Symptomatology and range of the blood disease bacterium A2 HR MARDI strain (Ralstonia syzygii subsp. celebensis) on selected hosts

Bacterial wilt disease is one of the major diseases in banana. In Malaysia, banana blood disease (BBD) is caused by the blood disease bacterium (BDB) A2 HR MARDI (Ralstonia syzygii subsp. celebensis). This disease bears similarities in symptomatology with Moko disease which caused by Ralstonia solanacearum and BBD in Indonesia, which caused by BDB R229. To determine the symptoms and host range of BDB, a pathogenicity test and host range study were carried out. In this study, there are four stages of external and internal symptoms which were observed. The pathogenicity of the bacterium cultures was then tested on banana, tomato and heliconia plantlets to determine the host range for BDB. To reconfirm that the banana was infected with BDB, re-isolation of BDB from the infected banana plants and Koch’s postulates test were performed. The results showed that there were symptoms of wilting and yellowing of leaves, which eventually caused plants death in the banana plantlets but no symptoms appeared in tomato and heliconia. The results indicate that BDB A2 HR MARDI is host-specific pathogen, only infecting banana similar to BDB R229 and is not as a broad range pathogen as R. solanacearum

Pengenalpastian dan pencirian gen trichoderma virens UKM1 mengekod enzim terlibat dalam pencuraian kitin krustasea

Kitin merupakan polisakarida struktur yang dapat dicurai oleh enzim kitinolisis kepada pelbagai terbitan yang boleh digunakan dalam bidang perubatan, pertanian dan rawatan air. Pengenalpastian dan pencirian gen-gen Trichoderma virens UKM1 mengekod enzim terlibat dalam pencuraian kitin krustasea telah dilakukan melalui penjanaan penanda jujukan terekspres (ESTs) dan analisis pengekspresan gen menggunakan mikroatur DNA. Sebanyak tiga perpustakaan cDNA T. virens UKM1 yang masing-masing diaruh oleh kitin, glukosamina dan kitosan telah dibina. Sejumlah 1536 klon cDNA telah dijujuk dan sebanyak 1033 ESTs berkualiti telah dijana. Seterusnya, perbezaan pengekspresan gen apabila pertumbuhan kulat diaruh dengan kehadiran kitin krustasea dan tanpa kitin pada hari ketiga dan kelima telah ditentukan. Sebanyak 1824 klon cDNA telah dititik ke atas slaid kaca dan dihibrid bersama dengan cDNA terlabel Cy3 atau Cy5 yang disintesis daripada mRNA yang dipencil daripada kulat yang ditumbuhkan dalam medium mengandungi kitin krustasea atau glukosa (kawalan). Sebanyak 91 dan 61 gen, masing-masing bagi hari ketiga dan kelima didapati terekspres melebihi dua gandaan apabila kulat menggunakan kitin krustasea sebagai sumber karbon. Beberapa gen mengekod kitinase seperti ech1 dan cht3 (endokitinase), nag1 (eksokitinase) dan nagB (glukosamina 6-P-deaminase) didapati terekspres dengan tinggi pada kedua-dua hari. Selain daripada itu, gen mengekod protein hidrofobin, protease serina dan beberapa protein hipotetik juga terekspres dengan tinggi dengan kehadiran kitin krustasea. Protein-protein ini dijangka memainkan peranan penting dalam membantu pencuraian kitin krustasea

Development of a Highly Sensitive Loop-Mediated Isothermal Amplification Incorporated with Flocculation of Carbon Particles for Rapid On-Site Diagnosis of Blood Disease Bacterium Banana

Bananas are one of the most crucial fruit crops worldwide and significantly contribute to food security in developing countries. However, blood disease of bananas caused by Ralstonia syzygii subspecies celebensensis has become a threat to banana production. Rapid and accurate diagnosis of BDB for on-site detection is pivotal at an early stage for an effective disease control strategy. This study developed LAMP with specific primers targeting BDB, followed by a flocculation assay for visualising positive amplification in the LAMP assay. The assay was sensitive to picogram amounts of gDNA (0.5 pg). LAMP assay on BDB gDNA showed flocculation, but negative results on Fusarium oxysporus cubense and Ralstonia solanacearum confirming the specificity of the assays. Field testing conducted at MARDI headquarters and Taman Pertanian Universiti discovered that the LAMP-flocculation assays were successful in detecting BDB on symptomatic samples as well as on samples from a healthy plot with no symptom observed at the sampling stage, revealing that this assay can detect BDB at an early infection stage. The validation results showed that the LAMP-flocculation assay was comparable with the PCR technique. This newly developed technique is highly specific and sensitive for the early detection of BDB for the adoption of precautionary control measures

Development of a Highly Sensitive Loop-Mediated Isothermal Amplification Incorporated with Flocculation of Carbon Particles for Rapid On-Site Diagnosis of Blood Disease Bacterium Banana

Bananas are one of the most crucial fruit crops worldwide and significantly contribute to food security in developing countries. However, blood disease of bananas caused by Ralstonia syzygii subspecies celebensensis has become a threat to banana production. Rapid and accurate diagnosis of BDB for on-site detection is pivotal at an early stage for an effective disease control strategy. This study developed LAMP with specific primers targeting BDB, followed by a flocculation assay for visualising positive amplification in the LAMP assay. The assay was sensitive to picogram amounts of gDNA (0.5 pg). LAMP assay on BDB gDNA showed flocculation, but negative results on Fusarium oxysporus cubense and Ralstonia solanacearum confirming the specificity of the assays. Field testing conducted at MARDI headquarters and Taman Pertanian Universiti discovered that the LAMP-flocculation assays were successful in detecting BDB on symptomatic samples as well as on samples from a healthy plot with no symptom observed at the sampling stage, revealing that this assay can detect BDB at an early infection stage. The validation results showed that the LAMP-flocculation assay was comparable with the PCR technique. This newly developed technique is highly specific and sensitive for the early detection of BDB for the adoption of precautionary control measures

Recombinant Protein Foliar Application Activates Systemic Acquired Resistance and Increases Tolerance Against Papaya Dieback Disease

Similar to animals, plants possess ‘immune memory’ in response to invading pathogens that lead to enhanced defense reaction following pathogen exposure. Systemic acquired resistance (SAR) is a well-characterized type of plant immunity and is associated with coordinated expression of a set of pathogenesis-related (PR) genes and proteins also known as SAR markers. Induction of SAR in plants was shown to be initiated by group of chemicals and biological compounds known as SAR inducers that can be used for the management of important plant diseases. Elucidation and characterization of potential SAR inducers as potential elicitors that can protect papaya from the papaya dieback disease pathogen were carried out using HRPX protein, which was produced as a recombinant protein in an Escherichia coli system. Disease severity analysis in a glasshouse experiment indicated lower disease infection rates in the HRPX-treated plants than in water-treated plants. Selected SAR-associated defense gene expression was also shown to increase in treated plants, via quantitative real-time PCR analysis, confirming enhanced disease response through SAR activation. In this report, the selected recombinant protein was shown to activate the SAR mechanism in papaya for increased tolerance against papaya dieback disease, which was proven via physiological and molecular analysis

Recombinant Protein Foliar Application Activates Systemic Acquired Resistance and Increases Tolerance Against Papaya Dieback Disease

Similar to animals, plants possess ‘immune memory’ in response to invading pathogens that lead to enhanced defense reaction following pathogen exposure. Systemic acquired resistance (SAR) is a well-characterized type of plant immunity and is associated with coordinated expression of a set of pathogenesis-related (PR) genes and proteins also known as SAR markers. Induction of SAR in plants was shown to be initiated by group of chemicals and biological compounds known as SAR inducers that can be used for the management of important plant diseases. Elucidation and characterization of potential SAR inducers as potential elicitors that can protect papaya from the papaya dieback disease pathogen were carried out using HRPX protein, which was produced as a recombinant protein in an Escherichia coli system. Disease severity analysis in a glasshouse experiment indicated lower disease infection rates in the HRPX-treated plants than in water-treated plants. Selected SAR-associated defense gene expression was also shown to increase in treated plants, via quantitative real-time PCR analysis, confirming enhanced disease response through SAR activation. In this report, the selected recombinant protein was shown to activate the SAR mechanism in papaya for increased tolerance against papaya dieback disease, which was proven via physiological and molecular analysis