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Article

Yield and Seed Quality of Faba Bean (Vicia faba L. var. minor) as a Result of Symbiosis with Nitrogen-Fixing Bacteria

by
Magdalena Serafin-Andrzejewska
*,
Agnieszka Falkiewicz
,
Wiesław Wojciechowski
and
Marcin Kozak
Institute of Agroecology and Plant Production, Wrocław University of Environmental and Life Sciences, 24A Grunwaldzki Square, 50-363 Wrocław, Poland
*
Author to whom correspondence should be addressed.
Agriculture 2025, 15(9), 960; https://doi.org/10.3390/agriculture15090960 (registering DOI)
Submission received: 14 March 2025 / Revised: 24 April 2025 / Accepted: 24 April 2025 / Published: 28 April 2025
(This article belongs to the Special Issue Advances in the Cultivation and Production of Leguminous Plants)
Versions Notes

Abstract

:
Faba bean is a high-protein legume that can be successfully grown in most climates around the world. It is one of the most popular pulses cultivated in Poland. Its seeds are a source of plant protein, used most often in feed production. Field experiments and laboratory seed analyses were carried out in 2022 and 2023 to assess the effect of the application of nitrogen-fixing bacteria on the yield and seed quality of a low-tannin faba bean cultivar. The factor was tested at four levels: control, seed inoculation with Rhizobium leguminosarum bv. viceae, foliar spraying with Methylobacterium symbioticum, and seed inoculation and spraying (double application). The application of N-fixing bacteria had a positive effect on faba bean seed yield. In 2022, plants responded most effectively to a double application, increasing seed yield by 25.4%, while, in 2023, the highest seed yield was obtained after inoculation (12.3% increase). Although the single application of bacteria caused a decrease in seed protein content, the double application (inoculation and spraying) significantly enhanced seed protein content. The protein productivity per hectare was compensated by the higher seed yield and increased by 41.7% in 2022 and 14.9% in 2023 compared to plots where N-fixing bacteria were not applied. This work shows that it is possible to use different strains of N-fixing bacteria in faba bean cultivation and this can significantly improve yields while reducing the need for synthetic nitrogen fertilizers, which supports sustainable production.

1. Introduction

For many years, the European Union (EU) has been forcing member states to increase the area under legumes. The main purpose of promoting pulses is to diversify sources of both consumer and feed protein, which is currently mainly imported. The high protein content of legumes is one of the main factors determining the economic importance of these crops. Among all crops, legumes contain the most protein, which has a high biological value. Depending on the species and growing conditions, this value ranges from 20 to 42%; in comparison, in cereal grains, the content does not exceed 18% [1].
Faba bean (Vicia faba L. var. minor), also known as horse bean, broad bean, or fava bean, is cold-tolerant and does well in cool and wet regions, making it a very attractive legume in cold climates where soybeans will not thrive and yield well [2].
Globally, faba bean is grown mainly in China, Europe, the northern part of Africa, West Asian countries, and Australia [3]. In 2023, its global area under cultivation was nearly 2.8 million hectares, with dry seed production of above 6 million tons. World faba bean seed production can be considered stable over the past few years [4].
According to Eurostat [5], in the European Union, in 2022, faba bean was grown on 436,000 hectares, of which 33,000 hectares were located in Poland. In 2019, the average yield in the European Union was 2.90 t ha−1 and, in Poland, there was 2.30 t ha−1 of dry faba bean [4]. Data on the production of grain legumes in 2014–2018 in the EU show that faba bean production with about 1.8 million t is the third most cultivated legume after soybeans and dry peas. In the European Union, faba bean seeds are mainly used as feed. Exports outside the EU are mainly to two markets: Egypt as food and Norway as fish feed. Poland is the eighth producer of faba bean seeds in the EU [6].
A great benefit of legumes is their ability to symbiose with atmospheric nitrogen-fixing bacteria. The finding of nitrogen-fixing microorganisms and a better understanding of the Biological Nitrogen Fixation (BNF) process has led to the creation of many biopreparations around the world. As early as 1895, seed inoculant produced from Rhizobium strains was introduced to the market in the United States [7]. Today, a number of biopreparations containing nitrogen-fixing bacteria are used, showing beneficial effects in improving yields as well as reducing production costs.
BNF allows legumes to fix significant amounts of atmospheric N2, which contributes to minimizing the use of synthetic nitrogen fertilizers and improves soil conditions and biodiversity [8]. It is estimated that between 139 and 170 million tons of N enter the global nitrogen cycle annually and about 70–80% comes from the nitrogen-fixing process of symbiotic bacteria [9]. Although Rhizobium bacteria usually naturally occur in Polish soils, a common practice is pre-sowing inoculation of seeds [10]. Each legume species coexists with strictly selected legume bacteria, while faba bean maintains a symbiosis with the bacterium Rhizobium leguminosarum bv. viceae. Through the cultivation of faba bean during one growing season, we can introduce up to 200 kg N ha−1 into the soil. Unfortunately, the amount of bound nitrogen is strongly dependent on the environmental conditions in the field. One of the popular agronomic treatments for improving the efficiency of the BNF process is a thorough covering of seeds with bacteria. A lesser-known way to increase N2 reduction is foliar spraying of plants with endophytic bacteria. Regardless of the method of application of nitrogen-fixing bacteria, the BNF process makes it possible to reduce the use of nitrogen fertilizers, thus cutting production costs, as well as environmental pollution [11].
Biological products based on strains of Methylobacterium symbioticum bacteria may become a new issue for BNF. These bacteria, unlike other atmospheric nitrogen fixers, colonize not the root zone but the aboveground part of plants. The product’s action is based on its penetration through the stomata into plant tissues, near photosynthesizing tissues [12]. It provides the plant with a supply of nitrogen at every stage of growth. Symbiotic activity has been demonstrated not only to legumes, but to a very wide spectrum of species. The first studies were conducted on grain corn and rice, as well as grapevine [13]. Symbiosis has also been demonstrated with cereal crops, rapeseed, potato, and sugar beet but also in many vegetable or fruit crops. However, the existing data in the literature are sparse and often relate to a single growing season [14] or suggest further research to understand the specific action of the bacteria [15], which is insufficient for inferring the efficacy of preparations containing M. symbioticum.
Due to the low popularity of legume cultivation in Poland, there are not many of Rhizobium bacteria in the soils, especially when the plant is not regularly present in the crop rotation. It is therefore important to inoculate the seeds or, as presented in this study, to look for other strains of N-fixing bacteria, such as those applied by foliar spraying. Therefore, efforts are being made to search for the best methods of bacterial application so that the seed yields of native Polish legumes, represented in this study by faba bean, are stable and satisfactory not only in terms of quantity but also quality. This became the immediate premise for the work on this topic.
The main purpose of this study was to determine the effect of the application of preparations containing diverse atmospheric nitrogen-fixing bacteria on faba bean seed yield and protein content.

2. Materials and Methods

2.1. Field Experiment Location and Design

A field experiment was carried out in 2022 and 2023 to evaluate the effect of using N-fixing bacteria on the yield and protein content of faba bean seeds. The trial was located in the fields of the Wrocław University of Environmental and Life Sciences in Poland (51°10′ N, 17°06′ E).
Domino cultivar was used for testing. It is an indeterminate cultivar, characterized by low tannin content in seeds (the average content for 2023–2024 was 0.0103 mg g−1 in dry matter) [16], suitable for growing for seeds. Seed and protein yields are quite high. Date of flowering and maturity are slightly later than average, and the flowering period is quite long. The plants are tall and resistance to lodging at the end of flowering is high, while, before harvest, it is medium. The weight of 1000 seeds is very low. The content of total seed protein and crude fiber is quite high. The cultivar is suitable for growing on soils of wheat complexes. The optimum sowing density is about 50 seeds per 1 m2 [17].
The field experiments had a randomized complete block design (RCBD) with one factor: the use of N-fixing bacteria.
  • Two commercial products were selected for testing:
  • An inoculant for faba bean, containing Rhizobium leguminosarum bv. viciae (applied by seed inoculation);
  • A product containing Methylobacterium symbioticum (applied according to the producer’s recommendations through foliar spraying);
  • As a third variant, both treatments were applied;
  • Objects without bacterial application (control) were also included for comparison.
  • This provided four variables to test in the trial.
The experiment consisted of four randomized replicates. The area of a single experimental plot was 15 m2 (10 m × 1.5 m).

2.2. Soil and Weather Conditions

The field trial was located on Cutanic Stagnic Luvisol—developed from light loam underlain by medium loam. It is a soil suitable for growing wheat [18,19].
The following chemical analyses were carried out in the soil samples taken immediately prior to the field experiment:
  • Soil pH—in 1 mol dm−3 KCl—potentiometrically;
  • Available phosphorus content—Egner–Riehm spectrophotometric method;
  • Available potassium—flame photometry method;
  • Bioavailable magnesium—Schachtschabel method.
The pH of the soil was neutral both in 2022 and in 2023. The soil was characterized by very high P content in 2022 and in 2023, high (2022) or very high (2023) K content, and very low Mg content in both years of the research [20] (Table 1).
Weather conditions during the study years and the average temperatures and precipitation from 1991 to 2020 are shown in Table S1. The data come from a weather station located exactly in the experimental fields.

2.3. Agrotechnical Management

In the field experiment area, mineral fertilization was applied immediately before sowing at the following rates (kg ha−1): 60 P2O5 (46% triple superphosphate) and 120 K2O (60% sylvinite). The fertilizers were then mechanically mixed into the soil to a depth of about 5 cm. In each year of the field experiment, the forecrop was winter wheat.
Directly before sowing, part of the seeds was inoculated with a faba bean seed inoculant containing R. leguminosarum bv. viciae. Seeds were sown on plots (15 m2) according to the field experiment design.
For determining faba bean growing stages, a Biologische Bundesanstalt, Bundessortenamt und Chemische Industrie (BBCH) scale was used.
Spraying faba bean plants with M. symbiothicum was carried out on plots (15 m2) according to the experiment design at BBCH 13 growing stage.
The detailed cultivation of faba bean is shown in Table 2 for each year of the study separately.

2.4. Data Collection and Chemical Analyses of Seeds

The components of yield were determined right before harvest (BBCH 86–87) on a sample of 10 plants from the central part of each plot. The following traits were measured: pod number per plant, seed number per plant, and seed weight per plant. After harvest, the 1000-seed weight from each plot according to the International Rules for Seed Testing [21] and the seed yields were determined. A moisture content of 15% was used to convert seed yields from plots to seed yields per hectare.
Protein content in harvested seeds was established with a modified Kjeldahl method, which involves combustion or mineralization of nitrogenous compounds, followed by distillation and titration. When converting total nitrogen to total protein, the coefficient 6.25 was used.
To calculate the total protein productivity, dry matter was measured, using the gravimetric method (105 ± 2 °C for 5 h).
Based on dry matter, total protein content, and the seed yield, the total protein productivity was calculated from 1 ha.

2.5. Statistical Analyses

According to Lindeberg-Lévy’s theorem (central limit theorem, CLT), the distribution of variables such as biometric, quantitative, and qualitative traits of crop converge asymptotically to the normal distribution.
The analysis of variance (ANOVA) was performed, and the significance levels were α ≤ 0.001, α ≤ 0.01, and at α ≤ 0.05, using the program Statistica 13.1 (StatSoft, Kraków, Poland) for each year of the study separately. The experiment included one factor: four applications of N-fixing bacteria (inoculation with R. leguminosarum bv. viciae, spraying with M. symbiothicum, inoculation + spraying, and control) and four randomized replicates set as an RCBD. For the resulting data on the traits studied, standard deviations (SD) were calculated. In order to identify homogeneous groups, a Tukey’s multiple-range test was performed (at α ≤ 0.05), using successive letters to describe them, starting from ‘a’—the most favorable to ‘d’—the least favorable in terms of the values of the analyzed characteristics.

3. Results

Analysis of variance showed that the application of N-fixing bacteria had a strong impact on all analyzed characteristics in both years of research, except for the number of pods per plant in 2023 (Table 3).
In 2022, the effect of the application of symbiotic bacteria was demonstrated in number and weight of seeds per plant and number of pods per plant (Table 4). Inoculation or spraying, as well as double treatment, resulted in significantly higher values of the traits studied, compared to control.
The number of seeds harvested per plant increased by 16.6% after the application of both bacterial preparations (inoculation + spraying), compared to the control.
Application of R. leguminosarum bv. viciae (inoculation) resulted in an 18.8% increase in seed weight obtained from a single plant, while the double treatment led to an increase of 25.1%, compared to the control.
The greatest number of pods per plant was produced by plants treated with M. symbiothicum (spraying) and this was 13.4% greater than on objects where the bacteria were not applied at all (Table 4).
Compared to the control, the application of bacterial products had a significant effect on thousand-seed weight (Table 5). Application of R. leguminosarum bv. viciae (inoculation) increased seed mass by 5.4%, while spraying M. symbiothicum increased seed mass by 8.9%.
The highest seed yield (25.4% higher than the control) was obtained from plots with a double application of N-fixing bacteria, followed by those with a single application, and the lowest yield was obtained from control plots (Table 5).
A decrease in total protein values was observed after using N-fixing bacteria (inoculation or spraying) compared to the control. The double treatment (inoculation + spraying) resulted in significantly the highest total protein content in the seeds, 4.1% higher compared to the objects where no bacterial preparations were used.
Considering total protein productivity, its value was clearly compensated by seed yield. The highest value for this trait was recorded after using R. leguminosarum bv. viciae and M. symbiothicum (double application), followed by a single application of the bacteria with no significant differences between the strain used, while the lowest value was recorded on control objects. The difference between the highest protein productivity and the lowest was 41.7% (Table 5).
In 2023, the application of nitrogen-fixing bacteria (both ways separately and double treatment) had a significant effect on all traits tested, except for the number of pods per plant (Table 6). In the second year of the study, it was shown that the tested applications of N-fixing bacteria had a more significantly differentiated effect on the studied traits of faba bean.
The bacterial products used had very similar effects on the number and weight of seeds per plant. The application of M. symbiothicum, as well as double treatment consisting of inoculation with R. leguminosarum bv. viciae and spraying with M. symbiothicum, resulted in the best values for both traits analyzed. After M. symbiothicum spraying, the number of seeds increased by 22.4% and their weight by 14.9%, compared to the control objects.
The most beneficial effect on the weight of 1000 seeds came from M. symbiothicum spraying and the dual application of N-fixing bacteria (inoculation and spraying). There were no differences between seed weights from plots inoculated with R. leguminosarum bv. viciae and from control plots (Table 7).
The highest seed yield was obtained after inoculation with R. leguminosarum bv. Viciae, as well as after double treatment (inoculation + spraying), then significantly lower after M. symbiothicum spraying, and the lowest when application was not performed. The difference between the highest and lowest seed yields was 12.3%.
As in 2022, the lowest total protein content in seeds was observed after inoculation or spraying, followed by the control, and the highest value of this trait was recorded with double treatment (inoculation + spraying).
Double application of symbiotic bacteria resulted in the highest total protein productivity, 14.9% higher than from control plots. Inoculation gave better results than spraying, considering the value of this trait. The lowest protein yield was obtained from the control objects (Table 7).
In both years of the study, a negative relationship can be observed between total protein content in seeds and seed yield. A single application of nitrogen-fixing bacteria, neither inoculation with R. leguminosarum bv. viciae nor M. symbiothicum spraying, broke this relationship. Only two treatments (inoculation + spraying) were able to cause high values for both traits considered.
In general, the best effect on all analyzed quantitative and qualitative traits of faba bean was shown for double treatment, both seed inoculation with R. leguminosarum bv. viciae and foliar spraying with M. symbiothicum.

4. Discussion

The efficiency of symbiotic nitrogen fixation (BNF) depends on many factors. The amount of fixed nitrogen is influenced by soil conditions, plant species, genetic characteristics of plants and symbiotic bacteria, as well as agrotechnical treatments carried out [22]. Many scientists are studying the effect of using nitrogen-fixing bacteria on the yield of legumes [23,24,25,26]. Unfortunately, in the available literature, there is a lack of studies examining how the method of bacterial application and its type affect legumes.
Poland is located in a humid continental climate area. In 2022, the average air temperature was 9.5 °C, 0.8 degrees higher than the annual average for the multi-year period (1991–2020). In 2023, the average annual temperature for the country was 10 °C and it was the second warmest year of the 21st century. The average annual amount of rainfall in Poland is currently 500–600 mm, but its spread is uneven over the country’s area and over the year [27]. It is assumed that, under Polish conditions, the application of bacterial inoculation is not always necessary, especially in fields where the bean crop species is grown regularly [10]. It has been found that there is no need for seed inoculation if suitable bacterial strains are present in the soil [28,29].
In the experiment conducted, in both years, the effect of nitrogen bacteria application on the number of seeds per plant and seed weight per plant was noted. In 2022, the increase in the number of seeds per plant, relative to the control object, was best influenced by seed inoculation, as well as double application of bacteria (inoculation + spray), while the increase in seed weight per plant was most favorably influenced by spray and with the same effect by double application. In 2023, the best effects for both traits in question were observed after spraying and double application. Gedamu et al. [30] also confirmed the effect of Rhizobium bacteria on a higher number of seeds per plant. However, there are also studies showing no effect of seed inoculation with bacteria on the number of seeds per plant [31].
After the application of bacteria in 2022, the number of pods per plant increased compared to plants from the control, and spraying was the most beneficial, as well as double application (inoculation + spraying). On the other hand, in 2023, the application of symbiotic bacteria had no effect on the trait in question. A study by Gedamu et al. [30] confirms the positive effect of inoculation on the number of pods per plant in protein crops. A similar result was shown for peas, when seed inoculation had a positive effect on pod number, doubling the number of pods compared to plants from a control plot [25].
The application of nitrogen-fixing bacteria increased the yield of faba bean seeds, regardless of the application technique. However, the best results were obtained in 2022 after double application and in 2023 after seed inoculation on an equal footing with double application. Denton et al. [23] also showed a beneficial effect of inoculation with Rhizobium bacteria on crops yield. According to Youseif et al. [24], after the application of effective strains, the yield of faba bean seeds increased by more than 40% relative to plants from the control object. Information on the effect of foliar spraying with M. symbioticum bacteria is, for the time being, difficult to access due to the innovation of this treatment. Witt and Weaver [14] compared soybean yields using spraying with nitrogen-fixing bacteria. They showed no significant differences between control, fungicide application, and spray application. However, it should be noted that the cited data are for only one growing season.
A single application of bacteria, seed inoculation, as well as foliar spraying, resulted in a decrease in protein content compared to the control object, but the use of a double application (inoculation + spray) gave significantly the highest total seed protein content. Such a phenomenon was observed in both years of the study. In our opinion, fixed nitrogen was used to accumulate dry matter and promote yield. Our conclusion is based on the study by Rodelas et al. [32,33]. In fact, in our study, only a double application of symbiotic bacteria resulted in an increase in the protein content of the seeds.
The connection between seed yield and protein content in legumes is an important issue and widely studied. Some papers report a positive relationship between the quantity and quality of faba bean seeds [34,35], but others inform of a negative relation between seed yield and protein content [36,37,38], which is in agreement with our research. This can be caused by a variety of biotic and/or abiotic factors, but climate, particularly temperature and precipitation distribution during the growing season, is considered to be the most influential factor in the variability of yield and quality of faba bean seeds [39,40]. In our research, this relationship was broken by the double application of the bacteria, both seed inoculation and foliar spraying, which perhaps proves the validity of performing these two treatments in faba bean cultivation.
Protein productivity is the most important indicator of crop success and profitability for producers. It is a value that depends on the yield of seeds and their protein content. In our study, the highest protein yield was obtained using double application of bacteria, followed by single treatments, and the lowest on control objects. The magnitude of this trait was more influenced by seed yield than by its protein content. However, the study showed the usefulness of the application of symbiotic bacteria in faba bean cultivation as a treatment to increase protein productivity, and it is most beneficial to both inoculate the seeds and apply a foliar spraying.
The discussion of the topic undertaken is difficult because there are practically no scientific studies available on the effects of foliar application (spraying) of atmospheric nitrogen-fixing bacteria M. symbioticum on faba bean plants, and the research on the response of faba bean to such a treatment undertaken in this paper is likely to be novel.

5. Conclusions

The use of products based on N-fixing bacteria had a significant effect on faba bean productivity. The synergistic effect of seed inoculation with R. leguminosarum bv. viciae and foliar spraying with M. symbioticum proved to be most beneficial for seed yield, protein content, and protein productivity, the traits on which the success of faba bean cultivation is mainly based.
The use of a novel bacterial strain, M. symbioticum, as a foliar spraying had at least as favorable an effect on faba bean yield parameters as seed inoculation with R. leguminosarum bv. viciae. This proves that spraying with this strain can be as effective as commonly used seed inoculants.
To summarize, application of N-fixing bacteria in faba bean cultivation is an effective and ecological method of supporting agriculture, which is conducive to both increasing protein yields and protecting the environment and should be widely recommended and implemented in agricultural practice.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/agriculture15090960/s1, Table S1: Weather conditions during the years of field experiment.

Author Contributions

Conceptualization, M.S.-A. and M.K.; methodology, M.S.-A., M.K. and A.F.; formal analysis, M.S.-A., A.F., W.W. and M.K.; investigation, M.S.-A. and M.K.; writing—original draft preparation, M.S.-A.; writing—review and editing, M.K. and W.W.; visualization, W.W.; supervision, M.K. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

Not applicable.

Data Availability Statement

The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding author.

Acknowledgments

The authors would like to thank Corteva Agriscience Poland Ltd. for donating the bacterial preparations for the study.

Conflicts of Interest

The authors declare no conflicts of interest. The donors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

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Table 1. Soil properties in years of research (pH and content of macronutrients).
Table 1. Soil properties in years of research (pH and content of macronutrients).
YearpHContent of Macronutrients
(mg 1000 g−1)
PKMg
20227.0226.114012.2
20236.8015.43028.4
Table 2. The detailed faba bean cultivation in 2022 and 2023.
Table 2. The detailed faba bean cultivation in 2022 and 2023.
CultivationYear
20222023
DateDetailsDateDetails
Sowing14 AprilRow spacing 15 cm;
60 seeds per 1 m2		29 MarchRow spacing 15 cm;
60 seeds per 1 m2
Weeds control14 AprilBoxer 800 EC (3.2 L ha−1, active substance: prosulfocarb)29 MarchBoxer 800 EC (3.8 L ha−1, active substance: prosulfocarb)
25 MayBasagran 480 SL (2.5 L ha−1, active substance: bentazone)17 MayBasagran 480 SL (1.0 L ha−1, active substance: bentazone) + Inex-A (0.05 L ha−1, adjuvant)
Spraying with
M. symbioticum		10 MayBBCH 138 MayBBCH 13
Insects (aphids) control13 MayLos Ovados 200 SE (0.1 L ha−1, active substance: acetamipiride) + Spaviero 100 CS (0.1 L ha−1, active substance: lambda-cyhalothrin)17 MayDelmetros 100 SC (0.05 L ha−1, active substance: deltamethrin)
+ Los Ovados 200 SE (0.25 L ha−1, active substance: acetamipiride)
19 MayKarate Zeon 050 CS (0.15 L ha−1, active substance: lambda-cyhalothrin)22 MayTop Gun 050 CS (0.1 L ha−1, active substance: lambda-cyhalothrin)
7 JuneDelmetros 100 SC (0.05 L ha−1, active substance: deltamethrin) + Los Ovados 200 SE (0.2 L ha−1, active substance: acetamipiride)31 MayTop Gun 050 CS (0.1 L ha−1, active substance: lambda-cyhalothrin)
24 JuneLos Ovados 200 SE (0.2 L ha−1, active substance: acetamipiride)
12 JulyLos Ovados 200 SE (0.2 L ha−1, active substance: acetamipiride)
Harvesting8 AugustBBCH 9023 AugustBBCH 90
Table 3. The impact of the application of N-fixing bacteria on quantitative and qualitative traits of faba bean in 2022 and 2023—ANOVA.
Table 3. The impact of the application of N-fixing bacteria on quantitative and qualitative traits of faba bean in 2022 and 2023—ANOVA.
Trait20222023
α–ValueSignificanceα–ValueSignificance
Number of seeds per plant0.0002***<0.0001***
Weight of seeds per plant (g)<0.0001***0.0001***
Number of pods per plant0.0040**0.0685ns
1000-seed weight (g)<0.0001***0.0003***
Seed yield (Mg ha−1)<0.0001***<0.0001***
Total protein content (g kg−1)<0.0001***<0.0001***
Total protein productivity (kg ha−1)<0.0001***<0.0001***
*** significant at α ≤ 0.001, ** significant at α ≤ 0.01, ns—not significant.
Table 4. The impact of the application of N-fixing bacteria on yield components of faba bean in 2022.
Table 4. The impact of the application of N-fixing bacteria on yield components of faba bean in 2022.
ApplicationNumber of Seeds per PlantWeight of Seeds per Plant (g)Number of Pods per Plant
R. leguminosarum bv. viciae (inoculation)24.65 ab ± 0.9111.44 b ± 0.378.00 b ± 0.22
M. symbiothicum (spraying)24.20 b ± 0.2911.51 ab ± 0.088.45 a ± 0.44
Inoculation + spraying25.63 a ± 0.4312.05 a ± 0.368.25 ab ± 0.31
Control21.98 c ± 0.579.63 c ± 0.317.45 c ± 0.26
Different letters indicate a significant difference (Tukey’s multiple range test). ± means SD.
Table 5. The impact of the application of N-fixing bacteria on 1000-seed weight, seed yield, total protein content, and productivity of faba bean in 2022.
Table 5. The impact of the application of N-fixing bacteria on 1000-seed weight, seed yield, total protein content, and productivity of faba bean in 2022.
Application1000-Seed Weight (g)Seed Yield
(Mg ha−1)		Total Protein Content
(g kg−1)		Total Protein Productivity
(kg ha−1)
R. leguminosarum bv. viciae (inoculation)460.25 b ± 1.713.11 b ± 0.02291.2 c ± 1.56768.5 b ± 5.45
M. symbiothicum (spraying)475.00 a ± 2.583.09 b ± 0.10293.7 c ± 2.79770.5 b ± 24.24
Inoculation + spraying469.75 a ± 5.563.26 a ± 0.10316.3 a ± 7.10875.0 a ± 26.85
Control436.25 c ± 8.422.60 c ± 0.08303.8 b ± 1.45617.3 c ± 21.75
Different letters indicate a significant difference (Tukey’s multiple range test). ± means SD.
Table 6. The impact of the application of N-fixing bacteria on yield components of faba bean in 2023.
Table 6. The impact of the application of N-fixing bacteria on yield components of faba bean in 2023.
ApplicationNumber of Seeds per PlantWeight of Seeds per Plant (g)Number of Pods per Plant
R. leguminosarum bv. viciae (inoculation)30.93 b ± 1.1815.99 b ± 0.908.53 a ± 0.39
M. symbiothicum (spraying)32.85 a ± 0.7016.79 a ± 0.448.98 a ± 0.46
Inoculation + spraying32.70 a ± 0.7517.25 a ± 0.109.18 a ± 0.28
Control26.83 c ± 1.0614.61 c ± 0.388.30 a ± 0.54
Different letters indicate a significant difference (Tukey’s multiple range test). ± means SD.
Table 7. The impact of the application of N-fixing bacteria on 1000-seed weight, seed yield, total protein content, and productivity of faba bean in 2023.
Table 7. The impact of the application of N-fixing bacteria on 1000-seed weight, seed yield, total protein content, and productivity of faba bean in 2023.
Application1000-Seed Weight (g)Seed Yield
(Mg ha−1)		Total Protein Content
(g kg−1)		Total Protein Productivity
(kg ha−1)
R. leguminosarum bv. viciae (inoculation)544.25 b ± 2.874.10 a ± 0.10291.3 c ± 1.421014.5 b ± 23.61
M. symbiothicum (spraying)556.25 a ± 4.653.93 b ± 0.02293.6 c ± 1.40980.8 c ± 6.13
Inoculation + spraying553.25 a ± 4.274.06 a ± 0.90316.4 a ± 1.251091.3 a ± 25.04
Control539.75 b ± 1.713.65 c ± 0.07306.1 b ± 1.44949.5 d ± 17.33
Different letters indicate a significant difference (Tukey’s multiple range test). ± means SD.
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Serafin-Andrzejewska, M.; Falkiewicz, A.; Wojciechowski, W.; Kozak, M. Yield and Seed Quality of Faba Bean (Vicia faba L. var. minor) as a Result of Symbiosis with Nitrogen-Fixing Bacteria. Agriculture 2025, 15, 960. https://doi.org/10.3390/agriculture15090960

AMA Style

Serafin-Andrzejewska M, Falkiewicz A, Wojciechowski W, Kozak M. Yield and Seed Quality of Faba Bean (Vicia faba L. var. minor) as a Result of Symbiosis with Nitrogen-Fixing Bacteria. Agriculture. 2025; 15(9):960. https://doi.org/10.3390/agriculture15090960

Chicago/Turabian Style

Serafin-Andrzejewska, Magdalena, Agnieszka Falkiewicz, Wiesław Wojciechowski, and Marcin Kozak. 2025. "Yield and Seed Quality of Faba Bean (Vicia faba L. var. minor) as a Result of Symbiosis with Nitrogen-Fixing Bacteria" Agriculture 15, no. 9: 960. https://doi.org/10.3390/agriculture15090960

APA Style

Serafin-Andrzejewska, M., Falkiewicz, A., Wojciechowski, W., & Kozak, M. (2025). Yield and Seed Quality of Faba Bean (Vicia faba L. var. minor) as a Result of Symbiosis with Nitrogen-Fixing Bacteria. Agriculture, 15(9), 960. https://doi.org/10.3390/agriculture15090960

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