Different Responses of Soil Bacterial and Fungal Communities in Three Typical Vegetations following Nitrogen Deposition in an Arid Desert.

The effects of increased nitrogen (N) deposition on desert ecosystems have been extensively studied from a plant community perspective. However, the response of soil microbial communities, which play a crucial role in nutrient cycling, to N inputs and plant community types remains poorly understood. In this study, we conducted a two-year N-addition experiment with five gradients (0, 10, 30, 60, and 120 kg N ha-1 year-1) to evaluate the effect of increased N deposition on soil bacterial and fungal communities in three plant community types, namely, Alhagi sparsifolia Shap., Karelinia caspia (Pall.) Less. monocultures and their mixed community in a desert steppe located on the southern edge of the Taklimakan Desert, Northwest China. Our results indicate that N deposition and plant community types exerted an independent and significant influence on the soil microbial community. Bacterial α-diversity and community dissimilarity showed a unimodal pattern with peaks at 30 and 60 kg N ha-1 year-1, respectively. By contrast, fungal α-diversity and community dissimilarity did not vary significantly with increased N inputs. Furthermore, plant community type significantly altered microbial community dissimilarity. The Mantel test and redundancy analysis indicated that soil pH and total and inorganic N (NH4+ and NO3-) levels were the most critical factors regulating soil microbial communities. Similar to the patterns observed in taxonomic composition, fungi exhibit stronger resistance to N addition compared to bacteria in terms of their functionality. Overall, our findings suggest that the response of soil microbial communities to N deposition is domain-specific and independent of desert plant community diversity, and the bacterial community has a critical threshold under N enrichment in arid deserts.

Potential role of root-associated bacterial communities in adjustments of desert plant physiology to osmotic stress.

Plants possess the ability to adapt to osmotic stress by adjusting their physiology and morphology and by cooperating with their root-associated (rhizosphere and endosphere) microbial communities. However, the coordination of host self-regulation with root-associated microorganisms at the community level, especially for desert plants, remains unclear. This study investigated the morphophysiological responses of seedlings from the desert plant Alhagi sparsifolia Shap to osmotic stress, as well as the relationships between these adaptations and their root-associated bacterial communities. The results indicated that osmotic stress contributed to a reduction in height and increased levels of reactive oxygen species (ROS) and malondialdehyde (MDA). In response, A. sparsifolia exhibited a series of morphophysiological adjustments, including increased ratio of root to shoot biomass (R/S) and the number of root tip, enhanced vitality, high levels of peroxidase (POD), ascorbate peroxidase (APX), and glutathione (GSH), as well as osmolytes (proline, soluble protein, and soluble sugar) and modification in phytohormones (abscisic acid (ABA) and jasmonic acid (JA)). Additionally, osmotic stress resulted in alterations in the compositions and co-occurrence patterns of root-associated bacterial communities, but not α-diversity (Chao1). Specifically, the rhizosphere Actinobacteria phylum was significantly increased by osmotic stress. These shifts in root-associated bacterial communities were significantly correlated with the host's adaptation to osmotic stress. Overall, the findings revealed that osmotic stress, in addition to its impacts on plant physiology, resulted in a restructuring of root-associated microbial communities and suggested that the concomitant adjustment in plant microbiota may potentially contribute to the survival of desert plants under extreme environmental stress.

Combined effects of planting patterns and mowing time on different organs and soil stoichiometry of Cyperus esculentus in desert oasis transition zone.

There are many different planting methods for crops, however it is very important to improve the distribution ratio of elements in different organs of crops. Therefore, to understand the effect of different planting patterns on crop element balance, we selected Cyperus esculentus continuous cropping (CC) and C. esculentus - wheat rotation cropping (RC). The leaves, tubers, roots, and soil samples were taken at the mowing time (August 1st, on the 81st day after seed sowing; August 24th, on the 105th day after seed sowing; September 16th, on the 128th day after seed sowing). Results showed that CC and RC had significant effects on soil SO42- and Cl-. With the mowing time, the relative abundance of TN (total nitrogen) in tubers showed an increasing trend, the relative richness of TN in roots decreased, and the relative content of TN in leaves showed no change in the trend under the two planting modes. CC significantly increased the TN/TP (total phosphorus) of leaves, roots, and tubers. However, RC significantly increased the AN (available nitrogen)/AP (available phosphorus) of soil. The random forest analysis (RF) showed that abiotic factors contributed the most to TN/TK (total potassium) of roots, followed by TN/TK of tubers and TP/TK of roots. We found that abiotic factors had no significant impact on TP/TK of leaves and TN/TP of tubers. As expected, different planting patterns alter the plant's N (nitrogen)/P (phosphorus)/K (potassium), which in turn may modify N and P conservation strategies.

Effects of different tillage systems and mowing time on nutrient accumulation and forage nutritive value of Cyperus esculentus.

Revealing the complex relationships between management practices, crop growth, forage nutritive value and soil quality will facilitate the development of more sustainable agricultural and livestock production systems. Cyperus esculentus is known as the king of oil crops and high-quality forage. However, there is little information about the effects of different planting modes {continuous cropping (CC)/rotation cropping (RC)} and initial mowing time on the plant nutrient accumulation and forage nutritive value. Here, in a field experiment, we designed two planting patterns, C. esculentus CC and C. esculentus - wheat RC. The leaves, tubers, roots, and soil samples were collected at three mowing time (on the 78th, 101th, and 124th days after seed sowing). Results revealed that RC significantly increased the total nitrogen (TN) and potassium (TK) content of the tuber (p<0.05), while significantly decreased the TN, total phosphorus (TP), crude protein (CP), and acid detergent fiber (ADF) contents of the leaves. Under the CC pattern, the TN, TP, and TK content of roots increased significantly on the 78th days after seed sowing, and the TK content of tubers increased significantly. Under the RC pattern, the ether extract (EE) content of tubers increased significantly on the 124th days after seed sowing, while the CP and TN content of leaves decreased significantly. Correlation analysis showed that soil pH was negatively correlated with TN content in leaves, tubers, and roots. The structural equation model showed that the soil pH directly affected the plant nutrient accumulation and forage nutritive value (ß=0.68) via regulating these properties by changing soil available nutrients, anions, cations, and total nutrients. Overall, we propose that RC for C. esculentus-wheat is should not be recommended to maximize tubers and forage yield.

Alhagi sparsifolia Harbors a Different Root-Associated Mycobiome during Different Development Stages.

The mycobiome in the rhizosphere and within the roots benefits the nutrition and function of host plants. However, compared with the bacterial community, root-associated mycobiomes of desert plants and the forces that drive their assemblage are limited. Here, we investigated the mycobiomes in bulk soil, rhizosphere, and root compartments of Alhagi sparsifolia Shap., a phreatophyte species dominating in Central Asia. The internal transcribed spacer (ITS) gene phylogenetic profiles displayed significantly diverse mycobiomes across three compartments and host growth times, together explaining 31.45% of the variation in the community composition. The community structure of the perennial stage was markedly different from that of other stages (30 days to 2 years old). Along the soil-plant continuum, the α-diversity (estimated by Chao1) decreased gradually, while concomitantly increasing the community dissimilarity and the influence of edaphic factors. Specific leaf area, soil water content, and soil organic matter levels were common factors driving the composition of the three mycobiome communities. A more complex and connected network was observed in the root community compared with the other compartments. Overall, our work suggests that an age-sensitive host effect restructured the desert-plant-root-associated mycobiome, and that edaphic factors and host growth strategy may play potential roles in this process.

Phosphorous Supplementation Alleviates Drought-Induced Physio-Biochemical Damages in Calligonum mongolicum.

Calligonum mongolicum is a phreatophyte playing an important role in sand dune fixation, but little is known about its responses to drought and P fertilization. In the present study, we performed a pot experiment to investigate the effects of P fertilization under drought or well-watered conditions on multiple morpho-physio-biochemical attributes of C. mongolicum seedlings. Drought stress leads to a higher production of hydrogen peroxide (H2O2) and malondialdehyde (MDA), leading to impaired growth and metabolism. However, C. mongolicum exhibited effective drought tolerance strategies, including a higher accumulation of soluble sugars, starch, soluble protein, proline, and significantly higheractivities of peroxidase (POD) and catalase (CAT) enzymes. P fertilization increased the productivity of drought-stressed seedlings by increasing their growth, assimilative shoots relative water content, photosynthetic pigments, osmolytes accumulation, mineral nutrition, N assimilation, and reduced lipid peroxidation. Our findings suggest the presence of soil high P depletion and C. mongolicum high P requirements during the initial growth stage. Thus, P can be utilized as a fertilizer to enhance the growth and productivity of Calligonum vegetation and to reduce the fragility of the hyper-arid desert of Taklamakan in the context of future climate change.

Alhagi sparsifolia acclimatizes to saline stress by regulating its osmotic, antioxidant, and nitrogen assimilation potential.

BACKGROUND: Alhagi sparsifolia (Camelthorn) is a leguminous shrub species that dominates the Taklimakan desert's salty, hyperarid, and infertile landscapes in northwest China. Although this plant can colonize and spread in very saline soils, how it adapts to saline stress in the seedling stage remains unclear so a pot-based experiment was carried out to evaluate the effects of four different saline stress levels (0, 50, 150, and 300 mM) on the morphological and physio-biochemical responses in A. sparsifolia seedlings. RESULTS: Our results revealed that N-fixing A. sparsifolia has a variety of physio-biochemical anti-saline stress acclimations, including osmotic adjustments, enzymatic mechanisms, and the allocation of metabolic resources. Shoot-root growth and chlorophyll pigments significantly decreased under intermediate and high saline stress. Additionally, increasing levels of saline stress significantly increased Na+ but decreased K+ concentrations in roots and leaves, resulting in a decreased K+/Na+ ratio and leaves accumulated more Na + and K + ions than roots, highlighting their ability to increase cellular osmolarity, favouring water fluxes from soil to leaves. Salt-induced higher lipid peroxidation significantly triggered antioxidant enzymes, both for mass-scavenging (catalase) and cytosolic fine-regulation (superoxide dismutase and peroxidase) of H2O2. Nitrate reductase and glutamine synthetase/glutamate synthase also increased at low and intermediate saline stress levels but decreased under higher stress levels. Soluble proteins and proline rose at all salt levels, whereas soluble sugars increased only at low and medium stress. The results show that when under low-to-intermediate saline stress, seedlings invest more energy in osmotic adjustments but shift their investment towards antioxidant defense mechanisms under high levels of saline stress. CONCLUSIONS: Overall, our results suggest that A. sparsifolia seedlings tolerate low, intermediate, and high salt stress by promoting high antioxidant mechanisms, osmolytes accumulations, and the maintenance of mineral N assimilation. However, a gradual decline in growth with increasing salt levels could be attributed to the diversion of energy from growth to maintain salinity homeostasis and anti-stress oxidative mechanisms.

Foliar P-Fractions Allocation of Karelinia caspia and Tamarix ramosissima Are Driven by Soil and Groundwater Properties in a Hyper-Arid Desert Ecosystem.

The allocation patterns of foliar phosphorus (P) fractions across various vegetation types generally reflect the adaptability to P-impoverished environments. However, the allocation of foliar-P fractions within the desert herb Karelinia caspia (K. caspica) and shrub Tamarix ramosissima (T. ramosissima) in soils with different environment-P availability and the impact of soil and groundwater properties on foliar-P fractions allocation remain unclear. The foliar-P fractions (metabolites-P, nucleic acid-P, structural-P, and residual-P) of K. caspica and T. ramosissima and the properties of 0-60 cm deep soil under their canopy and groundwater were determined at four different environment-P sites. Results found that as environment-P availability decreased, both plants allocated the higher proportions of foliar-P to nucleic acid-P than to metabolites-P and structural-P. With the exception of residual-P, foliar-P fractions were markedly higher for K. caspica than T. ramosissima. Soil Olsen-P, NO3 --N, soil water content, electrical conductivity (EC), groundwater EC, and total dissolved solids (TDSs) played an important role in allocating foliar P-fractions for both K. caspica and T. ramosissima. Compared with K. caspica, the foliar-P fractions of T. ramosissima were more tightly bounded to groundwater than soil properties. Overall, these findings show how desert plants flexibility take advantage of the foliar-P in low environment-P availability and illustrate the foliar-P fractions allocation of desert plants is driven by soil and groundwater properties.

Intercropping Systems Modify Desert Plant-Associated Microbial Communities and Weaken Host Effects in a Hyper-Arid Desert.

Intercropping is an important practice in promoting plant diversity and productivity. Compared to the accumulated understanding of the legume/non-legume crop intercrops, very little is known about the effect of this practice when applied to native species on soil microbial communities in the desert ecosystem. Therefore, in the present study, bulk soil and rhizosphere microbial communities in the 2-year Alhagi sparsifolia (legume)/Karelinia caspica (non-legume) monoculture vs. intercropping systems were characterized under field conditions. Our result revealed that plant species identities caused a significant effect on microbial community composition in monocultures but not in intercropping systems. Monoculture weakened the rhizosphere effect on fungal richness. The composition of bacterial and fungal communities (ß-diversity) was significantly modified by intercropping, while bacterial richness (Chao1) was comparable between the two planting patterns. Network analysis revealed that Actinobacteria, α- and γ-proteobacteria dominated bulk soil and rhizosphere microbial co-occurrence networks in each planting pattern. Intercropping systems induced a more complex rhizosphere microbial community and a more modular and stable bulk soil microbial network. Keystone taxa prevailed in intercropping systems and were Actinobacteria-dominated. Overall, planting patterns and soil compartments, not plant identities, differentiated root-associated microbiomes. Intercropping can modify the co-occurrence patterns of bulk soil and rhizosphere microorganisms in desert ecosystems. These findings provided a potential strategy for us to manipulate desert soil microbial communities and optimize desert species allocation in vegetation sustainability.

Development of polymorphic EST-SSR markers and characterization of the autotetraploid genome of sainfoin (Onobrychis viciifolia).

BACKGROUND: Sainfoin (Onobrychis viciifolia) is a highly nutritious, tannin-containing, and tetraploid forage legume. Due to the lack of detailed transcriptomic and genomic information on this species, genetic and breeding projects for sainfoin improvement have been significantly hindered. METHODS: In this study, a total of 24,630,711 clean reads were generated from 14 different sainfoin tissues using Illumina paired-end sequencing technology and deposited in the NCBI SRA database (SRX3763386). From these clean reads, 77,764 unigene sequences were obtained and 6,752 EST-SSRs were identified using de novo assembly. A total of 2,469 primer pairs were designed, and 200 primer pairs were randomly selected to analyze the polymorphism in five sainfoin wild accessions. RESULTS: Further analysis of 40 sainfoin individuals from the five wild populations using 61 EST-SSR loci showed that the number of alleles per locus ranged from 4 to 15, and the expected heterozygosity varied from 0.55 to 0.91. Additionally, by counting the EST-SSR band number and sequencing the three or four bands in one sainfoin individual, sainfoin was confirmed to be autotetraploid. This finding provides a high level of information about this plant. DISCUSSION: Through this study, 61 EST-SSR markers were successfully developed and shown to be useful for genetic studies and investigations of population genetic structures and variabilities among different sainfoin accessions.

Full-length transcript sequencing and comparative transcriptomic analysis to evaluate the contribution of osmotic and ionic stress components towards salinity tolerance in the roots of cultivated alfalfa (Medicago sativa L.).

BACKGROUND: Alfalfa is the most extensively cultivated forage legume. Salinity is a major environmental factor that impacts on alfalfa's productivity. However, little is known about the molecular mechanisms underlying alfalfa responses to salinity, especially the relative contribution of the two important components of osmotic and ionic stress. RESULTS: In this study, we constructed the first full-length transcriptome database for alfalfa root tips under continuous NaCl and mannitol treatments for 1, 3, 6, 12, and 24 h (three biological replicates for each time points, including the control group) via PacBio Iso-Seq. This resulted in the identification of 52,787 full-length transcripts, with an average length of 2551 bp. Global transcriptional changes in the same 33 stressed samples were then analyzed via BGISEQ-500 RNA-Seq. Totals of 8861 NaCl-regulated and 8016 mannitol-regulated differentially expressed genes (DEGs) were identified. Metabolic analyses revealed that these DEGs overlapped or diverged in the cascades of molecular networks involved in signal perception, signal transduction, transcriptional regulation, and antioxidative defense. Notably, several well characterized signalling pathways, such as CDPK, MAPK, CIPK, and PYL-PP2C-SnRK2, were shown to be involved in osmotic stress, while the SOS core pathway was activated by ionic stress. Moreover, the physiological shifts of catalase and peroxidase activity, glutathione and proline content were in accordance with dynamic transcript profiles of the relevant genes, indicating that antioxidative defense system plays critical roles in response to salinity stress. CONCLUSIONS: Overall, our study provides evidence that the response to salinity stress in alfalfa includes both osmotic and ionic components. The key osmotic and ionic stress-related genes are candidates for future studies as potential targets to improve resistance to salinity stress via genetic engineering.

Multiple Regulatory Networks Are Activated during Cold Stress in Medicago sativa L.

Cultivated alfalfa (Medicago sativa L.) is one of the most important perennial legume forages in the world, and it has considerable potential as a valuable forage crop for livestock. However, the molecular mechanisms underlying alfalfa responses to cold stress are largely unknown. In this study, the transcriptome changes in alfalfa under cold stress at 4 °C for 2, 6, 24, and 48 h (three replicates for each time point) were analyzed using the high-throughput sequencing platform, BGISEQ-500, resulting in the identification of 50,809 annotated unigenes and 5283 differentially expressed genes (DEGs). Metabolic pathway enrichment analysis demonstrated that the DEGs were involved in carbohydrate metabolism, photosynthesis, plant hormone signal transduction, and the biosynthesis of amino acids. Moreover, the physiological changes of glutathione and proline content, catalase, and peroxidase activity were in accordance with dynamic transcript profiles of the relevant genes. Additionally, some transcription factors might play important roles in the alfalfa response to cold stress, as determined by the expression pattern of the related genes during 48 h of cold stress treatment. These findings provide valuable information for identifying and characterizing important components in the cold signaling network in alfalfa and enhancing the understanding of the molecular mechanisms underlying alfalfa responses to cold stress.

Transcriptome Analyses Reveal Candidate Pod Shattering-Associated Genes Involved in the Pod Ventral Sutures of Common Vetch (Vicia sativa L.).

The seed dispersion caused by pod shattering is a form of propagation used by many wild species. Loss of seeds from pod shattering is frequent in the common vetch (Vicia sativa L.), an important self-pollinating annual forage legume. However, pod shattering is one of the most important defects that limits the reproduction of the vetch in the field and the usage as a leguminous forage crop. To better understand the vetch pod shattering mechanism, we used high-throughput RNA sequencing to assess the global changes in the transcriptomes of the pod ventral sutures of shattering-susceptible and shattering-resistant vetch accessions screened from 541 vetch germplasms. A total of 1,285 significantly differentially expressed unigenes (DEGs) were detected, including 575 up-regulated unigenes and 710 down-regulated unigenes. Analyses of Gene Ontology and KEGG metabolic enrichment pathways of 1,285 DEGs indicated that 22 DEGs encoding cell wall modifications and hydrolases associated with pod shattering were highly expressed in shattering-susceptible accessions. These genes were mainly enriched in "hydrolase activity," "cytoplasm," and "carbohydrate metabolic process" systems. These cell wall modifications and hydrolases genes included ß-glucosidase and endo-polygalacturonase, which work together to break down the glycosidic bonds of pectin and cellulose, and to promote the dissolution and disappearance of the cell wall in the ventral suture of the pod and make the pod more susceptible to shattering. We demonstrated the differences in gene transcription levels between the shattering-susceptible and shattering-resistant vetch accessions for the first time and our results provided valuable information for the identifying and characterizing of pod shattering regulation networks in vetch. This information may facilitate the future identification of pod shattering-related genes and their underlying molecular mechanisms in the common vetch.

Optimizing Sample Size to Assess the Genetic Diversity in Common Vetch (Vicia sativa L.) Populations Using Start Codon Targeted (SCoT) Markers.

Common vetch (Vicia sativa subsp. sativa L.) is a self-pollinating annual forage legume with worldwide importance. Here, we investigate the optimal number of individuals that may represent the genetic diversity of a single population, using Start Codon Targeted (SCoT) markers. Two cultivated varieties and two wild accessions were evaluated using five SCoT primers, also testing different sampling sizes: 1, 2, 3, 5, 8, 10, 20, 30, 40, 50, and 60 individuals. The results showed that the number of alleles and the Polymorphism Information Content (PIC) were different among the four accessions. Cluster analysis by Unweighted Pair Group Method with Arithmetic Mean (UPGMA) and STRUCTURE placed the 240 individuals into four distinct clusters. The Expected Heterozygosity (HE) and PIC increased along with an increase in sampling size from 1 to 10 plants but did not change significantly when the sample sizes exceeded 10 individuals. At least 90% of the genetic variation in the four germplasms was represented when the sample size was 10. Finally, we concluded that 10 individuals could effectively represent the genetic diversity of one vetch population based on the SCoT markers. This study provides theoretical support for genetic diversity, cultivar identification, evolution, and marker-assisted selection breeding in common vetch.