Soil bacteria conferred a positive relationship and improved salt stress tolerance in transgenic pea (Pisum sativum L.) harboring Na+/H+ antiporter

Among grain legumes, peas (Pisum sativum L.) are highly sensitive to salt stress. Acclimatization of plants to such conditions is mandatory. We provide improved salt stress tolerance response of transgenic pea plants overexpressing the Na+/H+ gene from Arabidopsis thaliana and a positive association with salt-tolerant plant growth-promoting rhizobacteria (PGPR). In addition to salt stress tolerance and phosphate solubilization, the selected rhizobacterial isolates were identified for indole acetic acid and proline production ability. Seed germination percentage in transgenic pea plants was significantly higher under NaCl challenge. The wild-type (WT) pea plants inoculated with known numbers of viable cells of salt-tolerant PGPR and transgenic pea plants without any inoculation showed better growth performance under salt stress. However, the PGPR-inoculated transgenic plants showed significant increase in growth and biomass compared to the WT counterpart. An increase in antioxidant enzymes, i.e. Superoxide dismutase and peroxidases, was observed in PGPR-inoculated transgenic plants under salt stress. We could not see any negative effect of the transgene in pea plants on the growth of associated PGPR. The overall impact of microbe-mediated elicitation responses in transgenic plants, whether at the biochemical or molecular level, may lead to protection against salt stress.

Soil bacteria conferred a positive relationship and improved salt stress tolerance in transgenic pea (Pisum sativum L.) harboring Na+/H+ antiporter

Among grain legumes, peas (Pisum sativum L.) are highly sensitive to salt stress. Acclimatization of plants to such conditions is mandatory. We provide improved salt stress tolerance response of transgenic pea plants overexpressing the Na+/H+ gene from Arabidopsis thaliana and a positive association with salt-tolerant plant growth-promoting rhizobacteria (PGPR). In addition to salt stress tolerance and phosphate solubilization, the selected rhizobacterial isolates were identified for indole acetic acid and proline production ability. Seed germination percentage in transgenic pea plants was significantly higher under NaCl challenge. The wild-type (WT) pea plants inoculated with known numbers of viable cells of salt-tolerant PGPR and transgenic pea plants without any inoculation showed better growth performance under salt stress. However, the PGPR-inoculated transgenic plants showed significant increase in growth and biomass compared to the WT counterpart. An increase in antioxidant enzymes, i.e. Superoxide dismutase and peroxidases, was observed in PGPR-inoculated transgenic plants under salt stress. We could not see any negative effect of the transgene in pea plants on the growth of associated PGPR. The overall impact of microbe-mediated elicitation responses in transgenic plants, whether at the biochemical or molecular level, may lead to protection against salt stress.

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  • Ali Z, Shumacher HM, Dobbernack EH, El-Banna A, Hafeez FY, Jacobsen HJ, Kiesecker H (2010). Dicistronic binary vector system - a versatile tool for gene expression studies in cell cultures and plants. J Biotechnol 145: 9–16.
  • Alscher RG, Donahue JC, Cramer CL (1997). Reactive oxygen species and antioxidants: relationship in green cells. Plant Physiol 100: 224–233.
  • Apse MP, Aharon GS, Snedden WA, Blumwald E (1999). Salt tolerance conferred by over expression of a vacuolar Na+/H+ antiport in Arabidopsis. Science 285: 1256–1258.
  • Ashraf M, Foolad MR (2007). Improving plant abiotic-stress resistance by exogenous application of osmo-protectants glycine betaine and proline. Environ Exp Bot 59: 206–216.
  • Auge RM (2001). Water relations, drought and vesicular–arbuscular mycorrhizal symbiosis. Mycorrhiza 11: 3–42.
  • Bates L, Waldern SR, Teare ID (1973). Rapid determination of free proline for water stress studies. Plant Soil 39: 205–207.
  • Beauchamp C, Fridovich I (1971). Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Anal Biochem 44: 276–287.
  • Bianco C, Defez R (2009). Medicago truncatula improves salt tolerance when nodulated by an indole-3-acetic acid overproducing Sinorhizobium meliloti strain. J Exp Bot 60: 3097–3107.
  • Bor M, Özdemir F, Türkan I (2003). The effect of salt stress on lipid peroxidation and antioxidants in leaves of sugar beet Beta vulgaris L. and wild beet Beta maritima L. Plant Sci 164: 77–84.
  • Chen T, Yang Q, Zhang X, Ding W, Gruber M (2012). An alfalfa (Medicago sativa L.) ethylene response factor gene, MsERF11, enhances salt tolerance in transgenic Arabidopsis. Plant Cell Rep 31: 1737–1746.
  • Creus CM, Sueldo RJ, Barassi CA (2004). Water relations and yield in Azospirillum-inoculated wheat exposed to drought in the field. Can J Botany 82: 273–281.
  • de Zelicourt A, Al-Yousif M, Hirta H (2013). Rhizosphere microbes as essential partners for plant stress tolerance. Mol Plant 6: 242–245.
  • Dimkpa C, Weinand T, Asch F (2009). Plant–rhizobacteria interactions alleviate abiotic stress conditions. Plant Cell Environ 32: 1682–1694.
  • Dodd IC, Perez-Alfocea F (2012). Microbial amelioration of crop salinity stress. J Exp Bot 63: 3415–3428.
  • Edi-Premono M, Moawad AM, Vleck PLG (1996). Effect of phosphate solubilizing Pseudomonas putida on the growth of maize and its survival in rhizosphere. Indones J Crop Sci 11: 13–23.
  • El-Banna A, Hajirezaei MR, Wissing J, Ali Z, Vaas L, Heine- Dobbernack E, Jacobsen HJ, Schumacher HM, Kiesecker H (2010). Over-expression of PR-10a leads to increased salt and osmotic tolerance in potato cell cultures.  J Biotechnol 150: 277–287.
  • Francois LE, Maas EV (1994). Crop response soils to management on salt-affected. In: Pessarakli M, editor. Hand Book of Plant and Crop Stress. New York, NY, USA: Marcel Dekker, pp. 149–181.
  • Gao S, Ouyang C, Wang S, Xu T, Tang L, Chen F (2008). Effects of salt stress on growth, antioxidant enzyme and phenylalanine ammonialyase activities in Jatropha curcas L. seedlings. Plant Soil Environ 54: 374–381.
  • Glick BR (1995). The enhancement of plant growth by free-living bacteria. Can J Microbiol 41: 109–117.
  • Glickman E, Dessaux Y (1995). A critical examination of the specificity of the Salkowski reagent for indolic compounds produced by phytopathogenic bacteria. Appl Environ Microbiol 61: 793–796.
  • Golpayegani A, Tilebeni HG (2011). Effect of biological fertilizers on biochemical and physiological parameters of basil (Ocimum basilicum L.) medicine plant. American-Eurasian J Agric Environ Sci 11: 411–416.
  • Grieve AM, Prior DL, Bevington BK (2007). Long-term effect of saline irrigation water on growth, yield, and fruit quality of Valencia orange trees. Plant Soil 45: 313–317.
  • Gururani M, Upadhyaya C, Baskar V, Venkatesh J, Nookaraju A, Park S (2013). Plant growth promoting rhizobacteria enhance abiotic stress tolerance in Solanum tuberosum through inducing changes in the expression of ROS scavenging enzymes and improved photosynthetic performance. J Plant Growth Regul 32: 245–258.
  • Gutierrez CK, Matsui GY, Lincoln DE, Lovell CR (2009). Production of the phytohormone indole-3-acetic acid by estuarine species of the genus Vibrio. Appl Environ Microbiol 75: 2253–2258.
  • Han HS, Lee KD (2005). Plant growth-promoting rhizobacteria: effect on antioxidant status, photosynthesis, mineral uptake and growth of lettuce under soil salinity. Res J Agric Biol Sci 1: 210–215.
  • Herzog V, Fahimi H (1973). A new sensitive colorimetric assay for peroxidase using 3,3’-diaminobenzidine as hydrogen donor. Anal Biochem 55: 554–562.
  • Jenkins GM, Rambaut A, Pybus OG, Holmes EC (2002). Rates of molecular evolution in RNA viruses: a quantitative phylogenetic analysis. J Mol Evol 54: 156–165.
  • Kim YC, Leveau J, Gardener BBM, Pierson EA, Pierson LS 3rd, Ryu CM (2011). The multifactorial basis for plant health promotion by plant-associated bacteria. Appl Environ Microbiol 77: 1548–1555.
  • Kloepper JW (1992). Plant growth promoting rhizobacteria as biological control agents. In: Metting Jr FB, editor. Soil Microbial Ecology: Applications in Agricultural and Environmental Management. New York, NY, USA: Marcel Dekker, pp. 255–274.
  • Kumari S,  Vaishnav A,  Jain S, Varma A,  Choudhary DK (2015). Bacterial-mediated induction of systemic tolerance to salinity with expression of stress alleviating enzymes in soybean (Glycine max L. Merrill). J Plant Growth Regul 34: 558–573.
  • Lane DJ (1991). 16S/23S rRNA sequencing. In:  Stackebrandt E, Goodfellow M, editors. Nucleic Acid Techniques in Bacterial Systematics.  New York, NY, USA: John Wiley and Sons, pp. 115–175.
  • Mandhania S, Madan S, Sawhney V (2006). Antioxidant defense mechanism under salt stress in wheat seedlings. Biol Plantarum 50: 227–231.
  • Mayak S, Tirosh T, Glick BR (2004). Plant growth-promoting bacteria confer resistance in tomato plants to salt stress. Plant Physiol Biochem 42: 565–572.
  • Miller JH (1972). Experiments in Molecular Genetics. Cold Spring Harbor, NY, USA: Cold Spring Harbor Laboratory Press.
  • Mittova V, Guy M, Tal M, Volokita M (2002). Response of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii to salt-dependent oxidative stress: increased activities antioxidant enzymes in roots plastids. Free Radic Res 36: 195– 202.
  • Montesinos E, Bonaterra A, Badosa E, Francés J, Alemany J, Llorente I, Moragrega C (2002). Plant-microbe interactions and the new biotechnological methods of plant disease control. Int Microbiol 5: 169–175.
  • Munns R, Tester M (2008). Mechanisms of salinity tolerance. Ann Rev Plant Bio 59: 651–681.
  • Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497.
  • Naz I, Bano A (2010). Biochemical, molecular characterization and growth promoting effects of phosphate solubilizing Pseudomonas sp. isolated from weeds grown in salt range of Pakistan. Plant Soil 334: 199–207.
  • Noreen Z, Ashraf M, Hassan MU (2007). Inter-accessional variation for salt tolerance in pea (Pisum sativum L.) at germination and seedling stage. Pak J Bot 39: 275–285.
  • Patten CL, Glick BR (2002). Role of Pseudomonas putida indole acetic acid in the development of the host plant root system. Appl Environ Microbiol 68: 3795–3801.
  • Pikovskaya RI (1948). Mobilization of phosphorus in soil in connection with the vital activity of some microbial species. Mikrobiologiya 17: 362–370.
  • Richter A, Jacobsen HJ, de Kathen A, de Lorenzo G, Briviba K, Hain R, Ramsay G, Kiesecker H (2006). /Transgenic peas (Pisum sativum) expressing polygalacturonase inhibiting protein from raspberry (Rubus idaeus) and stilbene synthase from grape (Vitis vinifera). Plant Cell Rep 25: 1166–1173.
  • Sambrook J, Fritsch EF, Maniatis T (1989). Molecular Cloning: A Laboratory Manual. 3rd ed. Cold Spring Harbor, NY, USA: Cold Spring Harbor Laboratory Press.
  • Shahid MA, Pervez MA, Balal RM, Mattson NS, Rashid A, Ahmad R, Ayyub CM, Abbas T (2011). Brassinosteroid (24-epibrassinolide) enhances growth and alleviates the deleterious effects induced by salt stress in pea (Pisum sativum L.). Aust J Crop Sci 5: 500–510.
  • Shukla PS, Agarwal PK, Jha B (2012). Improved salinity tolerance of Arachis hypogaea (L.) by the interaction of halotolerant plant growth promoting rhizobacteria. J Plant Growth Regul 31: 195–206.
  • Somers DA, Samac DA, Olhoft PM (2003). Recent advances in legume transformation. Plant Physiol 131: 892–899.
  • Somers E, Vanderleyden J, Srinivasan M (2004). Rhizosphere bacterial signaling: a love parade beneath our feet. Crit Rev Microbiol 30: 205–240.
  • Steel KJ (1961). The oxidase reaction as a toxic tool. J Gen Microbiol 25: 297–306.
  • Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 30: 2725–2729.
  • Turner S, Pryer KM, Miao VPW, Palmer JD (1999). Investigating deep phylogenetic relationships among cyanobacteria and plastids by small subunit rRNA sequence analysis. J Eukaryot Microbiol 46: 327–338.
  • Upadhyay SK, Singh JS, Saxena AK, Singh DP (2012). Impact of PGPR inoculation on growth and antioxidant status of wheat under saline conditions. Plant Biol 14: 605–611.
  • Vadez V, Rashmi M, Sindhu K, Muralidharan M, Pushpavalli R, Turner NC, Krishnamurthy L, Gaur PM, Colmer TD (2012). Large number of flowers and tertiary branches, and higher reproductive success increase yields under salt stress in chickpea. Eur J Agron 41: 42–51.
  • Zahir AZ, Ghani U, Naveed M, Nadeem SM, Asghar HN (2009). Comparative effectiveness of Pseudomonas and Serratia sp. containing ACC-deaminase for improving growth and yield of wheat (Triticum aestivum L.) under salt-stressed conditions. Arch Microbiol 191: 415–424.
  • Zhang HX, Blumwald E (2001). Transgenic salt-tolerant tomato plants accumulate salt in foliage but not in fruit. Nat Biotechnol 19: 765–768. 1 2
  • Supplementary Figure 3. Effect of NaCl stress (100 mM) and PGPR
  • interaction on root percentage of water contents of transgenic and
  • wild type pea plants. The experiment was performed in vitro in
  • tissue culture jars containing half-strength MS media in a culture
  • room at 25 ± 2 °C with a 16/8-h light/dark cycle. Bac-A and Bac-B
  • stand for isolate PS2 and PS5, respectively. The data were analyzed
  • by two-way ANOVA for treatments and varieties. Data are presented
  • for each sample corresponding to three independent replicates, and
  • each replicate was the pool of the three plantlets of the jars. Bars
  • show standard errors (mean ± SE). All means sharing a common
  • letter are similar; otherwise, means differ significantly at P < 0.05.
Turkish Journal of Botany-Cover
  • ISSN: 1300-008X
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
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