Soil Biology & Biochemistry 35 (2003) 973978 www.elsevier.

com/locate/soilbio

Inuence of growth-promoting bacteria on the growth of wheat in different soils and temperatures
Dilfuza Egamberdiyevaa,*, Gisela Hoichb
b a Institute of Microbiology, Uzbek Academy of Sciences, Abdulla Qadiriy 7 B, Tashkent 700128, Uzbekistan Centre for Agricultural Landscape and Land Use Research, Institute for Primary Production and Microbial Ecology, Eberswalder Str. 84, D-15374 Muncheberg, Germany

Received 16 August 2001; received in revised form 26 February 2003; accepted 11 March 2003

Abstract Plant-growth-promoting bacteria isolated from the rhizosphere, phyllosphere and soil of the root zone in different climatic regions of Germany and Uzbekistan were analysed for plant-growth-promoting effects and nutrient uptake on winter wheat on different soils and under different temperature regimes. The investigations were carried out in pot experiments using loamy sand and sandy loam soils from Muncheberg, Germany and Calcisol soil from Tashkent, Uzbekistan. The temperature and soil types were found to inuence growthpromoting effects. Inoculation with bacterial strains Pseudomonas uorescens PsIA12, Pantoea agglomerans 050309 and Mycobacterium sp. 44 isolated from Muncheberg (semi-continental climate) was found to signicantly increase the root and shoot growth of winter wheat at 16 8C compared to 26 8C in loamy sand. Mycobacterium phlei MbP18 and Mycoplana bullata MpB46 isolated from Tashkent (semi-arid climate) were found to signicantly increase the root and shoot growth of winter wheat in nutrient-poor Calcisol at 38 8C as well as in nutrient-rich loamy sand at 16 8C. Bacterial inoculation also resulted in signicantly higher N, P, and K contents of plant components. The bacteria isolates were able to survive in the rhizosphere and in the soil of winter wheat after root and shoot inoculation. q 2003 Elsevier Science Ltd. All rights reserved.
Keywords: Plant growth promoting bacteria; Rhizosphere; Soil type; Temperature

1. Introduction Soil bacteria that colonize plant roots and promote growth when added to seeds, roots or tubers have been termed plant-growth-promoting rhizobacteria (PGPR) (Kloepper et al., 1980). Different plant-growth promoting rhizosphere bacteria, including associative bacteria such as Azospirillum, Bacillus, Pseudomonas, Enterobacter groups have been used for their benecial effects on plant growth (Kloepper and Beauchamp, 1992; Hoich et al., 1994). The mechanisms of plant growth stimulation by associative bacteria are mobilization of nutrients (Lifshitz et al., 1987), stimulation of root growth by production of phytohormones (Bothe et al., 1992; Kloepper et al., 1980) and antagonism against soil borne plant pathogens (Kloepper et al., 1988a; Hoich et al., 1994). All of these mechanisms presuppose a direct contact between bacteria and the root surface and
* Corresponding author. Fax: 998-712-417129. E-mail address: dilfuza_egamberdiyeva@yahoo.com Egamberdiyeva).

(D.

an active status of the introduced bacteria. The survival of inoculated PGPR in the plant rhizosphere is in most cases a precondition for a potential plant stimulation effect during the vegetation time or at least during early plant develop ment (Hoich et al., 1995). Several studies clearly showed the effect of plant growth promoting bacteria on plant growth of different crops at different climates, soils and temperatures. (Bilal et al., 1993; Hoich and Kuhn, 1996; Javed and Arshad, 1997; Bashan and Holguin, 1997). Knowledge of the plant growth promoting effects and the survival of the plant-growth-promoting bacteria at different temperatures and soils may be important for successful root inoculation. However, research to compare the effects of key ecological factors, i.e. temperature and soil types, on plant growth promoting efciency of selected bacteria for winter wheat in different climatic regions has not been performed. The major objectives of our research were to compare the effect of selected plant-growth-promoting bacteria isolated from the rhizosphere, phyllosphere and soil of the root zone of different crops on the growth of

0038-0717/03/$ - see front matter q 2003 Elsevier Science Ltd. All rights reserved. doi:10.1016/S0038-0717(03)00158-5

974

D. Egamberdiyeva, G. Hoich / Soil Biology & Biochemistry 35 (2003) 973978

winter wheat in Germany and Uzbekistan at different temperatures and soils.

2. Materials and methods 2.1. Plant and soil The experiments were carried out using loamy sand from Muncheberg and a sandy loam from Dedelow, Germany, and a Calcisol, from Tashkent, Uzbekistan. The soil chemical and physical properties are presented in Table 1. Winter wheat (Triticum aestivum) cv. Bussard, (Germany), cv. Ziklon, (Uzbekistan) were used as the tests plants for the inoculation experiments. Seeds of wheat were obtained from the Centre for Agricultural Landscape and Land Use Research, Muncheberg, Germany and from the University of Agriculture of Uzbekistan, Tashkent. 2.2. Microorganisms The bacterial strains used were isolated from the following plants: Pseudomonas agglomerans 030509 from the phyllosphere of triticale, Mycobacterium sp. 44 from the phyllosphere of pea grown in loamy sand (Muncheberg, eld) and Mycoplana bullata MpB46 from soil of the root zone of maize, Mycobacterium phlei MbP18 from soil of the root zone of winter wheat grown in Calcisol soil (Tashkent, eld). P. uorescens PsIA 12 were isolated from the rhizosphere of wheat by Hoich et al. (1995) in previous works and used for this experiments in comparison with new isolated strains. For isolation of bacteria from the rhizosphere 1 g of washed roots and bacteria from the phyllosphere 1 g leaves were macerated and shaken with 10 ml sterile water. For isolation of bacteria from the soil of the root zone, 1 g soil from the root surface were shaken with 10 ml sterile water. The resulting suspensions were spread over the surface of a glycerol peptone agar: Peptone 10 g, glycerol 10 ml, NaCl 5 g, KH2PO4 0,1 g, agar 15 g/l sterile water. After an incubation time of four days at 28 8C the bacterial strains were isolated from the plates and identied. The identication scheme based on a combination of conventional test like morphological, physiological and chemotaxonomic

characterization (Holt et al., 1994) and the use of commercial identication systems like Biolog (GN, GP) from the Biolog Inc. and API 20E or API 20NE (bioMerieux), (Behrendt et al., 1997). Mutants of Mycobacterium sp. 44, and M. bullata MpB46 marked with antibiotic resistance were obtained after plating of the parental strain onto glycerol peptone agar amended with rifampicin (120 mg l21). After an incubation time of four days at 28 8C the rifampicin resistant strains were selected based on similarities in colony morphology and growth rate with the parent strains Mycobacterium sp. 44, and M. bullata MpB46, and were recultured on medium containing rifampicin to ensure stability of the antibiotic resistance marker. 2.3. Plant growth and inoculation in pots The study of the effect of bacterial strains isolated from Germany on plant growth and nutrient uptake of wheat was carried out in a temperature regulated growth chamber using a nutrient-rich loamy sand and a sandy loam. Plastic containers (5 cm diameter and 18 cm deep) with 350 g soil were incubated at a light intensity 20kLux for 16 h at 16 8C during the day and 12 8C at night, and at 24kLux with 26 8C at day and at 16 8C night (Germany). The experiments with bacterial strains isolated from Uzbekistan were carried out in pot experiments using loamy sand and nutrient poor Calcisol soil in the greenhouse. Plants were grown in pots (10 cm diameter and 12 cm deep) lled with 500 g soil for four weeks under open natural conditions with a temperature of 36 38 8C during the day and 20 24 8C at night in summer (July, Uzbekistan), and under greenhouse conditions 17 20 8C during the day and 8 12 8C at night in spring (March, Germany). The inoculation treatments were set-up in a randomized design with six replicates. Five seeds of wheat were sown per pot and after germination, plants were thinned to four per pot. The bacteria were grown in glycerol peptone-medium. Tubes were secured on a rotary shaker (120 rpm; 23 8C) and agitated for three days. Seedlings of these plants were inoculated with 1 ml of the bacterial suspension that resulted in an inoculum density of 106 107 cfu ml21. Control seeds received 1 ml glycerol peptone-medium.

Table 1 Soil chemical properties, and soil particle distribution of the top soil layer (030 cm) Site Type Ctot (mg 100 g21) Ntot (mg 100 g21) Ptot (mg 100 g21) K (mg 100 g21) Mg (mg 100 g21) pH Soil particle size (mm) 2.00.2% Muencheberg Dedelow Tashkent Loamy sand Sandy loam Calcisol 700 800 200 60 80 6 6.2 7.1 3.0 7.4 8.4 12.0 3.7 6.5 6.0 6.9 6.1 8.5 7.6 4.3 2.2 0.20.02% 79.8 70.2 54.5 ,0.02% 12.6 25.5 43.3

 D. Egamberdiyeva, G. Hoich / Soil Biology & Biochemistry 35 (2003) 973978

975

The soil was moistened with water and maintained at 60% of moisture holding capacity (MHC). Four weeks after germination, shoots and roots were separated and dried overnight at 105 8C before determining the root and shoot dry weight. Root and shoot dry matter were assayed in six-leaves-stage and N, P, K content of plants as criteria for growth promotion. 2.4. Survival of antibiotic resistant bacteria after inoculation The investigations were conducted with antibioticresistant mutants of the Mycobacterium sp. 44 and M. bullata MpB46. Plant growth and inoculation experiments were used same as described above. Plants were harvested 28 days after the emergence of the seeds. After harvesting, adhering soil was removed from wheat roots and shoots were separated. For determination of the soil colonization, 1 g soil removed from the roots was shaken with 10 ml sterile water and 40% tetramethylthiuramdisulphide (TMTD) solution for 30 min. For determination of rhizosphere colonization 1 g of washed roots were macerated and shaken with 10 ml sterile water. To determine the phyllosphere colonization 1 g leaves were macerated and shaken with 10 ml sterile water. The resulting suspensions were evaluated for colony forming units (cfu) according to the dilution-plate method in glycerol peptone agar with addition of 120 ppm rifampicin. By addition of TMTD and rifampicin, the native fungal and bacterial ora was mostly excluded from the plates. After incubation of seven days at 28 8C, the reisolated, rifampicin resistant strains were identied for colony characteristics and compared against the parent strains. 2.5. Statistical analysis The data were analysed for signicant differences p , 0:05 of main effects using a two-way ANOVA and StudentNewman-Keuls.

Table 2 The inuence of Pseudomonas uorescens PsIA12 and Pantoea agglomerans 050309 on root and shoot dry matter of winter wheat in different soils (incubation: 16 8C, control 100%; loamy sand: 126,6 mg shoot, 73,4 mg root; sandy loam: 126,3 mg shoot, 86,4 mg root) Bacterial strains Loamy sand Shoot Root Sandy loam Shoot Root

Control 100(126,6)a 100(73,4)a 100(126,3)a 100(86,4)a Pseudomonas 123* 130* 119* 111 uorescens PsIA12 Pantoea 118* 114* 105 100 agglomerans 050309 LSD a 0.05 13 20 12 25 * Signicantly different from the control for P , 0:05. a mg/pot.

was no effect on the N contents of shoots (Fig. 1). Inoculation experiments with winter wheat at different temperatures showed that plant-growth promoting bacteria Mycobacterium sp. 44 (originating from semi-continental climate) was more effective at 16 8C as compared to incubation at 26 8C (Fig. 2).

Fig. 1. Inoculation effect of Pseudomonas uorescens PsIA12 and Pantoea agglomerans 050309 on P and K uptake of winter wheat in loamy sand (control 100%; incubation: 16 8C; nutrient contents: 0,22 mg P/shoot, 0,29 mg P/root; 2,5 mg K/shoot, 1,2 mg K/root).

3. Results 3.1. Growth promotion of winter wheat by bacterial inoculants from Germany at different soils and temperatures Bacterial inoculation affected the early plant growth and the nutrient content of winter grown in different soils and temperatures. The inoculation of winter wheat grown in the loamy sand with bacteria strains P. uorescens PsIA12 and P. agglomerans 050309 (originating from semi-continental climate) signicantly p , 0:05 increased the shoot and root dry weights from 18 to 30% as compared to the control. This growth promoting effect was higher in the loamy sand than in the sandy loam (Table 2). P. uorescens PsIA12 and P. agglomerans 050309 signicantly increased the P- and K-uptake of winter wheat grown at the loamy sand, but there

Fig. 2. The inuence of Mycobacterium sp. 44 on growth of winter wheat at different temperatures in loamy sand (control 100%; incubation 16 8C: 111,9 mg shoot, 217,0 mg root; incubation 26 8C: 126,6 mg shoot, 73,4 mg root).

976

D. Egamberdiyeva, G. Hoich / Soil Biology & Biochemistry 35 (2003) 973978

26 8C. However, the survival of inoculated bacteria strains originating from semi-continental climate was much better at 16 8C than at 26 8C. The strain M. bullata MpB46 isolated from semi-arid climate, colonized the rhizosphere and the soil of winter wheat at 26 8C better than at 16 8C (Table 3). Moreover, the strains were unable to survive in the phyllosphere, whereas M. bullata MpB46 showed very low colonization efciency in the phyllosphere.

4. Discussion
Fig. 3. Inoculation effect of Mycoplana bullata MpB 46 and Mycobacterium phlei MbP18 on shoot, root dry matter and nutrient uptake of winter wheat in Calcisol (pot experiment, incubation: 36 8C, control 100%, (DM-dry matter), 392,7 mg shoot, 268,1 mg root; nutrient contents: 13,1 mg N/shoot, 4,2 mg N/root; 1,8 mg P/shoot, 0,9 mg P/root; 21,4 mg K/shoot, 5,8 mg K/root).

3.2. Growth promotion of winter wheat by bacterial inoculants from Uzbekistan at different soils and temperatures The bacterial strain Mycobacterium phlei MbP18 and M. bullata MpB46 (originating from semi-arid climate, Uzbekistan) was effective for winter wheat in the nutrient poor Calcisol (Fig. 3). There were no effects on plant growth and nutrient uptake of wheat in nutrient-rich loamy sand at 16 8C (data not shown). The strain M. bullata MpB46 increased root and shoot dry matter of winter wheat (12%) signicantly as compared to the control (Fig. 3). The inoculation of seedlings with M. phlei MbP18 signicantly increased the shoot, root dry weight and contents (N, P, K) of winter wheat in Calcisol soil from 13 to 53%. The bacteria strains were not effective in loamy sand (data not shown). 3.3. Survival and establishment of antibiotic resistant bacteria in winter wheat at different temperatures Rifampicin-resistant isolates derived from Mycobacterium sp. 44 and M. bullata MpB46 were tested for their ability to colonize wheat roots, shoots and in soil at different temperatures. The two inoculated bacteria Mycobacterium sp. 44 and M. bullata MpB 46 were able to establish a population in the rhizosphere and in the soil of winter wheat up to four weeks after sowing at temperatures of 16 and

The experiments have demonstrated that independent of the origin, selected growth stimulating bacteria isolates (rhizosphere, phyllosphere, soil of the root zone) are able to increase the growth and nutrient uptake of winter wheat in loamy sand, sandy loam and Calcisol soils at different temperatures. The importance of physiological plant promotion characteristics may vary with soil and climatic parameters (Hoich et al., 1994) and may partly be affected by different growth stimulation effects following inoculation. The bacteria isolates P. uorescens PsIA12 and P. agglomerans 050309 signicantly increased the plant growth of winter wheat in loamy sand compared to the sandy loam. Hoich et al. (1995, 1997) also demonstrated in her previous work, that strains P. uorescens PsIA12 and Agrobacterium rhizogenes A1A4 promoted the growth of legume, maize and wheat under eld conditions on loamy sand much better than sandy loam. Our bacteria isolates M. phlei MbP18 and M. bullata MpB46 from semi-arid climate had a much better effect on winter wheat in nutrient-poor Calcisol soil than in nutrient-rich loamy sand. Defreitas and Germida (1992a) also demonstrated that in low fertility Asquith soil, pseudomonas bacteria strains signicantly enhanced early plant growth. According to Paula et al. (1992), the magnitude of the plant response to any microbial inoculation can be greatly affected by the nutrient content of soil. Inoculation of plants with bacteria only marginally increased yields when tested under ideal climatic situations. The greatest benets occurred when crops encountered stressful conditions for prolonged periods (Lazarovits and Norwak, 1997), e.g. when lodgepole pine seedlings were treated with PGPR beneted from bacterisation only at the poorest site used for transplanting (Chanway and Holl, 1994). Non-treated plants by comparison performed poorly under such conditions. In our study, statistically signicant

Table 3 Colonization of M. bullata MpB46 and Mycobacterium sp. 44 in the soil, rhizo- and phyllosphere of winter wheat after inoculation (loamy sand, values in 103 cfu 1 g21 of fresh root, shoot and soil) Bacterial strains Temperature (16 8C) Soil of the root zone Mycobacterium sp. 44 M. bullata MpB46 7.2 17.0 Rhizo-sphere 498.0 83.4 Phyllo-sphere 1.5 0 Temperature (26 8C) Soil of the root zone 5.8 1.8 Rhizo-sphere 220.0 444.0 Phyllo-sphere 1.4 0

 D. Egamberdiyeva, G. Hoich / Soil Biology & Biochemistry 35 (2003) 973978

977

enhancement of wheat growth was observed at moderate temperature 16 8C, rather than 26 8C. According to Hoich et al. (1994) and Hoich and Kuhn (1996), Pseudomonas sp., Rhizobium sp. and Agrobacterium sp. isolated from sites at semi-continental climate promoted the growth of young plants and increased the yields of gramineae and legumes under the respective eld conditions. One strain M. phlei MbP18, isolated from a semi-arid climate increased signicantly plant growth and nutrient uptake of winter wheat at 38 8C compared to growth at 16 8C. Javed and Arshad (1997) also isolated bacteria, which stimulated the plant growth of wheat and rice in warm climates. Paau (1989) described that for obtaining competitive and effective bacterial strains, these must be screened and isolated from the pool of indigenous soil bacteria, which supposedly are adapted to the particular climatic conditions of the site. Waldon et al. (1989) also demonstrated that rhizobial bacteria isolated from nodules of the desert woody legumes Prosopis glandulosa grew better at 36 8C than at 26 8C. They are physiologically distinct, suggesting that these bacteria are adaptive to their respective environmental conditions. Bacterial inoculation also signicantly increased nutrient contents by plants. Increased nutrient uptake by plants inoculated with plant-growth promoting bacteria has been attributed to the production of plant growth regulators at the root interface, which stimulated root development and resulted in better absorption of water and nutrients from the soil (Kloepper et al., 1991; Zimmer et al., 1995; Hoich and Kuhn, 1996). In our experiments, auxin was detected in all bacterial suspensions, expect P. agglomerans 050309. As successful plant growth promoting inoculants, bacteria must be able to rapidly colonize the root system during the growing season (Defreitas and Germida, 1992b). Our results showed that Mycobacterium sp. 44, and M. bullata MpB46 were able to colonize the rhizosphere and the soil of winter wheat. Defreitas and Germida (1992b) also demonstrated similar effects for wheat using growth promoting Pseudomonas strains. The survival of the strains Mycobacterium sp. 44 (from semi-continental climate) was much better at 16 8C than 26 8C, whereas the bacterial strain M. bullata MpB46 (from semi-arid climate) was more effective at 26 8C than 16 8C. These results demonstrate that the ambient temperature inuences the root colonization activity of introduced strains. From the results we conclude that effective plantgrowth promoting bacteriaplant systems must be tested and established in controlled vegetation experimental designs with consideration of the specic ecological site conditions of practical applications (soil type, temperature). However, the extent of stimulation of plants by tested bacterial strains from Uzbekistan and the persistence of plant growth promoting activity under actual eld conditions remains unclear. Thus, the experiments concerning

stimulation of winter wheat must be followed by investigations under eld conditions.

Acknowledgements We thank Monika Roth, Irina Ba r for technical assistance. This work was nanced by German Academic Exchange Service (DAAD), the Center of Agricultural Landscape and Land Use Research (ZALF) Muencheberg, and the Humboldt University of Berlin, Institute of Plant Production.

References
Bashan, Y., Holguin, G., 1997. Azospirillum-plant relationships: environmental and physiological advances (1990 1996). Canadian Journal of Microbiology 43, 103 121. Behrendt, U., Muller, Th., Seyfarth, W., 1997. The inuence of extensication in grassland management on the populations of microorganisms in the phyllosphere of grasses. Microbiological Research 152, 75 85. Bilal, R., Rasul, G., Malik, K.A., 1993. Attachment, colonization and proliferation of Azospirillum brasiliense and Enterobacter spp. on root surface of grasses. World Journal of Microbiology and Biotechnology 9, 63 69. Bothe, H., Korsgen, H., Lehmacher, T., Hundeshagen, B., 1992. Differential effects of Azospirillum, auxin and combined nitrogen on growth of the roots of wheat. Symbiosis 13, 167 179. Chanway, C.P., Holl, F.B., 1994. Ecological growth response specicity of two Douglas-r ecotypes inoculated with coexistent benecial bacteria. Canadian Journal of Botany 72, 582586. Defreitas, J.R., Germida, J.J., 1992a. Growth promotion of winter wheat uorescent Pseudomonas under growth chamber conditions. Soil Biology and Biochemistry 24, 11271135. Defreitas, J.R., Germida, J.J., 1992b. Growth promotion of winter wheat uorescent Pseudomonas under eld conditions. Soil Biology and Biochemistry 24, 11371146. Hoich, G., Kuhn, G., 1996. Forderung das Wachstums und der Nahrstoffaufnahme bei kurziferen Ol- und Zwischenfruhten durch inokulierte Rhizospherenmikroorganismen. Zeischrift fur Panzener nahrung und Bodenkunde 159, 575 578. Hoich, G., Wiehe, W., Kuhn, G., 1994. Plant growth stimulation with symbiotic and associative rhizosphere microorganisms. Experientia 50, 897 905. Hoich, G., Wiehe, W., Heght-Buchholz, C.H., 1995. Rhizosphere colonization of different growth promoting Pseudomonas and Rhizobium bacteria. Microbiological Research 150, 139147. Hoich, G., Tappe, E., Kuhn, G., Wiehe, W., 1997. Einu associativer Rhizospharenbakterien auf die Nahrstoffaufnahme und den Ertrag von Mais. Archive Acker und Panzen Boden 41, 323333. Holt, J.G., Krieg, N.R., Sneath, P.H., Staley, J.T., Williams, S.T., 1994. Bergeys Manual of Determinative Bacteriology, 9th ed, Williams and Wilkins, Baltimore. Javed, M., Arshad, M., 1997. Growth promotion of two wheat cultivars by plant growth promoting rhizobacteria. Pakistan Journal of Botany 29, 243 248. Kloepper, J.W., Beauchamp, C.J., 1992. A review of issues related to measuring of plant roots by bacteria. Canadian Journal of Microbiology 38, 12191232. Kloepper, J.W., Leong, J., Teintze, M., Schroth, M.N., 1980. Enhanced plant growth by siderophore produced by plant growth-promoting rhizobacteria. Nature (London) 286, 885886.

978

D. Egamberdiyeva, G. Hoich / Soil Biology & Biochemistry 35 (2003) 973978 Paau, M.A., 1989. Improvement of Rhizobium inoculants. Applied and Environmental Microbiology 55, 862 865. Paula, M.A., Urquiaga, S., Siqueira, I.O., Dobereiner, J., 1992. Synergistic effects of vesicular-arbuscular mycorrhizal fungi and diazotroc bacteria on nutrition and growth of sweet potato (Ipomoea batatas). Biology and Fertility of Soils 14, 6166. Waldon, H.B., Jenkins, M.B., Virginia, R.A., Harding, E.E., 1989. Charcteristics of woodland rhizobial population from surface- and deep-soil environments of the Sonoran Desert. Applied and Environmental Microbiology 55, 30583064. Zimmer, W., Kloos, K., Hundeshagen, B., Neiderau, E., Bothe, H., 1995. Auxin biosynthesis and denitrication in plant growth promotion bacteria. In: Fendrik, J., De Gallo Vandeleyden, J., De Zamoroczy, D. (Eds.), Azospirillum VI and related microorganisms, Series G: Ecological, 37., pp. 120 141.

Kloepper, J.W., Hume, D.J., Schner, F.M., Singleton, C., Tipping, B., Laliberte, M., Frauley, K., Kutchaw, T., Simonson, C., Lifshitz, R., Zaleska, I., Lee, L., 1988a. Plant growth promoting rhizobacteria on canola. Plant Diseases 72, 4246. Kloepper, J.W., Zablowicz, R.M., Tipping, B., Lifshitz, R., 1991. Plant growth mediated by bacterial rhizosphere colonizers. In: Keister, D.L., Gregan, B. (Eds.), The rhizosphere and plant growth, 14. BARC Symposium, pp. 315326. Lazarovits, G., Norwak, J., 1997. Rhizobacteria for improvement of plant growth and establishment. Horticultural Science 32, 188192. Lifshitz, R., Kloepper, J.W., Kozlowski, M., Simonson, C., Carlso, J., Tipping, E.M., Zaleska, I., 1987. Growth promotion of canola (rapeseed) seedlings by a strain of Pseudomonas putida under gnotobiotic conditions. Canadian Journal of Microbiology 33, 390.