The combined effects of water and nitrogen on the relationship between a native hemiparasite and its invasive host.

Stem hemiparasites are dependent on their hosts for water and nitrogen. Most studies, however, have assessed the influence of one factor on parasite : host associations, thus limiting our mechanistic understanding of their performance in nature. We investigated the combined effects of water and nitrogen (N) availability on both a host (Ulex europaeus) and its parasite (Cassytha pubescens). Parasite infection significantly decreased host shoot biomass and shoot : root ratio more severely in high water than low water, irrespective of N supply. Parasite stem [N] was significantly higher in high water than low water treatments, regardless of N supply, but parasite biomass did not vary among treatments. Irrespective of water and N supply, infected plants had significantly lower total, root and nodule biomass, predawn and midday quantum yields, maximum electron transport rates, water potentials and nitrogen concentration [N]. Parasite δ13 C was significantly higher than that of the host. Our results suggested that stem hemiparasites can better extract resources from hosts when water availability is high, resulting in a greater impact on the host under these conditions. When hemiparasitic plants are being investigated as a biocontrol for invasive weeds, they may be more effective in wetter habitats than in drier ones.

It Is Hot in the Sun: Antarctic Mosses Have High Temperature Optima for Photosynthesis Despite Cold Climate.

The terrestrial flora of Antarctica's frozen continent is restricted to sparse ice-free areas and dominated by lichens and bryophytes. These plants frequently battle sub-zero temperatures, extreme winds and reduced water availability; all influencing their ability to survive and grow. Antarctic mosses, however, can have canopy temperatures well above air temperature. At midday, canopy temperatures can exceed 15°C, depending on moss turf water content. In this study, the optimum temperature of photosynthesis was determined for six Antarctic moss species: Bryum pseudotriquetrum, Ceratodon purpureus, Chorisodontium aciphyllum, Polytrichastrum alpinum, Sanionia uncinata, and Schistidium antarctici collected from King George Island (maritime Antarctica) and/or the Windmill Islands, East Antarctica. Both chlorophyll fluorescence and gas exchange showed maximum values of electron transport rate occurred at canopy temperatures higher than 20°C. The optimum temperature for both net assimilation of CO2 and photoprotective heat dissipation of three East Antarctic species was 20-30°C and at temperatures below 10°C, mesophyll conductance did not significantly differ from 0. Maximum mitochondrial respiration rates occurred at temperatures higher than 35°C and were lower by around 80% at 5°C. Despite the extreme cold conditions that Antarctic mosses face over winter, the photosynthetic apparatus appears optimised to warm temperatures. Our estimation of the total carbon balance suggests that survival in this cold environment may rely on a capacity to maximize photosynthesis for brief periods during summer and minimize respiratory carbon losses in cold conditions.

The impact of a native hemiparasite on a major invasive shrub is affected by host size at time of infection.

Many studies have investigated the effect of parasitic plants on their hosts; however, few have examined how parasite impact is affected by host size. In a glasshouse experiment, we investigated the impact of the Australian native hemiparasitic vine, Cassytha pubescens, on a major invasive shrub, Ulex europaeus, of different sizes. Infected plants had significantly lower total, shoot, and root biomass, but the parasite's impact was more severe on small than on large hosts. When infected, small but not large hosts had significantly lower nodule biomass. Irrespective of size, infection significantly decreased the host shoot/root ratio, pre-dawn and midday quantum yields, maximum electron transport rates, and carbon isotope composition, and the host nodule biomass per gram of root biomass significantly increased in response to infection. Infection did not affect host foliar nitrogen concentration or midday shoot water potential. Parasite biomass was significantly lower on small relative to large hosts, but was similar when expressed on a per gram of host total biomass basis. Parasite stem nitrogen, phosphorus, and potassium concentrations were significantly greater when C. pubescens was growing on small than on large hosts. Our results clearly show that C. pubescens strongly decreases performance of this major invasive shrub, especially when hosts are small. This suggests that C. pubescens could be used most effectively as a native biocontrol when deployed on smaller hosts.

A native parasitic plant affects the performance of an introduced host regardless of environmental variation across field sites.

Increasing evidence from glasshouse studies shows that native hemiparasitic plants can significantly impact the performance and growth of introduced host plants. We investigated the effect of the native Australian hemiparasite Cassytha pubescens R.Br. on the introduced shrub Ulex europaeus L. at three field sites in South Australia. Parasite infection significantly decreased midday PSII efficiency (ΦPSII) and the maximum electron transport rates (ETRmax) of U. europaeus across sites. The impact of C. pubescens on the photosynthetic performance of U. europaeus may have been caused by infected plants having significantly lower N and K, but higher Fe and Al than uninfected plants at all sites. Significant Al and Fe enrichment in infected plants may be possibly due to the parasite indirectly inducing rhizosphere acidification. At two sites, C. pubescens significantly affected host Fv/Fm, indicating chronic photoinhibition in response to infection. The impact of infection on Fv/Fm was greatest at the wettest site, in line with an experiment where C. pubescens had more impact under high water availability. At this site, infected plants also had the highest foliar Fe and Al. The C isotope (δ13C) of infected plants was significantly lower than that of uninfected plants at only one site. Unusually, the δ13C of the parasite was the same as or significantly higher than that of the hosts. There were no site effects on parasite Fv/Fm or ΦPSII; however, ETRmax and δ13C varied across sites. The results suggest that this native parasite has negative effects on U. europaeus in the field, as was found for glasshouse studies. The abundance of this introduced weed in Australia could be negatively affected by C. pubescens infection.

Does nitrogen affect the interaction between a native hemiparasite and its native or introduced leguminous hosts?

Associations between plants and nitrogen (N)-fixing rhizobia intensify with decreasing N supply and come at a carbon cost to the host. However, what additional impact parasitic plants have on their leguminous hosts' carbon budget in terms of effects on host physiology and growth is unknown. Under glasshouse conditions, Ulex europaeus and Acacia paradoxa either uninfected or infected with the hemiparasite Cassytha pubescens were supplied (high nitrogen (HN)) or not (low nitrogen (LN)) with extra N. The photosynthetic performance and growth of the association were measured. Cassytha pubescens significantly reduced the maximum electron transport rates and total biomass of U. europaeus but not those of A. paradoxa, regardless of N. Infection significantly decreased the root biomass of A. paradoxa only at LN, while the significant negative effect of infection on roots of U. europaeus was less severe at LN. Infection had a significant negative impact on host nodule biomass. Ulex europaeus supported significantly greater parasite biomass (also per unit host biomass) than A. paradoxa, regardless of N. We concluded that rhizobia do not influence the effect of a native parasite on overall growth of leguminous hosts. Our results suggest that C. pubescens will have a strong impact on U. europaeus but not A. paradoxa, regardless of N in the field.

High water availability increases the negative impact of a native hemiparasite on its non-native host.

Environmental factors alter the impacts of parasitic plants on their hosts. However, there have been no controlled studies on how water availability modulates stem hemiparasites' effects on hosts. A glasshouse experiment was conducted to investigate the association between the Australian native stem hemiparasite Cassytha pubescens and the introduced host Ulex europaeus under high (HW) and low (LW) water supply. Cassytha pubescens had a significant, negative effect on the total biomass of U. europaeus, which was more severe in HW than LW. Regardless of watering treatment, infection significantly decreased shoot and root biomass, nodule biomass, nodule biomass per unit root biomass, F v/F m, and nitrogen concentration of U. europaeus. Host spine sodium concentration significantly increased in response to infection in LW but not HW conditions. Host water potential was significantly higher in HW than in LW, which may have allowed the parasite to maintain higher stomatal conductances in HW. In support of this, the δ(13)C of the parasite was significantly lower in HW than in LW (and significantly higher than the host). C. pubescens also had significantly higher F v/F m and 66% higher biomass per unit host in the HW compared with the LW treatment. The data suggest that the enhanced performance of C. pubescens in HW resulted in higher parasite growth rates and thus a larger demand for resources from the host, leading to poorer host performance in HW compared with LW. C. pubescens should more negatively affect U. europaeus growth under wet conditions rather than under dry conditions in the field.

Native hemiparasite and light effects on photoprotection and photodamage in a native host.

Plants infected with hemiparasites often have lowered rates of photosynthesis, which could make them more susceptible to photodamage. However, it is also possible that infected plants increase their photoprotective capacity by changing their pigment content and/or engagement of the xanthophyll cycle. There are no published studies investigating infection effects on host pigment dynamics and how this relates to host susceptibility to photodamage whether in high (HL) or low light (LL). A glasshouse experiment was conducted where Leptospermum myrsinoides Schltdl. either uninfected or infected with Cassytha pubescens R.Br. was grown in HL or LL and pigment content of both host and parasite were assessed. Infection with C. pubescens significantly decreased all foliar pigment concentrations (except chlorophyll b) in L. myrsinoides in both HL and LL. Xanthophyll cycle (violaxanthin, antheraxanthin, zeaxanthin; VAZ) and chlorophyll (Chl) pigments decreased in parallel in response to infection, hence, VAZ/Chl of the host was unaffected by C. pubescens in either HL or LL. Pre-dawn and midday de-epoxidation state [(A+Z)/(V+A+Z)] of L. myrsinoides was also unaffected by infection in both HL and LL. Thus, L. myrsinoides infected with C. pubescens maintained similar photoprotective capacity per unit chlorophyll and engagement of the xanthophyll cycle as uninfected plants. Even though midday quantum yield (ΦPSII) of HL plants was affected by infection, pre-dawn maximum quantum yields (Fv/Fm) of hosts were the same as uninfected plants whether in HL or LL. This ability of L. myrsinoides to maintain photoprotective capacity/engagement when infected by C. pubescens thereby preventing photodamage could explain this host's tolerance to hemiparasite infection.

Do wide crowns in arid woodland trees reflect hydraulic limitation and reduction of self-shading?

In arid regions many tree species develop broad crowns. A number of hypotheses involve trade-offs between growth in height and horizontal spreading, but there is no explanation for the switch from vertical to horizontal growth during development. Using Acacia papyrocarpa Benth as a model, we measured tree height and crown shape across different sites and topographic positions. We also measured δ13C of phyllodes from crown tops and lateral spreading branches. Trees were significantly taller at the base of a hill, where water availability is typically greater, than on the adjacent steep hillslope. In contrast, δ13C from the treetops was not significantly different across this topographic gradient, despite variation in tree height. In addition, δ13C was higher at treetops than in lower, lateral branches. These observations are consistent with hydraulic limitation to tree height. The shape of mature and young crowns in open environments was not symmetrical. At all sites, branches were shortest, but tree crowns tallest, on south-facing (i.e. shadiest) aspect of crowns. This suggests that light limitation may also affect crown development. If upper branches become water-limited and lower branches light-limited, then middle lateral branches become the less-stressed part of the crown and may grow more, producing a broad crown.

Contemporary reliance on bicarbonate acquisition predicts increased growth of seagrass Amphibolis antarctica in a high-CO2 world.

Rising atmospheric CO2 is increasing the availability of dissolved CO2 in the ocean relative to HCO3 (-). Currently, many marine primary producers use HCO3 (-) for photosynthesis, but this is energetically costly. Increasing passive CO2 uptake relative to HCO3 (-) pathways could provide energy savings, leading to increased productivity and growth of marine plants. Inorganic carbon-uptake mechanisms in the seagrass Amphibolis antarctica were determined using the carbonic anhydrase inhibitor acetazolamide (AZ) and the buffer tris(hydroxymethyl)aminomethane (TRIS). Amphibolis antarctica seedlings were also maintained in current and forecasted CO2 concentrations to measure their physiology and growth. Photosynthesis of A. antarctica was significantly reduced by AZ and TRIS, indicating utilization of HCO3 (-)-uptake mechanisms. When acclimated plants were switched between CO2 treatments, the photosynthetic rate was dependent on measurement conditions but not growth conditions, indicating a dynamic response to changes in dissolved CO2 concentration, rather than lasting effects of acclimation. At forecast CO2 concentrations, seedlings had a greater maximum electron transport rate (1.4-fold), photosynthesis (2.1-fold), below-ground biomass (1.7-fold) and increase in leaf number (2-fold) relative to plants in the current CO2 concentration. The greater increase in photosynthesis (measured as O2 production) compared with the electron transport rate at forecasted CO2 concentration suggests that photosynthetic efficiency increased, possibly due to a decrease in photorespiration. Thus, it appears that the photosynthesis and growth of seagrasses reliant on energetically costly HCO3 (-) acquisition, such as A. antarctica, might increase at forecasted CO2 concentrations. Greater growth might enhance the future prosperity and rehabilitation of these important habitat-forming plants, which have experienced declines of global significance.

Genome of the long-living sacred lotus (Nelumbo nucifera Gaertn.).

BACKGROUND: Sacred lotus is a basal eudicot with agricultural, medicinal, cultural and religious importance. It was domesticated in Asia about 7,000 years ago, and cultivated for its rhizomes and seeds as a food crop. It is particularly noted for its 1,300-year seed longevity and exceptional water repellency, known as the lotus effect. The latter property is due to the nanoscopic closely packed protuberances of its self-cleaning leaf surface, which have been adapted for the manufacture of a self-cleaning industrial paint, Lotusan. RESULTS: The genome of the China Antique variety of the sacred lotus was sequenced with Illumina and 454 technologies, at respective depths of 101× and 5.2×. The final assembly has a contig N50 of 38.8 kbp and a scaffold N50 of 3.4 Mbp, and covers 86.5% of the estimated 929 Mbp total genome size. The genome notably lacks the paleo-triplication observed in other eudicots, but reveals a lineage-specific duplication. The genome has evidence of slow evolution, with a 30% slower nucleotide mutation rate than observed in grape. Comparisons of the available sequenced genomes suggest a minimum gene set for vascular plants of 4,223 genes. Strikingly, the sacred lotus has 16 COG2132 multi-copper oxidase family proteins with root-specific expression; these are involved in root meristem phosphate starvation, reflecting adaptation to limited nutrient availability in an aquatic environment. CONCLUSIONS: The slow nucleotide substitution rate makes the sacred lotus a better resource than the current standard, grape, for reconstructing the pan-eudicot genome, and should therefore accelerate comparative analysis between eudicots and monocots.

In the heat of the night--alternative pathway respiration drives thermogenesis in Philodendron bipinnatifidum.

• Philodendron bipinnatifidum inflorescences heat up to 42 °C and thermoregulate. We investigated whether they generate heat via the cytochrome oxidase pathway uncoupled by uncoupling proteins (pUCPs), or the alternative oxidase (AOX). • Contribution of AOX and pUCPs to heating in fertile (FM) and sterile (SM) male florets was determined using a combination of oxygen isotope discrimination, protein and substrate analyses. • Both FM and SM florets thermoregulated independently for up to 30 h ex planta. In both floret types, AOX contributed > 90% of respiratory flux during peak heating. The AOX protein increased fivefold with the onset of thermogenesis in both floret types, whereas pUCP remained low throughout development. These data indicate that AOX is primarily responsible for heating, despite FM and SM florets potentially using different substrates, carbohydrates or lipids, respectively. Measurements of discrimination between O2 isotopes in strongly respiring SM florets were affected by diffusion; however, this diffusional limitation was largely overcome using elevated O2. • The first in vivo respiratory flux measurements in an arum show AOX contributes the bulk of heating in P. bipinnatifidum. Fine-scale regulation of AOX activity is post-translational. We also demonstrate that elevated O2 can aid measurement of respiratory pathway fluxes in dense tissues.

Functional transition in the floral receptacle of the sacred lotus (Nelumbo nucifera): from thermogenesis to photosynthesis.

The receptacle of the sacred lotus is the main source of heat during the thermogenic stage of floral development. Following anthesis, it enlarges, greens and becomes a fully functional photosynthetic organ. We investigated development of photosynthetic traits during this unusual functional transition. There were two distinct phases of pigment accumulation in receptacles. Lutein and photoprotective xanthophyll cycle pigments accumulated first with 64 and 95% of the maximum, respectively, present before anthesis. Lutein epoxide comprised 32% of total carotenoids in yellow receptacles, but declined with development. By contrast, more than 85% of maximum total chlorophyll, ß-carotene and Rubisco were produced after anthesis, and were associated with significant increases in maximum electron transport rates (ETR) and photochemical efficiency (Fv/Fm). Leaves and mature receptacles had similar Rubisco content and ETRs (>200 µmol m-2 s-1), although total chlorophyll and total carotenoid contents of leaves were significantly higher than those of green receptacles. Receptacle δ13C before anthesis was similar to that of leaves; consistent with leaf photosynthesis being the source of C for these tissues. In contrast, mature receptacles had significantly lower δ13C than leaves, suggesting that 14-24% of C in mature receptacles is the result of refixation of respired CO2.

Synchronicity of thermogenic activity, alternative pathway respiratory flux, AOX protein content, and carbohydrates in receptacle tissues of sacred lotus during floral development.

The relationships between heat production, alternative oxidase (AOX) pathway flux, AOX protein, and carbohydrates during floral development in Nelumbo nucifera (Gaertn.) were investigated. Three distinct physiological phases were identified: pre-thermogenic, thermogenic, and post-thermogenic. The shift to thermogenic activity was associated with a rapid, 10-fold increase in AOX protein. Similarly, a rapid decrease in AOX protein occurred post-thermogenesis. This synchronicity between AOX protein and thermogenic activity contrasts with other thermogenic plants where AOX protein increases some days prior to heating. AOX protein in thermogenic receptacles was significantly higher than in post-thermogenic and leaf tissues. Stable oxygen isotope measurements confirmed that the increased respiratory flux supporting thermogenesis was largely via the AOX, with little or no contribution from the cytochrome oxidase pathway. During the thermogenic phase, no significant relationship was found between AOX protein content and either heating or AOX flux, suggesting that regulation is likely to be post-translational. Further, no evidence of substrate limitation was found; starch accumulated during the early stages of floral development, peaking in thermogenic receptacles, before declining by 89% in post-thermogenic receptacles. Whilst coarse regulation of AOX flux occurs via protein synthesis, the ability to thermoregulate probably involves precise regulation of AOX protein, most probably by effectors such as alpha-keto acids.

Mechanisms of thermoregulation in plants.

Endothermic heating of floral tissues and even thermoregulation is known to occur in a number of plant species across a wide taxonomic range. The mechanisms by which flowers heat, however, are only just beginning to be understood, and even less is known about how heating is regulated in response to changes in ambient temperature. We have recently demonstrated that the alternative pathway of respiration, in which the alternative oxidase (AOX) rather than cytochrome C (COX) acts as terminal electron acceptor, is responsible for heat generation in one thermoregulating species, the sacred lotus (Nelumbo nucifera). In the March issue of the Journal of Experimental Botany we further demonstrated that AOX-mediated heat production in this species is regulated at both the level of gene expression and also post-translationally. Similarly, AOX has also been implicated in heat production in other thermogenic species. In this addendum we discuss the central role of AOX in heat production and how post-translational mechanisms may provide the fine control necessary for thermoregulation.

Contribution of the alternative pathway to respiration during thermogenesis in flowers of the sacred lotus.

We report results from in vivo measurements, using oxygen isotope discrimination techniques, of fluxes through the alternative and cytochrome respiratory pathways in thermogenic plant tissue, the floral receptacle of the sacred lotus (Nelumbo nucifera). Fluxes through both pathways were measured in thermoregulating flowers undergoing varying degrees of thermogenesis in response to ambient temperature. Significant increases in alternative pathway flux were found in lotus receptacles with temperatures 16 degrees C to 20 degrees C above ambient, but not in those with lesser amounts of heating. Alternative pathway flux in the hottest receptacles was 75% of the total respiratory flux. In contrast, fluxes through the cytochrome pathway did not change significantly during thermogenesis. These data support the hypothesis that increased flux through the alternative pathway is responsible for heating in the lotus and that it is unlikely that uncoupling proteins, which would have produced increased fluxes through the cytochrome pathway, contribute significantly to heating in this tissue. Comparisons of actual flux, with capacity determined using inhibitors, suggested that the alternative pathway was operating at close to maximum capacity in heating tissues of lotus. However, in nonheating tissues the inhibitor data significantly overestimated the alternative pathway flux. This confirms that isotopic measurements are necessary for accurate determination of fluxes through the two pathways.

Does water status of Eucalyptus largiflorens (Myrtaceae) affect infection by the mistletoe Amyema miquelii (Loranthaceae)?

The impact of soil salinity and host water status on the host / parasite association between Eucalyptus largiflorens (F.Muell.) and Amyema miquelii (Lehm. ex Miq.) Tiegh. was investigated in a semi-arid floodplain environment in southern Australia. Water status of potential hosts (i.e. uninfected E. largiflorens) was assessed at a range of sites with different soil salinities and then compared with mistletoe infection at the same sites. Pre-dawn leaf water potentials (Ψ) of uninfected E. largiflorens declined with increasing salinity, while leaf δ13C values increased. The proportion of infected E. largiflorens at each site decreased significantly with increasing soil salinity. A significant relationship was found between the proportion of infected trees at each site and leaf δ13C values, but not pre-dawn Ψ, of potential hosts. The impact of mistletoes on water status of infected trees was also investigated. Among infected trees, we found no significant relationship between mistletoe volume and either pre-dawn Ψ or leaf δ13C values of hosts. However, there was a significant relationship between host midday leaf Ψ and mistletoe volume, with hosts exhibiting increased stress as mistletoe volume increased. The data suggest that increasing water and / or salinity stress make E. largiflorens a less suitable host for A. miquelii.