Continuous planting of euhalophyte Suaeda salsa enhances microbial diversity and multifunctionality of saline soil.

Halophyte-based remediation emerges as a novel strategy for ameliorating saline soils, offering a sustainable alternative to conventional leaching methods. While bioremediation is recognized for its ability to energize soil fertility and structure, the complex interplays among plant traits, soil functions, and soil microbial diversity remain greatly unknown. Here, we conducted a 5-year field experiment involving the continuous cultivation of the annual halophyte Suaeda salsa in saline soils to explore soil microbial diversity and their relationships with plant traits and soil functions. Our findings demonstrate that a decline in soil salinity corresponded with increases in the biomass and seed yield of S. salsa, which sustained a consistent seed oil content of approximately 22% across various salinity levels. Significantly, prolonged cultivation of halophytes substantially augmented soil microbial diversity, particularly from the third year of cultivation. Moreover, we identified positive associations between soil multifunctionality, seed yield, and taxonomic richness within a pivotal microbial network module. Soils enriched with taxa from this module showed enhanced multifunctionality and greater seed yields, correlating with the presence of functional genes implicated in nitrogen fixation and nitrification. Genomic analysis suggests that these taxa have elevated gene copy numbers of crucial functional genes related to nutrient cycling. Overall, our study emphasizes that the continuous cultivation of S. salsa enhances soil microbial diversity and recovers soil multifunctionality, expanding the understanding of plant-soil-microbe feedback in bioremediation.IMPORTANCEThe restoration of saline soils utilizing euhalophytes offers a viable alternative to conventional irrigation techniques for salt abatement and soil quality enhancement. The ongoing cultivation of the annual Suaeda salsa and its associated plant traits, soil microbial diversity, and functionalities are, however, largely underexplored. Our investigation sheds light on these dynamics, revealing that cultivation of S. salsa sustains robust plant productivity while fostering soil microbial diversity and multifunctionality. Notably, the links between enhanced soil multifunctionality, increased seed yield, and network-dependent taxa were found, emphasizing the importance of key microbial taxa linked with functional genes vital to nitrogen fixation and nitrification. These findings introduce a novel understanding of the role of soil microbes in bioremediation and advance our knowledge of the ecological processes that are vital for the rehabilitation of saline environments.

Utilization of halophytes in saline agriculture and restoration of contaminated salinized soils from genes to ecosystem: Suaeda salsa as an example.

Halophytes can be used to screen genes for breeding salt-tolerant crops and are of great value in the restoration of salinized or contaminated soils. However, the potential of halophytes in improving saline soils remains limited. In this paper, based on the latest research progress, we use Suaeda salsa L. as an example to evaluate the value of halophytes in developing saline agriculture including: 1) some defined salt-resistance genes and high-affinity nitrate transporter genes in the species for breeding salt-tolerance and nitrogen efficiency crops; 2) the value of S. salsa and microorganisms from S. salsa in remediation of heavy metal contaminated and organic polluted saline soils; and 3) the capacity to remove salts from soils and the application of the species. In conclusion, S. salsa has high value as a candidate to explore the theoretical base and practical application for utilizing halophytes to improve salinized soils from genes to ecosystem.

Soil pH is equally important as salinity in shaping bacterial communities in saline soils under halophytic vegetation.

While saline soils account for 6.5% of the total land area globally, it comprises about 70% of the area in northwestern China. Microbiota in these saline soils are particularly important because they are critical to maintaining ecosystem services. However, little is known about the microbial diversity and community composition in saline soils. To investigate the distribution patterns and edaphic determinants of bacterial communities in saline soils, we collected soil samples across the hypersaline Ebinur Lake shoreline in northwestern China and assessed soil bacterial communities using bar-coded pyrosequencing. Bacterial communities were diverse, and the dominant phyla (>5% of all sequences) across all soil samples were Gammaproteobacteria, Actinobacteria, Firmicutes, Alphaproteobacteria, Bacteroidetes and Betaproteobacteria. These dominant phyla made a significant (P < 0.05) contribution to community structure variations between soils. Halomonas, Smithella, Pseudomonas and Comamonas were the indicator taxa across the salinity gradient. Bacterial community composition showed significant (P < 0.05) correlations with salt content and soil pH. Indeed, bacterial phylotype richness and phylogenetic diversity were also higher in soils with middle-level salt rates, and were significantly (P < 0.05) correlated with salt content and soil pH. Overall, our results show that both salinity and pH are the determinants of bacterial communities in saline soils in northwest China.

High lithium tolerance of Apocynum venetum seeds during germination.

Identification and use of lithium (Li) accumulator plants is a promising strategy to remediate Li-contaminated soil. Apocynum venetum is reported as a Li accumulator. However, its tolerance to Li salt during germination is still unknown. The primary aim of this study was to investigate the effects of two Li salts on seed germination of A. venetum. At the same concentrations, germination percentages in LiCl solution were higher than that in Li2CO3 solution. At 25 °C, seeds germinated to 4-90% at 0-400 mmol L-1 LiCl and 3-91% at 0-150 mmol L-1 Li2CO3. Low concentration (0-50 mmol L-1) of LiCl did not significantly affect germination percentage. The simulated critical value (when germination percentage is 50%) in LiCl solution is 196 mmol L-1, and 36 mmol L-1 for Li2CO3. Activity of α-amylase, contents of MDA, soluble sugar, and proline were dramatically affected by Li salts, especially at medium and late germination stages. When compared with control, α-amylase activity of seeds under 25 mmol L-1 LiCl and 10 mmol L-1 Li2CO3 did not show significant difference. Germination percentage and index, radicle length, and physiological parameters indicate A. venetum seeds are highly tolerant to Li salts during germination, especially LiCl.

Transcriptome assembly in Suaeda aralocaspica to reveal the distinct temporal gene/miRNA alterations between the dimorphic seeds during germination.

BACKGROUND: Dimorphic seeds from Suaeda aralocaspica exhibit different germination behaviors that are thought to be a bet-hedging strategy advantageous in harsh and unpredictable environments. To understand the molecular mechanisms of Suaeda aralocaspica dimorphic seed germination, we applied RNA sequencing and small RNA sequencing for samples collected at three germination stages. RESULTS: A total of 79,414 transcripts were assembled using Trinity, of which 57.67% were functionally annotated. KEGG enrichment unveiled that photosynthesis and flavonol biosynthesis pathways were activated earlier in brown seed compared with black seed. Gene expression analysis revealed that nine candidate unigenes in gibberellic acid and abscisic acid signal transduction and 23 unigenes in circadian rhythm-plant pathway showed distinct expression profiles to promote dimorphic seed germination. 194 conserved miRNAs comprising 40 families and 21 novel miRNAs belonging to 20 families in Suaeda aralocaspica were identified using miRDeep-P and Mfold. The expression of miRNAs in black seed was suppressed at imbibition stage. Among the identified miRNAs, 59 conserved and 13 novel miRNAs differentially expressed during seed germination. Of which, 43 conserved and nine novel miRNAs showed distinct expression patterns between black and brown seed. Using TAPIR, 208 unigenes were predicted as putative targets of 35 conserved miRNA families and 17 novel miRNA families. Among functionally annotated targets, genes participated in transcription regulation constituted the dominant category, followed by genes involved in signaling and stress response. Seven of the predicted targets were validated using 5' rapid amplification of cDNA ends or real-time quantitative reverse transcription-PCR. CONCLUSIONS: Our results indicate that specific genes and miRNAs are regulated differently between black and brown seed during germination, which may contribute to the different germination behaviors of Suaeda aralocaspica dimorphic seeds in unpredictable variable environments. Our results lay a solid foundation for further studying the roles of candidate genes and miRNAs in Suaeda aralocaspica dimorphic seed germination.

Germination of dimorphic seeds of Suaeda aralocaspica in response to light and salinity conditions during and after cold stratification.

Cold stratification is a requirement for seed dormancy breaking in many species, and thus it is one of the important factors for the regulation of timing of germination. However, few studies have examined the influence of various environmental conditions during cold stratification on subsequent germination, and no study has compared such effects on the performance of dormant versus non-dormant seeds. Seeds of halophytes in the cold desert might experience different light and salinity conditions during and after cold stratification. As such, dimorphic seeds (non-dormant brown seeds and black seeds with non-deep physiological dormancy) of Suaeda aralocaspica were cold stratified under different light (12 h light-12 h darkness photoperiod or continuous darkness) or salinity (0, 200 or 1,000 mmol L-1 NaCl) conditions for 20 or 40 days. Then stratified seeds were incubated under different light or salinity conditions at daily (12/12 h) temperature regime of 10:25 °C for 20 days. For brown seeds, cold stratification was also part of the germination period. In contrast, almost no black seeds germinated during cold stratification. The longer the cold stratification, the better the subsequent germination of black seeds, regardless of light or salinity conditions. Light did not influence germination of brown seeds. Germination of cold-stratified black seeds was inhibited by darkness, especially when they were stratified in darkness. With an increase in salinity at the stage of cold stratification or germination, germination percentages of both seed morphs decreased. Combinational pre-treatments of cold stratification and salinity did not increase salt tolerance of dimorphic seeds in germination phase. Thus, light and salinity conditions during cold stratification partly interact with these conditions during germination stage and differentially affect germination of dimorphic seeds of S. aralocaspica.

Effect of halotolerant rhizobacteria isolated from halophytes on the growth of sugar beet (Beta vulgaris L.) under salt stress.

Utilization of rhizobacteria that have associated with plant roots in harsh environments could be a feasible strategy to deal with limits to agricultural production caused by soil salinity. Halophytes occur naturally in high-salt environments, and their roots may be associated with promising microbial candidates for promoting growth and salt tolerance in crops. This study aimed to isolate efficient halotolerant plant-growth-promoting rhizobacterial strains from halophytes and evaluate their activity and effects on sugar beet (Beta vulgaris L.) growth under salinity stress. A total of 23 isolates were initially screened for their ability to secrete 1-aminocyclopropane-1-carboxylate deaminase (ACD) as well as other plant-growth-promoting characteristics and subsequently identified by sequencing the 16S rRNA gene. Three isolates, identified as Micrococcus yunnanensis, Planococcus rifietoensis and Variovorax paradoxus, enhanced salt stress tolerance remarkably in sugar beet, resulting in greater seed germination and plant biomass, higher photosynthetic capacity and lower stress-induced ethylene production at different NaCl concentrations (50-125 mM). These results demonstrate that salinity-adapted, ACD-producing bacteria isolated from halophytes could promote sugar beet growth under saline stress conditions.

Isolation of Endophytic Plant Growth-Promoting Bacteria Associated with the Halophyte Salicornia europaea and Evaluation of their Promoting Activity Under Salt Stress.

Several reports have highlighted that many plant growth-promoting endophytic bacteria (PGPE) can assist their host plants in coping with various biotic and abiotic stresses. However, information about the PGPE colonizing in the halophytes is still scarce. This study was designed to isolate and characterize PGPE from salt-accumulating halophyte Salicornia europaea grown under extreme salinity and to evaluate in vitro the bacterial mechanisms related to plant growth promotion. A total of 105 isolates were obtained from the surface-sterilized roots, stems, and assimilation twigs of S. europaea. Thirty-two isolates were initially selected for their ability to produce 1-aminocyclopropane-1-carboxylate deaminase as well as other properties such as production of indole-3-acetic acid and phosphate-solubilizing activities. The 16S rRNA gene-sequencing analysis revealed that these isolates belong to 13 different genera and 19 bacterial species. For these 32 strains, seed germination and seedling growth in axenically grown S. europaea seedlings at different NaCl concentrations (50-500 mM) were quantified. Five isolates possessing significant stimulation of the host plant growth were obtained. The five isolates were identified as Bacillus endophyticus, Bacillus tequilensis, Planococcus rifietoensis, Variovorax paradoxus, and Arthrobacter agilis. All the five strains could colonize and can be reisolated from the host plant interior tissues. These results demonstrate that habitat-adapted PGPE isolated from halophyte could enhance plant growth under saline stress conditions.

High-Throughput Sequencing Analysis of the Endophytic Bacterial Diversity and Dynamics in Roots of the Halophyte Salicornia europaea.

Endophytic bacterial communities of halophyte Salicornia europaea roots were analyzed by 16S rRNA gene pyrosequencing. A total of 20,151 partial 16S rRNA gene sequences were obtained. These sequences revealed huge amounts of operational taxonomic units (OTUs), that is, 747-1405 OTUs in a root sample, at 3 % cut-off level. Root endophytes mainly comprised four phyla, among which Proteobacteria was the most represented, followed by Bacteroidetes, Actinobacteria, and Firmicutes. Gammaproteobacteria was the most abundant class of Proteobacteria, followed by Betaproteobacteria and Alphaproteobacteria. Genera Pantoea, Halomonas, Azomonas, Serpens, and Pseudomonas were shared by all growth periods. A marked difference in endophytic bacterial communities was evident in roots from different host life-history stages. Gammaproteobacteria increased during the five periods, while Betaproteobacteria decreased. The richest endophytic bacteria diversity was detected in the seedling stage. Endophytic bacteria diversity was reduced during the flowering stage and fruiting stage. The five libraries contained 2321 different OTUs with 41 OTUs in common. As a whole, this study first surveys communities of endophytic bacteria by tracing crucial stages in the process of halophyte growth using high-throughput sequencing methods.

Illumina-based analysis of bacterial diversity related to halophytes Salicornia europaea and Sueada aralocaspica.

We used Illumina-based 16S rRNA V3 amplicon pyrosequencing to investigate the community structure of soil bacteria from the rhizosphere surrounding Salicornia europaea, and endophytic bacteria living in Salicornia europaea plants and Sueada aralocaspica seeds growing at the Fukang Desert Ecosystem Observation and Experimental Station (FDEOES) in Xinjiang Province, China, using an Illumina genome analyzer. A total of 89.23 M effective sequences of the 16S rRNA gene V3 region were obtained from the two halophyte species. These sequences revealed a number of operational taxonomic units (OTUs) in the halophytes. There were between 22-2,206 OTUs in the halophyte plant sample, at the 3% cutoff level, and a sequencing depth of 30,000 sequences. We identified 25 different phyla, 39 classes and 141 genera from the resulting 134,435 sequences. The most dominant phylum in all the samples was Proteobacteria (41.61%-99.26%; average, 43.30%). The other large phyla were Firmicutes (0%- 7.19%; average, 1.15%), Bacteroidetes (0%-1.64%; average, 0.44%) and Actinobacteria (0%-0.46%; average, 0.24%). This result suggested that the diversity of bacteria is abundant in the rhizosphere soil, while the diversity of bacteria was poor within Salicornia europaea plant samples. To the extent of our knowledge, this study is the first to characterize and compare the endophytic bacteria found within different halophytic plant species roots using PCR-based Illumina pyrosequencing method.

Actinotalea suaedae sp. nov., isolated from the halophyte Suaeda physophora in Xinjiang, Northwest China.

A Gram-stain-positive, aerobic, non-motile, coryneform bacterium, designated strain EGI 60002(T), was isolated from the halophyte Suaeda physophora. Cells were coryneform shaped and polymorphic. Phylogenetic analysis based on 16S rRNA gene sequences showed that the new isolate was closely related to Actinotalea ferrariae CF5-4(T) (95.8 % gene sequence similarity). The peptidoglycan type of strain EGI 60002(T) was A4ß, containing L-Orn-D-Ser-D-Asp. The cell-wall sugars were mannose, ribose, rhamnose and glucose. The major fatty acids (>5 %) of strain EGI 60002(T) were iso-C14:0, iso-C15:0, anteiso-C15:1 A and anteiso-C15:0. The predominant respiratory quinone was MK-10(H4). The major polar lipids were diphosphatidylglycerol (DPG), one unidentified phosphoglycolipid (PGL) and one unidentified phospholipid (PL1). The genomic DNA G+C content was 72.3 mol%. On the basis of morphological, physiological, chemotaxonomic data, and phylogenetic analysis, strain EGI 60002(T) should be classified as a novel species within the genus Actinotalea, for which the name Actinotalea suaedae sp. nov. is proposed. The type strain is EGI 60002T (=JCM 19624(T) = KACC 17839(T) = KCTC 29256(T)).

Growing season carbon dioxide exchange in flooded non-mulching and non-flooded mulching cotton.

There is much interest in the role that agricultural practices might play in sequestering carbon to help offset rising atmospheric CO2 concentrations. However, limited information exists regarding the potential for increased carbon sequestration of different management strategies. The objective of this study was to quantify and contrast carbon dioxide exchange in traditional non-mulching with flooding irrigation (TF) and plastic film mulching with drip irrigation (PM) cotton (Gossypium hirsutum L.) fields in northwest China. Net primary productivity (NPP), soil heterotrophic respiration (R(h)) and net ecosystem productivity (NEP) were measured during the growing seasons in 2009 and 2010. As compared with TF, PM significantly increased the aboveground and belowground biomass and the NPP (340 g C m⁻² season⁻¹) of cotton, and decreased the R(h) (89 g C m⁻² season⁻¹) (p<0.05). In a growing season, PM had a higher carbon sequestration in terms of NEP of ∼ 429 g C m⁻² season⁻¹ than the TF. These results demonstrate that conversion of this type of land use to mulching practices is an effective way to increase carbon sequestration in the short term in cotton systems of arid areas.

Suppression of Fusarium oxysporum and induced resistance of plants involved in the biocontrol of Cucumber Fusarium Wilt by Streptomyces bikiniensis HD-087.

Cucumber Fusarium Wilt, caused by Fusarium oxysporum f. sp. cucumerinum, which usually leads to severe economic damage, is a common destructive disease worldwide. To date, no effective method has yet been found to counteract this disease. A fungal isolate, designated HD-087, which was identified as Streptomyces bikiniensis using physiological-biochemical identification and 16S rRNA sequence analysis, is shown to possess distinctive inhibitory activity against F. oxysporum. The fermentation broth of HD-087 leads to certain abnormalities in pathogen hyphae. It peroxidizes cell membrane lipids, which leads to membrane destruction along with cytoplasm leakage. This broth also restrains germination of the conidia. The activities of the enzymes peroxidase, phenylalanine ammonia-lyase, and ß-1,3-glucanase in cucumber leaves were dramatically increased after treated with fermentation broth of HD-087. The levels of chlorophyll and soluble sugars were also found to be increased, with the relative conductivity of leaves being reduced. In short, the metabolites of strain HD-087 can effectively suppress F. oxysporum and trigger induced resistance in cucumber.

[Effects of drip irrigation with plastic mulching on the net primary productivity, soil heterotrophic respiration, and net CO2 exchange flux of cotton field ecosystem in Xinjiang, Northwest China].

In April-October, 2009, a field experiment was conducted to study the effects of drip irrigation with plastic mulching (MD) on the net primary productivity (NPP), soil heterotrophic respiration (Rh) , and net CO2 exchange flux (NEF(CO2)) of cotton field ecosystem in Xinjiang, taking the traditional flood irrigation with no mulching (NF) as the control. With the increasing time, the NPP, Rh, and NEF(CO2) in treatments MD and NF all presented a trend of increasing first and decreased then. As compared with NF, MD increased the aboveground and belowground biomass and the NPP of cotton, and decreased the Rh. Over the whole growth period, the Rh in treatment MD (214 g C x m(-2)) was smaller than that in treatment NF (317 g C x m(-2)), but the NEF(CO2) in treatment MD (1030 g C x m(-2)) was higher than that in treatment NF (649 g C x m(-2)). Treatment MD could fix the atmospheric CO2 approximately 479 g C x m(-2) higher than treatment NF. Drip irrigation with plastic mulching could promote crop productivity while decreasing soil CO2 emission, being an important agricultural measure for the carbon sequestration and emission reduction of cropland ecosystems in arid area.