Bioprospecting Study of Plant Growth Promoting Rhizospheric Bacteria from Oil Palm Plantation as Biological Control Agent of Ganoderma boninense.

<b>Background and Objective:</b> The prioritisation of oil palm studies involves the exploration of novel bacterial isolates as possible agents for suppressing <i>Ganoderma boninense</i>. The objective of this study was to evaluate and characterise the potential of rhizospheric bacteria, obtained from the rhizosphere of oil palm plants, in terms of their ability to demonstrate anti-<i>Ganoderma </i>activity. <b>Materials and Methods:</b> The study began by employing a dual culture technique to select hostile bacteria. Qualitative detection was performed to assess the antifungal activity, as well as the synthesis of chitinase and glucanase, from certain isolates. The candidate strains were molecularly identified using 16S-rRNA ribosomal primers, specifically the 27F and 1492R primers. <b>Results:</b> The findings of the study indicated that the governmental plantation exhibited the highest ratio between diazotroph and indigenous bacterial populations in comparison to the other sites. Out of a pool of ninety bacterial isolates, a subset of twenty-one isolates demonstrated the ability to impede the development of <i>G. boninense</i>, as determined using a dual culture experiment. Twenty-one bacterial strains were found to exhibit antifungal activity. Nine possible bacteria were found based on the sequence analysis. These bacteria include <i>Burkholderia territorii</i> (RK2, RP2, RP3, RP5), <i>Burkholderia stagnalis</i> (RK3), <i>Burkholderia cenocepacia</i> (RP1), <i>Serratia marcescens</i> (RP13) and <i>Rhizobium multihospitium</i> (RU4). <b>Conclusion:</b> The findings of the study revealed that a significant proportion of the bacterial population exhibited the ability to perform nitrogen fixation, indole-3-acetic acid (IAA) production and phosphate solubilization. However, it is worth noting that <i>Rhizobium multihospitium</i> RU4 did not demonstrate the capacity for phosphate solubilization, while <i>B. territory</i> RK2 did not exhibit IAA production.

Whole-genome sequencing of Ganoderma boninense, the causal agent of basal stem rot disease in oil palm, via combined short- and long-read sequencing.

The hemibiotrophic Basidiomycete pathogen Ganoderma boninense (Gb) is the dominant causal agent of oil palm basal stem rot disease. Here, we report a complete chromosomal genome map of Gb using a combination of short-read Illumina and long-read Pacific Biosciences (PacBio) sequencing platforms combined with chromatin conformation capture data from the Chicago and Hi-C platforms. The genome was 55.87 Mb in length and assembled to a high contiguity (N50: 304.34 kb) of 12 chromosomes built from 112 scaffolds, with a total of only 4.34 Mb (~ 7.77%) remaining unplaced. The final assemblies were evaluated for completeness of the genome by using Benchmarking Universal Single Copy Orthologs (BUSCO) v4.1.4, and based on 4464 total BUSCO polyporales group searches, the assemblies yielded 4264 (95.52%) of the conserved orthologs as complete and only a few fragmented BUSCO of 42 (0.94%) as well as a missing BUSCO of 158 (3.53%). Genome annotation predicted a total of 21,074 coding genes, with a GC content ratio of 59.2%. The genome features were analyzed with different databases, which revealed 2471 Gene Ontology/GO (11.72%), 5418 KEGG (Kyoto Encyclopedia of Genes and Genomes) Orthologous/KO (25.71%), 13,913 Cluster of Orthologous Groups of proteins/COG (66.02%), 60 ABC transporter (0.28%), 1049 Carbohydrate-Active Enzymes/CAZy (4.98%), 4005 pathogen-host interactions/PHI (19%), and 515 fungal transcription factor/FTFD (2.44%) genes. The results obtained in this study provide deep insight for further studies in the future.

Fumaric acid production by Rhizopus species from acid hydrolysate of oil palm empty fruit bunches.

The hemicellulosic fraction of lignocellulosic biomass is a very important material, due to the significant concentration of pentoses present in its composition and that can be used sustainably in biotechnological processes such as the production of fumaric acid. Research efforts are currently being promoted for the proper disposal and valorization of empty fruit bunches (EFB) from oil palm. In this work, seventeen Rhizopus species were evaluated in a fermentation medium with EFB hydrolyzate, without detoxification, as a carbon source for fumaric acid production. Rhizopus circicans 1475 and Rhizopus 3271 achieved productions of 5.65 g.L-1 and 5.25 g.L-1 of fumaric acid at 30 °C, 120 rpm for 96 h, respectively. The percentage of consumed sugars, mainly pentoses, was 24.88% and 34.02% for R. circicans 1475 and R 3271, respectively. Soy peptone and ammonium sulfate were evaluated as nitrogen sources, where soy peptone stimulated the formation of biomass pellets while ammonium sulfate produced mycelia and clamps.

Fumaric acid production from fermented oil palm empty fruit bunches using fungal isolate K20: a comparison between free and immobilized cells.

This study investigated the potential of using steam-exploded oil palm empty fruit bunches (EFB) as a renewable feedstock for producing fumaric acid (FA), a food additive widely used for flavor and preservation, through a separate hydrolysis and fermentation process using the fungal isolate K20. The efficiency of FA production by free and immobilized cells was compared. The maximum FA concentration (3.25 g/L), with 0.034 g/L/h productivity, was observed after incubation with the free cells for 96 h. Furthermore, the production was scaled up in a 3-L air-lift fermenter using oil palm EFB-derived glucose as the substrate. The FA concentration, yield, and productivity from 100 g/L initial oil palm EFB-derived glucose were 44 g/L, 0.39 g/g, and 0.41 g/L/h, respectively. The potential for scaling up the fermentation process indicates favorable results, which could have significant implications for industrial applications.

Comparison of Ganoderma boninense Isolate's Aggressiveness Using Infected Oil Palm Seedlings.

Basal stem rot incidence caused by a white-rot fungus, Ganoderma boninense, is the major disease of oil palm in Southeast Asia. The rate of disease transmission and host damage are affected by variations in pathogen aggressiveness. Several other studies have used the disease severity index (DSI) to determine G. boninense aggressiveness levels while verifying disease using a culture-based method, which might not provide accurate results or be feasible in all cases. To differentiate G. boninense aggressiveness, we employed the DSI and vegetative growth measurement of infected oil palm seedlings. Disease confirmation was performed through scanning electron microscopy and molecular identification of fungal DNA from both infected tissue and fungi isolated from Ganoderma selective medium. Two-month-old oil palm seedlings were artificially inoculated with G. boninense isolates (2, 4A, 5A, 5B, and 7A) sampled from Miri (Lambir) and Mukah (Sungai Meris and Sungai Liuk), Sarawak. The isolates were categorized into three groups: highly aggressive (4A and 5B), moderately aggressive (5A and 7A), and less aggressive (2). Isolate 5B was identified as the most aggressive, and it was the only one to result in seedling mortality. Out of the five vegetative growth parameters measured, only the bole size between treatments was not affected. The integration of both conventional and molecular approaches in disease confirmation allows for precise detection.

Evidence for high gene flow, nonrandom mating, and genetic bottlenecks of Ganoderma boninense infecting oil palm (Elaeis guineensis Jacq.) plantations in Malaysia and Indonesia.

Ganoderma boninense, the causal agent of basal stem rot (BSR) disease, has been recognized as a major economic threat to commercial plantings of oil palm (Elaeis guineensis Jacq.) in Southeast Asia, which supplies 86% of the world's palm oil. High genetic diversity and gene flow among regional populations of 417 G. boninense isolates collected from Sabah, Sarawak, and Peninsular Malaysia (Malaysia) and Sumatra (Indonesia) were demonstrated using 16 microsatellite loci. Three genetic clusters and different admixed populations of G. boninense across regions were detected, and they appeared to follow the spread of the fungus from the oldest (Peninsular Malaysia and Sumatra) to younger generations of oil palm plantings (Sabah and Sarawak). Low spatial genetic differentiation of G. boninense (FST = 0.05) among the sampling regions revealed geographically nonrestricted gene dispersal, but isolation by distance was still evident. Analysis of molecular variance (AMOVA) confirmed the little to no genetic differentiation among the pathogen populations and the three genetic clusters defined by STRUCTURE and minimum spanning network. Despite G. boninense being highly outcrossing and spread by sexual spores, linkage disequilibrium was detected in 7 of the 14 populations. Linkage disequilibrium indicated that the reproduction of the fungus was not entirely by random mating and genetic drift could be an important structuring factor. Furthermore, evidence of population bottleneck was indicated in the oldest oil palm plantations as detected in genetic clusters 2 and 3, which consisted mainly of Peninsular Malaysia and Sumatra isolates. The population bottleneck or founding event could have arisen from either new planting or replanting after the removal of large number of palm hosts. The present study also demonstrated that migration and nonrandom mating of G. boninense could be important for survival and adaptation to new palm hosts.

Estudo de métodos naturais para redução de doenças fúngicas em sementes nativas da região Amazônica / Study of natural methods for reducing fungal diseases in native seeds in the Amazon region

Existe uma preocupação na produção de alimentos com a segurança sanitária desde o processo de cultivo até a sua expedição aos centros comerciais. Uma das maiores contaminações em sementes por suas características físicas e químicas, é a contaminação fúngica devida à acidez e umidade dos frutos. As doenças fúngicas entre elas, antracnose, vassoura de bruxa e podridão parda são consideradas importante doenças em pós colheita, ocorrendo, principalmente, sob condições de alta umidade relativa do ar e temperaturas elevadas (26ºC a 28ºC). Dentre as sementes da região Amazônica, destacam-se sementes de (cacau- Theobroma cacao L., pupunha- Bactris gasipaes, tucumã da Amazonas-Astrocaryum aculeatum), que apresentam problemas de contaminação fúngica durante o sistema de cultivo. Métodos de biocontroles com ação antimicrobiana: bactérias endofíticas, rizobactérias e fungo Trichoderma ssp, entre outros estão sendo estudados para a redução destas contaminações fúngicas em estudos in vitro e no campo. O objetivo deste trabalho é apresentar uma revisão bibliográfica sobre a utilização de métodos de biocontrole como uma alternativa promissora no manejo de doenças de plantas na fase de pós-colheita, com excelentes resultados em culturas de grande importância econômica. Assim novas alternativas ecologicamente sustentáveis demonstram a possibilidade de os produtos estudados serem utilizados no manejo da antracnose na pós-colheita.(AU)

There is a concern in food production with health security from cultivation process to its dispatch to redistribution centers. One of the biggest contaminations in seeds due to their physical and chemical characteristics, is the fungal contamination due to the acidity and humidity of the fruits. Fungal diseases including anthracnose, bruca's broom and brownrot are considered important diseases in post-harvest, occurring mainly under conditions of high relative humidity and high temperatures (26ºC to 28ºC). Among the seeds of the Amazon region, seeds of de (cacau- Theobroma cacao L., pupunha- Bactris gasipaes, tucumã da Amazonas- Astrocaryum aculeatum ), that present problems fungals contamination during in their growing cultures sistems. Biocontrol methods with antimicrobial action: endophytic bacteria, rhizobacteria and the fungus Trichoderma ssp, among others, are being studied to reduce these fungal contaminations in in vitro and field studies. The objective of this work is to present a bibliographic review on the use of biocontrol methods as an alternative that is promising in the management of plant diseases in the post-harvest phase, with excellent results in crops of great economic importance. Thus, new ecologically sustainable alternatives demonstrate the possibility of the studied products to be used in the management of anthracnose in the post-harvest period.(AU)

Antifungal Peptides from a Burkholderia Strain Suppress Basal Stem Rot Disease of Oil Palm.

Basal stem rot (BSR) is the most common disease of oil palm (Elaeis guineensis) in Southeast Asia. BSR is caused by a white-rot fungus Ganoderma boninense. The disease is difficult to manage. Therefore, development of novel and environmentally safe approaches to control the disease is important. Species of Burkholderia are known to have diverse lifestyles, some of which can benefit plants by suppressing diseases or increasing plant growth. In the present study, antifungal peptides produced by a bacterial strain isolated from the rhizosphere of an oil palm tree, Burkholderia sp. strain CP01, exhibited strong growth inhibition on G. boninense. A loss-of-function mutant of CP01 was generated, and it has enabled the identification of a 1.2-kDa peptide and its variants as the active antifungal compounds. High-resolution mass spectrometry revealed six analogous compounds with monoisotopic masses similar to the previously reported cyclic lipopeptides occidiofungin and burkholdine. The antifungal compounds of CP01 were secreted into media, and we sought to use CP01 culture extract without living cells to control BSR disease. Glasshouse experiments showed that CP01 culture extract suppressed BSR disease in oil palm seedlings. The ability of CP01 to produce an antifungal substance and suppress plant disease suggests its potential applications as a biofungicide in agriculture.

Phenazine from Pseudomonas aeruginosa UPMP3 induced the host resistance in oil palm (Elaeis guineensis Jacq.)-Ganoderma boninense pathosystem.

Pseudomonas aeruginosa developed its biocontrol agent property through the production of antifungal derivatives, with the phenazine among them. In this study, the applications of crude phenazine synthesized by Pseudomonas aeruginosa UPMP3 and hexaconazole were comparatively evaluated for their effectiveness to suppress basal stem rot infection in artificially G. boninense-challenged oil palm seedlings. A glasshouse experiment under the randomized completely block design was set with the following treatments: non-inoculated seedlings, G. boninense inoculated seedlings, G. boninense inoculated seedlings with 1 mg/ml phenazine application, G. boninense inoculated seedlings with 2 mg/ml phenazine application and G. boninense inoculated seedlings with 0.048 mg/ml hexaconazole application. Seedlings were screened for disease parameters and plant vigour traits (plant height, plant fresh weight, root fresh, and dry weight, stem diameter, and total chlorophyll) at 1-to-4 month post-inoculation (mpi). The application of 2 mg/ml phenazine significantly reduced disease severity (DS) at 44% in comparison to fungicide application (DS = 67%). Plant vigour improved from 1 to 4 mpi and the rate of disease reduction in seedlings with phenazine application (2 mg/ml) was twofold greater than hexaconazole. At 4, 6 and 8 wpi, an up-regulation of chitinase and ß-1,3 glucanase genes in seedlings treated with phenazine suggests the involvement of induced resistance in G. boninense-oil palm pathosystem.

GanoCare® Improves Oil Palm Growth and Resistance against Ganoderma Basal Stem Rot Disease in Nursery and Field Trials.

Basal stem rot (BSR) caused by Ganoderma boninense is a major threat to sustainable oil palm production especially in Southeast Asia and has brought economic losses to the oil palm industry around the world. With no definitive cure at present, this study introduces a new fertilizer technology called GanoCare®, as an effort to suppress BSR incidence in oil palm. Experiments were carried out to evaluate the effect of GanoCare® on growth, physiology, and BSR disease suppression using sitting technique in the oil palm nursery stage. A follow-up using similar treatments was carried out in the field to test on severity of Ganoderma using baiting technique under natural condition. Treatments tested were 10 g/month and 30 g/three months given as pretreatment only or continuous treatment. Results showed that GanoCare® increased the height, bulb diameter, leaf area, chlorophyll content, photosynthesis rate, and fresh and dry weight of the leaf, bole, and root of oil palm seedlings in the nursery trial. Seedlings treated with GanoCare® exhibited reduced percentage of disease severity, incidence, and dead seedlings, compared to the control. In nursery and field, lowest percentage of dead seedlings due to Ganoderma was found in seedlings given combination of pretreatment and continuous treatment of 30 g/three months (T4) with 5.56 and 6.67%, while control seedlings significantly marked the maximum percentage of 94.45 and 93.33%. The most successful treatment in both nursery and field was T4 with disease reductions of 77.78 and 82.36%, respectively, proving that nutrients contained in GanoCare® are essential in allowing better development of a strong defense system in the seedlings.

Application of Ground-Based LiDAR for Analysing Oil Palm Canopy Properties on the Occurrence of Basal Stem Rot (BSR) Disease.

Ground-based LiDAR also known as Terrestrial Laser Scanning (TLS) technology is an active remote sensing imaging method said to be one of the latest advances and innovations for plant phenotyping. Basal Stem Rot (BSR) is the most destructive disease of oil palm in Malaysia that is caused by white-rot fungus Ganoderma boninense, the symptoms of which include flattening and hanging-down of the canopy, shorter leaves, wilting green fronds and smaller crown size. Therefore, until now there is no critical investigation on the characterisation of canopy architecture related to this disease using TLS method was carried out. This study proposed a novel technique of BSR classification at the oil palm canopy analysis using the point clouds data taken from the TLS. A total of 40 samples of oil palm trees at the age of nine-years-old were selected and 10 trees for each health level were randomly taken from the same plot. The trees were categorised into four health levels - T0, T1, T2 and T3, which represents the healthy, mildly infected, moderately infected and severely infected, respectively. The TLS scanner was mounted at a height of 1 m and each palm was scanned at four scan positions around the tree to get a full 3D image. Five parameters were analysed: S200 (canopy strata at 200 cm from the top), S850 (canopy strata at 850 cm from the top), crown pixel (number of pixels inside the crown), frond angle (degree of angle between fronds) and frond number. The results taken from statistical analysis revealed that frond number was the best single parameter to detect BSR disease as early as T1. In classification models, a linear model with a combination of parameters, ABD - A (frond number), B (frond angle) and D (S200), delivered the highest average accuracy for classification of healthy-unhealthy trees with an accuracy of 86.67 per cent. It also can classify the four severity levels of infection with an accuracy of 80 per cent. This model performed better when compared to the severity classification using frond number. The novelty of this research is therefore on the development of new approach to detect and classify BSR using point clouds data of TLS.

Chitinasiproducens palmae gen. nov., sp. nov., a new member of the family Burkholderiaceae isolated from leaf tissues of oil palm (Elaeis guineensis Jacq.).

A Gram-stain-negative, aerobic, rod-shaped, leaf-associated bacterium, designated JS23T, was isolated from surface-sterilized leaf tissue of an oil palm grown in Singapore and was investigated by polyphasic taxonomy. Phylogenetic analyses based on 16S rRNA gene sequences and 180 conserved genes in the genome of several members of Burkholderiaceae revealed that strain JS23T formed a distinct evolutionary lineage independent of other taxa within the family Burkholderiaceae. The predominant ubiquinone was Q-8. The primary polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, and an unidentified aminophospholipid. The major fatty acids were C16 : 0, summed feature 3 (C16 : 1 ω7c /C16 : 1 ω6c) and summed feature 8 (C18 : 1 ω7c /C18 : 1 ω6c). The size of the genome is 5.36 Mbp with a DNA G+C content of 66.2 mol%. Genomic relatedness measurements such as average nucleotide identity, genome-to-genome distance and digital DNA-DNA hybridization clearly distinguished strain JS23T from the closely related genera Burkholderia, Caballeronia, Mycetohabitans, Mycoavidus, Pandoraea, Paraburkholderia, Robbsia and Trinickia. Furthermore, average amino acid identity values and the percentages of conserved proteins, 56.0-68.4 and 28.2-45.5, respectively, were well below threshold values for genus delineation and supported the assignment of JS23T to a novel genus. On the basis of the phylogenetic, biochemical, chemotaxonomic and phylogenomic evidence, strain JS23T is proposed to represent a novel species of a new genus within the family Burkholderiaceae, for which the name Chitinasiproducens palmae gen. nov., sp. nov., is proposed with the type strain of JS23T (= DSM 27307T=KACC 17592T).

Phytobacter palmae sp. nov., a novel endophytic, N2 fixing, plant growth promoting Gammaproteobacterium isolated from oil palm (Elaeis guineensis Jacq.).

A novel strain S29T with high nitrogen fixing ability was isolated from surface-sterilized leaf tissues of oil palm (Elaeis guineensis) growing in Science Park II, Singapore. On the basis of 16S rRNA gene analysis and multilocus sequence typing with the rpoB, gyrB, infB and atpD genes, strain S29T was a member of the genus Phytobacter, with Phytobacter ursingii ATCC 27989T and Phytobacter diazotrophicus LS 8T as its closest relatives. Unique biochemical features that differentiated strain S29T from its closest relatives were the ability to utilize melibiose, α-cyclodextrin, glycogen, adonital, d-arabitol, m-inositol and xylitol. The major fatty acids were C14 : 0, C16 : 0, C17 : 0, C16 : 1 ω5c and summed feature 2 containing C16 : 1 ω7c and/or C16 : 1 ω6c. The polar fatty acid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unidentified aminophospholipid and aminolipids. The draft genome of strain S29T comprised 5, 284, 330 bp with a G + C content of 52.6 %. The average nucleotide identity and digital DNA-DNA hybridization values between strain S29T and the phylogenetically related Enterobacterales species were lower than 95 % and 70 %, respectively. Thus, the polyphasic evidences generated through the phenotypic, chemotaxonomic and genomic methods confirmed that strain S29T represents a novel species of the genus Phytobacter, for which we propose the name Phytobacter palmae sp. nov. with the type strain of S29T (=DSM 27342T=KACC 17598T).

First report of the oil palm disease fungus Marasmius palmivorus from Taiwan causing stem rot disease on native Formosa palm Arenga engleri as new host.

Basidiomata were found on dead stems of wild native Formosa palm (Arenga engleri) in Taiwan. The fungus was identified based on morphology and internal transcribed spacer sequence comparison as the oil palm pathogen Marasmius palmivorus. A pathogenicity test with cultivated mycelium of M. palmivorus positively produced disease symptoms and death of non-wounded Formosa palm tree seedlings under excessive moisture conditions. These results indicate that mycelial inoculum may be more important for pathogenesis than spore inoculum and that the fungus does not require wounds for entry into the plant. Host records in the literature are critically revised. The extended geographical and host distribution indicate a greater risk by M. palmivorus in palm plantations than hitherto anticipated. SIGNIFICANCE AND IMPACT OF THE STUDY: Certain palm species are susceptible to disease caused by Marasmius palmivorus, particularly in oil and coconut palm plantations in tropical countries. Hitherto, there is no published information on the morphology and pathogenicity of the species in Taiwan.

Analysis of Changes in Oil Palm Canopy Architecture From Basal Stem Rot Using Terrestrial Laser Scanner.

Basal stem rot (BSR), caused by the Ganoderma fungus, is an infectious disease that affects oil palm (Elaeis guineensis) plantations. BSR leads to a significant economic loss and reductions in yields of up to Malaysian Ringgit (RM) 1.5 billion (US$400 million) yearly. By 2020, the disease may affect ∼1.7 million tonnes of fresh fruit bunches. The plants appear symptomless in the early stages of infection, although most plants die after they are infected. Thus, early, accurate, and nondestructive disease detection is crucial to control the impact of the disease on yields. Terrestrial laser scanning (TLS) is an active remote-sensing, noncontact, cost-effective, precise, and user-friendly method. Through high-resolution scanning of a tree's dimension and morphology, TLS offers an accurate indicator for health and development. This study proposes an efficient image processing technique using point clouds obtained from TLS ground input data. A total of 40 samples (10 samples for each severity level) of oil palm trees were collected from 9-year-old trees using a ground-based laser scanner. Each tree was scanned four times at a distance of 1.5 m. The recorded laser scans were synched and merged to create a cluster of point clouds. An overhead two-dimensional image of the oil palm tree canopy was used to analyze three canopy architectures in terms of the number of pixels inside the crown (crown pixel), the degree of angle between fronds (frond angle), and the number of fronds (frond number). The results show that the crown pixel, frond angle, and frond number are significantly related and that the BSR severity levels are highly correlated (R2 = 0.76, P < 0.0001; R2 = 0.96, P < 0.0001; and R2 = 0.97, P < 0.0001, respectively). Analysis of variance followed post hoc tests by Student-Newman-Keuls (Newman-Keuls) and Dunnett for frond number presented the best results and showed that all levels were significantly different at a 5% significance level. Therefore, the earliest stage that a Ganoderma infection could be detected was mildly infected (T1). For frond angle, all post hoc tests showed consistent results, and all levels were significantly separated except for T0 and T1. By using the crown pixel parameter, healthy trees (T0) were separated from unhealthy trees (moderate infection [T2] and severe infection [T3]), although there was still some overlap with T1. Thus, Ganoderma infection could be detected as early as the T2 level by using the crown pixel and the frond angle parameters. It is hard to differentiate between T0 and T1, because during mild infection, the symptoms are highly similar. Meanwhile, T2 and T3 were placed in the same group, because they showed the same trend. This study demonstrates that the TLS is useful for detecting low-level infection as early as T1 (mild severity). TLS proved beneficial in managing oil palm plantation disease.

RNA-seq data of Ganoderma boninense at axenic culture condition and under in planta pathogen-oil palm (Elaeis guineensis Jacq.) interaction.

OBJECTIVE: Basal stem rot disease causes severe economic losses to oil palm production in South-east Asia and little is known on the pathogenicity of the pathogen, the basidiomyceteous Ganoderma boninense. Our data presented here aims to identify both the house-keeping and pathogenicity genes of G. boninense using Illumina sequencing reads. DESCRIPTION: The hemibiotroph G. boninense establishes via root contact during early stage of colonization and subsequently kills the host tissue as the disease progresses. Information on the pathogenicity factors/genes that causes BSR remain poorly understood. In addition, the molecular expressions corresponding to G. boninense growth and pathogenicity are not reported. Here, six transcriptome datasets of G. boninense from two contrasting conditions (three biological replicates per condition) are presented. The first datasets, collected from a 7-day-old axenic condition provide an insight onto genes responsible for sustenance, growth and development of G. boninense while datasets of the infecting G. boninense collected from oil palm-G. boninense pathosystem (in planta condition) at 1 month post-inoculation offer a comprehensive avenue to understand G. boninense pathogenesis and infection especially in regard to molecular mechanisms and pathways. Raw sequences deposited in Sequence Read Archive (SRA) are available at NCBI SRA portal with PRJNA514399, bioproject ID.

Simultaneous transcriptome analysis of oil palm clones and Phytophthora palmivora reveals oil palm defense strategies.

Phytophthora palmivora is an oomycete that causes oil palm bud rot disease. To understand the molecular mechanisms of this disease, palm clones with contrasting responses (Ortet 34, resistant and Ortet 57, susceptible) were inoculated with P. palmivora, and RNAseq gene expression analysis was performed. The transcriptome was obtained by sequencing using Illumina HiSeq2500 technology during the asymptomatic phase (24, 72 and 120 hours postinfection, hpi). A simultaneous analysis of differentially expressed gene (DEG) profiles in palm and P. palmivora was carried out. Additionally, Gene Ontology (GO) and gene network analysis revealed differences in the transcriptional profile of the two ortets, where a high specificity of the pathogen to colonize the susceptible ortet was found. The transcriptional analysis provided an overview of the genes involved in the recognition and signaling of this pathosystem, where different transcription factors, phytohormones, proteins associated with cell wall hardening and nitrogen metabolism contribute to the resistance of oil palm to P. palmivora. This research provides a description of the molecular response of oil palm to P. palmivora, thus becoming an important source of molecular markers for the study of genotypes resistant to bud rot disease.

Preparation of Chitosan-Hexaconazole Nanoparticles as Fungicide Nanodelivery System for Combating Ganoderma Disease in Oil Palm.

Fungicide is used to control fungal disease by destroying and inhibiting the fungus or fungal spores that cause the disease. However, failure to deliver fungicide to the disease region leads to ineffectiveness in the disease control. Hence, in the present study, nanotechnology has enabled the fungicide active agents (hexaconazole) to be encapsulated into chitosan nanoparticles with the aim of developing a fungicide nanodelivery system that can transport them more effectively to the target cells (Ganoderma fungus). A pathogenic fungus, Ganoderma boninense (G. boninense), is destructive to oil palm whereby it can cause significant loss to oil palm plantations located in the Southeast Asian countries, especially Malaysia and Indonesia. In regard to this matter, a series of chitosan nanoparticles loaded with the fungicide, hexaconazole, was prepared using various concentrations of crosslinking agent sodium tripolyphosphate (TPP). The resulting particle size revealed that the increase of the TPP concentration produced smaller particles. In addition, the in vitro fungicide released at pH 5.5 demonstrated that the fungicide from the nanoparticles was released in a sustainable manner with a prolonged release time up to 86 h. On another note, the in vitro antifungal studies established that smaller particle size leads to lower half maximum effective concentration (EC50) value, which indicates higher antifungal activity against G. boninense.

Elaboração e análise microbiológica de geleia de polpa de Mauritia flexuosa / Elaboration and microbiological analysis of Mauritia flexuosa polar jelly

A região dos cerrados é muito rica em espécies frutíferas nativas, oferecendo grande quantidade de frutas comestíveis, e dentre elas encontra-se o Buriti. Diante disso, objetivou-se com esta pesquisa elaborar e avaliar as características microbiológicas de geleia de polpa de buriti após 90 dias de estocagem a 25 ºC. Para a elaboração da geleia foi utilizada a proporção de 60 partes da mistura da polpa de buriti para 40 partes de açúcar. Foram realizadas as determinações de Coliformes totais e termotolerantes, Salmonella spp., mesófilos, bolores e leveduras após 1, 15, 30, 45, 60 e 90 dias de estocagem a 25 ºC. Os resultados indicaram que a geleia elaborada não apresentaram contaminação por bolores e leveduras, Salmonella spp., coliformes e mesófilos estando em consonância com os parâmetros exigidos pela legislação para geleias.

Microbial Diversity in Decaying Oil Palm Empty Fruit Bunches (OPEFB) and Isolation of Lignin-degrading Bacteria from a Tropical Environment.

Oil palm empty fruit bunches (OPEFB) are the most abundant, inexpensive, and environmentally friendly lignocellulosic biomass in Malaysia. Investigations on the microbial diversity of decaying OPEFB may reveal microbes with complex enzymes that have the potential to enhance the conversion of lignocellulose into second-generation biofuels as well as the production of other value-added products. In the present study, fungal and bacterial diversities in decaying OPEFB were identified using Illumina MiSeq sequencing of the V3 region of the 16S rRNA gene and V4 region of the 18S rRNA gene. Fungal diversity in decaying OPEFB was dominated by the phylum Ascomycota (14.43%), while most of the bacterial sequences retrieved belonged to Proteobacteria (76.71%). Three bacterial strains isolated from decaying OPEFB, designated as S18, S20, and S36, appeared to grow with extracted OPEFB-lignin and Kraft lignin (KL) as the sole carbon source. 16S rRNA gene sequencing identified the 3 isolates as Paenibacillus sp.. The molecular weight distribution of KL before and after degradation showed significant depolymerization when treated with bacterial strains S18, S20, and S36. The presence of low-molecular-weight lignin-related compounds, such as vanillin and 2-methoxyphenol derivatives, which were detected by a GC-MS analysis, confirmed the KL-degrading activities of isolated Paenibacillus strains.