Biofilms are indispensable ecological habitats for microbes that have garnered global attention and play a potential role in influencing the biogeochemical cycling of nitrogen. However, the biogeochemical significance of biofilms and the mechanisms by which they regulate nitrogen cycling remain elusive. In this study, we utilized DNA-stable isotope probing (DNA-SIP) labelling techniques in conjunction with metagenomics to reveal a nitrifying ecological niche in biofilms taken from the Yangtze Estuary, with those from sediment and water samples for comparison. Quantitative analysis showed that the amoA gene abundance of comammox Nitrospira (2.3 × 103 copies ng-1 DNA) was significantly higher than that of ammonia-oxidizing archaea (AOA-amoA, 62.4 copies ng-1 DNA) and ammonia-oxidizing bacteria (AOB-amoA, 218.1 copies ng-1 DNA) in biofilms, and the average abundance of comammox Nitrospira showed the following order: water > biofilm > sediment. Moreover, the NOB nxrB gene was more abundant than the amoA gene of ammonia oxidizers in all three media. DNA-SIP further revealed that the active comammox Nitrospira clade A mediates the nitrification process in biofilms with peak abundance at a buoyant density of 1.715 g mL-1. Active nitrifying bacteria exhibit metabolic diversity in both biofilms and sediments, and occupy unique nitrifying ecological niches. Additionally, the co-occurrence network showed that chlorophyll a, NO3- and salinity emerged as the predominant physicochemical factors affecting the nitrogen transformation genes in biofilms. Taken together, this study indicates that biofilms constitute an emerging nitrifying ecological niche in estuarine environments and deepens our understanding of the mechanisms by which biofilms function in marine biogeochemistry.

Dissolved organic matter (DOM) is involved in numerous biogeochemical processes, and understanding the ecological succession of DOM is crucial for predicting its response to farming (e.g., fertilization) practices. Although plentiful studies have examined how fertilization practice affects the content of soil DOM, it remains unknown how long-term fertilization drives the succession of soil DOM over temporal scales. Here, we investigated the succession of DOM in paddy rice rhizosphere soils subjected to different long-term fertilization treatments (CK: no fertilization; NPK: inorganic fertilization; OM: organic fertilization) along with plant growth. Our results demonstrated that long-term fertilization significantly promoted the molecular chemodiversity of DOM, but it weakened the correlation between DOM composition and plant development. Time-decay analysis indicated that the DOM composition had a shorter halving time under CK treatment (94.7 days), compared to NPK (337.4 days) and OM (223.8 days) treatments, reflecting a lower molecular turnover rate of DOM under fertilization. Moreover, plant development significantly affected the assembly process of DOM only under CK, not under NPK and OM treatments. Taken together, our results demonstrated that long-term fertilization, especially inorganic fertilization, greatly weakens the ecological succession of DOM in the plant rhizosphere, which has a profound implication for understanding the complex plant-DOM interactions.

Plant-associated microorganisms are believed to be part of the so-called extended plant phenotypes, affecting plant growth and health. Understanding how plant-associated microorganisms respond to pathogen invasion is crucial to controlling plant diseases through microbiome manipulation. In this study, healthy and diseased (bacterial wilt disease, BWD) tomato (Solanum lycopersicum L.) plants were harvested, and variations in the rhizosphere and root endosphere microbial communities were subsequently investigated using amplicon and shotgun metagenome sequencing. BWD led to a significant increase in rhizosphere bacterial diversity in the rhizosphere but reduced bacterial diversity in the root endosphere. The ecological null model indicated that BWD enhanced the bacterial deterministic processes in both the rhizosphere and root endosphere. Network analysis showed that microbial co-occurrence complexity was increased in BWD-infected plants. Moreover, higher universal ecological dynamics of microbial communities were observed in the diseased rhizosphere. Metagenomic analysis revealed the enrichment of more functional gene pathways in the infected rhizosphere. More importantly, when tomato plants were infected with BWD, some plant-harmful pathways such as quorum sensing were significantly enriched, while some plant-beneficial pathways such as streptomycin biosynthesis were depleted. These findings broaden the understanding of plant-microbiome interactions and provide new clues to the underlying mechanism behind the interaction between the plant microbiome and BWD.

Exploiting the potential benefits of plant-associated microbes represents a sustainable approach to enhancing crop productivity. Plant-beneficial bacteria (PBB) provide multiple benefits to plants. However, the biogeography and community structure remain largely unknown. Here we constructed a PBB database to couple microbial taxonomy with their plant-beneficial traits and analysed the global atlas of potential PBB from 4,245 soil samples. We show that the diversity of PBB peaks in low-latitude regions, following a strong latitudinal diversity gradient. The distribution of potential PBB was primarily governed by environmental filtering, which was mainly determined by local climate. Our projections showed that fossil-fuel-dependent future scenarios would lead to a significant decline of potential PBB by 2100, especially biocontrol agents (-1.03%) and stress resistance bacteria (-0.61%), which may potentially threaten global food production and (agro)ecosystem services.

Dissolved organic matter (DOM) is involved in numerous biogeochemical processes, and its molecular weight affects many of these processes through its bioavailability and sorptive capacity. However, it remains unknown to what extent the molecular weight of DOM mediates its dynamics, for example, influencing its role in DOM-microbe interactions and the processes determining the compositional assembly of DOM. To address this issue, ultrahigh-resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) and high-throughput sequencing were applied to investigate how the molecular weight of DOM was associated with its dynamics in two typical agricultural soils with different fertility. Our results showed that low-molecular-weight DOM had lower biological stability and a higher transformation potential. Analysis of the DOM-microbe co-occurrence network showed that low-molecular-weight DOM displayed tighter interactions with a diversity of microbes, while high-molecular-weight DOM interacted with only a few microbes. Ecological null models revealed that the compositional assembly of low-molecular-weight DOM, but not high-molecular-weight DOM, was more controlled by deterministic processes. Taken together, our results demonstrate the fundamental role the molecular weight of DOM plays in determining biological stability, transformation potential, interactions with microbes, and assembly mechanisms of DOM in agricultural soils. This work provides the foundation for general principles explaining complex dynamics of DOM in natural ecosystems, highlighting that using theories and concepts in metacommunity ecology, such as community diversity and assembly mechanisms, may open a new avenue to understand DOM dynamics from a macro perspective.

Phytopathogenic fungi threaten global food security but the ecological drivers of their global diversity and biogeography remain unknown. Here, we construct and analyse a global atlas of potential phytopathogenic fungi from 20,312 samples across all continents and major oceanic island regions, eleven land cover types, and twelve habitat types. We show a peak in the diversity of phytopathogenic fungi in mid-latitude regions, in contrast to the latitudinal diversity gradients observed in aboveground organisms. Our study identifies climate as an important driver of the global distribution of phytopathogenic fungi, and our models suggest that their diversity and invasion potential will increase globally by 2100. Importantly, phytopathogen diversity will increase largely in forest (37.27-79.12%) and cropland (34.93-82.51%) ecosystems, and this becomes more pronounced under fossil-fuelled industry dependent future scenarios. Thus, we recommend improved biomonitoring in forests and croplands, and optimised sustainable development approaches to reduce potential threats from phytopathogenic fungi.

以青藏高原3种典型生境(荒漠草地、湿地和盐碱地)为研究对象,通过amoA功能基因Qpcr和 16S Rrna基因扩增子测序,研究了其氨氧化古菌(ammonia-oxidizing archaea,AOA)、氨氧化细菌(ammonia-oxidizing bacteria,AOB)、亚硝酸盐氧化菌(nitrite-oxidizing bacteria,NOB)和完全硝化细菌(complete ammonia oxidizer,CMX)的分布与群落结构特征.Qpcr结果表明,荒漠草地和盐碱地中 3 种氨氧化微生物,其丰度顺序为AOA>CMX>AOB,而在湿地中则为CMX≥AOA>AOB.各生境中AOA主要类群为土壤类群(即group 1.1b),且其中约半数属于Nitrosocosmicus属分支,该分支在北极冻土中也有分布.荒漠草地和湿地中AOB主要为亚硝化螺菌属(Nitrosospira spp.,71.21％～100％),而盐碱地主要为亚硝化单胞菌属(Nitrosomonas spp.,75.51％～88.71％).3 种生境中NOB主要类群均为硝化螺菌属(Nitrospira spp.,70.87％～98.79％).clade A和clade B分支的CMX在各生境均存在,且二者比例相当.值得注意的是,大多生境中都检测出一种或多种典型海洋类群硝化微生物,比如group 1.1a的AOA、N.marina的AOB、Lineage 4 的NOB,这强烈暗示青藏高原硝化微生物可能受青藏高原古海洋环境的影响.

氨氧化古菌(Ammonia-oxidizing archaea,AOA)被认为是酸性土壤硝化过程的主要微生物类群,但AOA如何适应酸性胁迫并发挥作用一直是研究难点,而ATP酶(ATPase)是能量代谢的关键,其编码基因可能在AOA适应酸性胁迫过程中发生了趋同性演化.据此,本研究针对5个不同种植年限的马尾松人工林酸性土壤(15 a、24 a、45 a、55 a、63 a),通过深度宏基因组测序获得7360亿碱基对,重构AOA氨单加氧酶amoA基因和ATP酶A亚基(ATPase subunit A)基因的系统发育进化谱系,研究AOA适酸的分子机制.结果表明:根据经典的amoA基因系统发育进化分类,所有5个森林土壤中优势AOA主要包括Nitrososphaerales和Ca.Nitrosotaleales两大类群,但Nitrososphaerales类群与中碱性土壤中的AOA古菌亲缘关系更近,与嗜酸的Ca.Nitrosotaleales类群亲缘关系较远,表明amoA基因的系统进化关系不能解释Nitrososphaerales在酸性土壤中的成功定殖.然而,基于ATPase subunit A基因的系统进化分析则发现,所有酸性森林土壤中嗜酸/耐酸氨氧化古菌均含有亲缘关系较近的V-ATPase subunit A基因,表明氨氧化古菌可能通过基因水平转移获得V-ATPase基因适应酸性胁迫环境,较好地解释了氨氧化古菌适应酸性胁迫的生境扩展规律.随林龄的增加,Ca.Nitrosotaleales类群丰度先减少后增加,而Nitrososphaerales类群丰度先增加后减少,速效钾是显著影响AOA群落结构的重要环境因子.这些结果表明,不同种植年限下酸性人工林土壤中氨氧化古菌种群发生了明显的分化,V-ATPase基因水平转移可能是氨氧化古菌适应酸性胁迫的重要机制.

Body size determines individuals' life history and metabolic rates and thus, regulates community-level dynamics. However, whether body size mediates community co-occurrences and stability, especially in complex communities across different microbial trophic levels, remains unknown. Here, we investigate whether body size determines the co-occurrence pattern and stability of microbial communities across local, regional, and continental scales in the paddy soil ecosystems. Soil samples were collected from rice paddy fields at multiple spatial scales, and soil microbial communities were subsequently sequenced. The microorganisms were then divided into different groups based on taxonomic information at phylum/subphylum level, and the average body size of each microbial group was identified based on propagule size fromdocumented literature. We examined the relationships betweenmicrobial body size and various community traits such as potential migration rate, co-occurrence pattern, cohesion, and community stability. Our results consistently showed that the small-sized microorganisms such as bacteria had significantly higher niche breadth, niche overlap and migration rate at various spatial scales. We found that microbial body size is consistently negatively correlated to negative co-occurrences and community stability. Our results, for the first time, put microbial body size into a broader community ecology framework, and contribute to a greater understanding of how microbial taxa with different body sizes would respond to future changes and perturbations.

The discovery of complete ammonia oxidation (comammox), oxidizing ammonia to nitrate via nitrite in a single organism, has redefined the traditional recognition of the two-step nitrification driven by two functional groups (ammonia-oxidizing and nitrite-oxidizing microorganisms). However, the understanding of the distribution and niche differentiation of comammox Nitrospira in the estuarine mudflats and their reclaimed agricultural soils is still limited. Here, we investigated the abundance, diversity and community structures of comammox Nitrospira in the mudflats and the reclaimed agricultural soils in the northern Yangtze River estuary. Quantitative PCR showed the abundances of amoA genes of comammox were lower than that of ammonia-oxidizing bacteria (AOB) in nearly all samples. Amplicon sequencing of amoA genes revealed that the community structures of comammox Nitrospira were significantly ( P < 0.001) different between the original mudflats and the reclaimed agricultural soils, indicating niche differentiation among comammox Nitrospira clades (clade A.1, clade A.2, and clade B). The clade A.1 was the dominant group of comammox Nitrospira in the mudflats, while clade B predominated in the agricultural soils. However, the members of clade A.2 could be clearly divided into two groups, the mudflat-preferred and agricultural soil-preferred groups, suggesting more complicated ecological preferences within this sub-clade. Furthermore, it was demonstrated that salinity, organic matter (OM) and NO 3 – -N had a significantly influence on the distribution of comammox Nitrospira in the estuarine environment. Clade A.1 and nearly half members of clade A.2 were positively correlated with salinity, and negatively correlated with the concentrations of OM and NO 3 – -N. In contrast, the clade B and the other half members of clade A.2 showed the exact opposite pattern: a negative correlation with salinity and positive correlation with OM and NO 3 – -N. The co-occurrence network demonstrated that the operational taxonomic units (OTUs) within the same (sub-)clade were mostly positively correlated, indicating the similar niche preferences among the members from the same (sub-)clade of comammox Nitrospira . Taken together, our results revealed the niche differentiation of comammox Nitrospira in estuarine ecosystems where salinity and OM were the primary factors responsible for the distinct ecological distribution patterns.

全球30％以上陆地面积是酸性土壤(pH<5.5),而酸性土壤中氨氧化微生物群落特征研究是破译其硝化过程微生物学机理的基础.尤其随着完全硝化微生物(Complete ammonia oxidizer,comammox)的发现,亟需重新认知酸性土壤中氨氧化微生物类群.以酸性马尾松林为研究对象,综合利用荧光定量PCR(qPCR)、凝胶电泳半定量和宏基因组测序等技术研究土壤中氨氧化古菌(Ammonia-oxidizing archaea,AOA)、氨氧化细菌(Ammonia-oxidizing bacteria,AOB)和Comammox的相对丰度以及群落组成特征.研究发现AOA和AOB amoA基因丰度分别为2.61×106 copies·g–1和1.45×106 copies·g–1;而comammox amoA基因qPCR结果存在显著的非特异性扩增,导致其丰度被高估,而经凝胶电泳半定量矫正后,约为(1.38～1.47)×106 copies·g–1,该结果和土壤宏基因测序揭示的comammox相对丰度基本吻合.此外,宏基因组分析发现经典嗜酸group 1.1a-associated仅占AOA总类群的12％,而group 1.1b则占88％,尽管目前仍未有嗜酸group 1.1b AOA纯菌株的报道.AOB主要类群为Nitrosospira(约64％),而Nitrosomonas约占36％.Comammox主要类群为clade B(约64％),而clade A仅占36％且均隶属于clade A.1亚枝,这暗示clade B与已报道的嗜中性comammox clade A纯菌株有极大的生理代谢差异.总之,本研究提供了综合利用qPCR、半定量和宏基因组分析土壤氨氧化微生物群落的策略,并建议优化comammox的qPCR引物,同时本研究系统分析了酸性马尾松林土壤中氨氧化微生物的相对丰度和群落组成特征.

After the discovery of complete ammonia-oxidizing (comammox) Nitrospira, detection and assessments of the contribution of comammox Nitrospira communities to nitrogen cycling are in great demand. PCR-based approach, a common method for the detection of comammox, depends strongly on accurate amplification of the amoA genes from the original DNA samples using appropriate primers. In this study, we reported an evaluation of the performance of two commonly used primer sets, Ntsp-amoA 162F/359R and comaA/B-244f/659r, for amplifying the comammox amoA genes from three representative wetland soils in China [Sangsang (SS), Sanjiang (SJ), and Xianghai (XH)]. Our results demonstrated the two primer sets could both successfully amplify the clades with high relative abundances (RA), and further revealed a broadly similar diversity and community composition of dominant comammox operational taxonomic units (OTUs) (RA ≥ 1%) in each of the three wetland soils. However, the clades with low RA, such as the clade A (1.26%) in SJ and the clade B (11.54%) in XH that were recovered by metagenomics analysis, failed to be amplified using comaA/B-244f/659r, but were successfully amplified and sequenced using Ntsp-amoA 162F/359R. It indicated that, compared to comaA/B-244f/659r, Ntsp-amoA 162F/359R was more sensitive to the clades with low RA. However, it is worth noting that Ntsp-amoA 162F/359R would overestimate the RA of some rare clades. For example, the RAs of clade B in XH were overestimated by 32-fold. Furthermore, high levels of non-target amplification were detected via gel electrophoresis using both primer sets, especially for comammox Clade B amoA genes, implying that we should treat qPCR results based on these primers with caution. Taken together, our study comprehensively compared the performance of the two primer sets on the sensitivity and specificity of amplifying comammox amoA genes in three wetland soils, pointing out the necessity of further development of new primers for the efficient and accurate detection of comammox in various environments.

Zinc (Zn) is an essential element for plant growth and development, and Zn derived from crop plants in the diet is also important for human health. Here, we report that genetic variation in Heavy Metal-ATPase 4 (HMA4) controls natural variation in leaf Zn content. Investigation of the natural variation in leaf Zn content in a world-wide collection of 349 Arabidopsis thaliana wild collected accessions identified two accessions, Van-0 and Fab-2, which accumulate significantly lower Zn when compared with Col-0. Both quantitative trait loci (QTL) analysis and bulked segregant analysis (BSA) identified HMA4 as a strong candidate accounting for this variation in leaf Zn concentration. Genetic complementation experiments confirmed this hypothesis. Sequence analysis revealed that a 1-bp deletion in the third exon of HMA4 from Fab-2 is responsible for the lose of function of HMA4 driving the low Zn observed in Fab-2. Unlike in Fab-2 polymorphisms in the promoter region were found to be responsible for the weak function of HMA4 in Van-0. This is supported by both an expression analysis of HMA4 in Van-0 and through a series of T-DNA insertion mutants which generate truncated HMA4 promoters in the Col-0 background. In addition, we also observed that Fab-2, Van-0 and the hma4-2 null mutant in the Col-0 background show enhanced resistance to a combination of high Zn and high Cd in the growth medium, raising the possibility that variation at HMA4 may play a role in environmental adaptation.