Co-inoculation of mycorrhizal fungi and plant growth-promoting rhizobacteria improve growth, biochemical and physiological attributes in Dracocephalum kotschyi Boiss. under water deficit stress.

Background: Because of swift climate change, drought is a primary environmental factor that substantially diminishes plant productivity. Furthermore, the increased use of chemical fertilizers has given rise to numerous environmental problems and health risks. Presently, there is a transition towards biofertilizers to enhance crops' yield, encompassing medicinal and aromatic varieties. Methods: This study aimed to explore the impacts of plant growth-promoting rhizobacteria (PGPR), both independently and in conjunction with arbuscular mycorrhizal fungi (AMF), on various morphological, physiological, and phytochemical characteristics of Dracocephalum kotschyi Boiss. This experimentation took place under different irrigation conditions. The irrigation schemes encompassed well watering (WW), mild water stress (MWS), and severe water stress (SWS). The study evaluated the effects of various biofertilizers, including AMF, PGPR, and the combined application of both AMF and PGPR (AMF + PGPR), compared to a control group where no biofertilizers were applied. Results: The findings of the study revealed that under water-stress conditions, the dry yield and relative water content of D. kotschyi Boiss. experienced a decline. However, the application of AMF, PGPR, and AMF + PGPR led to an enhancement in dry yield and relative water content compared to the control group. Among the treatments, the co-application of AMF and PGPR in plants subjected to well watering (WW) exhibited the tallest growth (65 cm), the highest leaf count (187), and the most elevated chlorophyll a (0.59 mg g-1 fw) and b (0.24 mg g-1 fw) content. Regarding essential oil production, the maximum content (1.29%) and yield (0.13 g plant -1) were obtained from mild water stress (MWS) treatment. The co-application of AMF and PGPR resulted in the highest essential oil content and yield (1.31% and 0.15 g plant-1, respectively). The analysis of D. kotschyi Boiss. essential oil identified twenty-six compounds, with major constituents including geranyl acetate (11.4-18.88%), alpha-pinene (9.33-15.08%), Bis (2-Ethylhexyl) phthalate (8.43-12.8%), neral (6.80-9.32%), geranial (9.23-11.91%), and limonene (5.56-9.12%). Notably, the highest content of geranyl acetate, geranial, limonene, and alpha-pinene was observed in plants subjected to MWS treatment following AMF + PGPR application. Furthermore, the co-application of AMF, PGPR, and severe water stress (SWS) notably increased the total soluble sugar (TSS) and proline content. In conclusion, the results indicate that the combined application of AMF and PGPR can effectively enhance the quantity and quality of essential oil in D. kotschyi Boiss., particularly when the plants are exposed to water deficit stress conditions.

Comprehensive proteomic analysis of canola leaf inoculated with a plant growth-promoting bacterium, Pseudomonas fluorescens, under salt stress.

Plant growth-promoting bacteria can improve the tolerance of canola to salt stress. To better understand the effects of plant growth-promoting bacterium on the protein profiles of canola under salt stress condition, proteomics was performed. Salt-sensitive (Sarigol) and -tolerant (Hyola308) canola cultivars were inoculated with Pseudomonas fluorescens FY32, and the protein profiles of canola leaves were compared using a PEG-fractionation method. Cluster analysis of canola cultivars based on a stress tolerance index of several morphological parameters was used to confirm that Sarigol and Hyola308 were salt-sensitive and -tolerant cultivars, respectively. Using a gel-free proteomic technique, 154 and 94 proteins in Hyola308 and 100 and 144 proteins in Sarigol were uniquely identified in non-inoculated and bacterial-inoculated cultivars, respectively. By PEG fractionation, a total of 132 and 207 proteins were identified in non-inoculated and inoculated Hyola308, respectively. Notably, the abundance of copper/zinc superoxide dismutase 1 was significantly increased in inoculated Hyola308 under severe salt stress and decreased under moderate salt stress. In addition, the enzyme activity of delta-1-pyrroline-5-carboxylate synthase was significantly increased non-inoculated Hyola308 and the activity of succinate dehydrogenase was increased in inoculated Hyola308 leaves exposed to salt stress. Taken together, these results suggest that the bacterial inoculation of canola increases salt tolerance by inducing an increase in the abundance of proteins related to glycolysis, tricarboxylic acid cycle, and amino acid metabolism.

Proteomic analysis of canola root inoculated with bacteria under salt stress.

Plant-growth promoting bacteria can ameliorate the negative effects of salt stress on canola. To better understand the role of bacteria in canola under salt stress, salt-sensitive (Sarigol) and salt-tolerant (Hyola308) cultivars were inoculated with Pseudomonas fluorescens and protein profiles of roots were compared. Bacterial inoculation increased the dry weight and length of canola roots under salt stress. Using a gel-free proteomic technique, 55 commonly changed proteins were identified in Sarigol and Hyola308 roots inoculated with bacteria under salt stress. In both canola cultivars, proteins related to amino acid metabolism and tricarboxylic acid cycle were affected. Hierarchical cluster analysis divided the identified proteins into three clusters. Proteins related to Clusters II and III, which were secretion-associated RAS super family 1, dynamin-like protein, and histone, were increased in roots of both Sarigol and Hyola308 inoculated with bacteria under salt stress. Based on pathway mapping, proteins related to amino acid metabolism and the tricarboxylic acid cycle significantly changed in canola cultivars inoculated with or without bacteria under salt stress. These results suggest that bacterial inoculation of canola roots increases tolerance to salt stress by proteins related to energy metabolism and cell division. BIOLOGICAL SIGNIFICANCE: Plant-growth promoting bacteria as an emerging aid can ameliorate the negative effect of salt stress on canola. To understand the role of bacteria in canola under salt stress, salt sensitive Sarigol and tolerant Hyola308 cultivars were used. Dry weight and length of canola root were improved by inoculation of bacteria under salt stress. Using gel-free proteomic technique, 55 commonly changed proteins identified in Sarigol and Hyola308 inoculated with bacteria under salt stress. In both canola cultivars, the number of proteins related to amino acid metabolism and tricarboxylic acid cycle was more than other categories with higher change in protein abundance. Hierarchical cluster analysis divided into 3 clusters. Cluster II including secretion-associated RAS super family 1 and dynamin-like protein and Cluster III including histones H2A were increased by bacterial inoculation in both cultivars. Furthermore, pathway mapping highlighted the importance of S-denosylmethionine synthetase and malate dehydrogenase that decreased in canola inoculated with bacteria under salt stress. These results suggest that bacterial inoculation helps the canola to endure salt stress by modulating the proteins related to energy metabolism and cell division.