Comprehensive Transcriptome and Metabolome Analyses Reveal Primary Molecular Regulation Pathways Involved in Peanut under Water and Nitrogen Co-Limitation.

The yield and quality of peanut (Arachis hypogaea L.), an oil crop planted worldwide, are often limited by drought stress (DS) and nitrogen (N) deficiency. To investigate the molecular mechanism by which peanut counteracts DS and N deficiency, we conducted comprehensive transcriptomic and metabolomic analyses of peanut leaves. Herein, 829 known differentially accumulated metabolites, 324 differentially expressed transcription factors, and 5294 differentially expressed genes (DEGs) were identified under different water and N conditions. The transcriptome analysis demonstrated that drought-related DEGs were predominantly expressed in "glycolysis/gluconeogenesis" and "glycerolipid metabolism", while N-deficiency-related DEGs were mainly expressed in starch and sucrose metabolism, as well as in the biosynthesis of amino acid pathways. The biosynthesis, transport, and catabolism of secondary metabolites accounted for a large proportion of the 1317 DEGs present in water and N co-limitation. Metabolomic analysis showed that the metabolic accumulation of these pathways was significantly dependent on the stress conditions. Additionally, the roles of metabolites and genes in these pathways, such as the biosynthesis of amino acids and phenylpropanoid biosynthesis under different stress conditions, were discussed. The results demonstrated that different genes, metabolic pathways, and metabolites were related to DS and N deficiency. Thus, this study elucidates the metabolic pathways and functional genes that can be used for the improvement of peanut resistance to abiotic stress.

Green manure increases peanut production by shaping the rhizosphere bacterial community and regulating soil metabolites under continuous peanut production systems.

BACKGROUND: Green manure (GM) is a crop commonly grown during fallow periods, which has been applied in agriculture as a strategy to regulate nutrient cycling, improve organic matter, and enhance soil microbial biodiversity, but to date, few studies have examined the effects of GM treatments on rhizosphere soil bacterial community and soil metabolites from continuous cropping peanut field. RESULTS: In this study, we found that the abundances of several functionally significant bacterial groups containing Actinobacteria, Acidobacteria, and genus Sphingomonas, which are associated with nitrogen cycling, were dramatically increased in GM-applied soils. Consistent with the bacterial community results, metabolomics analysis revealed a strong perturbation of nitrogen- or carbon-related metabolisms in GM-applied soils. The substantially up-regulated beneficial metabolites including sucrose, adenine, lysophosphatidylcholine (LPC), malic acid, and betaines in GM-applied soils may contribute to overcome continuous cropping obstacle. In contrast to peanut continuous cropping, planting winter wheat and oilseed rape in winter fallow period under continuous spring peanut production systems evidently improved the soil quality, concomitantly with raised peanut pod yield by 32.93% and 25.20%, in the 2020 season, respectively. CONCLUSIONS: GMs application is an effective strategy to overcome continuous cropping obstacle under continuous peanut production systems by improving nutrient cycling, soil metabolites, and rhizobacterial properties.

Comprehensive effects of salt stress and peanut cultivars on the rhizosphere bacterial community diversity of peanut.

Plant rhizosphere bacterial communities are central to plant growth and stress tolerance, which differ across cultivars and external environments. The goal of this study was to assess the comprehensive effects of salt stress and peanut cultivars on rhizosphere bacterial community diversity. In this study, we investigated the effects of salt stress on peanut morphology and pod yield and the associated rhizosphere bacterial diversity using statistical analysis and 16S rRNA gene sequencing, respectively. Statistical analysis exhibited that salt stress indeed affected peanut growth and pod yield, and various peanut cultivars showed divergences. Taxonomic analysis showed that the bacterial community predominantly consisted of phyla Actinobacteria, Proteobacteria, Chloroflexi, Acidobacteria, and Cyanobacteria in peanut rhizosphere soils. Among these bacteria, numbers of beneficial bacteria Cyanobacteria and Proteobacteria increased, especially in the salt-resistant cultivars, while that of Acidobacteria decreased after salt treatment. Nitrogen-fixing bacterium Rhizobium closely related to peanut nodulation was significantly improved in rhizosphere soils of salt-resistant cultivars after salt treatment. Metabolic function prediction showed that the percentages of reads categorized to signaling transduction and inorganic ion transport and metabolism were higher in the salt-treated soils, which may be conducive to peanut survival and salt tolerance to some extent. The study is, therefore, crucially important to develop the foundation for improving the salt tolerance of various peanut cultivars via modifying the soil bacterial community.

The synergy effect of arbuscular mycorrhizal fungi symbiosis and exogenous calcium on bacterial community composition and growth performance of peanut (Arachis hypogaea L.) in saline alkali soil.

Peanut (Arachis hypogaea. L) is an important oil seed crop. Both arbuscular mycorrhizal fungi (AMF) symbiosis and calcium (Ca2+) application can ameliorate the impact of saline soil on peanut production, and the rhizosphere bacterial communities are also closely correlated with peanut salt tolerance; however, whether AMF and Ca2+ can withstand high-salinity through or partially through modulating rhizosphere bacterial communities is unclear. Here, we used the rhizosphere bacterial DNA from saline alkali soil treated with AMF and Ca2+ alone or together to perform high-throughput sequencing of 16S rRNA genes. Taxonomic analysis revealed that AMF and Ca2+ treatment increased the abundance of Proteobacteria and Firmicutes at the phylum level. The nitrogen-fixing bacterium Sphingomonas was the dominant genus in these soils at the genus level, and the soil invertase and urease activities were also increased after AMF and Ca2+ treatment, implying that AMF and Ca2+ effectively improved the living environment of plants under salt stress. Moreover, AMF combined with Ca2+ was better than AMF or Ca2+ alone at altering the bacterial structure and improving peanut growth in saline alkali soil. Together, AMF and Ca2+ applications are conducive to peanut salt adaption by regulating the bacterial community in saline alkali soil.

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A pot experiment with Huayu 25 as experimental material was conducted, with treatments of drought and salt stresses. The effects of drought and salt stresses at the flowering stage on the plant morphology, pod yield, and soil bacterial community structure in the rhizosphere were examined. The results showed that Proteobacteria, Actinobacteria, Saccharibacteria, Chloroflexi, Cyanobacteria, and Acidobacteria were the dominant phyla in the rhizosphere soil of peanut. Compared with that under normal conditions, the relative abundance of Proteobacteria and Actinobacteria dramatically decreased, while that of Cyanobacteria evidently increased in drought-treated and salt-treated soil. Moreover, the variation of Cyanobacteria abundance caused by combined drought and salt stresses was stronger than that caused by single drought or salt stress. Functional meta-genomic profiling indicated that a series of sequences related to signaling transduction, defense mechanism and post-translational modification, protein turnover, chaperones were enriched in rhizosphere soil under stressed conditions, which might have implications for plant survival and stress tolerance. Drought and salt stress affectedpeanut growth and reduced pod yield. Results from this study would present reference on the future improvement of stress tolerance of peanuts via modifying soil microbial community.

Influence of Salt Stress on Growth of Spermosphere Bacterial Communities in Different Peanut (Arachis hypogaea L.) Cultivars.

BACKGROUND: Exposure of seeds to high salinity can cause reduced germination and poor seedling establishment. Improving the salt tolerance of peanut (Arachis hypogaea L.) seeds during germination is an important breeding goal of the peanut industry. Bacterial communities in the spermosphere soils may be of special importance to seed germination under salt stress, whereas extant results in oilseed crop peanut are scarce. METHODS: Here, bacterial communities colonizing peanut seeds with salt stress were characterized using 16S rRNA gene sequencing. RESULTS: Peanut spermosphere was composed of four dominant genera: Bacillus, Massilia, Pseudarthrobacter, and Sphingomonas. Comparisons of bacterial community structure revealed that the beneficial bacteria (Bacillus), which can produce specific phosphatases to sequentially mineralize organic phosphorus into inorganic phosphorus, occurred in relatively higher abundance in salt-treated spermosphere soils. Further soil enzyme activity assays showed that phosphatase activity increased in salt-treated spermosphere soils, which may be associated with the shift of Bacillus. CONCLUSION: This study will form the foundation for future improvement of salt tolerance of peanuts at the seed germination stage via modification of the soil microbes.

Influence of salt stress on the rhizosphere soil bacterial community structure and growth performance of groundnut (Arachis hypogaea L.).

Soil salinity is regarded as severe environmental stress that can change the composition of rhizosphere soil bacterial community and import a plethora of harms to crop plants. However, relatively little is known about the relationship between salt stress and root microbial communities in groundnuts. The goal of this study was to assess the effect of salt stress on groundnut growth performance and rhizosphere microbial community structure. Statistical analysis exhibited that salt stress indeed affected groundnut growth and pod yield. Further taxonomic analysis showed that the bacterial community predominantly consisted of phyla Proteobacteria, Actinobacteria, Saccharibacteria, Chloroflexi, Acidobacteria, and Cyanobacteria. Among these bacteria, numbers of Cyanobacteria and Acidobacteria mainly increased, while that of Actinobacteria and Chloroflexi decreased after salt treatment via taxonomic and qPCR analysis. Moreover, Sphingomonas and Microcoleus as the predominant genera in salt-treated rhizosphere soils might enhance salt tolerance as plant growth-promoting rhizobacteria. Metagenomic profiling showed that series of sequences related to signaling transduction, posttranslational modification, and chaperones were enriched in the salt-treated soils, which may have implications for plant survival and salt tolerance. These data will help us better understand the symbiotic relationship between the dominant microbial community and groundnuts and form the foundation for further improvement of salt tolerance of groundnuts via modification of soil microbial community.

Effect of Drought Stress and Developmental Stages on Microbial Community Structure and Diversity in Peanut Rhizosphere Soil.

BACKGROUND: Peanut (Arachis hypogaea L.), an important oilseed and food legume, is widely cultivated in the semi-arid tropics. Drought is the major stress in this region which limits productivity. Microbial communities in the rhizosphere are of special importance to stress tolerance. However, relatively little is known about the relationship between drought and microbial communities in peanuts. METHOD: In this study, deep sequencing of the V3-V4 region of the 16S rRNA gene was performed to characterize the microbial community structure of drought-treated and untreated peanuts. RESULTS: Taxonomic analysis showed that Actinobacteria, Proteobacteria, Saccharibacteria, Chloroflexi, Acidobacteria and Cyanobacteria were the dominant phyla in the peanut rhizosphere. Comparisons of microbial community structure of peanuts revealed that the relative abundance of Actinobacteria and Acidobacteria dramatically increased in the seedling and podding stages in drought-treated soil, while that of Cyanobacteria and Gemmatimonadetes increased in the flowering stage in drought-treated rhizospheres. Metagenomic profiling indicated that sequences related to metabolism, signaling transduction, defense mechanism and basic vital activity were enriched in the drought-treated rhizosphere, which may have implications for plant survival and drought tolerance. CONCLUSION: This microbial communities study will form the foundation for future improvement of drought tolerance of peanuts via modification of the soil microbes.

[Diversity of microbial community structure in the spermosphere of saline-alkali soil in shandong area].

Objective: Three soil types in different salt contents were taken as the experiment objectives. We evaluated the effect of various saline alkali soil types on diversity of bacterial community structure in spermosphere soil during water absorption and germination of peanut seeds. Methods: The V3-V4 region of 16S ribosomal RNA genes was amplified using PCR, and the PCR products were then analyzed using Illumina high-throughput sequencing technology. Results: (1) The diversity of soil bacterial community in saline alkali soil was higher than that in non-saline alkali soil. Especially, the highest diversity was in spermosphere soil from Qingtuo. (2) The microflora structures in different soils were distinct at the class level. Soil bacteria in four samples were classified into six classes, including Proteobacteria, Actinobacteria, Actinobacteria, Bacteroidetes, Acidobacteria and Firmicutes. Proteobacteria and Actinobacteria groups were dominant in colonies. The analysis of whole samples colony structure showed that the difference of type and abundance at phylum and genus level during different adsorption time was most significant (P<0.05). (3) The analysis of beta diversity and phylogenetic distances of constructed phylogenetic trees revealed that the sequenced clones fell into two major groups within the domain bacteria. Conclusion: The diversity of bacteria community compositions in the high salt content soil was higher. There were obvious differences in microbial community structure of different soil types at class level, primarily in the Proteobacteria and Actinobacteria. The type and abundance of microbial colonies at both phylum and genus levels were affected by the seed germination time. However, there was no influence on the genetic distance between the samples from the same soil type.

[Effects of water stress and nitrogen fertilization on peanut root morphological development and leaf physiological activities].

Taking 'Huayu 22' peanut as test material, effect of soil water content and nitrogen fertilization on the leaf physiological activities and root morphological characteristics of peanut plants were analyzed. Two levels of soil water condition were: (1) well-watered condition and (2) moderate water stress, and three levels of nitrogen were: (1) none nitrogen (N0), (2) moderate nitrogen (N1, 90 kg · hm(-2)) and (3) high nitrogen (N2, 180 kg · hm(-2)). The results showed that N1 significantly increased the peanut yield under two water conditions, but showed no significant effect on harvest index compared with N0. Under water stress condition, N1 had no significant effects on total root biomass and total root length, but the total root surface area was remarkably increased. The nitrogen fertilization significantly increased the root length and root surface area in 20-40 cm soil layer, and N2 significantly increased the root biomass and root surface area in the soil layer below 40 cm. The application of nitrogen remarkably increased CAT and POD activities in leaf, while MDA content was decreased with the increase of nitrogen level. Under well-watered condition, the root biomass, root length and root surface area in the soil layer below 40 cm and total root surface area were significantly reduced by nitrogen application, however, only N1 could increase leaf protective enzyme activities. Correlation analysis showed that the root length in 20-40 cm soil layer and SOD, CAT, POD activities in leaf were highly significantly related with peanut yield.

[Effects of drought stress on the root growth and development and physiological characteristics of peanut].

Taking two peanut varieties Huayu 17 and Tangke 8 as test objects, a soil column culture experiment was conducted in a rainproof tank to study the peanut root morphological development and physiological characteristics at late growth stages under moderate drought and well-watered conditions. Tanke 8 had more developed root system and higher yield and drought coefficient, while Huayu 17 had poorer root adaptability to drought stress. For the two varieties, their root length density and root biomass were mainly distributed in 0-40 cm soil layer, whereas their root traits differed in the same soil layer. The total root length, total root surface area, and total root volume of Huayu 17 at each growth stage were smaller under drought stress than under well-balanced water treatment, while these root characteristics of Tangke 8 under drought stress only decreased at flowering-pegging stage. Drought stress increased the root biomass, surface area, and volume of the two varieties in 20-40 cm soil layer, but decreased these root traits in the soil layers below 40 cm. Under drought stress, the root activity of the two varieties in the soil layers below 40 cm at pod filling stage decreased, and the decrement was larger for Huayu 17. The differences in the root system development and physiological characteristics of the two varieties at late growth stages under drought stress suggested that the root system of the two varieties had different water absorption and utilization under drought stress.

[Indices selection and comprehensive evaluation of salinity tolerance for peanut varieties].

A total of two hundred peanut varieties (lines) were exposed to different salt concentrations under pot cultivation, to evaluate salinity tolerance by indices such as emergence, morphology and biomass accumulation from emergence to seedling stage. The results showed that, as the salinity concentration increased, the emergence time was prolonged, plant morphology establishment was inhibited seriously, and biomass accumulation was reduced. The optimal concentration for evaluating salinity tolerance was 0.30%-0.45%. Ten indices were contributed to the mean membership function value by the membership function analysis. According to the correlation coefficient between indices and the mean membership function value, plant fresh mass, shoot fresh mass, root fresh mass, root dry mass, plant height and stem height could be the first selected indices for evaluating salinity tolerance of peanut plant. Plant dry mass, shoot dry mass, taproot length and emergence speed could be the second selected indices to comprehensively evaluate salinity tolerance of peanut plant. The 200 varieties were divided into 4 groups at different salinity concentrations, i. e. high salinity tolerance, salinity tolerance, salinity sensitivity, and high salinity sensitivity. Number of salinity tolerant varieties was decreased with increasing salinity concentration while the salinity sensitive one was increased. Salinity tolerance of some varieties showed the similarity (tolerant or sensitive) under different salinity stresses. Some varieties showed different tolerance under different salinity stresses, i. e. tolerance at low salinity concentration while sensitivity at high salinity concentration.