The productivity of our livestock often remains low due to inadequate and nutritionally unbalanced supply of feed and fodder. Increasing human population and reduction in land availability for cultivation are the two threats for agricultural sustainability (
Shahbaz and Ashraf, 2013). Forage is the cheapest source of animal feed which need to be properly managed, fertilized and harvested at proper stage of growth. In view of these facts, there is an urgent need of increasing good quality forage supply by adopting improved agronomic techniques such as liquid microbial inoculants. Integrated nutrient management is propounded as a promising strategy for addressing such challenges. Microbial inoculants being an important component of INM are eco-friendly and economical sources of nutrient. The use of microbial inoculants in agriculture has substantially increased during the past two decades. Thereupon, the objective of the current study was to evaluate the potential of liquid microbial inoculants to improve growth and yield of cowpea under field conditions. The results obtained from the present investigation as well as pertinent discussion have been summarized under following heads.
Growth characteristics
The data on growth parameters
viz. vine length (cm), leaf length (cm), leaf breadth (cm), stem girth (cm), number of branches per plant, number of leaves per branch and leaf: stem ratio were recorded at harvest.
Vine length (cm)
Vine length is a reliable growth index of the plant particularly fodder crops, which represents the infrastructure build-up over a period of time. It can also be considered as important factor to judge the vigour of plant.
Liquid microbial inoculants showed statistically significant effect on vine length of shoot. Maximum vine length of shoot was observed in treatment T
10 (210.47 and 205.33 cm) followed by T
5 (200.41 and 202.22 cm), T
11 (198.96 and 200.27 cm), T
6 (196.31 and 190.62 cm), T
8 (194.50 and 185.75 cm), T
7 (189.12 and 187.75 cm), T
9 (187.14 and 184.60 cm), T
3 (184.46 and 186.17 cm), T
2 (183.56 and 185.26 cm), T
4 (182.73 and 179.33 cm) and T
1 (181.05 and 176.90 cm) at Ludhiana and Bathinda respectively (Table 1 and 2). Improvement in vine length might be due to more development and branching of roots due to secretion of plant growth promoting hormones by
Burkholderia seminalis, Burkholderia sp
. and
Bradyrhizobium sp. This in turn must have helped in improved uptake of nutrients and enhanced plant vigour. These findings were similar with those of
El-Waraky and Kasem, (2007) who pointed out that inoculation of cowpea seeds cv. Kafer El-Sheikh-1 with biofertilizer (Halex-2: a biofertilizer containing a mixture of non-symbiotic Nitrogen-fixing bacteria of the genera
Azospirillum,
Azotobacter and
Klebsiella) significantly increased plant height, number of leaves and branches. Similarly,
Singh et al., (2006) reported that inoculation of rhizobium to seeds @ 0.5 kg/ha, VAM (vesicular arbuscular mycorrhiza) inoculated to soil @ 5 kg/ ha, or
Rhizobium + VAM) resulted in the highest number of leaves, vine length and number of nodules per plant of cowpea cv. Narendra Lobia-2.
Leaf length (cm)
Leaf length of plant indicates the biomass of plant. Productivity and photosynthetic activity is also correlated with the leaf area. The maximum leaf length was exhibited by treatment T
10: 75% of RDF +
Burkholderia sp
. +
Burkholderia seminalis (11.77 and 11.53 cm) and minimum at T
1: RDF (10.48 and 10.19 cm). All treatments with liquid inoculants showed significant improvement in leaf length at both locations (Table 1 and 2). The possible reason for improved leaf length might be due to increase in phosphorus availability due to phosphate solubilizing activity of cultures that gives rapid and vigorous start to plant and thus, enhanced growth parameters. Our findings were similar with
Fernandes and Bhalerao (2015) who reported that the plants treated with biofertilizer (
Rhizobium japonicum) showed significant improvement in the growth parameters like the length of leaves, breadth of leaves, shoot length and root length.
Azospirillum inoculation also significantly increased the growth in terms of length and breadth of leaf and fresh and dry weight/ plant of rice plant as reported by
Hossain et al., (2015).
Leaf breadth (cm)
Leaf breadth serves as a reliable criterion to assess leaf area which in turn relates to green fodder yield. Liquid microbial inoculants significantly improved leaf breadth at both the locations. Maximum leaf breadth (8.97 and 9.14 cm) was observed with treatment T
10 which was at par with treatment on T
5 (8.94 and 9.09cm) in both the locations (Ludhiana and Bathinda) respectively. Minimum leaf breadth was recorded with treatment T
1 (8.52 and 8.03 cm) across Ludhiana and Bathinda respectively (Table 1 and 2). This increase in leaf breadth might be due to application of liquid microbial inoculants which might have enhanced the availability of nitrogen and phosphorus by biological nitrogen fixing and phosphate solubilizing ability of
Burkholderia seminalis, Burkholderia sp. and
Bradyrhizobium sp. in addition to secretion of plant growth promoting hormones like IAA, GA and Cytokinins. These findings were similar with those of
Sharma et al., (2018) who reported that the combined application of 100 per cent RDF + biofertilizers + vermicompost resulted in maximum leaf length (61.80 cm), leaf width (22.53 cm), curd yield per hectare (35.97 tonnes/ha) and dry matter content (9.88%) in broccoli [
Brassica oleracea (L.)
var. italica]
.
Stem girth (cm)
Stem girth is an important vigour contributing component which corresponds to structural build-up of plant. The maximum stem girth was shown by the treatment T
10: 75% of RDF +
Burkholderia sp. +
Burkholderia seminalis (2.13 and 2.27 cm) and minimum values in T
1: RDF (2.00 and 1.96 cm) at Ludhiana and Bathinda respectively (Table 1 and 2). These results were supported by
Nadeem et al., (2016) who reported that combined inoculation of seed with
Rhizobium + phosphate solubilizing bacteria along with 40 kg phosphorus/ ha (P
2) significantly increased the stem girth (1.84 cm) in cowpea over rest of treatment combination. Thus, increase in stem girth with treatments of liquid microbial inoculants might be due to increase in availability of nitrogen which in turn involves cell division, enlargement, elongation and differentiation.
Number of leaves/branch
Productivity of crop depends on the process of photosynthesis, which in turn depends on number of leaves in plants. Liquid microbial inoculants significantly affected the number of leaves per branch. However, maximum number of leaves per branch was observed in the plants supplemented with T
10: 75% of RDF +
Burkholderia sp
. +
Burkholderia seminalis (42.38 and 41.99) followed by T
5: RDF +
Burkholderia sp. +
Burkholderia seminalis (41.78 and 41.63) at both the locations respectively (Table 1 and 2). The increase in number of leaves per branch might be due to more availability of nutrients such as nitrogen and phosphorus due to treatments of cowpea seeds at the time of sowing with liquid microbial inoculants. These results are in conformity with the findings of
Nkaa et al., (2014) who reported that phosphorus fertilizer significantly enhanced growth and yield characters of cowpea varieties such as number of leaves and number of branches in all the weeks of measurement. The number of leaves also increased in combined inoculation of liquid biofertilizer treatments such as
Rhizobium +
Azospirillum +
Azotobacter (T
7) at 60
th day (27.6±3.2) followed by other treatments as reported by
Uma Maheswari and Elakkiya, (2014).
Number of branches/plant
Number of branches/ plant is one of the quantitative character determining the total fodder yield. Liquid microbial inoculants non-significantly affected the number of branches/ plant. However, numerically maximum number of branches/plant was observed in treatment T
10: 75% of RDF +
Burkholderia sp. +
Burkholderia seminalis (4.27 and 4.16) followed by T
5: RDF +
Burkholderia sp. +
Burkholderia seminalis (4.22 and 4.07) at both the locations respectively. Minimum number of branches /plant was noted in the T
1: RDF (3.63 and 3.67) at Ludhiana and Bathinda respectively (Table 1 and 2). These observations are in conformity with the works of
Abdel-Hady, (2009) who demonstrated that fertilizing cowpea cv. Kaha with combination of FYM and Biofertilizer (Phosphorine + Rhizobacterien) and 1/2 NPK (recommended doses) produced the highest number of branches/plant, highest pod length, highest number of seeds /pod, highest pod filling and protein contents when compared to the control untreated plants.
Leaf stem ratio
The leaf stem ratio is an important yield contributing parameter and has a direct effect on final green and dry fodder yield of cowpea. However, maximum leaf stem ratio was observed with treatment T
10: 75% of RDF +
Burkholderia sp. +
Burkholderia seminalis (0.493 and 0.489) followed by treatments T
5: RDF +
Burkholderia sp. +
Burkholderia seminalis (0.487 and 0.480) in both the locations (Table 1 and 2). The improvement in leaf stem ratio might be attributed to the rapid expansion of dark green foliage due to increase nitrogen availability by
Burkholderia seminalis, Burkholderia sp. and
Bradyrhizobium sp. due to biological nitrogen fixation. Similar results were studied by
Verma et al., (2017) who have reported that growth parameters like leaf stem ratio, plant height, number of leaves, fresh weight of leaf, fresh weight of stem, fresh weight of plant, LAI, dry matter accumulation on leaf, stem and plant and CGR improved significantly with 100% RDN @ 60 kg ha
-1 + ST with
Azotobacter +
Azospirillum as compare to other treatments.
Green fodder yield (q/ha)
Green fodder yield represents the total biomass of plant organs and effective absorption of nutrient elements. Application of liquid microbial inoculants of
Burkholderia seminalis, Burkholderia sp. and
Bradyrhizobium sp
. have no significant effect on biological yield of forage cowpea. However, the maximum green fodder yield was obtained with treatment T
10: (285.71 and 267.85 q/ha) followed by T
5 :(RDF+
Burkholderia sp. +
Burkholderia seminalis) (280.95 and 264.28 q/ha), T
8: 75% of RDF +
Burkholderia seminalis (277.81 and 263.09 q/ha), T
11: 75% of RDF+
Burkholderia sp
. +
Bradyrhizobium sp., (277.19 and 263.90 q/ha), T
7: 75% of RDF +
Burkholderia sp. (276.19 and 255.95 q/ha), T
6: RDF +
Burkholderia sp.+
Bradyrhizobium sp. (276.38 and 263.09 q/ha), T
9: 75% of RDF +
Bradyrhizobium sp. (273.81 and 257.14 q/ha), T
3: RDF +
Burkholderia seminalis (269.04 and 254.76 q/ha), T
2: RDF +
Burkholderia sp. (266.66 and 253.57 q/ha), T
4: RDF +
Bradyrhizobium sp (263.09 and 251.19 q/ha) and minimum green fodder yield was observed with T
1: RDF (252.38 and 238.09 q/ha) at Ludhiana and Bathinda, respectively (Table 3).
Similar results were reported by
Ramanjaneyulu et al., (2010) who reported green and dry fodder yield of sorghum to be highest with half RDF (30 kg N+15 kg P
2 O
5 ha
-1) and biofertilizers (
Azotobacter + phosphate solubilizing bacteria).
Prasad et al., (2013) also reported that phosphorus along with
Rhizobium proved to be beneficial in increasing the shoot and root biomass. Similarly,
Khan et al., (2017) reported that application of
Burkholderia sp. and
Pseudomonas aeruginosa with 50% of recommended nitrogen, phosphorus and potassium fertilizers produced equivalent or higher grain yield of rice compared to the control grown with full recommended fertilizer doses, which suggests that these strains may have the potential to be used as bioinoculants for sustainable rice production.
The increase in biological yield might be because of several factors such as release of growth promoting substances like IAA, GA and control of pathogens in addition to nitrogen fixation and phosphate solublization by liquid microbial inoculants (
Burkholderia seminalis, Burkholderia sp
. and
Bradyrhizobium sp).
Dry matter yield (q/ha)
Dry matter yield is usually determined on
dry matter basis, which is weight of plant material after the moisture within the plant material has been extracted
. Liquid microbial inoculants showed statistically significant effect on
dry matter yield. Dry matter yield attained with treatment T
10 was highest (40.13 and 38.18 q/ha) followed by T
5 (40.08 and 38.00 q/ha), T
11 (39.38 and 35.79 q/ha), T
6 (39.36 and 37.51 q/ha ), T
8 (38.58 and 37.51 q/ha),T
7 (38.09 and 35.02 q/ha), T
9 (36.94 and 34.69 q/ha), T
3 (36.54 and 34.29 q/ha),T
2 (37.07 and 32.72 q/ha) and T
4 (35.21 and 31.94 q/ha) and minimum dry matter yield was observed at T
1 (34.92 and 31.76 q/ha) at Ludhiana and Bathinda respectively (Table 3).
These results were supported by
Prasad et al., (2017) who reported that the inoculation of
Rhizobium and phosphate solubilizing bacteria along with 20 kg N/ha through vermicompost significantly increased the growth
i.e. plant dry weight, crop growth rate. The increase in dry matter yield with liquid inoculant treatments indicated the favourable response of forage cowpea to seed treatment with liquid inoculants of
Burkholderia seminalis,
Burkholderia sp. and
Bradyrhizobium sp.
Soil chemical and microbial properties
In present study application of liquid microbial inoculants treatments have non-significant effect on organic carbon, available nitrogen, available phosphorus and available potassium in comparison to control (T
1) (Fig 2). This might be due to the fact that inoculated bacterial cultures were endosymbiotic in origin and involved preferential invasion of the host tissues followed by inhabitation of the root nodules. Therefore, such bacteria can affect the host plant by exhibiting direct exchange of the PGP substances and other compounds via bacteria-plant cell membrane interfaces particularly the bacteria-secreted phytohormones alter the hormonal homeostasis of the plant and lead to higher growth of the root tissue and thus increase the surface area for the absorption and uptake of the water and nutrients from the rhizosphere. However, the impact on the rhizosphere soil remains marginally low to negligible due to the preference of these bacteria to remain closely adhered to or invade the host tissues and thus exist in low numbers in the rhizosphere soil. Further, the improvement in plant yield might be attributed to increased availability of nutrients in direct vicinity of plant’s roots.
Similar to no or negligible change in the soil nutrient contents, a little numerical improvement in the total bacterial, fungal and actinomycetes count of the rhizospheric soil was recorded (Fig 3). This might be due to the fact that applied bacterial inocula moved inside the plant tissues resulting in probably no release of the molecules in the rhizosphere for the elicitation of the native microbial population. Further, the existing diversity of the native microbial communities get buffered or balanced such that the total viable counts of microbial populations remain same. Microbes which are favourable for plants get increased and others get decreased.
Kennedy, (1999) reported that the modification in bacterial community structure caused by inoculation could be buffered by ecosystem resilience, which is driven by the level of diversity and interactions of the plant-soil-biota.
It is believed that for improving soil health by use of inoculants, recurrent inoculations with highly competitive strains over a period of several years is required. As native strains give tough competition to inoculated strains (applied in the form of inoculants). Thus, bulk inoculation of soil in addition to seed inoculation is need of hour for improving soil health as these could play significant role in decomposition of organic component and myriad of biogeochemical cycles.