Living jewels: iterative evolution of iridescent blue leaves from helicoidal cell walls.

BACKGROUND AND AIMS: Structural colour is responsible for the remarkable metallic blue colour seen in the leaves of several plants. Species belonging to only ten genera have been investigated to date, revealing four photonic structures responsible for structurally coloured leaves. One of these is the helicoidal cell wall, known to create structural colour in the leaf cells of five taxa. Here we investigate a broad selection of land plants to understand the phylogenetic distribution of this photonic structure in leaves. METHODS: We identified helicoidal structures in the leaf epidermal cells of 19 species using transmission electron microscopy. Pitch measurements of the helicoids were compared with the reflectance spectra of circularly polarized light from the cells to confirm the structure-colour relationship. RESULTS: By incorporating species examined with a polarizing filter, our results increase the number of taxa with photonic helicoidal cell walls to species belonging to at least 35 genera. These include 19 monocot genera, from the orders Asparagales (Orchidaceae) and Poales (Cyperaceae, Eriocaulaceae, Rapateaceae) and 16 fern genera, from the orders Marattiales (Marattiaceae), Schizaeales (Anemiaceae) and Polypodiales (Blechnaceae, Dryopteridaceae, Lomariopsidaceae, Polypodiaceae, Pteridaceae, Tectariaceae). CONCLUSIONS: Our investigation adds considerably to the recorded diversity of plants with structurally coloured leaves. The iterative evolution of photonic helicoidal walls has resulted in a broad phylogenetic distribution, centred on ferns and monocots. We speculate that the primary function of the helicoidal wall is to provide strength and support, so structural colour could have evolved as a potentially beneficial chance function of this structure.

The Role of Endophytes in Combating Fungal- and Bacterial-Induced Stress in Plants.

Plants are subjected to multifaceted stresses that significantly jeopardize crop production. Pathogenic microbes influence biotic stress in plants, which ultimately causes annual crop loss worldwide. Although the use of pesticides and fungicides can curb the proliferation of pathogens in plants and enhance crop production, they pollute the environment and cause several health issues in humans and animals. Hence, there is a need for alternative biocontrol agents that offer an eco-friendly mode of controlling plant diseases. This review discusses fungal- and bacterial-induced stress in plants, which causes various plant diseases, and the role of biocontrol defense mechanisms, for example, the production of hydrolytic enzymes, secondary metabolites, and siderophores by stress-tolerant fungi and bacteria to combat plant pathogens. It is observed that beneficial endophytes could sustain crop production and resolve the issues regarding crop yield caused by bacterial and fungal pathogens. The collated literature review indicates that future research is necessary to identify potential biocontrol agents that can minimize the utility of synthetic pesticides and increase the tenable agricultural production.

Identification of culturable petroleum-degrading bacteria and fungi from petroleum-contaminated sites in Brunei Darussalam.

Microbes that can be cultured and degrade petroleum are of particular interest for biotechnology such as bioremediation. This study aims to isolate and identify culturable petroleum-degrading bacteria and fungi from Brunei Darussalam, which has not previously been explored. A total of eight bacterial and nine fungal isolates that could degrade petroleum were obtained from petroleum-contaminated water or soil samples. DNA barcoding using 16S rRNA gene sequence identified five different bacterial genera which were Bacillus, Enterobacter, Micrococcus, Pseudoaltermonas and Pseudomonas. DNA barcoding using rRNA-ITS gene sequence identified nine different fungal taxa which were Aspergillus, Cladosporium, Exophiala, Flavodon, Hypocreales, Nectriaceae, Penicillium, Peniophora and Trichoderma. Biolog provided additional support to the identification of some isolates. This study is the first to report these unique microbes from Brunei Darussalam, which are of ecological and biotechnological value.

Controls on foliar nutrient and aluminium concentrations in a tropical tree flora: phylogeny, soil chemistry and interactions among elements.

Foliar elemental concentrations are predictors of life-history variation and contribute to spatial patterns in biogeochemical cycling. We examined the contributions of habitat association, local soil environment, and elemental interactions to variation in foliar elemental concentrations in tropical trees using methods that account for phylogeny. We sampled top-soils and leaves of 58 tropical trees in heath forest (HF) on nutrient-poor sand and mixed dipterocarp forest (MDF) on nutrient-rich clay soils. A phylogenetic generalized least squares method was used to determine how foliar nutrient and aluminium (Al) concentrations varied in response to habitat distribution, soil chemistry and other elemental concentrations. Foliar nitrogen (N) and Al concentrations were greater for specialists of MDF than for specialists of HF, while foliar calcium (Ca) concentrations showed the opposite trend. Foliar magnesium (Mg) concentrations were lower for generalists than for MDF specialists. Foliar element concentrations were correlated with fine-scale variation in soil chemistry in phylogenetically controlled analyses across species, but there was limited within-species plasticity in foliar elemental concentrations. Among Al accumulators, foliar Al concentration was positively associated with foliar Ca and Mg concentrations, and negatively associated with foliar phosphorus (P) concentrations. The Al-accumulation trait and relationships between foliar elemental and Al concentrations may contribute to species habitat partitioning and ecosystem-level differences in biogeochemical cycles.

Ecophysiological responses of native and introduced coastal tree species parasitized by Cassytha filiformis in Brunei.

Hemiparasitic Cassytha filiformis commonly infects native host (Dillenia suffruticosa and Melastoma malabathricum) and introduced host (Acacia auriculiformis and Acacia mangium) species in threatened heath forests in Brunei. This study aims to investigate the impact of parasitism on the ecophysiology of these host species. This study addresses the research gap in understanding the ecophysiology of C. filiformis-host associations, particularly when native and introduced hosts were infected. We generated CO2 and light response curves to examine the effects of increasing CO2 and light levels of infected and uninfected hosts and examined gaseous exchange, mineral nutrients, and secondary bioactive compounds of host-parasite associations. Infected hosts were negatively impacted by C. filiformis as exhibited in the CO2 and light response curves, with C. filiformis-native host association performing better than introduced species. There were no significant differences in photosynthetic parameters between infected and uninfected hosts, except in D. suffruticosa. Certain nutrient contents showed significant differences, but total N, Ca, and K in uninfected hosts were similar to infected hosts. Total phenols and tannins were significantly higher in introduced hosts than native hosts. Our findings asserted that this hemiparasitic vine relies on both its photosynthetic efficiency and nutrient acquisition from its hosts. The parasitism did not significantly hinder the ecophysiological performance of infected hosts, suggesting a plausible co-existence between the hosts and C. filiformis. This study provides essential ecophysiological information for future research on how C. filiformis can establish itself without negatively impacting the co-habitating native hosts.

Mangrove endophytic fungi: Biocontrol potential against Rhizoctonia solani and biofertilizers for fragrant rice cultivation.

The mangrove ecosystem has emerged as a fascinating source for exploring novel bioresources which have multiple applications in modern agriculture. This study evaluates the potential applications of mangrove endophytic fungi (MEF), such as biocontrol agents against Rhizoctonia solani and as biofertilizers for improving the yield of fragrant rice variety Malaysian Rice Quality 76 (MRQ76). Through the antagonism assays, it is observed that among the 14 MEF studied, 4 fungal isolates (Colletotrichum sp. MEFN02, Aspergillus sp. MEFN06, Annulohypoxylon sp. MEFX02 and Aspergillus sp. MEFX10) exhibited promising antagonistic effect against the pathogen R. solani compared to the chemical fungicide (Benomyl). These isolates also revealed significant production of enzymes, phytochemicals, indoleacetic acid (40.96 mg/mL) and ammonia (32.54 mg/mL) and displayed tolerance to salt and temperature stress up to 2000 mM and >40 °C respectively. Furthermore, employing the germination and pathogenicity test, inoculation of these endophytes showed lower percentage of disease severity index (DSI%) against R. solani, ranging from (24 %-46 %) in MRQ76 rice seedlings. The in-vivo experiments of soil and seed inoculation methods conducted under greenhouse conditions revealed that these endophytes enhanced plant growth (8-15 % increase) and increased crop yield (≥50 %) in comparison to control treatments. The current findings provide valuable insights into eco-friendly, cost-effective and sustainable alternatives for addressing R. solani infection and improving the agronomic performance of the fragrant rice cultivar MRQ76, contributing to food security.

Heavy metal tolerance and accumulation in the Brassica species (Brassica chinensis var. parachinensis and Brassica rapa L.): A pot experiment.

This study delves into the heavy metal tolerance and accumulation capabilities of Brassica chinensis var. parachinensis (B. chinensis) and Brassica rapa L. (B. rapa) in a pot experiment, specifically focusing on cadmium (Cd), chromium (Cr) and lead (Pb). Agricultural topsoils were spiked with varying concentrations of these heavy metals (0 mg/kg, 75 mg/kg, 150 mg/kg, 225 mg/kg and 300 mg/kg) for each element. The experiment involved cultivating 15 pots each of B. chinensis and B. rapa over 60 days. Results indicated that both Brassica species experienced delayed germination, with B. chinensis exhibiting a significant drop in germination percentage to 53 % at the highest concentration (300 mg/kg), while B. rapa showed a tendency for an increased germination percentage of up to 80 % at elevated metal concentrations; however, these differences were not statistically significant. Both B. chinensis and B. rapa demonstrated a stable decline in growth rate from 0.05 cm/day to 0.04 cm/day with increasing heavy metal concentrations, and the he reduction in relative growth rate was significant at the highest concentration compared to the control. The stress tolerance index revealed a significant decrease in plant heights for B. chinensis, in contrast to the stable performance of B. rapa, showcasing the tolerance of B. rapa to toxic conditions. Despite insignificant differences in fresh biomass due to metal treatments, B. chinensis consistently yielded higher biomass, yet it had a lower edible index due to its higher root biomass. Leaf areas increased significantly in both species at higher soil treatments, while root lengths remained unchanged, suggesting their resilience to elevated heavy metal concentrations. Analysis of plant tissues (leaves, stems and roots) using ICP-OES revealed that B. rapa accumulated the highest Cd concentration (864 mg/kg), whereas B. chinensis accumulated the highest Pb concentration (953 mg/kg) in root parts. Both species significantly accumulated Cr in roots, demonstrating a sequestration mechanism. These findings suggest that both species, particularly, B. rapa possess strong tolerance and accumulation capabilities for non-essential heavy metals, making them potential hyperaccumulators for green remediation techniques in toxic soil environments. Understanding the molecular mechanisms driving these responses and validating phytoremediation potential in real-world scenarios is essential for developing sustainable soil management practices.

Host Selectivity and Distribution of Cassytha filiformis in the Coastal Bornean Heath Forests.

We investigated the host range of Cassytha filiformis L. in the heath forests using six 50-metre transects. Sixteen shrubs and tree species were infected by C. filiformis vines, including two exotic Acacia species. This paper also examined the density and vigour of C. filiformis when infecting the two most preferred and common hosts, the heath native Dillenia suffruticosa (Griff. ex Hook. f. and Thomson) Martelli, and the invasive Acacia mangium Willd. The results suggested that C. filiformis has higher vigour when infecting native hosts than in exotic A. mangium albeit being not statistically significant. The long thread-like stems of parasite were present at relatively high density when infecting A. mangium, regardless of the host conditions. We also assessed the functionality of the haustoria on both D. suffruticosa and A. mangium using histological methods. It was found that C. filiformis can establish a true haustorial endophytic connection with studied hosts. Under controlled conditions, C. filiformis pose as a possible candidate for a biological control agent of A. mangium to curtail the fast spreading of this introduced species in tropical Borneo.

Comparative assessment of the heavy metal phytoextraction potential of vegetables from agricultural soils: A field experiment.

A field study was established to determine the phytoextraction potential of six vegetable species, namely Amaranthus viridis L., Basella alba L., Brassica chinensis var. Parachinensis, Brassica rapa L., Capsicum frutescens L., and Ocimum tenuiflorum L.. These edible plants were selected for their short growth cycles and high biomass production, which are some traits for efficient phytoremediation. Following acid digestion of the soil and vegetable samples using the USEPA 3050B acid digestion method, the extracts were analyzed for Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn using ICP-OES. Results in soil samples showed that the concentrations of both beneficial and essential heavy metals, and non-essential heavy metals are below the WHO, USEPA, and CCME soil quality guidelines. Al is one of the highest concentrations found in the soil samples but it tends to accumulate in the root part of all vegetable species compared to the aboveground parts. In general, B. rapa L. accumulated the highest level of Cd (0.4 mg/kg) and Pb (5.71 mg/kg), while B. alba L. accumulated the highest Cr (2.62 mg/kg) in all plant parts. The findings in this study indicated that Co, Cu, Fe, Mn and Zn were mostly accumulated in leaves of A. viridis L. (Co, Cu, Fe, Mn and Zn), B. alba L. (Co, Fe and Mn), B. chinensis (Mn and Zn) and O. tenuiflorum L. (Mn), and roots of C. frutescens L. (Co, Cu, Fe and Mn), B. alba L. (Co, Cu and Zn), A. viridis L. and B. chinensis (Cu and Fe) and B. rapa L. (Fe). Cr, Pb and Ni were significantly greater in B. alba L. (Cr) And B. rapa L. (Ni and Pb) roots. MTF >1 was observed in the roots of all species for Co, Cd, Zn, and Ni. BTC values varied between the different vegetable species with A. viridis L. having the greatest heavy metal mobility between its plant parts and the best heavy metal phytoextraction potential among other species. The PCA biplots showed that heavy metals were partitioned differently between various plant parts of the vegetable species and can be explained by the first two components (PC1 and PC2) which were associated with the root and/or leaf parts for most vegetable species.

Genetic structure and population history of a peat swamp forest tree species, Shorea albida (Dipterocarpaceae), in Brunei Darussalam.

Southeast Asia supports high biodiversity, in a mosaic of forest types formed by the expansion and contraction of habitats through past climate changes. Among the region's forest types, the geographical distribution of peat swamp forests has fluctuated intensely over the past 120,000 years. Most peat swamp forests in Southeast Asia are found in coastal regions and formed within the last 7,000 years after a decline in sea level. However, some peat swamps were initiated earlier on substrates of slightly higher elevation, and these peat swamps might have been refugia for peat swamp species in the last glacial period and the high sea level period. We assessed genetic diversity, genetic structure and divergence time of current genetic groups for Shorea albida in Brunei, an endemic tree species of Bornean peat swamp forests, using 18 microsatellite markers. Genetic diversity was not lower than has been found in other Shorea species, possibly because of the high density of S. albida in Brunei. Although overall genetic divergence between populations was low, two populations (Ingei and Labi Road 3) were distinct from the other populations. Analysis using DIYABC estimated that three genetic groups (Ingei, Labi Road 3 and others) diverged simultaneously from their ancestral population, whose effective size was very small, about 7,500 years ago, corresponding to a recent sea level peak in the Belait-Baram river basin. In that high sea level period, some higher-elevation lands remained, and peat formation had already started in this region. We propose that the current genetic structure of S. albida in Brunei was formed from small refugial populations that survived the period of higher sea level in these higher-elevation areas. Because of their relatively high genetic diversity, Brunei's S. albida populations should become an important genetic resource for the recovery of genetically healthy populations in other parts of northwest Borneo.

Evidence of foliar aluminium accumulation in local, regional and global datasets of wild plants.

• High foliar concentrations of aluminium (Al) have been reported in numerous plant species, but progress on the understanding of the functional significance of this trait is constrained by the absence of a quantitative analysis of its distribution among plant lineages and across biomes. • We constructed a global dataset of foliar Al and nutrient concentrations for 1044 plant species from literature sources and new data collections in Brunei Darussalam. • Our results provide statistical support for the existence of Al accumulators and non-Al accumulators in global, regional and local floras based on foliar Al concentrations. A value of 1 mg Al g(-1) leaf dry mass is a suitable threshold to distinguish between these two groups in a sample of species that lacks any geographical reference. However, a higher threshold foliar Al concentration is required to distinguish between Al accumulators in tropical (2.3-3.9 mg Al g(-1) leaf dry mass) than in temperate (1.1 mg Al g(-1) leaf dry mass) floras. There was a phylogenetic signal in the foliar concentrations of Al, but phylogeny did not explain the difference in the mean foliar Al concentration between tropical and temperate floras in a phylogenetically controlled analysis. • Phylogeny and soil chemistry are potential factors driving Al accumulation in certain groups of plants.

Genomic insights into rapid speciation within the world's largest tree genus Syzygium.

Species radiations, despite immense phenotypic variation, can be difficult to resolve phylogenetically when genetic change poorly matches the rapidity of diversification. Genomic potential furnished by palaeopolyploidy, and relative roles for adaptation, random drift and hybridisation in the apportionment of genetic variation, remain poorly understood factors. Here, we study these aspects in a model radiation, Syzygium, the most species-rich tree genus worldwide. Genomes of 182 distinct species and 58 unidentified taxa are compared against a chromosome-level reference genome of the sea apple, Syzygium grande. We show that while Syzygium shares an ancient genome doubling event with other Myrtales, little evidence exists for recent polyploidy events. Phylogenomics confirms that Syzygium originated in Australia-New Guinea and diversified in multiple migrations, eastward to the Pacific and westward to India and Africa, in bursts of speciation visible as poorly resolved branches on phylogenies. Furthermore, some sublineages demonstrate genomic clines that recapitulate cladogenetic events, suggesting that stepwise geographic speciation, a neutral process, has been important in Syzygium diversification.

Transfer of heavy metals from soils to curly mustard (Brassica juncea (L.) Czern.) grown in an agricultural farm in Brunei Darussalam.

Determination of heavy metal concentrations in vegetables and agricultural soils is crucial because high levels of heavy metals could affect soil quality, crop production and safe consumption of crops. A field study was conducted to determine the heavy metal concentrations and their transfer from agricultural soils to different parts (leaf, stem, and root) of Brassica juncea (L.) Czern. In addition, potential health risks of contamination in the vegetables grown in the field were evaluated. Acid digestion method USEPA 3050B in combination with ICP-OES were used to analyze heavy metal (Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn) contents in both pre- and post-harvest soils and vegetable samples. Results showed that none of the heavy metals in soils had concentrations above the maximum safety limits based on the WHO, USEPA and CCME guidelines. Calculated metal transfer factor (MTF >1) showed B. juncea accumulated Cd, Co, Ni, Pb and Zn in leaves, stems and roots, but Cu and Mn, as well as Cr were only accumulated in stems and roots, respectively. There were variations in heavy metal contents between the different parts of B. juncea, but only Cd and Pb contents were above the maximum allowable limit recommended by FAO/WHO. PCA analysis was able to identify 4 major components corresponding to 38.38%, 28.98%, 14.39% and 10.67% of the total variance and PC1 was clearly associated to leaves of B. juncea. Based on the MTF values, only Cd was found to have a value of HRI >1 compared to the other heavy metals, implying potential health risk associated with long-term ingestion of the vegetable.

Vegetative Propagation of Hoya imperialis and Hoya coronaria by Stem Cutting and Micropropagation.

Hoya imperialis (H. imperialis) and H. coronaria (Apocynaceae) are known to have ornamental value due to their beautiful flowers; however, the feasibility of propagating these plants have not been reported despite the wild populations in Brunei Darussalam being highly threatened due to habitat loss and overcollection. Thus, the present study aimed to conduct a preliminary study of the feasibility of two alternative propagation methods, stem cutting and micropropagation, as a potential approach for their ex situ conservation. Hoya stem cuttings were treated with either indole-3-butyric acid (IBA) or 1-naphthaleneacetic acid (NAA) (0-2000 mg/L), and then propagated onto a mixture of peat moss and perlite. For micropropagation, Hoya leaf explants were cultured onto Murashige and Skoog (MS) agar media that were supplemented with IBA and/or kinetin (KN) (0-10.0 mg/L). This present study shows that both Hoya species were successfully propagated by stem cutting even without hormone treatment. However, interestingly, in H. imperialis, when compared with control, the mean number of new leaves (6.3 ± 1.0) and the mean relative growth rate (RGR) based on stem diameter (0.004 ± 0.0007 cm cm-1 day-1) significantly increased when treated with 500 mg/L NAA and 2000 mg/L IBA, respectively. Meanwhile, in H. coronaria, significantly higher mean number of roots was achieved by treating with 1000 mg/L NAA (16.6 ± 1.4) or 2000 mg/L IBA (17.5 ± 2.7) compared with control. For micropropagation, callus induction was not promising and could only be observed at specific concentrations of both IBA and KN, with H. imperialis appearing to be more responsive towards these hormones in comparison to H. coronaria. The present study showed that stem cutting appeared more feasible in propagating both Hoya species.

Sex is determined by XY chromosomes across the radiation of dioecious Nepenthes pitcher plants.

Species with separate sexes (dioecy) are a minority among flowering plants, but dioecy has evolved multiple times independently in their history. The sex-determination system and sex-linked genomic regions are currently identified in a limited number of dioecious plants only. Here, we study the sex-determination system in a genus of dioecious plants that lack heteromorphic sex chromosomes and are not amenable to controlled breeding: Nepenthes pitcher plants. We genotyped wild populations of flowering males and females of three Nepenthes taxa using ddRAD-seq and sequenced a male inflorescence transcriptome. We developed a statistical tool (privacy rarefaction) to distinguish true sex specificity from stochastic noise in read coverage of sequencing data from wild populations and identified male-specific loci and XY-patterned single nucleotide polymorphsims (SNPs) in all three Nepenthes taxa, suggesting the presence of homomorphic XY sex chromosomes. The male-specific region of the Y chromosome showed little conservation among the three taxa, except for the essential pollen development gene DYT1 that was confirmed as male specific by PCR in additional Nepenthes taxa. Hence, dioecy and part of the male-specific region of the Nepenthes Y-chromosomes likely have a single evolutionary origin.

Field methods for sampling tree height for tropical forest biomass estimation.

Quantifying the relationship between tree diameter and height is a key component of efforts to estimate biomass and carbon stocks in tropical forests. Although substantial site-to-site variation in height-diameter allometries has been documented, the time consuming nature of measuring all tree heights in an inventory plot means that most studies do not include height, or else use generic pan-tropical or regional allometric equations to estimate height.Using a pan-tropical dataset of 73 plots where at least 150 trees had in-field ground-based height measurements, we examined how the number of trees sampled affects the performance of locally derived height-diameter allometries, and evaluated the performance of different methods for sampling trees for height measurement.Using cross-validation, we found that allometries constructed with just 20 locally measured values could often predict tree height with lower error than regional or climate-based allometries (mean reduction in prediction error = 0.46 m). The predictive performance of locally derived allometries improved with sample size, but with diminishing returns in performance gains when more than 40 trees were sampled. Estimates of stand-level biomass produced using local allometries to estimate tree height show no over- or under-estimation bias when compared with biomass estimates using field measured heights. We evaluated five strategies to sample trees for height measurement, and found that sampling strategies that included measuring the heights of the ten largest diameter trees in a plot outperformed (in terms of resulting in local height-diameter models with low height prediction error) entirely random or diameter size-class stratified approaches.Our results indicate that even limited sampling of heights can be used to refine height-diameter allometries. We recommend aiming for a conservative threshold of sampling 50 trees per location for height measurement, and including the ten trees with the largest diameter in this sample.

Author Correction: Long-term carbon sink in Borneo's forests halted by drought and vulnerable to edges.

The original version of this Article contained an error in the third sentence of the abstract and incorrectly read "Here, using long-term plot monitoring records of up to half a century, we find that intact forests in Borneo gained 0.43 Mg C ha-1 year-1 (95% CI 0.14-0.72, mean period 1988-2010) above-ground live biomass", rather than the correct "Here, using long-term plot monitoring records of up to half a century, we find that intact forests in Borneo gained 0.43 Mg C ha-1 year-1 (95% CI 0.14-0.72, mean period 1988-2010) in above-ground live biomass carbon". This has now been corrected in both the PDF and HTML versions of the Article.

Long-term carbon sink in Borneo's forests halted by drought and vulnerable to edge effects.

Less than half of anthropogenic carbon dioxide emissions remain in the atmosphere. While carbon balance models imply large carbon uptake in tropical forests, direct on-the-ground observations are still lacking in Southeast Asia. Here, using long-term plot monitoring records of up to half a century, we find that intact forests in Borneo gained 0.43 Mg C ha-1 per year (95% CI 0.14-0.72, mean period 1988-2010) above-ground live biomass. These results closely match those from African and Amazonian plot networks, suggesting that the world's remaining intact tropical forests are now en masse out-of-equilibrium. Although both pan-tropical and long-term, the sink in remaining intact forests appears vulnerable to climate and land use changes. Across Borneo the 1997-1998 El Niño drought temporarily halted the carbon sink by increasing tree mortality, while fragmentation persistently offset the sink and turned many edge-affected forests into a carbon source to the atmosphere.