Unraveling root and rhizosphere traits in temperate maize landraces and modern cultivars: Implications for soil resource acquisition and drought adaptation.

A holistic understanding of plant strategies to acquire soil resources is pivotal in achieving sustainable food security. However, we lack knowledge about variety-specific root and rhizosphere traits for resource acquisition, their plasticity and adaptation to drought. We conducted a greenhouse experiment to phenotype root and rhizosphere traits (mean root diameter [Root D], specific root length [SRL], root tissue density, root nitrogen content, specific rhizosheath mass [SRM], arbuscular mycorrhizal fungi [AMF] colonization) of 16 landraces and 22 modern cultivars of temperate maize (Zea mays L.). Our results demonstrate that landraces and modern cultivars diverge in their root and rhizosphere traits. Although landraces follow a 'do-it-yourself' strategy with high SRLs, modern cultivars exhibit an 'outsourcing' strategy with increased mean Root Ds and a tendency towards increased root colonization by AMF. We further identified that SRM indicates an 'outsourcing' strategy. Additionally, landraces were more drought-responsive compared to modern cultivars based on multitrait response indices. We suggest that breeding leads to distinct resource acquisition strategies between temperate maize varieties. Future breeding efforts should increasingly target root and rhizosphere economics, with SRM serving as a valuable proxy for identifying varieties employing an outsourcing resource acquisition strategy.

Keep in touch: the soil-root hydraulic continuum and its role in drought resistance in crops.

Drought is a major threat to food security worldwide. Recently, the root-soil interface has emerged as a major site of hydraulic resistance during water stress. Here, we review the impact of soil drying on whole-plant hydraulics and discuss mechanisms by which plants can adapt by modifying the properties of the rhizosphere either directly or through interactions with the soil microbiome.

Soil phosphorus availability alters the correlations between root phosphorus-uptake rates and net photosynthesis of dominant C3 and C4 species in a typical temperate grassland of Northern China.

Phosphorus (P) fertilization can alleviate a soil P deficiency in grassland ecosystems. Understanding plant functional traits that enhance P uptake can improve grassland management. We measured impacts of P addition on soil chemical and microbial properties, net photosynthetic rate (Pn ) and nonstructural carbohydrate concentrations ([NSC]), and root P-uptake rate (PUR), morphology, anatomy, and exudation of two dominant grass species: Leymus chinensis (C3 ) and Cleistogenes squarrosa (C4 ). For L. chinensis, PUR and Pn showed a nonlinear correlation. Growing more adventitious roots compensated for the decrease in P transport per unit root length, so that it maintained a high PUR. For C. squarrosa, PUR and Pn presented a linear correlation. Increased Pn was associated with modifications in root morphology, which further enhanced its PUR and a greater surplus of photosynthate and significantly stimulated root exudation (proxied by leaf [Mn]), which had a greater impact on rhizosheath micro-environment and microbial PLFAs. Our results present correlations between the PUR and the Pn of L. chinensis and C. squarrosa and reveal that NSC appeared to drive the modifications of root morphology and exudation; they provide more objective basis for more efficient P-input in grasslands to address the urgent problem of P deficiency.

Above and belowground traits impacting transpiration decline during soil drying in 48 maize (Zea mays) genotypes.

BACKGROUND AND AIMS: Stomatal regulation allows plants to promptly respond to water stress. However, our understanding of the impact of above and belowground hydraulic traits on stomatal regulation remains incomplete. The objective of this study was to investigate how key plant hydraulic traits impact transpiration of maize during soil drying. We hypothesize that the stomatal response to soil drying is related to a loss in soil hydraulic conductivity at the root-soil interface, which in turn depends on plant hydraulic traits. METHODS: We investigate the response of 48 contrasting maize (Zea mays) genotypes to soil drying, utilizing a novel phenotyping facility. In this context, we measure the relationship between leaf water potential, soil water potential, soil water content and transpiration, as well as root, rhizosphere and aboveground plant traits. KEY RESULTS: Genotypes differed in their responsiveness to soil drying. The critical soil water potential at which plants started decreasing transpiration was related to a combination of above and belowground traits: genotypes with a higher maximum transpiration and plant hydraulic conductance as well as a smaller root and rhizosphere system closed stomata at less negative soil water potentials. CONCLUSIONS: Our results demonstrate the importance of belowground hydraulics for stomatal regulation and hence drought responsiveness during soil drying. Furthermore, this finding supports the hypothesis that stomata start to close when soil hydraulic conductivity drops at the root-soil interface.

Effects of Soil Water Shortage on Seedling Shoot and Root Growth of Saragolle Lucana Tetraploid Wheat (Triticum durum Desf.) Landrace.

Ancient wheats may be a source of traits that are useful for the tolerance of climate change foreseen conditions of raising temperatures and low water availability. Previous research has shown a fine root system and a high mass of rhizosheath per unit root mass in the italian durum wheat (Triticum durum Desf) landrace Saragolle Lucana, and this may be relevant for successfully facing adverse conditions during seedling establishment. We investigated the effect of soil water shortage in Saragolle seedlings on root architecture, rhizosheath formation and biomass allocation. Pot experiments were conducted by comparing two levels of soil available water content (AWC): WW (100% of AWC) and DS (50% of AWC). Phenology was delayed by eight days in DS and above and belowground traits were measured at Zadoks 1.3 for each treatment. Biometric data collected at the same phenological stage show that DS plants did not reach the levels of biomass, surface area and space occupation of WW even after attaining the same developmental stage. Namely, plant dimensions were lower at low soil water availability, with the exception of rhizosheath production: DS yielded a 50% increase in rhizosheath mass and 32% increase in rhizosheath mass per unit root mass. The proportion of plant mass reduction in DS was 29.7% for aboveground parts and 34.7% for roots, while reductions in leaf and root surface areas exceeded 43%. The root/shoot mass and area ratios were not significantly different between treatments, and a higher impact on aboveground than on belowground traits at reduced available water was shown only by a lower ratio of shoot height to root depth in DS than in WW. Increases in rhizosheath in absolute and relative terms, which were observed in our experiment in spite of smaller root systems in the ancient durum wheat variety Saragolle lucana at DS, may provide an interesting trait for plant performance in conditions of low soil water availability both for water-related issue and for other effects on plant nutrition and relations with the rhizosphere.

Rhizosheath-root system changes exopolysaccharide content but stabilizes bacterial community across contrasting seasons in a desert environment.

BACKGROUND: In hot deserts daily/seasonal fluctuations pose great challenges to the resident organisms. However, these extreme ecosystems host unique microenvironments, such as the rhizosheath-root system of desert speargrasses in which biological activities and interactions are facilitated by milder conditions and reduced fluctuations. Here, we examined the bacterial microbiota associated with this structure and its surrounding sand in the desert speargrass Stipagrostis pungens under the contrasting environmental conditions of summer and winter in the Sahara Desert. RESULTS: The belowground rhizosheath-root system has higher nutrient and humidity contents, and cooler temperatures than the surrounding sand. The plant responds to the harsh environmental conditions of the summer by increasing the abundance and diversity of extracellular polymeric substances (EPS) compared to the winter. On the contrary, the bacterial community associated with the rhizosheath-root system and its interactome remain stable and, unlike the bulk sand, are unaffected by the seasonal environmental variations. The rhizosheath-root system bacterial communities are consistently dominated by Actinobacteria and Alphaproteobacteria and form distinct bacteria communities from those of bulk sand in the two seasons. The microbiome-stabilization mediated by the plant host acts to consistently retain beneficial bacteria with multiple plant growth promoting functions, including those capable to produce EPS, which increase the sand water holding capacity ameliorating the rhizosheath micro-environment. CONCLUSIONS: Our results reveal the capability of plants in desert ecosystems to stabilize their below ground microbial community under seasonal contrasting environmental conditions, minimizing the heterogeneity of the surrounding bulk sand and contributing to the overall holobiont resilience under poly-extreme conditions.

Interactive effects of phosphorus fertilization and salinity on plant growth, phosphorus and sodium status, and tartrate exudation by roots of two alfalfa cultivars.

BACKGROUND AND AIMS: Soil phosphorus (P) deficiency and salinity are constraints to crop productivity in arid and semiarid regions. Salinity may weaken the effect of P fertilization on plant growth. We investigated the interactive effects of soil P availability and salinity on plant growth, P nutrition and salt tolerance of two alfalfa (Medicago sativa) cultivars. METHODS: A pot experiment was carried out to grow two cultivars of alfalfa in a loess soil under a combination of different rates of added P (0, 40, 80 and 160 mg P kg-1 soil as monopotassium phosphate) and sodium chloride (0, 0.4, 0.8 and 1.6 g NaCl kg-1 soil). Plant biomass, concentrations of P ([P]), sodium ([Na]) and potassium ([K]) were determined, and rhizosheath carboxylates were analysed. KEY RESULTS: There were significant interactions between soil P availability and salinity on some, but not all, of the parameters investigated, and interactions depended on cultivar. Plant growth and P uptake were enhanced by P fertilization, but inhibited by increased levels of salinity. Increasing the salinity resulted in decreased plant P-uptake efficiency and [K]/[Na]. Only soil P availability had a significant effect on the amount of tartrate in the rhizosheath of both cultivars. CONCLUSIONS: Increased salinity aggravated P deficiency. Appropriate application of P fertilizers improved the salt tolerance of alfalfa and increased its productivity in saline soils.

Diversity and function of microbial communities in the sand sheath of Agropyron cristatum by metagenomic analysis.

The roots of most gramineous plants are surrounded by a variety of microorganisms; however, few studies have focused on the rhizosheath of psammophytes. Therefore, in this study, we used Illumina HiSeq high-throughput sequencing technology to analyse the composition and functional diversity of microbial communities in the rhizosheath of sand-grown Agropyron cristatum (L.) Gaertn. We found that the number of species and functions of microbial communities gradually decreased from the rhizosheath to the bulk soil. Thus, the microbial composition of the rhizosheath was richer and more diverse, and the abundance of bacteria, including Sphingosinicella, Rhizorhabdus, Friedmanniella, Geodermatophilus, Blastococcus, and Oscillatoria, was higher, and the abundance of fungi, such as Mycothermus, was higher. The abundance of CO2 fixation-related genes (acsA, Pcc, and cbbL) in the carbon cycle; NO3-, NO2-, NH2OH, and N2 transformation genes (nrtP, nirS, hao, and nifK) in the nitrogen cycle; soxB/A/C, Sat, and dsrB genes in the sulphur cycle; and 1-phosphate mannitol dehydrogenase (MtlD) gene and polyketide synthase gene (pks) were higher in the rhizosheath than in the bulk soil, as well as genes related to phosphorus uptake in the phosphorus cycle. Our findings showed that the rhizosheath may host the predominant microbial species related to the formation of a rhizosheath.

Abscisic Acid Mediates Drought-Enhanced Rhizosheath Formation in Tomato.

The rhizosheath, commonly defined as soil adhering to the root surface, may confer drought tolerance in various crop species by enhancing access to water and nutrients under drying stress conditions. Since the role of phytohormones in establishing this trait remains largely unexplored, we investigated the role of ABA in rhizosheath formation of wild-type (WT) and ABA-deficient (notabilis, not) tomatoes. Both genotypes had similar rhizosheath weight, root length, and root ABA concentration in well-watered soil. Drying stress treatment decreased root length similarly in both genotypes, but substantially increased root ABA concentration and rhizosheath weight of WT plants, indicating an important role for ABA in rhizosheath formation. Neither genotype nor drying stress treatment affected root hair length, but drying stress treatment decreased root hair density of not. Under drying stress conditions, root hair length was positively correlated with rhizosheath weight in both genotypes, while root hair density was positively correlated with rhizosheath weight in well-watered not plants. Root transcriptome analysis revealed that drought stress increased the expression of ABA-responsive transcription factors, such as AP2-like ER TF, alongside other drought-regulatory genes associated with ABA (ABA 8'-hydroxylase and protein phosphatase 2C). Thus, root ABA status modulated the expression of specific gene expression pathways. Taken together, drought-induced rhizosheath enhancement was ABA-dependent, but independent of root hair length.

Drought stress and plant ecotype drive microbiome recruitment in switchgrass rhizosheath.

The rhizosheath, a layer of soil grains that adheres firmly to roots, is beneficial for plant growth and adaptation to drought environments. Switchgrass is a perennial C4 grass which can form contact rhizosheath under drought conditions. In this study, we characterized the microbiomes of four different rhizocompartments of two switchgrass ecotypes (Alamo and Kanlow) grown under drought or well-watered conditions via 16S ribosomal RNA amplicon sequencing. These four rhizocompartments, the bulk soil, rhizosheath soil, rhizoplane, and root endosphere, harbored both distinct and overlapping microbial communities. The root compartments (rhizoplane and root endosphere) displayed low-complexity communities dominated by Proteobacteria and Firmicutes. Compared to bulk soil, Cyanobacteria and Bacteroidetes were selectively enriched, while Proteobacteria and Firmicutes were selectively depleted, in rhizosheath soil. Taxa from Proteobacteria or Firmicutes were specifically selected in Alamo or Kanlow rhizosheath soil. Following drought stress, Citrobacter and Acinetobacter were further enriched in rhizosheath soil, suggesting that rhizosheath microbiome assembly is driven by drought stress. Additionally, the ecotype-specific recruitment of rhizosheath microbiome reveals their differences in drought stress responses. Collectively, these results shed light on rhizosheath microbiome recruitment in switchgrass and lay the foundation for the improvement of drought tolerance in switchgrass by regulating the rhizosheath microbiome.

Phosphorus uptake is associated with the rhizosheath formation of mature cluster roots in white lupin under soil drying and phosphorus deficiency.

Phosphorus (P) deficiency largely restricts plant growth and lead to severe yield losses. Therefore, identification of novel root traits to improve P uptake is needed to circumvent yield losses. White lupin (Lupinus albus) is a legume crop that develops cluster roots and has the high phosphorus use efficiency in low P soils. We aimed to investigate the association between cluster roots (CR) rhizosheath formation and P uptake in white lupin. Rhizosheath formation and P concentration were evaluated under four soil treatments. CR increased up to 2.5-fold of overall plant dry weight under SD-P compared to WW + P (control), partly attributable to variations in CR development. Our data showed that SD-P significantly increase rhizosheath weight in white lupin. Among the root segments, MCR showed improved P accumulation in the root which is associated with increased MCR rhizosheath weight. Additionally, a positive correlation was observed between MCR rhizosheath weight and P uptake. Moreover, high sucrose content was recorded in MCR, which may contribute in CR growth under SD-P. Expression analysis of genes related to sucrose accumulation (LaSUC1, LaSUC5, and LaSUC9) and phosphorus uptake (LaSPX3, LaPHO1, and LaPHT1) exhibited peaked expression in MCR under SD-P. This indicate that root sucrose status may facilitate P uptake under P starvation. Together, the ability to enhance P uptake of white lupin is largely associated with MCR rhizosheath under SD-P. Our results showed that gene expression modulation of CR forming plant species, demonstrating that these novel root structures may play crucial role in P acquisition from the soil. Our findings could be implicated for developing P and water efficient crop via CR development in sustainable agriculture.

Rhizosheath microbes induce root immune response under soil drying.

The rhizosheath is an important drought-adaptive trait in roots of many angiosperms and has been regarded as a potential trait for future agricultural sustainability. In recent studies, we found that rice roots could form a pronounced rhizosheath under moderate soil drying (MSD) but not under continuous flooding irrigation (CF). The formation of rhizosheaths substantially changes the microbial community structure in endosphere root tissues and the rhizosphere in rice, which may induce a plant immune response. However, the manner by which the formation of rhizosheaths regulates the immune system of roots remains largely unknown. Here, we have analyzed the root transcriptomes of drought-tolerant rice and drought-sensitive rice under both MSD (rhizosheath-root) and CF (root without rhizosheath) conditions. Our results suggest that rhizosheath-associated microbes may trigger plant immune pathways in root under MSD, including the first line of defense component pattern-triggered immunity and the second line of defense component effector-triggered immunity. These data expand our understanding of rhizosheath-associated microbes and plant interactions.

Exploring the structural changes in nitrogen-fixing microorganisms of rhizosheath during the growth of Stipagrostis pennata in the desert.

PURPOSE: Rhizosheath is an adaptive feature for the survival of Stipagrostis pennata in desert systems. Although microorganisms play important ecological roles in promoting the nitrogen cycle of rhizosheath, the diversity and function of nitrogen-fixing microorganism communities have not been fully understood. MATERIALS AND METHODS: Therefore, the aim of the present study is to explore the nitrogen fixation ability of rhizosheaths and the changes in abundance of nitrogen-fixing microorganisms at different growth periods of S. pennata. We sequenced the nifH gene through sequencing to identify the structure and diversity of nitrogen-fixing microorganisms of S. pennata at different growth periods of rhizosheaths. RESULTS: A total of 1256 operational taxonomic units (OTUs) were identified by nifH sequence and distributed in different growth periods. There were five OTUs distributed in each sample at the same time, and the abundance and diversity of microorganisms in fruit period were much higher than those in other periods. Mainly four phyla were involved, among which Proteobacteria was the most abundant in all groups. CONCLUSIONS: In general, the present study investigated the abundance and characteristics of nitrogen-fixing microorganisms of rhizosheaths in different growth periods of S. pennata. It also may elucidate and indicate that the structure of nitrogen-fixing microorganisms of rhizosheaths in different growth periods of S. pennata had changed.

Abscisic acid mediates barley rhizosheath formation under mild soil drying by promoting root hair growth and auxin response.

Soil drying enhances root ABA accumulation and rhizosheath formation, but whether ABA mediates rhizosheath formation is unclear. Here, we used the ABA-deficient mutant Az34 to investigate molecular and morphological changes by which ABA could affect rhizosheath formation. Mild soil drying with intermittent watering increased rhizosheath formation by promoting root and root hair elongation. Attenuated root ABA accumulation in Az34 barley constrained the promotion of root length and root hair length by drying soil, such that Az34 had a smaller rhizosheath. Pharmacological experiments of adding fluridone (an ABA biosynthesis inhibitor) and ABA to drying soil restricted and enhanced rhizosheath formation respectively in Az34 and wild-type Steptoe barley. RNA sequencing suggested that ABA accumulation mediates auxin synthesis and responses and root and root hair elongation in drying soil. In addition, adding indole-3-acetic acid (IAA) to drying soil increased rhizosheath formation by promoting root and root hair elongation in Steptoe and Az34 barley. Together, these results show that ABA accumulation induced by mild soil drying enhance barley rhizosheath formation, which may be achieved through promoting auxin response.

Root hairs are the most important root trait for rhizosheath formation of barley (Hordeum vulgare), maize (Zea mays) and Lotus japonicus (Gifu).

BACKGROUND AND AIMS: Rhizosheaths are defined as the soil adhering to the root system after it is extracted from the ground. Root hairs and mucilage (root exudates) are key root traits involved in rhizosheath formation, but to better understand the mechanisms involved their relative contributions should be distinguished. METHODS: The ability of three species [barley (Hordeum vulgare), maize (Zea mays) and Lotus japonicus (Gifu)] to form a rhizosheath in a sandy loam soil was compared with that of their root-hairless mutants [bald root barley (brb), maize root hairless 3 (rth3) and root hairless 1 (Ljrhl1)]. Root hair traits (length and density) of wild-type (WT) barley and maize were compared along with exudate adhesiveness of both barley and maize genotypes. Furthermore, root hair traits and exudate adhesiveness from different root types (axile versus lateral) were compared within the cereal species. KEY RESULTS: Per unit root length, rhizosheath size diminished in the order of barley > L. japonicus > maize in WT plants. Root hairs significantly increased rhizosheath formation of all species (3.9-, 3.2- and 1.8-fold for barley, L. japonicus and maize, respectively) but there was no consistent genotypic effect on exudate adhesiveness in the cereals. While brb exudates were more and rth3 exudates were less adhesive than their respective WTs, maize rth3 bound more soil than barley brb. Although both maize genotypes produced significantly more adhesive exudate than the barley genotypes, root hair development of WT barley was more extensive than that of WT maize. Thus, the greater density of longer root hairs in WT barley bound more soil than WT maize. Root type did not seem to affect rhizosheath formation, unless these types differed in root length. CONCLUSIONS: When root hairs were present, greater root hair development better facilitated rhizosheath formation than root exudate adhesiveness. However, when root hairs were absent root exudate adhesiveness was a more dominant trait.

Specific Rhizobacteria Responsible in the Rhizosheath System of Kengyilia hirsuta.

The rhizosheath is a critical interface supporting the exchange of resources between plants and their associated environment of soil. Favorable microenvironment of rhizosphere soil provides the rhizosheath formed and then promotes desert plant survival. However, it remains unclear how rhizosheath benefits the colonization of pioneer plants in alpine desert under changing environment. In this study, we investigated the effect of different soil moisture and sterilization treatments (three moisture levels and unsterilized or sterilized soil) on rhizosheath forming process of Kengyilia hirsuta (K. hirsuta), a sand-inhabiting and drought-resistant pioneer plant of the Tibetan Plateau desert. The results showed that in both unsterilized and sterilized soil, increasing soil moisture first increased and then decreased rhizosheath weight, with the highest value is 25%. During rhizosheath formation, developing rhizosheaths were selectively enriched in the bacterial genera Massilia and Arthrobacter. These suggest the existence of a highly specialized signal recognition system during rhizosheath formation that involves the accumulation of bacteria. These bacterial species exhibited different roles in the process of rhizosheath formation and is an advantageous strategy for K. hirsuta.

Significance of root hairs for plant performance under contrasting field conditions and water deficit.

BACKGROUND AND AIMS: Previous laboratory studies have suggested selection for root hair traits in future crop breeding to improve resource use efficiency and stress tolerance. However, data on the interplay between root hairs and open-field systems, under contrasting soils and climate conditions, are limited. As such, this study aims to experimentally elucidate some of the impacts that root hairs have on plant performance on a field scale. METHODS: A field experiment was set up in Scotland for two consecutive years, under contrasting climate conditions and different soil textures (i.e. clay loam vs. sandy loam). Five barley (Hordeum vulgare) genotypes exhibiting variation in root hair length and density were used in the study. Root hair length, density and rhizosheath weight were measured at several growth stages, as well as shoot biomass, plant water status, shoot phosphorus (P) accumulation and grain yield. KEY RESULTS: Measurements of root hair density, length and its correlation with rhizosheath weight highlighted trait robustness in the field under variable environmental conditions, although significant variations were found between soil textures as the growing season progressed. Root hairs did not confer a notable advantage to barley under optimal conditions, but under soil water deficit root hairs enhanced plant water status and stress tolerance resulting in a less negative leaf water potential and lower leaf abscisic acid concentration, while promoting shoot P accumulation. Furthermore, the presence of root hairs did not decrease yield under optimal conditions, while root hairs enhanced yield stability under drought. CONCLUSIONS: Selecting for beneficial root hair traits can enhance yield stability without diminishing yield potential, overcoming the breeder's dilemma of trying to simultaneously enhance both productivity and resilience. Therefore, the maintenance or enhancement of root hairs can represent a key trait for breeding the next generation of crops for improved drought tolerance in relation to climate change.

Moderate water stress in rice induces rhizosheath formation associated with abscisic acid and auxin responses.

The rhizosheath is known to be beneficial for drought resistance in many plants, but the regulation of rhizosheath formation in rice plants is unclear. Here, we investigate rhizosheath formation in different rice varieties and root hair mutants. Our results showed that moderate water stress in rice induced rhizosheath formation. The soil porosity and water content were higher in the rice rhizosheath than in the rice bulk soil under moderate water stress. Additionally, rhizosheath formation in short root hair mutants was lower than in wild-type rice under moderate water stress. Moreover, transcriptomic results indicated that abscisic acid (ABA) and auxin were involved in root and root hair responses in rhizosheath formation. Further, blocking ABA and auxin pathways in wild type and in rhl1-1, the shortest root hair mutant, rhizosheath formation and root hair length were significantly decreased under moderate water stress. However, wild type plants maintained a higher root ABA content, root basipetal auxin transport, root hair length, and amount of rhizosheath than did rhl1-1. Our results suggest that moderate water stress in rice induces rhizosheath formation by modulating the ABA and auxin responses to regulate root and root hair growth, which may be used to breed rice varieties resistant to drought.

Comparative metabolite profiling of two switchgrass ecotypes reveals differences in drought stress responses and rhizosheath weight.

MAIN CONCLUSION: Rhizosheath comprises soil that adheres firmly to roots. In this study, two ecotypes of switchgrass with different rhizosheath sizes after drought stress were analyzed which showed metabolic differences under drought conditions. The rhizosheath comprises soil that adheres firmly to roots by a combination of root hairs and mucilage and may aid in root growth under soil drying. The aim of this work is to reveal the potential metabolites involved in rhizosheath formation under drought stress conditions. Panicum virgatum L. (switchgrass), which belongs to the Poaceae family, is an important biofuel and fodder crop in drought areas. Five switchgrass ecotypes (cv. Alamo, cv. Blackwake, cv. Summer, cv. Cave-in-Rock and cv. Kanlow) have a broad range of rhizosheath weight under drought conditions. For two selected ecotypes with contrast rhizosheath weight (cv. Alamo and cv. Kanlow), root hair length and density, lateral root number, root morphological parameters were measured, and real-time qRT-PCR was performed. Gas chromatography mass spectrophotometry (GC-MS) was used to determine the primary metabolites in the shoots and roots of selected ecotypes under drought stress conditions. The change trends of root hair length and density, lateral root number and related gene expression were consistent with rhizosheath weight in Alamo and Kanlow under drought and watered conditions. For root morphological parameters, Alamo grew deeper than Kanlow, while Kanlow exhibited higher values for other parameters. In this study, the levels of amino acids, sugars and organic acids were significantly changed in response to drought stress in two switchgrass ecotypes. Several metabolites including amino acids (arginine, isoleucine, methionine and cysteine) and sugars (kestose, raffinose, fructose, fucose, sorbose and xylose) in the large soil-sheathed roots of Alamo and Kanlow were significantly increased compared to small or no soil-sheathed roots of Alamo and Kanlow. Difference in rhizosheath size is reflected in the plant internal metabolites under drought stress conditions. Additionally, our results highlight the importance of using metabolite profiling and provide a better understanding of rhizosheath formation at the cellular level.

Rhizosheath microbial community assembly of sympatric desert speargrasses is independent of the plant host.

BACKGROUND: The rhizosheath-root system is an adaptive trait of sandy-desert speargrasses in response to unfavourable moisture and nutritional conditions. Under the deserts' polyextreme conditions, plants interact with edaphic microorganisms that positively affect their fitness and resistance. However, the trophic simplicity and environmental harshness of desert ecosystems have previously been shown to strongly influence soil microbial community assembly. We hypothesize that sand-driven ecological filtering constrains the microbial recruitment processes in the speargrass rhizosheath-root niche, prevailing over the plant-induced selection. METHODS: Bacterial and fungal communities from the rhizosheath-root compartments (endosphere root tissues, rhizosheath and rhizosphere) of three Namib Desert speargrass species (Stipagrostis sabulicola, S. seelyae and Cladoraphis spinosa) along with bulk sand have been studied to test our hypothesis. To minimize the variability determined by edaphic and climatic factors, plants living in a single dune were studied. We assessed the role of plant species vs the sandy substrate on the recruitment and selection, phylogenetic diversity and co-occurrence microbial networks of the rhizosheath-root system microbial communities. RESULTS: Microorganisms associated with the speargrass rhizosheath-root system were recruited from the surrounding bulk sand population and were significantly enriched in the rhizosheath compartments (105 and 104 of bacterial 16S rRNA and fungal ITS copies per gram of sand to up to 108 and 107 copies per gram, respectively). Furthermore, each rhizosheath-root system compartment hosted a specific microbial community demonstrating strong niche-partitioning. The rhizosheath-root systems of the three speargrass species studied were dominated by desert-adapted Actinobacteria and Alphaproteobacteria (e.g. Lechevalieria, Streptomyces and Microvirga) as well as saprophytic Ascomycota fungi (e.g. Curvularia, Aspergillus and Thielavia). Our results clearly showed a random phylogenetic turnover of rhizosheath-root system associated microbial communities, independent of the plant species, where stochastic factors drive neutral assembly. Co-occurrence network analyses also indicated that the bacterial and fungal community members of the rhizosheath-root systems established a higher number of interactions than those in the barren bulk sand, suggesting that the former are more stable and functional than the latter. CONCLUSION: Our study demonstrates that the rhizosheath-root system microbial communities of desert dune speargrasses are stochastically assembled and host-independent. This finding supports the concept that the selection determined by the desert sand prevails over that imposed by the genotype of the different plant species.