Nitrogen Fixation In Legumes Research Articles

Carry‐Over Effect of Leguminous Winter Cover Crops and Living Mulches on Winter Wheat as a Second Main Crop Following White Cabbage

ABSTRACTBackgroundThe direct effect of winter cover crops (WCCs) or living mulches (LMs) on a first vegetable crop has already been investigated. However, little is known about the effect on growth and yield of a second cash crop in the rotation.AimsThe aim of the study was to assess the carry‐over effect of legumes grown as WCC or LM on winter wheat as a second crop after cabbage, measured in yield and nitrogen release.MethodsTwo field trials were carried out in Germany between 2019 and 2022. In the WCC trial, rye, rye with vetch, vetch, pea, and faba bean were used as WCC and compared to bare soil. The WCC biomass was incorporated before cabbage planting in late spring. For the LM trial, perennial ryegrass or white clover was used as LM during cabbage cultivation and compared to bare soil. The LM biomass was incorporated with the cabbage residues and compared to an early incorporation of LM biomass before cabbage planting. In both trials, winter wheat was sown in the fall as the second following main crop in the rotation.ResultsLeguminous WCC species had significant higher wheat yield compared to non‐legumes but not compared to the control without WCC. Late incorporation of LM biomass resulted in increased wheat yield at 10.1–10.4 Mg ha−1 compared to an early incorporation before cabbage planting at 9.35 Mg ha−1. Net N releases show that for WCC, the main effect of legume nitrogen fixation is achieved in the first crop cabbage immediately after incorporation of WCC biomass. In the case of leguminous LM, the effects of legume nitrogen fixation are of much higher relevance in the second main crop, winter wheat, due to LM biomass incorporation after cabbage cultivation.ConclusionTherefore, we suggest to consider not only the direct but also the carry‐over effects of leguminous cover cropping in vegetable crop rotations.

The ‘fertilizer island effect’ cannot eliminate competition between leguminous shrubs and symbiotic herbs in desert ecosystems

The ‘fertilizer island effect’ — a phenomenon engendered by shrubs — promotes the growth of symbiotic herbs and the stabilization of community structures in scrublands. Specifically, leguminous shrubs contribute significantly to the nitrogen input in desert ecosystems. However, the impact of these inputs on the nutrient balance in symbiotic herbs has seldom been explored. To enhance our understanding of the nutrient balance between nitrogen-fixing legume shrubs and symbiotic non-legume herbaceous plants in desert ecosystems, we focused on a typical legume shrub (Eremosparton songoricum) that grows in the Gurbantunggut Desert of northern China. Subsequently, we examined the biomass, ecological stoichiometry, and rhizosphere soil properties of two plant species (annual plant: Ceratocarpus arenarius; and ephemeral plant: Centaurea pulchella) growing across four horizontal distance ranges from E. songoricum. The results indicated that the closer a herbaceous plant was to the leguminous shrubs, the lower was its biomass and nutrient content. In contrast, herbaceous plants growing further away from the shrubs exhibited better growth conditions. The total biomass of Cer. arenarius and Cen. pulchella was greatest at a distance of D4 (90–100 cm), being 3.07, 2.15, and 1.23 times higher for Cer. arenarius and 2.05, 1.18, and 1.26 times higher for Cen. pulchella compared to distances of D1 (0–10 cm), D2 (30–40 cm), and D3 (60–70 cm), respectively. Meanwhile, the physicochemical properties of the rhizosphere root varied inversely with distance, with nutrient levels decreasing horizontally from D1 to D4 and vertically from 0–5 cm > 5–10 cm > 10–20 cm, showcasing a distinct ‘fertilizer island effect’. As the distance increased, the above-ground and below-ground parts of the symbiotic non-leguminous plants started to rely on different types of soil nutrients from different depths to maximize nutrient acquisition. According to the structural equation model, although soil fertility was higher closer to the central leguminous plants, the biomass of nearby herbaceous plants was lower. In summary, in desert ecosystems, the competitive dynamics exerted by leguminous shrubs significantly overshadowed the nutrient provisioning role of the ‘fertilizer island effect’. Furthermore, the utilization of nutrients by symbiotic non-leguminous plants did not exhibit species-specific patterns, and the dominant shrub continued to play a pivotal role. The evidence from this study promotes a deeper understanding of species coexistence mechanisms and ecological stability in desert environments.

EARLY NODULIN93 acts via cytochrome c oxidase to alter respiratory ATP production and root growth in plants.

EARLY NODULIN 93 (ENOD93) has been genetically associated with biological nitrogen fixation in legumes and nitrogen use efficiency in cereals, but its precise function is unknown. We show that hidden Markov models define ENOD93 as a homolog of the N-terminal domain of RESPIRATORY SUPERCOMPLEX FACTOR 2 (RCF2). RCF2 regulates cytochrome oxidase (CIV), influencing the generation of a mitochondrial proton motive force in yeast (Saccharomyces cerevisiae). Knockout of ENOD93 in Arabidopsis (Arabidopsis thaliana) causes a short root phenotype and early flowering. ENOD93 is associated with a protein complex the size of CIV in mitochondria, but neither CIV abundance nor its activity changed in ruptured organelles of enod93. However, a progressive loss of ADP-dependent respiration rate was observed in intact enod93 mitochondria, which could be recovered in complemented lines. Mitochondrial membrane potential was higher in enod93 in a CIV-dependent manner, but ATP synthesis and ADP depletion rates progressively decreased. The respiration rate of whole enod93 seedlings was elevated, and root ADP content was nearly double that in wild type without a change in ATP content. We propose that ENOD93 and HYPOXIA-INDUCED GENE DOMAIN 2 (HIGD2) are the functional equivalent of yeast RCF2 but have remained undiscovered in many eukaryotic lineages because they are encoded by two distinct genes.

Soil application of graphitic carbon nitride nanosheets alleviate cadmium toxicity by altering subcellular distribution, chemical forms of cadmium and improving nitrogen availability in soybean (Glycine max L.)

Cadmium (Cd)-contamination impairs biological nitrogen fixation in legumes (BNF), threatening global food security. Innovative strategies to enhance BNF and improve plant resistance to Cd are therefore crucial. This study investigates the effects of graphitic carbon nitride nanosheets (g-C3N4 NSs) on soybean (Glycine max L.) in Cd contaminated soil, focusing on Cd distribution, chemical forms and nitrogen (N) fixation. Soybean plants were treated with 100 mg kg−1 g-C3N4 NSs, with or without 10 mg kg−1 Cd for 4 weeks. Soil addition of g-C3N4 NSs alleviated Cd toxicity and promote soybean growth via scavenging Cd-mediated oxidative stress and improving photosynthesis. Compared to Cd treatment, g-C3N4 NSs increased shoot and root dry weights under Cd toxicity by 49.5% and 63.4%, respectively. g-C3N4 NSs lowered Cd content by 35.7%–54.1%, redistributed Cd subcellularly by increasing its proportion in the cell wall and decreasing it in soluble fractions and organelles, and converted Cd from high-toxicity to low-toxicity forms. Additionally, g-C3N4 NSs improved the soil N cycle, stimulated nodulation, and increased the N-fixing capacity of nodules, thus increasing N content in shoots and roots by 12.4% and 43.2%, respectively. Mechanistic analysis revealed that g-C3N4 NSs mitigated Cd-induced loss of endogenous nitric oxide in nodules, restoring nodule development. This study highlights the potential of g-C3N4 NSs for remediating Cd-contaminated soil, reducing Cd accumulation, and enhancing plant growth and N fixation, offering new insights into the use of carbon nanomaterials for soil improvement and legume productivity under metal(loid)s stress.

A 1 Ma sedimentary ancient DNA (sedaDNA) record of catchment vegetation changes and the developmental history of tropical Lake Towuti (Sulawesi, Indonesia).

Studying past ecosystems from ancient environmental DNA preserved in lake sediments (sedaDNA) is a rapidly expanding field. This research has mainly involved Holocene sediments from lakes in cool climates, with little known about the suitability of sedaDNA to reconstruct substantially older ecosystems in the warm tropics. Here, we report the successful recovery of chloroplast trnL (UAA) sequences (trnL-P6 loop) from the sedimentary record of Lake Towuti (Sulawesi, Indonesia) to elucidate changes in regional tropical vegetation assemblages during the lake's Late Quaternary paleodepositional history. After the stringent removal of contaminants and sequence artifacts, taxonomic assignment of the remaining genuine trnL-P6 reads showed that native nitrogen-fixing legumes, C3 grasses, and shallow wetland vegetation (Alocasia) were most strongly associated with >1-million-year-old (>1 Ma) peats and silts (114-98.8 m composite depth; mcd), which were deposited in a landscape of active river channels, shallow lakes, and peat-swamps. A statistically significant shift toward partly submerged shoreline vegetation that was likely rooted in anoxic muddy soils (i.e., peatland forest trees and wetland C3 grasses (Oryzaceae) and nutrient-demanding aquatic herbs (presumably Oenanthe javanica)) occurred at 76 mcd (~0.8 Ma), ~0.2 Ma after the transition into a permanent lake. This wetland vegetation was most strongly associated with diatom ooze (46-37 mcd), thought to be deposited during maximum nutrient availability and primary productivity. Herbs (Brassicaceae), trees/shrubs (Fabaceae and Theaceae), and C3 grasses correlated with inorganic parameters, indicating increased drainage of ultramafic sediments and laterite soils from the lakes' catchment, particularly at times of inferred drying. Downcore variability in trnL-P6 from tropical forest trees (Toona), shady ground cover herbs (Zingiberaceae), and tree orchids (Luisia) most strongly correlated with sediments of a predominantly felsic signature considered to be originating from the catchment of the Loeha River draining into Lake Towuti during wetter climate conditions. However, the co-correlation with dry climate-adapted trees (i.e., Castanopsis or Lithocarpus) plus C4 grasses suggests that increased precipitation seasonality also contributed to the increased drainage of felsic Loeha River sediments. This multiproxy approach shows that despite elevated insitu temperatures, tropical lake sediments potentially comprise long-term archives of ancient environmental DNA for reconstructing ecosystems, which warrants further exploration.

Losses of low-germinating, slow-growing species prevent grassland composition recovery from nutrient amendment.

Nutrient enrichment often alters the biomass and species composition of plant communities, but the extent to which these changes are reversible after the cessation of nutrient addition is not well-understood. Our 22-year experiment (15 years for nutrient addition and 7 years for recovery), conducted in an alpine meadow, showed that soil nitrogen concentration and pH recovered rapidly after cessation of nutrient addition. However, this was not accompanied by a full recovery of plant community composition. An incomplete recovery in plant diversity and a directional shift in species composition from grass dominance to forb dominance were observed 7 years after the nutrient addition ended. Strikingy, the historically dominant sedges with low germination rate and slow growth rate and nitrogen-fixing legumes with low germination rate were unable to re-establish after nutrient addition ceased. By contrast, rapid recovery of aboveground biomass was observed after nutrient cessation as the increase in forb biomass only partially compensated for the decline in grass biomass. These results indicate that anthropogenic nutrient input can have long-lasting effects on the structure, but not the soil chemistry and plant biomass, of grassland communities, and that the recovery of soil chemical properties and plant biomass does not necessarily guarantee the restoration of plant community structure. These findings have important implications for the management and recovery of grassland communities, many of which are experiencing alterations in resource input.

Determination of abundance and symbiotic effectiveness of native rhizobia nodulating soybean and other legumes in Rwanda.

Rhizobia diversity in the rhizosphere is one of the key promoters of biological nitrogen fixation between host legumes and microsymbionts, although related complex interaction may depend on various factors. This research was intended to assess the abundance of indigenous rhizobia isolates under various soil conditions, as well as their effectiveness to nodulate legumes such as soybeans. Factors such as soil properties and legume species influence the volume and symbiotic effectiveness of native rhizobia to nodulate crop legumes. To investigate the abundance of rhizobia isolates, legume crops were uprooted to obtain nodules for most probable number (MPN) determination of rhizobia isolates, and soybean (Glycine max.) was used to verify the presence of suitable and efficient rhizobia strains for nitrogen fixation. Soil samples were obtained from the holes out of which nodules were collected, and the laboratory analysis included pH, Mg, K, available P, organic C, Ca, and N to establish the correlation between the soil status and number of rhizobia isolates' cells. Significant variations (p-value <.05) were observed in the cell counts of Rhizobia isolates from Glycine max, Phaseolus vulgaris, Pisum sativum, and Vigna unguiculata, particularly when compared to Arachis hypogaea isolates under acidic conditions. Notably, Pisum sativum and Vigna unguiculata showed consistent performance across all pH conditions. The number of rhizobia isolates was found to be significantly linked to total N and P deficiencies (p < .05). It was also established that total N was dependent on the number of rhizobia cells and that there is a strong correlation between organic carbon and N content. This study highlights the crucial role of understanding and optimizing conditions for rhizobia nodulation in diverse soil environments, emphasizing its potential impact on enhancing biological nitrogen fixation in legumes.

Editorial: Maximizing nitrogen fixation in legumes as a tool for sustainable agriculture intensification, volume II

Editorial: Maximizing nitrogen fixation in legumes as a tool for sustainable agriculture intensification, volume II

Legume nitrogen fixation under nitrate has “FUN” with zinc

Legume nitrogen fixation under nitrate has “FUN” with zinc

An Integrated Affinity Chromatography-Based Approach to Unravel the sRNA Interactome in Nitrogen-Fixing Rhizobia.

The activity mechanism and function of bacterial base-pairing small non-coding RNA regulators (sRNAs) are largely shaped by their main interacting cellular partners, i.e., proteins and mRNAs. We describe here an MS2 affinity chromatography-based procedure adapted to unravel the sRNA interactome in nitrogen-fixing legume endosymbiotic bacteria. The method consists of tagging of the bait sRNA at its 5'-end with the MS2 aptamer followed by pulse overexpression and immobilization of the chimeric transcript from cell lysates by an MS2-MBP fusion protein conjugated to an amylose resin. The sRNA-binding proteins and target mRNAs are further profiled by mass spectrometry and RNAseq, respectively.

Characterization of drought stress-mitigating Rhizobium from faba bean (Vicia faba L.) in the Chinese Qinghai-Tibet Plateau.

Rhizobium-driven symbiotic nitrogen-fixation in legumes not only benefits the growth but also enhances the stress tolerance of plants. Isolating and characterizing efficient, drought-tolerant rhizobia is a central goal for improving crop yields in arid regions. Here, we phylogenetically and biochemically characterized a novel strain of Rhizobium ('QHCD11') sampled from the root nodules of faba beans growing in an arid agricultural area in Qinghai-Tibet. We further tested the drought tolerance of the strain as well as of 'Qingcan 14' faba bean seedlings inoculated with it. Biochemical characterization involved bromothymol blue (BTB) tests, carbon metabolic profiling (Biolog GENIII), DNA-DNA hybridization (dDDH) assays, average nucleotide identity (ANI) analyses, and 16S rRNA sequencing. The result indicated that strain 'QHCD11' likely belongs to the Rhizobium indicum species. Drought stress tolerance was assessed by exposure to polyethylene glycol (PEG-6000) at concentrations of 0, 10, 15, and 20%. Increasing concentrations of PEG-6000 tended to result in decreased growth of 'QHCD11', although the strain performed better at 20% PEG 6000 than at 15%. Inoculation of drought-stressed faba bean seedlings with strain 'QHCD11' improved root vitality, chlorophyll content, antioxidant enzyme activities, and plant height. We suggest that inoculation of faba beans with 'QHCD11' is an environmentally sound strategy for mitigating crop drought stress in arid and semi-arid regions. In addition, the results presents here will benefit future studies into faba bean-rhizobia symbioses under drought stress.

Legume integration for biological control deserves a firmer scientific base

Diversification measures are widely recognized for their potential to enhance the resilience of agri-food production systems under global change. Nitrogen-fixing legumes commonly feature in diversification schemes, where they exert positive impacts on soil structure, nutrient retention, carbon sequestration or fertilizer reduction. Though legumes are premier tools for conservation agriculture, their impacts on (invertebrate, microbial) biological control are far less clear. As legumes secrete copious amounts of energy-rich (floral, extra-floral) nectar and provide alternative host or prey items for resident natural enemies, their deployment is expected to benefit biological control. Yet, its mechanistic basis is poorly understood and scientific underpinnings weak. In this Perspective piece, we collate data from the published literature and open-access databases on the extent to which different natural enemy guilds interact with 25 legume species that are commonly used as intercrops in the (sub)tropics. Our work unveils how natural enemies regularly forage on legumes, but the portrayal of those interaction linkages –or “biostructure” data- is profoundly incomplete and their ensuing implications for biological control are under-investigated. For instance, merely 4 (out of 167) parasitoids of the globally invasive fall armyworm Spodoptera frugiperda have been recorded in association with legumes. We hope that this Perspective piece will stimulate further thought, drive a new wave of biodiversity x ecosystem function research, and ultimately result in the design and implementation (or preservation) of more resilient, resource-frugal and pest-suppressive farming systems.

15 N labeling technology reveals enhancement of nitrogen uptake and transfer by root interaction in cotton/soybean intercropping.

Biological nitrogen fixation in legumes and their transfer of nitrogen to non-legumes in legume/non-legume intercropping systems are considered to be important for the improvement of productivity. However, research on interspecific interaction and root nitrogen transfer in cotton/soybean intercropping systems has rarely been undertaken. In this study, the roots of cotton and soybean were separated with either complete root barriers (CB), using plastic film, or semi-root barriers (SB), using nylon net. No root barrier (NB) was used as the control. The results showed that cotton produced more above-ground dry matter (DM) than soybean. The above-ground DM and nitrogen uptake of cotton was greatest with the NB treatment. The above-ground DM and nitrogen uptake of soybean was greatest with the CB treatment. At the harvest stage, the nitrogen transfer rate from soybean to cotton was 22.47% with the SB treatment and 40.41% with the NB treatment. Interspecific root interaction increased the nitrogen transfer amount, especially for the cotton roots in the 0-15 cm soil layer and for the soybean roots in the 0-30 cm soil layer. The root distribution of soybean was the key factor affecting nitrogen transfer amount, and nitrogen transfer amount was the key factor affecting nitrogen uptake of cotton in the cotton/soybean intercropping system. These results indicated that nitrogen transfer from soybean to cotton through root interaction improved cotton above-ground DM and nitrogen uptake. © 2023 Society of Chemical Industry.

A high-resolution transcriptomic atlas depicting nitrogen fixation and nodule development in soybean.

Although root nodules are essential for biological nitrogen fixation in legumes, the cell types and molecular regulatory mechanisms contributing to nodule development and nitrogen fixation in determinate nodule legumes, such as soybean (Glycine max), remain incompletely understood. Here, we generated a single-nucleus resolution transcriptomic atlas of soybean roots and nodules at 14 days post inoculation (dpi) and annotated 17 major cell types, including six that are specific to nodules. We identified the specific cell types responsible for each step in the ureides synthesis pathway, which enables spatial compartmentalization of biochemical reactions during soybean nitrogen fixation. By utilizing RNA velocity analysis, we reconstructed the differentiation dynamics of soybean nodules, which differs from those of indeterminate nodules in Medicago truncatula. Moreover, we identified several putative regulators of soybean nodulation and two of these genes, GmbHLH93 and GmSCL1, were as-yet uncharacterized in soybean. Overexpression of each gene in soybean hairy root systems validated their respective roles in nodulation. Notably, enrichment for cytokinin-related genes in soybean nodules led to identification of the cytokinin receptor, GmCRE1, as a prominent component of the nodulation pathway. GmCRE1 knockout in soybean resulted in a striking nodule phenotype with decreased nitrogen fixation zone and depletion of leghemoglobins, accompanied by downregulation of nodule-specific gene expression, as well as almost complete abrogation of biological nitrogen fixation. In summary, this study provides a comprehensive perspective of the cellular landscape during soybean nodulation, shedding light on the underlying metabolic and developmental mechanisms of soybean nodule formation.

Rising vapor-pressure deficit increases nitrogen fixation in a legume crop.

Atmospheric vapor-pressure deficit (VPD) is increasing in many regions and has a large impact on plant productivity. A VPD increase leads to raising transpiration rate (TR) and soil-water demand, risking productivity penalties. Like water, nitrogen is critical to productivity, but the effect of VPD on legume nitrogen fixation is undocumented. To address this, we developed a portable system for quantifying nitrogen fixation noninvasively and at a high temporal resolution by tracking the rate of hydrogen gas evolution by root nodules. Combining field and controlled-environment experiments where we measured leaf gas exchange and H2 production by nodules, we confirmed the ability of the system to track nitrogen fixation dynamics. Raising VPD from 0.5 to 3 kPa within c. 2.5 h under well-watered conditions increased nitrogen fixation by up to 25% in addition to TR, consistent with the hypothesis that raising VPD in that range might have alleviated nitrogenase feedback inhibition. Genotypic differences were found in this response, indicating a potential for breeding. Our study provides evidence for an important environmental effect on nitrogen fixation that is not taken into account in current crop and vegetation models, pointing to untapped avenues for better understanding climate change effects on legumes and nitrogen cycling.

Soil Protists Can Actively Redistribute Beneficial Bacteria along Medicago truncatula Roots.

The rhizosphere is the region of soil directly influenced by plant roots. The microbial community in the rhizosphere includes fungi, protists, and bacteria: all play significant roles in plant health. The beneficial bacterium Sinorhizobium meliloti infects growing root hairs on nitrogen-starved leguminous plants. Infection leads to the formation of a root nodule, where S. meliloti converts atmospheric nitrogen to ammonia, a bioavailable form. In soil, S. meliloti is often found in biofilms and travels slowly along the roots, leaving developing root hairs at the growing root tips uninfected. Soil protists are an important component of the rhizosphere system, able to travel quickly along roots and water films, who prey on soil bacteria and have been known to egest undigested phagosomes. We show that a soil protist, Colpoda sp., can transport S. meliloti down Medicago truncatula roots. Using model soil microcosms, we directly observed fluorescently labeled S. meliloti along M. truncatula roots and tracked the displacement of the fluorescence signal over time. Two weeks after co-inoculation, this signal extended 52 mm farther down plant roots when Colpoda sp. was also present versus treatments that contained bacteria but not protists. Direct counts also showed protists are required for viable bacteria to reach the deeper sections of our microcosms. Facilitating bacterial transport may be an important mechanism whereby soil protists promote plant health. IMPORTANCE Soil protists are an important part of the microbial community in the rhizosphere. Plants grown with protists fare better than plants grown without protists. Mechanisms through which protists support plant health include nutrient cycling, alteration of the bacterial community through selective feeding, and consumption of plant pathogens. Here, we provide data in support of an additional mechanism: protists act as transport vehicles for bacteria in soil. We show that protist-facilitated transport can deliver plant-beneficial bacteria to the growing tips of roots that may otherwise be sparsely inhabited with bacteria originating from a seed-associated inoculum. By co-inoculating Medicago truncatula roots with both S. meliloti, a nitrogen-fixing legume symbiont, and Colpoda sp., a ciliated protist, we show substantial and statistically significant transport with depth and breadth of bacteria-associated fluorescence as well as transport of viable bacteria. Co-inoculation with shelf-stable encysted soil protists may be employed as a sustainable agriculture biotechnology to better distribute beneficial bacteria and enhance the performance of inoculants.

The putative transporter MtUMAMIT14 participates in nodule formation in Medicago truncatula

Transport systems are crucial in many plant processes, including plant–microbe interactions. Nodule formation and function in legumes involve the expression and regulation of multiple transport proteins, and many are still uncharacterized, particularly for nitrogen transport. Amino acids originating from the nitrogen-fixing process are an essential form of nitrogen for legumes. This work evaluates the role of MtN21 (henceforth MtUMAMIT14), a putative transport system from the MtN21/EamA-like/UMAMIT family, in nodule formation and nitrogen fixation in Medicago truncatula. To dissect this transporter’s role, we assessed the expression of MtUMAMIT14 using GUS staining, localized the corresponding protein in M. truncatula root and tobacco leaf cells, and investigated two independent MtUMAMIT14 mutant lines. Our results indicate that MtUMAMIT14 is localized in endosomal structures and is expressed in both the infection zone and interzone of nodules. Comparison of mutant and wild-type M. truncatula indicates MtUMAMIT14, the expression of which is dependent on the presence of NIN, DNF1, and DNF2, plays a role in nodule formation and nitrogen-fixation. While the function of the transporter is still unclear, our results connect root nodule nitrogen fixation in legumes with the UMAMIT family.

Widely conserved AHL transcription factors are essential for NCR gene expression and nodule development in Medicago

Symbiotic nitrogen fixation by Rhizobium bacteria in the cells of legume root nodules alleviates the need for nitrogen fertilizers. Nitrogen fixation requires the endosymbionts to differentiate into bacteroids which can be reversible or terminal. The latter is controlled by the plant, it is more beneficial and has evolved in multiple clades of the Leguminosae family. The plant effectors of terminal differentiation in inverted repeat-lacking clade legumes (IRLC) are nodule-specific cysteine-rich (NCR) peptides, which are absent in legumes such as soybean where there is no terminal differentiation of rhizobia. It was assumed that NCRs co-evolved with specific transcription factors, but our work demonstrates that expression of NCR genes does not require NCR-specific transcription factors. Introduction of the Medicago truncatula NCR169 gene under its own promoter into soybean roots resulted in its nodule-specific expression, leading to bacteroid changes associated with terminal differentiation. We identified two AT-Hook Motif Nuclear Localized (AHL) transcription factors from both M. truncatula and soybean nodules that bound to AT-rich sequences in the NCR169 promoter inducing its expression. Whereas mutation of NCR169 arrested bacteroid development at a late stage, the absence of MtAHL1 or MtAHL2 completely blocked bacteroid differentiation indicating that they also regulate other NCR genes required for the development of nitrogen-fixing nodules. Regulation of NCRs by orthologous transcription factors in non-IRLC legumes opens up the possibility of increasing the efficiency of nitrogen fixation in legumes lacking NCRs.

Trends in the direction of global plant invasion biology research over the past two decades.

Invasive plants are a growing ecological problem worldwide, but biases and patterns within invasive plant research may affect our understanding of invasive plant ecology. In this study, we analyzed 458 invasive plant papers sampled from the two journals dedicated entirely to the field of invasion biology, i.e., Biological Invasions and Neobiota. From these papers, we collected information on geographic coverage, climate, habitat, taxonomic coverage, plant functional type, and research topic to examine trends across a 21-year time period from 1999 to 2020. Our analysis found that invasive plant research was consistently biased toward temperate grassland and forest ecosystems particularly within the Americas, Europe, and Australia, and toward smaller, herbaceous invasive plant species (i.e., forbs, grasses, and shrubs), with an increase in interest in invasive nitrogen-fixing legumes over time. Our analysis also identified "hot" research topics in invasive plant research at specific time periods, such as a peak in the use of genetic analysis methods in 2014-2015 and a more recent focus on plant physiological and functional traits. While current models, concepts, and understanding of plant invasion ecology are still driven by such biases, this has been partially offset by recent increased research in understudied systems, as well as increasing awareness that plant invasion is heavily affected by their growth types, physiological traits, and soil interactions. As the field of invasion biology becomes ever increasingly important over time, focusing invasive plant research on understudied ecosystems and plant groups will allow us to develop a more holistic understanding of the ecology of invasive plants. In particular, given the outsized importance of the tropics to global biodiversity, the threats they face, and the dearth of studies, it is of critical importance that more invasive plant research is conducted within the tropics to develop a more globally representative understanding of invasive plant ecology.

PHYTOCHEMICAL SCREENING AND ANTIOXIDANT VITAMINS OF ACACIA NILOTICA POD

Acacia nilotica is also known as Gum Arabic tree, Babul, Egyptian thorn, or Prickly Acacia is multipurpose nitrogen fixing legume tree. The aim of this work was to determine the phytochemical constituent and antioxidants vitamins of Acacia nilotica pod. The Acacia nilotica were obtained from Dange Shuni and the pod was also obtained by removing the seeds. They were properly screened and identified by Botanist in the herbarium of botany unit, biological science, Usmanu Danfodiyo University, Sokoto. 5g of the sample with 100ml of distilled waterwas soaked overnight and filtered to obtain the filtrate (extract). The samples were analyzed using spectrophotometer. Phytochemical analysis showed the presence of Alkaloids, Flavonoids, Saponins, Tannins, Cardiac glycosides, Steroids, Glycosides, Balsam and Volatile oils. However, anthraquinones were not detected. The quantitative result showed Alkaloids to be (15.9 ± 0.31), Flavonoids (3.8 ± 1.22), Tannins (0.1077 ± 0.00058), Steroids (0.073 ± 0.00038) and Glycosides (5.77 ± 0.015). The antioxidant vitamins analyzed showed vitamin C concentration to be (2.3018±0.0017), vitamin E (329.9 ± 0.20) and vitamin A equivalent (84.72 ± 0.39). This shows that A. nilotica pod is rich in Phytochemicals and antioxidant vitamins.