PART 2 Regulatory Relationship Between Quality Variation And Environment Of Cistanche Deserticola in Three Ecotypes Based On Soil Microbiome Analysis
Mar 03, 2022
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correlations amongst key microbial community abundance, PhGs content, and ecological factors. The redundant analysis of the core, biomarker microbiome abundance, PhGs content, and ecological factors was performed at the order level, and reanalysis was performed on the basis of efects. The adjusted interpretation of variance was 82.70% (Table S7). The Sphingomonadales explained 45.7% of PhGs content (p = 0.002). The Pseudonocardiales explained 22.4% of PhGs content (p = 0.01). The 2′-acetylacteosid was significantly positively correlated with Pseudonocardiales and Oceanospirillalesand negatively correlated with Sphingomonadales (Fig. 5a). The echinacoside was significantly positive with Sphingomonadales.
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Correlation analysis was conducted for biomarker abundance, PhG contents (Fig. S3 and Table S5), and ecological factors (Table S6). The results of the correlation networks (Fig. 5b) revealed that 2′-acetylacteosid was significantly positively correlated with Oceanospirillales, Rhizobiales, and Thiotrichales, whilst negatively correlated with Pseudonocardiales and Nitrosomonadales in all soil samples. The heatmap (Fig. 5c–e) revealed that tableside Awas negatively correlated with Average annual water vapor pressure in saline-alkali correlations amongst key microbial community abundance, PhGs content, and ecological factors. The redundant analysis of the core, biomarker microbiome abundance, PhGs content, and ecological factors was performed at the order level, and reanalysis was performed on the basis of efects. The adjusted interpretation of variance was 82.70% (Table S7). The Sphingomonadales explained 45.7% of PhGs content (p = 0.002). The Pseudonocardiales explained 22.4% of PhGs content (p = 0.01). The 2′-acetylacteosid was significantly positively correlated with Pseudonocardiales and Oceanospirillalesand negatively correlated with Sphingomonadales (Fig. 5a). The echinacoside was significantly positive with Sphingomonadales.
Correlation analysis was conducted for biomarker abun I land. Meanwhile, cistanoside Awas significantly positively correlated with Micrococcales and negatively correlated with Rhizobiales in saline-alkali land (Fig. 5c). In grassland,2′-acetylacteosidwas negatively correlated with average annual temperature (Fig. 5d). In sandy land, tubuloside A was negatively correlated with Nitrosomonadales (Fig. 5e).
the predictive function of the bacterial microbiome in the three ecotypes of C. deserticola. The functional profiles of the bacterial microbiome were predicted based on the 16S rRNA gene copy number of deciphered bacterial taxa using Tax4Fun according to the KEGG Ortholog groups (KOs). The results of functional prediction (Fig. 6) demonstrated that the functional metabolisms of soil microbiomes in the three ecotypes of C. deserticola were identical. Amongst the metabolisms, carbohydrate, amino acid, co-factors, and vitamin and energy metabolisms were abundant. Membrane transport and signal transduction were also abundant in environmental information processing.
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Discussion
Our previous work demonstrated that psbA-turn sequence and 2′-acetylacteosid can be used as molecular and chemical markers to distinguish C. desertica from Xinjiang and Inner Mongolia27. With field investigations, we found that C. desertica inhabits mainly three types of habitats, including saline-alkali land by EbinurLake, sandy land around Alexa League, and intermediate desert grasslands. The metabolic profiles of three ecotype C. desertica also showed that 2′-acetylacteosid can be used as a chemical marker to distinguish the three ecotypes1. We discussed the variation of C. deserticola quality and its formation mechanism from the dimensions of heredity, metabolism, and climatic factors. Therefore, from a micro perspective, the correlation network analysis
of microbiome abundance, PhGs contents, and ecological factors were conducted to elucidate the feature of soil microbial community of the three ecotypes of C. deserticola and their relationship with the quality variation.
water. Moreover, Bacillalescan produces rich metabolic products to synthesize a variety of organic acids, enzymes, physiological activities, and other substances, as well as a variety of other nutrients that can be easily utilised33. The common environmental characteristics of the three ecotypes of C. deserticola are drought and soil desertification. The soil around Ebinur Lake is also accompanied by saline-alkali stress. This may be the reason that the core soil microbial communities of three ecotypes have certain characteristics of drought, salt tolerance, alkali resistance, and stress resistance.
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Microbial factors affecting the variation of PhGs in three ecotypes of C. deserticola. Fig. S3 shows a box diagram of the PhG content of the three ecotypes of C. deserticola. The figure reveals that the 2′-acetylacteosid content is higher in saline-alkali land than in grassland and sandy land, which is consistent with the previous results1. Redundant analysis and association network results show that Oceanospirillales is significantly positively correlated with 2′-acetylacteosid. Oceanospirillales, the highest biomarker in terms of LDA score in saline-alkali land, are specifically enriched. These bacteria are metabolically and morphologically diverse, some of which can grow in the presence of oxygen whilst others require an anaerobic environment34. Oceanospirillales are an order of Proteobacteria comprising two families. Marine spirillum is often an endosymbiont of bone-eating worms (Osedax)35. Most Oceanospirillales prefer or require high salt concentrations to grow. Despite their growth in diverse niches, Oceanospirillales derive energy from the breakdown of various organic products. Therefore, the high salinity and alkalinity are the main reasons for the enrichment of Oceanospirillales in saline-alkali land soil. This result strongly suggests that the highest content of 2′-acetylacteosid in saline-alkali is related to the enrichment of Oceanospirillales. However, the regulatory relationship between Oceanospirillales and 2′-acetylacteosid is still blank, and further research is needed.
The overall contents of the seven PhGs are the highest in the grassland, amongst which echinacoside is the dominant PhGs. Echinacoside is significantly positively correlated with Sphingomonadales, which is a sequence within the alpha-proteus and constitutes the family of Erythrobacteraceae and Sphingomonadaceae. Both families are common in nature, especially in soils, oceans, and freshwater36. Sphingomonadales has a wide range of metabolic capacity for aromatic compounds, and some strains can synthesize valuable extracellular biopolymers37. All previously known members of the class Sphingomonas are aerobic and chemically organic. The only exception is the facultative anaerobic ethanol fermenter, which is used to produce fermented beverage pulp. Certain species of the genus Rhodobacter, porphyrin, and Staphylococcus aureus, as well as certain species of the genus Sphingomonas, have chlorophyll a and are therefore optional photo-organotroph (energy generated via photosynthesis)38. The best quality of C. deserticola in grassland may be due to the rich microbial community diversity and metabolic-related functions of the biomarker (Sphingomonadales). This finding provides new insight into the study on the quality variation of C. deserticola in diferent ecotypes.
Prediction metabolic function profiles of soil microbiomes of the three ecotypes C. deserticola. The metabolic function profiles (Fig. 6 and File S1) of soil microbiomes of C. deserticola in the three ecotypes were demonstrated for the frst time in this study. In terms of metabolism, carbohydrate metabolism (starch and sucrose metabolism,ko00500; amino sugar and nucleotide sugar metabolism, ko00520) and amino acid metabolism (arginine and proline metabolism, ko00330; glycine, serine, and threonine metabolism, ko00260) are highly enriched in microbiomes. Carbohydrate metabolism is responsible for the formation, breakdown, and conversion of carbohydrates in the body. Carbohydrates are the basis of many important metabolic pathways39. Carbohydrates such as glucose are part of multiple metabolic pathways across species. Carbohydrates are synthesized by plants from the atmosphere through photosynthesis and can be used as substrates for cellular respiration40. Plants and microorganisms absorb ammonia, ammonium salt, nitrite, nitrate, and other inorganic nitrogen from the environment to synthesize proteins and nitrogen-containing substances. Some microbes can convert N2 from air into ammonia nitrogen to synthesize amino acids41. The metabolic function of soil microbiomes was enriched in the primary metabolism. This enrichment suggests that the microbiomes can provide nutrition to plants and promote their growth under drought and other stresses.
Metabolic function profiles also showed that in environmental information processing, membrane transport (ABC transporters, ko02010) and signal transduction (two-component system, ko02020) are highly enriched in microbiomes. Membrane transport is a collection of mechanisms that regulate the passage of solutes, such as ions and small molecules, through a biofilm, which is a bilayer of lipids embedded in proteins. The regulation of crossing membranes is attributed to the permeability of selective membranes, a characteristic of biofilms that enables the separation of substances with diferent chemical properties. In other words, these membranes might be permeable to some substances but not to others42. Amongst these membranes, the ABC transporter pathway was highly enriched in soil microbiomes. ATP-binding box (ABC) transporters are universally existed in microorganisms such as bacteria and are one of the biggest protein families known today. These transporters bind ATP hydrolysis to participate in the active transport of multifarious substrates such as ions, peptides, lipids, drugs, sugars, proteins, and sterols. The structure of ABC transporters in prokaryotes usually comprises three parts. Generally, two intact membrane proteins each have six transmembrane fragments: two peripheral proteins that bind and hydrolyze ATP, and one peripheral (or lipoprotein) substrate of a binding protein. As observed in the genomes of many bacteria and archaea, many genes of these three components form operons43. Drought, salinity, and alkali stress promoted the membrane transport function, especially the improvement of the active transport function of soil microbiomes.
Signal transduction is a process in which a chemical or physical signal is transmitted through a cell as a series of molecular events. The most common is protein kinase-catalyzed protein phosphorylation. The two-component system is a signaling pathway that regulates many bacterial characteristics, such as toxicity, pathogenicity, symbiosis, motility, nutrient absorption, production of secondary metabolites, metabolic regulation, and cell division. These systems regulate physiological processes based on environmental or cellular parameters, enabling them to adapt to changing conditions44. The signal transduction of soil microbiomes was promoted by drought or saline-alkali stress.
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conclusion
This study is the first to present the soil microbiomes of the three ecotypes of C. deserticola. The following conclusions are obtained: (1) soil microbial community in grassland is the most abundant amongst the three habitats. (2) The biomarkers of the three ecotypes were also determined: Oceanospirillales (saline-alkali land), Sphingomonadales (grassland), and Propionibacteriales (sandy land). (3) Core microbiome analysis demonstrated that the soil microbial communities of C. deserticola were mostly had a drought, salt tolerance, alkali resistance, and stress resistance, such as Micrococcales and Bacillales. (4) The correlation analysis demonstrated that 2′-acetylacteoside is positively correlated with Oceanospirillales and echinacoside is significantly positively correlated with Sphingomonadales. (5) Tax4Fun predicts that the metabolic function profiles of three ecotypes of the soil microbiome are enriched in metabolism and environmental information processing.
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