Investigation of modulation in P5CS gene expression and some physiological and biochemical traits of wheat in response to Azospirillum and Piriformospora indica inoculation under salinity stress

Document Type : Original Article

Authors

1 Associate Professor, Department of Agronomy and Crop Breeding, Faculty of Agriculture, Ilam University, Ilam, Iran.

2 Ph.D. Graduated in Agrotechnology, Department of Agronomy and Crop Breeding, Faculty of Agriculture, Ilam University, Ilam, Iran.

Abstract

Introduction: Environmental stresses such as salinity are one of the major threats to the sustainability of crop production. Climate change has increased the intensity of these environmental tensions in different regions of the world. Drought and salinity are the critical environmental stresses in arid and semi-arid regions. In recent decades, various strategies have been introduced to reduce the effects of salinity stress on crops. The use of beneficial soil microorganisms is one of these methods.
Materials and methods: In the present study, the effect ofAzospirillum bacteria and Piriformospora indica fungus in separate and combined inoculation was investigated to identify the role of these above-mentioned plant growth-promoting microorganisms in the modulation of the molecular, biochemical, and physiological aspects in wheat under salt stress conditions. A 2×2×2 factorial experimental design was implemented in a completely randomized block design with three replications under greenhouse conditions in 2022. The experiment was conducted in the pot. The main treatments included salt stress imposition, inoculation with and without Azospirillum zaea, and endophytic fungus Piriformospora indica application. Salinity was induced using calcium chloride (65 mg) and sodium chloride (0.375 g) to achieve a salinity level of 12 dS/m. Saline water irrigation was performed 21 and 28 days after wheat germination. Normal water with a salinity of 0.3 decisiemens per meter was used for the control treatment. The wheat cultivar Pishgam was used for this experiment. Twenty-four hours after applying the second salinity treatment, leaf sampling was conducted to extract total RNA and study the expression of the delta-pyrroline-5-carboxylate synthase gene. The changes in leaf proline content and the expression of the gene responsible for proline synthesis (pyrroline-5-carboxylate synthase), malondialdehyde production, ion leakage, leaf relative water content, nitrogen content, and chlorophyll pigment levels in response to applied microorganisms under both salinity and non-salinity conditions were investigated. Additionally, succinate dehydrogenase activity and root colonization rate with fungus were measured under the influence of salinity and inoculation with Azospirillum.
Results: The percentage of fungal colonization in wheat roots of cultivar Pishgam was 24.9%. Inoculation with Azospirillum increased the root colonization rate by 3.17%. Salinity had no significant effect on succinate dehydrogenase, but inoculation with Azospirillum significantly increased succinate dehydrogenase. The plants inoculated with P. indica and Azospirillumhad a higher relative water content in leaves under salt stress. Plants inoculated with bacteria and fungus also had higher nitrogen content. Salinity increased the amount of ion leakage, but inoculation with fungus and bacteria prevented it from increasing. Salinity did not cause a significant change in the amount of chlorophyll pigments, although inoculation with P. indicaand Azospirillum increased the amount of chlorophyll a. The effect of salinity, P. indica, Azospirillum, and their interaction on the expression level of the delta-pyrroline-5 carboxylate synthase gene was significant. Control plants showed higher levels of gene expression, while the inoculated plants with P. indica and Azospirillum had lower gene expression levels. Additionally, the interaction between Azospirillum and P. indica on the expression level of the delta-pyrroline-5 carboxylate synthase gene was significant and it caused a greater decrease compared to the plants that were only inoculated with P. indica. The effect of salinity, P. indica, and bacteria, as well as their mutual effects on leaf proline concentration, was significant. Under salt stress, the amount of proline in the plants inoculated with P. indica was lower than that of the control plants. A similar trend was observed in plants inoculated with bacteria, showing lower levels of proline under salt stress. The expression trend of the delta-pyrroline-5-carboxylate synthase gene was consistent with the trend of proline accumulation under salt stress, indicating the effect of salt induction on the expression of this gene.
Conclusion: The inoculated plants with P. indica and Azospirillum were less affected by induced salt stress than the control plants. The inoculated plants exhibited a lower rate of malondialdehyde and electrolyte leakage, as well as a higher level of chlorophyll pigment and leaf-relative water content. Additionally, P5CS gene expression and proline accumulation in inoculated plants were lower than in control plants.

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References

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Cereal Biotechnology and Biochemistry
Volume 2, Issue 4 - Serial Number 4
December 2024
Pages 391-417

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