Effectiveness of different bio-fertilizer on growth, yield and yield attributing character of Faba bean (Vicia fabae L.)

doi:10.21203/rs.3.rs-4928559/v1

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Effectiveness of different bio-fertilizer on growth, yield and yield attributing character of Faba bean (Vicia fabae L.)

https://doi.org/10.21203/rs.3.rs-4928559/v1

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The excessive use of chemical fertilizers has led to soil depletion of essential minerals, causing microorganisms to be distracted and reducing the distribution of soil fauna and flora. The field experiment examined the impact of biofertilizer (Rhizobium and Phosphorus Solubilizing Bacteria) and blended fertilizer (NPKSZn) on the growth, yield, and yield-attributing characteristics of Faba bean. The experiment was laid out in RCBD with eight treatments. The result revealed that the application of biofertilizer significantly (P < 0.05) affected the growth and yield characteristics of faba bean. The longest grain filling period (51.42 days), tallest plant height (148.2cm), Maximum nodulation (79.58), number of branches (6.42), pod per plant (34.08), highest thousand seed weight (699.9 g), grain (2369 kg ha-1), and biological yield (3942 kg ha-1), and the lowest chocolate spot disease severity index (13.33%) was noticed in dual inoculation of Rhizobium and PSB along with recommended dose of NPKSZn in both growing seasons. The highest net benefit (48174.9 ETB per ha) with an acceptable maximum marginal rate of return (789.26%) was recorded in dual inoculation of seed with Rhizobium and PSB. Therefore inoculations of Rhizobium and PSB enhance the production and productivity and reduce the chocolate spot infestation level. Hence the seed must be inoculated with a dual culture of Rhizobium and PSB along with another package of practices for the crop.

Blended fertilizer

Biofertilizer

Chocolate spot

nodulation

Rhizobium

Faba bean (Vicia fabae L.) ranks among the most important pulse crops in Ethiopia and occupies about 30% of the total cultivated area [1]. Faba bean is used as human food as a green vegetable or dried, fresh, or canned in the Middle East, Mediterranean region, China and Ethiopia. Ethiopia is the second largest producer of faba bean in the world, next to China [2]. However, the national productivity of faba bean in the country is still very low. According to the report of the Central Statistics Agency CSA [3], the national average yield of faba bean under smallholder farmers‟ is 20.53 quintals per hectare [3]. In the Tigray region, faba bean covers an area of 9228.25 ha, and its production accounts for 151,091.02 quintals. Based on the CSA [3] data, the productivity of the faba bean in Tigray region is 16.37 quintals/hectare, which is lower than the national average [3]. Several abiotic factors contributed to low productivity of which poor soil fertility, the acidity of the soil in high rainfall areas, and the low existence of an effective indigenous rhizobia population in the area.

Dependence on chemical fertilizers and pesticides to increase crop yield has encouraged the thriving of industries that are producing life-threatening chemicals that are not only hazardous for human consumption but can also disturb the ecological balance [4]. This contributes to poor economic returns from the expenditure made on chemical fertilizers. Therefore, all efforts must be made to improve fertilizer use efficiency by crops. For this reason, seed inoculation with phosphate solubilizing bacteria (PSB), nitrogen-fixing microorganisms, and other beneficial microorganisms as a bio-fertilizer is an acceptable alternative to chemical fertilizer application.

Application of bio-fertilizers containing beneficial microbes promotes to a large extent, crop productivity and sustainability of soil and protects the environment as eco-friendly and cost-effective inputs for the farmers [5]. Biofertilizers keep the soil environment rich in all kinds of macro and micronutrients via nitrogen fixation, phosphate and potassium mineralization, the release of plant growth regulating substances, production of antibiotics, and biodegradation of organic matter in the soil [6]. Hence biofertilizers can be an important component of integrated nutrient management systems for sustaining agricultural productivity and a healthy environment [7].

Despite the existence of considerable knowledge on the effects of specific taxa of biofertilizers, a comprehensive quantitative assessment of the performance of biofertilizers with different traits such as phosphorus solubilization and N fixation applied to various crops at a global scale is missing. Therefore the objective of the present investigation was to evaluate the effect of different combinations of biofertilizers on the growth, yield, and yield components of faba bean.

2.1. Description of the experimental site

The experiment was carried out at Atsela and Mekelle University research sites. Atsela is located between 12°55'N latitude and 39°32'E longitude at 2569 meters above sea level. The site receives a mean annual rainfall of 911mm with mean minimum and maximum temperatures of 10.3 and 23.6°C, respectively. The Woreda is grouped as a cold sub-moist highland. Mekelle University is located at 13014 ’N latitude and 39032’E longitude and at 2220 meters above sea level. The average minimum and maximum air temperatures during the cropping season were 17.4˚C and 33.20˚C, respectively. The experimental area had brown soil with the textural class of silt loam and a pH of 7.51 [8].

2.2. Treatment and Experimental Design

Field experiment with bio-fertilizers viz. Rhizobium and Phosphorous solubilizing bacteria (PSB) inoculation and blended fertilizer (NPKSZn) on faba bean was carried out in two different sites (Atsela, and Mekelle University) in off and rain-fed seasons. The experiments were laid out in a Randomized Complete Block Design using eight treatments with three replications. The treatments were as follows: T1: Control T2: only Rhizobium inoculation, T3: only PSB inoculation, T4: only NPKSZn, T5: Rhizobium + PSB inoculation, T6: Rhizobium inoculation + NPSZ, T7: PSB inoculation + NPKSZnand T8: Rhizobium + PSB inoculation + NPKSZn. Rhizobium and PSB inoculation were used @ 250 g/10 kg seeds. PSB was collected from Menagesha Biotechnology Institute. The blended fertilizer contains NPKSZn with 15,31,8,7 and 2.2 levels applied at planting. Seeds were inoculated with Rhizobium and PSB before sowing and after drying in the shade, sowing was done at 40 cm row-to-row spacing using 45 kg seed/ha.

The faba bean variety Moti was sown on the 18th of January during off season and on the 20th of June during the rain-fed season. The plot size was 3 m x 2 m. Each plot and block was separated by a 0.5 m bund and 1 m path respectively. Each plot has 6 rows of 40 cm apart. Biofertilizers were also applied according to the treatments. The weeding was performed at 30 and 45 Days after emergence. Before harvesting, two outermost rows on both ends of the plots and 25 cm on both ends of each row were considered as borders.

2.3. Data Collection

Five plants were selected randomly from each plot for recording the growth parameters: plant height, number of pods per plant, number of seeds per pod, grain yield, biological yield, and harvest index at the end of physiological maturity.

Number of Nodules was counted from randomly selected five plats at flowering stage

Grain yield

The harvested crop of each plot was weighted in kg and then converted to ha.

Harvest index (HI): HI= (GY/ABG)*100

Disease Intensity

Data were taken at 6 days interval to calculate disease severity by using following formula:

$$\:\text{D}\text{i}\text{s}\text{e}\text{a}\text{s}\text{e}\:\text{i}\text{n}\text{c}\text{i}\text{d}\text{e}\text{n}\text{c}\text{e}=\frac{\text{N}\text{u}\text{m}\text{b}\text{e}\text{r}\:\text{o}\text{f}\:\text{i}\text{n}\text{f}\text{e}\text{c}\text{t}\text{e}\text{d}\:\text{p}\text{l}\text{a}\text{n}\text{t}\text{s}}{\text{T}\text{o}\text{t}\text{a}\text{l}\:\text{n}\text{u}\text{m}\text{b}\text{e}\text{r}\:\text{o}\text{f}\:\text{p}\text{l}\text{a}\text{n}\text{t}\:\text{a}\text{s}\text{s}\text{e}\text{s}\text{s}\text{e}\text{d}}\:\text{X}\:100$$

Disease severity index

The severity of the disease was examined visually on tagged plants along the plot and recorded as the percentage of plant parts (tissues) affected (percentage of blast infection on the plant).

$$\:\text{P}\text{e}\text{r}\text{c}\text{e}\text{n}\text{t}\:\text{s}\text{e}\text{v}\text{e}\text{r}\text{i}\text{t}\text{y}\:\text{i}\text{n}\text{d}\text{e}\text{x}=\frac{\text{S}\text{u}\text{m}\:\text{o}\text{f}\:\text{n}\text{u}\text{m}\text{e}\text{r}\text{i}\text{c}\text{a}\text{l}\:\text{r}\text{a}\text{t}\text{i}\text{n}\text{g}\:\text{x}100}{\text{N}\text{o}.\:\text{o}\text{f}\:\text{p}\text{l}\text{a}\text{n}\text{t}\text{s}\:\text{r}\text{a}\text{t}\text{e}\text{d}\:\text{x}\:\text{m}\text{a}\text{x}.\:\text{s}\text{c}\text{o}\text{r}\text{e}\:\text{o}\text{f}\:\text{s}\text{c}\text{a}\text{l}\text{e}}\:$$

2.4. Data analysis:

Analysis of variance was performed using the program Genstat version 20 [9]. Before the data were subjected to the statistical software, normality of the data distribution was performed using Skewness /Kurtosis tests using STATA-version 11. Accordingly all the data were normally distributed. The significance of the treatment effect was determined using F-test. When analysis of variance showed significant treatment effects, the Duncan test was applied to make comparisons among the means at p < 0.05 level of significance. The graph was done using Excel.

2.5. Partial Budget Analysis

Yield from experimental plots was adjusted downward by 15%, i.e. 10% for management difference and 5% for plot size differences, to reflect the difference between the experimental yield and the yield that farmers could expect from the same treatment. Accordingly, the mean grain yields for NPKSZn, Rhizobium, and PSB inoculation treatment combinations were subjected to a discrete partial budget analysis using the procedures outlined by Bezabeh et al. [10]. To estimate economic parameters, the variable cost of NPKSZn (ETB 24.8 kg-1), Rhizobium, and PSB inoculants (ETB 180 kg-1) were recorded at the time of planting. The price of the current faba bean grain was (ETB 26 kg-1). The price of blended fertilizer (NPKSZn) and Rhizobium and PSB inoculants were taken from Agricultural Inputs Supply Enterprise and Menagesha Biotech Industry PLC at the time of planting, respectively.

Gross field benefit = Adjusted Yield X Field Price

Net Benefit = Gross income –total variable cost [10].

$$\:MRR\:\left(\%\right)=\frac{\text{M}\text{a}\text{r}\text{i}\text{g}\text{i}\text{n}\text{a}\text{l}\:\text{i}\text{n}\text{c}\text{r}\text{e}\text{a}\text{s}\text{e}\:\text{i}\text{n}\:\text{n}\text{e}\text{t}\:\text{b}\text{e}\text{n}\text{e}\text{f}\text{i}\text{t}\text{l}}{\text{M}\text{a}\text{r}\text{i}\text{g}\text{i}\text{n}\text{a}\text{l}\:\text{i}\text{n}\text{c}\text{r}\text{e}\text{a}\text{s}\text{e}\:\text{i}\text{n}\:\text{c}\text{o}\text{s}\text{t}}\:\:X\:100$$

The average open market price (Birr kg-1) for Faba bean crop and the official price of blended fertilizer from Enderta Union were used for analysis.

3.1. Soil Physical properties of the experimental site

Pre-sowing soil sample analysis demonstrated that the soil exhibited low ratings for N and K, while that of P was medium in Atsela experimental sites.

Table 1

Soil Physical properties of the experimental site
Location	pH	%OC	Available P (ppm)	Available K (mg/kg)	%Total N	CEC (meq/100g)	% Texture			Textural class
Location	pH	%OC	Available P (ppm)	Available K (mg/kg)	%Total N	CEC (meq/100g)	Sand	Clay	Silt	Textural class
Atsela	6.6	4.58	11.4	23.2	0.07	21.6	38	36	26	Clay loam
MU	6.4	4.44	9.8	24.5	0.054	20.4	35	25	40	Silt loam
^{OC = organic carbon; P= Phosphorus; K= potassium; N= Nitrogen; CEC= Cation exchange capacity}

3.2. Effect of different biofertilizer on phonological traits of faba bean

Biofertilizer significantly affected days to flowering and maturity, number of branches, pod per plant, seed per pod, plant height, biomass and grain yield, and chocolate spot of faba bean.

Table 2

Analysis of variance for Days to emergence, flowering, and maturity) number of branches, nodule number, plant height, number of pods per plant, seed per pod, and thousand seed weight
SV	d.f.	DTF	DTM	GFP	NB	NN	NPPP	SPP	PH
Block	2	2.15	1.969	6.125	0.698	26.323	4.167	0.5417	86.26
Biof.	7	43.18^**	108.48^**	294.49^**	16.3^**	2539.7^**	425.5^**	5.15^**	1918.5^**
Loc.	1	98.18^**	4.18 ^ns	0.01 ^ns	32.67^**	213.1^**	90.09^**	4.59^**	677.34^**
Yr.	1	149.4^**	135.4^**	21.36^**	0.67 ^ns	189.8^**	1372.6^**	0.01 ^ns	5638.6^**
Biof.^*Loc.	7	0.01 ^ns	2.59 ^ns	0.000 ^ns	0.53 ^ns	4.296^ns	4.31 ^ns	0.12 ^ns	28.99 ^ns
Biof.^*Yr.	7	0.52 ^ns	1.9 ^ns	1.295 ^ns	0.14 ^ns	2.368^ns	42.81^**	0.06 ^ns	91.86^**
Loc.^*Yr.	1	0.02 ^ns	15.04^**	0.000 ^ns	0.0 ^ns	25.01^**	23.01^**	0.09 ^ns	677.34^**
Biof.^*Loc.^*Yr.	7	0.01 ^ns	1.6 ^ns	0.000^ns	0.14 ^ns	1.963^ns	3.94 ^ns	0.05 ^ns	28.99 ^ns
Residual	62	1.32	1.732	1.667	0.33	2.108	2.124	0.2836	17.70

Where Biof = Biofertilizer treatment; Loc = Location; Yr = Year; DTF = Days to flowering; DTM = Days to maturity; GFP = Grain filling period; NB = Number of Branch; NN = Number of nodules; PH = Plant height; (NPPP), Seed per pod NSPP = Number of pod per plant; ** = p≤ 0.01; * = p ≤ 0.05; ns = non−significant.

3.3. Days to Flowering, Maturity and grain filling period

Inoculation of seed with Rhizobium and PSB significantly (p < 0.05) influenced days to flowering, maturity, and grain filling period of faba bean (Table 2). The longest days to flowering (83 and 82 days) were observed at the control and application of NPKSZn respectively. The shortest (77.6 and 78 days) were obtained under dual inoculation of Rhizobium and PSB along with the application of NPKSZn and seed inoculated with Rhizobium and PSB respectively. In contrast to the days to flowering, the longest days to maturity (129 and 126 days) were recorded with dual inoculation of Rhizobium and PSB along with the application of NPKSZn and with combined inoculation of Rhizobium and PSB respectively. While the shortest number of days to maturity (120.8 and 121 days) were obtained with the application of NPKSZn and control respectively.

3.4. Number of branches and nodule number

The result indicated that the application of biofertilizer significantly affected the number of branches and nodule number per plant. Significant variation was also observed between the locations. The highest number of branches (6.42 and 5.67) was recorded in dual inoculation of seeds with Rhizobium and PSB along with the application of NPKSZn and combined inoculation of Rhizobium and PSB respectively. While the lowest number of branches was recorded in the control and application of NPKSZn. Dual inoculation of Rhizobium and PSB increased the number of branches by 21.41 and 44.64% over their sole application of Rhizobium and PSB respectively. Among the locations highest number of branches was obtained in Atsela site than Mekelle university experimental site.

Results of the present investigation revealed that the application of biofertilizer significantly increased the number of nodules (Fig. 1). Dual inoculation of Rhizobium and PSB with and without application of NPKSZn gave higher (79.58 and 79.25) number of nodules per plant respectively. In contrast, the lowest number of nodules per plant (40.75) was counted in plots treated with chemical fertilizer (NPKSZn) followed by control (Table 2). Growing location and season also significantly affected the number of nodules with which the highest number of nodules was obtained in atsela (63.94) and in the second growing season (63.85). The current result is in agreement with Desta et al. [11] who reported that both inoculation as sole and/or the combined effect of strains with zinc significantly increased nodule number plant − 1. Correspondingly, Bejandi et al. [12] reported that seed inoculation with Rhizobium cicerea produced significantly the highest number of active nodules per plant than control. In addition, Nagwa et al. [13] reported that inoculation of strain to faba bean significantly increased nodule number. Comparably, a study by Woldekiros et al. [14] disclosed that nodule number per plant increased with increasing potassium supply on of faba bean and other legume crops.

3.5. Number of pod per plant and Seeds per pod

The present result revealed that pod number per plant of faba bean was significantly affected by bio-fertilizers, growing season, and location (Table 2). In both experimental sites, a higher number of pods per plant (34.08) were obtained in dual inoculation of Rhizobium and PSB along with the application of NPKSZn followed by combined inoculation of Rhizobium and PSB (Fig. 2). However, the lowest pod numbers (14.67 and 19.17) were recorded in control and application of NPKSZn without inoculation (Table 2). Across the location maximum number of pods was recorded at Atsela than Mekelle University. Dual inoculation of Rhizobium and PSB increased pod number by 16.31 and 30.12% over their sole application of Rhizobium and PSB respectively and by 98.84 and 52.16% as compared to the control and NPKSZn respectively. A higher number of pods due to inoculation and NPKSZn application suggested that there was a better combining and symbiotic relationship between introduced Rhizobium and PSB with faba bean. This is in agreement with the findings of Abbasi et al. [15] who reported that inoculation with B. japonicum significantly, increased the number of pods per plant of soybean as compared to the control treatments.

Table 3

Main effect of bio-fertilizers, location and growing season on growth and yield related traits of faba bean
Treatment	DTF	GFP	DTM	NB	NN	PH	SPP	NPPP
Rzm + PSB + NPKSZn	77.6^e	51.42 ^a	129.4^a	6.42 ^a	79.58^a	148.2 ^a	4.5 ^a	34.08 ^a
Rzm + NPKSZn	79.8^bc	44.68^c	125.0^c	5.08^c	69.17^b	132.5^c	4.1^abc	26.67^c
RZM	79.1^bc	44.55^cd	124.2^cd	4.67 ^c	68.50^b	124.3^d	3.8^bcd	25.08^d
Rzm + PSB	78.1^de	47.90^b	126.8^b	5.67^b	79.25^a	138.4^b	4.3^ab	29.17^b
PSB + NPKSZn	78.9^cd	43.55^d	123.2^d	4.67 ^c	60.92^c	123.4^d	3.7^cd	24.17^d
PSB	79.9^b	39.85^e	121.5^e	3.92^d	56.83^d	123.3^d	3.5^d	22.42^e
NPKSZn	82.2^a	37.56^f	120.8^e	3.25^e	40.75^f	115.0^e	2.9^e	19.17^f
Control	83.1^a	37.27^f	121.1^e	3.00^e	44.58^e	108.8^f	2.6^e	14.67^g
LSD(5%)	0.937	1.054	1.074	0.47	1.185	3.433	0.435	1.189
CV(%)	1.4	3.0	1.1	12.6	2.3	3.3	14.5	6.0
Atsela	80.86	43.35	124.21	5.17	63.94	129.42	3.90	25.40
MU	78.84	43.35	123.79	4.00	60.96	124.10	3.46	23.46
Off season	78.60	43.82	122.81	4.50	61.04	102.54	3.69	20.65
Rain fed	81.10	42.88	125.19	4.67	63.85	150.98	3.67	28.21
LSD	0.469	0.527	0.537	0.235	0.592	1.717	0.217	0.595
^{RZM = (Rhizobium); PSB = (Phosphorus solubilizer Bacteria) CV = (coefficient of variation), LSD = (least significant difference, MU = (Mekelle University)}

Similarly, recent research on groundnuts by Mohammed and Ismail [16] showed that plants that were inoculated with Rhizobium strain yielded a higher number of pods per plant while uninoculated plants yielded lower. This result is supported by the previous findings stating that Rhizobium inoculation enhances macro and micronutrient uptake and provides nitrogen to the host plant through biological nitrogen fixation [17].

Similar to pod number there was a significant (P < 0.05) variation in seed number per pod among the different biofertilizers, locations, and cropping seasons (Table 3). The application of biofertilizer significantly increased the seed number per pod as compared to the control and application of chemical fertilizer. The maximum number of seeds per pod (4.5) was counted in the integrated application of biofertilizer (Rhizobium and PSB) and NPKSZn fertilizer. The lowest number of seeds (2.6 and 2.9) was obtained in the control and application of NPKSZn respectively. The Atsela site produces significantly more pods per plant and seed per pod than the Mekelle site. Increased number of pod and seeds per pod in integrated application of Rhizobium and PSB along with NPKSZn could be due to increasing other nutrient absorption and in increasing available phosphorus of insoluble phosphorus sources. The result confirms the findings of Chimdi et al. [18] who also observed significant increment in number of seeds per pod when B. japonicum inoculation was combined with different levels of P. Similar results were reported by El-Gizawy et al. [19] and Yilmaz et al. [20] that P application and Rhizobium inoculation significantly increased pod formation, seed yield, and dry matter production in mung bean as compared with un-inoculated treatments. Jasim and Khudair [21] reported the highest seed yield (3.41 tons ha-1) observed in treatments using inorganic fertilizer (35 Kg ha-1 super phosphate triple + 100 Urea Kg ha-1) + nitrogen and phosphorus bio-fertilizers. In Ethiopia, inoculation of faba bean with rhizobia increased number of pods per plant from 7 to 13.08, nodules dry weight by 34.04%, 82% increase of nodules compared to un-inoculated plant [14].

3.6. Plant height

Analysis of variance for plant height revealed a significant difference among the different biofertilizers, location, and year (Fig. 2). Results revealed that inoculation of seed with Rhizobium and phosphate solubilizing bacteria increased plant height in comparison to control and application of chemical fertilizer. The longest plant height (148.8 cm) was recorded with dual inoculation of Rhizobium and PSB along with the application NPKSZn followed by seed inoculated with Rhizobium and PSB (138.4 cm) and application of Rhizobium along with the NPKSZn (132.5 cm). As compared to the control inoculation of seed with Rhizobium and PSB increased plant height by 27.2% and by 20.34% when it compared to the application of NPKSZn. Compared to the location and growing season the longest plant height was recorded at Atsela in the second year growing season. The present result was in accordance with Sahu et al. [22] who reported that inoculation of plants with Azospirillum could result in significant changes in various growth parameters, such as plant height. This result agreed with the result of Farfour [23] which affirmed that application of Rhizobium strains could increase the plant height of faba bean. Similarly, Molina-Romero et al. [24] showed that inoculation of maize seeds with all bacterial strains significantly increased plant height. Habtamu et al. [25] reported that combined application of NP fertilizer with Rhizobium inoculation provided the highest plant height. Raza et al. [26] and Sajid et al. [27] also found that Rhizobium inoculation increased the plant height of mung bean and ground nut, respectively.

Data in Table 4–8 show that significant seasonal effects (P < 0.05) existed for all characters studied except the number of branches and seeds per pod. Higher mean values for all characters were detected in the rain-fed season. It could be concluded that the increase in seed yield in the second season may be due to the significant increase in the soil microorganisms that increased the number of nodules and pods/plant and seed yield/plant.

Table 4

Analysis of variance for Biomass production rate, Biological yield, Economic growth rate, Growth production efficiency grain yield and harvest index of faba bean
SV	d.f.	TSW	BPR	BY	EGR	GPE	GY	HI
Block	2	240	14.2	136403	1.92	5.673	14262	67.92
Biof.	7	32708^**	214.1^**	5171549^**	736.70^**	350.69^**	26279^**	356.8^**
Loc.	1	16552^**	1414.4^**	28049653^**	717.00^**	10.51^*	11286^**	4329.2^**
Yr.	1	782 ^ns	55.08^**	3506331^**	4309.80^**	11329.8^**	53802^**	4472.1^**
Biof.^*Loc.	7	1715^**	4.50^ns	169395^**	4.36 ^ns	0.125 ^ns	155 ^ns	5.193 ^ns
Biof.^*Yr.	7	785*	105.9^**	537840^**	170.36^**	68.19^**	1944^**	120.2^**
Loc.^*Yr.	1	2562^**	50.9^**	3185776^**	25.50 ^ns	13.64^**	318 ^ns	1309.4^**
Biof.^*Loc.^*Yr.	7	418 ^ns	2.49 ^ns	3906 ^ns	1.01 ^ns	0.048 ^ns	11 ^ns	2.4 ^ns
Residual	62	320	3.026	37064.	12.66	1.506	154	8.809
CV		2.8	8.0	7.1	10.0	2.7	8.8	5.7
^{TSW = Thousand seed weight; BPR = Biomass production rate; BY = Biological yield; EGR = Economic growth rate; GPE = Growth production efficiency; GY = grain yield; HI = harvest index}

3.7. Thousand seed weight

Analysis of variance revealed that a significant difference was observed among the different biofertilizers. Compared to the control, the application of biofertilizer significantly increased the thousand seed weight of faba bean (Fig. 3). The highest thousand seed weight (699.9g and 674.5g) was recorded with dual inoculation Rhizobium and PSB along with the application of NPKSZn and application Rhizobium and PSB, respectively. The lowest seed weight (591.6g and 532.1g) was measured with the application of NPKSZn and control respectively. Among the locations, the highest seed weight was obtained at Atsela (671.15g). Dual inoculation of seeds with Rhizobium and PSB increased seed weight by 26.78, 14.01, 9, and 7.5% compared to control, application of NPKSZn, and sole application of Rhizobium and PSB respectively. This result was also in line with the work of Gedamu et al. [28] who reported that inoculation of faba bean with rhizobia strain alone could increase 100 seed weight. Bezabih et al. [14] reported that inoculation of faba bean with rhizobia increased hundred seed weight by 11.44%.

3.8. Grain yield (kg ha^-1)

The grain yield of faba bean was significantly (P < 0.05) affected by the main effect of biofertilizer, location, and year. Inoculation of seed with biofertilizer produced higher grain yield than control and application of chemical fertilizer. Dual inoculation of seed with Rhizobium and PSB along with NPKSZn gave the highest grain yield (2369 kg ha-1) followed by inoculation of faba bean with Rhizobium and PSB (1714 kg ha-1). The lowest grain yield (848 kg ha-1) was recorded at control followed by the application of chemical fertilizer NPKSZn (Table 6). Dual inoculation of seed with Rhizobium and PSB increased grain yield by 79.28 and 102.1% as compared to the application of chemical fertilizer (NPKSZn) and control respectively. The increase of seed yield under the influence of Rhizobium and PSB can be attributed to the ability of phosphate solution bacteria in biofertilizer to increase available phosphorus of insoluble phosphorus sources. Similarly, Ndlovu et al. [29] also reported about a 16.15–27.50% grain yield increment in two dry bean (Phaseolues vulgaris) cultivars due to Rhizobium inoculation. In another study, Rokhzadi et al. [30] reported that the seed yield of chickpea increased by application of biological fertilizers.

Table 5

Interaction effect of biofertilizer, location and growing season on growth and yield of fababbean
Treatment	PH				NN				NPPP				Gy (kg ha^− 1)
	Atsela		MU		Atsela		MU		Atsela		MU		Atsela			MU
	Off S.	R.F	Off S.	R.F	Off S.	R.F	Off S.	R.F	Off S.	R.F	Off S.	R.F		Off S.	R.F		Off S.	R.F
Control	88.00	136.0	88.00	123.3	45.33	47.33	42.00	43.67	9.67	20.33	11.67	17.00		505	1323		433	1134
NPKSZn	92.67	140.7	92.67	134.0	40.00	44.00	37.33	41.67	15.00	24.00	15.00	22.67		567	1493		486	1279
PSB	101.7	149.7	101.7	140.3	58.00	60.00	52.33	57.00	21.00	25.67	18.67	24.33		1156	1528		991	1309
PSB + NPKSZn	100.3	148.3	100.3	144.7	62.00	63.33	57.00	61.33	23.33	26.67	21.33	25.33		1202	1588		1030	1361
RZM	98.67	152.0	98.67	148.0	70.33	71.33	65.00	67.33	23.33	27.67	23.00	26.33		1389	1668		1191	1430
Rzm + NPKSZn	106.7	164.0	106.7	152.7	69.33	70.67	65.33	71.33	23.67	33.00	22.33	27.67		1642	1764		1408	1512
Rzm + PSB	113.3	171.0	113.3	156.0	79.67	81.00	76.67	79.67	25.33	36.00	24.33	31.00		1718	2007		1410	1720
Rzm + PSB + NPKSZn	119.0	188.7	119.0	166.3	79.67	81.00	76.67	81.00	27.67	44.00	25.00	39.67		2288	2814		1961	2412
LSD	6.867				2.37				2.378					202.2
^{PH = plant height; NN = number of nodules; NPPP = Number of pods per plant; Gy = Grain yield; MU = Mekelle University; Off S = off season; RF = rain fall}

3.9. Biological yield

Analysis of variance indicated biological yield was significantly (P < 0.05) affected by the application of biofertilizer. Maximum biological yield (3942 kg ha-1) was obtained by inoculation of faba bean with Rhizobium and PSB along with application of NPKSZn which was 125 and 95.92% higher as compared to control treatment (no chemical and biological fertilizers) and application of NPKSZn respectively (Table 3). Buernor et al. [31] and El-Habbasha et al. [32] findings also supported the results that the biological yield increase was higher in P-treated plants inoculated with biofertilizer than the plants inoculated with bacteria alone. Phosphate-solubilizing microorganisms had an important role in phosphorus solubility and uptake.

Table 6

Main effect of bio-fertilizers on yield and yield related traits of faba bean
Treatment	BY	BPR	TSW(g)	HI	GY (kg ha^− 1	GPE	EGR
Rzm + PSB + NPKSZn	3942^a	29.32 ^a	699.9 ^a	61.99 ^a	2369 ^a	53.41 ^a	51.36 ^a
Rzm + NPKSZn	2978^b	23.70^b	661.8^b	54.63^b	1582^c	46.25^c	39.36^b
RZM	2797^c	22.88^bc	627.2^c	52.13^c	1420 ^d	46.71 ^c	35.49^c
Rzm + PSB	2794 ^c	21.52 ^c	674.5^b	63.02^a	1714^b	50.16^b	39.87^b
PSB + NPKSZn	2669c	22.06 ^c	631.1^c	49.76^cd	1295^e	45.74 ^c	33.12^c
PSB	2657 ^c	22.32^bc	618.8 ^c	48.13^de	1246^e	41.67^d	34.83^c
NPKSZn	2012^d	17.12^d	591.6^d	47.12^e	956^f	38.41^e	28.60^d
Control	1750^e	15.38^e	532.1^e	46.37^e	848^g	38.08^e	25.07^e
LSD(5%)	157.1	1.42	14.60	2.486	101.1	1.01	2.893
CV(%)	7.1	8.0	2.8	5.8	8.7	2.7	9.9
Atsela	3240	25.63	671.15	46.22	1541	44.72	38.78
MU	2159	17.95	588.10	58.98	1317	45.38	33.15
Off season	2508	22.54	632.48	46.81	1211	55.92	29.78
Rain fed	2891	21.03	626.77	58.98	1646	34.19	42.23
LSD	78.6	0.71	7.30	1.243	50.5	0.501	1.446
^{BY = Biological Yield; BPR = Biomass production rate; TSW = thousand seed weight; HI = Harvest index; GPE = growth Production efficiency; EGR = Economic growth rate}

3.10. Harvest index

In the current investigation there was considerable variation in the percent ratio of economic and biological yield of the crop. Mean comparisons indicated that the maximum harvest index (63.02 and 61.99%) was obtained in dual inoculation of Rhizobium + PSB and inoculation of Rhizobium + PSB along with the application of NPKSZn respectively. While minimum harvest index (46.37%) was obtained by non-chemical and biofertilizer treatment (control). Similar results were shown by Buernor et al. [31] that indicated the harvested index was significantly affected by using chemical and biological phosphorus fertilizers. This is due to the variable rate of translocation of assimilate toward grain development, leading to variability in yield and HI value among the seven applied treatments [33].

3.11. Effect of different biofertilizer on Chocolate spot of faba bean

Chocolate spot is the most common serious disease that affects the production of faba bean. Analysis of variance clearly indicated that a significant difference (P < 0.001) was observed among the different biofertilizers for the number of infected plants per plot, Disease incidence, and disease severity index (Table 7).

Table 7

Analysis of variance for number of infected plant, disease incidence and disease severity index
Source of variation	d.f.	NIP	DI	DSI
Block	2	0.062	0.35	1.021
Treatment	7	148.76^**	843.32^**	82.56^**
Location	1	4.083^ns	23.15^ns	3.00^ns
Treatment.Location	7	1.988^ns	11.27^ns	7.95^ns
Residual	30	2.151	12.20	3.443
^{NIP = number of infected plant per plot; DI = Disease incidence; DSI = disease severity index}

The current result for the incidence of chocolate spots indicated that the application of biofertilizer significantly reduced the incidence. The lowest number of infected plants per plot (5.67 and 9.17) with disease incidence level of (13.49 and 21.83%) was achieved with the combined application of Rhizobium + PSB and NPKSZn and plots treated with dual inoculation of Rhizobium + PSB respectively (Table 8). The highest number of infected plants (21.33) with an incidence level of (50.79%) was obtained with the application of chemical fertilizer (NPKSZn). Chandramani et al. [34] revealed that the combination of FYM, three biofertilizer, lignite fly ash, and neem cake applied in split doses significantly reduced the incidence leaf folder (76.69%), stem borer (58.66), and gall midge (66.81%) as compared to NPK applied as inorganic form on paddy. Similar results were observed by Chatterjee et al. [35] where different sources of nutrients had a significant effect on reducing the whitefly population and the treatments containing higher amounts of FYM or vermin-compost showed better results over sole inorganic fertilizers.

3.12. Chocolate spot severity index

Data presented in Table (8) showed that dual inoculation of seed with Rhizobium and PSB significantly reduced Chocolate spot severity index compared to single inoculation, control, and application of chemical fertilizer (NPKZn). The lowest Chocolate spot severity index (13.33 and 16.67%) was recorded in dual inoculation of seed with Rhizobium and PSB along with and without application of chemical fertilizer (NPKSZn) respectively. On the other hand, the highest Chocolate spot severity index (25.17 and 23.17%) was observed with the application of chemical fertilizer (NPKSZn) and control respectively (Table 8).

Table 8

Effect of different biofertilizer on chocolate spot of faba bean
Treatment	NIP	DI (%)	DSI (%)
NPKSZn	21.33^a	50.79^a	25.17^a
Control	18.50^b	44.05^b	23.17^a
RZM	15.17^c	36.11^c	19.67^bc
PSB + NPKSZn	15.00^c	35.71^c	20.17^b
RZM + NPKSZn	15.00^c	35.71^c	17.50^cd
PSB	12.17^d	28.97^d	20.00^b
RZM + PSB	9.17^e	21.83^e	16.67^d
RZM + PSB + NPKSZn	5.67^f	13.49^f	13.33^e
Pvalue	< 0.001	< 0.001	< 0.001
SEm±	0.599	1.426	0.758
Atsela	14.04	32.64	19.21
MU	14.46	34.03	19.71
SEm±	0.299	0.713	0.379
CV	10.5	10.5	9.5
^{NIP = number of infected plant per plot; DI = Disease incidence; DSI = Disease severity index; SEm = Standard error of the mean}

3.13. Partial Budget Analysis

According to the results of partial budget analysis, the highest net benefit was obtained from the application of Rhizobium and PSB inoculation along with the application of NPKSZn fertilization (ETB 48174.9 ha-1) followed by dual inoculation of Rhizobium and PSB (ETB 35919.4 ha-1) and Rhizobium inoculation along with the application of Rhizobium and chemical fertilizer NPKSZn (ETB 31462.2 ha-1) (Table 9). As compared to the control, inoculation of seed with Rhizobium and PSB along with the application of NPKSZn; Rhizobium and PSB sole inoculation of Rhizobium and PSB increased the net benefit by 161.25, 94.78, 63.23 and 42.38% respectively (Table 9).

Table 9

Partial budget analysis for an experiment on different biofertilizer combination of faba bean
Treatment	Average yield (kg/ha)	Adjusted yield (kg/ha)	Gross field benefits.(ETB/ha)	Total variable cost (ETB/ha)	Net Benefit (ETB/ha)	Dominance analysis
Rzm + PSB + NPKSZn	2369	2013.65	52354.9	4180	48174.9	Non dominated
Rzm + NPKSZn	1582	1344.7	34962.2	3500	31462.2	Non dominated
RZM	1420	1207	31382	1280	30102	Non dominated
Rzm + PSB	1714	1456.9	37879.4	1960	35919.4	Non dominated
PSB + NPKSZn	1295	1100.75	28619.5	3500	25119.5	Non dominated
PSB	1246	1059.1	27536.6	1280	26256.6	Non dominated
NPKSZn	956	812.6	21127.6	2520	18607.6	Non dominated
Control	848	720.8	18740.8	300	18440.8
^{ETB= Ethiopian Birr}

3.14. Marginal Rate of Return Analysis (MRRA)

Based on the dominance analysis all the treatments were included for further Marginal Rate of Return analysis (Table 10). For a treatment to be considered a worthwhile option for farmers, the marginal rates of return (MRR) need to be at least between 50% and 100% [10]. Thus, to draw recommendations from marginal analysis in this study, a 100% return to the investment is a reasonable minimum acceptable rate of return for farmers in the study area. Accordingly, the marginal rate of return NPKSZn was 12.56% which is not within the acceptable range.

From the other treatment seed inoculated with Rhizobium had the highest marginal rate of return (1336.5%) followed by dual inoculation of Rhizobium and PSB (1126.83% MRR), PSB (899.52%) and combination of Rhizobium and PSB along with the application of NPKSZn (789.26%). This implies that for Birr 1.0 investment in faba bean production, the producer can get Birr 133.65, 112.68, 89.95, and 78.92 respectively. The lowest but above the acceptable range of marginal rate of return (218.67%) was obtained in seeds inoculated with PSB and NPKSZn followed by seeds inoculated with Rhizobium and NPKSZn (Table 10). This implied that seed inoculation with Rhizobium and PSB could produce higher yields and a high rate of return for farmers in Atsela and similar agro-ecology.

Table 10

Marginal rate of return based on the grain yield and current price of faba bean
Treatment	Adjusted yield (kg ha^-1)	Gross field benefits.(ETB ha^-1)	Total variable cost (ETB ha^-1)	Net Benefit (ETB ha^-1)	MRR
Rzm + PSB + NPKSZn	2013.65	52354.9	4180	48174.9	789.26
Rzm + NPKSZn	1344.7	34962.2	3500	31462.2	423.27
RZM	1207	31382	1280	30102	1336.50
Rzm + PSB	1456.9	37879.4	1960	35919.4	1126.83
PSB + NPKSZn	1100.75	28619.5	3500	25119.5	218.67
PSB	1059.1	27536.6	1280	26256.6	899.52
NPKSZn	812.6	21127.6	2520	18607.6	12.58
Control	720.8	18740.8	300	18440.8
^{MRR = Marginal Rate of Return}

In light of the obtained results, it could be concluded that under the conditions during this investigation, enrichment of faba bean plants with dual inoculation of Rhizobium, PSB combined with the application of NPKSZn gave a considerable increase in yield and yield components as well as reduced number of infected plant and severity of chocolate spot. The lowest number of infected plants, disease incidence level, and lowest Chocolate spot severity index were achieved with combined application of Rhizobium + PSB and NPKSZn and plots treated with dual inoculation of Rhizobium + PSB respectively. In contrast, the highest number of infected plants, incidence, and severity index were obtained with the application of chemical fertilizer (NPKSZn). The highest net benefit was also obtained from the application of Rhizobium and PSB inoculation along with the application of NPKSZn fertilization and dual inoculation of Rhizobium and PSB. The application of biofertilizer is an acceptable approach for higher yield with good quality and safe for human consumption. The current results show that either single or mixed inoculation gave a positive response to the studied parameters. This response was accompanied by a significant increase in plant height, pod number, seed number per pod, thousand seed weight, and biological yield.

Competing Interests

The authors declare that there are no financial or non-financial competing interests regarding the publication of this article.

Funding

The work was financially supported by Mekelle University with the Grant project Registration Number of CRPO/ CDANR /Medium/ Recurrent/001/2011.

Author Contribution

All authors contributed to the study's conception and design. Material preparation, data collection, and analysis were performed by [Mohammed Mebrahtu Mossa], [Daniel Gebrekidan], and [Etsay Mesele]. The first draft of the manuscript was written by [Mohammed Mebrahtu Mossa] and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Data Availability

The datasets generated during and/or analyzed during the current study are available in the manuscript.

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You are reading this latest preprint version

Effectiveness of different bio-fertilizer on growth, yield and yield attributing character of Faba bean (Vicia fabae L.)

Status:

Version 1

Abstract

Figures

1. Introduction

2. Material and methods

2.1. Description of the experimental site

2.2. Treatment and Experimental Design

2.3. Data Collection

2.4. Data analysis:

2.5. Partial Budget Analysis

3. Result and discussion

3.1. Soil Physical properties of the experimental site

3.2. Effect of different biofertilizer on phonological traits of faba bean

3.3. Days to Flowering, Maturity and grain filling period

3.4. Number of branches and nodule number

3.5. Number of pod per plant and Seeds per pod

3.6. Plant height

3.7. Thousand seed weight

3.8. Grain yield (kg ha^-1)

3.9. Biological yield

3.10. Harvest index

3.11. Effect of different biofertilizer on Chocolate spot of faba bean

3.12. Chocolate spot severity index

3.13. Partial Budget Analysis

3.14. Marginal Rate of Return Analysis (MRRA)

4. Conclusion

Declarations

Competing Interests

Funding

Author Contribution

Data Availability

References

Additional Declarations

Status:

Version 1

Effectiveness of different bio-fertilizer on growth, yield and yield attributing character of Faba bean (Vicia fabae L.)

Status:

Version 1

Abstract

Figures

1. Introduction

2. Material and methods

2.1. Description of the experimental site

2.2. Treatment and Experimental Design

2.3. Data Collection

2.4. Data analysis:

2.5. Partial Budget Analysis

3. Result and discussion

3.1. Soil Physical properties of the experimental site

3.2. Effect of different biofertilizer on phonological traits of faba bean

3.3. Days to Flowering, Maturity and grain filling period

3.4. Number of branches and nodule number

3.5. Number of pod per plant and Seeds per pod

3.6. Plant height

3.7. Thousand seed weight

3.8. Grain yield (kg ha-1)

3.9. Biological yield

3.10. Harvest index

3.11. Effect of different biofertilizer on Chocolate spot of faba bean

3.12. Chocolate spot severity index

3.13. Partial Budget Analysis

3.14. Marginal Rate of Return Analysis (MRRA)

4. Conclusion

Declarations

Competing Interests

Funding

Author Contribution

Data Availability

References

Additional Declarations

Status:

Version 1

3.8. Grain yield (kg ha^-1)