Search

Basal stem rot (BSR) of oil palm is a disastrous disease caused by a white-rot fungus Ganoderma boninense Pat. Non-ribosomal peptides (NRPs) synthesized by non-ribosomal peptide synthetases (NRPSs) are a group of secondary metabolites that act as fungal virulent factors during pathogenesis in the host. In this study, we aimed to isolate NRPS gene of G. boninense strain UPMGB001 and investigate the role of this gene during G. boninense-oil palm interaction. The isolated NRPS DNA fragment of 8322 bp was used to predict the putative peptide sequence of different domains and showed similarity with G. sinense (85%) at conserved motifs of three main NRPS domains. Phylogenetic analysis of NRPS peptide sequences demonstrated that NRPS of G. boninense belongs to the type VI siderophore family. The roots of 6-month-old oil palm seedlings were artificially inoculated for studying NRPS gene expression and disease severity in the greenhouse. The correlation between high disease severity (50%) and high expression (67-fold) of G. boninense NRPS gene at 4 months after inoculation and above indicated that this gene played a significant role in the advancement of BSR disease. Overall, these findings increase our knowledge on the gene structure of NRPS in G. boninense and its involvement in BSR pathogenesis as an effector gene., (© 2021. The Author(s).)

Ganoderma produces lignolytic enzymes that can degrade the lignin component of plant cell walls, causing basal stem rot to oil palms. Nitrogen sources may affect plant tolerance to root pathogens while hydrogen peroxide (H 2 O 2 ), salicylic acid (SA) and jasmonic acid (JA) play important roles in plant defense against pathogens. In this study, we examined the expression of genes encoding manganese peroxidase (MnP) and laccase (Lac) in Ganoderma boninense treated with different nitrogen sources (ammonium nitrate, ammonium sulphate, sodium nitrate and potassium nitrate), JA, SA and H 2 O 2 . Transcripts encoding MnP and Lac were cloned from G. boninense . Of the three GbMnP genes, GbMnP_U6011 was up-regulated by all nitrogen sources examined and H 2 O 2 but was down-regulated by JA. The expression of GbMnP_U87 was only up-regulated by JA while GbMnP_35959 was up-regulated by ammonium nitrate but suppressed by sodium nitrate and down-regulated by H 2 O 2 . Among the three GbLac genes examined, GbLac_U90667 was up-regulated by ammonium nitrate, JA, SA and H 2 O 2 ; GbLac_U36023 was up-regulated by JA and H 2 O 2 while GbLac_U30636 was up-regulated by SA but suppressed by ammonium sulphate, sodium nitrate, JA and H 2 O 2 . Differential expression of these genes may be required by their different functional roles in G. boninense .

Plant defensins are plant defence peptides that have many different biological activities, including antifungal, antimicrobial, and insecticidal activities. A cDNA (EgDFS) encoding defensin was isolated from Elaeis guineensis. The open reading frame of EgDFS contained 231 nucleotides encoding a 71-amino acid protein with a predicted molecular weight at 8.69 kDa, and a potential signal peptide. The eight highly conserved cysteine sites in plant defensins were also conserved in EgDFS. The EgDFS sequence lacking 30 amino acid residues at its N-terminus (EgDFSm) was cloned into Escherichia coli BL21 (DE3) pLysS and successfully expressed as a soluble recombinant protein. The recombinant EgDFSm was found to be a thermal stable peptide which demonstrated inhibitory activity against the growth of G. boninense possibly by inhibiting starch assimilation. The role of EgDFSm in oil palm defence system against the infection of pathogen G. boninense was discussed.

Nitric oxide associated 1 (NOA1) protein is implicated in plant disease resistance and nitric oxide (NO) biosynthesis. A full-length cDNA encoding of NOA1 protein from oil palm (Elaeis guineensis) was isolated and designated as EgNOA1. Sequence analysis suggested that EgNOA1 was a circular permutated GTPase with high similarity to the bacterial YqeH protein of the YawG/YlqF family. The gene expression of EgNOA1 and NO production in oil palm root tissues treated with Ganoderma boninense, the causal agent of basal stem rot (BSR) disease were profiled to investigate the involvement of EgNOA1 during fungal infection and association with NO biosynthesis. Real-time PCR (qPCR) analysis revealed that the transcript abundance of EgNOA1 in root tissues was increased by G. boninense treatment. NO burst in Ganoderma-treated root tissue was detected using Griess reagent, in advance of the up-regulation of the EgNOA1 transcript. This indicates that NO production was independent of EgNOA1. However, the induced expression of EgNOA1 in Ganoderma-treated root tissues implies that it might be involved in plant defense responses against pathogen infection., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

Glucanases are enzymes that hydrolyze a variety β-d-glucosidic linkages. Plant β-1,3-glucanases are able to degrade fungal cell walls; and promote the release of cell-wall derived fungal elicitors. In this study, three full-length cDNA sequences encoding oil palm (Elaeis guineensis) glucanases were analyzed. Sequence analyses of the cDNA sequences suggested that EgGlc1-1 is a putative β-d-glucan exohydolase belonging to glycosyl hydrolase (GH) family 3 while EgGlc5-1 and EgGlc5-2 are putative glucan endo-1,3-β-glucosidases belonging to GH family 17. The transcript abundance of these genes in the roots and leaves of oil palm seedlings treated with Ganoderma boninense and Trichoderma harzianum was profiled to investigate the involvement of these glucanases in oil palm during fungal infection. The gene expression of EgGlc1-1 in the root of oil palm seedlings was increased by T. harzianum but suppressed by G. boninense; while the gene expression of both EgGlc5-1 and EgGlc5-2 in the roots of oil palm seedlings was suppressed by G. boninense or/and T. harzianum., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

Background: Basal stem rot (BSR) disease caused by the fungus Ganoderma boninense is the most serious disease affecting the oil palm; this is because the disease escapes the early disease detection. The biocontrol agent Trichoderma harzianum can protect the disease only at the early stage of the disease. In the present study, the expression levels of three oil palm (Elaeis guineensis Jacq.) chitinases encoding EgCHI1, EgCHI2, and EgCHI3 at 2, 5, and 8 weeks inoculation were measured in oil palm leaves from plants treated with G. boninense or T. harzianum alone or both., Methods: The five-month-old oil palm seedlings were treated with Gano-wood blocks inoculum and trichomulch. Expression of EgCHI1, EgCHI2, and EgCHI3 in treated leaves tissue was determined by real-time PCR., Results: Oil palm chitinases were not strongly expressed in oil palm leaves of plants treated with G. boninense alone compared to other treatments. Throughout the 8-week experiment, expression of EgCHI1 increased more than 3-fold in leaves of plants treated with T. harzianum and G. boninense when compared to those of control and other treated plants., Conclusion: The data illustrated that chitinase cDNA expression varied depending on tissue and the type of treatment.

Aims: This study was designed to examine the enzyme activity of selected virulent isolates of Ganoderma boninense against oil palm. In a separate in vitro assessment, the effect of macronutrients on the mycelial growth of four selected Ganoderma spp. was also tested. Methodology and results: The study involved a comparison of ligninolytic enzymes; lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase (Lac) profiling of eight isolates of G. boninense, categorized into three levels of aggressiveness, with two control isolates (G. boninense PER71 and G. tornatum NPG1) using solid-state fermentation (SSF). The Principal Component Analysis (PCA) revealed that the isolates had a significant production of ligninolytic enzymes on day 80. The most aggressive isolate, ET61 had the highest Lac production. As for the macronutrient test, mycelial growth for all the Ganoderma spp. was highly affected by potassium (K). Conclusion, significance and impact of study: The findings of this study elucidated the characteristics of G. boninense in relation to enzyme production for the degradation of oil palm lignin and the identification of essential nutrients involved in the survival and growth of Ganoderma spp. The study provides vital information on the pathogenic characteristics of G. boninense isolates involved in biomass degradation along with the role of nutrient on the growth of Ganoderma spp. that may influence basal stem rot (BSR) management in the field. [ABSTRACT FROM AUTHOR]

Cerato-platanin (CP) proteins are small extracellular proteins that play essential roles in the plant-pathogen interactions of some economically important filamentous fungi. At present, there is a lack of information about the roles of CP proteins secreted by Ganoderma boninense , the causal agent of basal stem rot disease in African oil palm, during the early host-pathogen interaction. This work studies the properties and roles of two CP proteins from G. boninense strain PER71. Using bioassay-guided fractionation and nano-LC-MS/MS analysis, we identified peptide matches for G. sinense ZZ0214-1 CP-like and other elicitor proteins in the culture filtrate of G. boninense strain PER71. We found that GbCP2 and GbCP6, two out of nine CP gene models identified in the fungal genome of G. boninense strain PER71, exhibited potential effector properties. We further validated their effector properties by isolating and characterising their coding regions. Interestingly, GbCP6 showed a significant expression level at 11 days post inoculation, indicating its significance in fungal pathogenesis. On the other hand, GbCP2 did not show differential expression at any sampling point, suggesting a different function or role in the pathogenicity of G. boninense. This study identifies GbCP6 as a potential biomarker for early detection of basal stem rot disease in oil palm. [Display omitted] • Ganoderma boninense secretes hypersensitive response-inducing elicitor proteins. • Nine cerato-platanin homologs are present in the G. boninense fungal genome. • GbCP2 and GbCP6 are effector proteins with potential implications in pathogenesis. • GbCP6 emerges as a potential early biomarker for basal stem rot disease. [ABSTRACT FROM AUTHOR]

Basal stem rot (BSR) is the most devastating disease of oil palm. In this study, we examined the transcriptional responses of oil palm roots treated with a causal agent of BSR, Ganoderma boninense using a cDNA microarray approach. A total of 61 from 3,748 transcripts examined were found to be significantly up- or down-regulated in oil palm roots infected with G. boninense at 3 and 6 weeks post inoculation compared to those from uninfected roots. The differentially expressed genes identified in the artificially infected oil palm roots included genes encoding isoflavone reductase, Em protein H2, SPX domain-containing protein 1, pathogenesis-related protein 1, vicilin-like antimicrobial peptide. The gene expression of isoflavone reductase, which is involved in the production of phytoalexin and three related genes in the phenylpropanoid biosynthetic pathway was also profiled in the treated oil palms using real-time quantitative reverse transcription PCR. This information has contributed to our understanding of the defense mechanisms of oil palm in response to G. boninense, the future development of molecular markers for marker assisted breeding and screening of oil palms that are tolerant to G. boninense. [ABSTRACT FROM AUTHOR]

Fungal necrosis and ethylene inducing proteins (NEPs) induce cell death and elicit strong immune responses in many crops. In this study, we report the cloning and characterization of a transcript encoding NEP from Ganoderma boninense which belongs to a family of fungi in Ganodermataceae that cause serious infections of cacao, rubber, tea, coffee and palms. The open reading frame (ORF) encoding NEP in G. boninense (GbNEP) was cloned by 5′ and 3' rapid amplification of cDNA ends (RACE) PCR. The transcript abundance of GbNEP increased in fungal culture treated with salicylic acid and jasmonic acid, respectively. The soluble recombinant GbNEP expressed in Escherichia coli BL21(DE3)pLysS was able to induce necrosis in tobacco and tomato but ineffective when applied to oil palm leaves and root tissues. The recombinant GbNEP could induce localized cell death, and production of hydrogen peroxide and superoxide in tobacco leaves. The addition of LaCl 3 (a calcium channel inhibitor) and EGTA (a Ca2+ chelator) to tobacco leaves prior to the challenge with GbNEP significantly reduced the necrosis symptoms indicating that Ca2+ is required for the action of GbNEP. In summary, this is the first report of NEP from Ganoderma species which may contribute to further research on the role of GbNEP during disease development. • First report of necrosis and ethylene inducing protein (NEP) from Ganoderma boninense. • The transcript abundance of GbNEP increased in G. boninense culture treated with salicylic acid and jasmonic acid, respectively. • Soluble recombinant GbNEP was able to induce necrosis in tobacco and tomato but ineffective when applied to oil palm leaves and root tissues. • Recombinant GbNEP could induce defense responses in tobacco leaves. • Calcium is required for the action of GbNEP. [ABSTRACT FROM AUTHOR]

Oil palm is subjected to various diseases including Basal Stem Rot (BSR) which is mainly caused by the basidiomycetes fungi, Ganoderma. However, studies on the fungal infection mechanism especially at molecular level and the biological processes involved are still very limited. Fungal cyclophilin (CYP) has been reported to be involved in fungal pathogenicity but involvement of CYP in the pathogenicity of Ganoderma boninense has not been revealed yet. The main objective of this study is to isolate cDNAs encoding CYP and to profile the expression level of these transcripts during infection stages of G. boninense. In this study, five full-length cDNAs encoding CYP were successfully isolated from G. boninense . They were classified as different family members of CYP because significant differences could be observed on their coding, 5′ or 3′ un-translated regions (UTRs). The expression of these five CYP transcripts in different type of G. boninense tissues and the infecting mycelia tissues was studied using real time quantitative PCR (qPCR). Based on the expression results and analysis, the potential functions of these CYP transcripts were predicted to be involved in the development of fruiting body (GbCYP201) and fungal stress response or pathogenicity (GbCYP203 and GbCYP205). [ABSTRACT FROM AUTHOR]

Abstract: Basal stem rot (BSR) is a major disease of oil palm caused by a pathogenic fungus, Ganoderma boninense. However, the interaction between the host plant and its pathogen is not well characterized. To better understand the response of oil palm to G. boninense, transcript profiles of eleven putative defence-related genes from oil palm were measured by quantitative reverse-transcription (qRT)-PCR in the roots of oil palms treated with G. boninense from 3 to 12 weeks post infection (wpi). These transcripts encode putative Bowman–Birk serine protease inhibitors (EgBBI1 and 2), defensin (EgDFS), dehydrin (EgDHN), early methionine-labeled polypeptides (EgEMLP1 and 2), glycine-rich RNA binding protein (EgGRRBP), isoflavone reductase (EgIFR), metallothionein-like protein (EgMT), pathogenesis-related-1 protein (EgPRP), and type 2 ribosome-inactivating protein (EgT2RIP). The transcript abundance of EgBBI2 increased in G. boninense-treated roots at 3 and 6wpi compared to those of controls; while the transcript abundance of EgBBI1, EgDFS, EgEMLP1, EgMT, and EgT2RIP increased in G. boninense-treated roots at 6 or 12wpi. Meanwhile, the gene expression of EgDHN was up-regulated at all three time points in G. boninense-treated roots. The expression profiles of the eleven transcripts were also studied in leaf samples upon inoculation of G. boninense and Trichoderma harzianum to identify potential biomarkers for early detection of BSR. Two candidate genes (EgEMLP1 and EgMT) that have different profiles in G. boninense-treated leaves compared to those infected by T. harzianum may have the potential to be developed as biomarkers for early detection of G. boninense infection. [Copyright &y& Elsevier]