Energetic costs of tissue construction were compared in two subspecies of Phragmites australis, the common reed – namely the primary native and introduced lineages in North America. Caplan et al. report that the introduced lineage has lower construction costs than the native under all environmental conditions assessed, driven mainly by its lower cost rhizomes. These results highlight the fact that belowground energetics, which are seldom investigated, can influence the performance advantages that drive many plant invasions. The authors also demonstrate that tissue construction costs in organs not typically assessed can shift with global change, suggesting that they may have increasingly important implications into the future.
The energetic cost of plant organ construction is a functional trait that is useful for understanding carbon investment during growth (e.g. the resource acquisition vs. tissue longevity tradeoff), as well as in response to global change factors like elevated CO2 and N. Despite the enormous importance of roots and rhizomes in acquiring soil resources and responding to global change, construction costs have been studied almost exclusively in leaves. We sought to determine how construction costs of aboveground and belowground organs differed between native and introduced lineages of a geographically widely dispersed wetland plant species (Phragmites australis) under varying levels of CO2 and N. We grew plants under ambient and elevated atmospheric CO2, as well as under two levels of soil nitrogen. We determined construction costs for leaves, stems, rhizomes and roots, as well as for whole plants. Across all treatment conditions, the introduced lineage of Phragmites had a 4.3 % lower mean rhizome construction cost than the native. Whole-plant construction costs were also smaller for the introduced lineage, with the largest difference in sample means (3.3 %) occurring under ambient conditions. In having lower rhizome and plant-scale construction costs, the introduced lineage can recoup its investment in tissue construction more quickly, enabling it to generate additional biomass with the same energetic investment. Our results suggest that introduced Phragmites has had an advantageous tissue investment strategy under historic CO2 and N levels, which has facilitated key rhizome processes, such as clonal spread. We recommend that construction costs for multiple organ types be included in future studies of plant carbon economy, especially those investigating global change.
Carbon dioxide; common reed; construction cost; eutrophication; intraspecific; invasion ecology; Phragmites; plant functional traits; rhizomes; wetlands.
Gametes have a unique potential to enter the sporophytic pathway, called androgenesis. The plants produced are usually haploid and recombinant due to the preceding meiosis and they can double their chromosome number to form doubled haploids (DHs), which are completely homozygous. The present study demonstrates that reprogramming occurs in the isolated microspores at tetrad stage as the first step if androgenesis and this is paving the way for the development of an efficient technique for the production of homozygous lines in cassava. This is the first ever detailed report of microspore reprogramming at the tetrad stage and the first report of microspore embryogenesis induction in cassava with detailed evidence.
Gametes have the unique potential to enter the sporophytic pathway, called androgenesis. The plants produced are usually haploid and recombinant due to the preceding meiosis and they can double their chromosome number to form doubled haploids, which are completely homozygous. Availability of the doubled haploids facilitates mapping the genes of agronomically important traits, shortening the time of the breeding process required to produce new hybrids and homozygous varieties, and saving the time and cost for inbreeding. This study aimed to test the feasibility of using isolated and in vitro cultured immature cassava (Manihot esculenta) microspores to reprogramme and initiate sporophytic development. Different culture media and different concentrations of two ion components (Cu2+ and Fe2+) were tested in two genotypes of cassava. External structural changes, nuclear divisions and cellular changes during reprogramming were analysed by scanning electron microscopy, by staining with 4′,6-diamidino-2-phenylindole, and through classical histology and transmission electron microscopy. In two cassava genotypes, different developmental stages of microspores were found to initiate sporophytic cell divisions, that is, with tetrads of TMS 60444 and with mid or late uni-nucleate microspores of SM 1219-9. In the modified NLN medium (NLNS), microspore enlargements were observed. The medium supplemented with either sodium ferrous ethylene-diamine-tetraacetic acid (NaFeEDTA) or CuSO4·5H2O induced sporophytic cell division in both genotypes. A low frequency of the reprogramming and the presence of non-responsive microspores among the responsive ones in tetrads were found to be related to the viability and exine formation of the microspores. The present study clearly demonstrated that reprogramming occurs much faster in isolated microspore culture than in anther culture. This paves the way for the development of an efficient technique for the production of homozygous lines in cassava. This is the first ever detailed report of microspore reprogramming at the tetrad stage and the first report of microspore embryogenesis induction in cassava with detailed evidence.
Androgenesis; DAPI; histology; Manihot esculenta; reprogramming; scanning electron microscopy; transmission electron microscopy.
Clonal plants are common in frequently flooded habitats because of their resilience to disturbance. We investigated if submergence prior to fragmentation of clones of two clover species reduced survival and regrowth of clonal fragments, and if these fitness parameters were different between genotypes from highly disturbed river forelands and less disturbed coastal dune slacks. We found that soil flooding severely decreased survival and regrowth, and that plants from the more disturbance-prone habitat were less negatively affected by fragmentation. However, internode size was, surprisingly, often negatively correlated with survival after fragmentation, but positively correlated with regrowth. Apparently, contrasting selection pressures exist on internode size in stoloniferous species growing in disturbed habitats.
Clonal plants, which reproduce by means of stolons and rhizomes, are common in frequently flooded habitats. Resilience to disturbance is an important trait enabling plants to survive in such highly disturbed habitats. Resource storage is thought to enable clonal plants to resume growth after clonal fragmentation caused by disturbance. Here we investigated if submergence prior to disturbance reduces survival and regrowth of clonal fragments and whether or not genotypes originating from highly disturbed riverine habitats are more resistant to mechanical disturbance than genotypes from less disturbed coastal dune slack habitats. We further tested if variation in survival and regrowth was affected by internode size. Clones from contrasting habitats of two closely related Trifolium species were first genotypically characterized by amplification fragment length polymorphism and then subjected to soil flooding and subsequent clonal fragmentation. These species differ with respect to their abundance in riverine and dune slack habitats, with Trifolium repens mainly occurring in riverine grasslands and Trifolium fragiferum in coastal dune slacks. Soil flooding decreased survival and regrowth by up to 80 %. Plants originating from riverine grasslands were less negatively affected by fragmentation than plants from dune slack habitats. Surprisingly, ramets did not always benefit from being attached to a larger internode, as internode size was often negatively correlated with survival after fragmentation. Regrowth, on the other hand, was generally positively correlated with internode size. These unexpected results indicate that there may be contrasting selection pressures on internode size in stoloniferous species growing in severely disturbed habitats.
Clonal growth; disturbance; fragmentation; soil flooding; Trifolium fragiferum; Trifolium repens.
The tonoplast Na+/H+-antiporter and the tonoplast H+-pumps are essential components of salt tolerance in plants. We investigated the transport activity of the Na+/H+-antiporter and the H+-pumps in a highly tolerant salt accumulating halophyte, Salicornia dolichostachya, and compared them with activities in the related glycophyte Spinacia oleracea. Our results suggest that S. dolichostachya generates a high tonoplast H+-gradient already at low external salinities. At high external salinities, S. dolichostachya showed improved growth compared to S. oleracea, but H+-pump and Na+/H+-exchange activities were comparable between the species, which might imply that S. dolichostachya more efficiently retains Na+ in the vacuole.
The tonoplast Na+/H+ antiporter and tonoplast H+ pumps are essential components of salt tolerance in plants. The objective of this study was to investigate the transport activity of the tonoplast Na+/H+ antiporter and the tonoplast V-H+-ATPase and V-H+-PPase in a highly tolerant salt-accumulating halophyte, Salicornia dolichostachya, and to compare these transport activities with activities in the related glycophyte Spinacia oleracea. Vacuolar membrane vesicles were isolated by density gradient centrifugation, and the proton transport and hydrolytic activity of both H+ pumps were studied. Furthermore, the Na+/H+-exchange capacity of the vesicles was investigated by 9-amino-6-chloro-2-methoxyacridine fluorescence. Salt treatment induced V-H+-ATPase and V-H+-PPase activity in vesicles derived from S. oleracea, whereas V-H+-ATPase and V-H+-PPase activity in S. dolichostachya was not affected by salt treatment. Na+/H+-exchange capacity followed the same pattern, i.e. induced in response to salt treatment (0 and 200 mM NaCl) in S. oleracea and not influenced by salt treatment (10 and 200 mM NaCl) in S. dolichostachya. Our results suggest that S. dolichostachya already generates a high tonoplast H+ gradient at low external salinities, which is likely to contribute to the high cellular salt accumulation of this species at low external salinities. At high external salinities, S. dolichostachya showed improved growth compared with S. oleracea, but V-H+-ATPase, V-H+-PPase and Na+/H+-exchange activities were comparable between the species, which might imply that S. dolichostachya more efficiently retains Na+ in the vacuole.
Halophyte; membrane transport; NHX1; Salicornia; salt tolerance; vacuole; V-H+-ATPase; V-H+-PPase.
Genome doubling and changes in genome size are fundamental evolutionary processes. Arabidopsis kamchatica has been reported to contain both diploid and tetraploid individuals (2 or 4 copies of every chromosome). We did find genome size differences among populations, and among populations genome size varied 7%. However, all sampled A. kamchatica plants from a wide geographic range were tetraploids. This level of intraspecific genome size variation in A. kamchatica is lower than in other Arabidopsis taxa. Due to its close relationship to A. thaliana, A. kamchatica has the potential to be very useful in the study of polyploidy and genome evolution.
Polyploidization and subsequent changes in genome size are fundamental processes in evolution and diversification. Little is currently known about the extent of genome size variation within taxa and the evolutionary forces acting on this variation. Arabidopsis kamchatica has been reported to contain both diploid and tetraploid individuals. The aim of this study was to determine the genome size of A. kamchatica, whether there is variation in ploidy and/or genome size in A. kamchatica and to study how genome size has evolved. We used propidium iodide flow cytometry to measure 2C DNA content of 73 plants from 25 geographically diverse populations of the putative allotetraploid A. kamchatica and its parents, Arabidopsis lyrata and Arabidopsis halleri. All A. kamchatica plants appear to be tetraploids. The mean 2C DNA content of A. kamchatica was 1.034 pg (1011 Mbp), which is slightly smaller than the sum of its diploid parents (A. lyrata: 0.502 pg; A. halleri: 0.571 pg). Arabidopsis kamchatica appears to have lost ∼37.594 Mbp (3.6 %) of DNA from its 2C genome. Tetraploid A. lyrata from Germany and Austria appears to have lost ∼70.366 Mbp (7.2 %) of DNA from the 2C genome, possibly due to hybridization with A. arenosa, which has a smaller genome than A. lyrata. We did find genome size differences among A. kamchatica populations, which varied up to 7 %. Arabidopsis kamchatica ssp. kawasakiana from Japan appears to have a slightly larger genome than A. kamchatica ssp. kamchatica from North America, perhaps due to multiple allopolyploid origins or hybridization with A. halleri. However, the among-population coefficient of variation in 2C DNA content is lower in A. kamchatica than in other Arabidopsis taxa. Due to its close relationship to A. thaliana, A. kamchatica has the potential to be very useful in the study of polyploidy and genome evolution.
Allotetraploid; Arabidopsis halleri ssp. gemmifera; Arabidopsis kamchatica; Arabidopsis lyrata; C-value; 2C DNA content; flow cytometry; genome size; genome size variation
Rice is a unique crop plant since it can survive under anoxia condition by flooding and also germinate and grow up to coleoptile through its distinctive adaptations. Despite several decades of research on this topic, the current knowledge on the molecular machinery of rice under anoxia is very limited. Therefore, we unraveled the possible regulatory mechanisms by resorting to systems biology approach which combines the metabolic modeling and transcriptome analysis. Such integrative analysis highlight the critical role of MYB, bZIP, ERF and ZnF transcription factors in up-regulating the fermentation and sucrose metabolism genes to generate sufficient energy for cellular growth.
The ability of rice to germinate under anoxia by extending the coleoptile is a highly unusual characteristic and a key feature underpinning the ability of rice seeds to establish in such a stressful environment. The process has been a focal point for research for many years. However, the molecular mechanisms underlying the anoxic growth of the coleoptile still remain largely unknown. To unravel the key regulatory mechanisms of rice germination under anoxic stress, we combined in silico modelling with gene expression data analysis. Our initial modelling analysis via random flux sampling revealed numerous changes in rice primary metabolism in the absence of oxygen. In particular, several reactions associated with sucrose metabolism and fermentation showed a significant increase in flux levels, whereas reaction fluxes across oxidative phosphorylation, the tricarboxylic acid cycle and the pentose phosphate pathway were down-regulated. The subsequent comparative analysis of the differences in calculated fluxes with previously published gene expression data under air and anoxia identified at least 37 reactions from rice central metabolism that are transcriptionally regulated. Additionally, cis-regulatory content analyses of these transcriptionally controlled enzymes indicate a regulatory role for transcription factors such as MYB, bZIP, ERF and ZnF in transcriptional control of genes that are up-regulated during rice germination and coleoptile elongation under anoxia.
Anoxia; cis-elements; flux sampling; rice; systems biology; transcription factors; transcriptional regulation.
Promotional Statement: White-tailed deer browsing has been implicated in the loss of species diversity from forests throughout eastern North America. We build on this previous research by examining how browsing also affects phylogenetic community structure. With this approach, we can better understand the role of deer browsing in the community assembly process. In browsed plots, we found that reductions in phylogenetic diversity were much greater than reductions in species richness or diversity. Species persisting in browsed communities were also closely-related. Our findings indicate deer browsing acts as a biotic filter during the community assembly process.
Community assembly entails a filtering process, where species found in a local community are those that can pass through environmental (abiotic) and biotic filters and successfully compete. Previous research has demonstrated the ability of white-tailed deer (Odocoileus virginianus) to reduce species diversity and favour browse-tolerant plant communities. In this study, we expand on our previous work by investigating deer as a possible biotic filter altering local plant community assembly. We used replicated 23-year-old deer exclosures to experimentally assess the effects of deer on species diversity (H′), richness (SR), phylogenetic community structure and phylogenetic diversity in paired browsed (control) and unbrowsed (exclosed) plots. Additionally, we developed a deer-browsing susceptibility index (DBSI) to assess the vulnerability of local species to deer. Deer browsing caused a 12 % reduction in H′ and 17 % reduction in SR, consistent with previous studies. Furthermore, browsing reduced phylogenetic diversity by 63 %, causing significant phylogenetic clustering. Overall, graminoids were the least vulnerable to deer browsing based on DBSI calculations. These findings demonstrate that deer are a significant driver of plant community assembly due to their role as a selective browser, or more generally, as a biotic filter. This study highlights the importance of knowledge about the plant tree of life in assessing the effects of biotic filters on plant communities. Application of such knowledge has considerable potential to advance our understanding of plant community assembly.
Browsing; herbivory; phylogenetic clustering; phylogenetic community ecology; plant–animal interactions; species diversity.
Gibberellin (GA) is one of the plant hormones which regulates many aspects of plant growth and developmental processes. Rice plants known as deepwater rice can survive during flooding by elongating its internodes to avoid anoxia. Previous studies reported that GA is essential for internode elongation in deepwater rice. However, the interaction between internode elongation and regulator of GA sensitivity is unknown. In this study, we performed a QTL analysis and identified the chromosomal regions that regulate GA responsiveness in deepwater rice. We concluded that deepwater rice could induce internode elongation in response to GA by factors in these regions.
Gibberellin (GA) is a plant hormone that has important roles in numerous plant developmental phases. Rice plants known as deepwater rice respond to flooding by elongating their internodes to avoid anoxia. Previous studies reported that GA is essential for internode elongation in deepwater rice. Quantitative trait locus (QTL) analyses identified QTLs regulating internode elongation in response to deepwater conditions. However, the interaction between internode elongation and regulators of GA sensitivity in deepwater rice is unknown. In this study, we applied GA to recombinant inbred lines of T65 (non-deepwater rice) and Bhadua (deepwater rice), and performed a QTL analysis of internode elongation in response to GA. GA-induced internode elongation was detected only in deepwater rice. Our QTL analysis revealed two major QTLs on chromosomes 3 and 9 regulating total internode length, lowest elongated internode and number of elongated internodes. Furthermore, the QTL on chromosome 3 acted as an enhancer of other QTLs (e.g. the QTL on chromosome 12). Nearly isogenic lines of deepwater rice carrying the QTL regions from chromosomes 3 and 12 of the deepwater rice C9285 showed internode elongation in response to GA. Thus, these QTLs may regulate GA responsiveness in deepwater rice. This study furthers our understanding of the mechanism of internode elongation in rice.
Deepwater rice; gibberellin response; internode elongation; QTL.
Taxa from the genus Prosopis are widespread invasive aliens across the globe. Numerous species have contentious issues surrounding them as they provide both benefits and harm. Prosopis taxa are currently naturalised or invasive in 103 countries and are bioclimatically suitable for many more. There are numerous management practices available to control Prosopis invasions, each with their benefits and costs, however, in most areas management has had only limited success. There is need for more research to improve understanding and management success and for countries to develop strategic plants to guide managed in the future.
Invasive species cause ecological, economic and social impacts and are key drivers of global change. This is the case for the genus Prosopis (mesquite; Fabaceae) where several taxa are among the world's most damaging invasive species. Many contentious issues (‘conflicts of interest’) surround these taxa, and management interventions have not yet sustainably reduced the negative impacts. There is an urgent need to better understand the factors that drive invasions and shape management actions, and to compare the effectiveness of different management approaches. This paper presents a global review of Prosopis, focusing on its distribution, impacts, benefits and approaches to management. Prosopis was found to occur in a 129 countries globally and many more countries are climatically suitable. All areas with naturalized or invasive Prosopis species at present are suitable for more taxa and many Asian and Mediterranean countries with no records of Prosopis are bioclimatically suitable. Several Prosopis species have substantial impacts on biodiversity, ecosystem services, and local and regional economies in their native and even more so in their invasive ranges; others provide multiple benefits to local communities. Management efforts are underway in only a small part of the invaded range. Countries where more research has been done are more likely to implement formal management than those where little published research is available. Management strategies differ among countries; developed nations use mainly mechanical and chemical control whereas developing nations tend to apply control through utilization approaches. A range of countries are also using biological control. Key gaps in knowledge and promising options for management are highlighted.
Classification and regression tree; distribution; global review; impacts; logistic regression; management; mesquite; tree invasions.
This research examines the contribution of plant height, number of flowers, number of stems, as well the joint impacts of mutualists and antagonists on the pollination biology and seed production of the imperiled, deceptive orchid, Cypripedium candidum. We found flowering stem height to be the only morphological feature significant in reproduction, with taller flowering stems simultaneously receiving increased pollination and decreased seed predation. Furthermore we found decreased seed mass in individuals subjected to hand-self pollination treatments. Our results may help explain the factors limiting seed production in other Cypripedium and further emphasize the importance of management in orchid conservation.
For many species of conservation significance, multiple factors limit reproduction. This research examines the contributions of plant height, number of flowers, number of stems, pollen limitation and seed predation to female reproductive success in the deceit-pollinated orchid, Cypripedium candidum. The deceptive pollination strategy employed by many orchids often results in high levels of pollen limitation. While increased floral display size may attract pollinators, C. candidum's multiple, synchronously flowering stems could promote selfing and also increase attack by weevil seed predators. To understand the joint impacts of mutualists and antagonists, we examined pollen limitation, seed predation and the effects of pollen source over two flowering seasons (2009 and 2011) in Ohio. In 2009, 36 pairs of plants size-matched by flower number, receiving either supplemental hand or open pollination, were scored for fruit maturation, mass of seeds and seed predation. Pollen supplementation increased proportion of flowers maturing into fruit, with 87 % fruit set when hand pollinated compared with 46 % for naturally pollinated flowers. Inflorescence height had a strong effect, as taller inflorescences had higher initial fruit set, while shorter stems had higher predation. Seed predation was seen in 73 % of all fruits. A parallel 2011 experiment that included a self-pollination treatment and excluded seed predators found initial and final fruit set were higher in the self and outcross pollination treatments than in the open-pollinated treatment. However, seed mass was higher in both open pollinated and outcross pollination treatments compared with hand self-pollinated. We found greater female reproductive success for taller flowering stems that simultaneously benefited from increased pollination and reduced seed predation. These studies suggest that this species is under strong reinforcing selection to increase allocation to flowering stem height. Our results may help explain the factors limiting seed production in other Cypripedium and further emphasize the importance of management in orchid conservation.
Conservation; orchid; plant reproduction; plant–insect interactions; pollen limitation; pollination ecology; reproductive trade-offs; seed predation; supplemental pollination.
Coastal cliff-tops are specific saline environments, where only highly specialized halophytes can thrive. Limonium spp. are commonly found in these ecological conditions, many of them being considered as threatened or with an unknown conservation status. The habitat requirements of Limonium multiflorum, an apomictic halophyte endemic to western Portugal, were investigated. Results showed the species narrow habitat specificity as well as its intolerance to competition with invasive alien plants. We conclude that in situ conservation of this rare and vulnerable species emerges as a priority in order to ensure that its biodiversity is not lost.
Coastal areas and other saline environments are major contributors to regional and global biodiversity patterns. In these environments, rapidly changing gradients require highly specialized plants like halophytes. In European coastal cliff-tops, rocky and sandy seashores, and saltmarshes, typical halophytes from the genus Limonium are commonly found. Among them, the aneuploid tetraploid (2n = 4x = 35, 36, 37) Limonium multiflorum, endemic to the west coast of Portugal, is an interesting case study for investigating the ecology and conservation of a halophyte agamospermic species. Although it is listed in the IUCN red list of threatened species, information on its population size or rarity, as well as its ecology, in some respects is still unknown. Field surveys in the largest known population were performed (Raso cape, Portugal) in order to determine habitat requirements and conservation status. A total of 88 quadrats were monitored, 43 of which contained at least one L. multiflorum individual. For each sampled quadrat, four abiotic and four biotic variables as well as two spatially derived variables were recorded. Principal component analysis and cluster analysis showed narrow habitat specificity for this species which appeared to be intolerant to competition with invasive alien plants. We conclude that in situ conservation in a local ‘hotspot’ of this rare and vulnerable species emerges as a priority in order to ensure that biodiversity is not lost.
Agamospermic species; cliff-dwelling species; conservation; habitat specificity; halophyte; Limonium.
Salt marsh grasses are adapted to thrive under saline conditions by various combinations of traits. Some researchers suggested that salt excreting grasses would differ from non-excreting ones in these traits. However, little is known about the differential responses between these plant types. Here, we compared the growth and physiology of salt excreting and non-excreting grasses. Differences were found between the two grass types in leaf water content, accumulation of organic compounds and Na+ distribution which appeared to be linked with salt excretion. Additional studies on a number of halophytic grasses could help to identify key traits for salt resistance.
The combination of traits that makes a plant successful under saline conditions varies with the type of plant and its interaction with the environmental conditions. Knowledge about the contribution of these traits towards salt resistance in grasses has great potential for improving the salt resistance of conventional crops. We attempted to identify differential adaptive response patterns of salt-excreting versus non-excreting grasses. More specifically, we studied the growth, osmotic, ionic and nutrient (carbon/nitrogen) relations of two salt-excreting (Aeluropus lagopoides and Sporobolus tremulus) and two non-excreting (Paspalum paspalodes and Paspalidium geminatum) perennial C4 grasses under non-saline and saline (0, 200 and 400 mM NaCl) conditions. Growth and relative growth rate decreased under saline conditions in the order P. geminatum > S. tremulus = A. lagopoides > P. paspalodes. The root-to-shoot biomass allocation was unaffected in salt-excreting grasses, increased in P. paspalodes but decreased in P. geminatum. Salt-excreting grasses had a higher shoot/root Na+ ratio than non-excreting grasses. K+, Ca2+ and Mg2+ homoeostasis remained undisturbed among test grasses possibly through improved ion selectivity with rising substrate salinity. Salt-excreting grasses increased leaf succulence, decreased ψs and xylem pressure potential, and accumulated proline and glycinebetaine with increasing salinity. Higher salt resistance of P. paspalodes could be attributed to lower Na+ uptake, higher nitrogen-use efficiency and higher water-use efficiency among the test species. However, P. geminatum was unable to cope with salt-induced physiological drought. More information is required to adequately document the differential strategies of salt resistance in salt-excreting and non-excreting grasses.
C–N balance; compatible solutes; halophytic grasses; ion homoeostasis; Na+ flux; nitrogen-use efficiency.
One may have seen wintering rosette leaves totally frozen and wilted in the early morning but recover during the daytime. How can cold hardy plants survive freezing of the tissues, unlike animal tissues? Cold hardy plants seem to have evolved various strategies. One example is extracellular freezing, where icicles primarily form in intercellular spaces whilst the cells are dehydrated, yet the underlying mechanisms remain unclear. In this study, using blueberry stems, we found high ice nucleation activity specifically localized in the cell wall fraction of bark tissues. This activity likely contributes to the primary and spontaneous initiation of freezing in the intercellular spaces of the bark to successfully perform extracellular freezing.
Controlled ice nucleation is an important mechanism in cold-hardy plant tissues for avoiding excessive supercooling of the protoplasm, for inducing extracellular freezing and/or for accommodating ice crystals in specific tissues. To understand its nature, it is necessary to characterize the ice nucleation activity (INA), defined as the ability of a tissue to induce heterogeneous ice nucleation. Few studies have addressed the precise localization of INA in wintering plant tissues in respect of its function. For this purpose, we recently revised a test tube INA assay and examined INA in various tissues of over 600 species. Extremely high levels of INA (−1 to −4 °C) in two wintering blueberry cultivars of contrasting freezing tolerance were found. Their INA was much greater than in other cold-hardy species and was found to be evenly distributed along the stems of the current year's growth. Concentrations of active ice nuclei in the stem were estimated from quantitative analyses. Stem INA was localized mainly in the bark while the xylem and pith had much lower INA. Bark INA was located mostly in the cell wall fraction (cell walls and intercellular structural components). Intracellular fractions had much less INA. Some cultivar differences were identified. The results corresponded closely with the intrinsic freezing behaviour (extracellular freezing) of the bark, icicle accumulation in the bark and initial ice nucleation in the stem under dry surface conditions. Stem INA was resistant to various antimicrobial treatments. These properties and specific localization imply that high INA in blueberry stems is of intrinsic origin and contributes to the spontaneous initiation of freezing in extracellular spaces of the bark by acting as a subfreezing temperature sensor.
Blueberry; cold hardiness; extracellular freezing; freezing tolerance; ice nucleation; infra-red thermography; Vaccinium ashei; Vaccinium corymbosum
Fruit quality and more specifically quality of the fiber in the fruit of cotton, depends on interactions between fruit position in the plant architecture, temperature and agronomical practices, such as sowing time, mulching with plastic film, and topping of the plant's main stem and branches. A functional and structural cotton model CottonXL for fiber quality (strength, length and micronaire) was implemented at the level of each individual fruit in relation to thermal time for optimizing cotton fiber quality by matching cotton management to the environment. The model may be used to address climate and land use change scenarios.
In general, the quality of fruits depends on local conditions experienced by the fruit during its development. In cotton, fruit quality, and more specifically the quality of the fibre in the fruit, depends on interactions between fruit position in the plant architecture, temperature and agronomic practices, such as sowing time, mulching with plastic film and topping of the plant's main stem and branches. To quantify this response of cotton fibre quality to environment and management, we developed a simulation model of cotton growth and development, CottonXL. Simulation of cotton fibre quality (strength, length and micronaire) was implemented at the level of each individual fruit, in relation to thermal time (represented by physiological age of the fruit) and prevailing temperature during development of each fruit. Field experiments were conducted in China in 2007 to determine model parameters, and independent data on cotton fibre quality in three cotton producing regions in China were used for model validation. Simulated values for fibre quality closely corresponded to experimental data. Scenario studies simulating a range of management practices predicted that delaying topping times can significantly decrease fibre quality, while sowing date and film mulching had no significant effect. We conclude that CottonXL may be used to explore options for optimizing cotton fibre quality by matching cotton management to the environment, taking into account responses at the level of individual fruits. The model may be used at plant, crop and regional levels to address climate and land-use change scenarios.
Cotton (Gossypium hirsutum); fibre length; fibre strength; functional–structural plant model (FSPM); growth and development; micronaire; simulation model.
In this study, we determined whether nickel uptake varies with level of serpentine endemism and quantified nickel concentrations in leaves, pistils, anthers, pollen and nectar in several plant species. Endemic species had the lowest tissue concentrations of nickel. Species indifferent to serpentines incorporated higher concentrations of nickel into reproductive organs relative to leaves, but this was not the case for indicator species and endemics where nickel was similar in these organs. Our findings suggest that endemic species possess the ability to limit nickel uptake into above-ground tissues, particularly in reproductive organs where it may interfere with survival and reproduction.
Serpentine soils are edaphically stressful environments that host many endemic plant species. In particular, serpentine soils are high in several heavy metals (e.g. nickel, cobalt and chromium) and these high heavy metal concentrations are thought, in part, to lead to varying levels of plant adaptation and soil affinities (i.e. endemic vs. non-endemic plant species). It is unclear, however, whether serpentine endemics vs. non-endemics differ with respect to heavy metal uptake into either vegetative or reproductive organs. Here, we use nickel as a model to determine whether plant heavy metal uptake varies with the level of endemism in several non-hyperaccumulating species. Under controlled greenhouse conditions, we grew seven plant species from the Brassicaceae family that vary in their degrees of affinity to serpentine soil from low (indifferent) to medium (indicator) and high (endemic) in soil that was nickel supplemented or not. We quantified nickel concentrations in leaves, pistils, anthers, pollen and nectar. While nickel concentrations did not vary across organs or affinities when grown in control soils, under conditions of nickel supplementation endemic species had the lowest tissue concentrations of nickel, particularly when considering leaves and pistils, compared with indifferent/indicator species. Species indifferent to serpentines incorporated higher concentrations of nickel into reproductive organs relative to leaves, but this was not the case for indicator species and endemics where nickel concentration was similar in these organs. Our findings suggest that endemic species possess the ability to limit nickel uptake into above-ground tissues, particularly in reproductive organs where it may interfere with survival and reproduction. Indifferent species accumulated significantly more nickel into reproductive organs compared with leaves, which may limit their reproductive potential relative to endemic species when growing on serpentine soils. Additional work determining the fitness consequences of these differences will further our understanding of edaphic endemism.
Brassicaceae; endemism; flowers; metal accumulation; nickel; serpentine soil.
Banana is the fourth most important food commodity of the world and forms the staple food of majority of people in the tropical and subtropical regions. Banana production is severely constrained by Fusarium wilt disease that causes enormous loss. The present study developed transgenic banana plants overexpressing native cell death genes to impart Fusarium wilt resistance. Since banana is predominantly a vegetatively propagated crop, genetic engineering is the most viable option for development of resistance against important diseases of this crop.
In order to feed an ever-increasing world population, there is an urgent need to improve the production of staple food and fruit crops. The productivity of important food and fruit crops is constrained by numerous biotic and abiotic factors. The cultivation of banana, which is an important fruit crop, is severely threatened by Fusarium wilt disease caused by infestation by an ascomycetes fungus Fusarium oxysporum f. sp. cubense (Foc). Since there are no established edible cultivars of banana resistant to all the pathogenic races of Foc, genetic engineering is the only option for the generation of resistant cultivars. Since Foc is a hemibiotrophic fungus, investigations into the roles played by different cell-death-related genes in the progression of Foc infection on host banana plants are important. Towards this goal, three such genes namely MusaDAD1, MusaBAG1 and MusaBI1 were identified in banana. The study of their expression pattern in banana cells in response to Foc inoculation (using Foc cultures or fungal toxins like fusaric acid and beauvericin) indicated that they were indeed differentially regulated by fungal inoculation. Among the three genes studied, MusaBAG1 showed the highest up-regulation upon Foc inoculation. Further, in order to characterize these genes in the context of Foc infection in banana, we generated transgenic banana plants constitutively overexpressing the three genes that were later subjected to Foc bioassays in a contained greenhouse. Among the three groups of transgenics tested, transformed banana plants overexpressing MusaBAG1 demonstrated the best resistance towards Foc infection. Further, these plants also showed the highest relative overexpression of the transgene (MusaBAG1) among the three groups of transformed plants generated. Our study showed for the first time that native genes like MusaBAG1 can be used to develop transgenic banana plants with efficient resistance towards pathogens like Foc.
Banana; disease resistance; Fusarium wilt; PCD; transgenic; UPR.
Picconia azorica is an endangered endemic species of the Azores whose hard and high density wood is very appreciated for the production of toys, agricultural tools, furniture and religious statuary. Its renewed economic interest represents a good opportunity for establishing conservation programmes. To contribute with information useful for the decision making we performed the genetic analysis of 230 samples from 11 populations collected in three Azorean islands. The majority of the genetic variability was found within populations and no genetic structure was detected between populations and between islands, indicating that the oceanic barriers do not greatly affect gene flow.
Knowledge of the levels and distribution of genetic diversity is important for designing conservation strategies for threatened and endangered species so as to guarantee sustainable survival of populations and to preserve their evolutionary potential. Picconia azorica is a valuable Azorean endemic species recently classified as endangered. To contribute with information useful for the establishment of conservation programmes, the genetic variability and differentiation among 230 samples from 11 populations collected in three Azorean islands was accessed with eight inter-simple sequence repeat markers. A total of 64 polymorphic loci were detected. The majority of genetic variability was found within populations and no genetic structure was detected between populations and between islands. Also the coefficient of genetic differentiation and the level of gene flow indicate that geographical distances do not act as barriers for gene flow. In order to ensure the survival of populations in situ and ex situ management practices should be considered, including artificial propagation through the use of plant tissue culture techniques, not only for the restoration of habitat but also for the sustainable use of its valuable wood.
Azores; endemism conservation; germplasm management; molecular marker; phylogeography; population genetics.
Soil salinization is a serious problem in agricultural lands worldwide. Understanding the mechanisms of salt-tolerant plants will contribute to knowledge necessary to genetically engineer salt-tolerant crops that grow on these saline lands. We identified a genotype of Sporobolus virginicus, a salt-tolerant turf grass, that showed a salinity tolerance to up to a three-fold higher NaCl concentration than seawater salinity. In addition to salt secretion from salt glands on the leaves, this genotype accumulated K+ and proline, a compatible solute, to higher levels than other genotypes under salinity. These properties must contribute to the advanced salt tolerance of this genotype.
Understanding the mechanisms used by halophytic members of the Poaceae to cope with salt stress will contribute to the knowledge necessary to genetically engineer salt-tolerant crops. In this study, we identified a genotype of Sporobolus virginicus, a halophytic turf grass collected in Japan, and investigated its growth rate, ion concentration and secretion, and proline concentration in comparison with the reported properties of genotypes collected from the USA, South Africa and Egypt. Surprisingly, the Japanese genotype showed a salinity tolerance up to 1.5 M NaCl, a 3-fold higher concentration than seawater salinity. Shoot growth was stimulated by 100 mM NaCl and root growth was stimulated at salinities of up to 1 M NaCl. Accumulation of Na+ and CI− in shoots and roots was rapidly elevated by salinity stress but did not exceed the levels required for osmotic adjustment, due in part to ion secretion by salt glands, which are present in genotypes of S. virginicus. However, the Japanese genotypes accumulated K+ to a higher level than other genotypes, resulting in a relatively high K+/Na+ ratio even under salinity stress. An increase in proline concentration was observed that was proportional to the NaCl concentration in the culture solution and might partially account for osmotic adjustment in the shoots. We also generated and characterized cultured cells of S. virginicus. In 500 mM NaCl, the cultured cells showed an enhanced growth compared with cultured cells of rice. The concentration of Na+ and CI− in the cultured cells in 300–500 mM NaCl was lower than in 100 mM NaCl. Cultured cells of S. virginicus accumulated proline to higher levels than rice cells cultured under salinity stress. The active regulation of Na+, Cl− and K+ influx/efflux and proline accumulation might be involved in salt tolerance mechanisms at the cellular level as well as in planta.
Chloride ion; cultured cells; halophyte; potassium ion; proline; salt tolerance; sodium ion; Sporobolus virginicus
Although aerenchyma formed in the roots of some species can be promoted by ethylene, such roots also form a somewhat less extensive aerenchyma under well-aerated conditions. It has been unclear whether ethylene is involved in promoting this constitutive aerenchyma. To test this possibility a novel sandwich method was employed in rice roots. A more extensive aerenchyma was formed on the ACC-treated side. 1-MCP inhibited aerenchyma formation in the presence or absence of ACC. The results indicate that ethylene signaling is involved in aerenchyma development in rice roots and that this may include the regulation of constitutive aerenchyma.
Although the extent of aerenchyma (interconnected gas-filled space) formed in the cortex of the roots of some species can be promoted by submergence and ethylene, such roots also form a somewhat less extensive aerenchyma under well-aerated conditions. It has been unclear whether or not ethylene is also involved in promoting this constitutive aerenchyma. To confirm the potential of ethylene to stimulate aerenchyma development and test the possibility that gas regulates constitutive aerenchyma, a novel sandwich method was employed in rice roots. This involved germinating japonica rice (Oryza sativa) caryopses sandwiched between two agar slabs with or without 1-aminocyclopropane-1-carboxylic acid (ACC) at 1 µM. The roots were then grown for 4 days in the dark in the presence or absence of gaseous 1-methylcyclopropene (1-MCP), an inhibitor of ethylene action. Examination of aerenchyma development by light microscopy demonstrated a more extensive aerenchyma in cross-section on the ACC-treated side that also commenced closer to the root tip. In the presence of 1-MCP at 0.1 or 1 ppm, aerenchyma formation was inhibited in the presence or absence of ACC. 1-Methylcyclopropene also overcame ACC-inhibited root elongation. The results indicate that ethylene signalling is involved in aerenchyma development in primary roots of rice and that this may include the regulation of constitutive aerenchyma. In addition, root elongation was slowed in control roots in the presence of 1 ppm 1-MCP, supporting previous studies demonstrating that endogenous levels of ethylene stimulate root elongation.
Aerenchyma; 1-aminocyclopropane-1-carboxylic acid (ACC); ethylene; 1-methylcyclopropene (1-MCP); Oryza sativa; roots
In several genus evolutionary chromosome change involves variation in DNA amount in diploids and genome downsizing in polyploids. The constancy of bimodal karyotypes, even with changes in ploidy level and DNA content per basic genome indicate that the distribution of DNA within the complement is not at random and suggest the presence of mechanisms selecting for constancy, or against changes, in the karyotype morphology.
Evolutionary chromosome change involves significant variation in DNA amount in diploids and genome downsizing in polyploids. Genome size and karyotype parameters of Hippeastrum species with different ploidy level were analysed. In Hippeastrum, polyploid species show less DNA content per basic genome than diploid species. The rate of variation is lower at higher ploidy levels. All the species have a basic number x = 11 and bimodal karyotypes. The basic karyotypes consist of four short metacentric chromosomes and seven large chromosomes (submetacentric and subtelocentric). The bimodal karyotype is preserved maintaining the relative proportions of members of the haploid chromosome set, even in the presence of genome downsizing. The constancy of the karyotype is maintained because changes in DNA amount are proportional to the length of the whole-chromosome complement and vary independently in the long and short sets of chromosomes. This karyotype constancy in taxa of Hippeastrum with different genome size and ploidy level indicates that the distribution of extra DNA within the complement is not at random and suggests the presence of mechanisms selecting for constancy, or against changes, in karyotype morphology.
Bimodal karyotype; DNA amount variation; genome size; Hippeastrum; karyotype constancy; polyploids.
We studied the halophytic shrub Prosopis strombulifera to investigate whether the differential ability of this species to grow under increasing salt concentrations and mixtures was related to the synthesis of polyphenolic compounds and to the maintenance of leaf pigment contents for an adequate photosynthetic activity. The significant accumulation of flavonoids in tissues under Na2SO4 treatment and their powerful antioxidant activity indicates a role for these compounds in counteracting the oxidative damage induced by severe salt stress, particularly, ionic stress. We demonstrate that ionic interactions between different salts in salinized soils modify the biochemical and morpho-physiological responses of Prosopis strombulifera plants to salinity.
Salt stress conditions lead to increased production of reactive oxygen species (ROS) in plant cells. Halophytes have the ability to reduce these toxic ROS by means of a powerful antioxidant system that includes enzymatic and non-enzymatic components. In this research, we used the halophytic shrub Prosopis strombulifera to investigate whether the ability of this species to grow under increasing salt concentrations and mixtures was related to the synthesis of polyphenolic compounds and to the maintenance of leaf pigment contents for an adequate photosynthetic activity. Seedlings of P. strombulifera were grown hydroponically in Hoagland's solution, gradually adding Na2SO4 and NaCl separately or in mixtures until reaching final osmotic potentials of −1, −1.9 and −2.6 MPa. Control plants were allowed to develop in Hoagland's solution without salt. Oxidative damage in tissues was determined by H2O2 and malondialdehyde content. Leaf pigment analysis was performed by high-performance liquid chromatography with ultraviolet, and total phenols, total flavonoids, total flavan-3-ols, condensed tannins, tartaric acid esters and flavonols were spectrophotometrically assayed. Treatment with Na2SO4 increased H2O2 production and lipid peroxidation in tissues and induced a sharp increase in flavonoid compounds (mainly flavan-3-ols) and consequently in the antioxidant activity. Also, Na2SO4 treatment induced an increased carotenoid/chlorophyll ratio, which may represent a strategy to protect photosystems against photooxidation. NaCl treatment, however, did not affect H2O2 content, lipid peroxidation, pigments or polyphenols synthesis. The significant accumulation of flavonoids in tissues under Na2SO4 treatment and their powerful antioxidant activity indicates a role for these compounds in counteracting the oxidative damage induced by severe salt stress, particularly, ionic stress. We demonstrate that ionic interactions between different salts in salinized soils modify the biochemical and morpho-physiological responses of P. strombulifera plants to salinity.
NaCl; Na2SO4; oxidative damage; pigments; polyphenols; salt stress.
If you add CO2 or nitrogen to a single plant it will likely grow more, but the amount by which each resource stimulates growth differs widely across species. When you add either resource to a whole ecosystem, total plant growth will likely also increase, but there will be winners and losers, causing a change in the relative abundance of plant species, and therefore altering the way the whole ecosystem responds to the added resource, a “community feedback”. These feedbacks are very difficult to predict, especially when multiple resources are added, but a lot of recent experimental evidences suggests that community feedbacks will determine how future ecosystems operate.
While short-term plant responses to global change are driven by physiological mechanisms, which are represented relatively well by models, long-term ecosystem responses to global change may be determined by shifts in plant community structure resulting from other ecological phenomena such as interspecific interactions, which are represented poorly by models. In single-factor scenarios, plant communities often adjust to increase ecosystem response to that factor. For instance, some early global change experiments showed that elevated CO2 favours plants that respond strongly to elevated CO2, generally amplifying the response of ecosystem productivity to elevated CO2, a positive community feedback. However, most ecosystems are subject to multiple drivers of change, which can complicate the community feedback effect in ways that are more difficult to generalize. Recent studies have shown that (i) shifts in plant community structure cannot be reliably predicted from short-term plant physiological response to global change and (ii) that the ecosystem response to multi-factored change is commonly less than the sum of its parts. Here, we survey results from long-term field manipulations to examine the role community shifts may play in explaining these common findings. We use a simple model to examine the potential importance of community shifts in governing ecosystem response. Empirical evidence and the model demonstrate that with multi-factored change, the ecosystem response depends on community feedbacks, and that the magnitude of ecosystem response will depend on the relationship between plant response to one factor and plant response to another factor. Tradeoffs in the ability of plants to respond positively to, or to tolerate, different global change drivers may underlie generalizable patterns of covariance in responses to different drivers of change across plant taxa. Mechanistic understanding of these patterns will help predict the community feedbacks that determine long-term ecosystem responses.
CO2 fertilization; ecological tradeoffs; elevated CO2; multiple factors; nitrogen pollution; plant productivity
We have studied the responses to changing environmental conditions of five halophytes in a Mediterranean salt marsh, during a 2-year period. Salt tolerance in succulent dicotyledonous halophytes is mostly dependent on compartmentalisation of toxic ions in vacuoles and biosynthesis of osmolytes for osmotic adjustment – mechanisms that appear to be constitutive in the most tolerant taxa – while monocots avoid excessive ion transport to the plant aerial parts. Contrary to what has been described for salt treatments under artificial conditions, the selected halophytes are not affected by oxidative stress in their natural habitat, and do not need to activate antioxidant defence mechanisms.
In their natural habitats, different mechanisms may contribute to the tolerance of halophytes to high soil salinity and other abiotic stresses, but their relative contribution and ecological relevance, for a given species, remain largely unknown. We studied the responses to changing environmental conditions of five halophytes (Sarcocornia fruticosa, Inula crithmoides, Plantago crassifolia, Juncus maritimus and J. acutus) in a Mediterranean salt marsh, from summer 2009 to autumn 2010. A principal component analysis was used to correlate soil and climatic data with changes in the plants' contents of chemical markers associated with stress responses: ions, osmolytes, malondialdehyde (MDA, a marker of oxidative stress) and antioxidant systems. Stress tolerance in S. fruticosa, I. crithmoides and P. crassifolia (all succulent dicots) seemed to depend mostly on the transport of ions to aerial parts and the biosynthesis of specific osmolytes, whereas both Juncus species (monocots) were able to avoid accumulation of toxic ions, maintaining relatively high K+/Na+ ratios. For the most salt-tolerant taxa (S. fruticosa and I. crithmoides), seasonal variations of Na+, Cl−, K+ and glycine betaine, their major osmolyte, did not correlate with environmental parameters associated with salt or water stress, suggesting that their tolerance mechanisms are constitutive and relatively independent of external conditions, although they could be mediated by changes in the subcellular compartmentalization of ions and compatible osmolytes. Proline levels were too low in all the species to possibly have any effect on osmotic adjustment. However—except for P. crassifolia—proline may play a role in stress tolerance based on its ‘osmoprotectant’ functions. No correlation was observed between the degree of environmental stress and the levels of MDA or enzymatic and non-enzymatic antioxidants, indicating that the investigated halophytes are not subjected to oxidative stress under natural conditions and do not, therefore, need to activate antioxidant defence mechanisms.
Drought; Inula crithmoides; Juncus acutus; Juncus maritimus; littoral salt marsh; Mediterranean climate; oxidative stress; Plantago crassifolia; Sarcocornia fruticosa; soil salinity.
Plant barcoding uses short DNA sequences to identify unknown samples at species level. This technique relies on the universality of these gene regions and the existence of enough variation among species to allow discrimination. Island radiations pose one challenging scenario where insufficient variation has accumulated in recently diverged groups to allow species identification. In this work we tested whether six gene regions are suitable for barcoding such a radiation in the Macaronesian Lotus. We found high levels of species discrimination in lineages of 3.5 Mya old or older and that the efficiency drastically reduces for younger radiations.
Plant DNA barcoding currently relies on the application of a two-locus combination, matK + rbcL. Despite the universality of these two gene regions across plants, it is suspected that this combination might not have sufficient variation to discriminate closely related species. In this study, we tested the performance of this two-locus plant barcode along with the additional plastid regions trnH-psbA, rpoC1 and rpoB and the nuclear region internal transcribed spacer (nrITS) in a group of 38 species of Lotus from the Macaronesian region. The group has radiated into the five archipelagos within this region from mid-Miocene to early Pleistocene, and thus provides both early divergent and recent radiations that pose a particularly difficult challenge for barcoding. The group also has 10 species considered under different levels of conservation concern. We found different levels of species discrimination depending on the age of the lineages. We obtained 100 % of the species identification from mainland Africa and Cape Verde when all six regions were combined. These lineages radiated >4.5 Mya; however, in the most recent radiations from the end of the Pliocene to the mid-Pleistocene (3.5–1.5 Mya), only 30 % of the species were identified. Of the regions examined, the intergenic region trnH-psbA was the most variable and had the greatest discriminatory power (18 %) of the plastid regions when analysed alone. The nrITS region was the best region when analysed alone with a discriminatory power of 26 % of the species. Overall, we identified 52 % of the species and 30 % of the endangered or threatened species within this group when all six regions were combined. Our results are consistent with those of other studies that indicate that additional approaches to barcoding will be needed in recently evolved groups, such as the inclusion of faster evolving regions from the nuclear genome.
Conservation; DNA barcoding; island radiation; Lotus; Macaronesia; species identification.
We observed that mycorrhizal communities were more divergent among closely related plant species than among distantly related plant species. This was counter to the observation that plant mutualists (e.g. pollinators, seed dispersers) are often shared among closely related host plant species. Since mycorrhizae may affect nutrient competition among neighboring plants, closely related plant neighbors that associate with unique mycorrhizae may have greater functional complementarity and a greater capacity to coexist.
Neighbouring plants are known to vary from having similar to dissimilar arbuscular mycorrhizal fungal (AMF) communities. One possibility is that closely related plants have more similar AMF communities than more distantly related plants, an indication of phylogenetic host specificity. Here, we investigated the structure of AMF communities among dominant grassland plants at three sites in the Northern Great Plains to test whether the pairwise phylogenetic distance among plant species was correlated with pairwise AMF community dissimilarity. For eight dominant and co-occurring grassland plant species, we reconstructed a phylogeny based on DNA data and characterized the AMF communities of their roots at each site. Community analyses revealed that AMF communities varied among sites and among plant species. Contrary to expectations for phylogenetic host specificity, we found that within a site more closely related plants had more distinct AMF communities despite their having similar phenologies. Associations with unique AMF communities may enhance the functional complementarity of related species and promote their coexistence.
Functional complementarity; host identity; mixed-grass prairie; mycorrhizal community structure; niche partitioning; phylogenetic host specificity; phylogenetic signal; plant–soil (below-ground) interactions.