# Response of biofertilizers on biomass partitioning pattern and yield of pea (Pisum sativum L.)

Ajeet Kumar, Diwakar Paswan

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Ajeet Kumar, Diwakar Paswan. Response of biofertilizers on biomass partitioning pattern and yield of pea (Pisum sativum L.). AN ASIAN JOURNAL OF SOIL SCIENCE, 2014, 9 (2), pp.221-225. 10.15740/HAS/AJSS/9.2/221-225. hal-04676705

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Research Article

DOI: 10.15740/HAS/AJSS/9.2/221-225

# Response of biofertilizers on biomass partitioning pattern and yield of pea (Pisum sativum L.)

■ AJEET KUMAR AND DIWAKAR PASWAN

Received : 04.09.2014; Revised : 10.10.2014; Accepted : 27.10.2014

MEMBERS OF RESEARCH FORUM:

**Corresponding author :**

AJEET KUMAR, Regional Research
Station, Agwanpur, SAHARSA (BIHAR)
INDIA

Email: ajeetbau@gmail.com

**Co-authors :**

DIWAKAR PASWAN, Regional
Research Station, Agwanpur,
SAHARSA (BIHAR) INDIA

## Summary

Among legumes, pea (Pisum sativum L.) is the second most important grain legume, which is widely used in human nutrition. The yield potential of cultivars is one of the major factors that determine the use of vegetable pea. Currently, pre-sowing inoculation of pea seeds is a promising treatment and is one of agronomic solutions for sustainable agriculture development. The research was based on the precise field experiment, conducted in three replicates and carried out in the experimental field. The examined inoculants were applied during sowing. The presented results of the studies on the symbiotic nitrogen fixation and phosphate solubilizers by leguminous plants indicate that the productivity of pea was positively affected by the application of microbial inoculants. The supplementation of live inoculums of Rhizobium, Azotobacter, Azospirillum and phosphate solubilizing bacteria (PSB) in the rhizosphere of pea can significantly enhanced the biomass accumulation and yield (27 – 55 %) as compared to uninoculated control. The degree of influence was found to be dependent on variety and preference of genotypes for biofertilizer. A steady increase in root, shoot and leaf biomass accumulation from seedling stage to flowering was evidenced in all the three variety. The shredding of leaf biomass was highest irrespective of varieties followed by shoot biomass at pod formation stage, thereafter up to senescence the leaf and shoot biomass was static. A preference to PSB for arkel and rhizobium plus free N fixers for Azad P₁ and Azad P₂ in terms of accumulation of partitioned biomass were noticed which suggested the use of low cost biofertilizers for higher productivity in pea.

**Key words:** Biofertilizers, Pea varieties, Biomass, Yield

**How to cite this article:** Kumar, Ajeet and Paswan, Diwakar (2014). Response of biofertilizers on biomass partitioning pattern and yield of pea (Pisum sativum L.). *Asian J. Soil Sci.*, 9(2): 221-225.

## Introduction

Grain legumes are widely recognized as a significant source of proteins (Duranti and Guis, 1997) and have become very important in human and domestic animal nutrition (Cummings *et al.*, 2001). Presently, the increasing interest in low input sustainable agricultural systems on cropping farms, opens the door for continued interest in grain legumes (White, 1989). Among legumes, pea (Pisum sativum L.) is the second most important grain legume crop, which is widely used in human nutrition and as fodder (Cristou, 1997). The importance of legumes to these purposes is not only due to their nitrogen (N) fixing capabilities but also due to their ability to interrupt the cycles of diseases and pests affecting

other crops (Cummings *et al.*, 2001). Many authors claimed that yields could be increased by legume seed inoculation compared to nitrogen fertilizer variants (Dobereiner *et al.*, 1995). Legume inoculation is an established practice to introduce rhizobia into the soil (Deaker *et al.*, 2004). Symbiotic effectiveness is one of the most important factors when selecting an inoculant strain (Stephens and Rask, 2000). Inoculants can be applied directly onto seeds, which is the traditional and most commonly used method of inoculation (Wadoux, 1991). Inoculation of legumes has produced variable results when different inoculation methods were used. An alternative method is the application of the inoculants to the soil at the time of sowing. Soil application of the inoculum has been successful in field pea (Pisum
*sativum* L.). This study was conducted to evaluate the productivity of three different pea varieties in order to assess the variability of morphological traits and yield components after inoculation with symbiotic bacteria (*Rhizobium*).

Global demand for food will increase commensurately with a world population that may grow to reach 8.3 billion by 2025 (Sylvia *et al.*, 2005). Such increased demand will most likely occur in developing countries like India, many of which suffer from limited access to fertilizers and other exogenous farm inputs (Sylvia *et al.*, 2005). The symbiotic association between *Rhizobium* sp. and pea (*Pisum sativum*) is well documented in the literature. Rhizobia-mediated biological N-fixation, which occurs on legume roots, can provide residual fertilizer to subsequent small grain crops. Specifically, rhizobiaceae are free-living diazotrophic saprophytes able to form nitrogen- (N-) fixed symbiotic associations with legumes by forming root nodules. Pea plants (*Pisum sativum* L.) are nodulated by *Rhizobium leguminosarum* bv. *viciae* (Jordan, 1984). In the recent years the application of free N fixers like *Azotobacter*, *Acetobacter* and *Azospirillum* coupled with Phosphate Solubilizing Bacteria in the rhizosphere are gaining importance. Scientific literatures indicated the significant positive effect of these microorganisms on peas and beans besides cereals and sugarcane crops (Yakout *et al.*, 2001 and Wu *et al.*, 2005). Dry matter synthesis, accumulation and their distribution pattern are dependent on the genotypic characteristics of plants and environmental factors influence the rhizosphere chemistry and biological equilibrium of soil (Wu *et al.*, 2005; Tiwary *et al.*, 1999 and Srivastava *et al.*, 1998). Studies on the biomass production and its partitioning are important aspect of crop management because grain yield largely depended on the partitioning of photosynthates at critical crop growth stages (Clark and Smith, 1992 and Yan, 1981). The present experiment was carried out with *Rhizobium*, *Azotobacter*, *Azospirillum* and PSB and observed their impact on the biomass-partitioning pattern of three vegetable pea varieties. This study reports on biomass partitioning in vegetable pea sequentially over three growing phase covering the period from active growth stage to maturity and senescence.

## Resource and Research Methods

An explorative trial on impact of biofertilizers on vegetable pea varieties was conducted during *Rabi* season (November, 2008 to January, 2009). The experiment was repeated during *Rabi* season in 2009-2010 and 2010-2011 at the regional research farm, Agwanpur (86.34° longitudes and 25.55° N latitude), Saharsa, Bihar. The experiment was conducted with three most promising varieties of pea (Azad $P_1$, Azad $P_3$ and Arkel) and four biofertilizers *Rhizobium*, *Azotobacter*, *Azospirillum* and Phosphate Solubilizing

Bacteria along with control (no applied biofertilizers) in a randomized block design (RBD) with three replications. The soil of the experimental site was sandy loam in texture, neutral in reaction (pH 7.2-7.8), low to medium in available N (275-310 kg/ha) and K (290-340 kg/ha) and medium to high in available P (15-25 kg/ha). The physico-chemical properties of the soil were analyzed, following the standard procedure Jackson (1967). Certified seed of pea variety were treated @ 500 g/10 kg seeds of individual N fixing biofertilizers following the standard methodology, and sown in each plot at a spacing of 30 x 10 cm with the mechanical seed drill and then covered with PSB @ 25 kg/ha plus N fixer @ 10 kg/ha mixed with finely tilth garden soil. The recommended fertilizer doses N @ 30 kg/ha, P @ 60 kg/ha and K @ 60 kg/ha were applied during the field preparation. The plants were sampled at active growth stage ($S_1$ = 20 days after sowing, DAS), pod-filling stage ($S_2$, $S_3$) and at maturity/harvesting stage (75 DAS, $S_3$) for the observation of partitioned biomass. On each sampling occasion, the biomass was separated into leaf, stem, and root, weighed and oven dried (60°C) for further chemical analysis. The yield and yield attributing parameters were recorded and correlated with the biomass-partitioning pattern.

## Research Findings and Discussion

The application of microbial inoculants as seed treatments and soil application were influenced significantly the accumulated biomass in fresh leaf, shoot and roots of vegetable pea at three critical growth stages. The highest amount of fresh leaf biomass (29.19, 79.43 and 59.32 g) accumulated at $S_1$, $S_2$ and $S_3$ growth stages, respectively was observed in Azad $P_3$ when treated with *Rhizobium* (Table 1). The fresh leaf biomass was 60.3 per cent, 63.42 per cent and 66.2 per cent higher when treated with *Azotobacter*, *Rhizobium* and PSB, respectively as compared to the control treatment in cv. Arkel. The maximum leaf biomass accumulation stage coincided with the pod filling stage (45 DAS) in all the three varieties. The reduction in the peak of leaf biomass production was observed in all the three variety of pea just after the pod filling stage. The overall performance of Azad $P_3$ was significantly superior as compared to Arkel and Azad $P_1$ in relation to leaf biomass production. The highest average fresh leaf weight (24.27 and 63.19 g) at active and pod filling stage in Azad $P_1$ was recorded when treated with PSM and *Azotobacter*, respectively, which was 59 per cent and 68.85 per cent higher than the control pea. In Arkel the highest average fresh leaf weight (28.97 and 76.44 g) at active and pod filling stage was observed when treated with *Azotobacter* and *Azospirillum*, respectively which was 55.3 per cent and 45.18 per cent higher than the control pea. The maximum shoot biomass production (89.82 g) was achieved at pod filling stage by Azad $P_3$ cultivar when treated with

<figure><img src="image_3.png" /></figure>
*Rhizobium* strains that was 61.95 per cent higher than that of control pea. The bacterium *Rhizobium leguminosarum* lives symbiotically in the roots of leguminous plants. It infects roots through root hairs of pulse crops like sweet pea, lentil, garden pea, broad bean, lobia, etc. and reproduce in the

cortical cells of the root. With simultaneous divisions of cortex cells nodules are formed in the root. The bacterium *Rhizobium* converts the absorbed atmosphere nitrogen into ammonia, which is further converted into other nitrogenous substances. Rhizobium fixes almost ten times more nitrogen than it uses.

<table><caption>Table 1: Effect of biofertilizers on fresh biomass weight (g) of three vegetable pea varieties</caption><thead><tr><th rowspan="3">Treatments of biofertilizers</th><th rowspan="3">Biomass</th><th colspan="3">Azad P<sub>1</sub></th><th colspan="3">Azad P<sub>3</sub></th><th colspan="3">Arkel</th></tr><tr><th colspan="3">Different growth stages</th><th colspan="3">Different growth stages</th><th colspan="3">Different growth stages</th></tr><tr><th>S<sub>1</sub></th><th>S<sub>2</sub></th><th>S<sub>3</sub></th><th>S<sub>1</sub></th><th>S<sub>2</sub></th><th>S<sub>3</sub></th><th>S<sub>1</sub></th><th>S<sub>2</sub></th><th>S<sub>3</sub></th></tr></thead><tbody><tr><td rowspan="3">Azotobacter</td><td>Leaf</td><td>23.1</td><td>63.19</td><td>43.21</td><td>26.12</td><td>69.88</td><td>48.43</td><td>28.97</td><td>70.3</td><td>63.21</td></tr><tr><td>Shoot</td><td>24.93</td><td>62.52</td><td>55.29</td><td>41.92</td><td>75.79</td><td>51.33</td><td>33.92</td><td>75.82</td><td>65.92</td></tr><tr><td>Root</td><td>1.43</td><td>2.18</td><td>2.15</td><td>1.46</td><td>1.69</td><td>1.70</td><td>1.43</td><td>1.56</td><td>1.51</td></tr><tr><td rowspan="3">PSB</td><td>Leaf</td><td>24.27</td><td>62.99</td><td>45.92</td><td>27.76</td><td>78.43</td><td>59.32</td><td>28.77</td><td>69.37</td><td>54.38</td></tr><tr><td>Shoot</td><td>29.25</td><td>68.91</td><td>49.30</td><td>45.31</td><td>83.18</td><td>63.28</td><td>35.19</td><td>73.37</td><td>58.53</td></tr><tr><td>Root</td><td>1.45</td><td>2.42</td><td>2.28</td><td>1.47</td><td>1.69</td><td>1.73</td><td>1.46</td><td>1.70</td><td>1.71</td></tr><tr><td rowspan="3">Rhizobium</td><td>Leaf</td><td>23.91</td><td>41.05</td><td>35.78</td><td>29.19</td><td>79.43</td><td>52.28</td><td>28.73</td><td>68.52</td><td>51.58</td></tr><tr><td>Shoot</td><td>34.97</td><td>53.29</td><td>38.18</td><td>59.63</td><td>89.82</td><td>55.91</td><td>37.26</td><td>71.53</td><td>55.39</td></tr><tr><td>Root</td><td>1.41</td><td>1.59</td><td>1.53</td><td>1.41</td><td>1.37</td><td>2.53</td><td>1.4</td><td>1.89</td><td>1.92</td></tr><tr><td rowspan="3">Azospirillum</td><td>Leaf</td><td>24.07</td><td>43.51</td><td>29.31</td><td>26.88</td><td>72.08</td><td>45.93</td><td>28.63</td><td>76.44</td><td>65.32</td></tr><tr><td>Shoot</td><td>31.32</td><td>49.31</td><td>31.73</td><td>55.36</td><td>75.52</td><td>49.89</td><td>39.45</td><td>79.43</td><td>53.31</td></tr><tr><td>Root</td><td>1.34</td><td>1.26</td><td>1.29</td><td>1.50</td><td>2.59</td><td>1.39</td><td>1.32</td><td>1.68</td><td>1.69</td></tr><tr><td rowspan="3">Control</td><td>Leaf</td><td>14.32</td><td>35.27</td><td>27.32</td><td>17.61</td><td>50.38</td><td>39.31</td><td>16.03</td><td>34.54</td><td>22.38</td></tr><tr><td>Shoot</td><td>17.92</td><td>37.28</td><td>29.39</td><td>21.49</td><td>55.65</td><td>42.99</td><td>21.90</td><td>21.38</td><td>29.98</td></tr><tr><td>Root</td><td>0.85</td><td>1.51</td><td>1.53</td><td>0.81</td><td>1.43</td><td>1.21</td><td>0.76</td><td>1.09</td><td>1.01</td></tr><tr><td rowspan="3">LSD<sub>0.05</sub></td><td>Leaf</td><td>0.69</td><td>1.23</td><td>2.93</td><td>0.69</td><td>1.23</td><td>2.93</td><td>0.69</td><td>1.23</td><td>2.93</td></tr><tr><td>Shoot</td><td>5.69</td><td>8.23</td><td>3.93</td><td>5.69</td><td>8.23</td><td>3.93</td><td>5.69</td><td>8.23</td><td>3.93</td></tr><tr><td>Root</td><td>0.09</td><td>0.12</td><td>0.35</td><td>0.09</td><td>0.12</td><td>0.35</td><td>0.09</td><td>0.12</td><td>0.35</td></tr></tbody></table>

S<sub>1</sub>-active growth stage, S<sub>2</sub>-pod filling stage, S<sub>3</sub>-harvesting stage

<table><caption>Table 2: Effect of biofertilizers on pod number/plant (C<sub>1</sub>), pod length (C<sub>2</sub>), pod diameter (C<sub>3</sub>), pod weight (C<sub>4</sub>), number of grain/pod (C<sub>5</sub>) and yield of vegetable pea</caption><thead><tr><th>Treatments of Biofertilizers</th><th>Pea varieties</th><th>C<sub>1</sub></th><th>C<sub>2</sub> (cm)</th><th>C<sub>3</sub> (cm)</th><th>C<sub>4</sub> (g)</th><th>C<sub>5</sub></th><th>Yield (Q/ha)</th></tr></thead><tbody><tr><td rowspan="3">Azotobacter</td><td>Azad P<sub>1</sub></td><td>8.56</td><td>7.43</td><td>3.47</td><td>3.1</td><td>6.72</td><td>78.46</td></tr><tr><td>Azad P<sub>3</sub></td><td>8.24</td><td>6.38</td><td>3.51</td><td>3.2</td><td>5.8</td><td>85.39</td></tr><tr><td>Arkel</td><td>8.90</td><td>6.80</td><td>3.48</td><td>2.9</td><td>5.37</td><td>71.18</td></tr><tr><td rowspan="3">PSB</td><td>Azad P<sub>1</sub></td><td>7.62</td><td>6.60</td><td>3.22</td><td>2.7</td><td>6.7</td><td>61.50</td></tr><tr><td>Azad P<sub>3</sub></td><td>8.70</td><td>7.20</td><td>3.92</td><td>3.0</td><td>5.43</td><td>77.33</td></tr><tr><td>Arkel</td><td>9.3</td><td>6.68</td><td>3.35</td><td>2.8</td><td>5.23</td><td>83.46</td></tr><tr><td rowspan="3">Rhizobium</td><td>Azad P<sub>1</sub></td><td>6.86</td><td>7.07</td><td>3.54</td><td>3.5</td><td>6.5</td><td>67.28</td></tr><tr><td>Azad P<sub>3</sub></td><td>7.5</td><td>6.74</td><td>3.43</td><td>3.2</td><td>5.97</td><td>70.37</td></tr><tr><td>Arkel</td><td>8.22</td><td>6.67</td><td>3.40</td><td>2.8</td><td>7.0</td><td>68.11</td></tr><tr><td rowspan="3">Azospirillum</td><td>Azad P<sub>1</sub></td><td>8.73</td><td>7.02</td><td>3.37</td><td>3.5</td><td>7.37</td><td>80.59</td></tr><tr><td>Azad P<sub>3</sub></td><td>9.8</td><td>7.74</td><td>3.42</td><td>4.0</td><td>5.8</td><td>92.63</td></tr><tr><td>Arkel</td><td>8.35</td><td>7.31</td><td>3.27</td><td>2.9</td><td>5.33</td><td>78.96</td></tr><tr><td rowspan="3">Control</td><td>Azad P<sub>1</sub></td><td>7.66</td><td>6.57</td><td>3.33</td><td>3.0</td><td>6.47</td><td>52.14</td></tr><tr><td>Azad P<sub>3</sub></td><td>6.07</td><td>6.75</td><td>3.47</td><td>3.8</td><td>5.6</td><td>58.22</td></tr><tr><td>Arkel</td><td>6.75</td><td>6.90</td><td>3.45</td><td>3.1</td><td>5.43</td><td>53.27</td></tr><tr><td rowspan="3">LSD (0.05)</td><td>Azad P<sub>1</sub></td><td>0.93</td><td>0.21</td><td>0.26</td><td>0.3</td><td>0.48</td><td>4.09</td></tr><tr><td>Azad P<sub>3</sub></td><td>0.93</td><td>0.21</td><td>0.26</td><td>0.30</td><td>0.48</td><td>4.09</td></tr><tr><td>Arkel</td><td>0.93</td><td>0.21</td><td>0.26</td><td>0.30</td><td>0.48</td><td>4.09</td></tr></tbody></table>

<figure><img src="image_4.png" /></figure>
Findings show similarities with Sarkar (2006) who also observed that *Rhizobium* population was substantially increased with the application of organic wastes. The positive effects of these treatments were also observed with the increase in both total and effective nodules per plant in peanut crop. The reason for the improved biological nitrogen fixation (BNF) is most likely a combination of factors related to availability of nutrients in the soil (Lehmann *et al.*, 2003) and stimulation of plant-microbe interactions (Nishio and Okano, 1991; Saito and Marumoto, 2002). The root and shoot biomass almost remain constant while leaf biomass decreased significantly from pod filling stage to maturity and senescence. The decrease in the leaf biomass was partitioned towards the formation of pod. Similar trend in shredding leaf biomass for pod formation in other crops has been reported (Hay, 1995 and Zhang *et al.* 2004). The highest amount of average fresh root biomass (2.42 g) in Azad $P_1$ at pod filling stage was observed when treated with PSB. The PSB mobilises fixed/non-exchangeable P to readily available labile P which helped in root proliferation and nodulation of pea. The PSB resulted in significant increases in nodulation, nitrogenase activity, growth and grain yield of pea (Srivastava *et al.*, 1998). Similarly the highest fresh root biomass (1.47 g) was recorded in Azad $P_3$ (inoculated with PSB) at active growth stage that was 55.1 per cent higher than the control pea. The highest fresh root weight (2.59 g) was observed at pod filling stage when treated with *Azospirillum*, which was 55.2 per cent higher than the control pea. In Arkel the highest average fresh root weight (1.43 g) was noted at active growth stage when treated with *Azotobacter* followed by *Azospirillum* at pod setting stage which was 53.15 per cent and 60.8 per cent higher than the control pea. The highest value of mean leaf dry weight (14.86 g) among all the three pea cultivars was observed in *Rhizobium* treated Azad $P_3$ which was 41.79 per cent higher than the control pea. The highest mean shoot and root dry weight (17.82 and 1.34 g) again reported with the Azad $P_3$ treated with PSM and *Azospirillum* inoculants, which was 89.88 per cent and 66.41 per cent higher than, uninoculated pea, respectively. The inoculation of *Rhizobium* sp. and free N - fixers induced fixation of more amount of atmospheric nitrogen in the active root nodule of vegetable pea, increased leaf area index, which increases the utilization of solar energy and enhanced the accumulation of photosynthates in the tissue (Lecoeur and Ney, 2003; Aira and Paivoke, 2003). Increased dry matter production in the senna (*Cassia angustifolia* Vahl.) had been reported when the seeds were treated with *Azospirillum* (Arumugam *et al.*, 2001).

The number of pods per cluster as well as number of pods per plant was enhanced significantly due to the effect of biofertilizers. The highest number of pods per cluster (2.13) was recorded in Azad $P_1$ when inoculated with the *Azospirillum*, which was 71.83 per cent higher to control.

The highest average number of pods/plant (9.8) was reported in Azad $P_3$ inoculated with *Azospirillum* (45.05 % more than control). The response of Arkel was very impressive towards the application of PSB in relation to number of pod/plant (9.3) and number of pods per cluster. The highest pod length (7.74) was recorded in Azad $P_3$ (inoculated with *Azospirillum*) that was 14.6 per cent higher to control where as highest pod diameter (3.92) was reported in the same varieties inoculated with the PSB, 12.9 per cent higher to control (Table 2). The highest average pod weight (4.0 g) was recorded in Azad $P_3$ inoculated with the *Azospirillum* which was 5.2 per cent higher to uninoculated plant. The highest number of grains per pod (7.37) was recorded in Azad $P_1$ (inoculated with *Azospirillum*), which was 13.9 per cent higher to the control pea. Green pod yield was significantly higher in Azad $P_3$ (92.63 q/ha) due to applications of *Azospirillum*, which was 55 per cent higher to control pea. The symbiotic association of Azad $P_3$ to *Rhizobium/Azospirillum*, Azad $P_1$ to *Azospirillum/Azotobacter* and Arkel to PSB was more evident. The present finding leads to the understanding that the extraneous application of inoculants of N fixer and PSB in the rhizosphere of pea significantly enhanced the yield and yield attributes. The application of *Azospirillum/Azotobacter* and PSB together with *Rhizobium* may be advocated for higher biomass and yield in pea.

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9th
Year

★★★★★of Excellence★★★★