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  1. Kobayashi MJ, Ng KKS, Lee SL, Muhammad N, Tani N
    Am J Bot, 2020 11;107(11):1491-1503.
    PMID: 33190268 DOI: 10.1002/ajb2.1557
    PREMISE: Leaf phenology is an essential developmental process in trees and an important component in understanding climate change. However, little is known about the regulation of leaf phenology in tropical trees.

    METHODS: To understand the regulation by temperature of leaf phenology in tropical trees, we performed daily observations of leaf production under rainfall-independent conditions using saplings of Shorea leprosula and Neobalanocarpus heimii, both species of Dipterocarpaceae, a dominant tree family of Southeast Asia. We analyzed the time-series data obtained using empirical dynamic modeling (EDM) and conducted growth chamber experiments.

    RESULTS: Leaf production by dipterocarps fluctuated in the absence of fluctuation in rainfall, and the peaks of leaf production were more frequent than those of day length, suggesting that leaf production cannot be fully explained by these environmental factors, although they have been proposed as regulators of leaf phenology in dipterocarps. Instead, EDM suggested a causal relationship between temperature and leaf production in dipterocarps. Leaf production by N. heimii saplings in chambers significantly increased when temperature was increased after long-term low-temperature treatment. This increase in leaf production was observed even when only nighttime temperature was elevated, suggesting that the effect of temperature on development is not mediated by photosynthesis.

    CONCLUSIONS: Because seasonal variation in temperature in the tropics is small, effects on leaf phenology have been overlooked. However, our results suggest that temperature is a regulator of leaf phenology in dipterocarps. This information will contribute to better understanding of the effects of climate change in the tropics.

  2. Takeuchi Y, Chaffron S, Salcher MM, Shimizu-Inatsugi R, Kobayashi MJ, Diway B, et al.
    Syst Appl Microbiol, 2015 Jul;38(5):330-9.
    PMID: 26138047 DOI: 10.1016/j.syapm.2015.05.006
    Pitchers are modified leaves used by carnivorous plants for trapping prey. Their fluids contain digestive enzymes from the plant and they harbor abundant microbes. In this study, the diversity of bacterial communities was assessed in Nepenthes pitcher fluids and the composition of the bacterial community was compared to that in other environments, including the phyllosphere of Arabidopsis, animal guts and another pitcher plant, Sarracenia. Diversity was measured by 454 pyrosequencing of 16S rRNA gene amplicons. A total of 232,823 sequences were obtained after chimera and singleton removal that clustered into 3260 distinct operational taxonomic units (OTUs) (3% dissimilarity), which were taxonomically distributed over 17 phyla, 25 classes, 45 orders, 100 families, and 195 genera. Pyrosequencing and fluorescence in situ hybridization yielded similar estimates of community composition. Most pitchers contained high proportions of unique OTUs, and only 22 OTUs (<0.6%) were shared by ≥14/16 samples, suggesting a unique bacterial assemblage in each pitcher at the OTU level. Diversity analysis at the class level revealed that the bacterial communities of both opened and unopened pitchers were most similar to that of Sarracenia and to that in the phyllosphere. Therefore, the bacterial community in pitchers may be formed by environmental filtering and/or by phyllosphere bacteria.
  3. Suhaimi AH, Kobayashi MJ, Satake A, Ng CC, Lee SL, Muhammad N, et al.
    PeerJ, 2023;11:e16368.
    PMID: 38047035 DOI: 10.7717/peerj.16368
    Climatic factors have commonly been attributed as the trigger of general flowering, a unique community-level mass flowering phenomenon involving most dipterocarp species that forms the foundation of Southeast Asian tropical rainforests. This intriguing flowering event is often succeeded by mast fruiting, which provides a temporary yet substantial burst of food resources for animals, particularly frugivores. However, the physiological mechanism that triggers general flowering, particularly in dipterocarp species, is not well understood largely due to its irregular and unpredictable occurrences in the tall and dense forests. To shed light on this mechanism, we employed ecological transcriptomic analyses on an RNA-seq dataset of a general flowering species, Shorea curtisii (Dipterocarpaceae), sequenced from leaves and buds collected at multiple vegetative and flowering phenological stages. We assembled 64,219 unigenes from the transcriptome of which 1,730 and 3,559 were differentially expressed in the leaf and the bud, respectively. Differentially expressed unigene clusters were found to be enriched with homologs of Arabidopsis thaliana genes associated with response to biotic and abiotic stresses, nutrient level, and hormonal treatments. When combined with rainfall data, our transcriptome data reveals that the trees were responding to a brief period of drought prior to the elevated expression of key floral promoters and followed by differential expression of unigenes that indicates physiological changes associated with the transition from vegetative to reproductive stages. Our study is timely for a representative general flowering dipterocarp species that occurs in forests that are under the constant threat of deforestation and climate change as it pinpoints important climate sensitive and flowering-related homologs and offers a glimpse into the cascade of gene expression before and after the onset of floral initiation.
  4. Ng KKS, Kobayashi MJ, Fawcett JA, Hatakeyama M, Paape T, Ng CH, et al.
    Commun Biol, 2021 Oct 07;4(1):1166.
    PMID: 34620991 DOI: 10.1038/s42003-021-02682-1
    Hyperdiverse tropical rainforests, such as the aseasonal forests in Southeast Asia, are supported by high annual rainfall. Its canopy is dominated by the species-rich tree family of Dipterocarpaceae (Asian dipterocarps), which has both ecological (e.g., supports flora and fauna) and economical (e.g., timber production) importance. Recent ecological studies suggested that rare irregular drought events may be an environmental stress and signal for the tropical trees. We assembled the genome of a widespread but near threatened dipterocarp, Shorea leprosula, and analyzed the transcriptome sequences of ten dipterocarp species representing seven genera. Comparative genomic and molecular dating analyses suggested a whole-genome duplication close to the Cretaceous-Paleogene extinction event followed by the diversification of major dipterocarp lineages (i.e. Dipterocarpoideae). Interestingly, the retained duplicated genes were enriched for genes upregulated by no-irrigation treatment. These findings provide molecular support for the relevance of drought for tropical trees despite the lack of an annual dry season.
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