ensure the selected rates identified from this study could improve the whole performances of watermelo
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