Yeasts from phylloplane and their capability to produce indole-3-acetic acid

Yeasts were isolated from the phylloplane of various plant species collected from seven provinces in Thailand. A total of 114 yeast strains and 10 strains of a yeast-like fungus were obtained by enrichment isolation from 91 out of 97 leaf samples (93.8?%). On the basis of the D1/D2 domain of the large subunit rRNA gene sequence similarity, 98 strains were identified to be of 36 yeast species in 18 genera belonging to Ascomycota viz. Candida, Clavispora, Cyberlindnera, Debaryomyces, Hanseniaspora, Hyphopichia, Kazachstania, Kluyveromyces, Kodamaea, Lachancea, Metschnikowia, Meyrozyma, Pichia, Starmerella, Torulaspora and Wickerhamomyces, and to Basidiomycota viz. Sporidiobolus and Trichosporon. Three strains were found to represent two novels Candida species which were previously described as C. sirachaensis and C. sakaeoensis. Ten strains of yeast-like fungus were identified as Aureobasidium pullulans of the phylum Ascomycota. Ascomycetous yeast species accounted altogether for 98.0?% of the 98 strains. The prevalent species was Candida tropicalis with a low frequency of isolation (14.3?%). Diversity of yeasts other than ballistoconidium-forming yeast in phylloplane in a tropical country in Asia has been reported for the first time. All strains obtained were accessed for the capability to produce IAA and result revealed that 39 strains in 20 species, one strain each of an undescribed and a novel species, and two unidentified strains showed the capability of producing IAA when cultivated in yeast extract peptone dextrose broth supplemented with 0.1?% l-tryptophan. All five strains of Candida maltosa produced relatively high concentrations of IAA.     

Diversity of culturable yeasts in phylloplane of sugarcane in Thailand and their capability to produce indole-3-acetic acid

Yeasts were isolated by the enrichment technique from the phylloplane of 94 samples of sugarcane leaf collected from seven provinces in Thailand. All sugarcane leaf samples contained yeasts and 158 yeast strains were obtained. On the basis of the D1/D2 domain of the large subunit rRNA gene sequence analysis, 144 strains were identified to 24 known species in 14 genera belonging to the Ascomycota viz. Candida akabanensis, Candida dendronema, Candida mesorugosa, Candida michaelii, Candida nivariensis, Candida rugosa, Candida orthopsilosis, Candida quercitrusa, Candida tropicalis, Candida xylopsoci, Cyberlindnera fabianii, Cyberlindnera rhodanensis, Debaryomyces nepalensis, Hannaella aff. coprosmaensis, Hanseniaspora guilliermondii, Kluyveromyces marxianus, Lachancea thermotolerans, Lodderomyces elongisporus, Metschnikowia koreensis, Meyerozyma caribbica, Millerozyma koratensis, Pichia kudriavzevii, Torulaspora delbrueckii and Wickerhamomyces edaphicus, and 12 species in six genera of the Basidiomycota viz . Cryptococcus flavescens, Cryptococcus laurentii, Cryptococcus rajasthanensis, Kwoniella heveanensis, Rhodosporidium fluviale, Rhodosporidium paludigenum, Rhodotorula mucilaginosa, Rhodotorula sesimbrana, Rhodotorula taiwanensis, Sporidiobolus ruineniae, Sporobolomyces carnicolor and Sporobolomyces nylandii. Seven strains were identical or similar to four undescribed species. Another seven strains represented four novels species in the genus Metschnikowia, Nakazawaea, Wickerhamomyces and Yamadazyma. The results revealed 69 % of the isolated strains were ascomycete yeasts and 31 % were basidiomycete yeast. The most prevalent species was M. caribbica with a 23 % frequency of occurrence followed by Rh. taiwanensis (11 %) and C. tropicalis (10 %). All strains were assessed for indole-3-acetic acid (IAA) producing capability showing that 69 strains had the capability of producing IAA when cultivated in yeast extract peptone dextrose broth supplemented with 1 g/L l-tryptophan. The highest IAA concentration of 565.1 mg/L was produced by R. fluviale DMKU-RK253.     

Carbon fluxes from a tropical peat swamp forest floor

A tropical ombrotrophic peatland ecosystem is one of the largest terrestrial carbon stores. Flux rates of carbon dioxide (CO₂) and methane (CH₄) were studied at various peat water table depths in a mixed‐type peat swamp forest floor in Central Kalimantan, Indonesia. Temporary gas fluxes on microtopographically differing hummock and hollow peat surfaces were combined with peat water table data to produce annual cumulative flux estimates. Hummocks formed mainly from living and dead tree roots and decaying debris maintained a relatively steady CO₂ emission rate regardless of the water table position in peat. In nearly vegetation‐free hollows, CO₂ emission rates were progressively smaller as the water table rose towards the peat surface. Methane emissions from the peat surface remained small and were detected only in water‐saturated peat. By applying long‐term peat water table data, annual gas emissions from the peat swamp forest floor were estimated to be 3493±316 g CO₂ m^?2 and less than 1.36±0.57 g CH₄ m^?2. On the basis of the carbon emitted, CO₂ is clearly a more important greenhouse gas than CH₄. CO₂ emissions from peat are the highest during the dry season, when the oxic peat layer is at its thickest because of water table lowering.     

Surface peat structure and chemistry in a tropical peat swamp forest

Background and Aims

Tropical peat swamp forests (PSF) are great stores of terrestrial carbon and host unique biodiversity. Despite their importance for carbon accounting, the peat characteristics are sparsely studied, and the effect of microtopography on peat properties has not been reported before.

Methods

We compared PSF peat soil characteristics down to 70 cm under differing microtopographical conditions and hydrology.

Results

Long-term water table level data combined with the data from peat structure and chemistry analyses showed differences in most of the measured properties between hummocks and hollows. Decomposition degree was lowest at hummock and hollow surfaces while bulk density and C content increased towards deeper peat. Ash, P, K, Ca and Mg had highest concentrations on hummock surfaces with declining trend downwards, whereas N had no clear concentration pattern along the elevation gradient.

Conclusions

The microtopographical features may not only differ in regards to the water table-induced oxygen conditions but also due to differences in nutrient dynamics.     

Interdependence of peat and vegetation in a tropical peat swamp forest

The visual uniformity of tropical peat swamp forest masks the considerable variation in forest structure that has evolved in response to differences and changes in peat characteristics over many millennia. Details are presented of forest structure and tree composition of the principal peat swamp forest types in the upper catchment of Sungai Sebangau, Central Kalimantan, Indonesia, in relation to thickness and hydrology of the peat. Consideration is given to data on peat geochemistry and age of peat that provide evidence of the ombrotrophic nature of this vast peatland and its mode of formation. The future sustainability of this ecosystem is predicted from information available on climate change and human impact in this region.     

Identification of enzymes involved in indole-3-acetic acid degradation

Strains of Bradyrhizobium japonicum with the ability to catabolize indole-3-acetic acid (IAA) and strains of B. japonicum, Rhizobium loti, and Rhizobium galegae, unable to catabolize IAA, were analyzed for enzymes involved in the pathway for IAA degradation. Two enzymes having isatin as substrate were detected. An isatin amidohydrolase catalyzing the hydrolysis of isatin into isatinic acid was found in some B. japonicum strains and in two Rhizobium species, R loti and R. galegae. The enzyme was inducible (4–5-fold) by its substrate, isatin, and the partially purified enzyme from R. loti showed an apparent K_M of 11 M for isatin. A NADPH-dependent isatin reductase was measured in extracts from a strain of B. japonicum lacking the isatin amidohydrolase. The structure of the reaction product, dioxindole was verified by NMR spectroscopy. Isatin reductase activity was also detected in extracts of dry pea seeds, and present in at least two isoforms. A low K_M of 10 M for isatin was found with a partially purified preparation of the pea enzyme. The presence of such an enzyme activity in pea indicates dioxindole and isatin as possible intermediates in IAA degradation in pea.     

Carbon dioxide balance of a tropical peat swamp forest in Kalimantan, Indonesia

Tropical peatlands, which coexist with swamp forests, have accumulated vast amounts of carbon as soil organic matter. Since the 1970s, however, deforestation and drainage have progressed on an enormous scale. In addition, El Niño and Southern Oscillation (ENSO) drought and large‐scale fires, which grow larger under the drought condition, are accelerating peatland devastation. That devastation enhances decomposition of soil organic matter and increases the carbon release to the atmosphere as CO₂. This phenomenon suggests that tropical peatlands have already become a large CO₂ source, but related quantitative information is limited. Therefore, we evaluated the CO₂ balance of a tropical peat swamp forest in Central Kalimantan, Indonesia, using 3 years of CO₂ fluxes measured using the eddy covariance technique from 2002 through 2004. The forest was disturbed by drainage; consequently, groundwater level (GL) was reduced. The net ecosystem CO₂ production (NEP) measurements showed seasonal variation, which was slightly positive or almost zero in the early dry season, and most‐negative late in the dry season or early the rainy season. This seasonality is attributable to the seasonal pattern of climate, tree phenology and fires. Slightly positive NEP resulted from smaller ecosystem respiration (RE) and larger gross primary production (GPP) under conditions of high photosynthetic photon flux density (PPFD) and large leaf area index (LAI). The most‐negative NEP resulted from smaller GPP and larger RE. The smaller GPP was related to high vapor pressure deficit (VPD), small LAI and low PPFD because of smoke from fires. The larger RE was related to low GL. Annual NEP values were estimated respectively as −602, −382 and −313 g C m⁻² yr⁻¹ for 2002, 2003 and 2004. These negative NEP values show that the tropical peat swamp forest, disturbed by drainage, functioned as a CO₂ source. That source intensity was highest in 2002, an ENSO year, mainly because of low PPFD caused by dense smoke emitted from large fires.     

Orchid-associated bacteria produce indole-3-acetic acid,promote seed germination,and increase their microbial yield in response to exogenous auxin

Germination of orchid seeds is a complex process. In this paper we focus on interactions between the host-plant and its bacterial partners via indole-3-acetic acid (IAA). Originally isolated from the roots of the epiphytic orchid Dendrobium moschatum, the strains of Rhizobium, Microbacterium, Sphingomonas, and Mycobacterium genera were among the most active IAA producers. Addition of exogenous tryptophan significantly enhanced auxin formation both in mineral and complex media. The presence of IAA and indole-3-acetaldehyde was confirmed by HPLC. Indole-3-pyruvic and indole-3-lactic acids were also detected in supernatants of culture filtrates of Sphingomonas sp., Rhizobium sp., and Microbacterium sp., while indole-3-acetamide was identified only in Mycobacterium sp. Some concentration- and strain-dependent effects of exogenous IAA on bacterial development were also established. Treatment of the cultures with 10 and 100 μg/ml of auxin resulted in an increase in microbial yield. None of the investigated strains was able to utilize IAA as a source of carbon and energy. Furthermore, inoculation of D. moschatum seeds with Sphingomonas sp. and Mycobacterium sp. resulted in considerable enhancement of orchid seeds germination. This growth-promoting activity was observed in the absence of any plant growth stimulators or mycorrhizal fungi, usually required for orchid germination.     

Leaf litter decomposition in a tropical peat swamp forest in Peninsular Malaysia

It has long been assumed that the peat underlying tropical peat swamp forests accumulates because the extreme conditions (water logged, nutrient poor, anaerobic and acidic—pH 2.9–3.5) impede microbial activity. Litterbag studies in a tropical Malaysian peat swamp (North Selangor peat swamp forest) showed that although the sclerophyllous, toxic leaves of endemic peat forest plants (Macaranga pruinosa, Campnosperma coriaceum, Pandanus atrocarpus, Stenochlaena palustris) were barely decomposed by bacteria and fungi (decay rates of only 0.0006–0.0016 k day⁻¹), leaves of M. tanarius, a secondary forest species were almost completely decomposed (decay rates of 0.0047–0.005 k day⁻¹) after 1 year. Thus it is intrinsic properties of the leaves (that are adaptations to deter herbivory in the nutrient poor environment) that impede microbial breakdown. The water of the peat swamp was very high in dissolved organic carbon (70–84 mg l⁻¹ DOC). Laboratory studies revealed initial rapid leaching of DOC from leaves (up to 1,720 mg l⁻¹ from 4 g of leaves in 7 days), but the DOC levels then fell rapidly. The leaching of DOC resulted in weight loss but the physical structure of the leaves remained intact. It is suggested that the DOC is used as a substrate for microbial growth hence lowering the concentration of DOC in the water and transferring energy from the leaves to other trophic levels. This would explain how nutrient poor tropical peatswamps support diverse, abundant flora and fauna despite low nutrient levels and lack of rapid litter cycling such as occurs in other types of tropical rainforests.     

Microtopographic specialization and flexibility in tropical peat swamp forest tree species

Tropical tree species distributions are determined by a wide range of biotic and abiotic factors, including topography and hydrology. Tropical peat swamp forests (TPSFs) are characterized in part by small‐scale variations in topography (‘hummocks’ and ‘hollows’) that create distinct microhabitats and thus may contribute to niche diversification among TPSF tree species. Using tree elevations calibrated to daily peat water levels collected using a data logger and a permutation test, we evaluated topographical microhabitat preferences for 21 tree species in a relatively undisturbed TPSF in Central Kalimantan, Indonesia, to determine whether these species show preferential association with hummocks or hollows and to quantify the prevalence of microhabitat specialization among them. Only one species, Tetractomia tetrandrum, emerged as a hollow specialist, with no hummock specialists among the species tested. The remaining 20 species, including Psydrax dicoccos, which had the lowest mean observed elevation, and Maasia hypoleuca, which had the highest mean observed elevation, showed no clear microtopographic preference. This suggests that many TPSF species may be resilient to the natural hydrologic variations that occur in relatively intact peat swamp forests. Such studies of microtopographic preferences of tree species in TPSF and other wetland forest ecosystems can inform selection of tree species for reforestation projects, and potentially also provide information on how future climate change may impact these habitats and their resident tree species.     

Biogeochemical properties of a tropical swamp forest ecosystem in southern Thailand

The distributions of organic matter in the tropical swamps in southern Thailand are reported. The concentrations of particulate and dissolved organic carbon (POC and DOC) in the Bang Nara River, which drains swamp forests and nearby paddy fields, were 2.9 ± 2.0 and 6.2 ± 1.3 mg C l⁻¹, respectively. Although the variation was large, DOC concentration in the Bang Nara River seemed to be higher than POC in November 1992 (DOC/POC ratio, 2.8 ± 2.2). River waters from the upland areas were characterized by low POC and DOC concentrations as compared with Bang Nara River water. The δ¹³C values of POC and river sediments were useful to distinguish between organic matter originating in upland and swamp areas. It is suggested that the distributions of organic matter and its isotopic composition reflect the difference in drainage characteristics between lowland swamp and upland areas. Isotopic analyses of plant leaves and soils revealed that the swamp forest ecosystems were characterized by low δ¹³C and low δ¹⁵N values, which suggested low efficiency of water use by plants and large contributions of atmospheric deposition of nitrogen, respectively. Although CO₂ recycling in the forest might be an important factor determining the δ¹³C values of understory plants, the main process in carbon metabolism of tropical swamp forests would be CO₂ exchange between the atmosphere and forest canopy. Received: May 1, 2001 / Accepted: September 28, 2001     

Identification of oxindole-3-acetic acid,and metabolic conversion of indole-3-acetic acid to oxindole-3-acetic acid in Pinus sylvestris seeds

Oxindole-3-acetic acid (OxIAA) has been identified in germinating seeds of Scots pine (Pinus sylvestris) using gas chromatography-mass spectrometry. Seeds germinated for 5 d contained 2.7 ng OxIAA·g^-1 (dry weight) whereas ungerminated seeds contained 0.2 ng·g^-1. Isotopically labelled OxIAA was formed in seeds incubated with [1-¹⁴C]-, [2-¹⁴C]- or [²H₅]indole-3-acetic acid.Abbreviations DDC sodium diethyldithiocarbamate - GC gas chromatography - HPLC high-performance liquid chromatography - IAA indole-3-acetic acid - MS mass spectrometry - OxIAA oxindole-3-acetic acid - PVP polyvinylpyrrolidone - TMS trimethylsilyl     

Organic matter dynamics control plant species coexistence in a tropical peat swamp forest

We studied the relationship between the coexistence of tree species and the dynamics of organic matter in forests. A tropical peat swamp forest was selected as a model ecosystem, where abiotic factors, such as geological topography or parent rock types, are homogeneous and only biological processes create habitat heterogeneity. The temporal or spatial variation of the ground elevation of peat soils is mainly caused by changes in the balance between organic matter inputs to soils and decomposition, which is affected by the growth and death of influential trees. To clarify the processes of elevation dynamics, we measured the microtopography around some tree groups, estimated organic matter (in the form of litter and roots) in soils under three kinds of microtopographic conditions, measured decomposition rates and detected dominant species' shifting distribution patterns in different stages of growth in relation to the locations of tree groups creating specific microtopographic conditions. We found that growth or death of buttressed trees has the greatest effects on the rising or sinking of ground surfaces through changes in litter supply and root production. We discuss here the possibility of extending our model to other forest types.     

Changes in indole-3-acetic acid,indole-3-acetic acid oxidase,and peroxidase isoenzymes in the seeds of developing peach fruits

Changes in indole-3-acetic acid (IAA) content of peach (Prunus persica L. Batsch cv. Merry) seeds were followed during fruit development. The highest concentration of IAA, 2.7 g/g fresh weight, was found at the beginning of Stage III of fruit development, approximately 50–60 days after anthesis. The IAA-decarboxylating capacity of crude extracts of seeds was also greatest at 55–60 days after anthesis. Four soluble peroxidase isoenzymes were found on anionic electrophoresis. There were no marked changes in two isoenzymes (R _f 0.23 and 0.51), which were present in all three stages of fruit growth. There was a marked increase in a band atR _f 0.59 between Stages II and III, and a decrease in a band atR _f 0.68 from Stages II to III. Neither band (R _f 0.59 and 0.68) was present at Stage I.     

Transport of the two natural auxins, indole-3-butyric acid and indole-3-acetic acid, in Arabidopsis

Polar transport of the natural auxin indole-3-acetic acid (IAA) is important in a number of plant developmental processes. However, few studies have investigated the polar transport of other endogenous auxins, such as indole-3-butyric acid (IBA), in Arabidopsis. This study details the similarities and differences between IBA and IAA transport in several tissues of Arabidopsis. In the inflorescence axis, no significant IBA movement was detected, whereas IAA is transported in a basipetal direction from the meristem tip. In young seedlings, both IBA and IAA were transported only in a basipetal direction in the hypocotyl. In roots, both auxins moved in two distinct polarities and in specific tissues. The kinetics of IBA and IAA transport appear similar, with transport rates of 8 to 10 mm per hour. In addition, IBA transport, like IAA transport, is saturable at high concentrations of auxin, suggesting that IBA transport is protein mediated. Interestingly, IAA efflux inhibitors and mutations in genes encoding putative IAA transport proteins reduce IAA transport but do not alter IBA movement, suggesting that different auxin transport protein complexes are likely to mediate IBA and IAA transport. Finally, the physiological effects of IBA and IAA on hypocotyl elongation under several light conditions were examined and analyzed in the context of the differences in IBA and IAA transport. Together, these results present a detailed picture of IBA transport and provide the basis for a better understanding of the transport of these two endogenous auxins.     

Metabolism of indole-3-acetic acid in rice: identification and characterization of N-beta-D-glucopyranosyl indole-3-acetic acid and its conjugates

A search was made for conjugates of indole-3-acetic acid (IAA) in rice (Oryza sativa) using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) in order to elucidate unknown metabolic pathways for IAA. N-beta-d-Glucopyranosyl indole-3-acetic acid (IAA-N-Glc) was found in an alkaline hydrolysate of rice extract. A quantitative analysis of 3-week-old rice demonstrated that the total amount of IAA-N-Glc was equal to that of IAA. A LC-ESI-MS/MS-based analysis established that the major part of IAA-N-Glc was present as bound forms with aspartate and glutamate. Their levels were in good agreement with the total amount of IAA-N-Glc during the vegetative growth of rice. Further detailed analysis showed that both conjugates highly accumulated in the root. The free form of IAA-N-Glc accounted for 60% of the total in seeds but could not be detected in the vegetative tissue. An incorporation study using deuterium-labeled compounds showed that the amino acid conjugates of IAA-N-Glc were biosynthesized from IAA-amino acids. IAA-N-Glc and/or its conjugates were also found in extracts of Arabidopsis, Lotus japonicus, and maize, suggesting that N-glucosylation of indole can be the common metabolic pathway of IAA in plants.     

Putative roles of bacteria in the carbon and nitrogen cycles in a tropical peat swamp forest

Tropical peat swamp forests are waterlogged, acidic, anoxic and oligotrophic ecosystems. They are important terrestrial carbon pools that help mitigating global warming through carbon sequestration in peat. This study aimed at investigating putative roles of bacteria in the carbon and nitrogen cycles in North Selangor peat swamp forest, Malaysia. Whole-genome sequencing was performed on four bacterial isolates using Illumina NextSeq 500 to decipher their genetic information while Gen III Microplate (Biolog) was applied to verify carbon source utilization. The isolates were identified as Dyella sp. strain C9, Dyella sp. strain C11, Klebsiella sp. strain C31 and Paraburkholderia sp. strain C35. Both Dyella spp. and Paraburkholderia sp. strain C35 were likely novel species while Klebsiella sp. strain C31 was a different strain of the type species, Klebsiella pneumoniae. Both genomic and bioassay results suggested the involvement of the isolates in the degradation of lignocellulose, carbohydrates, sugar alcohols, organic acids and aromatic compounds. The isolates could potentially perform methanotrophy, which helps to mitigate methane emissions from tropical peatlands. In addition, the isolates also contained genes encoding enzymes for nitrite, nitrate and nitric oxide reduction, as well as dissimilatory nitrate reduction to ammonium, which retains the nitrogen in the ecosystems. The results generated insights into potential functions of bacteria in the energy production and nutrient cycling of tropical peatlands, which are essential for the sustainability of high biomass and biodiversity in these ecosystems.     

A common‐sense approach to tropical peat swamp forest restoration in Southeast Asia

Tropical peat swamp forests (TPSFs) are found mainly in Southeast Asia and especially Indonesia. A total of 61% were lost between 1990 and 2015 and 6% remained in a pristine condition by 2015. Tropical peat swamps store vast amounts of carbon in their peat, but peat degradation, through drainage and fire, leads to high greenhouse gas emissions. This is gaining much international attention and, with it, policy initiatives and funding for restoration from local to landscape scales are being promoted. Unfortunately, although there is a now strong desire and need for TPSF restoration, methods are lacking. Ecological understanding is still at an early stage, and, even more so, in its applied use. There is an imbalance between the activities of TPSF restoration and sound ecological application. Furthermore, while many activities are underway and knowledge is being gained, these techniques are yet to be published. This article has been written to provide a common‐sense, practical guide to tropical peatland forest restoration which summarizes what we know to date, while acknowledging the gaps in our understanding. Topics covered include species selection, land assessment, land selection, and appropriate nursery, transplanting, and monitoring methods. The authors make no apologies that in places this reads like a manual as, given the importance of tropical peatland recovery and the recent attention and funding opportunities available, it is essential we now provide techniques to restoration practitioners working on the ground, and a basic common‐sense approach must be the starting point.