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1.
Bio Protoc ; 13(20): e4855, 2023 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-37965269

RESUMO

Maize is one of the most important crops in the world, and ensuring its successful growth and productivity is crucial for global food security. One way to enhance maize growth and productivity is by improving the colonization of its roots by beneficial microorganisms. In this regard, Serendipita indica, a plant growth-promoting fungus, has gained attention for its ability to enhance plant growth and productivity, especially in cereal crops and medicinal plants. Previous studies have shown that S. indica can colonize various plant species, including maize, but the efficiency of the colonization process in maize seedlings has not been extensively characterized. This protocol outlines a method for efficient colonization of maize seedlings with the beneficial fungus S. indica. The protocol includes the preparation of stock solutions, maintenance and growth of S. indica, surface sterilization and germination of seeds, preparation of S. indica chlamydospores, and colonization of maize plants with S. indica. The advantages of this protocol include the use of surface sterilization techniques that minimize contamination, the production of a large number of viable chlamydospores, and efficient colonization of maize seedlings with S. indica. This protocol may be useful for researchers studying the role of S. indica in promoting plant growth and combating biotic and abiotic stress. Additionally, this protocol may be used in the development of biofertilizers using S. indica as a means of increasing crop yields and reducing dependence on synthetic fertilizers. Overall, this protocol offers a reliable and efficient method for colonizing maize seedlings with S. indica and may have potential applications in the agricultural industry. This study also provides a valuable tool for researchers interested in studying plant-microbe interactions in maize and highlights the potential of S. indica as a biocontrol agent to enhance maize productivity under adverse conditions. Key features • This protocol builds upon the method developed by Narayan et al. (2022), and its application optimized for the root endophytic symbiotic fungus S. indica. • This protocol also allows for histochemical analysis to visualize the colonized fungal spores in the root cells of host plant species. • This protocol helps in mathematical calculation of the percent colonization or efficiency of colonization. • This protocol utilizes readily available laboratory equipment, including a light microscope, autoclave, and laminar flow hood, ensuring ease of reproducibility in other research laboratories.

2.
Environ Pollut ; 290: 117953, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34438168

RESUMO

Hydrogen sulfide (H2S) has emerged as a potential gasotransmitter in plants with a beneficial role in stress amelioration. Despite the various known functions of H2S in plants, not much information is available to explain the associative role of molybdenum (Mo) and hydrogen sulfide (H2S) signaling in plants under arsenic toxicity. In view to address such lacunae in our understanding of the integrative roles of these biomolecules, the present work attempts to decipher the roles of Mo and H2S in mitigation of arsenate (AsV) toxicity in faba bean (Vicia faba L.) seedlings. AsV-stressed seedlings supplemented with exogenous Mo and/or NaHS treatments (H2S donor) showed resilience to AsV toxicity manifested by reduction of apoptosis, reactive oxygen species (ROS) content, down-regulation of NADPH oxidase and GOase activity followed by upregulation of antioxidative enzymes in leaves. Fluorescent localization of ROS in roots reveals changes in its intensity and spatial distribution in response to MO and NaHS supplementation during AsV stress. Under AsV toxicity conditions, seedlings subjected to Mo + NaHS showed an increased rate of nitrogen metabolism evident by elevation in nitrate reductase, nitrite reductase and glutamine synthetase activity. Furthermore, the application of Mo and NaHS in combination positively upregulates cysteine and hydrogen sulfide biosynthesis in the absence and presence of AsV stress. Mo plus NaHS-supplemented seedlings exposed to AsV toxicity showed a substantial reduction in oxidative stress manifested by reduced ELKG, lowered MDA content and higher accumulation of proline in leaves. Taken together, the present findings provide substantial evidence on the synergetic role of Mo and H2S in mitigating AsV stress in faba bean seedlings. Thus, the application of Mo and NaHS reveals their agronomic importance to encounter heavy metal stress for management of various food crops.


Assuntos
Arsênio , Sulfeto de Hidrogênio , Vicia faba , Arsênio/toxicidade , Cisteína , Molibdênio/toxicidade , Nitrogênio , Plântula
3.
Biochem Biophys Res Commun ; 394(1): 163-9, 2010 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-20188073

RESUMO

This study presents first hand data on the cloning and heterologous expression of Anabaena PCC 7120 all3940 (a dps family gene) in combating nutrients limitation and multiple abiotic stresses. The Escherichia coli transformed with pGEX-5X-2-all3940 construct when subjected to iron, carbon, nitrogen, phosphorus limitation and carbofuron, copper, UV-B, heat, salt and cadmium stress registered significant increase in growth over the cells transformed with empty vector under iron (0%), carbon (0.05%), nitrogen (3.7 mM) and phosphorus (2mM) limitation and carbofuron (0.025 mg ml(-1)), CuCl(2) (1 mM), UV-B (10 min), heat (47 degrees C), NaCl (6% w/v) and CdCl(2) (4mM) stress. Enhanced expression of all3940 gene measured by semi-quantitative RT-PCR at different time points under above mentioned treatments clearly demonstrates its role in tolerance against aforesaid abiotic stresses. This study opens the gate for developing transgenic cyanobacteria capable of growing successfully under above mentioned stresses.


Assuntos
Anabaena/genética , Proteínas de Bactérias/genética , Proteínas de Ligação a DNA/genética , Estresse Fisiológico/genética , Cloreto de Cádmio/toxicidade , Carbono/deficiência , Clonagem Molecular , Escherichia coli/efeitos dos fármacos , Escherichia coli/genética , Escherichia coli/efeitos da radiação , Expressão Gênica , Temperatura Alta , Deficiências de Ferro , Nitrogênio/deficiência , Fósforo/deficiência , Raios Ultravioleta
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