Mesophyll conductance limits photosynthesis in fluctuating light under combined drought and heat stresses.

Drought and heat stresses usually occur concomitantly in nature, with increasing frequency and intensity of both stresses expected due to climate change. The synergistic agricultural impacts of these compound climate extremes are much greater than those of the individual stresses. However, the mechanisms by which drought and heat stresses separately and concomitantly affect dynamic photosynthesis have not been thoroughly assessed. To elucidate this, we used tomato (Solanum lycopersicum) seedlings to measure dynamic photosynthesis under individual and compound stresses of drought and heat. Individual drought and heat stresses limited dynamic photosynthesis at the stages of diffusional conductance to CO2 and biochemistry, respectively. However, the primary limiting factor for photosynthesis shifted to mesophyll conductance under the compound stresses. Compared with the control, photosynthetic carbon gain in fluctuating light decreased by 38%, 73%, and 114% under the individual drought, heat, and compound stresses, respectively. Therefore, compound stresses caused a greater reduction in photosynthetic carbon gain in fluctuating light conditions than individual stress. These findings highlight the importance of mitigating the effects of compound climate extremes on crop productivity by targeting mesophyll conductance and improving dynamic photosynthesis.

Complementary water and nutrient utilization of perianth structural units help maintain long floral lifespan in Dendrobium.

Most orchids have high ornamental value with long-lived flowers. However, the mechanisms by which orchids maintain floral longevity are poorly understood. Here, we hypothesized that floral longevity in Dendrobium is maintained by high resource investment and complementary water and nutrient utilization in different structural units of the perianth. To test this hypothesis, we determined which water- and nutrient-related traits are correlated with flower longevity in 23 Dendrobium species or cultivars, and examined variations of the related traits during flower development of one long-lived cultivar. We found that floral longevity was correlated with dry mass per unit area of perianths and total flower biomass, which indicates that maintaining floral longevity requires increased resource investment. During development of long-lived flowers, labella showed a high capacity for water storage and nutrient reutilization, which could partly remedy high water demand and biomass investment. Sepals and petals, in contrast, had stronger desiccation avoidance and higher metabolic activity with lower biomass investment. These findings indicate that Dendrobium flowers maintain longevity by complementary water and nutrient utilization strategies in the sepals, petals and labella, with labella consuming more water and nutrients to extend flower display, and sepals and petals using a more conservative strategy.

Biomass and Active Compounds Accumulation of the Medicinal Orchid Pleione bulbocodioides in Response to Light Intensity and Irrigation Frequency.

Pseudobulbs of Pleione species are widely used as traditional medicine in Asian countries, but the mechanism of active compound accumulation remains unclear. In the present study, we investigated the accumulation of biomass and three active compounds (dactylorhin A, militarine and batatasin III) of Pleione bulbocodioides in response to different light intensities and irrigation frequencies. We found that single high light (65 % of full sunlight) or drought stress (14-day irrigation interval) increased active compounds accumulation but the combined effect of these two treatments decreased the total content of these three active compounds. This decrease was due to the plants under combined stress having a significantly lower photosynthetic rate, leaf area and longevity, leading to a dramatic decrease in pseudobulb biomass. Among all treatments, the highest total content of active compounds was observed in plants subjected to the high light level with a high water level (3-day irrigation interval), and plants under medium light intensity (30 % of full sunlight) also had considerable content of active compounds accumulation. To balance the quality and quantity of Pleione pseudobulbs during artificial cultivation, 30∼65 % of full sunlight with the avoidance of drought stress is recommended. Our results suggest the accumulation of the three active compounds is significantly influenced by light intensity and irrigation frequency, which may contribute to the artificial cultivation and quality control of medicinal Pleione.

Selection and Validation of Reference Genes for Quantitative Real-Time PCR Analysis of Development and Tissue-Dependent Flower Color Formation in Cymbidium lowianum.

The development and tissue-dependent color formation of the horticultural plant results in various color pattern flowers. Anthocyanins and carotenoids contribute to the red and yellow colors, respectively. In this study, quantitative real-time polymerase chain reaction (qRT-PCR) is used to analyze the expression profiles of anthocyanin and carotenoids biosynthesis genes in Cymbidium lowianum (Rchb.f.) Rchb.f. Appropriate reference gene selection and validation are required before normalization of gene expression in qRT-PCR analysis. Thus, we firstly selected 12 candidate reference genes from transcriptome data, and used geNorm and Normfinder to evaluate their expression stability in lip (divided into abaxial and adaxial), petal, and sepal of the bud and flower of C. lowianum. Our results show that the two most stable reference genes in different tissues of C. lowianum bud and flower are EF1δ and 60S, the most unstable reference gene is 26S. The expression profiles of the CHS and BCH genes were similar to FPKM value profiles after normalization to the two most stable reference genes, EF1δ and 60S, with the upregulated CHS and BCH expression in flower stage, indicating that the ABP and CBP were activated across the stages of flower development. However, when the most unstable reference gene, 26S, was used to normalize the qRT-PCR data, the expression profiles of CHS and BCH differed from FPKM value profiles, indicating the necessity of selecting stable reference genes. Moreover, CHS and BCH expression was highest in the abaxial lip and adaxial lip, respectively, indicating that the ABP and CBP were activated in abaxial and adaxial lip, respectively, resulting in a presence of red or yellow segments in abaxial and adaxial lip. This study is the first to provide reference genes in C. lowianum, and also provide useful information for studies that aim to understand the molecular mechanisms of flower color formation in C. lowianum.

Leafless epiphytic orchids share Ceratobasidiaceae mycorrhizal fungi.

Some epiphytic orchids in the tribe Vandeae are characterized by extremely vestigial leaves (even leafless). Thus, their leaves provide only a small proportion of carbon required for their growth and development, while a large portion of carbon may need to be supplied by their roots and mycorrhizal fungi (MF). The MF richness and composition of leafless epiphytic orchids, which belong to numerous genera with diverse ecophysiologies and wide geographical ranges, remain poorly understood. In this study, we identified the MF communities of seven leafless epiphytic species from three orchid genera from up to 17 sites in China using high-throughput sequencing. Our analyses revealed that the leafless epiphytic orchids have a highly specialized association with Ceratobasidiaceae. Several fungal OTUs were found in three different orchid genera and have promoted germinations of Chiloschista and Phalaenopsis, which may have been caused by convergent evolution of leafless epiphytic orchids. Furthermore, the MF composition of Taeniophyllum glandulosum was significantly affected by collection site and host tree. Our study provides new insights into mycorrhizal associations of epiphytic orchids.

Chemical Constituents of Species in the Genus Pleione (Orchidaceae) and the Implications from Molecular Phylogeny.

Pleiones are popular ornamental orchids and different species of Pleione are long being used as traditional medicine in many Asian countries. However, previous chemical investigations of the genus Pleione are restricted to only a few species. In the present study, high performance liquid chromatography (HPLC) fingerprint of Pleione plants was established, which in particular, eight common peaks were confirmed in 16 species/hybrids. Three of the compounds corresponding to the chromatographic peaks were identified by electrospray ionization tandem mass spectrometry (ESI-tandem-MS). HPLC analysis confirmed the studied taxa shared most of chemical compounds but the content of chemical compounds was significantly different between species. Comparison of hierarchical clustering result with phylogenetic tree revealed that closely related species have higher similarities in chemical constituents. In consideration of low chemical similarity between spring-flowering and autumn-flowering species, we suggest a discrimination of these two groups during medicinal use of the genus Pleione. Species with a large pseudobulb and with high content of a certain compound should be given priority in future artificial cultivation and medicinal cultivar breeding. We hope our findings will contribute to the quality control and promote conservation of such endangered plant group.

The decline in photosynthetic rate upon transfer from high to low light is linked to the slow kinetics of chloroplast ATP synthase in Bletilla striata.

Upon a sudden transition from high to low light, the rate of CO2 assimilation (AN) in some plants first decreases to a low level before gradually becoming stable. However, the underlying mechanisms remain controversial. The activity of chloroplast ATP synthase (gH+) is usually depressed under high light when compared with low light. Therefore, we hypothesize that upon a sudden transfer from high to low light, the relatively low gH+ restricts ATP synthesis and thus causes a reduction in AN. To test this hypothesis, we measured gas exchange, chlorophyll fluorescence, P700 redox state, and electrochromic shift signals in Bletilla striata (Orchidaceae). After the transition from saturating to lower irradiance, AN and ETRII decreased first to a low level and then gradually increased to a stable value. Within the first seconds after transfer from high to low light, gH+ was maintained at low levels. During further exposure to low light, gH+ gradually increased to a stable value. Interestingly, a tight positive relationship was found between gH+ and ETRII. These results suggested that upon a sudden transition from high to low light, AN was restricted by gH+ at the step of ATP synthesis. Taken together, we propose that the decline in AN upon sudden transfer from high to low light is linked to the slow kinetics of chloroplast ATP synthase.

Processes controlling programmed cell death of root velamen radicum in an epiphytic orchid.

BACKGROUND AND AIMS: Development of the velamen radicum on the outer surface of the root epidermis is an important characteristic for water uptake and retention in some plant families, particularly epiphytic orchids, for survival under water-limited environments. Velamen radicum cells derive from the primary root meristem; however, following this development, velamen radicum cells die by incompletely understood processes of programmed cell death (PCD). METHODS: We combined the use of transmission electron microscopy, X-ray micro-tomography and transcriptome methods to characterize the major anatomical and molecular changes that occur during the development and death of velamen radicum cells of Cymbidium tracyanum, a typical epiphytic orchid, to determine how PCD occurs. KEY RESULTS: Typical changes of PCD in anatomy and gene expression were observed in the development of velamen radicum cells. During the initiation of PCD, we found that both cell and vacuole size increased, and several genes involved in brassinosteroid and ethylene pathways were upregulated. In the stage of secondary cell wall formation, significant anatomical changes included DNA degradation, cytoplasm thinning, organelle decrease, vacuole rupture and cell wall thickening. Changes were found in the expression of genes related to the biosynthesis of cellulose and lignin, which are instrumental in the formation of secondary cell walls, and are regulated by cytoskeleton-related factors and phenylalanine ammonia-lyase. In the final stage of PCD, cell autolysis was terminated from the outside to the inside of the velamen radicum. The regulation of genes related to autophagy, vacuolar processing enzyme, cysteine proteases and metacaspase was involved in the final execution of cell death and autolysis. CONCLUSIONS: Our results showed that the development of the root velamen radicum in an epiphytic orchid was controlled by the process of PCD, which included initiation of PCD, followed by formation of the secondary cell wall, and execution of autolysis following cell death.

Physiological and transcriptomic analysis highlight key metabolic pathways in relation to drought tolerance in Rhododendron delavayi.

Rhododendron delavayi is an alpine evergreen ornamental plant, but water shortage limits its growth and development in urban gardens. However, the adaptive mechanism of alpine evergreen rhododendrons to drought remains unclear. Here, a water control experiment was conducted to study the physiological and transcriptomic response of R. delavayi to drought. The drought treatment for 9 days decreased photosynthetic rate, induced accumulation of reactive oxygen species (ROS), and damaged chloroplast ultrastructure of R. delavayi. However, the photosynthetic rate quickly recovered to the level before treatment when the plants were re-watered. De novo assembly of RNA-Seq data generated 86,855 unigenes with an average length of 1870 bp. A total of 22,728 differentially expressed genes (DEGs) were identified between the control and drought plants. The expression of most DEGs related to photosynthesis were down-regulated during drought stress, and were up-regulated when the plants were re-watered, including the DEGs encoding subunits of light-harvesting chlorophyll-protein complex, photosystem II and photosystem I reaction center pigment-protein complexes, and photosynthetic electron transport. The expressions of many DEGs related to signal transduction, flavonoid biosynthesis and antioxidant activity were also significantly affected by drought stress. The results indicated that the response of R. delavayi to drought involved multiple physiological processes and metabolic pathways. Photosynthetic adjustment, ROS-scavenging system, abscisic acid and brassinosteroid signal transduction pathway may play important roles to improve drought tolerance of R. delavayi. Our findings provided valuable information for understanding the mechanisms of drought tolerance employed by Rhododendron species.

Comparative physiological and proteomic analyses reveal different adaptive strategies by Cymbidium sinense and C. tracyanum to drought.

MAIN CONCLUSION: A terrestrial orchid, Cymbidium sinense appears to utilizes "remedy strategy", while an epiphytic orchid, C. tracyanum , employs a "precaution strategy" to drought stress based on morphological, physiological and proteomic analysis. Drought condition influences plant growth and productivity. Although the mechanism by which plants adapt to this abiotic stress has been studied extensively, the water-adaptive strategies of epiphytes grown in water-limited habitats remain undefined. Here, root and leaf anatomies, dynamic changes in physiological and proteomic responses during periods of drought stress and recovery studied in an epiphytic orchid (Cymbidium tracyanum) and a terrestrial orchid (C. sinense) to investigate their strategies for coping with drought. Compared with C. sinense, C. tracyanum showed stronger drought-resistant adaptive characteristics to drought because its leaves had more negative water potential at turgor loss point and roots had higher proportion of velamen radicum thickness. Although both species demonstrated quick recovery of photosynthesis after stress treatment, they differed in physiological and proteomic responses. We detected and functionally characterized 103 differentially expressed proteins in C. sinense and 104 proteins in C. tracyanum. These proteins were mainly involved in carbon and energy metabolism, photosynthesis, and defense responses. The up-regulated expression of plastid fibrillin may have contributed to the marked accumulation of jasmonates only in stressed C. sinense, while ferredoxin-NADP reductase up-regulation was only found in C. tracyanum which possibly related to the stimulation of cyclic electron flow that is linked with photoprotection. These physiological and proteomic performances suggest distinct adaptive strategies to drought stress between C. sinense (remedy strategy) and C. tracyanum (precaution strategy). Our findings may help improve our understanding about the ecological adaptation of epiphytic orchids.

In vivo regulation of thylakoid proton motive force in immature leaves.

In chloroplast, proton motive force (pmf) is critical for ATP synthesis and photoprotection. To prevent photoinhibition of photosynthetic apparatus, proton gradient (ΔpH) across the thylakoid membranes needs to be built up to minimize the production of reactive oxygen species (ROS) in thylakoid membranes. However, the regulation of thylakoid pmf in immature leaves is little known. In this study, we compared photosynthetic electron sinks, P700 redox state, non-photochemical quenching (NPQ), and electrochromic shift (ECS) signal in immature and mature leaves of a cultivar of Camellia. The immature leaves displayed lower linear electron flow and cyclic electron flow, but higher levels of NPQ and P700 oxidation ratio under high light. Meanwhile, we found that pmf and ΔpH were higher in the immature leaves. Furthermore, the immature leaves showed significantly lower thylakoid proton conductivity than mature leaves. These results strongly indicated that immature leaves can build up enough ΔpH by modulating proton efflux from the lumenal side to the stromal side of thylakoid membranes, which is essential to prevent photoinhibition via thermal energy dissipation and photosynthetic control of electron transfer. This study highlights that the activity of chloroplast ATP synthase is a key safety valve for photoprotection in immature leaves.

Photoinhibition of photosystem I in Nephrolepis falciformis depends on reactive oxygen species generated in the chloroplast stroma.

We studied how high light causes photoinhibition of photosystem I (PSI) in the shade-demanding fern Nephrolepis falciformis, in an attempt to understand the mechanism of PSI photoinhibition under natural field conditions. Intact leaves were treated with constant high light and fluctuating light. Detached leaves were treated with constant high light in the presence and absence of methyl viologen (MV). Chlorophyll fluorescence and P700 signal were determined to estimate photoinhibition. PSI was highly oxidized under high light before treatments. N. falciformis showed significantly stronger photoinhibition of PSI and PSII under constant high light than fluctuating light. These results suggest that high levels of P700 oxidation ratio cannot prevent PSI photoinhibition under high light in N. falciformis. Furthermore, photoinhibition of PSI in N. falciformis was largely accelerated in the presence of MV that promotes the production of superoxide anion radicals in the chloroplast stroma by accepting electrons from PSI. From these results, we propose that photoinhibition of PSI in N. falciformis is mainly caused by superoxide radicals generated in the chloroplast stroma, which is different from the mechanism of PSI photoinhibition in Arabidopsis thaliana and spinach. Here, we provide some new insights into the PSI photoinhibition under natural field conditions.

Anatomical and diffusional determinants inside leaves explain the difference in photosynthetic capacity between Cypripedium and Paphiopedilum, Orchidaceae.

Comparing with other angiosperms, most members within the family Orchidaceae have lower photosynthetic capacities. However, the underlying mechanisms remain unclear. Cypripedium and Paphiopedilum are closely related phylogenetically in Orchidaceae, but their photosynthetic performances are different. We explored the roles of internal anatomy and diffusional conductance in determining photosynthesis in three Cypripedium and three Paphiopedilum species, and quantitatively analyzed their diffusional and biochemical limitations to photosynthesis. Paphiopedilum species showed lower light-saturated photosynthetic rate (A N), stomatal conductance (g s), and mesophyll conductance (g m) than Cypripedium species. A N was positively correlated with g s and g m. And yet, in both species A N was more strongly limited by g m than by biochemical factors or g s. The greater g s of Cypripedium was mainly affected by larger stomatal apparatus area and smaller pore depth, while the less g m of Paphiopedilum was determined by the reduced surface area of mesophyll cells and chloroplasts exposed to intercellular airspace per unit of leaf area, and much thicker cell wall thickness. These results suggest that leaf anatomical structure is the key factor affecting g m, which is largely responsible for the difference in photosynthetic capacity between those two genera. Our findings provide new insight into the photosynthetic physiology and functional diversification of orchids.

Superoxide generated in the chloroplast stroma causes photoinhibition of photosystem I in the shade-establishing tree species Psychotria henryi.

Our previous studies indicated that high light induced significant photoinhibition of photosystem I (PSI) in the shade-establishing tree species Psychotria henryi. However, the underlying mechanism has not been fully clarified. In the present study, in order to investigate the mechanism of PSI photoinhibition in P. henryi, we treated detached leaves with constant high light in the presence of methyl viologen (MV) or a soluble α-tocopherol analog, 2,2,5,7,8-pentamethyl-6-chromanol (PMC). We found that MV significantly depressed photochemical quantum yields in PSI and PSII when compared to PMC. On condition that no PSI photoinhibition happened, although cyclic electron flow (CEF) was abolished in the MV-treated samples, P700 oxidation ratio was maintain at higher levels than the PMC-treated samples. In the presence of PMC, PSI photoinhibition little changed but PSII photoinhibition was significantly alleviated. Importantly, PSI photoinhibition was largely accelerated in the presence of MV, which stimulates the production of superoxide and subsequently other reactive oxygen species at the chloroplast stroma by accepting electrons from PSI. Furthermore, MV largely aggravated PSII photoinhibition when compared to control. These results suggest that high P700 oxidation ratio cannot prevent PSI photoinhibition in P. henryi. Furthermore, the superoxide produced in the chloroplast stroma is critical for PSI photoinhibition in the higher plant P. henryi, which is opposite to the mechanism underlying PSI photoinhibition in Arabidopsis thaliana and spinach. These findings highlight a new mechanism of PSI photoinhibition in higher plants.

PSI photoinhibition is more related to electron transfer from PSII to PSI rather than PSI redox state in Psychotria rubra.

Although it has been believed that wild-type plants are capable of protecting photosystem I (PSI) under high light, our previous study indicates that PSI is sensitive to high light in the shade-established tree species Psychotria rubra. However, the underlying physiological mechanisms are unclear. In this study, we examined the roles of electron transfer from PSII to PSI and PSI redox state in PSI photoinhibition in P. rubra by treatments with lincomycin (Lin), diuron (DCMU), and methyl viologen (MV). After exposure to 2000 µmol photons m(-2) s(-1) for 2 h, PSI activity decreased by 35, 29, 3, and 49 % in samples treated with H2O, Lin, DCMU, and MV, respectively. Meanwhile, the MV-treated samples showed higher P700 oxidation ratio than the H2O-treated samples, suggesting the PSI photoinhibition under high light was accompanied by high levels of P700 oxidation ratio. PSI photoinhibition was alleviated in the DCMU-treated samples but was accelerated in the MV-treated samples, suggesting that PSI photoinhibition in P. rubra was mainly controlled by electron transfer from PSII to PSI. Taking together, PSI photoinhibition is more related to electron transfer from PSII to PSI rather than PSI redox state in P. rubra, which is different from the mechanisms of PSI photoinhibition in Arabidopsis thaliana and cucumber.

Insusceptibility of oxygen-evolving complex to high light in Betula platyphylla.

High mountain plants growing at high altitude have to regularly cope with high light and high UV radiation that can lead to photodamage of oxygen-evolving complex (OEC). However, the underlying mechanism of photoprotection for OEC in high mountain plants is unclear. Sun leaves of Betula platyphylla were used to examine whether cyclic electron flow (CEF) around photosystem I (PSI) plays an important role in photoprotection for OEC. Our results indicated that the value of ETRI/ETRII ratio significantly increased under high light. With increasing light intensity, non-photochemical quenching (NPQ) gradually increased, and the fraction of P700 that is oxidized in a given state gradually increased. These results indicated that CEF was significantly activated under high light. After treatment with a high light of 1600 µmol photons m(-2) s(-1) for 8 h, the OEC activity did not decline, but the maximum quantum yield of PSII (F v /F m ) ratio significantly decreased. These results suggested that CEF-dependent generation of proton gradient across thylakoid membrane protected OEC activity against high light. Furthermore, the stability of PSI activity during exposure to high light suggested that the high CEF activity in B. platyphylla played an important role in photoprotection for PSI activity.

Effects of temperature on leaf hydraulic architecture of tobacco plants.

MAIN CONCLUSION: Modifications in leaf anatomy of tobacco plants induced greater leaf water transport capacity, meeting greater transpirational demands and acclimating to warmer temperatures with a higher vapor pressure deficit. Temperature is one of the most important environmental factors affecting photosynthesis and growth of plants. However, it is not clear how it may alter leaf hydraulic architecture. We grew plants of tobacco (Nicotiana tabacum) 'k326' in separate glasshouse rooms set to different day/night temperature conditions: low (LT 24/18 °C), medium (MT 28/22 °C), or high (HT 32/26 °C). After 40 days of such treatment, their leaf anatomies, leaf hydraulics, photosynthetic rates, and instantaneous water-use efficiency (WUEi) were measured. Compared with those under LT, plants exposed to HT or MT conditions had significantly higher values for minor vein density (MVD), stomatal density (SD), leaf area, leaf hydraulic conductance (K leaf), and light-saturated photosynthetic rate (A sat), but lower values for leaf water potential (ψ l) and WUEi. However, those parameters did not differ significantly between HT and MT conditions. Correlation analyses demonstrated that SD and K leaf increased in parallel with MVD. Moreover, greater SD and K leaf were partially associated with accelerated stomatal conductance. And then stomatal conductance was positively correlated with A sat. Therefore, under well-watered, fertilized conditions, when relative humidity was optimal, changes in leaf anatomy seemed to facilitate the hydraulic acclimation to higher temperatures, meeting greater transpirational demands and contributing to the maintenance of great photosynthetic rates. Because transpiration rate increased more with temperature than photosynthetic rate, WUEi reduced under warmer temperatures. Our results indicate that the modifications of leaf hydraulic architecture are important anatomical and physiological strategies for tobacco plants acclimating to warmer temperatures under a higher vapor pressure deficit.

Effect of growth temperature on the electron flow for photorespiration in leaves of tobacco grown in the field.

Photorespiration has been indicated as an important mechanism for maintaining CO2 assimilation and alleviating photodamage under conditions of high light and low CO2 . We tested the hypothesis that plants grown under a high temperature had greater electron flow for photorespiration compared with those grown under a relative low temperature. Responses of photosynthetic electron flow and CO2 assimilation to incident light intensity and intercellular CO2 concentration were examined in leaves of tobacco cultivar 'k326'. Plants were cultivated at three sites with different ambient temperatures (Zhengzhou, Zunyi and Jiangchuan). Under high light, plants grown in Zhengzhou (with the highest growth temperature in the three sites) showed higher effective quantum yield of photosystem II and total electron flow through photosystem II than that in Zunyi and Jiangchuan. However, regardless of light intensity and intercellular CO2 status, there were no significant differences among sites in the photosynthetic CO2 assimilation rate or electron flow devoted to the carboxylation of ribulose-1,5-bisphosphate (RuBP). As a result, plants grown at high temperature showed higher electron flow devoted to oxygenation of RuBP than plants grown at low temperature. These results suggested that enhancement of electron flow for photorespiration is an important strategy in tobacco for acclimating to high growth temperature.

Differences in the stimulation of cyclic electron flow in two tropical ferns under water stress are related to leaf anatomy.

Cyclic electron flow (CEF) plays an important role in photoprotection for angiosperms under environmental stresses. However, ferns are more sensitive to drought and their water transport systems are not as efficient as those of angiosperms, it is unclear whether CEF also contributes to photoprotection in these plants. Using Microsorum punctatum and Paraleptochillus decurrens, we studied the electron fluxes through both photosystem I (PSI) and photosystem II (PSII) under water stress and their leaf anatomies. Our goal was to determine if CEF functions in the photoprotection of these ferns and, if so, whether CEF stimulation is related to leaf anatomy. Compared with P. decurrens, M. punctatum had thicker leaves and cuticles and higher water storage capacity, but lower stomatal density and slower rate of water loss. During induced drought, the decrease in leaf water potential (Ψ(leaf) ) was more pronounced in P. decurrens than in M. punctatum. For both species, the decline in Ψ(leaf) was associated with a lower effective PSII quantum yield, photochemical quantum yield of PSI and electron transport rate (ETR), whereas increases were found in the quantum yield of regulated energy dissipation, CEF and CEF/ETR(II) ratio. Values for CEF and the CEF/ETR(II) ratio peaked in M. punctatum at a light intensity of 500-600 µmol m(-2) s(-1) vs only 150-200 µmol m(-2) s(-1) in P. decurrens. Therefore, our results indicate that the stimulation of CEF in tropical ferns contributes to their photoprotection under water stress, and is related to their respective drought tolerance and leaf anatomy.

Metabolic and transcriptomic analyses elucidate a novel insight into the network for biosynthesis of carbohydrate and secondary metabolites in the stems of a medicinal orchid Dendrobium nobile.

Dendrobium nobile is an important medicinal and nutraceutical herb. Although the ingredients of D. nobile have been identified as polysaccharides, alkaloids, amino acids, flavonoids and bibenzyls, our understanding of the metabolic pathways that regulate the synthesis of these compounds is limited. Here, we used transcriptomic and metabolic analyses to elucidate the genes and metabolites involved in the biosynthesis of carbohydrate and several secondary metabolites in the stems of D. nobile. A total of 1005 metabolites and 31,745 genes were detected in the stems of D. nobile. The majority of these metabolites and genes were involved in the metabolism of carbohydrates (fructose, mannose, glucose, xylulose and starch), while some were involved in the metabolism of secondary metabolites (alkaloids, ß-tyrosine, ferulic acid, 4-hydroxybenzoate and chrysin). Our predicted regulatory network indicated that five genes (AROG, PYK, DXS, ACEE and HMGCR) might play vital roles in the transition from carbohydrate to alkaloid synthesis. Correlation analysis identified that six genes (ALDO, PMM, BGLX, EGLC, XYLB and GLGA) were involved in carbohydrate metabolism, and two genes (ADT and CYP73A) were involved in secondary metabolite biosynthesis. Our analyses also indicated that phosphoenol-pyruvate (PEP) was a crucial bridge that connected carbohydrate to alkaloid biosynthesis. The regulatory network between carbohydrate and secondary metabolite biosynthesis established will provide important insights into the regulation of metabolites and biological systems in Dendrobium species.