Pedro E. Gundel

Endophytic mediation of reactive oxygen species and antioxidant activity in plants: a review

Reactive oxygen species are in all types of organisms from microbes to higher plants and animals. They are by-products of normal metabolism, such as photosynthesis and respiration, and are responsive to abiotic and biotic stress. Accumulating evidence suggests reactive oxygen species play a vital role in programmed cell death, stress responses, plant defense against pathogens and systemic stress signaling in conjunction with antioxidant production. Here, we propose that reactive oxygen species and antioxidants, as both universal and evolutionarily conserved, are likely to play important role(s) in symbiotic interactions. To support this hypothesis we review the root and foliar fungal endophyte literature specific to fungal-plant symbiotum production of reactive oxygen species and antioxidants in response to stress. These asymptomatic fungi can produce antioxidants in response to both biotic and abiotic stress when grown in culture as well as in planta. In addition, there is a growing but nascent literature reporting a significant impact of endophyte colonization on the antioxidant activity of colonized (E+) hosts when compared to uncolonized (E-) hosts, especially when exposed to stress. Here we summarize general patterns emerging from the growing literature specific to antioxidant activity of endophytes in colonized hosts and bring up possible future research questions and approaches. The consequences of changes in reactive oxygen species production and increased antioxidant activity in the symbiotum appear to be beneficial in many instances; but costs are also indicated. Unexplored questions are: 1) to what extent do antioxidants originating from the fungal endophyte mediate host metabolism, and thereby control host responses to endophyte colonization; (2) what role do fungal, plant, or symbiotum produced reactive oxygen species and antioxidants have in determining symbiotic outcome between extremes of pathogenicity and mutualism; and (3) what role if any, do the production of reactive oxygen species and their antioxidant counterparts play in the symbiotum’s ability to respond to changing selection pressures? If as the literature suggests, such endophyte imposed mediation can be utilized to foster increases in plant production in resource limited habitats then the utilization of fungal endophytes may prove useful in agronomic and conservation settings.

Chemical Ecology Mediated by Fungal Endophytes in Grasses

Defensive mutualism is widely accepted as providing the best framework for understanding how seed-transmitted, alkaloid producing fungal endophytes of grasses are maintained in many host populations. Here, we first briefly review current knowledge of bioactive alkaloids produced by systemic grass-endophytes. New findings suggest that chemotypic diversity of the endophyte-grass symbiotum is far more complex, involving multifaceted signaling and chemical cross-talk between endophyte and host cells (e.g., reactive oxygen species and antioxidants) or between plants, herbivores, and their natural enemies (e.g., volatile organic compounds, and salicylic acid and jasmonic acid pathways). Accumulating evidence also suggests that the tight relationship between the systemic endophyte and the host grass can lead to the loss of grass traits when the lost functions, such as plant defense to herbivores, are compensated for by an interactive endophytic fungal partner. Furthermore, chemotypic diversity of a symbiotum appears to depend on the endophyte and the host plant life histories, as well as on fungal and plant genotypes, abiotic and biotic environmental conditions, and their interactions. Thus, joint approaches of (bio)chemists, molecular biologists, plant physiologists, evolutionary biologists, and ecologists are urgently needed to fully understand the endophyte-grass symbiosis, its coevolutionary history, and ecological importance. We propose that endophyte-grass symbiosis provides an excellent model to study microbially mediated multirophic interactions from molecular mechanisms to ecology.

Neotyphodium endophyte infection frequency in annual grass populations: relative importance of mutualism and transmission efficiency

Persistence and ubiquity of vertically transmitted Neotyphodium endophytes in grass populations is puzzling because infected plants do not consistently exhibit increased fitness. Using an annual grass population model, we show that the problems for matching endophyte infection and mutualism are likely to arise from difficulties in detecting small mutualistic effects, variability in endophyte transmission efficiency and an apparent prevalence of non-equilibrium in the dynamics of infection. Although endophytes would ultimately persist only if the infection confers some fitness increase to the host plants, such an increase can be very small, as long as the transmission efficiency is sufficiently high. In addition, imperfect transmission limits effectively the equilibrium infection level if the infected plants exhibit small or large reproductive advantage. Under frequent natural conditions, the equilibrium infection level is very sensitive to small changes in transmission efficiency and host reproductive advantage, while convergence to such an equilibrium is slow. As a consequence, seed immigration and environmental fluctuation are likely to keep local infection levels away from equilibrium. Transient dynamics analysis suggests that, when driven by environmental fluctuation, infection frequency increases would often be larger than decreases. By contrast, when due to immigration, overrepresentation of infected individuals tends to vanish faster than equivalent overrepresentation of non-infected individuals.

Evolution of Glyphosate-Resistant Johnsongrass (Sorghum halepense) in Glyphosate-Resistant Soybean

Abstract In Argentinean crop fields, weed control is mainly achieved by intense use of glyphosate as a nonselective and/or selective herbicide. Glyphosate use is very high as more than 95% of the 16 million ha soybean crop consists of glyphosate-resistant cultivars, always treated with this herbicide. From initial success, inconsistent glyphosate control of Johnsongrass, an invading C4 perennial grass of soybean crops, has become evident to producers from northern Argentina over the last 3 yr. Prior to this, glyphosate provided good control. This study evaluated the nature of these recurrent glyphosate failures in Johnsongrass. Experiments conducted with Johnsongrass plants obtained from seed and rhizome phytomers collected from fields with intense glyphosate use history showed that these populations showed differential survival and biomass productivity when glyphosate treated than Johnsongrass plants obtained from similar propagules collected from field sites with no history of glyphosate use. This empirical evidence establishes that the Johnsongrass survival in glyphosate-treated transgenic soybean fields from northern Argentina is due to evolved glyphosate resistance. Nomenclature: Glyphosate; Johnsongrass, Sorghum halepense L. Pers. SORHA; soybean, Glycine max (L.) Merr

Imperfect Vertical Transmission of the Endophyte Neotyphodium in Exotic Grasses in Grasslands of the Flooding Pampa

Cool-season grasses establish symbioses with vertically transmitted Neotyphodium endophytes widespread in nature. The frequency of endophyte-infected plants in closed populations (i.e., without migrations) depends on both the differential fitness between infected and non-infected plants, and the endophyte-transmission efficiency. Most studies have been focused on the first mechanism ignoring the second. Infection frequency and endophyte transmission from vegetative tissues to seeds were surveyed in two grasses growing in vegetation units that differ in flood and grazing regimes, and soil salinity. Transmission efficiency and infection frequency for tall fescue did not vary significantly and were 0.98 and 1.00, respectively. For Italian ryegrass, transmission efficiency and infection frequency were 0.88 and 0.57 in humid prairies, and 0.96 and 0.96 in the other vegetation units. Only in humid mesophytic meadows, the observed pattern was irrespective of the presence or absence of grazers. Our results suggest that selection forces for endophyte infection are different for both species. Imperfect transmission was only compensated in tall fescue through an increased fitness of infected plants. Interpreting variations of infection frequency only in terms of differential fitness can be misleading, considering that endophyte transmission can be imperfect and variable in nature. Therefore, this study highlights the importance of measuring transmission efficiency.

Glyphosate resistance in Sorghum halepense and Lolium rigidum is reduced at suboptimal growing temperatures

BACKGROUND Glyphosate resistance in populations of the C(4) perennial Sorghum halepense (Johnsongrass) and C(3) annual Lolium rigidum (rigid ryegrass) has evolved and been documented in many cropping areas around the globe. In S. halepense and in the majority of reported cases in L. rigidum the glyphosate resistance trait has been associated with a mechanism that reduces glyphosate translocation within plants. Here, the significant decrease in the glyphosate resistance level when resistant plants of S. halepense and L. rigidum are grown at suboptimal cool temperature conditions is reported. RESULTS Lowering temperature from 30 to 19 °C in S. halepense and from 19 to 8 °C in L. rigidum significantly reduced both plant survival and above-ground biomass produced by glyphosate-resistant plants. Thus, glyphosate resistance parameters significantly decreased when glyphosate-treated resistant plants of both species were grown under non-optimal temperature conditions. The results suggest that the resistance mechanism against glyphosate damage is less efficient at sub-optimal [corrected] growing temperatures. CONCLUSION It is possible to increase the control of glyphosate-resistant S. halepense and L. rigidum populations by treatment with glyphosate during growing conditions at suboptimal low temperatures. Conversely, glyphosate failure will continue to occur on glyphosate-resistant populations treated during periods of higher temperatures

Experimental Methods for Estimation of Plant Fitness Costs Associated with Herbicide-Resistance Genes

Herbicide Resistance Genes andPlant FitnessSince the beginning of agriculture, crops havebeen exposed to recurrent invasion by weeds thatcan impose severe reductions in crop quality andyield. There have been continuing efforts to reducethe impacts of weeds on production. More than40 yr ago, overreliance on herbicide technology toreduce weed infestations resulted in the selection ofadaptive traits that enabled weed survival andreproduction under herbicide treatments (De´lye etal. 2007; Powles and Yu 2010; Vila-Aiub et al.2008). As a result, herbicide resistance in . 200weed species has evolved worldwide (Heap 2013;Powles 2008).Resistant weeds are able to withstand the toxicityof herbicides because of the presence of resistancealleles originating from random DNA mutations(Powles and Yu 2010). These resistance allelesregulate a number of highly efficient, constitutivedefense mechanisms that prevent herbicides frominhibiting key metabolic pathways. A set of defensemechanisms are involved in a reduction in herbicidethat reaches the herbicide target protein (nontarget-site resistance) (Powles and Yu 2010). Mechanismsthat (1) impair herbicide leaf uptake or transloca-tion within plants via vacuolar sequestration orreduced cellular uptake, or (2) change the chemicalproperties of herbicides via herbicide-enhancedmetabolism (detoxification) are included in thisgroup (Ge et al. 2012; Nandula et al. 2008; Prestonand Wakelin 2008; Preston et al. 1996; Sammonset al. 2010; Vila-Aiub et al. 2012; Wakelin et al.2004).Another type of defense mechanism againstherbicides involves a structural modification, viachanges in the amino acid sequence, of the herbicidetarget protein, which minimizes herbicide binding(target-site resistance) (Powles and Yu 2010). Geneoverexpression, resulting in the synthesis of excessiveherbicide sensitive target protein because of pro-moter changes or gene amplification, is alsoregarded a target-site resistance defense mechanism(Dinelli et al. 2006; Gaines et al. 2010).Target and nontarget-site herbicide resistancealleles protect plants from fatal damage caused byherbicides. In other words, the presence of resistancealleles minimizes the plant fitness reduction expect-ed from herbicide activity. Fitness can be defined asthe average success in producing offspring contrib-uting to the next generation by a particularphenotype relative to another phenotype (Crawley1997; Primack and Hyesoon 1989; Scott et al.2006). A simple way to represent plant fitness (W)is given in Futuyma (2013):W~P

Mutualism effectiveness and vertical transmission of symbiotic fungal endophytes in response to host genetic background.

Certain species of the Pooideae subfamily develop stress tolerance and herbivory resistance through symbiosis with vertically transmitted, asexual fungi. This symbiosis is specific, and genetic factors modulate the compatibility between partners. Although gene flow is clearly a fitness trait in allogamous grasses, because it injects hybrid vigor and raw material for evolution, it could reduce compatibility and thus mutualism effectiveness. To explore the importance of host genetic background in modulating the performance of symbiosis, Lolium multiflorum plants, infected and noninfected with Neotyphodium occultans, were crossed with genetically distant plants of isolines (susceptible and resistant to diclofop‐methyl herbicide) bred from two cultivars and exposed to stress. The endophyte improved seedling survival in genotypes susceptible to herbicide, while it had a negative effect on one of the genetically resistant crosses. Mutualism provided resistance to herbivory independently of the host genotype, but this effect vanished under stress. While no endophyte effect was observed on host reproductive success, it was increased by interpopulation plant crosses. Neither gene flow nor herbicide had an important impact on endophyte transmission. Host fitness improvements attributable to gene flow do not appear to result in direct conflict with mutualism while this seems to be an important mechanism for the ecological and contemporary evolution of the symbiotum.

The interplay between the effectiveness of the grass-endophyte mutualism and the genetic variability of the host plant

Neotyphodium endophytic fungi, the asexual state of Epichloë species, protect cool‐season grasses against stresses. The outcomes of Neotyphodium‐grass symbioses are agronomically relevant as they may affect the productivity of pastures. It has been suggested that the mutualism is characteristic of agronomic grasses and that differential rates of gene flow between both partners’ populations are expected to disrupt the specificity of the association and, thus, the mutualism in wild grasses. We propose that compatibility is necessary but not sufficient to explain the outcomes of Neotyphodium‐grass symbiosis, and advance a model that links genetic compatibility, mutualism effectiveness, and endophyte transmission efficiency. For endophytes that reproduce clonally and depend on allogamous hosts for reproduction and dissemination, we propose that this symbiosis works as an integrated entity where gene flow promotes its fitness and evolution. Compatibility between the host plant and the fungal endophyte would be high in genetically close parents; however, mutualism effectiveness and transmission efficiency would be low in fitness depressed host plants. Increasing the genetic distance of mating parents would increase mutualism effectiveness and transmission efficiency. This tendency would be broken when the genetic distance between parents is high (out‐breeding depression). Our model allows for testable hypotheses that would contribute to understand the coevolutionary origin and future of the endophyte‐grass mutualism.

Competing neighbors: light perception and root function

Plant responses to competition have often been described as passive consequences of reduced resource availability. However, plants have mechanisms to forage for favorable conditions and anticipate competition scenarios. Despite the progresses made in understanding the role of light signaling in modulating plant–plant interactions, little is known about how plants use and integrate information gathered by their photoreceptors aboveground to regulate performance belowground. Given that the phytochrome family of photoreceptors plays a key role in the acquisition of information about the proximity of neighbors and canopy cover, it is tempting to speculate that changes in the red:far-red (R:FR) ratio perceived by aboveground plant parts have important implications shaping plant behavior belowground. Exploring data from published experiments, we assess the neglected role of light signaling in the control of root function. The available evidence indicates that plant exposure to low R:FR ratios affects root growth and morphology, root exudate profiles, and interactions with beneficial soil microorganisms. Although dependent on species identity, signals perceived aboveground are likely to affect root-to-root interactions. Root systems could also be guided to deploy new growth predominantly in open areas by light signals perceived by the shoots. Studying interactions between above- and belowground plant–plant signaling is expected to improve our understanding of the mechanisms of plant competition.