Recent progress in the role of seed endophytic bacteria as plant growth-promoting microorganisms and biocontrol agents.

The importance of microorganisms residing within the host plant for their growth and health is increasingly acknowledged, yet the significance of microbes associated with seeds, particularly seed endophytic bacteria, remains underestimated. Seeds harbor a wide range of bacteria that can boost the growth and resilience of their host plants against environmental challenges. These endophytic associations also offer advantages for germination and seedling establishment, as seed endophytic bacteria are present during the initial stages of plant growth and development. Furthermore, plants can selectively choose bacteria possessing beneficial traits, which are subsequently transmitted through seeds to confer benefits to future generations. Interestingly, even with the ongoing discovery of endophytes in seeds through high-throughput sequencing methods, certain endophytes remain challenging to isolate and culture from seeds, despite their high abundance. These challenges pose difficulties in studying seed endophytes, making many of their effects on plants unclear. In this article, a framework for understanding the assembly and function of seed endophytes, including their sources and colonization processes was outlined in detail and available research on bacterial endophytes discovered within the seeds of various plant species has also been explored. Thus, this current review aims to provide valuable insights into the mechanism of underlying seed endophytic bacteria-host plant interactions and offers significant recommendations for utilizing the seed endophytic bacteria in sustainable agriculture as plant growth promoters and enhancers of environmental stress tolerance.

Novel report of Acinetobacter johnsonii as an indole-producing seed endophyte in Tamarindus indica L.

Plant-microbe associations have been regarded as an exciting topic of research due to their potential as environment friendly alternatives for stimulating crop growth and development. Seeds of Tamarindus indica L. have been chosen for the present study as seed endophytes prefer larger or nutritive cotyledon and hard seed coats for their colonization. The main objectives of our study were to isolate and identify the seed endophytes, their bioefficacy, and responsible chemical compounds. In a dose-dependent experiment, tamarind seed exudates (TSE) showed plant growth-promoting properties on Oryza sativa (53-81%), Daucus carota (10-31%), and Raphanus sativa (21-42%). Identification of the bacterial load in TSE through 16S rRNA sequencing revealed the existence of two bacterial species, Acinetobacter johnsonii and Niallia nealsonii. This is the first report of these two bacteria as seed endophytes of Tamarindus indica L. HRLC-MS analysis of TSE confirmed the presence of indole derivatives, primarily indole-3-lactic acid (ILA). The quantitative phytochemical estimation of bacterial culture filtrates revealed that indole-like substances were present in the extracts only in A. johnsonii at a concentration of 0.005 mg/ml of indole acetic acid equivalent. Experimental results suggested that the stimulatory activity of TSE was caused by the presence of A. johnsonii, a potential plant growth-promoting bacteria that produced indole-like compounds. This study suggests tamarind seed exudates with its endophytic microbiota as a potent plant growth-promoting agent that may find use as a cheap and sustainable source of metabolites useful in the agro-industries.

Surface functionalization and size of polystyrene microplastics concomitantly regulate growth, photosynthesis and anti-oxidant status of Cicer arietinum L.

Microplastics are a recent entrant in the list of environmental pollutants, exhibiting great diversity owing to different sizes, surface charges, and morphologies. The present study explores the impact of varied size, surface functionalization, and concentration of polystyrene microplastics (PS MP) on plants. For this study, Cicer seedlings were exposed to two different sizes of PS (1 µm and 12 µm) with three different surface functionalization (plain, carboxylated, and aminated) and at three distinct concentrations (10, 50, and 100 mg/L). The growth and photosynthetic parameters (like pigment content, Hill activity, etc.) along with oxidative stress marker (ROS) and anti-oxidant enzyme activities (like Superoxide dismutase, Catalase, and Peroxidase) were assessed. The results incline towards the idea that with increasing concentration of PS, there was a decline in the growth of the seedlings. There was also a dose-dependent increase in oxidative stress due to the suppression of the action of antioxidant enzymes. The effect was more prominent for 12 µm PS, perhaps due to its larger size and adherence to roots resulting in mechanical damage as deduced from MDA levels in the seedlings. Besides, MP with negative surface charge was comparatively less toxic than uncharged or positively charged PS of 1 µm. Overall, it can be concluded that the impact of MP on plants does not rely on individual characteristics of the particles alone, rather it is a concerted result of various determinants like size, charge, and concentration.

Nano-Scale Zero Valent Iron (nZVI) Priming Enhances Yield, Alters Mineral Distribution and Grain Nutrient Content of Oryza sativa L. cv. Gobindobhog: A Field Study.

In recent decades, nano-scale zero valent iron is reported to have plant growth enhancement capacity under laboratory conditions, but till date, there is no report to highlight its effect on the growth and yield of field-grown plants. In this study, we have evaluated the potential of nZVI priming on rice yield. A two-year field study has been conducted with different concentrations (10, 20, 40, and 80 mg l-1) of nZVI for seed priming. The efficacy of nanopriming was compared with the hydroprimed control set. Seeds were treated for 72 h and sown in nursery beds and after 30 days seedlings were transplanted in the field. Root anatomy and morphology were studied in 7 days old seedlings where no changes were found. RAPD analysis also confirmed that low doses of nZVI were not genotoxic. Nanoprimed plants also had broader leaves, higher growth, biomass, and tiller number than control plants. Maximum yield was obtained from the 20 mg l-1 nZVI primed set (3.8 fold higher than untreated control) which is achieved primarily because of the increase in fertile tiller numbers (two fold higher than untreated control). Higher values of other agronomic parameters like growth rate, net assimilation rate proved that nZVI priming enhanced photosynthetic efficiency and helped in the proper storage of photo-assimilates. All these attributed to increased accumulation of phytochemicals like starch, soluble sugar, protein, lipid, phenol, riboflavin, thiamine, and ascorbic acid in the grains. The elemental analysis confirmed that nZVI priming also promoted higher accumulations of macro and micronutrients in grains. Thus, nanoprimed seeds showed better crop performance compared to the traditional hydropimed seeds. Hence, nZVI can be considered as 'pro-fertilizer' and can be used commercially as a seed treatment agent which is capable of boosting plant growth and yield along with minimum interference to the soil ecosystem. Supplementary Information: The online version contains supplementary material available at 10.1007/s00344-021-10335-0.

Fe3O4-urea nanocomposites as a novel nitrogen fertilizer for improving nutrient utilization efficiency and reducing environmental pollution.

Almost 81% of nitrogen fertilizers are applied in form of urea but most of it is lost due to volatilization and leaching leading to environmental pollution. In this regard, slow-release nano fertilizers can be an effective solution. Here, we have synthesized different Fe3O4-urea nanocomposites with Fe3O4 NPs: urea ratio (1:1, 1:2, 1:3) ie. NC-1, 2, and 3 respectively, and checked their efficacy for growth and yield enhancement. Oryza sativa L. cv. Swarna seedlings were treated with different NCs for 14 days in hydroponic conditions and significant up-regulation of photosynthetic efficiency and nitrogen metabolism were observed due to increased availability of nitrogen and iron. The discriminant functional analysis confirmed that the NC3 treatment yielded the best results so further gene expression studies were performed for NC-3 treated seedlings. Significant changes in expression profiles of ammonia and nitrate transporters indicated that NC-3 treatment enhanced nitrogen utilization efficiency (NUE) due to sustained slow release of urea. From pot experiments, we found significant enhancement of growth, grain nutrient content, and NUE in NC supplemented sets. 1.45 fold increase in crop yield was achieved when 50% N was supplemented in form of NC-3 and the rest in form of ammonium nitrate. NC supplementation can also play a vital role in minimizing the use of bulk N fertilizers because, when 75% of the recommended N dose was supplied in form of NC-3, 1.18 fold yield enhancement was found. Thus our results highlight that, slow-release NC-3 can play a major role in increasing the NUE of rice.

Elucidating ROS signaling networks and physiological changes involved in nanoscale zero valent iron primed rice seed germination sensu stricto.

Reactive oxygen species (ROS) play pivotal roles during seed dormancy and germination. Metabolically active cells of seeds generate ROS and successful germination is governed by internal ROS contents, maintained within an optimum "oxidative window" by several ROS scavengers. Although ROS was previously considered hazardous, optimum ROS generation in seeds can mediate early seed germination by acting as messengers for cell signaling involved in endosperm weakening, stored food mobilization, etc. Recent reports suggest that nanopriming can expedite seed germination rates and enhance seed quality and crop performances. However, nanoparticle-driven signal cascades involved during seed germination are still unknown. The present study is aimed to explore molecular mechanisms for promoting germination in nanoprimed seeds and to investigate the plausible role of nanoparticle-mediated ROS generation in this process. Here rice seeds were primed with 20 mg L -1nanoscale zero valent iron (nZVI) for 72 h and several biochemical and physiological changes were monitored at different time points (5, 10, 20, 40, 60, and 80 h). To gain insight into roles of ROS in germination rate enhancement, intercellular ROS inhibitor, diphenyleneiodonium (DPI) was taken as another priming agent. Seed priming with DPI impaired seed germination percentage, hydrolytic enzyme activities due to ROS imbalance. On the contrary, seeds primed with both DPI and nZVI could recover from deleterious consequences of DPI treatment. Although DPI impaired intercellular ROS generation, nZVI can generate ROS independently which was confirmed from ROS localization assay. In both nZVI and the DPI and nZVI co-primed sets, significant up-regulation in genes like OsGA3Ox2, OsGAMYB were observed which are responsible for regulating the activity of several hydrolases and mediates efficient mobilization of storage food reserves of seeds. Thus, nZVI priming has potential to regulate intracellular ROS levels and orchestrate all the metabolic activities which eventually up-regulates seed germination rate and seed vigour.

Nanopriming with zero-valent iron synthesized using pomegranate peel waste: A "green" approach for yield enhancement in Oryza sativa L. cv. Gonindobhog.

Nanopriming is a combination of nanoparticle treatment and a seed dressing technique that can increase seed quality, seedling vigour, yield and also imparts tolerance against biotic and abiotic stress. Here, nano-scale zero-valent iron (G-nZVI) was synthesized using fruit peel waste of Punica granatum L and their formation was validated from XRD and optical spectroscopic techniques. Later, the seeds were primed with G-nZVI at six different concentrations (0, 10, 20, 40, 80, and 160 mg L -1) to determine the dose which is optimum for increasing germination percentage and seedling vigour of rice (Oryza sativa L. cv. Gobindobhog). According to initial results, upon priming seeds with 40-80 mg L -1 G-nZVI highest growth rate was found. The early growth enhancement of seedlings was chiefly attributed to increased ROS generation, higher hydrolytic enzyme activities, and increased iron uptake in germinating seeds upon nanopriming. The effects of nanopriming were carried over to later stages of development. A field experiment was carried out where nanoprimed seeds and traditional hydroprimed control seeds were sown in plots and grown till maturity without the aid of any conventional fertilizers and pesticides and it was found that crop yield and grain nutrient concentrations were higher in nanoprimed sets. Compared to control hydroprimed sets, 1.53 folds higher crop yield was observed upon seed priming with 80 mg L -1 G-nZVI. Thus in the future, G-nZVI can be considered to be a novel low-cost, eco-friendly, food waste-derived seed treatment agent that has immense potential in increasing rice yield.

Differential growth and metabolic responses induced by nano-scale zero valent iron in germinating seeds and seedlings of Oryza sativa L. cv. Swarna.

Since development of antioxidant defence system is high energy demanding event, innate defence system and stress tolerance of plant is strictly governed by plant age. This study is aimed towards evaluating variation of tolerance in germinating seeds and seedlings of Oryza sativa L. cv. Swarna against nano-scale zero valent iron (nZVI). A comparative study of several physiological and biochemical parameters have been carried out among 2 distinct plant groups, Group I treated with variable concentrations of nZVI (50, 100, 150 and 200 mg L-1) during germination and Group II treated with similar nZVI doses on 7th day after germination. Upon treatment with higher nZVI concentrations, Group I seedlings showed susceptibility towards oxidative stress while Group II seedlings showed tolerance against these higher doses of nZVI. Significant growth enhancement was observed upon treatment with 50-150 mg L-1 nZVI, since up-regulation of plant's endogenous antioxidant system protected relatively aged Group II seedlings from oxidative damages. Hierarchical clustering based on overall physiological, biochemical and stress parameters confirmed that in Group I seedlings 100-200 mg L-1 nZVI treatments were toxic where as in Group II seedlings 50-150 mg L-1 nZVI treatments showed growth promoting effects. This differential response is due to developmental stage related resistance in plants.

Nano-scale zero valent iron modulates Fe/Cd transporters and immobilizes soil Cd for production of Cd free rice.

In recent decades, nanoscale zero valent iron (nZVI) has been found to be a promising approach for heavy metal remediation. This study is the first report highlighting the role of nZVI to ameliorate Cadmium (Cd) stress in rice along with its effects in expressions of transporter genes, agronomic parameters and grain nutrient status. Initially, 3 concentration of Cd (10, 50, 250 µM) and nZVI (50, 100, 200 mg L-1) were selected. PCA analysis based on growth parameters, photosynthetic pigment contents and lipid peroxidation rate confirmed that 100 mg L-1 nZVI was most suitable for remediation of 10 µM Cd. It was evident that, nZVI can alleviate Cd-induced toxic effects by enhancing antioxidant defense mechanisms and other physiological processes in plants. nZVI treated rice seedlings also showed upregulation of phytochelatins which aided in Cd chelation within vacuoles. Study of root morphology with scanning electron microscopy and ROS imaging with confocal microscopy confirmed that nZVI could alleviate oxidative stress due to Cd uptake. In nZVI treated rice seedlings, gene expressions of iron (Fe) transporters (like, IRT1,IRT2,YSL2,YSL15) which are responsible for both Fe and Cd uptake were significantly down-regulated whereas, OsVIT1 and OsCAX4 genes were over expressed which lead to sequestration of Cd in vacuoles. Cd localization assay with leadmium proved that Cd translocation was reduced with nZVI treatment. To further validate our findings a pot experiment was carried out where it was found that nZVI could immobilize Cd in soil prevented accumulation of Cd in rice grains in addition to improving yield.

Nanocomposites for Delivering Agrochemicals: A Comprehensive Review.

Excessive application of fertilizers negatively affects soil health, causes low nutrient utilization efficiency in plants, and leads to environmental pollution. The application of controlled-release fertilizer is gaining momentum to overcome this crisis. Engineered nanocomposites (ENCs) have shown tremendous promise for need-based delivery of agrochemicals (macro- and micronutrients, pesticides, and other agrochemicals). This review provides comprehensive coverage of synthesis of nanocomposites, their physical-chemical characterization, and techniques to achieve sustained release and targeted delivery to the crops, emphasizing their beneficial role in plant production and protection. Related aspects like feasibility of the application, commercialization of the nanoformulations, and biosafety concerns are also highlighted. This will be helpful to develop a critical understanding of the current state of the art in the controlled release of agrochemicals through nanocomposites. The pressing issues like scale up production, cost analyses, field-based trials, and environmental safety concerns should be given greater attention in future studies.

Nanopriming with zero valent iron (nZVI) enhances germination and growth in aromatic rice cultivar (Oryza sativa cv. Gobindabhog L.).

Engineered nanoparticles are utilized in agriculture for various purposes. They can be used as fertilizer, carrier for macro/micro nutrients or priming agents. Various nanoparticles are reported to have toxicity at very high doses, but at optimum concentration, they can be beneficial for plant growth and development. In the present study, low concentrations of nZVI nanoparticles were evaluated for their growth enhancement potential as seed priming agent in an aromatic rice cultivar, Oryza sativa cv. Gobindabhog. Seeds were primed with different concentrations (10, 20, 40, 80, 160 mg L-1) of nZVI and allowed to grow for 14 days. Seed germination and seedling growth were studied by assessing physiological, biochemical, and structural parameters at different time points. Maximum activities of hydrolytic and antioxidant enzymes, along with root dehydrogenase enzyme were observed in 20 mg L-1 nZVI primed seeds. Priming with low doses of nZVI increased seedling vigour, as expressed by increased root and shoot length, biomass and photosynthetic pigment content. Our study also confirmed that after 14 days growth, the seedling showed absence of membrane damage, reduction in proline level and anti-oxidant enzyme activities. However, seedlings primed with 160 mg L-1 nZVI suffered oxidative stress. SEM micrographs also revealed damage in root tissue at that concentration. AAS study confirmed uptake of nZVI by the rice plants as maximum level of iron was found in the plants treated with highest concentration (i.e. 160 mg L-1 nZVI). Thus, nZVI at low concentrations can be considered as priming agent of rice seeds for increasing plant vigour.