Int.J.Curr.Microbiol.App.

Sci (2021) 10(01): 1588-1598

International Journal of Current Microbiology and Applied Sciences

ISSN: 2319-7706 Volume 10 Number 01 (2021)
Journal homepage: http://www.ijcmas.com

Original Research Article https://doi.org/10.20546/ijcmas.2021.1001.185

Effect of Rhizobium Inoculation Methods on Growth, Nodulation

and Yield of Black Gram (Vigna mungo L.)

Shubha Tripathi, Satendra Kumar*, Mukesh Kumar, Ashok Kumar,

B. P. Dhyani and Yogesh Kumar

1
Department of Soil Science and Agricultural Chemistry, 2Department of Agronomy, Sardar
Vallabbhai Patel University of Agriculture and Technology, Meerut, (UP), India
*Corresponding author

ABSTRACT

A field experiment was conducted to assess the effect of Rhizobium inoculation on

nutrient uptake and productivity at Crop Research Centre of Sardar Vallabhbhai Patel
Keywords University of Agriculture and Technology, Meerut (U.P.), during kharif season of
2019.The experiment was arranged in randomized block design with nine treatments in
Black gram,
three replications. Treatment consisted of T 1 (control), T2 (RDF), T3 (RDF + FYM), T4
Rhizobium
(RDF + Vermicompost), T5 (RDF + Rhizobium seed inoculation), T 6 (RDF + Rhizobium
soil treatment), T7 (RDF + Rhizobium inoculation with Vermicompost), T 8 (RDF +
Article Info
Rhizobium seed inoculation + Rhizobium soil inoculation), T 9 (RDF + Rhizobium seed
inoculation + Rhizobium inoculation with Vermicompost. From the data , it was observed
Accepted: that RDF + Rhizobium seed inoculation + Rhizobium inoculation with vermicompost
12 December 2020
Available Online:
treatment exhibited significant influence on the growth, nodulation, yield attributes and
10 January 2021 yields, of black gram as compared to control during the year of experimentation with
higher values for all the above mentioned parameters and also 90.82 and 41.68 % increase
in grains and straw yield.

Introduction production comes from India, of which black

gram constitutes 1.65 Mt with the share of
After cereals and oilseeds, pulses occupy an 12.4 % (Elzebroek and Wind, 2008). Pulses
important place in Indian agriculture. The are excellent source of high quality protein,
total world acreage under pulses is about essential amino acids, fatty acids, fibers,
85.40 Mha with production of 87.40 Mt at minerals and vitamins. The protein level of
productivity 1023 kg ha-1 production with 34 black gram is quite high i.e about 24 % with
and 26 % respectively with average nutritional value of 10.9 % moisture, 1.4 %
productivity of 835 kg ha-1 (Agricultural fats, 60.3 % carbohydrates and 3.4 % ash
Statistics Division, Directorate of Economics (Shroti et al., 2018). It improves soil health by
and Statistics, 2019). 70 % of the total enriching nitrogen status and also maintains
world’s black gram and green gram sustainability of the cropping systems. Most

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of its nitrogen requirement is fulfilled by due to less number of Rhizobia and

symbiotic nitrogen fixation from air and ineffective native Rhizobia, nodulation and
substantial amount of residual nitrogen and nitrogen fixation does not occur properly. To
organic matter are left behind for subsequent ensure efficient population of effective
crops. These crops do not have much Rhizobia in soil, these are introduced in soil
nutritional requirements and these from outside in the form of biofertilizers.
requirements can be fulfilled using various Seed treatment as well as soil application is
organic manures and bio-fertilizers which the way to apply biofertilizer. Another
prove to be as cheapest and environmentally approach may be application of biofertilizer
sound practice of applying nutrition to soil through organic sources.
and plant.
Materials and Methods
Vermicompost have been effective in
promoting the development of beneficial The field experiment was conducted in kharif
organisms in the soil. It enhances the growth, season of 2019 at Crop Research Centre,
yield and quality of crops (Meena 2013; Sardar Vallabhbhai Patel University of
Mujahid and Gupta, 2010) and also increases Agriculture and Technology, Meerut (U.P.),
bio-fertilizers efficiency. FYM through its India, to evaluate the “Effect of Rhizobium
positive effects on soil physical, chemical and inoculation methods on root nodulation,
biological properties and balanced plant nutrient uptake and productivity of Black
nutrition plays an important role in improving gram (Vigna mungo L. Hepper)”. The
the fertility and productivity of soils (Kumar experiment was arranged in randomized block
et al., 2011). Although, it cannot be design with nine treatments in three
considered as a major source of nutrients but replications, each plot size being 3.0m x
can be regarded as a good complimentary and 4.0m. Treatment consisted of T1 (control), T2
supplementary source with mineral or (RDF), T3 (RDF + FYM), T4 (RDF +
chemical fertilizers. Use of organic source of Vermicompost), T5 (RDF + Rhizobium seed
nutrient is majorly appreciated over any inoculation), T6 (RDF + Rhizobium soil
chemical fertilizer due to the fact that they are treatment), T7 (RDF + Rhizobium inoculation
not involved in causing any type of pollution with Vermicompost), T8 (RDF + Rhizobium
and they cut those higher costs of cultivation seed inoculation + Rhizobium soil
that are caused by using mineral fertilizers. inoculation), T9 (RDF + Rhizobium seed
inoculation + Rhizobium inoculation with
Black gram, being leguminous crop, use Vermicompost). Soil collected from research
symbiotic nitrogen fixation which is farm was analysed by standard procedure for
performed with the help of bacterium called various initial physicochemical properties
Rhizobia as an important source to fulfil its given in parentheses, viz. bulk density (1.38 g
major part of nitrogen requirement (Pareek, cm-3), particle density (2.65 g cm-3), pH (8.2),
1978).This symbiotic nitrogen fixation proves EC (0.28 dSm-1), organic carbon (0.45%),
to an important source of nitrogen, and porosity (46.52%), available N (150.0 kg ha-
around 200 to 300 kg nitrogen ha-1is often 1
), available P (11.30 kg ha-1) and available K
fixed by various legume crops and pasture (170.0 kg ha-1).For preparing an experimental
species (Dudeja and Duhan, 2005). These field 1 deep ploughing and 2 harrowing was
microbes vary in number, effectiveness and done and pre sowing irrigation was given at
nodulation. Although Rhizobia is great source least 10 days before sowing.
of symbiotic nitrogen fixation but sometimes

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Recommended dose of NPK (20:60:00) was (18.1, 43.3, 46.0 cm, respectively). Similar
applied through urea, single superphosphate. results were found for number of branches
Also FYM and Vermicompost were applied plant-1, number of trifoliate leaf plant-1 and
in the field @ 20-25 tonnes ha-1 and 5 tonnes dry matter accumulation plant-1 with higher
ha-1 respectively. Urad-10B strain of values as 3.83, 4.92 and 5.96, 3.9, 9.6, 8.5,
Rhizobium leguminosarum var. phaseoli and 7.10, 14.06, 14.50 g plant-1 respectively at
brought from IARI, New Delhi was used to 30, 60 and harvest stage.
treat seed, vermicompost and soil. For 10 kg
seed 200g of Rhizobium culture was used to Significant increase in plant height was found
inoculate the seed. For this, 5% solution of due to Rhizobium inoculation, which may
jaggery in water was prepared. maintain favourable balance between the
applied nutrients in the plant for its optimum
The water was taken in amount which is growth while elongation and chlorophyll
enough to moisten the seed. In this solution biosynthesis in turn, improve the branches
one packet of Rhizobium culture was added plant-1. The result is supported by Singh and
and mixed well. The coating of seed with Pareek (2003), Sripriya et al., (2005), Kumar
prepared slurry was done. Then the seeds and Elamathi (2007) Bhuiyan et al., (2008)
were dried in shade and sown immediately. and Giri et al., (2010).
Similarly, manual mixing of Rhizobium with
Vermicompost, FYM or for its incorporation Nodulation
in soil was done. The black gram variety
Shekhar-2 was sown at the rate of 15 kg ha- The number of nodules plant-1 and their dry
1
.Intercultural operations viz., weeding, weight in black gram was significantly
irrigation and insecticide spray were done as influenced by Rhizobium inoculation methods
and when required. The yield, nutrient content (Table-2 and Fig. 2). The highest number of
and uptake and physico-chemical properties nodules and their dry weight 41.0 plant-1 and
were recorded at pertinent stages. 50.10 mg plant-1, respectively were recorded
in T9 (RDF + Rhizobium seed inoculation +
Results and Discussion Rhizobium inoculation with Vermicompost) at
45 DAS. However, lowest number of nodules
Growth parameters and their dry weight 24.3 plant-1 and 34.18
mg plant-1 respectively were recorded in
In the present study Rhizobium inoculation control (T1).
methods exhibited a significant effect on
various growth parameters viz. plant height, It might have resulted due to more
number of branches and dry matter competitive ability of microbes near roots
accumulation plant-1 recorded at different which is the site for microbial infection. Well-
time periods of crop growth (Table-1 and Fig. developed root system provides more
1.1, 1.2, 1.3). At 30 and 60 DAS and at evidence for infection resulting in greater
harvest stage the treatment T9 (RDF + number of nodules.
Rhizobium seed inoculation + Rhizobium
inoculation with Vermicompost) shows These finding are found relevant to Hussain et
significantly higher plant height (23.6, 54.5, al., (2015), Dhakal et al., (2016), Meena and
57.8 cm) than the rest of the treatments. The Ram (2016), Kant et al., (2016) and
shortest plant height was obtained in control Mohammad et al., (2017).
plot at all the time period of crop growth

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Table.1 Effect of Rhizobium inoculation methods on growth parameters of black gram

Treatments Plant height(cm) No. of branch plant-1 No. of trifoliate leaf plant- Dry matter accumulation
1
(g plant-1)
30 60 At 30 60 At 30 60 At 30 60 At
DAS DAS harvest DAS DAS harvest DAS DAS harvest DAS DAS harvest
T1 – Control 18.1 43.3 46.0 2.14 3.25 4.41 1.9 6.3 5.2 5.54 8.78 9.50
T2 – RDF 20.0 46.3 51.2 2.42 3.64 4.78 2.1 7.7 6.6 5.82 9.90 11.20
T3 - RDF + FYM 20.1 47.5 51.9 2.66 3.79 5.19 2.3 7.9 6.8 5.96 10.20 12.55
T4 - RDF + 21.1 49.6 53.0 2.82 3.90 5.26 2.7 8.5 7.2 6.34 11.26 12.30
Vermicompost
T5 - RDF + Rhizobium 21.9 52.1 55.7 3.28 4.57 5.69 3.4 9.3 8.2 6.70 12.84 13.65
seed inoculation
T6 - RDF + Rhizobium 21.4 49.8 53.9 2.98 4.21 5.45 2.8 8.7 7.4 6.51 11.68 12.90
soil treatment
T7 - RDF + Rhizobium 21.6 51.0 54.7 3.10 4.36 5.57 3.2 9.1 7.9 6.53 12.15 13.50
inoculation with
Vermicompost
T8 - RDF + Rhizobium 22.5 53.3 56.3 3.58 4.78 5.82 3.6 9.4 8.3 6.88 13.58 13.68
seed inoculation +
Rhizobium soil
inoculation
T9 - RDF + Rhizobium 23.6 54.5 57.8 3.83 4.92 5.96 3.9 9.6 8.5 7.10 14.06 14.50
seed inoculation +
Rhizobium inoculation
with Vermicompost
SEm ± 0.7 1.5 1.8 0.14 0.18 0.26 0.10 0.30 0.30 0.17 0.38 0.49
CD (P= 0.05) 1.9 4.3 5.2 0.39 0.52 0.75 0.30 0.90 0.88 0.48 1.10 1.44

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Table.2 Effect of Rhizobium inoculation methods on nodulation of black gram

Treatment Number of root nodules plant-1 Dry weight of nodules plant-1 (mg plant-1)
45 DAS 60 DAS 45 DAS 60 DAS
T1 – Control 24.3 22.2 34.18 28.22
T2 – RDF 30.8 26.0 42.36 33.71
T3 - RDF + FYM 32.1 29.7 43.38 35.16
T4 - RDF + Vermicompost 34.0 31.1 45.12 37.62
T5 - RDF + Rhizobium seed inoculation 38.6 35.4 47.56 39.41
T6 - RDF + Rhizobium soil treatment 36.2 32.0 45.41 37.72
T7 - RDF + Rhizobium inoculation with 37.3 33.3 46.34 38.28
Vermicompost
T8 - RDF + Rhizobium seed inoculation + 39.2 36.2 48.58 40.38
Rhizobium soil inoculation
T9 - RDF + Rhizobium seed inoculation + 41.0 38.6 50.10 42.67
Rhizobium inoculation with Vermicompost
SEm ± 0.48 0.43 0.64 0.58

CD (P= 0.05) 1.42 1.30 1.89 1.72

Table.3 Effect of Rhizobium inoculation methods on yield attributes of black gram

Treatment Pod length (cm) No. of pod plant-1 No. of grain pod-1 Test weight (g)
T1 – Control 4.0 18.30 4.09 29.25
T2 – RDF 4.6 19.10 4.60 31.26
T3 - RDF + FYM 4.8 19.70 4.82 32.28
T4 - RDF + Vermicompost 5.1 20.65 5.17 32.68
T5 - RDF + Rhizobium seed inoculation 6.1 23.10 5.58 34.88
T6 - RDF + Rhizobium soil treatment 5.5 21.72 5.32 33.21
T7 - RDF + Rhizobium inoculation with Vermicompost 5.8 22.14 5.41 33.72
T8 - RDF + Rhizobium seed inoculation + Rhizobium soil 6.5 24.63 5.74 35.02
inoculation
T9 - RDF + Rhizobium seed inoculation + Rhizobium 6.9 25.68 5.95 35.42
inoculation with Vermicompost
SEm ± 0.32 0.51 0.25 0.44
CD (P= 0.05) 0.92 1.49 0.72 NS

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Table.4 Effect of Rhizobium inoculation methods on grain, straw, biological yield (q ha-1) and harvest index (%) of black gram

Treatment Yield (q ha-1) Harvestindex

Grain Straw Biological (%)
Yield Yield Yield
T1 – Control 6.21 21.29 29.17 21.29
T2 – RDF 8.60 21.98 39.12 21.98
T3 - RDF + FYM 9.75 23.73 41.08 23.73
T4 - RDF + Vermicompost 9.90 23.96 41.32 23.96
T5 - RDF + Rhizobium seed inoculation 10.92 25.32 43.13 25.32
T6 - RDF + Rhizobium soil treatment 10.05 23.93 42.00 23.93
T7 - RDF + Rhizobium inoculation withVermicompost 10.81 25.17 42.94 25.17
T8 - RDF + Rhizobium seed inoculation + Rhizobium soil 11.43 26.07 43.84 26.07
inoculation
T9 - RDF + Rhizobium seed inoculation + Rhizobium 11.85 26.70 44.38 26.70
inoculation with Vermicompost
SEm ± 0.33 0.81 1.37 0.81
CD (P= 0.05) 0.98 2.34 4.00 2.34

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Fig.1.1 Effect of Rhizobium inoculation methods on plant height (cm) of black gram plant-1 black
gram at different growth periods

Fig.1.2 Effect of Rhizobium inoculation methods on number of branches plant-1 at different

growth periods

Fig.1.3 Effect of Rhizobium inoculation methods on number of trifoliate leaf plant-1 (left) and dry
matter accumulation (g plant-1) (right) of black gram at different growth periods

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Fig.2 Effect of Rhizobium inoculation methods on number of nodules plant-1 and dry weight of
nodules plant-1 (mg plant-1) at different growth periods of black gram

Fig.3 Effect of Rhizobium inoculation methods on yield attributes of black gram

Fig.4 Effect of Rhizobium inoculation methods on grain, straw, biological yield (q ha-1) and
harvest index (%) of black gram

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Yield and Yield Attributes 43.84 q ha-1 in T8 (RDF + Rhizobium seed

inoculation + Rhizobium soil treatment)
Yield attributes while, lowest 22.96 and 29.17 q ha-1 was
found in control (T1), respectively. The
Yield attributes viz. pod length (cm), number improvement in yield and yield attributing
of pods plant-1, number of grains pod-1 and character might be attributed to the fact that
test weight (Table-3 and Fig. 3) differs combined and balanced application of
significantly under the treatment T9 (RDF + Rhizobium inoculation increases
Rhizobium seed inoculation + Rhizobium photosynthetic activity of leaves,
inoculation with Vermicompost) and sole translocation of photosynthates from source to
application of Rhizobium inoculation. The sink with consequent improvement, nutrients
highest pod length (cm), number of pods uptake and better metabolism and increase
plant-1, number of grains pod-1 and test weight efficiency of other nutrients. The results of
(g) (6.9, 25.68, 5.95 and 35.42, respectively) our study are in line with the findings of
were found in T9 (RDF + Rhizobium seed Dudeja and Duhan (2005), Sahu and Singh
inoculation + Rhizobium inoculation with (2009), Kachhave et al., (2009) and Sardar et
Vermicompost) while, lowest in control (T1). al., (2016).
In the process of tissue differentiation from
somatic to reproductive, meristmatic activity From the above study it is concluded that
and development of floral primordial might application of Rhizobium treated seed with
have increased with Rhizobium inoculation Rhizobium treated vermicompost and
methods resulting in more flowers and pods recommended fertilizers was found superior
and ultimately the higher grain yield. The than rest of the treatments in terms of growth,
result is supported by Singh and Pareek nodulation and yield parameters of black
(2003), Sripriya et al., (2005), Kumar and gram. Its application resulted in higher values
Elamathi (2007) and Ghosh and Joseph for all the different aspects studied above.
(2008). Thus for obtaining higher productivity of
black gram and sustainable soil health the
Yield Rhizobium treated seed should be sown with
Rhizobium treated vermicompost and
The maximum grain yield of 11.85 q ha-1 recommended fertilizers.
(Table-4 and Fig. 4), statistically at par with
T8 (RDF + Rhizobium seed inoculation + References
Rhizobium soil treatment) and significantly
higher than remaining treatments was found Anonymous. (2019) Agricultural Statistics
in T9 (RDF + Rhizobium seed inoculation + Division, Directorate of Economics and
Rhizobium inoculation with Vermicompost), Statistics.
which were higher over control by 90.82 and Bhuiyan, M.M.H., Rahman, M.M., Afroze, F.,
84.05% in T8 and T9, respectively. Similar Sutradhar, G.N.C. and Bhuiyan, M.S.I.
result was found for harvest index with values (2008) Effect of phosphorus,
highest in T9 i.e. 26.70 % and lowest in molybdenum and Rhizobium
control i.e. 21.29 %. The highest straw and inoculation on growth and nodulation
biological yield (32.53 and 44.38 q ha-1) were of mungbean. J. Soil. Nature 2(2): 25-
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inoculation + Rhizobium inoculation with Dhakal, Y., Meena, R.S. and Kumar, S.
Vermicompost) followed by 32.41 q ha-1 and (2016) Effect of INM on nodulation,

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How to cite this article:

Shubha Tripathi, Satendra Kumar, Mukesh Kumar, Ashok Kumar, B. P. Dhyani and Yogesh
Kumar. 2021. Effect of Rhizobium Inoculation Methods on Growth, Nodulation and Yield of
Black Gram (Vigna mungo L.). Int.J.Curr.Microbiol.App.Sci. 10(01): 1588-1598.
doi: https://doi.org/10.20546/ijcmas.2021.1001.185

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