Subscribe Now Subscribe Today
Research Article

Evaluation of Bean (Phaseolus vulgaris) Seeds Inoculation with Rhizobium phaseoli and Plant Growth Promoting Rhizobacteria on Yield and Yield Components

M. Yadegari, H.A. Rahmani, G. Noormohammadi and A. Ayneband
Facebook Twitter Digg Reddit Linkedin StumbleUpon E-mail

To study the effect of co-inoculation with plant growth-promoting rhizobacteria (PGPR) and Rhizobium, on yield and yield components of common bean (Phaseolus vulgaris L.) cultivars was investigated in 2 consecutive years under field condition of plant growing evidence indicates that soil beneficial bacteria can positively affect symbiotic performance of rhizobia. PGPR strains Pseudomonas fluorescens P-93 and Azospirillum lipoferum S-21 as well as two highly effective Rhizobium strains were used in this study. Common bean seeds of three cultivars were inoculated with Rhizobium singly or in a combination with PGPR to evaluate their effect on growth characters. A significant variation of plant growth in response to inoculation with Rhizobium strains was observed. Treatment with PGPR significantly increased pod per plant, number of seeds per pod, weight of 100 seed, weight of seeds per plant, weight of pods per plant, total dry matter in R6 as well as seed yield and protein content. Co-inoculation with Rhizobium and PGPR demonstrated a significant increase in the yield and yield components. The results showed that all treatments of bacteria increased yield; however, strains Rb-133 with Pseudomonas fluorescens P-93 gave the highest seed yield, number of pods per plant, weight of 100 seed, seed protein yield, number seed per pod, seed protein yield.

Related Articles in ASCI
Similar Articles in this Journal
Search in Google Scholar
View Citation
Report Citation

  How to cite this article:

M. Yadegari, H.A. Rahmani, G. Noormohammadi and A. Ayneband, 2008. Evaluation of Bean (Phaseolus vulgaris) Seeds Inoculation with Rhizobium phaseoli and Plant Growth Promoting Rhizobacteria on Yield and Yield Components. Pakistan Journal of Biological Sciences, 11: 1935-1939.

DOI: 10.3923/pjbs.2008.1935.1939



The common bean (Phaseolus vulgaris L.) is an important legume for human nutrition and a major protein and calorie source in the world (Sharon, 2003). Phaseolus vulgaris can grow by assimilation on mineral nitrogen or molecular N fixation. A board range of Rhizobium species are able nodule and fix N2 with beans including R. leguminosarum biovar phaseoli, R. tropic and R. etli. The most Iran soils N deficiency, N2 fixation Rhizobium bacteria could increase yield at a low cost and preserve water resources from pollution by nitrates. It occupies more than 125,000 ha in Iran but yield remains low to moderate due to the scarce nodulation, high inputs of chemical fertilizers and low technologies applied. Common bean is usually considered a poor nitrogen-fixing legume however; its promising potential to fix nitrogen has been shown in several studies (Peix et al., 2001; Garcia et al., 2004; Remans et al., 2008). Poor nodulation and variable response to inoculation is mainly attributed to intrinsic characteristics of the host plant, particularly the nodulation promiscuity as well as the great sensitivity to other nodulation-limiting factors, such as high rates of N fertilizer used in intensive agriculture, high temperatures and soil dryness (Bais et al., 2006; Egamberdiyeva et al., 2007). Genotypic variation in beans as well as compatibility of Rhizobium-plant cultivars can also greatly affect the efficiency of symbiosis established. This variability often limits the nitrogen-fixing performance of soil native rhizobia or use of commercially available inocula. As a result, application of high amounts of inorganic nitrogen fertilizers is becoming a common practice which has detrimental environmental consequences.


Plant material and bacterial strains: Three kidney bean cultivars Sayad, Akhtar and Goli were obtained from Seeds and Plant Improvement Institute (SPII) housed at Karaj, Iran and used throughout this research. Two Rhizobium strains Rb-133 and Rb-136 with high nitrogen-fixing effectiveness were used in this study. These strains were selected during our previous screening program and demonstrated a good potential to nodulate beans and increase plant growth and yield in greenhouse and field experiments (Asadi Rahmani et al., 2005). PGPR strains Pseudomonas fluorescens P-93 and Azospirillum lipoferum S-21 positive effects in this study (Table 1). For preparation of inoculants, bacteria were grown in appropriate growth media for three days and 150 mL of each strain suspension was added to a polypropylene plastic bag containing 50 g of sterile powdered perlite and mixed thoroughly.

Table 1: Some characteristics of PGPR strains used in this study

Experimental conditions: Field trails established in 2006 and 2007 at Shahrekord (50° 51`N 32° 17`E) south western in Iran. Experiments were arranged in a randomized complete block design using a split plot layout with three replications. Seeds of common bean cultivars Sayad, Akhtar and Goli were planted after they were moistened with a 20% solution of sucrose and then inoculated (7 g inoculant per kg seed) with Rhizobium alone or in combination with P. fluorescens P-93 or A. lipoferum S-21. The experiment also included a non-inoculated control as well as one N-fertilized (100 kg N ha-1) treatments. The main-plots units consisted of 8 treatments (six bacterial combinations, one uninoculated control and one N-fertilized) applied.


We found significant different effects induced by rhizobia on growth parameters of common bean. Plant cultivars also showed different responses due to inoculation with rhizobial isolates.

Inoculation with Rhizobium significantly (p<0.01) increased number seed per pod, number pod per plant, weight of 100 seed, seed yield, weight of seed per plant, weight of pod per plant, protein seed yield and total dry matter in R6 in both years. Co-inoculation with PGPR promoted nodulation over Rhizobium alone, where the highest values for seed yield and yield components were observed by combination of Rhizobium and Pseudomonas fluorescens P-93. Similar result was obtained for shoot dry matter of plants in R6 growth stage. Rhizobium inoculation by isolates Rb-133 and Rb-136 increased dry matter production of the shoots. Co-inoculation with A. lipoferum S-21 improved dry matter production over Rhizobium treatment. This beneficial effect was greater for PGPR strain P. fluorescens P-93 and demonstrated the highest dry matter production. Application of PGPR enhanced seed yield compared to either control or Rhizobium alone. The highest seed yield was obtained from plants inoculated with Rb-133+ P. fluorescens P-93 which showed a 240% increase over control plants.

The amount of seed yield was affected by co-inoculation of Rhizobium with PGPR strains in cultivars used in this study (Table 2-4). P. fluorescens P-93 resulted in highest amount of seed yield and other characters followed by A. lipoferum S-21, when co-inoculated with Rb-133, indicating that P. fluorescens P-93 had the most promising effect on enhancement of symbiotic performance of rhizobial strains. Plants inoculated with Rhizobium alone showed different values for yield components. Plants inoculated with Rb-133+ P. fluorescens P-93 made the most seed yield, which showed over 50% increase compared to Rhizobium alone.

This research has shown the effects of co-inoculation with two PGPR strains on the symbiotic performance of common bean nodulating rhizobia in three P. vulgaris cultivars. Common bean is believed to be a poor nitrogen fixer due to the genetic characteristics of symbiotic partners as well as soil and environmental conditions. However, selecting rhizobia for increased survival in specific soil types, greater compatibility with crop species or cultivars, superior functioning under diverse climates, improved compatibility and competitiveness with other soil micro organisms and higher nitrogen-fixing efficiency have been shown that can improve growth and yield components of inoculated legumes (Vessey, 2003). Beneficial effects of rhizobia on common bean have been described in several studies with different climatic and soil conditions (Peix et al., 2001; Tilak et al., 2006; Remans et al., 2008). All plant factors we measured in the study were positively affected by inoculation with rhizobia strains Rb-133 and Rb-136. Rhizobia strains were able to increase seed yield, number of pods per plant, number of seeds per pod, weight of 100 seed, weight of seeds per plant, seed protein yield, total dry matter in R6 and protein content over uninoculated control plants. Amount of seed yield by inoculated plants was ranging from 1221 to 4693 kg ha-1 depending on the strain and cultivars used during two years of the study. However, Rb-133 showed a greater symbiotic efficiency than Rb-136. Plant cultivars also had different responses to rhizobial inoculation. Cultivar Akhtar demonstrated highest potential for seed yield, number of pods per plant, number of seeds per pod, weight of 100 seed compared to cultivars Sayad and Goli. Differences among strains of common bean rhizobia and plant cultivars in their nitrogen-fixing performance were previously observed by Vladimir (2001), Bais et al. (2006), Ahmad et al. (2006) and Remans et al. (2008). Co-inoculation of the common bean with Rhizobium and PGPR resulted in better nodulation which was translated into higher shoot dry matter and seed yield production. This is in agreement with previous reports demonstrating the beneficial effects of PGPR belonging to Pseudomonas spp. and Azospirillum spp. on symbiotic efficiency of rhizobia nodulating different legume crops (Preston, 2004; Bashan and Holguin, 2004; Valverde et al., 2006; Figueiredo et al., 2007). The results revealed that application of PGPR together with Rhizobium improved the growth and seed production by inoculated beans. As a result, gross average of seed yield increased from 1536-3000 kg ha-1 for Rhizobium alone to 1221-4693 kg ha-1 for those co-inoculated with A. lipoferum S-21 and P. fluorescens P-93, respectively. Present data showed that P. fluorescens P-93 had better promoting effect on yield components of rhizobia than A. lipoferum S-21. This difference is probably attributable to siderophore production as well as higher ability for auxin production and P-solubilizing activity of P. fluorescens P-93 (Table 1).

Table 2: Number pod per plant (PP), seed per pod (SP), weight of 100 seed (WS), weight of seed per plant (WSP), weight of pod per plant (WPP), total dry matter in R6 (TDM) and seed yield, seed protein yield by bean plants in the field experiment in the first year

Table 3: Number pod per plant (PP), seed per pod (SP), weight of 100 seed (WS), weight of seed per plant (WSP), weight of pod per plant (WPP), total dry matter in R6 (TDM) and seed yield, seed protein yield by bean plants in the field experiment in the second year

Table 4: Complex analysis of variance of number pod per plant, number seed per pod, weight of 100 seed, seed yield, weight of seed per plant, weight of pod per plant, protein seed yield, total dry matter in R6 in bean plants that affected by several bacteria treatments
ns, * and **: Non significant, significant at the 5 and 1% levels of probability, respectively


This study showed that plant growth and seed yield potential of Rhizobium-P. vulgaris varies with Rhizobium strains and plant cultivars. Co-inoculation of the common bean with Rb-133 and Pseudomonas fluorescens P-93 resulted in higher number seed per pod, weight of 100 seed, weight of seed per plant, weight of pod per plant, protein seed yield, total dry matter in R6 and thereby produced greater seed yield. The results indicate that in spite of the fact that Rb-133+ P. fluorescens P-93 can increase the proportion of seed per pod and productivity in plants, application of complementary inorganic nitrogen fertilizer in soils with low nitrogen content is needed.

1:  Ahmad, F., I. Ahmad and M.S. Khan, 2008. Screening of free-living rhizospheric bacteria for their multiple plant growth promoting activities. Microbiol. Res., 168: 173-181.
CrossRef  |  Direct Link  |  

2:  Asadi Rahmani, H., M. Afshari, K. Khavazi, F. Nourgholipour and A. Otadi, 2005. Effects of common bean nodulating rhizobia native to Iranian soils on the yield and quality of bean. Iranian J. Soil Water Sci., 19: 215-225.
Direct Link  |  

3:  Bais, H.P., T.L. Weir, L.G. Perry, S. Gilroy and J.M. Vivanco, 2006. The role of root exudates in rhizosphere interactions with plants and other organisms. Annu. Rev. Plant Biol., 57: 233-266.
CrossRef  |  PubMed  |  Direct Link  |  

4:  Egamberdiyeva, D., 2007. The effect of plant growth promoting bacteria on growth and nutrient uptake of maize in two different soils. Applied Soil Ecol., 36: 184-189.
CrossRef  |  Direct Link  |  

5:  Figueiredo, M.V.B., C.R. Martinez, H.A. Burity, C.P. Chanway, 2007. Plant growth-promoting rhizobacteria for improving nodulation and nitrogen fixation in the common bean (Phaseolus vulgaris L.). World J. Mirobiol. Biotechnol., 10.1007/s11274-007-9591-4

6:  Garcia J.A.L., A. Probanza, B. Ramos, J. Barriuso and F.J.G. Manero, 2004. Effects of inoculation with plant growth promoting rhizobacteria (PGPRs) and Sinorhizobium fredii on biological nitrogen fixation, nodulation and growth of Glycine max cv. Osumi. Plant and Soil, 267: 143-153.
Direct Link  |  

7:  Peix A., P.F. Mateos, C. Rodriguez-Barrueco, E. Martinez and E. Velazquez, 2001. Growth promotion of common bean (phaseolus vulgaris L.)by a strain of Burkholderia cepacia under growth chamber condition. Soil Biol. Biochem., 33: 1927-1935.
CrossRef  |  

8:  Preston, G.M., 2004. Plant perceptions of plant growth-promoting Pseudomonas. phil trans. R. Soc. Land. B., 359: 907-918.
CrossRef  |  

9:  Remans, R., L. Ramaekers, S. Schelkens, G. Hernandez, L. Galvez, J. Vanderleyden, 2008. Effect of Rhizobium-Azospirillum coinoculation on nitrogen fixation and yield of two contrasting Phaseolus vulgaris L. genotypes cultivated across different environments in Cuba. Plant and Soil, org/10.1007/s11104-008-9606-4

10:  Slattery, J.F., D.J. Pearce and W.J. Slattery, 2004. Effects of resident rhizobial communities and soil type on the effective nodulation of pulse legumes. Soil Biol. Biochem., 36: 1339-1346.
CrossRef  |  Direct Link  |  

11:  Tilak, K.V.B.R., N. Rannganayaki and C. Mannoharachari, 2006. Synergistic effects of plant-growth promoting rhizobacteria and Rhizobium on nodulation and nitrogen fixation by pigeon pea (Cajanus cajan). Eur. J. Soil Sci., 57: 67-71.
CrossRef  |  

12:  Valverde, A., A. Burgos, T. Fiscella, R. Rivas and E.V. Quez et al., 2006. Differential effects of coinoculations with Pseudomonas jessenii PS06 (a phosphate-solubilizing bacterium) and Mesorhizobium ciceri C-2/2 strains on the growth and seed yield of chickpea under greenhouse and field conditions. Plant Soil, 287: 43-50.
CrossRef  |  

13:  Vessey, J.K., 2003. Plant growth promoting rhizobacteria as biofertilizers. Plant Soil, 255: 571-586.
CrossRef  |  Direct Link  |  

14:  Bashan, Y., G. Holguin and L.E. de-Bashan, 2004. Azospirillum-plant relationships: Physiological, molecular, agricultural and environmental advances (1997-2003). Can. J. Microbiol., 50: 521-577.
CrossRef  |  Direct Link  |  

©  2021 Science Alert. All Rights Reserved