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The effects of osmotic stress induced by polyethylene glycol on the seed germination of 16 walnut genotypes (‘Z30’, ‘Z53’, ‘Z67’, ‘Z60’, ‘Z63’, ‘K72’, ‘B21’, ‘V30’, ‘Panegine20’, ‘Hartley’, ‘Pedro’, ‘Vina’, ‘Lara’, ‘Serr’, ‘Ronde de Montignac’, and ‘Chandler’) of Juglans regia L. were studied. Potted seeds were kept under controlled conditions (12/12-h light/dark photoperiod and 25 ± 1 °C) during the experiments. The objective was to screen genotypes and determine the critical range of osmotic potential (ψS) for walnut seeds during germination. Decreasing the ψS of the germination solutions markedly reduced germination percentage in all genotypes, but there were variations in degree among the genotypes. The Z genotypes were the most sensitive to osmotic stress, and their germination rates were the lowest at ψS more negative than –0.75 MPa. Cluster analysis produced a dendrogram with four groups differing in their tolerance to osmotic stress. Based on factor analysis, four factors explained 90.45% of data total variance. Factor analysis showed that tissue fresh and dry weight, tissue water content, and thickness were the most important traits under drought condition. Regression analysis failed to show a significant relationship between percent germination and either seed weight (r2 = 0.0601) or kernel weight (r2 = 0.0258).

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There is a lack of information on the rhizosphere of nut-bearing trees where microbial populations can benefit roots and tree growth. The current research aimed at discovering plant growth-promoting rhizobacteria (PGPR) in the rhizosphere of soil samples from around the root zone of six walnut trees, each of which was considered as a genotype, i.e. ‘TT1’, ‘TT2’, ‘SS2’, ‘ZM1’, ‘Chandler’ and ‘Haward’. The trees grew in different arid and semiarid regions of Iran and Turkey. The strains were isolated and identified based on different morphological and biochemical markers. Drought-stress tolerance was assessed in the case of each isolate through their transfer to culture medium, containing polyethylene glycol (PEG6000) at 0 and 373.80 g L−1. Resilient strains were analyzed for measuring their ability to produce siderophore, hydrogen cyanide (HCN), Indole-3-acetic acid (IAA) and Gibberellic acid (GA3). In sum, 211 isolates were identified, of which a large number belonged to the Bacillus genus and, specifically, 78% of the strains were able to grow under drought stress conditions. The genus Arthrobacter was only detected in the rhizosphere of ‘ZM1’, ‘Haward’ and ‘TT1’ genotypes. In 4% of the strains, IAA production exceeded 53 mg L−1, while a high level of phosphorus solubility was verified in 6% of the strains. No strain was found to have the capability of producing HCN. The strains were screened for drought-tolerance, which resulted in the discovery of two promising strains, i.e. ZM39 and Cha43. Based on molecular identification through amplification and sequencing of the 16S rDNA gene, these two strains seemed to belong to Bacillus velezensis and Bacillus amyloliquefaciens, respectively. The discovery of new PGPR strains could probably assist walnut trees in improving their mechanisms of adaptation to drought stress.