Research Article Nodulation, Growth and Yield …Nodulation, Growth and Yield Response of Five Cowpea (Vigna unguiculata L. Walp) Varieties to Inoculum (Mesorhizobium loti) under Different

Nodulation, Growth and Yield Response of Five Cowpea (Vigna unguiculata L. Walp) Varieties to Inoculum (Mesorhizobium loti) under Different Watering Regimes


*1OYEWUSI, Isaac Kayode, 2AGELE, Samuel Ohikhene, 3THOMAS, Oladele Victor 1Department of Agricultural Technology, The Federal Polytechnic, PMB 5351, Ado-Ekiti, Ekiti State, Nigeria 2Department of Crop, Soil and Pest Management, Federal University of Technology, Akure, Ondo State, Nigeria 3Department of Horticultural Technology, The Federal Polytechnic, PMB 5351, Ado-Ekiti, Ekiti State, Nigeria

The experiment was carried out in the screen house of the Department of Crop, Soil & Pest Management, Federal University of Technology, Akure, Nigeria. The experimental layout was a 5 x 3 x 2 factorial combination with 3 replications given a total of 90 treatments. Seeds of five cowpea varieties namely: IT98K-205-8, Ife Brown, Oloyin Brown, IT98K-573-2-1 and IT96D-610 were sown in Plastic buckets of 7-liter capacity and were perforated at the bottom to allow for drainage and filled with top soil. Watering regimes of (500ml, 700ml and 900ml) were imposed and water was applied twice a week while cowpea plants were inoculated with 5g each of Rhizobia strain (Mesorhizobia loti) at seedling stage. Control set was maintained without inoculation. The effect of watering regimes on legume species was significant on nodulation, growth and yield characters of cowpea varieties evaluated. The results revealed marked varietal differences in plant growth, nodulation, yield and yield components. IT98K-573-2-1 and Oloyin Brown generally expressed superior performance in most measured parameters. Mesorhizobia inoculation significantly (p≤0.05) increased plant growth, nodulation, yield and yield components of cowpea. The interaction effect of variety, Mesorhizobia loti and watering regimes caused significant variations in the number of nodules, leaf area, number of seeds/pod and seed yield. The nitrogen and crude protein content in the leaf differed among the cowpea varieties evaluated. Application of mesorhizobium strain significantly increased seed yield of cowpea and caused substantial increase in nodulation and this subsequently affected the Nitrogen fixation potential of cowpea under varying soil moisture regimes.

Key words: Mesorhizobia inoculation, Watering regime, Cowpea varieties Nodulation, Seed yield INTRODUCTION Cowpea production like most other crop is limited by factors such as poor yielding varieties and nutrition, absence of the right strain of rhizobia, among other factors of production (Chiezey et al., 1990). They complement staple low-protein cereal crops as a source of protein and minerals (Gharti et al., 2014). Cowpeas are also valued for their ability to fix atmospheric nitrogen into the soil and play an important role as a rotation crop with cereals and vegetable crops (Jensen et al., 2012; Biswas and Gresshoff, 2014; Stagnari et al., 2017). They also serve as a feed crop in many farming systems and are also grown to supplement farmers’ income (Muli and Saha, 2002; Voisin et al., 2013). Cowpea has considerable adaptation to high temperatures and drought compared to other crop

species, making it suitable for cultivation in semiarid areas (Hall, 2004; Tekle, 2014). Despite its role in improving soil fertility and serving as a food security crop, little efforts are made to know the response of cowpea varieties to Mesorhizobia inoculation in Nigeria. Inoculation of legumes with rhizobia strains is a common agricultural practice intended to promote nodule development and

*Corresponding Author: OYEWUSI, Isaac Kayode; Department of Agricultural Technology, The Federal Polytechnic, PMB 5351, Ado-Ekiti, Ekiti State, Nigeria. Email: [email protected] Co-Author 2Email: [email protected] 3Email: [email protected]

International Journal of Plant Breeding and Crop Science

Vol. 7(1), pp. 677-688, June, 2020. © www.premierpublishers.org, ISSN: 2167-0449

Research Article


Oyewusi et al. 678

encourage nitrogen fixation to increase legume yield (Agele et al., 2017). Plant response to inoculation is determined by a variety of factors. The presence and quality of indigenous rhizobial populations, soil nitrogen (N) availability, soil physicochemical constraints and climatic conditions significantly influence the ability to achieve increased crop yield through inoculation (George et al., 2007). Rhizobium is a common soil bacterium, not toxic to humans, plants or animals. It is one of the most beneficial bacteria to agriculture Lindermann, (2015). Rhizobia are minor components of the soil micro flora and reach their maximum numbers in association with plant roots (Bagayoko et al., 2000). They are stimulated by carbon compounds in plant roots and are controlled by microbial competition. Ditta et al., (2018). Their ability to infect legume roots and multiply within the resulting root nodules protected from the soil environment provides a special advantage over their competitors (Ahmed et al., 2008). Furthermore, Mesorhizobia inoculation of cowpea is not a common agronomic practice among smallholder farmers. Thus, the development of new cultural practice which enhance yield and mineral nutrition of grain is imperative to achieve food and nutritional security in the country.

Water stress is one of the major causes of reduced growth, development and yield in leguminous plants and also reduces nitrogen fixation and its related traits Pandy et al., (1984. Cowpeas are very sensitive to drought which leads to reduced yield and seed quality. Water stress is a major factor affecting symbiosis and leads to decreased nodule formulation, reduced nodule size and Nitrogen fixation Graham, (2002). Leguminous plants in association with rhizobium species have the potential to fix large amount of atmospheric nitrogen which contributes to the soil nitrogen pool provided that the nitrogen fixation is not restricted by other environmental or microbial factors Kennedy & Cockings (1997, Ullah et al., 2020). A more effective and easy way of raising the N status of the soil is to exploit the ability of legumes to fix appreciable quantities of atmospheric nitrogen. (Sarfraz et al., 2019). Therefore, the specific objectives of the study were to examine the effects of watering regime and Mesorhizobia inoculation on the growth, seed yield and yield components of some cowpea varieties as well as the leaf chlorophyll concentration among the tested cowpea varieties.

MATERIALS AND METHODS

Experimental site and ambient conditions

The experiment was conducted in the Department of Crop, Soil and Pest Management, Federal University of Technology Akure, located in the rain forest area of South Western Nigeria between February and April, 2016 The experiment was carried out to examine the interplay of soil moisture regimes and rhizobium inoculation on nodulation, growth and seed yield of cowpea varieties.

Planting materials

Seeds of five cowpea varieties and Rhizobia strain were obtained from the International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria. The varieties were: IT98K-205-8, Ife Brown, Oloyin Brown, IT98K-573-2-1 and IT96D-610. Plastic buckets of 7 liter capacity were perforated at the bottom and filled with top soil. Watering regimes of (500ml, 700ml and 900ml) were imposed and water was applied twice a week while cowpea plants were inoculated with 5g each of Rhizobia strain (Mesorhizobia loti) at seedling stage. The inoculant was applied close to the crop root zone at a depth of 5 cm using the procedure of seed and soil inoculation developed by Fatima et al., (2007). A Control set up was maintained without inoculation.

Treatments and experimental design

The treatments imposed on the cowpea varieties were rhizobial strain (Mesorhizobia loti) with or without rhizobium inoculation. The 5 x 3 x 2 factorial combinations (These consisted of 5 cowpea varieties, 3 watering regimes, inoculated or not) were arranged using Completely Randomized Design (CRD) with three replications given a total of 90 treatments. Based on preliminary reports on literatures, three watering regimes were imposed namely; 900ml, 700ml and 500ml and these were maintained throughout the experiment.

Data collection

Measurement of agronomic variables commenced three weeks after planting and lasted for a period of two months. Growth indices measured were vine length (cm), number of branches, number of leaves, number of nodules, size of nodules (g), weight of nodules (g), leaf area (cm3), days to 50% flowering, number of peduncles, number of seeds per plant and seed yield (kg/ha). Leaf samples were taken from the uppermost leaves of each cowpea variety during the vegetative growth stage of cowpea from each treatment. They were then analyzed for chlorophyll concentration, nitrogen, and crude protein content. The concentrations of leaf chlorophyll were determined after extraction in acetone using spectrophotometer device. (Dey et al., 2016). Crude protein was determined by multiplying the result of the total Nitrogen obtained by 6.25. The number of leaves per plant: This was estimated by counting. The plant height: This was measured using measuring tape. The measurement was taken from the base to the apical shoot. The number of branches per plant: This was estimated by counting. Number of nodules: This was done by visual counting of nodules borne on root after terminating the experiment. Nodule weight: The nodules of each plant were removed and weighed using a weighing balance. Nodule diameter: This was measured with the use of Vermeer caliper Number of pods: This was done by visual counting of pods borne on each plant at harvest. Number of seeds: This was done by visual counting of seeds after removal from pod.


Int. J. Plant Breed. Crop Sci. 679

Seed weight: This was done by weighing the seeds on weighing balance. The date to 50% flowering: This was determined when the plants produces maximum flowers during the growth stage. (Allito et al., 2015, Chatterjee and Bandyopadhjay, 2017, Dew and Singh, 2014).

Determination of Selected Soil Physical and Chemical Properties

The soil was air-dried to reduce the moisture content after which it was taken to the laboratory where the soil textural class was determined as sandy-loam as defined according to FAO/USDA soil classification system. Soil pH was determined by using 1:1 water suspension and adding 10 ml distilled water to 10 g of soil. Organic carbon was determined by modified Walkley–Black wet oxidation method. Soil organic matter of the soil sample was calculated by multiplying the organic carbon by a Van Bemmelen factor of 1.724. Potassium and sodium in the soil extract were determined by flame photometry. Flame photometer readings for the standard solutions were determined and a standard curve constructed. Potassium and sodium concentrations in the soil extract were read from the standard curve. Available P was determined using the Bray P1 method. The exchangeable magnesium alone was calculated by subtracting the value obtained from the calcium + magnesium value. Extraction and Determination of Leaf Chlorophyll

The two uppermost leaves of cowpea varieties from each treatment were harvested. One gram of the fresh plant samples were cut into pieces and smashed in a mortar. The samples were put in a test tube and its chlorophyll content was repeatedly extracted with successive volume of 100 ml acetone/water (80:20 v/v) until no traces of green colour were noticed (residue became white). While adding the solvent (acetone), the test tubes containing the samples were kept boiling in hot water bath. The total volume of the extract was also recorded at the end of the extraction. Three millimeter (3 ml) of the extract was taken and the absorbance was determined with a spectrophotometer (Spectronic 20) at two wave lengths of 663nm and 645 nm that corresponds to maximum absorption of chlorophyll “a” and “b” respectively. The total chlorophyll content was calculated as follows:

Total chlorophyll content (mg/100 g tissue) = (20.2A645 + 8.02A663) (V/10 w) Where, A645 = absorbance at 645 nm wavelength; A663 = absorbance at 663 nm wavelength, V = final volume (cm3) of chlorophyll extract in 80% acetone W = fresh weight (g) of tissue extracted (Dey et al., 2016)

Determination of crude protein

Crude protein was determined by multiplying the result of the total Nitrogen obtained by 6.25.

Statistical analysis Data collected on plant and soil parameters were subjected to analysis of variance (ANOVA) and treatment means were separated using Least Significant Difference (LSD) at 5% level of probability and Duncan Multiple Range Test (DMRT). RESULTS Pre-experiment Soil Chemical Properties The physio-chemical characteristics of the experimental soil in Table 1 show the soil has pH of 6.9. Organic matter, 4.01%, Nitrogen, 0.07 g/kg, Available P, 1.73 mg/kg, K, 0.02 cmol/kg, Na 0.02 cmol/kg, Ca 1.47cmol/kg and Mg, 4.20 cmol/kg. Table 1: Pre soil chemical properties at experimental site

Chemical Properties Values Ph (water) 6.90 Nitrogen (g/kg) 0.07 Available Phosphorus (mg/kg) 1.73 Exchangeable Na(cmol/kg) 0.02 Exchangeable K (cmol/kg) 0.02 Exchangeable Ca (cmol/kg) 1.47 Exchangeable Mg (cmol/kg) 4.20 Soil Organic matter (%) 4.01

Effects of Variety on Growth, Yield and Yield Components of Cowpea The result indicates that there was no significant difference in the number of branches, number of nodules, size and length of nodules among the tested cowpea varieties. Shoot weight was highest for IT98K-573-2-1 (114 g). The result also indicates that there was a significant difference in the leaf area of the tested cowpea varieties. Leaf area was highest for Oloyin Brown (377.8 cm3). Days to 50% flowering were shortest for IT98K-205-8 (39 days). There was no significant difference in the number of peduncles, number of pods and number of seeds among the tested varieties. Highest seed yield was recorded for Oloyin Brown (13.5 kg/ha) while the percentage crude protein was highest for Ife Brown (26.9%) as indicated in Table 2. Effect of Watering Regime (WR) on Growth, Yield and Yield Components of Cowpea Varieties More vine length was produced under 500ml of water in cowpea plant compared with the 700ml and 900ml. Table 3. This is attributed to the physiological stress occasioned by the limited water supply. Plants growing under limiting water condition tend to grow taller in an effort to scramble for nutrients around the soil. Watering regimes under 900ml significantly increased vigor and seed yield. Cowpea varieties under 900ml of water recorded the


Oyewusi et al. 680

highest number of leaves and branches for all tested varieties except IT98K-205-8. Significant difference was however observed on nodule number under 900ml of water which had the highest value. There was no significant difference among the parameters for number of branches, number of nodules, length and size of nodules, number of peduncles, 100 seed weight, harvest index and chlorophyll concentration based on the watering regimes imposed. Highest seed yield was recorded for Ife Brown (9.52 kg/ha) under the 900ml of water while there was no significant difference between the result obtained for seed yield for Oloyin Brown and IT98K-573-2-1 under 900ml and 700ml of water. Effects of Rhizobial Inoculation (RI) on Growth, Yield and Yield Components of Cowpea Varieties Rhizobium inoculation enhanced number of leaves (Table 4). More leaves were found in inoculated plant compared with the non-inoculated. Rhizobium inoculation of cowpea seedlings significantly increased vigor and seed yield. Cowpea varieties inoculated with Rhizobium recorded the highest shoot biomass as compared with the non- inoculated. Most of the agronomic characters of growth and yield components were improved by rhizobium inoculation and performed better over the non-inoculated. The result shows that rhizobial inoculation did not significantly improve nodule number, nodule size and nodule length among the cowpea varieties evaluated. Interaction Effect of Variety and Watering Regime on Growth and Yield of Cowpea The interaction effect shows significant difference among all the measured parameters. There were no significant difference in size and length of nodules, days to 50% flowering, harvest index and chlorophyll concentration among the tested cowpea varieties evaluated. The result also shows that Ife Brown performed better under 900ml of water while Oloyin Brown and IT98K-573-2-1 performed significantly better under the 900ml and 700ml of water. Interaction Effect of Variety and Rhizobia Inoculation on Growth and Yield of Cowpea Rhizobium inoculation significantly increased vigor and seed yield of cowpea. Higher number of leaves shoot biomass, leaf area and seed yield were obtained from rhizobial inoculation (Table 6). The result shows that rhizobial inoculation did not significantly improve number of nodules, size of nodules and length of nodules among cowpea varieties evaluated. There was no significant difference on rhizobia inoculation for vine length on Ife Brown, IT98K-573-2-1 and IT98K-610 while the application of rhizobia inoculation did not have any positive effect on size of nodules, length of nodules, days to 50% flowering, 100 seed weight, harvex index and chlorophyll concentration for all the tested cowpea varieties. Conversely, the number of leaves and branches, the

number of peduncles, number of pods, seed yield and crude protein levels were significantly affected by rhizobial inoculation. The highest seed yield was recorded for IT98K-573-2-1 under rhizobial inoculation (23.9 kg/ha). Interaction Effects of Variety, Rhizobial Inoculation and Watering Regime on Growth and Yield of Cowpea The interaction effect shows that significant differences were obtained for vine length, number of nodules, number of leaves and branches, leaf area, number of pods and peduncles, number of seed and seed yield while there were no significant difference for size and length of nodules, days to 50% flowering, 100 seed weight, and chlorophyll concentration in the cowpea varieties evaluated. The highest seed yield was recorded for Oloyin Brown (13.5 kg/ha) Table 7.

DISCUSSION

Effects of Watering Regimes on Growth and Yield Parameters of Cowpea

The results of this study showed that the measured growth variables differed among cowpea varieties in response to the watering regimes imposed. The enhancement of vines, number of leaves, and seed yield by heavier watering (may be attributed to higher moisture content in the crop root zone). This observation is consistent with reports of Henson et al., (2007), Majumder (2010) and Agele et al., (2012), which stated that when adequate water is available, plant cells remain turgid and plants retain their form, structure and function. In addition, Khalil and El- Noemani (2012) and Bahreininejad et al., (2013) stated that, water stress reduces plant growth through inhibition of various physiological and biochemical processes such as photosynthesis, respiration, translocation, ion uptake, carbohydrates, nutrient metabolism and hormones. This is also supported by Ullah et al., (2020) who stated that Algal biochar had a pronounced effect on Rhizobacteria which promoted growth and yield of maize under limiting water conditions. The cowpea varieties that were watered heavily with 900ml produced higher vine length, number of leaves, shoot weights and gave higher seed yields when compared with the 700ml and 500ml of water. This result further confirms how essential water is to growth and development of cowpea, a crop sensitive to water deficit (Henson et al., 2007, Majumder, 2010, Agele et al., 2012).

Effects of rhizobial inoculation on growth characters of cowpea

The cowpea varieties responded differently to rhizobial inoculation. The rhizobial inoculated plants had enhanced growth compared with the non – inoculated. These results agree with the findings of Sakthivel and Soremi, (1987), Ashrafuzzaman et al., (2009), and Isahak et al., (2012) who stated that bio-fertilizers significantly improved crop growth by increasing fixation (nitrogen) and uptake (P) of


Int. J. Plant Breed. Crop Sci. 681

soil nutrients such as nitrogen and phosphorus which influenced plant growth and significantly enhanced plant height and photosynthetic rate. Similarly, Chi et al., (2005) obtained up to 23-63% increases in plant height of inoculated rice over the non – inoculated. The authors argued that indole acetic acid (IAA) and gibberellin production were the key mechanisms for the observed growth improvement. Furthermore, the results of this study also agrees with the findings of Dar and Bali, (2007) who stated that application of bio fertilizer improved leaf development in crops. With respect to plant biomass, cowpea plants inoculated exhibited significant performance over the non- inoculated. The beneficial effect on host plant by rhizobium inoculation may be associated with improved plant nutrition. Furthermore, Hussain et al., (2020) opined that maize production can be increased by the addition of bio-activated organic fertilizer enriched with zinc solibilizing bacteria. Accordingly, these authors Ditta et al., (2018 & 2015), Ditta & Khalid, (2016), who worked on rock phosphate opined that enriched organic fertilizer with phosphate solibilizing micro-organisms improved nodulation, growth and yield of legume crops. Their combined work was supported by Zeb et al., (2018) who stressed that bacteria improved yield and Zn-fortification in flooded rice. Findings from this study showed that rhizobia inoculation did not significantly improve nodule length and size among cowpea varieties. In this study, plant nodulation was positively correlated with biomass production. Furthermore, Rapela and Saxena, (1987) stated that, the nodules of annual plants tend to die at flowering and seed set presumably because at this time, flowers and developing seeds are the major sinks of the resources thus causing nodule disintegration. Poor inoculants viability, inadequate soil mineral nitrogen, incompatibility of the inoculants strain with specific cowpea variety or the presence of highly competitive native rhizobia that restricted occupancy of the nodules by the inoculants strains are factors which affected legume response to rhizobium inoculation (George et al.,2007). In his work on rhizobacteria, Sarfraz et al., (2019) opined that growth- promoting Rhizobacteria can enhance carbon sequestration and boost crop growth. This accounted for the increased growth in cowpea varieties under rhizobial inoculation. Interaction between Variety, Watering Regime and Rhizobium Inoculation on the Performance of Cowpea The interaction effect in Table 5 shows that the performance of cowpea was influenced by watering regime and rhizobial inoculation. Seedling species that were subjected to 900ml and 700ml of water under rhizobium inoculation had enhanced growth. These findings supported the observations of Read and Boyd (1986) and Shinkafi (2000) who reported that Rhizobium inoculation increased soil water extraction and root development. Ali et al (2004), stressed that leaf emergence rate is the most sensitive parameter to drought

stress. Graham (2002) stated that mild water stress and rhizobium inoculation enhanced growth than the non- inoculated. This was consistent with the work of Osunubi and Mulongong, (1992) who reported that rhizobial inoculation enhanced leaf development. Some of the measured growth parameters of cowpea seedlings in this study were promoted by rhizobium inoculation. The highest number of pods/plant, number of seeds per pod and number of nodule was obtained from inoculated species which was significantly higher than the non- inoculated crops. This result is in agreement with Shu-Jie et al., (2007) and Agele et al (2017) who reported that the number of pods per plant, number of seeds per pod, and seed yield were significantly increased by rhizobia inoculation .This may be attributed to the symbiotic relationship between rhizobia and crop root zone (bacteria and root system of legumes). The nodule contained in roots fix atmospheric nitrogen into the roots which improves crop yield .This is in agreement with Dashti et al., (1995), Ditta et al., (2018 & 2015), Ditta &Khalid, (2016) who reported that phosphorus and rhizobium inoculation has a pronounced effect on yield and that number of pods per plant, number of seeds per pod and seed yield were significantly increased by different levels of rhizobia inoculation. In this study, there were significant (P < 0.05) interactions of rhizobial inoculation and watering regime on growth characteristics which includes leaf area, number of nodules and shoot weight on yield of inoculated cowpea. In addition, Hamidou et al., (2007) stated that water stress reduces plant growth through inhibition of various physiological and biochemical processes, such as photosynthesis, respiration, translocation, ion uptake, carbohydrates, nutrient metabolism, and hormones. This is also supported by (Sarfraz et al., (2019). The results further showed that watering regime significantly affected nodule number in all tested varieties. Soil moisture deficit stress conditions promote early flowering and maturity in crops. The reduction in leaf area and biomass accumulation, pod and seed yields were obtained under 500ml water applications. The results of this study confirmed that cowpea seedlings cannot withstand soil moisture deficit stress as was obtained for seedlings that were watered at 500ml. (Kulac et al., 2012). The 500 ml watering was characterized by high intensity of soil and air moisture deficits. These conditions have implications for survival and establishment of seedlings. Plants exposed to soil water deficit, exhibited a number of physiological responses in an effort to survive stress. These include closing of stomata and arresting cellular growth (Kulac et al., 2012). If water stress is not alleviated, plants will close stomata and shut down photosynthesis, carbon assimilation, and normal metabolism (Kulac et al., 2012). These responses means that plants growing under water stress will end up smaller and poorer in vigor. Low soil moisture stress reduces leaf area, number of leaves and branches in cowpea varieties as reported by (Sarfraz et al., (2019).


Oyewusi et al. 682 Table 2: Effects of Variety on Growth, Yield and Yield Components of Cowpea

Varieties Vine length (cm)

(6WAP

Number of

leaves (6WAP)

Number of

branches (6WAP)

Number of

Nodules

Size of nodules

(g)

Length of

nodules (cm)

Shoot wt (g)

Leaf area

(cm3)

Days to

50% Flo

No of ped

No of

pods/ plant

No of

seeds/ pod

100 Seed

Wt (g)

Seed Yield (kg/ha)

Harvex Index

Chlorophyll Conc.

(Mg/100g)

Crude protein

(%)

IT98K- 205-8

37.1d 36.1d 5.2a 6.6a 0.8a 0.6a 41.0b 293.9b 39ab 9a 7a 5a 15.3a 5.2b 0.10a 0.50a 22.9b

Ife Brown 41.4c 52.9b 5.4a 4.2a 1.1a 0.9a 14.3c 273.1b 41ab 9a 6a 6a 15.2a 7.3b 0.50a 0.40a 26.9a

Oloyin Brown

56.5a 42.7c 6.3a 5.6a 1.0a 1.1a 49.4b 377.8a 51a 8a 9a 7a 17.4a 13.5a 0.20a 0.70a 24.5a

IT98K- 573-2-1

51.9b 53.8b 5.7a 7.7a 0.8a 0.6a 114a 293.4b 43ab 10a 8a 9a 14.6a 11.2a 0.10a 0.50a 20.8b

IT96D- 610

59.4a 67.6a 4.5a 7.4a 0.9a 0.9a 106a 344.3a 43ab 7a 6a 6a 12.4ab 4.9b 0.04a 0.60a 21.4b

Means along the column bearing same superscript are not significantly different DMRT (p<0.05)(WAP) Weeks after planting. (Days to 50% flo) - Days to 50% flowering,(No of Ped) - Number of peduncles. (Chlorophyll conc) - Chlorophyll concentration

Table 3: Effect of Watering Regime (WR) on Growth, Yield and Yield Components of Cowpea Varieties

Varieties WR Vine Length (6WAP)

(cm)

Number of

leaves (6WAP)

Number of

branches (6WAP)

Number of

nodules

Size of nodules

(g)

Length of

nodules (cm)

Shoot wt (g)

Leaf area

(cm3)

Days to

50% Flo

No of ped

No of

pods/ plant

No of

seeds/ pod

100 Seed

Wt (g)

Seed Yield (kg/ha)

Harvex Index

Chlorophyll Concen- tration

(Mg/100g)

Crude Protein

(%)

IT98K- 205-8 900 32.9bc 46.7b 5.5a 9.7a 1.2a 0.8a 49.8b 313.4ab 39ab 11a 9a 5a 16.2a 6.79ab 0.133a 0.977a 26.25ab

700 31.0bc 53.0ab 5.3a 7.0a 0.8a 0.6a 47.0b 346.7ab 40ab 10a 7a 7a 16.0a 7.50ab 0.177a 0.150a 22.56ab

500 47.5b 50.7ab 4.9a 3.0ab 0.4a 0.4a 26.2cd 221.8b 37ab 7a 4ab 4ab 13.8a 2.04ab 0.097a 0.383a 19.75b

Ife Brown 900 43.0b 68.0a 6.7a 6.0a 1.3a 1.2a 19.8cd 495.6a 40ab 8a 7a 8a 17.0a 9.52a 0.675a 0.810a 28.27ab

700 39.7bc 50.7ab 4.5a 4.7ab 1.1a 0.8a 11.6cd 325.7b 43ab 10a 6ab 5a 15.8a 4.49ab 0.413a 0.317a 39.99a

500 40.6b 40.0b 4.9a 2.0ab 1.0a 0.6a 11.5cd 297.9b 41ab 8a 6ab 6a 12.7a 4.13ab 0.376a 0.120a 12.54c

Oloyin Brown 900 48.5b 54.0ab 6.5a 8.7a 1.3a 1.4a 54.0b 399.9ab 55a 8a 6ab 7a 16.7a 14.2a 0.077a 0.970a 28.62ab

700 51.1ab 41.5b 6.0a 5.0a 1.0a 1.3a 50.2b 415.4a 50a 9a 9a 7a 17.7a 11.1a 0.263a 0.790a 20.49ab

500 69.8a 32.7bc 6.5a 3.0ab 0.8a 0.6a 44.0b 318.0b 48ab 7a 12a 6a 17.9a 6.98ab 0.178a 0.197a 24.45ab

IT98K- 573-2-1 900 37.8bc 61.0ab 6.0a 9.7a 1.2a 0.8a 96.3ab 382.5a 46ab 11a 7a 10a 15.8a 12.1a 0.154a 0.717a 23.78ab

700 48.7b 51.7ab 5.5a 8.7a 0.8a 0.5a 145.0a 233.4b 43ab 10a 10a 8a 14.8a 11.4a 0.043a 0.677a 19.12b

500 69.2a 48.7b 5.7a 4.7ab 0.5a 0.6a 102ab 264.3b 40ab 8a 6ab 9a 13.1a 6.84ab 0.072a 0.180a 19.37b

IT96D- 610 900 51.5ab 77.7a 5.5a 8.7a 1.2a 1.2a 96.2ab 447.6a 46ab 9a 6ab 7a 13.0a 5.48ab 0.070a 0.657a 21.83ab

700 59.8ab 68.0a 4.3a 8.7a 1.2a 0.8a 117ab 322.8ab 44ab 7a 6ab 4ab 12.1a 4.45ab 0.053a 0.767a 19.56b

500 67.0a 57.0ab 3.7b 4.7ab 0.5a 0.6a 106ab 262.5b 39ab 4ab 6ab 7a 12.1a 3.8ab 0.017a 0.277a 22.71ab

Means along the column bearing same superscript are not significantly different DMRT (p<0.05)* significant, ns not significant at (p<0.05) (WR) -Watering regime (WAP) - Weeks after planting (No of Ped) - No of peduncles. (Days to 50% flo) - Days to 50% flowering


Int. J. Plant. Breed. Crop Sci. 683 Table 4: Effects of Rhizobial Inoculation (RI) on Growth, Yield and Yield Components of Cowpea Varieties

Variety RI Vine length (6WAP)

(cm)

Number of leaves

(WAP)

Number of branches

(WAP)

Number of nodules

Size of nodules

(g)

Length of nodules

(cm)

Shoot weight

(g)

Leaf area

(cm3)

Days to 50%

flo

No of

Ped

No of pods/ plant

No of Seeds/

Pod

100 Seed

Weight (g)

Seed Yield

(kg/ha)

Harvex Index

Chloro Conc

(Mg/100g)

Crude Protein

(%)

IT98K-205-8 + 37.8a 57.0a 5.0a 6.7a 1.5a 1.5a 30.6a 258.3b 42a 9b 10a 11a 15.8a 19.2a 0.30a 0.70a 35.9a

_ 32.9b 46.7b 5.5a 9.7a 1.2a 0.8a 49.8b 313.4a 39b 11a 9a 5b 16.2a 6.5b 0.13a 0.98a 26.3b

Ife Brown + 39.1a 86.8a 6.0a 6.0a 1.5a 1.8a 36.3a 442.0a 45a 10a 10a 9a 18.9a 17.7a 0.49a 0.94a 42.4a

_ 43.0ab 68.0b 6.7a 6.0a 1.3a 1.2a 19.8b 495.6a 40b 8a 7a 8a 17.0a 13.3b 0.68a 0.81a 28.3b

Oloyin Brown + 51.0a 78.0a 6.5a 7.7a 2.0a 2.5a 61.15a 496.6a 55a 12a 14a 10a 17.8a 10.1b 0.14a 0.66a 26.2a

_ 48.5b 54.0b 6.5a 8.7a 1.3a 1.4a 54.0b 399.9b 55a 8b 6b 7a 16.7a 14.2a 0.07a 0.97a 28.6a

IT98K-573-2-1 + 36.2a 50.7b 7.0a 10.7a 1.6a 1.4a 179.7a 222.9b 46a 13a 13a 12a 19.5a 23.9a 0.13a 0.85a 38.9a

_ 37.8a 61.0a 6.0a 9.7a 1.2a 0.8a 96.3b 382.5a 46a 11b 7b 10a 15.8a 11.8b 0.154a 0.72a 23.8b

IT96D-610 + 53.0a 80.0a 6.5a 6.7a 1.6a 1.5a 177.8a 460.5a 46a 8a 8a 8a 15.4a 9.4a 0.05a 0.65a 41.2a

_ 51.5a 77.7a 5.5a 8.7a 1.2a 1.2a 96.2b 447.6b 46a 9a 6a 7a 13.0a 6.5b 0.07a 0.66a 21.8b

Means along the column bearing same superscript are not significantly different DMRT (p<0.05) (WAP) - Weeks after planting, (RI) - Rhizobium inoculation.(Days to 50% flow) - Days to 50% flowering. (No of ped)- No of peduncles. (Chloro Conc) - Chlorophyll concentration

Table 5: Interaction Effects of Variety and Watering Regime on Growth, Yield and Yield Components on Cowpea Performance

Varieties WR Vine Length (cm)

(6WAP)

Number of leaves (6WAP)

Number of branches (6WAP)

Number of nodules

Size of nodules

(g)

Length of nodules

(cm)

Shoot weight

(g)

Leaf area

(cm3)

Days to 5o% flo

No of

ped

No of pods/ plant

No of Seeds/

pod

100 Seed

Weight (g)

Seed Yield (kg/ha

HI Chloro Conc

(Mg/100g)

Crude protein

(%)

IT98K-205-8 900 32.9bc 46.7b 5.5 9.7a 1.2a 0.8a 49.8b 313.4ab 39ab 11a 9a 5a 16.2a 6.79ab 0.13a 0.97a 26.2ab

700 31.0bc 53.0ab 5.3a 7.0a 0.8a 0.6a 47.0b 346.7ab 40ab 10a 7a 7a 16.0a 7.50ab 0.17a 0.15a 22.5ab

500 47.5b 50.7ab 4.9a 3.0ab 0.4a 0.4a 26.2cd 221.8b 37ab 7a 4ab 4ab 13.8a 2.04ab 0.09a 0.38a 19.7b

Ife Brown 900 43.0b 68.0a 6.7a 6.0a 1.3a 1.2a 19.8cd 495.6a 40ab 8a 7a 8a 17.0a 9.52a 0.67a 0.81a 28.2ab

700 39.7bc 50.7ab 4.5a 4.7ab 1.1a 0.8a 11.6cd 325.7b 43ab 10a 6ab 5a 15.8a 4.49ab 0.41a 0.31a 39.9a

500 40.6b 40.0b 4.9a 2.0ab 1.0a 0.6a 11.5cd 297.9b 41ab 8a 6ab 6a 12.7a 4.13ab 0.37a 0.12a 12.5c

Oloyin Brown 900 48.5b 54.0ab 6.5a 8.7a 1.3a 1.4a 54.0b 399.9ab 55a 8a 6ab 7a 16.7a 14.2a 0.07a 0.97a 28.6ab

700 51.1ab 41.5b 6.0a 5.0a 1.0a 1.3a 50.2b 415.4a 50a 9a 9a 7a 17.7a 11.1a 0.26a 0.79a 20.4ab

500 69.8a 32.7bc 6.5a 3.0ab 0.8a 0.6a 44.0b 318.0b 48ab 7a 12a 6a 17.9a 6.89ab 0.17a 0.19a 24.45ab

IT98K-573-2-1 900 37.8bc 61.0ab 6.0a 9.7a 1.2a 0.8a 96.3ab 382.5a 46ab 11a 7a 10a 15.8a 12.1a 0.15a 0.71a 23.7ab

700 48.7b 51.7ab 5.5a 8.7a 0.8a 0.5a 145a 233.4b 43ab 10a 10a 8a 14.8a 11.4a 0.04a 0.67a 19.1b

500 69.2a 48.7b 5.7a 4.7ab 0.5a 0.6a 102ab 264.3b 40ab 8a 6ab 9a 13.1a 6.84ab 0.07a 0.18a 19.3b

IT96D-610 900 700

51.5ab

59.8ab

77.7a

68.0a

5.5a

4.3a

8.7a

8.7a

1.2a

1.2a

1.2a

0.8a

96.2ab

117ab

447.6a

322.8ab

46ab

44ab 9a

7a 6ab

6ab 7a

4ab 13.0a

12.1a

5.48ab

4.45ab

0.07a

0.05a

0.65a

0.76a

21.8ab

19.5b

Variety (Var) 500 67.0a 9.30

57.0ab 11.2

3.7b 8.3

4.7ab 2.5 0.5a 0.4

0.6a 0.5

106ab

16.5 262.5b 21.4

39ab

8.5 4ab

3.0 6ab

2.2 7a

2.3 12.1a 4.5

3.8ab 0.11

0.01a 0.20

0.27a 0.30

22.7ab 3.5

W R Var x WR

12.2 *

8.20 *

1.3 *

3.1 *

0.3 ns

0.3 ns

12.7 *

36.2 *

2.3 ns

4.1 *

3.4 *

2.1 *

3.5 ns

2.40 *

0.09 ns

0.05 Ns

8.3 *

Means along the column bearing same superscript are not significantly different DMRT (p<0.05) (WR) Watering regime. (WAP) Weeks after planting (No of Ped) - Number of peduncles. (Days to 50% flo)- Days to50% flowering.(HI) - Harvex index. (Chloro conc) - Chlorophyll concentration (*) - significant (ns) - not significant ns


Oyewusi et al. 684 Table 6: Interaction Effects of Variety and Rhizobial Inoculation on Growth, Yield and Yield Components on Cowpea Performance

Varieties RI Vine length (6WAP)

(cm)

Number of

leaves (6WAP)

Number of

branches (6WAP)

Number of

nodules

Size of nodules

(g)

Length of

nodules (cm)

Shoot weight

(g)

Leaf area

(cm3)

Days to 50%

flo

No of ped

No of pods/ plant

No of Seeds/

pod

100 Seed

weight (g)

Seed yield (kg/ha)

Harvest Index

Chlro Conc

(Mg/100g)

Crude Protein

(%)

IT98K-205-8 + 37.8a 57.0a 5.0a 6.7a 1.5a 1.5a 30.6a 258.3b 42a 9b 10a 11a 15.8a 19.2a 0.30a 0.70a 35.9a

_ 32.9b 46.7b 5.5a 9.7a 1.2a 0.8a 49.8b 313.4a 39b 11a 9a 5b 16.2a 6.5b 0.13a 0.98a 26.3b

Ife Brown + 39.1ab 86.8a 6.0a 6.0a 1.5a 1.8a 36.3a 442.0a 45a 10a 10a 9a 18.9a 17.7a 0.49a 0.94a 42.4a

_ 43.0a 68.0b 6.7a 6.0a 1.3a 1.2a 19.8b 495.6a 40b 8a 7a 8a 17.0a 13.3b 0.68a 0.81a 28.3b

Oloyin Brown + 51.0a 78.0a 6.5a 7.7a 2.0a 2.5a 61.1a 496.6a 55a 12a 14a 10a 17.8a 10.0b 0.14a 0.66a 26.2a

_ 48.5b 54.0b 6.5a 8.7a 1.3a 1.4a 54.0b 399.9b 55a 8b 6b 7a 16.7a 14.2a 0.077a 0.97a 28.6a

IT98K-573-2-1 + 36.2a 50.7b 7.0a 10.7a 1.6a 1.4a 179.7a 222.9b 46a 13a 13a 12a 19.5a 23.9a 0.13a 0.85a 38.9a

_ 37.8a 61.0a 6.0a 9.7a 1.2a 0.8a 96.3b 382.5a 46a 11b 7b 10a 15.8a 11.8b 0.15a 0.72a 23.8b

IT96D-610 + 53.0a 80.0a 6.5a 6.7a 1.6a 1.5a 177.8a 460.5a 46a 8a 8a 8a 15.4a 9.4a 0.05a 0.65a 41.2a

_ 51.5a 77.7a 5.5a 8.7a 1.2a 1.2a 96.2b 447.6b 46a 9a 6a 7a 13.0a 6.5b 0.07a 0.66a 21.8b

Variety (Var) 9.30 11.2 8.3 2.5 0.4 0.5 16.5 21.4 8.5 3.0 2.2 2.3 4.5 0.11 0.20 0.30 3.5

RI 9.2 12.0 1.3 1.5 0.3 0.3 11.9 46.2 1.3 2.1 4.4 3.1 2.5 6.40 0.09 0.05 11.3

Var x RI Ns * Ns ns ns ns * * ns * * * ns * ns ns *

Means along the column bearing same superscript are not significantly different DMRT (p<0.05) RI) - Rhizobial inoculation (WAP) - Weeks after planting. (Days to 50% flo) – Days to 50% flowering. (No of Ped) - Number of peduncles. (Chloro conc) - Chlorophyll concentration (*) - Significant- (ns) - not significant at (p<0.05)


Int. J. Plant. Breed. Crop Sci. 685 Table 7: Interaction Effects of Variety, Watering Regime and Rhizobial Inoculation on Cowpea Performance

Varieties VL (6WAP)

(cm)

NOL (6WAP)

NOB (6WAP)

NON SON (g)

LON (g)

Shoot weight

(g)

Leaf area

(cm3)

Days to 50% Flow

No of ped

No of pods

No of Seeds

100 seed

weight (g)

Seed yield (kg/ha)

Harvex Index

CC (Mg/100g)

Crude protein

(%)

IT98K-205-8 Ife Brown Oloyin Brown IT98K-573-2-1 IT96D-610 Watering 900ml 700ml 500ml Rhizobial Non-inocul Variety (Var) Watering Reg Rhizobial

37.1d

41.4c

56.5a

51.9b

59.4a

42.7a

44.3a

29.9ab

43.4a

42.7a 34.1 12.2 9.2

36.1d

52.9b

42.7c

53.8b

67.6a

61.5a

52.4ab

45.8ab

66.7a

61.3ab 61.5 8.20 12.0

5.2a

5.4a

6.3a

5.7a

4.5a

6.0a

5.1a

5.1a

6.2a

6.0a 7.2 1.3 1.3

6.6a

4.2a

5.6a

7.7a

7.4a

8.6a

6.8a

4.4ab

8.6a

7.6a 8.6 3.1 1.5

0.8a

1.1a

1.0a

a

a

1.2a

1.0a

0.6a

1.6a

1.2a 1.2 0.3 0.3

0.6a

0.9a

1.1a

0.6a

0.9a

1.2a

0.8a

0.6a

1.7a

1.1a 1.1 0.3 0.3

41.0c

14.3d

49.4b

114a

106a

63.2b

74.2ab

57.9b

97.1a

63.2b 63.2 12.7 11.9

293.9b

273.1b

377.8a

293.4b

344.3a

407.8a

328.8b

272.9c

376.9b

407.8a 407.8 36.2 46.2

39ab

41ab 51a

43ab

43ab

45a 44a

41ab

47a 45a 45 2.3 1.3

9.0a

9.0a

8.0a

10.0a

7.0a

9.4a

9.2a

6.8a

10a

9.4a 9.4 4.1 2.1

7.0a

6.0a

9.0a

8.0a

6.0a

7.0a

8.0a

7.0a

11a

7ab

7.0 3.4 4.4

5.0a

6.0a

7.0a

9.0a

6.0a

7.4a

6.2a

6.4a

10a

7.4ab 8.2 2.1 3.1

15.3a

15.2a

17.4a

14.6a

12.4a

15.7a

15.3a

13.9a

17.5a

15.7a

15.7 3.50 2.50

5.2ab

7.3ab

13.5a

11.2a

4.9ab

9.6a

7.8ab

5.7b

16.1a

10.5b 10.5 2.40 6.40

0.10a

0.50a

0.20a

0.10a

0.04a

0.22a

0.19a

0.15a

0.22a

0.22a 0.22 0.09 0.09

0.50a

0.40a

0.70a

0.50a

0.60a

0.83a

0.54a

0.23a

0.76a

0.83a 0.83 0.05 0.05

22.9ab

26.9a

24.5a

20.8ab

21.4ab

25.8a

24.3a

19.8ab

36.9a

25.8b 25.8 8.30 11.3

Var x WR Var x RHI Var WR x RHI

* ns *

* * *

* ns *

* ns *

ns ns ns

ns ns ns

* * *

* * *

ns ns ns * * *

* * *

* * *

ns ns ns * * *

ns ns ns

ns ns ns * * *

Means along the column bearing same superscript are not significantly different DMRT (P0.05) (WAP) - Weeks after planting.(VL) - Vine length. (NOB) - Number of branches. (NOL) - Number of leaves. (NON) - Number of nodules. (SON) - Size of nodules. (LON) - Length of nodules. (Days to flow) - Days to 50% flowering, (No of Ped) - Number of peduncles. (CC) - Chlorophyll concentration. (WR) - Watering Régime. (Non-inocul) - Non- inoculation


Oyewusi et al. 686

CONCLUSIONS Cowpea growth and yield attributes were significantly affected by watering regimes and rhizobial inoculation. The results obtained in this study have shown that mesorhizobial inoculation can improve plant growth, nodulation and grain yield of the tested cowpea varieties grown in the low-N soils of Akure. The higher plant growth and increased nodulation in Rhizobium-inoculated plants translated into increased grain yield. Based on the findings of this study, IT98K-573-2-1 and Oloyin Brown responded better to the application of mesorhizobial loti and could be recommended for enhanced growth and yield performance of cowpea. The nitrogen and crude protein content in leaf differed among the cowpea varieties evaluated. Application of mesorhizobium strain significantly increased seed yield of cowpea and this caused substantial increase in nodulation and subsequently affected the N2 fixation potential of cowpea under varying soil moisture regimes. Based on the measured growth parameters in this study, rhizobium inoculation and application of 900ml of water promoted plant vigor and increased cowpea yield in the study area. COMPETING INTERESTS Authors have declared that no competing interests exist REFERENCES Agele, S.O, and Agbi O.M (2012): Growth and yield

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Accepted 30 May 2020 Citation: Oyewusi IK, Agele SO, Thomas, OV (2020). Nodulation, Growth and Yield Response of Five Cowpea (Vigna unguiculata L. Walp) Varieties to Inoculum (Mesorhizobium loti) under Different Watering Regimes. International Journal of Plant Breeding and Crop Science, 7(1): 677-688.

Copyright: © 2020: Oyewusi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are cited.

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Research Article Nodulation, Growth and Yield …Nodulation, Growth and Yield Response of Five Cowpea (Vigna unguiculata L. Walp) Varieties to Inoculum (Mesorhizobium loti) under Different