Changes in photosynthesis of alpine plant <Emphasis Type="Italic">Saussurea superba</Emphasis> during leaf expansion

The native alpine plant Saussurea superba is widely distributed in Qinghai–Tibetan Plateau regions. The leaves of S. superba grow in whorled rosettes, and are horizontally oriented to maximize sunlight exposure. Experiments were conducted in an alpine Kobresia humilis meadow near Haibei Alpine Meadow Ecosystem Research Station (37°29′–37°45′N, 101°12′–101°33′E; alt. 3200 m). Leaf growth, photosynthetic pigments and chlorophyll fluorescence parameters were measured in expanding leaves of S. superba. The results indicate that leaf area increased progressively from inner younger leaves to outside fully expanded ones, and then slightly decreased in nearly senescent leaves, due to early unfavorable environmental conditions, deviating from the ordinary growth pattern. The specific leaf area decreased before leaves were fully expanded, and the leaf thickness was largest in mature leaves. There were no significant changes in the content of chlorophylls (Chl) and carotenoids (Car), but the ratios of Chl a/b and Car/Chl declined after full expansion of the leaves. The variation of Chl a/b coincided well with changes in photochemical quenching (q _P) and the fraction of open PSII reaction centers (q _L). The maximum quantum efficiency of PSII photochemistry after 5 min dark relaxation (F _(v)/F _(m)) continuously increased from younger leaves to fully mature leaves, suggesting that mature leaves could recover more quickly from photoinhibition than younger leaves. The light-harvesting capacity was relatively steady during leaf expansion, as indicated by the maximum quantum efficiency of open PSII centers (\(F_{\text{v}}^{{\prime }}\)/\(F_{\text{m}}^{{\prime }}\)). UV-absorbing compounds could effectively screen harmful solar radiation, and are a main protection way on the photosynthetic apparatus. The decline of q _P and q _L during maturation, together with limitation of quantum efficiency of PSII reaction centers (L _(PFD)), shows a decrease of oxidation state of Q_A in PSII reaction centers under natural sunlight. Furthermore, light-induced (Φ _NPQ) and non-light-induced quenching (Φ _NO) were consistent with variation of L _(PFD). It is concluded that the leaves of S. superba could be classified into four functional groups: young, fully expanded, mature, and senescent. Quick recovery from photoinhibition was correlated with protection by screening pigments, and high level of light energy trapping was correlated with preservation of photosynthetic pigments. Increasing of Φ _NPQ and Φ _NO during leaves maturation indicates that both thermal dissipation of excessive excitation energy in safety and potential threat to photosynthetic apparatus were strengthened due to the declination of q _P and q _L, and enhancement of L _(PFD).     

Contrasting patterns of sun-red and shade-green leaves of <Emphasis Type="Italic">Buxus microphylla</Emphasis> in response to gradients of excess light during winter acclimation

Leaf reddening in overwintering evergreens largely restricts their application in landscapes and is generally triggered in response to excess light. To explore how leaves respond to excess light and examine the potential relevance of leaf reddening in this process, a comparative field study was conducted on the sun leaves (SUL), shade leaves (SHL) and three levels of artificially shaded sun leaves (SSUL) of Buxus microphylla ‘Wintergreen’. The seasonal changes in leaf colorations, chlorophyll (Chl) and carotenoid contents, leaf absorbance and chlorophyll fluorescence characteristics were investigated. The results showed that SUL upregulated Chl a/b with increased reductions in Chl b compared with Chl a, accumulated red pigments in the upper palisade mesophyll with reduced absorption in blue and red light but increased absorption in green light, and additionally, significantly downregulated photochemical activities through the sustained enhancement of energy dissipation in PSII antenna (ΦD) from fall to midwinter. In the SSUL, as the light intensity decreased, all of the above processes were mitigated except that the SSUL maintained constant absorptions in blue light region and whose levels were similar to those of the SUL and SHL. In contrast, the SHL maintained relatively high levels of Chl a and Chl b, remained completely green and showed regulated ΦD and ΦE (energy dissipation in PSII reaction centers) to maintain relatively high photochemical activity in the winter. We conclude that the sun leaves downregulate Chl contents to reduce the light absorption and simultaneously enhance sustained ΦD to dissipate most of the light energy, whereas shade leaves maintain relatively high Chl contents and demonstrate regulated proportions of ΦD and ΦE to match the extent to which the absorbed light can be utilized through photochemical reactions. The accumulated red pigments in sun phenotypes may provide a shading effect on Chls by directing energy to non-photosynthetic reaction centers in the blue light region where the absorption is offset by the reduced Chls.     

Functional Conservation of an <Emphasis Type="Italic">AGAMOUS</Emphasis> Orthologous Gene Controlling Reproductive Organ Development in the Gymnosperm Species <Emphasis Type="Italic">Taxus chinensis</Emphasis> var. <Emphasis Type="Italic">mairei</Emphasis>

Arabidopsis AGAMOUS (AG) has roles in specifying reproductive organ (stamens and carpels) identity, floral meristem determinacy, and repression of A-function. To investigate possible roles of AG orthologous genes in gymnosperm species and evolution of C function, we isolated and identified AG orthologous gene TcAG from Taxus chinensis var. mairei (family Taxaceae, order Coniferales), a member of the last divergant lineage from higher Conifer that sisters to Gnetales. Sequence alignment and phylogenetic analysis grouped TcAG into the gymnosperm AG lineage. TcAG was expressed in both developing male and female cones, but there was no expression in juvenile leaves. Ectopic expression of TcAG in an Arabidopsis ag mutant produced flowers with the third whorl petaloid stamen and fourth whorl normal carpel, but failed to convert first whorl sepals into carpeloid organs and second whorl petals into stamenoid organs. A 35S::TcAG transgenic Arabidopsis ag mutant had very early flowering, and produced a misshapen inflorescence with a shortened floral axis. Our results suggest that establishment of the complete C-function occurred gradually during AG lineage evolution even in gymnosperms.     

Oviposition by herbivorous insects induces changes in optical and mechanical properties of <Emphasis Type="Italic">Prunus avium</Emphasis> leaves

This study of animal–plant interaction focused on the impact of oviposition by an insect on the leaves of Prunus avium (cherries). We examined whether the oviposition by Caliroa cerasi affects leaf mechanical and spectral traits in P. avium. Three cultivars of P. avium were studied. Infested leaves had from 1 to 18 eggs and exhibited higher leaf dry mass per area (LMA) than leaves without eggs. Leaf dry weight and LMA were positively correlated with egg number per leaf. Infested leaves tended to have higher number of trichomes. Leaf thickness and material and structural resistance tended to increase in infested leaves. The reflectance across all wavelengths (500–700 nm) in leaves with larger number of eggs was higher compared to leaves without eggs. Photosynthetic performance was reduced and oxidative activity was increased in leaves with eggs. Extrafloral nectaries increased with increasing the number of eggs per leaf and thus play an important role in defense against herbivores by providing nectar rewards that attract their depredators. These responses to oviposition may be beneficial for the plants in terms of resistance to feeding larvae.     

Fine mapping of the chromosome 5B region carrying closely linked rust resistance genes <Emphasis Type="Italic">Yr47</Emphasis> and <Emphasis Type="Italic">Lr52</Emphasis> in wheat

Key message

Fine mapping of Yr47 and Lr52 in chromosome arm 5BS of wheat identified close linkage of the marker sun180 to both genes and its robustness for marker-assisted selection was demonstrated.

Abstract

The widely effective and genetically linked rust resistance genes Yr47 and Lr52 have previously been mapped in the short arm of chromosome 5B in two F₃ populations (Aus28183/Aus27229 and Aus28187/Aus27229). The Aus28183/Aus27229 F₃ population was advanced to generate an F₆ recombinant inbred line (RIL) population to identify markers closely linked with Yr47 and Lr52. Diverse genomic resources including flow-sorted chromosome survey sequence contigs representing the orthologous region in Brachypodium distachyon, the physical map of chromosome arm 5BS, expressed sequence tags (ESTs) located in the 5BS6-0.81-1.00 deletion bin and resistance gene analog contigs of chromosome arm 5BS were used to develop markers to saturate the target region. Selective genotyping was also performed using the iSelect 90 K Infinium wheat SNP assay. A set of SSR, STS, gene-based and SNP markers were developed and genotyped on the Aus28183/Aus27229 RIL population. Yr47 and Lr52 are genetically distinct genes that mapped 0.4 cM apart in the RIL population. The SSR marker sun180 co-segregated with Lr52 and mapped 0.4 cM distal to Yr47. In a high resolution mapping population of 600 F₂ genotypes Yr47 and Lr52 mapped 0.2 cM apart and marker sun180 was placed 0.4 cM distal to Lr52. The amplification of a different sun180 amplicon (195 bp) than that linked with Yr47 and Lr52 (200 bp) in 204 diverse wheat genotypes demonstrated its robustness for marker-assisted selection of these genes.

     

Sunfleck limits the small-scale distribution of endangered <Emphasis Type="Italic">Kingdonia uniflora</Emphasis> in the natural habitat of subalpine forest proved by its photosynthesis

The physiological effects of sunflecks on understory plants are poorly understood. Kingdonia uniflora is an endemic and endangered species in China, with a patchy distribution over much of its range. Sunflecks are reportedly the likely dominant factor in determining its patchy distribution. We studied the photosynthesis of K. uniflora in the field to test whether understory sunflecks result in photoinhibition and, thereby, potentially influence its patchy distribution. K. uniflora exhibited the low dark respiration rates, low light compensation points, and low light saturation points characteristic of shade-tolerant plants, allowing maintenance during the long periods of low understory light. Moreover, K. uniflora was able to regulate light energy utilization by non-photochemical quenching in low light. Gas exchange parameters were measured in six treatments (sunfleck-enriched, sunfleck-enriched with added saturation light, sunfleck-enriched with filtered ultraviolet-B (UV-B) radiation , sunfleck-limited, sunfleck-limited with added saturation light, and sunfleck-limited with filtered UV-B). The sunfleck-enriched treatment caused photoinhibition in K. uniflora, in part due to a UV-B-induced decrease in Pn. In addition, the application of simulated sunflecks indicated that K. uniflora leaves do not need continuous light. The photosynthetic responses of K. uniflora to sunflecks indicate that the sunflecks are a limiting factor in the small-scale distribution of K. uniflora.     

Dynamic changes in energy metabolism and electron transport of photosystem II in <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> infested by nymphs of <Emphasis Type="Italic">Bemisia tabaci</Emphasis> (Middle East-Asia Minor 1)

Bemisia tabaci Middle East-Asia Minor 1 (MEAM1) infestation adversely affected photosynthesis of host plants. In the current study, chlorophyll a fluorescence was measured to determine the effects of MEAM1 nymph infestation of tobacco local and systemic leaves on energy metabolism and electron transport of photosystemII(PSII). The results showed that the density of PSII reaction centres per excited cross section (CS) (RC/CS) of infested and systemic leaves was reduced at 14 and 20 days. In systemic leaves, the number of PSII closed reaction centres (1-qP) increased significantly at 14 and 20 days. Absorption flux per Q_A^? reducing PSII reaction centre (RC) (ABS/RC), trapped energy flux per RC (TRo/RC), and electron transport per RC (ETo/RC) of infested and systemic leaves increased with MEAM1 nymph infestation. The most obvious increase in absorption flux per CS (ABS/CSo) and trapped energy flux per CS (TRo/CSo) of infested and systemic leaves occurred at 14 days. MEAM1 nymph infestation significantly reduced the energy required for PSII Q_A to be completely reduced (Sm) in tobacco leaves. These results suggested that MEAM1 nymph infestation caused changes in light-harvesting antenna system and deactivation of the reaction centre, resulting in the reduction of photons absorbed by reaction centres per unit area. MEAM1 nymph infestation, particularly the 3rd instar nymphs, decreased light utilization ability and increased excess excitation energy in tobacco leaves. With MEAM1 nymph infestation, the relative electron transport capacity of the entire electron transport chain decreased, and more light energy was used to reduce Q_A.     

Genetic determination of tannins and herbivore resistance in <Emphasis Type="Italic">Quercus ilex</Emphasis>

Genetic variability of trees influences the chemical composition of tissues. This determines herbivore impact and, consequently, herbivore performance. We evaluated the independent effects of plant genotype and provenance on the tannin content of holm oak (Quercus ilex) and their consequences for herbivory and performance of gypsy moth (Lymantria dispar) larvae. Oak seedlings of 48 open-pollinated families from six populations were grown in a common garden in central Spain. Half the plants were subjected to defoliation by gypsy moth larvae and the other half were destructively sampled for chemical analysis. Tannin content of leaves did not differ significantly among populations but differed significantly among families. Estimates of heritability (h ²) and quantitative genetic differentiation among populations for tannin content (Q _ST) were 0.83 and 0.12, respectively. Defoliation was not related to the tannin content of plants nor to spine and trichome densities of leaves, although positive family–mean associations were observed between defoliation and both seed weight and plant height (P < 0.003). Among the oak populations, differential increase in larval weight gain with defoliation was observed. Leaf tannin content in Q. ilex is genetically controlled but does not influence defoliation or predict performance of the larvae. Different efficiencies of food utilisation depending on the oak genotypes indicate that other plant traits are influencing the feeding patterns and fitness of L. dispar and consequent population dynamics.     

Elimination of macro elements from inoculation and co-cultivation media enhances the efficiency of <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation in <Emphasis Type="Italic">Petunia</Emphasis>

In order to evaluate the effect of inoculation and co-cultivation media elements on transformation frequency in Petunia hybrida, modified MS media with different elements were tested on Alvan and Large Flower Alvan (LF Alvan), two local cultivars. Leaf explants of both cultivars were inoculated with Agrobacterium tumefaciens strain LBA4404 (pBI121) containing neomycin phosphotransferase (nptII) and an intron-containing β-glucuronidase (gus) genes. When medium lacking KH₂PO₄, NH₄NO₃, KNO₃, and CaCl₂ was used as inoculation and co-cultivation medium, a higher frequency of transformation for Alvan (22%) and LF Alvan (16%) was obtained. Kanamycin resistant plantlets were stained blue by GUS assay. Furthermore, polymerase chain reaction (PCR) analysis revealed the presence of both gus and nptII genes in all putative transformants. Finally, southern blot hybridization confirmed insertion of 1–4 copies of gus gene in transgenic plants.     

Genetics and fine mapping of a yellow-green leaf gene (<Emphasis Type="Italic">ygl</Emphasis>-<Emphasis Type="Italic">1</Emphasis>) in cabbage (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">capitata</Emphasis> L.)

Cabbage (Brassica oleracea var. capitata L.) is one of the most popular cultivated vegetables worldwide. Cabbage has rich phenotypic diversity, including plant height, head shape, head color, leaf shape and leaf color. Leaf color plays an important role in cabbage growth and development. At present, there are few reports on fine mapping of leaf color mutants in B. oleracea. In this study, a naturally occurring yellow-green leaf cabbage mutant (YL-1), derived from the self-pollinated progenies of the hybrid ‘Hosom’, was used for inheritance analysis and gene mapping. Segregation populations including F₂ and BC₁ were generated from the cross of two inbred lines, YL-1 and 01–20. Genetic analysis with the F₂ and BC₁ populations demonstrated that the yellow-green leaf color was controlled by a single recessive nuclear gene, ygl-1. Insertion–deletion (InDel) markers, designed based on the parental re-sequencing data, were used for the preliminary mapping with BSA (bulked segregant analysis) method. A genetic map constructed with 15 InDels indicated that ygl-1 was located on chromosome C01. The ygl-1 gene is flanked by InDel markers ID2 and M8, with genetic distances of 0.4 cM and 0.35 cM, respectively. The interval distance between two markers is 167 kb. Thus, it enables us to locate the ygl-1 gene for the first time in B. oleracea. This study lays the foundation for candidate gene prediction and ygl-1gene cloning.     

Reactive oxygen species (ROS) and antioxidative enzyme status in <Emphasis Type="Italic">Solanum lycopersicum</Emphasis> on priming with fluorescent <Emphasis Type="Italic">Pseudomonas</Emphasis> spp. against <Emphasis Type="Italic">Fusarium oxysporum</Emphasis>

In plants, ROS signaling and increase in activities of antioxidants are among defense responses. The present study describes the oxidative stress profiling in model host plant tomato (Solanum lycopersicum L.), during an invasion of the wilt pathogen Fusarium oxysporum f. sp. lycopersici with or without seed priming with Pseudomonas isolates M80, M96 and T109. Tomato seeds were primed with known Pseudomonas isolates M80, M96 and T109 and the forty-day- old plants were challenged with spores of F. oxysporum under greenhouse conditions. Leaf samples were collected at 0, 24, 48 72 and 96 h post fungal challenge and analysed for systemic level of oxidative stress parameters including total phenolics, proline, hydrogen peroxide, lipid peroxidation and enzymatic antioxidants. Disease incidence in the plants under greenhouse conditions was also calculated. Results revealed that priming with Pseudomonas isolates resulted in reduced oxidative stress in the host, during pathogen invasion. M80-priming showed highest antioxidative protection to the host plants during F. oxysporum invasion. The observed reduction in hydrogen peroxide and lipid peroxidation in primed plants was in agreement with the increased activities of the corresponding antioxidant enzymes. Greenhouse results showed that the highest wilt disease symptoms were with M80-priming followed by M96 and T109. The present study gives substantial evidences on the oxidative stress mitigation in response to Pseudomonas-priming on the model tomato-Fusarium interaction system.     

Effects of elevated atmospheric CO<Subscript>2</Subscript> and increased tidal flooding on leaf gas-exchange parameters of two common mangrove species: <Emphasis Type="Italic">Avicennia marina</Emphasis> and <Emphasis Type="Italic">Rhizophora stylosa</Emphasis>

In this study, we examined interactive effects of elevated atmospheric CO₂, concentrations, and increased tidal flooding on two mangroves species, Avicennia marina and Rhizophora stylosa. Leaf gas-exchange parameters (photosynthesis, transpiration rates, water-use efficiency, stomatal conductance, and dark respiration rates) were measured monthly on more than 1000 two-year-old seedlings grown in greenhouses for 1 year. In addition, stomatal density and light curve responses were determined at the end of the experiment. Under elevated CO₂ concentrations (800 ppm), the net photosynthetic rates were enhanced by more than 37% for A. marina and 45% for R. stylosa. This effect was more pronounced during the warm season, suggesting that an increase in global temperatures would further enhance the photosynthetic response of the considered species. Transpiration rates decreased by more than 15 and 8% for A. marina and R. stylosa, respectively. Consequently, water-use efficiency increased by 76% and 98% for A. marina and R. stylosa, respectively, for both species, which will improve drought resistance. These responses to elevated CO₂ were minimized (by 5%) with longer flooding duration. Consequently, future increases of atmospheric CO₂ may have a strong and positive effect on juveniles of A. marina and R. stylosa during the next century, which may not be suppressed by the augmentation of tidal flooding duration induced by sea-level rise. It is possible that this effect will enhance seedling dynamic by increasing photosynthesis, and therefore will facilitate their settlements in new area, extending the role of mangrove ecosystems in carbon sequestration and climate change mitigation.     

Phylogenetic,toxigenic and virulence profiles of <Emphasis Type="Italic">Alternaria</Emphasis> species causing leaf blight of tomato in Egypt

Species of Alternaria are serious plant pathogens, causing major losses on a wide range of crops. Leaf blight symptoms were observed on tomato leaves, and samples were collected from various regions. Isolation was done from symptomatic tomato leaves, and 15 representatives were selected from a collection of 65 isolates of Alternaria species. The virulence of Alternaria isolates was investigated on detached leaves (DL) and whole plants of tomato cv. Super strain B. A phylogenetic analysis was performed based on three partial gene regions, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the RNA polymerase second largest subunit (RPB2) and the Alternaria major allergen gene (Alt a 1). The potentiality of Alternaria isolates to produce toxins was also investigated on the basis of thin-layer chromatography (TLC). Our investigations revealed that Alternaria isolates showed different levels of virulence either on tomato plants or DL. Based on the phylogeny of three genes, Alternaria isolates encompassed two species of small-spored morphospecies: A. alternata (14 isolates) and A. arborescens (single isolate). The produced toxins varied among Alternaria isolates with tenuazonic acid (TeA) being the most abundant mycotoxin produced by most isolates. This study highlighted on other Alternaria species in Egypt that might represent a serious concern for tomato producers as causal agents of leaf blight over other species, i.e. A. solani.     

Mesophyll conductance to CO<Subscript>2</Subscript> in leaves of Siebold’s beech (<Emphasis Type="Italic">Fagus crenata</Emphasis>) seedlings under elevated ozone

Ozone is an air pollutant that negatively affects photosynthesis in woody plants. Previous studies suggested that ozone-induced reduction in photosynthetic rates is mainly attributable to a decrease of maximum carboxylation rate (V_cmax) and/or maximum electron transport rate (J_max) estimated from response of net photosynthetic rate (A) to intercellular CO₂ concentration (C_i) (A/C_i curve) assuming that mesophyll conductance for CO₂ diffusion (g_m) is infinite. Although it is known that C_i-based V_cmax and J_max are potentially influenced by g_m, its contribution to ozone responses in C_i-based V_cmax and J_max is still unclear. In the present study, therefore, we analysed photosynthetic processes including g_m in leaves of Siebold’s beech (Fagus crenata) seedlings grown under three levels of ozone (charcoal-filtered air or ozone at 1.0- or 1.5-times ambient concentration) for two growing seasons in 2016–2017. Leaf gas exchange and chlorophyll fluorescence were simultaneously measured in July and September of the second growing season. We determined the A, stomatal conductance to water vapor and g_m, and analysed A/C_i curve and A/C_c curve (C_c: chloroplast CO₂ concentration). We also determined the Rubisco and chlorophyll contents in leaves. In September, ozone significantly decreased C_i-based V_cmax. At the same time, ozone decreased g_m, whereas there was no significant effect of ozone on C_c-based V_cmax or the contents of Rubisco and chlorophyll in leaves. These results suggest that ozone-induced reduction in C_i-based V_cmax is a result of the decrease in g_m rather than in carboxylation capacity. The decrease in g_m by elevated ozone was offset by an increase in C_i, and C_c did not differ depending on ozone treatment. Since C_c-based V_cmax was also similar, A was not changed by elevated ozone. We conclude that g_m is an important factor for reduction in C_i-based V_cmax of Siebold’s beech under elevated ozone.     

Physiological alterations due to field salinity stress in melon (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.)

This study was aimed to assess physiological responses of melon (Cucumis melo L.) cultivars to salinity stress under field conditions. Seventeen melon cultivars including 16 widely distributed native and one exotic (‘Galia’) were subjected to 2-year (2014–2015) field salinity stress. Leaf relative water content (RWC), membrane stability index (MSI), pigments [chlorophyll a, b, total chlorophyll (TChl), carotenoid (Car) and their ratios], malondialdehyde (MDA), H₂O₂ content, proline content (Pro), total soluble sugar content (TSC), salinity tolerance and susceptibility indices as well as yield were evaluated. The results of combined analysis of variance showed significant genotypic variation for all the traits and significant effect of salinity stress on all the traits with the exception of Chla/Chlb and TChl/Car ratios. Overall, field salinity stress caused an increase in leaf MDA, H₂O₂, Chla, Chlb, TChl, Car, Pro and TSC and caused a reduction in leaf MSI and RWC as well as yield. The results of correlation coefficients showed that accumulation of osmolytes (proline and TSC) led to an increase in RWC and a decrease in MDA contents. In addition, the results of multiple regression analysis showed that leaf MDA, TSC, MSI and Chla contents were the most important predictors of yield justifying 72% total variation of yield under saline conditions. These results may highlight a dynamic interplay among biomarkers for lipid peroxidation (MDA), sugar osmolytes (TSC) and photosynthetic pigment (Chla) to maintain cell viability and cell wall integrity under salinity stress conditions in melon.     

Root-derived bicarbonate assimilation in response to variable water deficit in <Emphasis Type="Italic">Camptotheca acuminate</Emphasis> seedlings

Water deficit is one of the key factors that limits the carbon (C) assimilation and productivity of plants. The effect of variable water deficit on recently root-derived bicarbonate assimilation in Camptotheca acuminate seedlings was investigated. Three-month-old seedlings were subjected to three water regimes, well-watered (WW), moderate stress (MS), and severe stress (SS) induced by polyethyleneglycol, in conjunction with relatively high (H) and low (L) natural ¹³C-abundance of NaHCO₃-labeled treatments in hydroponics for 14 days. The δ¹³C of the newly expanded leaves in H were generally more enriched in heavy isotopes than were those in L, indicative of the involvement of bicarbonate in aboveground tissues. The C isotope fractionation of newly expanded leaves relative to air (?¹³C_air-leaves) ranged from 17.78 to 21.78‰ among the treatments. The ?¹³C_air-leaves under the MS and SS treatments in H were both more negative than was that in L. A linear regression between Ci/Ca and ?¹³C_air-leaves in both L and H were different from the theoretical regression. On the basis of the two end-member mixing model, the proportion of fixed CO₂ supplied from bicarbonate contributing to the total photosynthetically inorganic C assimilation were 10.34, 20.05 and 16.60% under the WW, MS, and SS treatments, respectively. These results indicated that the increase in water deficit decreased the atmospheric CO₂ gain but triggered a compensatory use of bicarbonate in C. acuminate seedlings.     

Two new species of <Emphasis Type="Italic">Gentianella</Emphasis> (Gentianaceae) from Peru

Two new species of Gentianella (Gentianaceae, Gentianeae, Swertiinae), G. grantii and G. wayqecha, are described from Departamento Cusco, Peru. These two species differ from other Peruvian species of Gentianella in a combination of stems 10–100 cm long; no rosette of basal leaves; cauline leaves 10–75 mm long; flowers in thyrses; and corollas 14–26 mm long, campanulate, and adaxially glabrous. The leaves of G. grantii consist of an appressed pseudopetiole and a spreading, narrowly elliptic-oblong to linear blade; the corollas are lavender; and the corolla lobes are 0.6–0.7× as long as the tube. Gentianella grantii is similar to G. lythroides, of Bolivia, but differs in having more closely spaced leaves and less deeply lobed corollas. The leaves of G. wayqecha are sessile, lanceolate to ovate; the corollas are rose-violet; and the corolla lobes are 0.75–1.35× as long as the tube. Gentianella wayqecha is similar to G. rapunculoides, of Colombia and Ecuador, and G. ruizii, of Peru, but differs from both in its adaxially glabrous corollas and from G. rapunculoides in its less deeply lobed corollas. Both G. grantii and G. wayqecha grow in moist habitats near tree line, and are known only from a area northeast of the city of Cusco.     

An improved method for the simultaneous determination of photosynthetic O2 evolution and CO2 consumption in Rhizophora mucronata leaves

The photosynthetic gas-exchange has been assessed traditionally either as O₂ evolution or CO₂ consumption. In this study, we used a liquid-phase O₂ electrode combined with CO₂ optodes to examine simultaneously photosynthesis in intact leaves of mangrove Rhizophora mucronata. We verified suitable conditions for leaf photosynthetic rates by assessing pH levels and NaHCO₃ concentrations and compared these to the gas-exchange method at various PAR levels. The photosynthetic rate in response to pH exhibited a similar pattern both for O₂ evolution and CO₂ consumption, and higher rates were associated with intermediate pH compared with low and high pH values. The net photosynthetic quotient (PQ) of R. mucronata leaves ranged from 1.04–1.28. The PQ values, which were never lesser than 1, suggested that photorespiration did not occur in R. mucronata leaves under aqueous conditions. The similar maximum photosynthetic rates suggested that all measurements had a high capacity to adjust the photosynthetic apparatus under a light saturation condition. The simultaneous measurements of O₂ evolution and CO₂ consumption using the Clark oxygen electrode polarographic sensor with the CO₂ optode sensor provided a simple, stable, and precise measurement of PQ under aqueous and saturated light conditions.