Symbiotic Plants Research Articles

Isolation and characterization of endophytic bacteria from sterile leaf of Acrostichum aureum from Bengkalis Island (Riau, Indonesia) and its potency for antidiabetic

Abstract. Linda TM, Febriarti BL, Zul D, Sofyanti N, Berliansyah A, Delfira N, Devi S. 2023. Isolation and characterization of endophytic bacteria from sterile leaf of Acrostichum aureum from Bengkalis Island (Riau, Indonesia) and its potency for antidiabetic. Biodiversitas 24: 1580-1588. Endophytic bacteria are found in symbiotic plant tissues. These bacteria pose potential sources for the natural product due to their health benefit, e.g., for antidiabetic activity. The present study aimed to isolate and characterize endophytic bacteria isolates from the sterile leaf of sea fern (Acrostichum aureum Linn.) from Bengkalis Island (Riau Province), Indonesia. Furthermore, to investigate their potency as an antidiabetic agent. The characteristics of isolates bacteria were described based on colony morphology and biochemical tests. A total of six endophytic bacteria were isolated from the sterile leaf of Acrostichum aureum. Their characteristics vary in shape, color, and size. The phytochemical screening indicated the presence of alkaloids and saponin in all the isolates. The results exhibited that the extract of the sterile leaf of A. aureum also gave a similar phytochemical compound of isolate endophytic bacteria. The dendrogram constructed from morphological and biochemical characteristics showed the separation of D.SB 4.2 from other six bacteria strains due to the positive antibacterial activity for Escherichia coli ATCC35218, Staphylococcus aureus ATCC29213, Salmonella typhi ATCC14028, and Bacillus subtilis ATCC11774. Out of six isolates, only isolate D.SB 5.2 was selected for the antidiabetic test. The isolates showed positive results on the activity as a-amylase inhibitor (18.12%). This study provides important information on endophytic bacteria isolates on A. aureum and their potency for antidiabetic agents.

Diaporthe atlantica enhances tomato drought tolerance by improving photosynthesis, nutrient uptake and enzymatic antioxidant response.

Functional symbiosis with fungal endophytes can help plants adapt to environmental stress. Diaporthe atlantica is one of the most abundant fungal taxa associated with roots of Festuca rubra subsp. pruinosa, a grass growing in sea cliffs. This study aimed to investigate the ability of a strain of this fungus to ameliorate the impact of drought stress on tomato plants. In a greenhouse experiment, tomato plants were inoculated with Diaporthe atlantica strain EB4 and exposed to two alternative water regimes: well-watered and drought stress. Several physiological and biochemical plant parameters were evaluated. Inoculation with Diaporthe promoted plant growth in both water treatments. A significant interactive effect of Diaporthe-inoculation and water-regime showed that symbiotic plants had higher photosynthetic capacity, water-use efficiency, nutrient uptake (N, P, K, Fe and Zn), and proline content under drought stress, but not under well-watered conditions. In addition, Diaporthe improved the enzymatic antioxidant response of plants under drought, through an induced mechanism, in which catalase activity was modulated and conferred protection against reactive oxygen species generation during stress. The results support that Diaporthe atlantica plays a positive role in the modulation of tomato plant responses to drought stress by combining various processes such as improving photosynthetic capacity, nutrient uptake, enzymatic antioxidant response and osmo-protectant accumulation. Thus, drought stress in tomato can be enhanced with symbiotic fungi.

The wheat AMT2 (AMmonium Transporter) family, possible functions in ammonium uptake and pathogenic/symbiotic interactions

AbstractAmmonium uptake into wheat roots relies primarily on two AMmonium Transporters of subfamily one, while the wheat genome comprises 4 to 6 AMT2 type transporters. Plant AMT2s generally show functions in root‐to‐shoot translocation or pathogenic and symbiotic plant–microorganism interactions. We addressed the activity of TaAMT2s in ammonium transport. Nitrogen‐dependent expression implicated a physiological function in ammonium uptake for TaAMT2;1 and in ammonium distribution for TaAMT2;2‐6.

Adaptive bacterial and fungal matching between a parasitic plant and its host: A case of Cistanche deserticola and Haloxylon ammodendron

How the unique heterotrophic life history of parasitic plants influences the root and rhizosphere microbiota of symbiotic host plants is relatively unknown. In this study, we used Cistanche deserticola and Haloxylon ammodendron from cultivated populations as our model parasite and host plants, respectively. We collected samples from root (PR) and rhizosphere (PS) of the parasite, root (HIR) and rhizosphere (HIS) of the host plant infected, and root (HUR) and rhizosphere (HUS) of host plant uninfected. We conducted Illumina high-throughput DNA sequencing technology, and compared the diversity, composition, co-occurring network structure of their microbiota, and traced the composition of the parasitic plant microbiota. Our results showed that the diversity of root bacteria and fungi was lower than those in the rhizosphere, the co-association complexity of the root was simpler than those of the rhizosphere, but the compositional similarity of root microbiota was increased relative to those of the rhizosphere. Infection had localized and systemic effects on microbiota of HIR and HIS. Seven taxa were enriched in HIR and HIS, including Microvigra , Sphingomonas , Rubrobacter , Gaiella , Quadrisphaera , Chaetomium, and Penicillum . Additionally, infection also led to the co-association network of HIR being simpler than HUR. Meanwhile, the diversity, composition, and co-association network were similar between PR and HUR. Community assembly mechanisms analysis showed that the variable selection and homogenizing dispersal were highly similar between C. deserticola (Bacteria: 0.8 and 0.2; Fungi: 0.4 and 0.4) and H. ammodendron (Bacteria: 0.8 and 0.2; Fungi: 0.4 and 0.36). And PR displayed a high level of congruence with HUR in both bacterial ( t 0 =0.69) and fungal ( t 0 =0.53) communities. Collectively, our results suggested that the root microbiota of parasitic plant was highly congruent with those of the host plant, consistent with the phenomena of ‘parasitic equilibrium’. Our findings on parasite and host microbiota provide a new novel line of evidence supporting the complex interaction of parasitic plants and their hosts. • Lower diversity, and simpler co-association network but more similar composition were in root microbiota. • The infection had localized and systemic effects on the root and rhizosphere microbiota of H. ammodendron . • Variable selection and homogenizing dispersal were the main processes of bacterial and fungal community assembly. • High congruence in root microbiota between parasite and host reflected the phenomena of ‘parasitic equilibrium’.

Expression patterns of maize PIP aquaporins in middle or upper leaves correlate with their different physiological responses to drought and mycorrhiza.

Here we report the effect of Rhizophagus irregularis on maize leaf expression of six plasma membrane aquaporin isoforms from PIP1 and PIP2 subfamilies under severe drought development and recovery. The novelty of our study is the finding that leaf-specific mycorrhizal regulation of aquaporins is dependent on the position of the leaf on the shoot and changes in parallel with the rate of photosynthesis and the stomatal response to drought. The transcripts were isolated from the upper third (L3) or ear (L5) leaf, which differed greatly in physiological response to stress within each symbiotic variant. Aquaporins expression in upper L3 leaves appeared to be largely not sensitive to drought, regardless of symbiotic status. In contrast, L5 leaf of non-mycorrhizal plants, showed strong down-regulation of all PIPs. Mycorrhiza, however, protected L5 leaf from such limitation, which under maximal stress was manifested by 6-fold and circa 4-fold higher transcripts level for PIP1s and PIP2s, respectively. Distinct expression patterns of L3 and L5 leaves corresponded to differences in key parameters of leaf homeostasis - stomatal conductance, photosynthetic rates, and accumulation of ABA and SA as phytohormonal indicators of drought stress. In result symbiotic plants showed faster restoration of photosynthetic capability, regardless of leaf position, which we recognize as the hallmark of better stress tolerance. In summary, arbuscular mycorrhiza alleviates short-term drought effects on maize by preventing the down-regulation of plasma membrane aquaporins within middle leaves, thereby affecting stomatal conductance.

Chemical Ecology of Nematodes

Nematodes represent the most abundant group of metazoans on earth. They utilize diverse chemicals to interact with con-specific and hetero-specific organisms, and are also impacted by compounds produced by other interacting organisms. In the first part of this review we discuss how nematode-derived glycolipids modulate their behavior and development, as well as the interactions with other organisms. Furthermore, we provide a short overview about other secondary metabolites produced by nematodes that affect different life traits of free-living nematodes. In the second part of this review we discuss how different bacteria-, nematode-, and plant-derived chemicals such as volatile organic compounds, root exudates, and plant defenses regulate the interaction between entomopathogenic nematodes, their symbiotic bacteria, insect prey, predators, and plants.

Can Aphid Herbivory Induce Intergenerational Effects of Endophyte-conferred Resistance in Grasses?

Plants have evolved mechanisms to survive herbivory. One such mechanism is the induction of defences upon attack that can operate intergenerationally. Cool-season grasses (sub-family Pooideae) obtain defences via symbiosis with vertically transmitted fungal endophytes (genus Epichloë) and can also show inducible responses. However, it is unknown whether these herbivore-induced responses can have intergenerational effects. We hypothesized that herbivory by aphids on maternal plants induces the intergenerational accumulation of endophyte-derived defensive alkaloids and resistance intensification in the progeny. We subjected mother plants symbiotic or not with Epichloë occultans, a species known for its production of anti-insect alkaloids known as lolines, to the aphid Rhopalosiphum padi. Then, we evaluated the progeny of these plants in terms of loline alkaloid concentration, resistance level (through herbivore performance), and shoot biomass. Herbivory on mother plants did not increase the concentration of lolines in seeds but it tended to affect loline concentration in progeny plants. There was an overall herbivore-induced intergenerational effect increasing the endophyte-conferred defence and resistance. Symbiotic plants were more resistant to aphids and had higher shoot biomass than their non-symbiotic counterparts. Since maternal herbivory did not affect the loline concentrations in seeds, the greater resistance of the progeny could have resulted from an inherited mechanism of epigenetic regulation. It would be interesting to elucidate the origin of this regulation since it could come from the host or the fungal symbiont. Thus, endophyte-driven differential fitness between symbiotic and non-symbiotic plants might be higher as generations pass on in presence of herbivores.

Alder encroachment alters subsoil organic carbon pool and chemical structure in a boreal peatland of Northeast China

Boreal peatlands have been experiencing increased abundances of symbiotic dinitrogen-fixing woody plants (mainly alder species). However, how alder encroachment alters soil organic carbon (C) pool and stability is unclear. To examine the effects of alder encroachment on soil organic C, we measured soil organic C pool, phenol oxidase (POX) activity, organic C mineralization rate, and organic C chemical structure (alkyl C, O-alkyl C, aromatic C, and carbonyl C) using solid-state 13C nuclear magnetic resonance spectroscopy in the 0–10 cm, 10–20 cm, and 20–40 cm depths in the Alnus sibirica islands and adjacent open peatlands in the north of Da'xingan Mountain, Northeast China. A. sibirica islands had 28 %, 25 %, and 30 % greater POX activity and 36 %, 31 %, and 100 % higher organic C mineralization than open peatlands in the 0–10 cm, 10–20 cm, and 20–40 cm soil depths, respectively. Despite no significant changes in the 0–10 cm and 10–20 cm depths, alder encroachment reduced soil organic C pool in the 20–40 cm depth. Soil organic C pool in the 0–40 cm depth was lower in A. sibirica islands (298 Mg ha−1) than in the open peatlands (315 Mg ha−1). Moreover, alder encroachment increased alkyl (7 %) and carbonyl (57 %) C fractions but reduced O-alkyl C fraction (16 %) in the 20–40 cm depth, resulting in increased aliphaticity and recalcitrance indices. These findings suggest that alder encroachment will reduce soil organic C accumulation by accelerating microbial decomposition, and highlight that increased biochemical stabilization would attenuate soil organic C loss after alder expansion in boreal peatlands. Our results will help assess and project future C budgets in boreal peatlands.

Coinoculation impact on plant growth promotion: a review and meta-analysis on coinoculation of rhizobia and plant growth-promoting bacilli in grain legumes.

Coinoculation of symbiotic N2-fixing rhizobia and plant growth-promoting Bacillus on legume seeds can increase crop productivity. We collected highly resolved data on coinoculation of rhizobia and bacilli on 11 grain legume crops: chickpea, common bean, cowpea, faba bean, groundnut, lentil, mung bean, pea, pigeon pea, soybean, and urad bean to verify the magnitude of additive effects of coinoculation in relation to single inoculation of rhizobia on plant growth and yield of grain legumes. Coinoculation of rhizobia and bacilli on legume seeds and/or soil during sowing significantly increased nodulation, nitrogenase activity, plant N and P contents, and shoot and root biomass, as well as the grain yield of most grain legumes studied. There were however a few instances where coinoculation decreased plant growth parameters. Therefore, coinoculation of rhizobia and Bacillus has the potential to increase the growth and productivity of grain legumes, and can be recommended as an environmental-friendly agricultural practice for increased crop yields.

Epichloë endophyte affects the root colonization pattern of belowground symbionts in a wild grass

Plants host multiple symbionts that interact with each other affecting plant performance and regulating their establishment. Here, we analyzed how the association with Epichloë endophytes affects belowground colonization by Dark Septate Endophytes (DSE) and arbuscular mycorrhizal fungi (AMF) in the grass Bromus auleticus . Epichloë -symbiotic (E+) and Epichloë -non symbiotic (E−) plants were sampled from a long-term experimental plot and colonization structures were analyzed in the roots. We also examined the influence of Epichloë exudates on the in vitro growth of DSE Microdochium bolleyi isolated from roots. Epichloë symbiosis increased AMF colonization, although differences were not significant. Despite the lack of differences in total DSE colonization, in concordance with in vitro findings, a higher significant abundance of microsclerotia was observed in E+ plants. A negative correlation between total mycorrhizal and DSE was found. Our findings show a more uniform root colonization pattern in E+ plants, suggesting a root symbiosis modulating role. • DSE microsclerotia was significantly more abundant in B . auleticus E+ plants than E−. • Arbuscules abundance was double in plants associated with Epichloë compared to E−. • Epichloë endophytes exudates did not affect DSE strain growth in vitro . • DSE negatively correlate with AMF colonization. • E+ plants presents a more uniform root colonization pattern than Eplichloë -free.

Vertically Transmitted Epichloë Systemic Endophyte Enhances Drought Tolerance of Achnatherum inebrians Host Plants through Promoting Photosynthesis and Biomass Accumulation.

Achnatherum inebrians (drunken horse grass, DHG) plants, a dominant grass species in the arid and semi-arid regions of northwest China, symbiotic with an Epichloë fungal endophyte, is well adapted to drought. However, little is known about how the presence of the foliar Epichloë endophyte enhances the tolerance of DHG to drought at the molecular level. This study explored the positive effects of the presence of the Epichloë endophyte on plant growth, biomass, and photosynthetic efficiency and processes of DHG under non-drought and two drought (moderate and severe) treatments, using RNA sequencing to compare transcriptomes. The transcriptome results showed that 32 selected unigenes involved in the photosynthesis processes within Epichloë symbiotic plants were differently expressed (DEGs) versus non-symbiotic plants. The majority of these selected DEGs were upregulated in Epichloë symbiotic plants versus non-symbiotic plants, such as upregulated unigenes (c51525.graph_c1, c47798.graph_c0 & c64087.graph_c0) under drought conditions. In line with the transcriptomes data, the presence of the Epichloë endophyte promoted the photosynthetic rate and biomass accumulation of DHG plants, and the relationship between the photosynthetic rate and biomass is linear and significant. The presence of the endophyte only increased the biomass per tiller of DHG plants under drought. This study provides further insights into the molecular mechanisms that underlie the enhanced plant growth and drought tolerance of Epichloë-symbiotic DHG plants.

Endophytism: A Multidimensional Approach to Plant-Prokaryotic Microbe Interaction.

Plant growth and development are positively regulated by the endophytic microbiome via both direct and indirect perspectives. Endophytes use phytohormone production to promote plant health along with other added benefits such as nutrient acquisition, nitrogen fixation, and survival under abiotic and biotic stress conditions. The ability of endophytes to penetrate the plant tissues, reside and interact with the host in multiple ways makes them unique. The common assumption that these endophytes interact with plants in a similar manner as the rhizospheric bacteria is a deterring factor to go deeper into their study, and more focus was on symbiotic associations and plant–pathogen reactions. The current focus has shifted on the complexity of relationships between host plants and their endophytic counterparts. It would be gripping to inspect how endophytes influence host gene expression and can be utilized to climb the ladder of “Sustainable agriculture.” Advancements in various molecular techniques have provided an impetus to elucidate the complexity of endophytic microbiome. The present review is focused on canvassing different aspects concerned with the multidimensional interaction of endophytes with plants along with their application.

Deciphering the Potential Role of Symbiotic Plant Microbiome and Amino Acid Application on Growth Performance of Chickpea Under Field Conditions.

The unprecedented rise in the human population has increased pressure on agriculture production. To enhance the production of crops, farmers mainly rely on the use of chemical fertilizers and pesticides, which have, undoubtedly, increased the production rate but at the cost of losing sustainability of the environment in the form of genetic erosion of indigenous varieties of crops and loss of fertile land. Therefore, farming practices need to upgrade toward the use of biological agents to maintain the sustainability of agriculture and the environment. In this context, using microbial inoculants and amino acids may present a more effective, safer, economical, and sustainable alternative means of realizing higher productivity of crops. Therefore, field experiments were performed on chickpea for two succeeding years using Rhizobium and L-methionine (at three levels, i.e., 5, 10, and 15 mg L–1) separately and in combinations. The results show that the application of Rhizobium and all the three levels of L-methionine increased the growth and yield of chickpea. There was a higher response to a lower dose of L-methionine, i.e., 5 mg L–1. It has been found that maximum grain yield (39.96 and 34.5% in the first and second years, respectively) of chickpea was obtained with the combined use of Rhizobium and L-methionine (5 mg L–1). This treatment was also the most effective in enhancing nodule number (91.6 and 58.19%), leghemoglobin (161.1 and 131.3%), and protein content (45.2 and 45%) of plants in both years. Likewise, photosynthetic pigments and seed chemical composition were significantly improved by Rhizobium inoculation. However, these effects were prominent when Rhizobium inoculation was accompanied by L-methionine. In conclusion, utilizing the potential of combined use of L-methionine and microbial inoculant could be a better approach for developing sustainable agriculture production.

Germinating the Ant Plant

This article is an investigation into how short film ideas are conceived. It examines Ant Plant as a case study of inspiration based in bio-art, the Australian bushfires of 2020 and critical plant studies. The Ant Plant is set in an apocalyptic near future, in a burnt environment. The Guardian seeking to regenerate near-extinct species summons her brethren to deliver rare plants to her at the Greenhouse. A young woman hears the call and sets off on an arduous journey to deliver the key to a conserved plant kingdom – symbiotic Ant Plant.

Nickel mine soil is a potential source for soybean plant growth promoting and heavy metal tolerant rhizobia.

Mine soil is not only barren but also contaminated by some heavy metals. It is unclear whether some rhizobia survived under extreme conditions in the nickel mine soil. Therefore, this study tries to isolate some effective soybean plant growth promoting and heavy metal resistant rhizobia from nickel mine soil, and to analyze their diversity. Soybean plants were used to trap rhizobia from the nickel mine soil. A total of 21 isolates were preliminarily identified as rhizobia, which were clustered into eight groups at 87% similarity level using BOXA1R-PCR fingerprinting technique. Four out of the eight representative isolates formed nodules on soybean roots with effectively symbiotic nitrogen-fixing and plant growth promoting abilities in the soybean pot experiment. Phylogenetic analysis of 16S rRNA, four housekeeping genes (atpD-recA-glnII-rpoB) and nifH genes assigned the symbiotic isolates YN5, YN8 and YN10 into Ensifer xinjiangense and YN11 into Rhizobium radiobacter, respectively. They also showed different tolerance levels to the heavy metals including cadmium, chromium, copper, nickel, and zinc. It was concluded that there were some plant growth promoting and heavy metal resistant rhizobia with the potential to facilitate phytoremediation and alleviate the effects of heavy metals on soybean cultivation in nickel mine soil, indicating a novel evidence for further exploring more functional microbes from the nickel mine soil.

Emerging Roles of β-Glucanases in Plant Development and Adaptative Responses

Plant β-glucanases are enzymes involved in the synthesis, remodelling and turnover of cell wall components during multiple physiological processes. Based on the type of the glycoside bond they cleave, plant β-glucanases have been grouped into three categories: (i) β-1,4-glucanases degrade cellulose and other polysaccharides containing 1,4-glycosidic bonds to remodel and disassemble the wall during cell growth. (ii) β-1,3-glucanases are responsible for the mobilization of callose, governing the symplastic trafficking through plasmodesmata. (iii) β-1,3-1,4-glucanases degrade mixed linkage glucan, a transient wall polysaccharide found in cereals, which is broken down to obtain energy during rapid seedling growth. In addition to their roles in the turnover of self-glucan structures, plant β-glucanases are crucial in regulating the outcome in symbiotic and hostile plant–microbe interactions by degrading non-self glucan structures. Plants use these enzymes to hydrolyse β-glucans found in the walls of microbes, not only by contributing to a local antimicrobial defence barrier, but also by generating signalling glucans triggering the activation of global responses. As a counterpart, microbes developed strategies to hijack plant β-glucanases to their advantage to successfully colonize plant tissues. This review outlines our current understanding on plant β-glucanases, with a particular focus on the latest advances on their roles in adaptative responses.

White lupin (Lupinus albus L.) exposed to elevated atmospheric CO2 requires additional phosphorus for N2 fixation

PurposeThe steady rise of atmospheric CO2 concentrations enhances symbiotic N2 fixation and plant growth. However, it is largely unknown whether more P is required to enhance N2 fixation under elevated CO2. We examined if plants under elevated CO2 require more P to support enhanced N2 fixation.MethodsWhite lupin (Lupinus albus L. cv. Kiev) was grown in nutrient solution under ambient (380 ppm) and elevated (700 ppm) CO2 levels at 0, 2, 5, 12, 25 and 50 µM P. Nodulation, nodule function, N2 fixation and plant growth were assessed at 4 and 6 weeks post inoculation. Excess cation uptake and proton release were also analysed due to its importance in P acquisition.ResultsElevated CO2 increased the critical P concentration in nutrient solution for N2 fixation, indicating plants under elevated CO2 need to acquire additional P to support greater N2 fixation. Elevated CO2 improved N2 fixation in P-deficient plants by increasing specific nitrogenase activity. Elevated CO2 also enhanced excess cation uptake which increased proton release by roots.ConclusionsAs elevated CO2 increased the plant demand for P, it increased the critical P concentration in nutrient solution for N2 fixation, thereby plants require more P to mitigate the effect of P deficiency on N2 fixation. Despite this, elevated CO2 could enhance N2 fixation in P-limiting environments.

Expanding the Biological Role of Lipo-Chitooligosaccharides and Chitooligosaccharides in Laccaria bicolor Growth and Development.

The role of lipo-chitooligosaccharides (LCOs) as signaling molecules that mediate the establishment of symbiotic relationships between fungi and plants is being redefined. New evidence suggests that the production of these molecular signals may be more of a common trait in fungi than what was previously thought. LCOs affect different aspects of growth and development in fungi. For the ectomycorrhizal forming fungi, Laccaria bicolor, the production and effects of LCOs have always been studied with a symbiotic plant partner; however, there is still no scientific evidence describing the effects that these molecules have on this organism. Here, we explored the physiological, molecular, and metabolomic changes in L. bicolor when grown in the presence of exogenous sulfated and non-sulfated LCOs, as well as the chitooligomers, chitotetraose (CO4), and chitooctaose (CO8). Physiological data from 21 days post-induction showed reduced fungal growth in response to CO and LCO treatments compared to solvent controls. The underlying molecular changes were interrogated by proteomics, which revealed substantial alterations to biological processes related to growth and development. Moreover, metabolite data showed that LCOs and COs caused a downregulation of organic acids, sugars, and fatty acids. At the same time, exposure to LCOs resulted in the overproduction of lactic acid in L. bicolor. Altogether, these results suggest that these signals might be fungistatic compounds and contribute to current research efforts investigating the emerging impacts of these molecules on fungal growth and development.

Phylogenetic diversity and plant growth-promoting activities of rhizobia nodulating fenugreek (Trigonella foenum-graecum Linn.) cultivated in different agroclimatic regions of India.

Fenugreek (Trigonella foenum-graecum Linn.), is an extensively cultivated legume crop used as a herb, spice, and traditional medicine in India. The symbiotic efficiency and plant growth-promoting potential of fenugreek rhizobia depend on the symbiont strain and environmental factors. We isolated 176 root-nodulating bacteria from fenugreek cultivated in different agroclimatic regions of India. MALDI-TOF MS-based identification and phylogenetic analyses based on 16S rRNA and five housekeeping genes classified the fenugreek-rhizobia as Ensifer (Sinorhizobium) meliloti. However, the strains represent separate sub-lineages of E. meliloti, distinct from all reported sub-lineages across the globe. We also observed the spatial distribution of fenugreek rhizobia, as the three sub-lineages of E. meliloti recorded during this study were specific to their respective agroclimatic regions. According to the symbiotic gene (nodC and nifH) phylogenies, all three sub-lineages of E. meliloti harboured symbiotic genes similar to symbiovar meliloti; as with the housekeeping genes, these also revealed a spatial distribution for different clades of sv. meliloti. The strains could nodulate fenugreek plants and they showed plant growth-promoting potential. Significant differences were found in the plant growth parameters in response to inoculation with the various strains, suggesting strain-level differences. This study demonstrates that fenugreek rhizobia in India are diverse and spatially distributed in different agro-climatic regions.

Quantitative transcriptomic and metabolomic analyses reveal the changes in Tricholoma matsutake fruiting bodies during cold storage

A combination of transcriptomic and metabolomic analyses was performed to systematically understand the metabolic changes in Tricholoma matsutake fruiting bodies during cold storage. In total, 800 metabolites were identified and 19,964 annotated unigenes were quantified. The unigenes related to the catabolism of proteins, carbohydrates, and lipids were mainly upregulated during cold storage, but the related primary metabolites were not accumulated, which indicated complete degradation and loss of nutrients. Concurrently, the synthesis and metabolism of the main components of the cell wall, chitin and β-1,3-glucan, were regulated, indicating the dynamic remodeling of the T. matsutake cell wall structure. Additionally, indole-3-acetic acid and components of its synthesis pathway were found in T. matsutake, indicating their potential role as a communicator between T. matsutake and its symbiotic plants. The results provide new information to improve the understanding of the metabolic mechanism of T. matsutake fruit bodies during postharvest cold storage.