Nitrogen Fixation Capacity Research Articles

The Mesorhizobium huakuii transcriptional regulator AbiEi plays a critical role in nodulation and is important for bacterial stress response

BackgroundBacterial abortive infection (Abi) systems are type IV toxin–antitoxin (TA) system, which could elicit programmed cell death and constitute a native survival strategy of pathogenic bacteria under various stress conditions. However, no rhizobial AbiE family TA system has been reported so far. Here, a M. huakuii AbiE TA system was identified and characterized.ResultsA mutation in M. huakuii abiEi gene, encoding an adjacent GntR-type transcriptional regulator, was generated by homologous recombination. The abiEi mutant strain grew less well in rich TY medium, and displayed increased antioxidative capacity and enhanced gentamicin resistance, indicating the abiEi operon was negatively regulated by the antitoxin AbiEi in response to the oxidative stress and a particular antibiotic. The mRNA expression of abiEi gene was significantly up-regulated during Astragalus sinicus nodule development. The abiEi mutant was severely impaired in its competitive ability in rhizosphere colonization, and was defective in nodulation with 97% reduction in nitrogen-fixing capacity. The mutant infected nodule cells contained vacuolation and a small number of abnormal bacteroids with senescence character. RNA-seq experiment revealed it had 5 up-regulated and 111 down-regulated genes relative to wild type. Of these down-regulated genes, 21 are related to symbiosis nitrogen fixation and nitrogen mechanism, 16 are involved in the electron transport chain and antioxidant responses, and 12 belong to type VI secretion system (T6SS).ConclusionsM. huakuii AbiEi behaves as a key transcriptional regulator mediating root nodule symbiosis.

Testing Whether Pre-Pod-Fill Symbiotic Nitrogen Fixation in Soybean Is Subject to Drift or Selection Over 100 Years of Soybean Breeding

Soybean [Glycine max (L.) Merr.] is the world's leading legume crop and the largest oilseed crop. It forms a symbiotic relationship with rhizobia bacteria residing in root nodules that provide fixed nitrogen to host plants through symbiotic nitrogen fixation (SNF). In soybean, it has been widely reported that the highest SNF occurs at the pod-filling stage, associated with the peak demand for nitrogen. However, the majority of seed nitrogen is derived from remobilizing root/shoot nitrogen, representing cumulative SNF from the seedling stage to the pre-pod-fill stage. Therefore, the question arises as to whether there has also been selection for improved SNF at these earlier stages, or whether pre-pod-fill SNF traits have drifted. To test this hypothesis, in this study, pre-pod SNF-related traits were evaluated in soybean cultivars that span 100 years of breeding selection in the Canadian Province of Ontario. Specifically, we evaluated SNF traits in 19 pedigree-related historical cultivars and 25 modern cultivars derived from the University of Guelph soybean breeding program. Field trials were conducted at Woodstock, Ontario, Canada in 2016 and 2017, and various SNF-related traits were measured at pre-pod-fill stages (R1-R3), including nitrogen fixation capacity. Considerable variation was observed among Canadian soybean cultivars released over the past 100 years for pre-pod-fill nitrogen fixation. The modern soybean cultivars had similar or moderately higher pre-pod-fill SNF compared to the historical lines in terms of the percentage of nitrogen derived from the atmosphere (%Ndfa) and total shoot fixed nitrogen. These findings suggest that, despite no direct selection by breeders, pre-pod-fill nitrogen fixation, and associated SNF traits have been maintained and possibly improved in modern soybean breeding. However, the low level of pre-pod-fill SNF in some modern cultivars, and generally wide variation observed in SNF between them, suggest some level of genetic drift for this trait in some pedigrees. Specific historical and modern soybean cultivars were identified as potential parents to enable targeted breeding for improved pre-pod-fill SNF. This retrospective study sheds light on our understanding of the impact of decades of recent selective breeding on pre-pod-fill nitrogen fixation traits in soybean in a temperate environment.

Proteomic mechanisms for the regulation of growth, photosynthetic activity and nitrogen fixation in Nostoc sp. PCC 7120 exposed to three antibiotic contaminants

This study investigated the influences of three frequently detected antibiotics in surface waters, ciprofloxacin, tetracycline and sulfamethoxazole, on the growth, photosynthetic activity, nitrogen-fixing capacity and proteomic expression profiles of Nostoc sp. PCC 7120, through a 15-day exposure test at environmentally relevant exposure doses of 50–200 ng/L. Cyanobacterial growth was stimulated by 100 ng/L and 200 ng/L of ciprofloxacin and sulfamethoxazole as well as 50–200 ng/L of tetracycline. The nitrogenase synthesis ability in each cyanobacterial cell was stimulated by 50–200 ng/L of ciprofloxacin while inhibited by 100 ng/L and 200 ng/L of tetracycline and sulfamethoxazole. At the exposure dose of 100 ng/L for each antibiotic, the variation of total nitrogen in the culture medium indicated that the nitrogen-fixing capacity of Nostoc sp. was determined by total nitrogenase concentration calculated by cell density × nitrogenase synthesis ability. Therefore, ciprofloxacin enhanced nitrogen fixation through the stimulation of both cyanobacterial growth and nitrogenase synthesis, while tetracycline and sulfamethoxazole enhanced nitrogen fixation merely through growth stimulation. At the exposure dose of 100 ng/L, only two downregulated proteins, a phosphonate ABC transporter and a methionine aminopeptidase, as well as one upregulated protein, the phenylalanine-tRNA ligase alpha subunit, were commonly shared by three antibiotic-treated groups. Ciprofloxacin upregulated proteins related to nitrogen fixation, carbon catabolism and biosynthesis, but downregulated photosynthesis-related proteins. In contrast, tetracycline and sulfamethoxazole increased the photosynthetic activity of Nostoc sp. through upregulating photosynthesis-related proteins, but downregulated proteins related to nitrogen fixation, carbon catabolism and biosynthesis. The resistance of Nostoc sp. PCC 7120 to three target antibiotics were related with the responses of RNA synthesis regulatory proteins. Stimulation of cyanobacterial nitrogen fixation by antibiotic contaminants could aggravate eutrophication in aquatic environments.

Assessment of the production and importance of cowpea [Vigna unguiculata (L.) walp]: Cases from selected districts of southern Ethiopia

Cowpea (Vigna unguiculata L.Walp) is an important legume in the hot, dry tropics and subtropics of sub-Saharan Africa, serving a multiple role for the livelihoods of millions of relatively low-income people. The entire plant can be used for either human or livestock consumption and with considerable drought-tolerating capacity. Tender young leaves, green pods and matured seeds are used as human food. Moreover, the crop serves for sustainable soil fertility improvement due to its excellent nitrogen-fixing capacity. However, its production and utilization are limited in Ethiopia partly due to dependence on the conventional agronomic practices and lack of information on its wide ranging uses. This study was conducted to assess the cowpea agronomy and the contributions the crop has in the livelihoods of farmers at Loka-Abaya and Humbo districts of Southern Ethiopia. Multi-stage sampling techniques were employed to achieve the set objectives. Both primary and secondary data were collected to solicit the required information. The data were subjected to descriptive and inferential statistics such as multiple linear regression model using the SPSS Software version 20 and STATA 13. Multiple linear regression model results showed that education, land size, climate information access, credit access, lack of market chain, availability of seed of improved varieties, and pests significantly (P<0.001) affected cowpea production in the studied areas. The trend analysis showed that the cowpea yield and production area coverage is increasing in Humbo District whereas, a decreasing trend was observed at the Loka Abaya. According to the household interview data, about 76 % of the respondents reported a decrease in the cultivated area of cowpea. According to the respondents, lack of access to improved seed and lack of extension support services contributed 79 % and 73 %, respectively to the low yield observed in the area. The majority of the respondents cultivate cowpea as intercropping and rotation with cereals and in the main field with the main purpose to replenish soil fertility (97 %). On the other hand, 62 % of the respondents cultivate cowpea for home consumption. According to the survey result, 48 % of the respondents use the matured grain for consumption. The production trends of the cowpea are highly variable mainly due to less attention paid by the extension systems to boost the yield of the crop, reliance of farmers on local varieties, pest occurrence and poor market chain. Therefore, modern production technologies including the supply of improved varieties of seed with their full production package should be introduced to the area so as to improve the yield and optimize its contribution towards achieving food security.

Genomic and molecular evidence reveals novel pathways associated with cell surface polysaccharides in bacteria.

Amino acid (acyl carrier protein) ligases (AALs) are a relatively new family of bacterial amino acid adenylating enzymes with unknown function(s). Here, genomic enzymology tools that employ sequence similarity networks and genome context analyses were used to hypothesize the metabolic function(s) of AALs. In over 50% of species, aal and its cognate acyl carrier protein (acp) genes, along with three more genes, formed a highly conserved AAL cassette. AAL cassettes were strongly associated with surface polysaccharide gene clusters in Proteobacteria and Actinobacteria, yet were prevalent among soil and rhizosphere-associated α- and β-Proteobacteria, including symbiotic α- and β-rhizobia and some Mycolata. Based on these associations, AAL cassettes were proposed to encode a noncanonical Acp-dependent polysaccharide modification route. Genomic-inferred predictions were substantiated by published experimental evidence, revealing a role for AAL cassettes in biosynthesis of biofilm-forming exopolysaccharide in pathogenic Burkholderia and expression of aal and acp genes in nitrogen-fixing Rhizobium bacteroids. Aal and acp genes were associated with dltBD-like homologs that modify cell wall teichoic acids with d-alanine, including in Paenibacillus and certain other bacteria. Characterization of pathways that involve AAL and Acp may lead to developing new plant and human disease-controlling agents as well as strains with improved nitrogen fixation capacity.

Differential responses of the sunn4 and rdn1-1 super-nodulation mutants of Medicago truncatula to elevated atmospheric CO2.

Nitrogen fixation in legumes requires tight control of carbon and nitrogen balance. Thus, legumes control nodule numbers via an autoregulation mechanism. 'Autoregulation of nodulation' mutants super-nodulate are thought to be carbon-limited due to the high carbon-sink strength of excessive nodules. This study aimed to examine the effect of increasing carbon supply on the performance of super-nodulation mutants. We compared the responses of Medicago truncatula super-nodulation mutants (sunn-4 and rdn1-1) and wild type to five CO2 levels (300-850 μmol mol-1). Nodule formation and nitrogen fixation were assessed in soil-grown plants at 18 and 42 d after sowing. Shoot and root biomass, nodule number and biomass, nitrogenase activity and fixed nitrogen per plant of all genotypes increased with increasing CO2 concentration and reached a maximum at 700 μmol mol-1. While the sunn-4 mutant showed strong growth retardation compared with wild-type plants, elevated CO2 increased shoot biomass and total nitrogen content of the rdn1-1 mutant up to 2-fold. This was accompanied by a 4-fold increase in nitrogen fixation capacity in the rdn1-1 mutant. These results suggest that the super-nodulation phenotype per se did not limit growth. The additional nitrogen fixation capacity of the rdn1-1 mutant may enhance the benefit of elevated CO2 for plant growth and N2 fixation.

Communities of endophytic bacteria from Cereus peruvianus Mill. (Cactaceae) plants obtained from seeds and from in vitro-regenerated somaclone

Bioprospecting of endophytic microorganisms has been considered a promising resource for obtaining new biomolecules with biotechnological applications. Since few studies are available on the diversity of endophytic bacteria associated with species of the Cactaceae family, current analysis isolates and characterizes the communities of endophytic bacteria associated with the species of C. peruvianus grown in vivo obtained from seed and with somaclone of the same species regenerated in vitro to prospect for their plant-growth promoting activity. Isolation of endophytic bacteria from plants obtained from seeds and from regenerated somaclone in vitro identified nine species of endophytic bacteria (Moraxela osloensis, Corynebacterium mycetoides, Micrococcus luteus, Kocuria palustris, Staphylococcus epidermidis, Staphylococcus cohnii, Pseudomonas stuzeri, Bacillus licheniformis and Microbacterium sp.) not previously described in cacti, and signalized the somaclones as the most promising source of endophytic bacteria for obtaining new biomolecules with biotechnological applications. Somaclones are indicated as the most promising source of endophytic bacteria due to their high concentration of endophytes per gram identified in fresh somaclone tissue, the identification of the highest proportion endophytic bacteria, a source of IAA, and with the highest capacity of biological nitrogen fixation, and EPS production exclusively by the endophytic bacteria isolated from the in vitro regenerated somaclone.

Negative effects of nitrogen override positive effects of phosphorus on grassland legumes worldwide

Anthropogenic nutrient enrichment is driving global biodiversity decline and modifying ecosystem functions. Theory suggests that plant functional types that fix atmospheric nitrogen have a competitive advantage in nitrogen-poor soils, but lose this advantage with increasing nitrogen supply. By contrast, the addition of phosphorus, potassium, and other nutrients may benefit such species in low-nutrient environments by enhancing their nitrogen-fixing capacity. We present a global-scale experiment confirming these predictions for nitrogen-fixing legumes (Fabaceae) across 45 grasslands on six continents. Nitrogen addition reduced legume cover, richness, and biomass, particularly in nitrogen-poor soils, while cover of non-nitrogen-fixing plants increased. The addition of phosphorous, potassium, and other nutrients enhanced legume abundance, but did not mitigate the negative effects of nitrogen addition. Increasing nitrogen supply thus has the potential to decrease the diversity and abundance of grassland legumes worldwide regardless of the availability of other nutrients, with consequences for biodiversity, food webs, ecosystem resilience, and genetic improvement of protein-rich agricultural plant species.

Legume dreams: The contested futures of sustainable plant-based food systems in Europe

With the intensification of agriculture, the simplification of crop rotations, and the rise in demand for meat, dairy and cereal products, legume production and consumption are at an historic low in Europe. But as the environmental consequences of agriculture (biodiversity loss, high greenhouse gas emissions, water pollution) and the health outcomes of modern diets (heart disease, cancer, diabetes, obesity) become better known, so great and varied hopes are being expressed about the future role of legumes in the food system. This paper catalogues and scrutinises these hopes, mapping the promissory narratives now orbiting around legumes. It identifies six food futures, each of which is made possible through the greater use of legumes in various production, processing, marketing and consumption contexts. These promissory narratives are theorised as contrasting responses to three major areas of contestation in the food systems literature. Namely i) the sustainability of livestock management, ii) the role of technology in different visions of the ‘good diet’, and iii) the merits of different models for how to make agricultural management more sustainable. It identifies the promiscuity of legumes – in terms of the range of food futures they permit – before distilling three points of consensus amongst advocates of the potential of legumes. These points of consensus relate to their nitrogen fixing capacity, their high protein content, and their long-standing historical role in the context of European food and farming. This map of legume dreams serves to guide deliberations amongst researchers, policymakers and industry stakeholders about the futures of plant-based food in Europe.

Effects of Nitrogen Application on Nitrogen Fixation Capacity and GmLbs Expression in Soybean

Effects of Nitrogen Application on Nitrogen Fixation Capacity and GmLbs Expression in Soybean

First Report of 16SrII-V Phytoplasma Associated with Green Manure Soybean (Glycine max L.) in Taiwan.

QING PI DOU, a local variety of soybean (Glycine max (L.) Merrill) with small seed size, is primarily cultivated in the southern region of Taiwan. Due to the advantage of high germination rate, fast growth and high nitrogen fixation capacity, QING PI DOU has widely used as green manure in rotation with rice to increase soil fertility in Taiwan. In the summer of 2020, phytoplasma-induced disease symptoms were observed in QING PI DOU with 23% (18/78) disease incidence in Yunlin County, Taiwan. These plants exhibited severe disease symptoms such as little leaf, yellowing, phyllody, virescence, and witches' broom compared to healthy plants. Leaf samples of the symptomatic plants were subsequently collected and examined through transmission electron microscopy (TEM), PCR, and western blotting analyses. The ultrathin sections of the diseased QING PI DOU were double-stained with uranyl acetate and lead citrate. The typical phytoplasma-like pleomorphic bodies were observed in sieve elements of leaf veins by TEM. To investigate the association of phytoplasma with the diseased QING PI DOU, total DNA extracted by the Plant Genomic DNA Purification Kit (DP022, Genemark, Taiwan) was examined by nested PCR using the phytoplasma universal primer pair P1/P7 followed by R16F2n/R16R2 (Lee et al. 1993). The 1.2 kb PCR product specific for 16S ribosomal RNA (16S rRNA) gene was only amplified from symptomatic plants but not from healthy plants. BLAST analysis demonstrated that the sequence (accession no. MW393690) of amplified DNA fragment of 16S rRNA is identical to that of GenBank accession no. NZ_AMWZ01000008 (complement [31109 to 32640]) of peanut witches' broom (PnWB) phytoplasma, a 'Candidatus phytoplasma aurantifolia'-related strain (Firrao et al. 2004). Further analysis on the virtual RFLP pattern of MW393690 generated by iPhyClassifier confirmed that the phytoplasma identified in the diseased QING PI DOU can be classified into the 16SrII-V subgroup. Samples examined by nested PCR were further selected for total cell extracts preparation and characterized by western blotting using the polyclonal antibody raised against the immunodominant membrane protein (Imp) of PnWB phytoplasma (Chien et al. 2020). An expected signal of 19 kDa specific for Imp was only detected in symptomatic plants but not in healthy plants. Moreover, the PCR products encoding SAP11 and phyllogen, the virulence factors responsible for phytoplasma-induced witches' broom and phyllody symptoms (Namba 2019), were also amplified from symptomatic QING PI DOU by PCR using the primer pairs 5'-ATGGCTCCCGAAAAAAATGATAAAGG-3'/5'-TTTTTTAGAATCATCAGGCTTTTTAG-3' (0.28 kb) and 5'-ATGGATCCAAAACTTCCAGAAACT-3'/5'-GTTTTTTTCATCATTTAAATCAT-3' (0.27 kb), respectively. Further analysis by BLAST revealed that SAP11 and phyllogen identified in symptomatic QING PI DOU are identical with those of PnWB phytoplasma. To the best of our knowledge, this report is the first to describe phytoplasma-associated soybean (Glycine max L.) witches' broom disease in green manure soybean in Taiwan.

Molecular Components of Nitrogen Fixation Gene Cluster and Associated Enzymatic Activities of Non-Heterocystous Thermophilic Cyanobacterium Thermoleptolyngbya sp.

Thermoleptolyngbya is a genus of non-heterocystous cyanobacteria that are typical inhabitants of hot spring microbial mats. These filamentous cyanobacteria are capable of nitrogen fixation. In this study, we examined the genome sequences of five publicly available Thermoleptolyngbya strains to explore their nitrogen fixation gene cluster. Analysis of the nitrogen-fixation clusters in these extremophilic strains revealed that the cluster is located in a single locus in Thermoleptolyngbyace. The average nucleotide and amino acid identities of the nitrogen-fixation cluster combined with phylogenetic reconstructions support that nitrogen fixation genes in Thermoleptolyngbyaceae are closely related to one another but also heterogeneous within the genus. The strains from Asia, and China more specifically, generate a separate clade within the genus. Among these strains Thermoleptolyngbya sp. PKUAC-SCTB121 has been selected for experimental validation of clade’s nitrogen fixation capacity. The acetylene reduction experiments of that strain shown that the strain can reduce acetylene to ethylene, indicating a fully functional nitrogenase. The activity of nitrogenase has been tested using different gas compositions across 72 h and exhibited a two-phase trend, high nitrogenase activity at the beginning of the assay that slowed down in the second phase of the analysis.

Microrna Regulation of Nodule Zone-Specific Gene Expression In Soybean

Microrna Regulation of Nodule Zone-Specific Gene Expression In Soybean

BIOTECHNOLOGICAL POTENTIAL OF SOYBEAN PLANT GROWTH-PROMOTING RHIZOBACTERIA

ABSTRACT Technologies that use rhizobacteria to promote plant growth are increasing in agriculture, results have shown improvements in soil quality, increases in productivity, and decreases in the use of synthetic inputs, The objective of work was to characterize bacterial isolates regarding their biological activity and growth promotion of soybean plants grown in a controlled environment. Fifteen bacteria were isolated from soils with continuous use of biological fertilizer. They were evaluated for enzymes production (amylase and protease), nitrogen fixation, antagonistic activity to phytopathogenic fungi, and indoleacetic acid (IAA) production, Soybean seeds were inoculated with bacterial isolates in a greenhouse and evaluated for plant development and soil chemical attributes. The results showed that 8 of the 15 isolates presented production of amylase, protease, or both and 4 isolates presented nitrogen-fixing capacity. The percentage of isolates with high or moderate inhibitory action against the fungi Sclerotinia sclerotiorum, Macrophomina phaseolina, and Fusarium solani were 73.3%, 66.6%, and 73.3%, respectively. The IAA production varied from 8.56 to 31.33 µg mL-1 (5 isolates had low, 6 had moderate, and 4 had high production). The soybean development was significantly higher in 80% of the treatments with inoculation with bacterial isolates. Five bacterial isolates effectively present all characteristics for use as inoculant (biofertilizer) to promote the development of soybean plants.

Exploring the global research trends in biofertilizers: a bibliometric approach.

This research article attempts a bibliometric analysis of global research on biofertilizers carried out from 2000 to 2019. The main purpose of this analysis is in technology foresight; to understand where the research interest lies within the domain of biofertilizer and also to identify the major research networks. The analysis is based on 344 research articles identified using the ISI Web of Science tool, which is processed further using VOSviewer. The results demonstrated that there is an increase in the number of articles, particularly from countries like Brazil, India China, the USA, and Iran. The research focus has been on the assessment of nitrogen fixation capacity of biofertilizers, and the yield improvement due to biofertilizers, and the economics of biofertilizer application. Our findings can act as a useful reference for the researchers, and provide insights for directing future research on biofertilizers.

Genetic Variability of Alnus cordata (Loisel.) Duby Populations and Introgressive Hybridization with A. glutinosa (L.) Gaertn. in Southern Italy: Implication for Conservation and Management of Genetic Resources

Alnus cordata (Loisel.) Duby (Neapolitan alder) is an endemic tree species with a restricted distribution range, limited to Corsica and southern Italy. The economic value of its wood, its rapid growth, the tolerance to drought stress and the nitrogen fixation capacity make A. cordata an excellent candidate for breeding, as well as for conservation and management of genetic resources. In this context, we evaluated the genetic variability of southern Italy populations and verified the hybridization capacity with the simpatric species A. glutinosa (L.) Gaertn. Eight pure A. cordata populations, two pure A. glutinosa populations and six mixed A. cordata/A. glutinosa populations located in southern Italy were analyzed using seven microsatellite markers. A low genetic diversity within and among populations was observed, but no inbreeding effects were evident. A variable frequency of F2 interspecific hybrids was observed in most of the mixed populations and few backcross individuals were scored. These results suggest a limited capacity of hybrid individuals to cross back with the parent species, reducing the risk of genetic pollution of A. cordata. This work provides meaningful knowledge for the conservation and management of the endemic species A. cordata, which represents a valuable source of biodiversity to be conserved.

Regulation of algal and cyanobacterial auxin production, physiology, and application in agriculture: an overview

Algal bioproducts are of growing interest to agriculture because of their biodegradable nature, ability to restore soil fertility, and capacity for plant growth regulation, nitrogen fixation, and carbon sequestration. Plants respond to a suite of growth hormones; auxins present in algal extracts or secreted exogenously by living algae may be partially responsible for the stimulation of plant growth. Auxins are a major class of phytohormones that influence plant growth and development. The roles of auxins in algae and in plants are well described, but studies on the role of auxins in plant-algae interactions remain scarce. This review summarizes the body of knowledge on the production of auxins and their physiological roles in seaweeds, cyanobacteria, and microalgae. Common and differential auxin-associated phenotypes of these algae, including the effect of growth conditions on their auxin production, are also described. Potential mechanisms by which auxins from algae mediate plant development at both phenotypic and molecular levels are also provided. Algal-derived auxins are an environmentally sustainable option for promoting plant growth and yield, but knowledge of their precise mechanisms of action is still rudimentary. Elucidating the pathways by which algal auxins stimulate plant responses and the means by which key environmental factors influence those pathways will help to harness the full potential of algal-derived auxins for agricultural development and resource conservation.

New insights into eutrophication management: Importance of temperature and water residence time

Eutrophication and harmful cyanobacterial blooms threaten water resources all over the world. There is a great controversy about controlling only phosphorus or controlling both nitrogen and phosphorus in the management of lake eutrophication. The primary argument against the dual nutrients control of eutrophication is that nitrogen fixation can compensate the nitrogen deficits. Thus, it is of great necessary to study the factors that can significantly affect the nitrogen fixation. Due to the difference of climate and human influence, the water quality of different lakes (such as water temperature, N:P ratio and water residence time) is also quite different. Numerous studies have reported that the low N:P ratio can intensify the nitrogen fixation capacities. However, the effects of temperature and water residence time on the nitrogen fixation remain unclear. Thus, 30 shallows freshwater lakes in the eastern plain of China were selected to measure dissolved N2 and Ar concentrations through N2: Ar method using a membrane inlet mass spectrometer to quantify the nitrogen fixation capacities and investigate whether the temperature and water residence time have a great impact on nitrogen fixation. The results have shown that the short lake water residence time can severely inhibit the nitrogen fixation capacities through inhibiting the growth of nitrogen-fixing cyanobacteria, changing the N:P ratio and resuspending the solids from sediments. Similarly, lakes with low water temperature also have a low nitrogen fixation capacity, suggesting that controlling nitrogen in such lakes is feasible if the growth of cyanobacteria is limited by nitrogen.

Common Vetch, Valuable Germplasm for Resilient Agriculture: Genetic Characterization and Spanish Core Collection Development.

Common vetch (Vicia sativa L.) is a legume used for animal feed because of its high protein content and great capacity for nitrogen fixation, making this crop relevant in sustainable agriculture. The Spanish vetch collection, conserved at the Spanish Plant Genetic Resources Center (CRF), is one of the largest collections of this species worldwide, including landraces, wild relatives mainly collected in Spain, and commercial cultivars, but also accessions of international origin. The analysis of the genetic diversity of this material, whose genome has not been sequenced yet, and the assembly of a representative collection could play a pivotal role in conserving and exploiting these genetic resources in breeding programs mainly in those focused on consequences and demands of climate change. In this work, a set of 14 simple sequence repeat (SSR) reference alleles for genetic diversity analysis of the CRF vetch collection has been developed, used for genotyping more than 545 common vetch accessions from all over the world and validated. All the tested markers were polymorphic for the analyzed accessions. Overall, at least 86 different loci were identified with 2–11 alleles per locus with an average of 6.1 alleles per locus. Also, the analyses of the generated SSR database support that most of these SSR markers are transferable across closely related species of Vicia genus. Analysis of molecular variance revealed that wild relatives have a higher genetic diversity than landraces. However, cultivars have similar diversity than landraces, indicating that genetic variability has been barely lost due to the breeding of this legume. Low differences of genetic variations between Spanish and non-Spanish accessions have been observed, suggesting a high degree of diversity within Spanish genotypes, which provide 95% of the total genetic variation, so we have focused our efforts on characterizing genotypes of Spanish origin that were further studied using storage protein profiles. Based on SSR, seed protein profiles, and agromorphological and passport data, a vetch core collection (VCC) containing 47 V. sativa accessions of Spanish origin has been established. In this collection, the characterization has been expanded using ISSR markers, and it has been reevaluated with new agromorphological data, including drought tolerance characters. This VCC presents a minimum loss of genetic diversity concerning the total collection and constitutes an invaluable material that can be used in future breeding programs for direct use in a resilient agricultural system.

Vegetative cells may perform nitrogen fixation function under nitrogen deprivation in Anabaena sp. strain PCC 7120 based on genome-wide differential expression analysis.

Nitrogen assimilation is strictly regulated in cyanobacteria. In an inorganic nitrogen-deficient environment, some vegetative cells of the cyanobacterium Anabaena differentiate into heterocysts. We assessed the photosynthesis and nitrogen-fixing capacities of heterocysts and vegetative cells, respectively, at the transcriptome level. RNA extracted from nitrogen-replete vegetative cells (NVs), nitrogen-deprived vegetative cells (NDVs), and nitrogen-deprived heterocysts (NDHs) in Anabaena sp. strain PCC 7120 was evaluated by transcriptome sequencing. Paired comparisons of NVs vs. NDHs, NVs vs. NDVs, and NDVs vs. NDHs revealed 2,044 differentially expressed genes (DEGs). Kyoto Encyclopedia of Genes and Genomes enrichment analysis of the DEGs showed that carbon fixation in photosynthetic organisms and several nitrogen metabolism-related pathways were significantly enriched. Synthesis of Gvp (Gas vesicle synthesis protein gene) in NVs was blocked by nitrogen deprivation, which may cause Anabaena cells to sink and promote nitrogen fixation under anaerobic conditions; in contrast, heterocysts may perform photosynthesis under nitrogen deprivation conditions, whereas the nitrogen fixation capability of vegetative cells was promoted by nitrogen deprivation. Immunofluorescence analysis of nitrogenase iron protein suggested that the nitrogen fixation capability of vegetative cells was promoted by nitrogen deprivation. Our findings provide insight into the molecular mechanisms underlying nitrogen fixation and photosynthesis in vegetative cells and heterocysts at the transcriptome level. This study provides a foundation for further functional verification of heterocyst growth, differentiation, and water bloom control.