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Different intensities of high temperatures affect the growth of photosynthetic cells in nature. To elucidate the underlying mechanisms, we cultivated the unicellular green alga Chlamydomonas reinhardtii under highly controlled photobioreactor conditions and revealed systems-wide shared and unique responses to 24-hour moderate (35°C) and acute (40°C) high temperatures and subsequent recovery at 25°C. We identified previously overlooked unique elements in response to moderate high temperature. Heat at 35°C transiently arrested the cell cycle followed by partial synchronization, up-regulated transcripts/proteins involved in gluconeogenesis/glyoxylate-cycle for carbon uptake and promoted growth. But 40°C disrupted cell division and growth. Both high temperatures induced photoprotection, while 40°C distorted thylakoid/pyrenoid ultrastructure, affected the carbon concentrating mechanism, and decreased photosynthetic efficiency. We demonstrated increased transcript/protein correlation during both heat treatments and hypothesize reduced post-transcriptional regulation during heat may help efficiently coordinate thermotolerance mechanisms. During recovery after both heat treatments, especially 40°C, transcripts/proteins related to DNA synthesis increased while those involved in photosynthetic light reactions decreased. We propose down-regulating photosynthetic light reactions during DNA replication benefits cell cycle resumption by reducing ROS production. Our results provide potential targets to increase thermotolerance in algae and crops.

The unicellular green alga Chlamydomonas reinhardtii is an excellent model organism to investigate many essential cellular processes in photosynthetic eukaryotes. Two commonly used background strains of Chlamydomonas are CC-1690 and CC-5325. CC-1690, also called 21gr, has been used for the Chlamydomonas genome project and several transcriptome analyses. CC-5325 is the background strain for the Chlamydomonas Library Project (CLiP). Photosynthetic performance in CC-5325 has not been evaluated in comparison with CC-1690. Additionally, CC-5325 is often considered to be cell-wall deficient, although detailed analysis is missing. The circadian rhythms in CC-5325 are also unclear. To fill these knowledge gaps and facilitate the use of the CLiP mutant library for various screens, we performed phenotypic comparisons between CC-1690 and CC-5325. Our results showed that CC-5325 grew faster heterotrophically in dark and equally well in mixotrophic liquid medium as compared to CC-1690. CC-5325 had lower photosynthetic efficiency and was more heat-sensitive than CC-1690. Furthermore, CC-5325 had an intact cell wall which had comparable integrity to that in CC-1690 but appeared to have reduced thickness. Additionally, CC-5325 could perform phototaxis, but could not maintain a sustained circadian rhythm of phototaxis as CC1690 did. Finally, in comparison to CC-1690, CC-5325 had longer cilia in the medium with acetate but slower swimming speed in the medium without nitrogen and acetate. Our results will be useful for researchers in the Chlamydomonas community to choose suitable background strains for mutant analysis and employ the CLiP mutant library for genome-wide mutant screens under appropriate conditions, especially in the areas of photosynthesis, thermotolerance, cell wall, and circadian rhythms.

The unicellular green alga Chlamydomonas reinhardtii is an excellent model organism to investigate many essential cellular processes in photosynthetic eukaryotes. Two commonly used background strains of Chlamydomonas are CC-1690 and CC-5325. CC-1690, also called 21gr, has been used for the Chlamydomonas genome project and several transcriptome analyses. CC-5325 is the background strain for the Chlamydomonas Library Project (CLiP). Photosynthetic performance in CC-5325 has not been evaluated in comparison with CC-1690. Additionally, CC-5325 is often considered to be cell-wall deficient, although detailed analysis is missing. The circadian rhythms in CC-5325 are also unclear. To fill these knowledge gaps and facilitate the use of the CLiP mutant library for various screens, we performed phenotypic comparisons between CC-1690 and CC-5325. Our results showed that CC-5325 grew faster heterotrophically in dark and equally well in mixotrophic liquid medium as compared to CC-1690. CC-5325 had lower photosynthetic efficiency and was more sensitive to heat than CC-1690. Furthermore, CC-5325 had an intact cell wall with comparable integrity to that in CC-1690, though appears to have reduced thickness. Finally, CC-5325 could perform phototaxis, but could not maintain a sustained circadian rhythm of phototaxis as CC160 did. Our results will be useful for researchers in the Chlamydomonas community to choose suitable background strains for mutant analysis and employ the CLiP mutant library for genome-wide mutant screens under appropriate conditions, especially in the areas of photosynthesis, thermotolerance, cell wall, and circadian rhythms.

The unicellular green alga

A striking feature of plant ecology is the ability of plants to detect and respond to environmental cues such as herbivore-induced plant volatiles (HIPVs) by priming or directly activating defenses against future herbivores. However, whether seeds also respond to compounds that are common constituents of HIPV blends and initiate future plant resistance is unknown. Considering that seeds depend on other environmental cues to determine basic survival traits such as germination timing, we predicted that seeds exposed to synthetic constituents of HIPV blends would generate well-defended plants. We investigated the effect of seed exposure to common volatiles on growth, reproduction, and resistance characteristics in the model plants Arabidopsis thaliana and Medicago truncatula using herbivores from two feeding guilds. After seed scarification and vernalization, we treated seeds with one of seven different plant-derived volatile compounds for 24 h. Seeds were then germinated and the resulting plants were assayed for growth, herbivore resistance, and expression of inducible defense genes. Of all the synthetic volatiles tested, indole specifically reduced both beet armyworm growth on A. thaliana and pea aphid fecundity on M. truncatula. The induction of defense genes was not affected by seed exposure to indole in either plant species, indicating that activation of direct resistance rather than inducible resistance is the mechanism by which seed priming operates. Moreover, neither plant species showed any negative effect of seed exposure to any synthetic volatile on vegetative and reproductive growth. Rather, M. truncatula plants derived from seeds exposed to (Z)-3-hexanol and (Z)-3-hexenyl acetate grew larger compared to controls. Our results indicate that seeds are sensitive to specific volatiles in ways that enhance resistance profiles with no apparent costs in terms of growth. Seed priming by HIPVs may represent a novel ecological mechanism of plant-to-plant interactions, with broad potential applications in agriculture and seed conservation.

Different intensities of high temperatures affect the growth of photosynthetic cells in nature. To elucidate the underlying mechanisms, we cultivated the unicellular green alga Chlamydomonas reinhardtii under highly controlled photobioreactor conditions and revealed systems-wide shared and unique responses to 24-hour moderate (35°C) and acute (40°C) high temperatures and subsequent recovery at 25°C. We identified previously overlooked unique elements in response to moderate high temperature. Heat at 35°C transiently arrested the cell cycle followed by partial synchronization, up-regulated transcripts/proteins involved in gluconeogenesis/glyoxylate-cycle for carbon uptake, promoted growth, and increased starch accumulation. Heat at 40°C arrested the cell cycle, inhibited growth, resulting in carbon uptake over usage and increased starch accumulation. Both high temperatures induced photoprotection, while 40°C decreased photosynthetic efficiency, distorted thylakoid/pyrenoid ultrastructure, and affected the carbon concentrating mechanism. We demonstrated increased transcript/protein correlation during both heat treatments, suggesting reduced post-transcriptional regulation during heat may help coordinate heat tolerance activities efficiently. During recovery after both heat treatments, transcripts/proteins related to DNA synthesis increased while those involved in photosynthetic light reactions decreased. We propose down-regulating photosynthetic light reactions during DNA replication benefits cell cycle resumption by reducing ROS production. Our results provide potential targets to increase thermotolerance in algae and crops.

Ozone and wounding are key abiotic factors but, their interactive effects on temporal changes in terpene synthase gene expression and emission responses are poorly understood. Here, we applied combined acute ozone and wounding stresses to the constitutive isoprenoid-emitter Eucalyptus globulus and studied how isoprene, 1,8-cineole, and isoledene synthase genes were regulated, and how the gene expression was associated with temporal changes in photosynthetic characteristics, product emission rates, and stomatal ozone uptake through recovery phase. Photosynthetic characteristics and emission rate of isoprene, 1,8-cineole, and isoledene were synergistically altered, while three TPS gene expressions were antagonistically altered by combined stress applications. A time-delay analysis indicated that the best correspondences between gene expression and product emission rates were observed for 0 h time-shift for wounding and 0-2 h time-shifts for separate ozone, and combined ozone and wounding treatments. The best correspondence between ozone uptake and gene expression was observed for 0-4 h time-shifts for separate ozone and combined ozone and wounding treatments. Overall, this study demonstrated that expression profiles of isoprene, the monoterpene 1,8-cineole, and the sesquiterpene isoledene synthase genes differentially influenced their corresponding product emissions for separate and combined ozone and wounding treatments through recovery.

Both ozone and wounding constitute two key abiotic stress factors, but their interactive effects on plant constitutive and stress-elicited volatile (VOC) emissions are poorly understood. Furthermore, the information on time-dependent modifications in VOC release during recovery from a combined stress is very limited. We studied the modifications in photosynthetic characteristics and constitutive and stress-induced volatile emissions in response to single and combined applications of acute ozone (4, 5, and 6 ppm) and wounding treatments through recovery (0.5-75 h) in a constitutive isoprene and mono- and sesquiterpene emitter Eucalyptus globulus. Overall, the photosynthetic characteristics were surprisingly resistant to all ozone and wounding treatments. Constitutive isoprene emissions were strongly upregulated by ozone and combined ozone and wounding treatments and remained high through recovery phase, but wounding applied alone reduced isoprene emission. All stress treatments enhanced emissions of lipoxygenase pathway volatiles (LOX), mono- and sesquiterpenes, saturated aldehydes (C7-C10), benzenoids, and geranylgeranyl diphosphate (GGDP) pathway volatiles. Once elicited, GGDP volatile, saturated aldehyde and benzenoid emissions remained high through the recovery period. In contrast, LOX emissions, and total mono- and sesquiterpene emissions decreased through recovery period. However, secondary rises in total sesquiterpene emissions at 75 h and in total monoterpenes at 25-50 h were observed. Overall, acute ozone and wounding treatments synergistically altered gas exchange characteristics and stress volatile emissions. Through the treatments and recovery period, stomatal ozone uptake rate and volatile emission rates were poorly correlated, reflecting possible ozone-scavenging effect of volatiles and thus, reduction of effective ozone dose and elicitation of induced defense by the acute ozone concentrations applied. These results underscore the important role of interactive stresses on both constitutive and induced volatile emission responses.

Ozone-driven isoprenoid emissions scaled with surface ozone uptake, indicating that isoprenoids were released from trichomes. Greater non-stomatal ozone uptake due to ozone detoxification on the leaf surface underlies greater ozone tolerance., Ozone is a strong oxidant and a key stress elicitor. The immediate and longer term impacts of ozone are poorly understood in species with emission of both de novo synthesized and stored volatiles, such a tobacco (Nicotiana tabacum), which has terpene-containing glandular trichomes on the leaf surface. In this study, we exposed N. tabacum ‘Wisconsin’ leaves to acute ozone doses of 0 (control), 400, 600, 800, and 1000 ppb for 30 min and studied the effects of ozone exposure on ozone uptake, gas-exchange characteristics, and emissions of lipoxygenase pathway volatiles, monoterpenes, and sesquiterpenes. Foliage emissions of lipoxygenase pathway volatiles were quantitatively related to the severity of ozone exposure, but the stress dose vs. emission relationship was weaker for terpenoids. Analysis of leaf terpene content and composition indicated that several monoterpenes and sesquiterpenes were not stored in leaves and were synthesized de novo upon ozone exposure. The highest degree of elicitation for each compound was observed immediately after ozone treatment and it declined considerably during recovery. Leaf ozone uptake was dominated by non-stomatal deposition, and the emissions of total lipoxygenase pathway volatiles and mono- and sesquiterpenes were positively correlated with non-stomatal ozone deposition. Overall, this study demonstrates remarkably high ozone resistance of the studied tobacco cultivar and indicates that ozone’s effects on volatile emissions primarily reflect modifications in the release of stored volatiles and reaction of ozone with the leaf surface structure.

Plants frequently experience heat ramps of various severities, but how and to what degree plant metabolic activity recovers from mild and severe heat stress is poorly understood. In this study, we exposed the constitutive terpene emitter, Solanum. lycopersicum leaves to mild (37 and 41 °C), moderate (46 °C) and severe (49 °C) heat ramps of 5 min. and monitored foliage photosynthetic activity, lipoxygenase pathway volatile (LOX), and mono- and sesquiterpene emissions and expression of two terpene synthase genes, β-phellandrene synthase and (E)-β-caryophyllene/α-humulene synthase, through a 24 h recovery period upon return to pre-stress conditions. Leaf monoterpene emissions were dominated by β-phellandrene and sesquiterpene emissions by (E)-β-caryophyllene, and thus, these two terpene synthase genes were representative for the two volatile terpene classes. Photosynthetic characteristics partly recovered under moderate heat stress, and very limited recovery was observed under severe stress. All stress treatments resulted in elicitation of LOX emissions that declined during recovery. Enhanced mono- and sesquiterpene emissions were observed immediately after the heat treatment, but the emissions decreased even to below the control treatment during recovery between 2-10 h, and raised again by 24 h. The expression of β-phellandrene and (E)-β-caryophyllene synthase genes decreased between 2-10 h after heat stress, and recovered to pre-stress level in mild heat stress treatment by 24 h. Overall, this study demonstrates a highly sensitive heat response of terpenoid synthesis that is mainly controlled by gene level responses under mild stress, while severe stress leads to non-recoverable declines in foliage physiological and gene expression activities.

Silver birch (Betula pendula) is a pioneer boreal tree that can be induced to flower within 1 year. Its rapid life cycle, small (440-Mb) genome, and advanced germplasm resources make birch an attractive model for forest biotechnology. We assembled and chromosomally anchored the nuclear genome of an inbred B. pendula individual. Gene duplicates from the paleohexaploid event were enriched for transcriptional regulation, whereas tandem duplicates were overrepresented by environmental responses. Population resequencing of 80 individuals showed effective population size crashes at major points of climatic upheaval. Selective sweeps were enriched among polyploid duplicates encoding key developmental and physiological triggering functions, suggesting that local adaptation has tuned the timing of and cross-talk between fundamental plant processes. Variation around the tightly-linked light response genes PHYC and FRS10 correlated with latitude and longitude and temperature, and with precipitation for PHYC. Similar associations characterized the growth-promoting cytokinin response regulator ARR1, and the wood development genes KAK and MED5A.

Heat stress induces secondary metabolic changes in plants, channeling photosynthetic carbon and energy, away from primary metabolic processes, including, growth. Use of ACC (1-aminocyclopropane-1-carboxylate) deaminase containing plant growth promoting bacteria (PGPB) in conferring heat resistance in plants and the role of PGPB, in altering net carbon assimilation, constitutive and stress volatile emissions has not been studied yet. We exposed leaves of Eucalyptus grandis inoculated and non-inoculated with PGPB Brevibacterium linens RS16 to two levels of heat stress (37 °C and 41 °C for 5 min) and quantified temporal changes in foliage photosynthetic characteristics and volatile emission rates at 0.5 h, day 1 and day 5 after the stress application. Heat stress resulted in immediate reductions in dark-adapted photosystem II (PSII) quantum yield (Fv/Fm), net assimilation rate (A), stomatal conductance to water vapor (gs), and enhancement of stress volatile emissions, including enhanced emissions of green leaf volatiles (GLV), mono- and sesquiterpenes, light weight oxygenated volatile organic compounds (LOC), geranyl-geranyl diphosphate pathway volatiles (GGDP), saturated aldehydes, and benzenoids, with partial recovery by day 5. Changes in stress-induced volatiles were always less in leaves inoculated with B. linens RS16. However, net assimilation rate was enhanced by bacterial inoculation only in the 37 °C treatment and overall reduction of isoprene emissions was observed in bacterially-treated leaves. Principal component analysis (PCA), correlation analysis and partial least squares discriminant analysis (PLS-DA) indicated that different stress applications influenced specific volatile organic compounds. In addition, changes in the expression analysis of heat shock protein 70 gene (DnaK) gene in B. linens RS16 upon exposure to higher temperatures further indicated that B. linens RS16 has developed its own heat resistance mechanism to survive under higher temperature regimes. Taken together, this study demonstrates that foliar application of ACC deaminase containing PGPB can ameliorate heat stress effects in realistic biological settings.

Inoculation of endophytic Methylobacterium oryzae CBMB20 in salt-stressed rice plants improves photosynthesis and reduces stress volatile emissions due to mellowing of ethylene-dependent responses and activating vacuolar H+-ATPase. The objective of this study was to analyze the impact of ACC (1-aminocyclopropane-1-carboxylate) deaminase-producing Methylobacterium oryzae CBMB20 in acclimation of plant to salt stress by controlling photosynthetic characteristics and volatile emission in salt-sensitive (IR29) and moderately salt-resistant (FL478) rice (Oryza sativa L.) cultivars. Saline levels of 50 mM and 100 mM NaCl with and without bacteria inoculation were applied, and the temporal changes in stress response and salinity resistance were assessed by monitoring photosynthetic characteristics, ACC accumulation, ACC oxidase activity (ACO), vacuolar H+ ATPase activity, and volatile organic compound (VOC) emissions. Salt stress considerably reduced photosynthetic rate, stomatal conductance, PSII efficiency and vacuolar H+ ATPase activity, but it increased ACC accumulation, ACO activity, green leaf volatiles, mono- and sesquiterpenes, and other stress volatiles. These responses were enhanced with increasing salt stress and time. However, rice cultivars treated with CBMB20 showed improved plant vacuolar H+ ATPase activity, photosynthetic characteristics and decreased ACC accumulation, ACO activity and VOC emission. The bacteria-dependent changes were greater in the IR29 cultivar. These results indicate that decreasing photosynthesis and vacuolar H+ ATPase activity rates and increasing VOC emission rates in response to high-salinity stress were effectively mitigated by M. oryzae CBMB20 inoculation.

The emission of volatiles in response to salt stress in rice cultivars has not been studied much to date. Studies addressing the regulation of stress induced volatile emission by halotolerant plant growth promoting bacteria containing ACC (1-aminocyclopropane-1-carboxylate) deaminase are also limited. The objective of the present study was to investigate the salt alleviation potential of bacteria by regulating photosynthetic characteristics and volatile emissions in rice cultivars, and to compare the effects of the bacteria inoculation and salt responses between two rice genotypes. The interactive effects of soil salinity (0, 50, and 100 mM NaCl) and inoculation with Brevibacterium linens RS16 on ACC accumulation, ACC oxidase activity, carbon assimilation and stress volatile emissions after stress application were studied in the moderately salt resistant (FL478) and the salt-sensitive (IR29) rice (Oryza sativa L.) cultivars. It was observed that salt stress reduced foliage photosynthetic rate, but induced foliage ACC accumulation, foliage ACC oxidase activity, and the emissions of all the major classes of volatile organic compounds (VOCs) including the lipoxygenase pathway volatiles, light-weight oxygenated volatiles, long-chained saturated aldehydes, benzenoids, geranylgeranyl diphosphate pathway products, and mono- and sesquiterpenes. All these characteristics scaled up quantitatively with increasing salt stress. The effects of salt stress were more pronounced in the salt-sensitive genotype IR29 compared to the moderately salt resistant FL478 genotype. However, the bacterial inoculation significantly enhanced photosynthesis, and decreased ACC accumulation and the ACC oxidase activity, and VOC emissions both in control and salt-treated plants. Taken together, these results suggested that the ACC deaminase-containing Brevibacterium linens RS16 reduces the temporal regulation of VOC emissions and increases the plant physiological activity by reducing the availability of ethylene precursor ACC and the ACC oxidase activity under salt stress.

Plants frequently experience heat ramps of various severities, but how and to what degree plant metabolic activity recovers from mild and severe heat stress is poorly understood. In this study, we exposed the constitutive terpene emitter, Solanum. lycopersicum leaves to mild (37 and 41 °C), moderate (46 °C) and severe (49 °C) heat ramps of 5 min. and monitored foliage photosynthetic activity, lipoxygenase pathway volatile (LOX), and mono- and sesquiterpene emissions and expression of two terpene synthase genes, β-phellandrene synthase and (E)-β-caryophyllene/α-humulene synthase, through a 24 h recovery period upon return to pre-stress conditions. Leaf monoterpene emissions were dominated by β-phellandrene and sesquiterpene emissions by (E)-β-caryophyllene, and thus, these two terpene synthase genes were representative for the two volatile terpene classes. Photosynthetic characteristics partly recovered under moderate heat stress, and very limited recovery was observed under severe stress. All stress treatments resulted in elicitation of LOX emissions that declined during recovery. Enhanced mono- and sesquiterpene emissions were observed immediately after the heat treatment, but the emissions decreased even to below the control treatment during recovery between 2-10 h, and raised again by 24 h. The expression of β-phellandrene and (E)-β-caryophyllene synthase genes decreased between 2-10 h after heat stress, and recovered to pre-stress level in mild heat stress treatment by 24 h. Overall, this study demonstrates a highly sensitive heat response of terpenoid synthesis that is mainly controlled by gene level responses under mild stress, while severe stress leads to non-recoverable declines in foliage physiological and gene expression activities.

Genetic variation is an important attribute of forest tree populations enabling them to adapt to spatial and temporal variations in environmental conditions. In particular, Scots pine (Pinus sylvestris L.) has an exceptionally broad area of dispersal covering different climates and soil conditions, but the genetic variability in extreme conditions has not been studied. We hypothesized that the genetic variability of P. sylvestris is enhanced at its soil fertility limit in bogs where stunted trees form a sparse canopy with reduced light competition, but in highly turbulent conditions generating ideal conditions for distant pollen dispersal. A total of 180 individuals were studied from three bog populations using five nuclear (nSSR) and five chloroplastic (cpSSR) polymorphic microsatellite loci (simple sequence repeats, SSR). According to both marker systems, high and similar level of genetic diversity (ca. 99 % within the populations and 1 % among the populations) was observed for nuclear (F ST = 0.007 and R ST = 0.022) and chloroplast (F ST = 0.015 and R ST = 0.023) genomes. Despite the low genetic differentiation among the populations, there was evidence of geographic genetic differentiation in the chloroplast genomes, suggesting that isolation by distance might be a possible mechanism shaping the present distribution of genetic variability. In addition, significant but low spatial genetic structure along habitat wetness gradient was found in nuclear genomes in one site. Significant cyto-nuclear linkage disequilibrium was shown between one of the cpSSR loci to all nSSR loci. The results demonstrate a huge within-population genetic variability in these sites and underscore the importance of pollen gene flow in homogenizing populations on these geographic scales.

Silver birch (Betula pendula) is a pioneer boreal tree that can be induced to flower within 1 year. Its rapid life cycle, small (440-Mb) genome, and advanced germplasm resources make birch an attractive model for forest biotechnology. We assembled and chromosomally anchored the nuclear genome of an inbred B. pendula individual. Gene duplicates from the paleohexaploid event were enriched for transcriptional regulation, whereas tandem duplicates were overrepresented by environmental responses. Population resequencing of 80 individuals showed effective population size crashes at major points of climatic upheaval. Selective sweeps were enriched among polyploid duplicates encoding key developmental and physiological triggering functions, suggesting that local adaptation has tuned the timing of and cross-talk between fundamental plant processes. Variation around the tightly-linked light response genes PHYC and FRS10 correlated with latitude and longitude and temperature, and with precipitation for PHYC. Similar associations characterized the growth-promoting cytokinin response regulator ARR1, and the wood development genes KAK and MED5A., published version, peerReviewed

Methyl jasmonate (MeJA) is widely used as a model chemical to study hypersensitive responses to biotic stress impacts in plants. Elevated levels of methyl jasmonate induce jasmonate-dependent defense responses, associated with a decline in primary metabolism and enhancement of secondary metabolism of plants. However, there is no information of how stress resistance of plants, and accordingly the sensitivity to exogenous MeJA can be decreased by endophytic plant growth promoting rhizobacteria (PGPR) harboring ACC (1-aminocyclopropane-1-carboxylate) deaminase. In this study, we estimated stress alleviating potential of endophytic PGPR against MeJA-induced plant perturbations through assessing photosynthetic traits and stress volatile emissions. We used mild (5 mM) to severe (20 mM) MeJA and endophytic plant growth promoting rhizobacteria Burkholderia vietnamiensis CBMB40 and studied how MeJA and B. vietnamiensis treatments influenced temporal changes in photosynthetic characteristics and stress volatile emissions. Separate application of MeJA markedly decreased photosynthetic characteristics and increased lipoxygenase pathway (LOX) volatiles, volatile isoprenoids, saturated aldehydes, lightweight oxygenated compounds (LOC), geranyl-geranyl diphosphate pathway (GGDP) volatiles, and benzenoids. However, MeJA-treated leaves inoculated by endophytic bacteria B. vietnamiensis had substantially increased photosynthetic characteristics and decreased emissions of LOX, volatile isoprenoids and other stress volatiles compared with non-inoculated MeJA treatments, especially at later stages of recovery. In addition, analysis of leaf terpenoid contents demonstrated that several mono- and sesquiterpenes were de novo synthesized upon MeJA and B. vietnamiensis applications. This study demonstrates that foliar application of endophytic bacteria B. vietnamiensis can potentially enhance resistance to biotic stresses and contribute to the maintenance of the integrity of plant metabolic activity.

Understanding within-species variability in terpenoid content and composition is highly relevant for predicting species adaptive potential to biotic stresses, but there is still limited information on terpene variations even for widespread species. We studied the foliage content and composition of terpenoids, foliage structure, and carbon (C) and nitrogen (N) contents in Pinus sylvestris L. in four Estonian sites ranging from dry forest to raised bog. In the bogs, samples were taken along the environmental gradient from drier margins towards wetter central parts. A chiral column was used to gain insight into the variation in terpene composition. We hypothesized that terpene contents increase and the composition becomes more diverse in more strongly N-limited sites (greater C/N ratio) and that terpene signatures cluster together in sub-sites with similar conditions (drier/wetter). Altogether 37 terpenes were quantified across the sites. Extremely large variability of terpene contents, 48-62% for monoterpenes and 61-89% for sesquiterpenes, was observed. According to the amounts of α-pinenes and (+)-3-carene, we distinguished two different 'pine chemotypes'. Contrary to the hypothesis, terpene contents and variability were the greatest in the dry site with the lowest C/N ratio. However, individual terpenoids correlated differently with C or N in different sites, indicating site effects on terpene composition. Moreover, correlations between the terpenoids and C or N depended on the pine chemotype. The sub-sites with different water regime were more strongly clustered together within the site than across the sites. The study demonstrates extensive variations in terpene contents and composition among the populations and over short spatial distances within the populations, suggesting a large among- and within-population adaptive capacity of P. sylvestris.

Terpene synthases are responsible for synthesis of a large number of terpenes in plants using substrates provided by two distinct metabolic pathways, the mevalonate-dependent pathway that is located in cytosol and has been suggested to be responsible for synthesis of sesquiterpenes (C15), and 2-C-methyl-D-erythritol-4-phosphate pathway located in plastids and suggested to be responsible for the synthesis of hemi- (C5), mono- (C10), and diterpenes (C20). Recent advances in characterization of genes and enzymes responsible for substrate and end product biosynthesis as well as efforts in metabolic engineering have demonstrated existence of a number of multi-substrate terpene synthases. This review summarizes the progress in the characterization of such multi-substrate terpene synthases and suggests that the presence of multi-substrate use might have been significantly underestimated. Multi-substrate use could lead to important changes in terpene product profiles upon substrate profile changes under perturbation of metabolism in stressed plants as well as under certain developmental stages. We therefore argue that multi-substrate use can be significant under physiological conditions and can result in complicate modifications in terpene profiles.

Plant-generated volatiles constitute a sensitive signal of stress response, but quantitative relationships between the stress severity and volatile emissions have been demonstrated only for a few stresses. Among important stresses in the field, chilling and frost stress in spring and heat stress mid-season can significantly curb productivity. We studied the effects of cold and heat shock treatments on leaf photosynthesis and the emission of the volatile products of the lipoxygenase pathway (LOX, also called green leaf volatiles) and mono- and sesquiterpene emissions in tomato (Solanum lycopersicum cv. Mato) to gain quantitative insights into temperature stress-elicited volatile emissions. Both cold and heat stress treatments ranged from mild, which only weakly affected foliage photosynthesis, to severe, which almost completely inhibited photosynthesis. Under non-stressed conditions, LOX emissions were close to the detection limit, and terpene emissions were low. Both cold and heat stress led to enhancement of LOX emissions according to a switch-type response with essentially no emissions under mild stress and major emissions under severe stress. The emissions of mono- and sesquiterpenes increased gradually with the severity of stress, but cold stress resulted in higher sesquiterpene emissions at any given monoterpene emission level. We suggest that the quantitative relationships between the stress strength and emissions observed in this study provide an important means to characterize the severity of cold and heat stresses.

Knowledge of pistachio genetic diversity is necessary for the formulation of appropriate management strategies for the conservation of these species. We analysed amplified fragment length polymorphisms in a total of 216 pistachio accessions, which included seven populations from three wild species (Pistacia vera, Pistacia khinjuk and Pistacia atlantica subsp. kurdica) and most of the important cultivars from Iran, together with some foreign cultivars. High levels of genetic diversity were detected within the Iranian cultivars, and they showed a clear separation from foreign cultivars, as revealed by unweighted pair group method with arithmetic averaging and supported by analysis of molecular variance. The lowest amount of polymorphism was observed in P. atlantica subsp. kurdica, which showed the lowest number of total bands as compared to the other species. This revealed strong genetic erosion of P. atlantica subsp. kurdica, which reflected a severe decline in habitat and over-exploitation. Based on these findings, strategies are proposed for the genetic conservation and management of pistachio species and cultivars.

Iran is one of the two major centres of Pistacia diversity and the main producer of pistachios in the world. About 282 Iranian pistachio genotypes (Pistacia spp.), together with 22 foreign cultivars (P. vera), were genotyped using 10 simple sequence repeat (SSR) markers to analyse the genetic diversity and relationships among Pistacia species and cultivars. The results revealed that the genetic diversity within P. atlantica subsp. kurdica was considerably lower than in P. vera or P. khinjuk. Principal coordinate analysis revealed a clear separation between the different Pistacia spices, as well as between the Iranian and foreign cultivars. AMOVA analysis showed that the variation between the species, between different populations, and within populations accounted for 41, 9, and 50% of the total variation, respectively. The results demonstrated that the study of genetic diversity and relationships among Pistacia species and cultivars using SSR markers provides important information for the collection and conservation of pistachio germplasm. In addition, the Iranian cultivars had a broader genetic background than that of the foreign cultivars. Thus, they are very important for genetic conservation and the planning of future breeding programmes. We also determined the different levels of genetic diversity that exist between and within the species and populations and showed that gene flow occurs between the Iranian cultivars and wild-type P. vera populations. The study provides practical information that policy-makers and scientists can apply to the conservation and sustainable use of all the species studied. © Springer Science+Business Media B.V. 2009.

Fusarium head blight (FHB or head scab) has become a major limiting factor for sustainable wheat (Triticum aestivum L.) production around the world. For quantitative trait loci (QTL) analysis of resistance to FHB, F 3 plants and F 3: 5 lines, derived from a 'Frontana' (moderately resistant)/'Seri82' (susceptible) cross, were spray-inoculated in 2001 and 2002, respectively. Artificial inoculations were carried out under field conditions. Of 273 SSR and AFLP markers, 250 could be mapped and they yielded 42 linkage groups, covering a genetic distance of 1931 cM. QTL analysis was based on the constructed linkage map and area under the disease progress curve (AUDPC). The analyses revealed three consistent QTLs associated with FHB resistance on chromosomes 1 BL, 3AL and 7AS explaining 7.9%, 7.7% and 7.6% of the phenotypic variation, respectively, above 2 years. The results confirmed the previously described resistance QTL of 'Frontana' on chromosome 3AL. A combination of 'Frontana' resistance with 'Sumai-3' resistance may lead to lines with augmented resistance expression.

All tree species possess genes encoding terminal enzymes responsible for volatile isoprenoid synthesis. However, only in some species, these genes are expressed constitutively in leaves, while terpenoid emissions can be triggered by abiotic and biotic stress factor in essentially all species. This chapter analyses the biochemical diversity of volatile isoprenoid synthases and investigates the genomic modifications responsible for constitutive volatile production in trees. Plant terpenoids are up to three-domain proteins with either one active center in monofunctional synthases, or two active centers in bifunctional synthases. There is evidence of monophyletic origin of modern plant terpenoid synthases from a three-domain synthase in an ancient progenitor followed by extensive gene duplication and domain loss. The terpenoid synthase sequence similarity can be low among distant plant groups, but terpenoid tertiary structure is remarkably similar in different synthases, and this structural similarity is even conserved across domains of life. However, only minor changes in active center structure can lead to major changes in product profiles, indicating that presence of rich terpenoid genetic diversity constitutes an important means for rapid evolutionary adaptations to novel biotic interactions, and to new abiotic stresses in plant habitats.