Biomass formation and yield performance in diverse multicrops and their potential for biofuel use

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Growing agricultural plant mixtures (multicrops) contributes to increasing biomass yields, enhancing farm biodiversity, improving soil health, and promoting environmental sustainability.. However, most crop mixtures have not been studied at all. For this reason, research was conducted from 2020 to 2022 at Vytautas Magnus University, Agriculture Academy. The aim of the study was to evaluate the development and productivity of plant mixtures, as well as the related energy and environmental aspects of the applied agrotechnologies, under short-growing-season conditions. Ternary crops tended to be 14% taller, with 24% higher leaf assimilation area, 19% higher chlorophyll index, and 4–8 times higher first-year dried biomass yields than individual single-species crops. The productivity of the ternary crop reached its highest Comprehensive Evaluation Value (4.54), which was mainly influenced by the chlorophyll index of the leaves. Ternary cultivation was the most fuel-consumptive technology, with 18–32% higher fuel consumption (103.3 L ha − 1 ), due to its higher energy input; however, it generated the most significant net energy (367,668.1 MJ ha − 1 ) because of its most abundant yield of dried biomass. Ternary crop biomass pellets had the highest density (1,238 kg m -3 ), lower ash content (6%), and the highest ash shrinkage starting temperature (1042° C). It is advisable to cultivate high-capacity yielding ternary crops for one year, which have medium GHG emission and LCA impacts of the pellets produced, but the highest net energy output. Biological sciences/Ecology Earth and environmental sciences/Ecology Earth and environmental sciences/Environmental sciences Biological sciences/Plant sciences biomass biofuel pellets environment faba bean maize multicrops technical hemp Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction In recent years, the negative impacts of climate change have intensified. The warming climate exacerbates soil degradation due to intense rainfall and drought periods, reduces biodiversity and crop productivity potential (especially in southern countries), and negatively impacts economic returns [ 1 ]. In Northern countries, crop productivity potential increases due to the prolongation of the vegetative season. For example, in Lithuania, the vegetative season of crops has been extended by at least 2 – 3 weeks over the last 15 years. This allows for larger quantities of agricultural production, especially biomass. This potential is not yet fully exploited, especially in biofuel production systems, though plant biomass is one of the most important sources of renewable energy [ 2 , 3 ]. For now, the focus is on growing woody plant biomass and using forest waste. The use of plant-based biofuels significantly reduces the greenhouse effect: CO 2 emissions are close to zero, as the CO 2 released during combustion is used to produce organic matter during photosynthesis [ 4 ]. The combustion of biofuels reduces emissions of sulfur compounds to the environment, and the ashes produced can be used to fertilize the soil [ 5 ]. The development of biofuel or another environmentally friendly component production from plant biomass would contribute to the sustainability of agroecosystems, which is aimed at the circular economy [ 6 , 7 , 8 ]. Growing monocropped cultivations are still widespread in most countries to increase farm income, regardless of their negative environmental impacts [ 9 ]. Multicropped cultivations provide the opportunity to obtain a more varied multiple harvest during the vegetative season. It also has a positive effect on ecosystem services, helps to maintain and restore ecological balance through biodiversity, increases productivity and income, requires fewer inputs, reduces emissions, and increases the crop’s resilience to climatic shocks by increasing the C content of the soil [ 10 , 11 ]. Growing multicrops helps prevent the spread of weeds, diseases, and pests [ 12 ]. Growing crops together stimulates the activity of soil microorganisms, which accelerates humification processes [ 13 ]. The roots of multicrops are in different soil layers, allowing them to take up nutrients and water from different locations and thus supply them more efficiently. Growing together, plants of different species position their leaves at distinct height layers and spatial arrangements, enabling more efficient light capture, reduced mutual shading, and enhanced photosynthetic performance. [ 14 ]. Multicropping is also a promising strategy to increase land-use efficiency and cost-effectiveness of farming, especially for resource-constrained countries [ 15 ]. Technical hemp ( Cannabis sativa L.) is a promising energy crop, one of the fastest-growing plants, and accumulates the most fiber in its stems [ 16 ]. Hemp typically produces between 5.5 and 8.5 t ha -1 of dried biomass, depending on site climate, cultivar characteristics, and soil fertility [ 17 ]. Chaowana et al. [ 18 ] found that hemp stems had desirable fuel properties: high volatile matter, high calorific value, low ash content, very low nitrogen content in the emissions, and undetectable sulfur. Technical hemp can be used in biorefineries to produce bioethanol, biodiesel, biohydrogen, organic acids, and other biomaterials [ 19 ]. Maize ( Zea mays L.) is a versatile and high-yielding crop. Maize grains can be used to produce ethyl alcohol, stems and leaves can be used to produce paper, viscose, biochar, etc. Maize cultivation can produce on average 40–60 t ha -1 of green matter. Faba bean ( Vicia faba L.) is a sustainable crop that not only provides ecological services but also leaves a large amount of residual biomass after the grain harvest, which can be used for energy production [ 20 ]. A mixture of the above plants could also be a multifunctional crop, as technical hemp warrants high biomass yields and protection from pests and some diseases with low competition for sunlight. Maize ensures high crop productivity, and it has few diseases and pests in the Nordic countries. Faba beans provide ecological services to the crop by naturally providing companions with not only nitrogen but also potassium. In addition, the dried biomass residue capacity reaches 4 – 5 t ha -1 in Lithuania. What is the novelty and actuality of our research? First, ternary hemp, maize, and faba bean multifunctional crops for effective growth of abundant biomass have not yet been studied. The characteristics of their development (primarily as continuing cultivation for 3 years) in mixtures were not known. Second, the pesticide-free approach was used. Third, crops were grown for a short time, 103–105 days after germination, until the faba beans were fully mature. This ensures an early supply of plant biomass for fuel production without loss of overall productivity. Other non-bean crops can grow for longer and provide a later biomass harvest for processing. Fourth, the energy and environmental balance of multicropping technologies was assessed, and a Life Cycle Assessment (LCA) of the produced fuel pellets was performed. Fifth, the technological parameters for producing fuel pellets from a ternary crop were registered in the Patent Office of the Republic of Lithuania as an invention. The aim of the present study was to assess the development indicators and productivity of separate crop species in the tested mixtures. Additionally, this study aims to highlight the relationships between crop development and productivity indices through innovative statistical analysis tools, and to summarize the energy use and environmental aspects of the researched technologies and produced pellets. 2. Materials and methods 2.1 Site description In 2020–2022, single, binary, and ternary crops of maize ( Zea mays L.), technical hemp ( Cannabis sativa L.), and faba bean ( Vicia faba L.) were sown at the Experimental Station of Vytautas Magnus University Agriculture Academy (VMU AA) at the following coordinates: 54°53′7.5″ N 23°50′18.11″ E. The Experimental Station is located on the south-western side of Kaunas city, on the left bank of the Nemunas River, in the central part of Lithuania (Supplementary Fig. 1). The soil at the experimental site is a silty loam (46% sand, 42% silt, 12% clay) Endohypogleyic-Eutric Planosol (Ple-gln-w) [21], with a texture of silty light loam on heavy loam. A soil pH KC was 7.3–7.8, total nitrogen content – 0.08–0.13%, organic matter content – 2.6–2.9%, available potassium – 118 mg·kg -1 , available phosphorus – 189–280 mg·kg -1 , available sulfur – 1.2–2.6 mg·kg -1 , available magnesium – 436–790 mg·kg -1 . Lithuania has sufficient precipitation in all seasons, especially in the warmer months, but recently, droughts have become more frequent in the summer. The average annual precipitation in Lithuania is 600–900 mm, and evaporation is around 500 mm. The annual temperature is around 6.5–7.5° C. During the experiment, meteorological conditions varied considerably each year. The air temperature in 2020 was close to the long-term average, with only July being slightly warmer. In that year, May and June had higher precipitation, but in April and July, which are particularly important for plant development and productivity, the crops experienced a lack of moisture (Supplementary Fig. 2). 2021 was a critical year for crop growth, with above-average temperatures and very low precipitation in June and July. In contrast, August was cooler compared to the other years of study and the long-term average, but with sufficient precipitation. In 2022, air temperature was lower than usual throughout the vegetation period, except at the end of the vegetation period, when the temperature in August was higher than the long-term average and there was a severe lack of humidity. 2.2. Experimental design and agronomic operations Before setting up the experiment, oats ( Avena sativa L.) were grown in the experimental site. The experiment consisted of 7 treatments, 21 experimental plots in total, arranged with three replications in a randomized complete block design (RCBD). The size of the plot was 8 m 2 . Table 1 shows the list of performed treatments and their abbreviations. Maize ( Zea mays L.) (cultivar “Pioneer” hybrid P7034), technical hemp ( Cannabis sativa L.) (cultivar “Austa SK”), and faba bean ( Vicia faba L.) (cultivar “Vertigo”) as single, binary, and ternary crops, respectively, were grown to obtain the highest possible biomass yield at low-input pesticide-free conditions and by continuing the growth of crops. The soil of the experiment was plowed with a Gamega PP-3-43 plow (Lithuanian producer) with semi-helical shellboards in each fall (usually in October) and cultivated with a complex Laumetris KLG-3.6 cultivator (Lithuanian producer) in spring (usually in April) before sowing. Before sowing, mineral complex fertilizer NPK 15:15:15 (300 kg·ha −1 ) was treated by hand equipment. This fertilization rate is insufficient for achieving high biomass yields; however, we aim to demonstrate a more pronounced effect of continued multicropping on soil fertility. We expected that the ecological services of faba beans would compensate for the negative impact of crops continuing to grow on the soil properties. Experimental plots were sown by hand equipment according to specially created seed distributing schemes [22] (Supplementary Fig. 3). The presented plant sowing schemes ensured the development of crop mixtures with minimal competition (Supplementary Fig. 4). Sowing rates depend on the scheme and are presented in more detail in Supplementary Table 1. The crops’ inter-rows were loosened 1–2 times until the crops covered the inter-rows. Crop biomass production was harvested after a short 103–105-day vegetative season, when the faba beans reached full maturity (Supplementary Table 2). At that time, the biomass of maize and hemp had not yet reached its maximum. Still, faba beans produce abundant and valuable biomass (especially in the first year of growing), which we did not want to lose because in August, there is usually a shortage of biomass for fuel production in Lithuania. The applied technological operations and technical means, and their energy and CO 2eq. balance are discussed in more detail in our previous article [23]. 2.3. Methods and analysis To determine crop development indicators, measurements were made during the vegetation period when faba bean began to bloom (maize BBCH 51–53, technical hemp BBCH 60–62, faba bean BBCH 63–65), assessing plant height, leaf chlorophyll index, assimilation area, and green (fresh) biomass of each crop species. At the end of the plant vegetation period (maize BBCH 79–80, technical hemp BBCH 81–83, faba bean BBCH 86–88), not only was the green biomass of maize, hemp, and faba bean assessed separately, but also the total green and dried biomass of the crops per unit area. For development indicator studies, five plants of each crop species were cut out in each experimental plot. The assimilation area of plant leaves (cm 2 ) was determined with a leaf area meter Win Dias (“Delta-T Devices” Ltd, UK). The chlorophyll index of plant leaves was measured with a chlorophyll meter CCM–200 Plus. The height of the plants was measured, and they were weighed, thus determining their green biomass. At the end of the vegetation period, samples were taken at least five spots per plot, in a 0.5 m longitudinal row. An average sample was formed. A total of 36 research samples were formed. To determine the dried biomass, the samples were dried in a thermostat at a temperature of 105 °C. The plant biomass grown can be used in energy and other industries, especially in the production of biofuels, fertilizers, bio-additives, etc.; therefore, its elemental composition is important (Supplementary Table 3). In Supplementary Table 4, the main biofuel pellet characteristics are presented. The chemical composition was tested in the laboratories of the Lithuanian Research Centre of Agriculture and Forestry in Kaunas, Lithuania. The experimental data were analyzed using one-way ANOVA, and the treatment effect was estimated by the F-test and the least significant difference (LSD). SYSTAT and SELEKCIJA software were employed. Significant differences between the treatments are marked with different lowercase letters, when P≤0.05>0.01 . Significant differences between the vegetation conditions of individual years of the experimentation are marked with different uppercase letters, when P≤0.05>0.01 . A correlation analysis was performed with SigmaStat software. Principal Component Analysis (PCA) was used to establish the correlation between different indicators in single, binary, and ternary crops. This analysis creates new artificial variables (principal components) based on the analysed variables. Its central premise was the ability to visualise the relationships between individual variables in a two-dimensional diagram showing the coordinate system of the first two principal components. Based on the position of the vectors in space, it is possible to determine which variables are correlated with each other. A lower angle between the vectors shows the stronger positive correlation. When the vectors are aligned on the same line but in opposite directions, there is a strong negative correlation between the variables. However, when the vectors are aligned at an angle close to 90 degrees, there is no correlation [24]. The results of the investigations were also grouped using cluster analysis. The clustering of all tested single, binary, and ternary crops, considering the estimated average height of crops, leaf assimilation area, and chlorophyll index, crop green (fresh) and dried biomass, was carried out according to the Ward criterion using Euclidean distance matrices. Statistical analysis was performed using the Statistica software package Statistica 10 (TIBCO Software Inc., Palo Alto, CA, USA). Calculations were performed at Vytautas Magnus University Agriculture Academy. 3. Results and discussion 3.1. Average plant height The intensity of crop diversification (factor A) negatively affected the average heights of maize and technical hemp. They were significantly higher when grown as single crops, and the lowest when grown as a ternary crop. Maize and technical hemp were also taller when grown together with faba bean (Table 2). Fisher et al. [25] in Germany and Hirpa [26] in Ethiopia found no effect on maize height when grown in a binary crop with faba bean. Li et al. [27] found the negative effect. The height indicators of faba bean differed from those of maize and technical hemp. The lowest faba bean height was observed in a single crop, with a significant difference of 10.9 cm when grown together with maize. Regarding the effect of crop continuing growing (factor B) on plant height, it was found that the lowest height of maize and technical hemp was in the last year of the experiment, i.e., the heights were 1.6–1.9 times less because of degradation of most soil properties [28]. A comprehensive assessment of the different indicators studied showed that plant height in a hemp crop was the highest expressed at crop development and productivity level compared to other experimental treatments [29] (Supplementary Fig. 5). 3.2. Leaf assimilation area The increase in the number of plant species in the crop influenced the decrease in the assimilation area of their leaves. The largest leaf area of maize and technical hemp was determined when they grew as single crops (Table 3). When maize grew together with faba bean, their leaf assimilation area was 16% higher than in other diversified crops. This is likely because faba beans offer less competition for maize growth and supplement the soil with nitrogen, a nutrient essential for plant growth. Similarly, Li et al. [30] and Yang et al. [31] also found that the maize-soybean binary crop increased the assimilation area of maize leaves. Yang et al. [32] and Corre-Hellou et al. [33] confirmed these results with pea ( Pisum sativum L.) cover- and inter-crops. A comprehensive data analysis revealed that maize leaves’ assimilation area was the highest at crop development and productivity levels compared to other experimental treatments (Supplementary Fig. 5). Additionally, we found a positive moderate relationship between maize leaf assimilation area and plant height (r = 0.64, P ≤ 0.050 > 0.010). The largest leaf assimilation area of hemp was found in the binary crop with faba bean, and the highest area of faba bean leaves was found in the binary crop with maize. On the contrary, Wu et al. [34] found that when faba bean grows together with maize, increased competition for sunlight causes faba bean stems to lengthen and reduces their leaf assimilation area. A significant effect of vegetative conditions on the assimilation area of maize and technical hemp leaves was found. In the second year of the study, the assimilation area of maize and technical hemp leaves was significantly higher. Contrary to expectations, the hot and dry months of June-July in that year did not harm the development of these crops. Different vegetation conditions in individual years of the study influenced a consistent increase in the assimilation area of faba bean leaves. 3.3. Chlorophyll index of plant leaves The chlorophyll index of leaves of each single crop was usually higher than that of diversified crops (Table 4). Similarly, Peñafiel–Sandova [35] also found the highest chlorophyll index in the maize single crop. According to the average data of our experiment, factor B, the chlorophyll index in the last year of the experiment decreased 3 times in maize and 1.4 times in technical hemp crops compared to the first year of the study. The opposite results were obtained in faba bean crops, since the chlorophyll index was significantly highest in the third year of the study. According to the comprehensive analysis of the research data, the crop leaf chlorophyll index was the most common in the tested ternary crop. In another cultivation, the rating of this index mainly did not exceed the evaluation threshold (Supplementary Fig. 5). 3.4. Green biomass of individual plant species An interaction was found between the effect of crop diversification and vegetation conditions of individual research years on the average green biomass of plants at the end of the vegetation. Crop diversification negatively affected the green biomass of individual plants, since it was highest in single crops of maize, technical hemp, and faba bean (Tables 5 and 6). Among the diversified crops, the highest green biomass of maize and technical hemp was found when they grew together with faba bean. Here, the green biomass of maize was 3 times higher, and that of technical hemp was 2 times higher than in the ternary crop. Combining maize with various legumes, such as faba bean, improved maize crop growth, increased its green biomass, and increased plant quality [36]. The green biomass of faba bean did not differ significantly in the diversified crops. The vegetation conditions of the research years also had a significant impact on the green biomass of maize, technical hemp, and faba bean. The highest green biomass of these plants was in the first year of the experiment, and then it consistently decreased. In the third year of the experiment, the green biomass of maize, hemp, and faba bean decreased by 2–4 times. Maize green biomass was partly dependent on its leaf chlorophyll index (r = 0.58, P ≤ 0.050 > 0.010) and leaf assimilation area (r = 0.72, P ≤ 0.050 > 0.010). Technical hemp dependencies were similar (r = 0.87, r = 0.86, P ≤ 0.010 > 0.001). A statistically significant correlation was also established between faba bean leaf assimilation area and capacity of green biomass (r = 0.87, P ≤ 0.010 > 0.001). Plants’ green biomass also partly depended on the crop density and average height of plants. 3.5. Total biomass of cultivations An interaction was established between the effect of crop diversification and the vegetative conditions of individual research years on the average green biomass of crops at the end of the plant vegetation. Among single crops, the highest green biomass per unit area was observed for maize (4,410.7 g m -2 ). Among diversified crops, the highest green biomass was obtained when growing a binary crop of maize and faba bean (3,421.5 g m -2 ). Shtaya et al. [37] also confirmed that faba bean, when mixed with other crops, increased the total green biomass of the crop. Streit et al. [38] obtained the opposite results and claim that faba bean in mixtures produces an average of 5% more green biomass than in single crops. In our experiment, the green biomass of the ternary crop was 25% lower than that of the binary crop of maize and faba bean (Table 7). The vegetative conditions of individual research years also had a significant effect on the green biomass of crops at the end of the plant vegetation period. It was significantly highest in the first year of the experiment and decreased by 30% in the second and third research years. As expected, the ternary crop had the highest total dried biomass, which was 8 times higher than that of the single maize crop. This is because at the time of biomass harvesting, maize contained a large amount of water, while the faba bean biomass was close to air-dried (BBCH 95–97). According to Ciampitti et al. [39], in faba bean, the highest N 2 fixation occurs at the flowering stage. Therefore, harvesting at the right time is very important not only to ensure the best crop yield and the highest possible N 2 contribution to subsequent crops, but also to maximize biomass energy yield. This was observed in our experiment, where maize benefited from the ecological services provided by faba bean. Faba beans, together with companions, can also increase not only total yield and income, but reduce crop weediness and disease, increase land use efficiency, and thus increase crop sustainability [37, 40]. Moreover, the dried biomass of all single crops was lower compared to the biomass of diversified crops (Table 8). Dzvene [41] obtained similar results. The crops’ continued growth harmed their total biomass yields. Only in the first year of the study did we receive the highest yields (especially in the ternary crop), and in the second and last years of the study, they decreased by about 6.5 times due to the degradation of soil properties. Bybee–Finley et al. [42] found that in an experiment with millet, sorghum, and hemp, in the first year of crop cultivation, the biomass of plants grown in mixtures was higher than that of single crops. After conducting a comprehensive assessment of the experimental data, it was found that the ternary crop had the highest CEI value; therefore, it was the most effective in growing plant total biomass. The formation of higher dried biomass in the ternary crop was most influenced by the higher leaf chlorophyll index (Supplementary Fig. 5, marked in yellow). A higher CEI value was also obtained in the binary M+H cultivation. This confirms the statement of Branca et al. [43] and Hu et al. [44] that maize and technical hemp are the most promising crops in the field of biomass processing. A generalized cluster analysis of our experimental data showed that maize and hemp were more efficiently grown in intercropping with faba beans due to their higher biomass capacity (Fig. 1). The PCA was also done for different species of companion crops in cultivation: Maize. In 2020, the aboveground biomass of maize in the middle and end of the vegetative season was closely related to plant height and plant photosynthetic indicators ˗ plant leaf assimilation area and chlorophyll index in leaves (Fig. 2). In 2021, the aboveground biomass of maize in the middle of the vegetation correlated with plant leaf assimilation area and chlorophyll index in leaves. The latter indicators had less influence on the aboveground biomass of plants at the end of the vegetation. In 2022, similar trends were found as in 2021. Technical hemp. In 2020, the aboveground biomass of hemp in the middle and end of the vegetative period was most influenced by plant average height. The influence of leaf assimilation area was lower (Fig. 3). In 2021, two groups of closely correlated indicators emerged: the first group consisted of the aboveground biomass of plants at the beginning of the vegetative period and the leaf assimilation area, and the second group consisted of the aboveground biomass of plants at the end of the vegetative period and plant height. In 2022, the aboveground biomass of hemp in the middle of the vegetative season was closely related to the plant leaf assimilation area. The aboveground biomass at the end of the vegetative period was more dependent on the plant height. Faba bean. In 2020, the aboveground biomass of faba beans in the middle and end of the vegetative period was closely related to plant photosynthetic indicators – plant leaf assimilation area and chlorophyll index (Fig. 4). In 2021, the aboveground biomass of faba beans at the middle of the vegetative period was less influenced by plant height and photosynthetic indicators. In 2022, the aboveground biomass of faba beans at the end of the vegetative period depended most on the chlorophyll concentration in the leaves. In summary, it can be stated that crop diversification had a positive impact on the total yield of dried biomass. The dried biomass of the ternary crop was 4–8 times higher compared to single crops, and 2 times higher than that of binary crops. It is recommended to grow low-fertilized and pesticide-free ternary crops in existing on-farm crop rotations for one year only. Continuing crops need to be well fertilized, because biomass productivity gradually decreases. Aubin et al. [45] concluded that higher N rates can maximally improve hemp growth, plant height, and biomass. 3.6. Technological, energy, and environmental aspects All tested cropping technologies include stubble cultivation (depth 12–15 cm) in fall, deep plowing before wintering, pre-sowing cultivation, and fertilization. One-pass conventional sowing was used for single crops, while two-pass ternary sowing was used for binary and ternary crops. Inter-row loosening (2–3 cm depth) was performed twice for all cultivations, except the ternary crop. In the ternary crop, the inter-row loosening was performed once due to higher crop densities. In calculations, we simulated one-pass biomass harvesting (low harvester load) for M, FB, and M+FB crops, one-pass biomass harvesting (high harvester load) for H, M+H, and H+FB crops, and two-pass biomass harvesting (high harvester load) for the ternary crop. Tractor power (kW) for calculations ranged from 45–67 kW (for sowing, fertilization, and shallow loosening) to 102 kW (for deeper tillage operations). The combine harvester power remained constant at 250 kW. We adjusted the field capacity from 1.82 to 0.68 ha/harvester load. Technological aspects are discussed in more detail in Romaneckas et al. [23]. The highest consumption of fuel was calculated for the ternary crop (M+H+FB) (103.3 L ha −1 ) due to a higher number and more powerful operations (Supplementary Table 5). Maize and faba bean single cropping used the lowest amount of fuel. Although the technology for growing the ternary crop required more energy input, the yield obtained compensated for this. So, the highest net energy (367,668.1 MJ ha −1 ) was also obtained in the ternary crop. Obviously, growing more plant companions in the same crop results in higher net energy [46]. The lowest total CO 2eq. (greenhouse gases) emissions were calculated for single hemp and faba bean cultivations, and the highest were for M+H and M+FB binary crops. GHG emission of ternary crop was average (1,541.90 kg ha −1 CO2 eq. ) [23]. The short-growing biomass of the tested maize-hemp-faba bean ternary crop can be effectively used for energy purposes and crop fertilization, because the pH is close to neutral, contains about 1% of nitrogen and potassium, and about 0.2% of phosphorus (Supplementary Table 2), and the average yield of total dried biomass reaches almost 20 t ha -1 . A National invention patent had been registered to produce pellets from the biomass of the ternary crop discussed [47]. These pellets had a higher density than those made from biomass of other cultivations, one of the lowest ash contents, and the highest ash shrinkage starting temperature. They met the requirements of the ISO 17225-6:2021 standard (Supplementary Table 3). Moreover, the burning of pellets in household low-power boilers did not have negative environmental consequences [48]. Unlike the evaluation of crop production technologies, the Life Cycle Assessment (LCA) of the impact of pellet disposition and use (transportation, heat production, and ash utilization) on abiotic depletion, global warming potential, acidification, and eutrophication indices showed that binary M+H crops had the lowest impact on the environment. Pellets made from ternary crop biomass had an average dimension [49]. 4. Conclusions Due to the eco-service of faba beans, maize and technical hemp mixtures with faba beans tended to have 14% greater height, 24% higher leaf assimilation area, and 19% higher chlorophyll index. With the increase in the number of plant species in the crop mixtures, the biomass productivity of each plant species decreased, but the total biomass productivity per plot area increased. In the first year of the experiment, a ternary maize–technical hemp–faba bean crop produced 4–8 times more dried biomass than individual single species. Multicropping continuation for the next two years decreased biomass yields by up to 11 times. Ternary multicrop productivity reached the highest CEI value (4.54), which was mainly influenced by the chlorophyll index of the plant leaves. Ternary cultivation was the most fuel-consuming technology (18–32% higher fuel consumption) – 103.3 L ha − 1 due to higher energy input. However, it could be characterized by the most significant net energy (367,668.1 MJ ha − 1 ) because of the most abundant yield of dried biomass. The pellets produced from ternary crop biomass met the standards, had the highest density (1,238 kg m -3 ), one of the lowest ash contents (6%), and the highest ash shrinkage starting temperature (1042°C). The LCA of pellet disposition and use showed that the binary M + H crop had the lowest impact on the environment. However, biomass growing technology was one of the most contaminating; therefore, it is advisable to develop high-capacity yielding ternary crops, whose GHG emissions and LCA effects on the produced pellets had an average dimension. Contributions Conceptualization, K.R., J.B.; methodology, K.R., J.B.; software, J.B.; validation, J.B., K.R.; formal analysis, J.B., K.R., A.M.; investigation, J.B., R.K., A.S. and K.R.; resources, J.B, R.K., A.S. and K.R.; data curation, J.B.; writing—original draft preparation, J.B., K.R.; writing—review and editing, K.R., J.B., R.K. and A.S; visualization, J.B., K.R., A.M.; supervision, K.R. All authors have read and agreed to the published version of the manuscript. Declarations Contributions: Conceptualization, K.R., J.B.; methodology, K.R., J.B.; software, J.B.; validation, J.B., K.R.; formal analysis, J.B., K.R., A.M.; investigation, J.B., R.K., A.S. and K.R.; resources, J.B, R.K., A.S. and K.R.; data curation, J.B.; writing—original draft preparation, J.B., K.R.; writing—review and editing, K.R., J.B., R.K. and A.S; visualization, J.B., K.R., A.M.; supervision, K.R. All authors have read and agreed to the published version of the manuscript. Conflict of Interest: The authors declare that they have no known competing financial interests or personal relationships that may influence the work reported in this paper. Funding: The reported work in this article was partially supported by the Ministry of Education, Science and Sports of the Republic of Lithuania and Research Council of Lithuania (LMTLT) under the Program ‘University Excellence Initiative’ Project ‘Development of the Bioeconomy Research Center of Excellence’ (BioTEC), agreement No S-A-UEI-23-14. Availability of data and materials: described data and materials are available from the corresponding author upon request. The methodology for the comprehensive assessment and a detailed calculation example is provided in our previous article Kimbirauskienė R, Sinkevičienė A, Švereikaitė A, Romaneckas K. The Complex Effect of Different Tillage Systems on the Faba Bean Agroecosystem. Plants. 2024; 13(4):513. https://doi.org/10.3390/plants13040513, Supplementary materials, Table S1. The meteorological indicators presented in the article were obtained from the Lithuanian Hydrometeorological Service, Kaunas Meteorological Station. https://www.meteo.lt/en/organization/structure-and-contacts/contacts/" in Supplementary materials, Figure 2. Data on main characteristics of grown multi crop solid fuel pellets was referred from Petlickaitė, R., Jasinskas, A., Domeika, R., Pedišius, N., Lemanas, E., Praspaliauskas, M., Kukharets, S. Evaluation of the Processing of Multi-Crop Plants into Pelletized Biofuel and Its Use for Energy Conversion, Processes. 11(2), 421, https://doi.org/10.3390/pr11020421 (2023) in Supplementary materials, Table 3. References Oishy, M. N. et al. A. Unravelling the effects of climate change on the soil-plant-atmosphere interactions: A critical review. SEH. 10130, (2025). https://doi.org/10.1016/j.seh.2025.100130 Erdiwansyah, A. G. et al. Prospects for renewable energy sources from biomass waste in Indonesia. Case Stud. Chem. Environ. Eng. 10 , 100880. https://doi.org/10.1016/j.cscee.2024.100880 (2024). Roy, R. et al. Steam explosion treated biomass as a renewable fuel source: A review from collection to combustion. Fuel 378 , 132883. https://doi.org/10.1016/j.fuel.2024.132883 (2024). Licata, M. et al. A comparative study to assess the production of two oilseed crops (Brassica carinata A. Braun and Carthamus tinctorius L.) and the energy potential of their agricultural biomass residues. Heliyon 10 (22), e38654. https://doi.org/10.1016/j.heliyon.2024.e38654 (2024). Jasinskas, A., Petlickaitė, R., Praspaliauskas, M., Romaneckas, K. & Sinkevičienė, A. Impact of ash obtained after multi-crop pellet burning on spring barley fertilization. In:Engineering for Rural Development: 23rd International Scientific Conference, ERDev 2024-Proceedings. Latvia University of Life Sciences and Technologies, 23, pp. 644–649 (2024). Muthulakshmi, C., Sivaranjani, R. & Selvi, S. Modification of sesame (Sesamum indicum L.) for Triacylglycerol accumulation in plant biomass for biofuel applications. Biotechnol. Rep. 32 , e00668. https://doi.org/10.1016/j.btre.2021.e00668 (2021). Wang, T. et al. From intercropping to monocropping: The effects of Pseudomonas strain to facilitate nutrient efficiency in peanut and soil. Plant. Physiol. Biochem. 219 , 109378. https://doi.org/10.1016/j.plaphy.2024.109378 (2025). Kumar, R. K. S., Sasikumar, R. & Dhilipkumar, T. Exploiting agro-waste for cleaner production: A review focusing on biofuel generation, bio-composite production, and environmental considerations. J. Clean. Prod. 435 , 140536. https://doi.org/10.1016/j.jclepro.2023.140536 (2024). Alcon, F. et al. Cost benefit analysis of diversified farming systems across Europe: Incorporating non-market benefits of ecosystem services. Sci. Total Environ. 912 , 169272. https://doi.org/10.1016/j.scitotenv.2023.169272 (2024). Hashakimana, L., Tessema, T., Niyitanga, F., Cyamweshi, A. R. & Mukuralinda, A. Comparative analysis of monocropping and mixed cropping systems on selected soil properties, soil organic carbon stocks, and simulated maize yields in drought-hotspot regions of Rwanda. Heliyon 9 , e19041. https://doi.org/10.1016/j.heliyon.2023.e19041 (2023). Léonidas, H. et al. Monocropping vs mixed cropping systems under a changing climate: Smallholder farmers' perceptions and farm profitability in Eastern Rwanda. Environ Sustain Ind . 24 , 100527. https://doi.org/10.1016/j.indic.2024.100527 (2024). Su, B., Liu, X., Cui, L., Xiang, B. & Yang, W. Suppression of Weeds and Increases in Food Production in Higher Crop Diversity Planting Arrangements: A Case Study of Relay Intercropping. Crop Sci. 58 (4), 1729–1739. https://doi.org/10.2135/cropsci2017.11.0670 (2018). Tariq, A. et al. Combining different species in restoration is not always the right decision: Monocultures can provide higher ecological functions than intercropping in a desert ecosystem. J. Environ. Manag . 357 , 120807. https://doi.org/10.1016/j.jenvman.2024.120807 (2024). Chimonyo, V. G. P. et al. Chapter 18 - Yield and water use gaps in cereal multicrop systems in sub-Saharan Africa under climate change. Food Processing Technology. Principles and Practice. A volume in Woodhead Publishing Series in Food Science, Technology and Nutrition, pp. 313–329 (2021). Vanino, S. et al. Rarina, R. A comprehensive assessment of diversified cropping systems on agro-environmental sustainability in three Mediterranean long-term field experiments. Eur. J. Agron. 140 , 126598. https://doi.org/10.1016/j.eja.2022.126598 (2022). Mamimin, C., O-Thong, S. & Reungsang, A. Enhancing biogas production from hemp biomass residue through hydrothermal pretreatment and co-digestion with cow manure: Insights into methane yield, microbial communities, and metabolic pathways. J. Environ. Manag . 370 , 123039. https://doi.org/10.1016/j.jenvman.2024.123039 (2024). Vávrová, K. et al. Economic evaluation of Hemp’s (Cannabis sativa) residual biomass for production of direct energy or biochar. Fuel 329 , 125435. https://doi.org/10.1016/j.fuel.2022.125435 (2022). Chaowana, P. et al. Utilization of hemp stalk as a potential resource for bioenergy. Mat. Sci. En Tech. 7 , 19–28. https://doi.org/10.1016/j.mset.2023.07.001 (2024). Tripathi, M. et al. Conversion technologies for valorization of hemp lignocellulosic biomass for potential biorefinery applications. 320 , 124018, (2023). https://doi.org/10.1016/j.seppur.2023.124018 Gómez, L. D. et al. Caruso, L. Valorising faba bean residual biomass: Effect of farming system and planting time on the potential for biofuel production. Biomass Bioenerg . 107 , 227–232. https://doi.org/10.1016/j.biombioe.2017.10.019 (2017). IUSS Working Group WRB. World Reference Base for Soil Resources: International soil classification system for naming soils and creating legends for soil maps, 4th ed., Vienna, Austria, (2022). Romaneckas, K. et al. Short-Term Impact of Multi-Cropping on Some Soil Physical Properties and Respiration. Agron . 12, 141, (2022). https://doi.org/10.3390/agronomy12010141 Romaneckas, K., Švereikaitė, A., Kimbirauskienė, R., Sinkevičienė, A. & Balandaitė, J. The Energy and Environmental Evaluation of Maize, Hemp and Faba Bean Multi-Crops. Agron 13 , 2316. https://doi.org/10.3390/agronomy13092316 (2023). Jolliffe, I. T. & Cadima, J. Principal component analysis: a review and recent developments. Philosophical Transactions of the Royal Society. Math. Phys. Eng. Sci. 374 , 1–16. https://doi.org/10.1098/rsta.2015.0202 (2016). Fisher, J., Bohm, H. & Heβ, J. Maize–bean intercropping yields in Northern Germany are comparable to those of pure silage maize. Eur. J. Agron. 112 , 125–147. https://doi.org/10.1016/j.eja.2019.125947 (2020). Hirpa, T. Effect of intercrop row arrangement on maize and haricot bean productivity and the residual soil. GJSFR 14 , 1–9 (2014). Li, Z. et al. The synergistic priming effect of exogenous salicylic acid and H2O2 on chilling tolerance enhancement during maize ( Zea mays L.) seed germination. Front. Plant. Sci. 8 , 1153. 10.3389/fpls.2017.01153 (2017). Balandaitė, J. Crop biodiversity influence on multi-crops sustainability, productivity and energy efficiency. Doctoral Thesis. Vytautas Magnus University, Kaunas, Lithuania, (2023). Balandaitė, J., Romaneckas, K., Kimbirauskienė, R. & Sinkevičienė, A. Comprehensive Assessment of the Effect of Multi-Cropping on Agroecosystems. Plants 13 (10), 1372. https://doi.org/10.3390/plants13101372 (2023). Li, Y. et al. Maize–soybean relay cropping increases soybean yield synergistically by extending the post-anthesis leaf stay-green period and accelerating grain filling. Crop J. 11 (6), 1921–1930. https://doi.org/10.1016/j.cj.2023.05.011 (2023). Yang, Z. et al. GmPTF1 modifies root architecture responses to phosphate starvation primarily through regulating GmEXPB2 expression in soybean. Plant. J. 107 , 525–543. https://doi.org/10.1111/tpj.15307 (2021). Yang, H. et al. Yield photosynthesis and leaf anatomy of maize in inter- and mono-cropping systems at varying plant densities. Crop J. 10 , 893–903 (2022). Corre-Hellou, G., Fustec, J. & Crozat, Y. Interspecific competition for soil N and its interaction with N2 fixation, leaf expansion and crop growth in pea–barley intercrops. Plant. Soil. 282 , 95–208 (2006). Wu, Y. et al. Combining modelling and experiment to quantify light interception and inter row variability on intercropped soybean in strip intercropping. Eur. J. Agron. 164 , 127508. https://doi.org/10.1016/j.eja.2025.127508 (2025). Peñafiel–Sandova, Z. B. Effect of urea on lead absorption in corn ( Zea mays L), spinach ( Spinacia olerácea L.) and cabbage ( Brassica olerácea L). Agron. Colomb . 38 (2), 205–217. 10.15446/agron.colomb.v38n2.85082 (2021). Stoltz, E. & Nadeau, E. Effects of intercropping on yield, weed incidence, forage quality and soil residual N in organically grown forage maize ( Zea mays L.) and faba bean ( Vicia faba L). Field Crops Res. 169 , 21–29. https://doi.org/10.1016/j.fcr.2014.09.004 (2014). Shtaya, M. J. Y. et al. Effects of crop mixtures on rust development on faba bean grown in Mediterranean climates. Crop Prot. 146 , 105686. https://doi.org/10.1016/j.cropro.2021.105686 (2021). Streit, J., Meinen, C., Nelson, W. C. D. & Rauber, R. Above and belowground biomass in a mixed cropping system with eight novel winter faba bean genotypes and winter wheat using FTIR spectroscopy for root species discrimination. Plant. Soil. 36 , 141–158. https://doi.org/10.1007/s11104-018-03904-y (2019). Ciampitti, I. A. et al. M. Revisiting Biological Nitrogen Fixation Dynamics in Soybeans. Front. Plant. Sci. 12 , 727021. https://doi.org/10.3389/fpls.2021.727021 (2021). Nurgi, N., Tana, T., Dechassa, N., Tesso, B. & Alemayehu, Y. Effect of spatial arrangement of faba bean variety intercropping with maize on yield and yield components of the crops. Heliyon 9 (6), e16751. https://doi.org/10.1016/j.heliyon.2023.e16751 (2023). Dzvene, A. R., Tesfuhunei, W. A., Walker, S. & Ceronio, G. Optimizing the planting time and stand density of sunn hemp intercropping for biomass productivity and competitiveness in a maize-based system. Field Crops Res . 304 , 109179. https://doi.org/10.1016/j.fcr.2023.109179 (2023). Bybee-Finley, K. A., Mirsky, S. B. & Ryan, M. R. Crop Biomass Not Species Richness Drives Weed Suppression in Warm-Season Annual Grass–Legume Intercrops in the Northeast. Weed Sci. 65 , 669–680 (2017). https://www.jstor.org/stable/26420910 Branca, C., Blasi, C. D. & Galgano, A. Experimental analysis about the exploitation of industrial hemp (Cannabis sativa) in pyrolysis, Fuel Process. Technol . 162, 20–29, (2017). https://doi.org/10.1016/j.fuproc.2017.03.028 Hu, E. et al. Pyrolysis behaviors of corn stover in new two-stage rotary kiln with baffle. J. Anal. Appl. Pyrol. 161 , 105398. https://doi.org/10.1016/j.jaap.2021.105398 (2022). Aubin, M. P. et al. Industrial hemp response to nitrogen, phosphorus, and potassium fertilization. Crop Forage Turf Man. 1 (1), 1–10. https://doi.org/10.2134/cftm2015.0159 (2016). Šarauskis, E. et al. Energy balance, costs and CO2 analysis of tillage technologies in maize cultivation. Energy 69 , 227–235. https://doi.org/10.1016/j.energy.2014.02.090 (2014). Jasinskas, A. et al. Hemp and Bean Multi-Crop Biomass Pellets and/or Fertilizer. Patent LT6998B. Available online: (2022). https://search.vpb.lt/pdb/patent/dossier/48325/text Petlickaitė, R. et al. Evaluation of the Processing of Multi-Crop Plants into Pelletized Biofuel and Its Use for Energy Conversion, Processes . 11(2), 421, (2023). https://doi.org/10.3390/pr11020421 Petlickaitė, R. et al. Evaluation of Multi-Crop Biofuel Pellet Properties and the Life Cycle Assessment. Agric 14 , 1162. https://doi.org/10.3390/agriculture14071162 (2024). Tables Table 1 to 8 are available in the Supplementary Files section. Additional Declarations No competing interests reported. Supplementary Files Tables1121.docx Supplementarymaterials1121.docx Cite Share Download PDF Status: Published Journal Publication published 30 Mar, 2026 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 06 Mar, 2026 Reviews received at journal 06 Mar, 2026 Reviewers agreed at journal 22 Feb, 2026 Reviewers agreed at journal 21 Feb, 2026 Reviewers agreed at journal 18 Jan, 2026 Reviews received at journal 11 Jan, 2026 Reviewers agreed at journal 05 Jan, 2026 Reviewers invited by journal 02 Jan, 2026 Editor assigned by journal 26 Nov, 2025 Submission checks completed at journal 26 Nov, 2025 First submitted to journal 26 Nov, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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2","display":"","copyAsset":false,"role":"figure","size":70493,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figures11212.png","url":"https://assets-eu.researchsquare.com/files/rs-8209611/v1/bd680ea65b9c7ff179cf16ae.png"},{"id":96933168,"identity":"dda8f687-f9d3-4ec4-9a79-10d825004463","added_by":"auto","created_at":"2025-11-27 15:50:16","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":61028,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figures11213.png","url":"https://assets-eu.researchsquare.com/files/rs-8209611/v1/e09fe82b482e1f9bc48b07e5.png"},{"id":97136399,"identity":"6ca9bf84-64aa-4bb6-b2e2-3acff59e6e4f","added_by":"auto","created_at":"2025-12-01 09:56:30","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":62778,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figures11214.png","url":"https://assets-eu.researchsquare.com/files/rs-8209611/v1/a18c1b17c33f6a49e9704d33.png"},{"id":106343385,"identity":"d99d6bd0-112d-4a10-b8d0-9429bb7ec289","added_by":"auto","created_at":"2026-04-07 16:04:23","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":926557,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8209611/v1/ee578e4a-2157-4fbb-8ea2-bcda192600a1.pdf"},{"id":97135638,"identity":"15145660-cae1-4150-833f-469b97279069","added_by":"auto","created_at":"2025-12-01 09:52:12","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":37415,"visible":true,"origin":"","legend":"","description":"","filename":"Tables1121.docx","url":"https://assets-eu.researchsquare.com/files/rs-8209611/v1/d19b3fe2921fc68f7c3f6ba9.docx"},{"id":96933133,"identity":"57f6175c-35c3-4fd2-8e3c-0ef9d6d8214a","added_by":"auto","created_at":"2025-11-27 15:49:20","extension":"docx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":2750938,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarymaterials1121.docx","url":"https://assets-eu.researchsquare.com/files/rs-8209611/v1/44516a9421702d23b315588e.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Biomass formation and yield performance in diverse multicrops and their potential for biofuel use","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eIn recent years, the negative impacts of climate change have intensified. The warming climate exacerbates soil degradation due to intense rainfall and drought periods, reduces biodiversity and crop productivity potential (especially in southern countries), and negatively impacts economic returns [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. In Northern countries, crop productivity potential increases due to the prolongation of the vegetative season. For example, in Lithuania, the vegetative season of crops has been extended by at least 2\u003cb\u003e\u0026ndash;\u003c/b\u003e3 weeks over the last 15 years. This allows for larger quantities of agricultural production, especially biomass. This potential is not yet fully exploited, especially in biofuel production systems, though plant biomass is one of the most important sources of renewable energy [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. For now, the focus is on growing woody plant biomass and using forest waste.\u003c/p\u003e\u003cp\u003eThe use of plant-based biofuels significantly reduces the greenhouse effect: CO\u003csub\u003e2\u003c/sub\u003e emissions are close to zero, as the CO\u003csub\u003e2\u003c/sub\u003e released during combustion is used to produce organic matter during photosynthesis [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. The combustion of biofuels reduces emissions of sulfur compounds to the environment, and the ashes produced can be used to fertilize the soil [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. The development of biofuel or another environmentally friendly component production from plant biomass would contribute to the sustainability of agroecosystems, which is aimed at the circular economy [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eGrowing monocropped cultivations are still widespread in most countries to increase farm income, regardless of their negative environmental impacts [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Multicropped cultivations provide the opportunity to obtain a more varied multiple harvest during the vegetative season. It also has a positive effect on ecosystem services, helps to maintain and restore ecological balance through biodiversity, increases productivity and income, requires fewer inputs, reduces emissions, and increases the crop\u0026rsquo;s resilience to climatic shocks by increasing the C content of the soil [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Growing multicrops helps prevent the spread of weeds, diseases, and pests [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Growing crops together stimulates the activity of soil microorganisms, which accelerates humification processes [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The roots of multicrops are in different soil layers, allowing them to take up nutrients and water from different locations and thus supply them more efficiently. Growing together, plants of different species position their leaves at distinct height layers and spatial arrangements, enabling more efficient light capture, reduced mutual shading, and enhanced photosynthetic performance. [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Multicropping is also a promising strategy to increase land-use efficiency and cost-effectiveness of farming, especially for resource-constrained countries [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eTechnical hemp (\u003cem\u003eCannabis sativa\u003c/em\u003e L.) is a promising energy crop, one of the fastest-growing plants, and accumulates the most fiber in its stems [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Hemp typically produces between 5.5 and 8.5 t ha\u003csup\u003e-1\u003c/sup\u003e of dried biomass, depending on site climate, cultivar characteristics, and soil fertility [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Chaowana et al. [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e] found that hemp stems had desirable fuel properties: high volatile matter, high calorific value, low ash content, very low nitrogen content in the emissions, and undetectable sulfur. Technical hemp can be used in biorefineries to produce bioethanol, biodiesel, biohydrogen, organic acids, and other biomaterials [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Maize (\u003cem\u003eZea mays\u003c/em\u003e L.) is a versatile and high-yielding crop. Maize grains can be used to produce ethyl alcohol, stems and leaves can be used to produce paper, viscose, biochar, etc. Maize cultivation can produce on average 40\u0026ndash;60 t ha\u003csup\u003e-1\u003c/sup\u003e of green matter. Faba bean (\u003cem\u003eVicia faba\u003c/em\u003e L.) is a sustainable crop that not only provides ecological services but also leaves a large amount of residual biomass after the grain harvest, which can be used for energy production [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. A mixture of the above plants could also be a multifunctional crop, as technical hemp warrants high biomass yields and protection from pests and some diseases with low competition for sunlight. Maize ensures high crop productivity, and it has few diseases and pests in the Nordic countries. Faba beans provide ecological services to the crop by naturally providing companions with not only nitrogen but also potassium. In addition, the dried biomass residue capacity reaches 4\u003cb\u003e\u0026ndash;\u003c/b\u003e5 t ha\u003csup\u003e-1\u003c/sup\u003e in Lithuania.\u003c/p\u003e\u003cp\u003eWhat is the novelty and actuality of our research? First, ternary hemp, maize, and faba bean multifunctional crops for effective growth of abundant biomass have not yet been studied. The characteristics of their development (primarily as continuing cultivation for 3 years) in mixtures were not known. Second, the pesticide-free approach was used. Third, crops were grown for a short time, 103\u0026ndash;105 days after germination, until the faba beans were fully mature. This ensures an early supply of plant biomass for fuel production without loss of overall productivity. Other non-bean crops can grow for longer and provide a later biomass harvest for processing. Fourth, the energy and environmental balance of multicropping technologies was assessed, and a Life Cycle Assessment (LCA) of the produced fuel pellets was performed. Fifth, the technological parameters for producing fuel pellets from a ternary crop were registered in the Patent Office of the Republic of Lithuania as an invention.\u003c/p\u003e\u003cp\u003eThe aim of the present study was to assess the development indicators and productivity of separate crop species in the tested mixtures. Additionally, this study aims to highlight the relationships between crop development and productivity indices through innovative statistical analysis tools, and to summarize the energy use and environmental aspects of the researched technologies and produced pellets.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e"},{"header":"2. Materials and methods","content":"\u003ch2\u003e\u003cem\u003e2.1 Site description\u003c/em\u003e\u003c/h2\u003e\n\u003cp\u003eIn 2020\u0026ndash;2022, single, binary, and ternary crops of maize (\u003cem\u003eZea mays\u0026nbsp;\u003c/em\u003eL.), technical hemp (\u003cem\u003eCannabis sativa\u0026nbsp;\u003c/em\u003eL.), and faba bean (\u003cem\u003eVicia faba\u003c/em\u003e L.) were sown at the Experimental Station of Vytautas Magnus University Agriculture Academy (VMU AA) at the following coordinates: 54\u0026deg;53\u0026prime;7.5\u0026Prime; N 23\u0026deg;50\u0026prime;18.11\u0026Prime; E. The Experimental Station is located on the south-western side of Kaunas city, on the left bank of the Nemunas River, in the central part of Lithuania (Supplementary Fig. 1).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe soil at the experimental site is a silty loam (46% sand, 42% silt, 12% clay) Endohypogleyic-Eutric Planosol (Ple-gln-w) [21], with a texture of silty light loam on heavy loam. A soil pH\u003csub\u003eKC\u0026nbsp;\u003c/sub\u003ewas 7.3\u0026ndash;7.8, total nitrogen content \u0026ndash; 0.08\u0026ndash;0.13%, organic matter content \u0026ndash; 2.6\u0026ndash;2.9%, available potassium \u0026ndash; 118 mg\u0026middot;kg\u003csup\u003e-1\u003c/sup\u003e, available phosphorus \u0026ndash; 189\u0026ndash;280 mg\u0026middot;kg\u003csup\u003e-1\u003c/sup\u003e, available sulfur \u0026ndash; 1.2\u0026ndash;2.6 mg\u0026middot;kg\u003csup\u003e-1\u003c/sup\u003e, available magnesium \u0026ndash; 436\u0026ndash;790 mg\u0026middot;kg\u003csup\u003e-1\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eLithuania has sufficient precipitation in all seasons, especially in the warmer months, but recently, droughts have become more frequent in the summer. The average annual precipitation in Lithuania is 600\u0026ndash;900 mm, and evaporation is around 500 mm. The annual temperature is around 6.5\u0026ndash;7.5\u0026deg; C. During the experiment, meteorological conditions varied considerably each year. The air temperature in 2020 was close to the long-term average, with only July being slightly warmer. In that year, May and June had higher precipitation, but in April and July, which are particularly important for plant development and productivity, the crops experienced a lack of moisture (Supplementary Fig. 2). 2021 was a critical year for crop growth, with above-average temperatures and very low precipitation in June and July. In contrast, August was cooler compared to the other years of study and the long-term average, but with sufficient precipitation. In 2022, air temperature was lower than usual throughout the vegetation period, except at the end of the vegetation period, when the temperature in August was higher than the long-term average and there was a severe lack of humidity.\u003c/p\u003e\n\u003ch2\u003e\u003cem\u003e2.2. Experimental design and agronomic operations\u003c/em\u003e\u003c/h2\u003e\n\u003cp\u003eBefore setting up the experiment, oats (\u003cem\u003eAvena sativa\u003c/em\u003e L.) were grown in the experimental site. The experiment consisted of 7 treatments, 21 experimental plots in total, arranged with three replications in a randomized complete block design (RCBD). The size of the plot was 8 m\u003csup\u003e2\u003c/sup\u003e. Table 1 shows the list of performed treatments and their abbreviations.\u003c/p\u003e\n\u003cp\u003eMaize (\u003cem\u003eZea mays\u003c/em\u003e L.) (cultivar \u0026ldquo;Pioneer\u0026rdquo; hybrid P7034), technical hemp (\u003cem\u003eCannabis sativa\u003c/em\u003e L.) (cultivar \u0026ldquo;Austa SK\u0026rdquo;), and faba bean (\u003cem\u003eVicia faba\u003c/em\u003e L.) (cultivar \u0026ldquo;Vertigo\u0026rdquo;) as single, binary, and ternary crops, respectively, were grown to obtain the highest possible biomass yield at low-input pesticide-free conditions and by continuing the growth of crops.\u003c/p\u003e\n\u003cp\u003eThe soil of the experiment was plowed with a Gamega PP-3-43 plow (Lithuanian producer) with semi-helical shellboards in each fall (usually in October) and cultivated with a complex Laumetris KLG-3.6 cultivator (Lithuanian producer) in spring (usually in April) before sowing. Before sowing, mineral complex fertilizer NPK 15:15:15 (300 kg\u0026middot;ha\u003csup\u003e\u0026minus;1\u003c/sup\u003e) was treated by hand equipment. This fertilization rate is insufficient for achieving high biomass yields; however, we aim to demonstrate a more pronounced effect of continued multicropping on soil fertility. We expected that the ecological services of faba beans would compensate for the negative impact of crops continuing to grow on the soil properties. Experimental plots were sown by hand equipment according to specially created seed distributing schemes [22] (Supplementary Fig. 3).\u0026nbsp;The presented plant sowing schemes ensured the development of crop mixtures with minimal competition (Supplementary Fig. 4).\u003c/p\u003e\n\u003cp\u003eSowing rates depend on the scheme and are presented in more detail in Supplementary Table 1. The crops\u0026rsquo; inter-rows were loosened 1\u0026ndash;2 times until the crops covered the inter-rows. Crop biomass production was harvested after a short 103\u0026ndash;105-day vegetative season, when the faba beans reached full maturity (Supplementary Table 2). At that time, the biomass of maize and hemp had not yet reached its maximum. Still, faba beans produce abundant and valuable biomass (especially in the first year of growing), which we did not want to lose because in August, there is usually a shortage of biomass for fuel production in Lithuania. The applied technological operations and technical means, and their energy and CO\u003csub\u003e2eq.\u0026nbsp;\u003c/sub\u003ebalance are discussed in more detail in our previous article [23].\u003c/p\u003e\n\u003ch2\u003e \u003cem\u003e2.3. Methods and analysis\u003c/em\u003e\u003c/h2\u003e\n\u003cp\u003eTo determine crop development indicators, measurements were made during the vegetation period when faba bean began to bloom (maize BBCH 51\u0026ndash;53, technical hemp BBCH 60\u0026ndash;62, faba bean BBCH 63\u0026ndash;65), assessing plant height, leaf chlorophyll index, assimilation area, and green (fresh) biomass of each crop species. At the end of the plant vegetation period (maize BBCH 79\u0026ndash;80, technical hemp BBCH 81\u0026ndash;83, faba bean BBCH 86\u0026ndash;88), not only was the green biomass of maize, hemp, and faba bean assessed separately, but also the total green and dried biomass of the crops per unit area.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFor development indicator studies, five plants of each crop species were cut out in each experimental plot. The assimilation area of plant leaves (cm\u003csup\u003e2\u003c/sup\u003e) was determined with a leaf area meter Win Dias (\u0026ldquo;Delta-T Devices\u0026rdquo; Ltd, UK). The chlorophyll index of plant leaves was measured with a chlorophyll meter CCM\u0026ndash;200 Plus. The height of the plants was measured, and they were weighed, thus determining their green biomass. At the end of the vegetation period, samples were taken at least five spots per plot, in a 0.5 m longitudinal row. An average sample was formed. A total of 36 research samples were formed. To determine the dried biomass, the samples were dried in a thermostat at a temperature of 105 \u0026deg;C.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe plant biomass grown can be used in energy and other industries, especially in the production of biofuels, fertilizers, bio-additives, etc.; therefore, its elemental composition is important (Supplementary Table 3). In Supplementary Table 4, the main biofuel pellet characteristics are presented. The chemical composition was tested in the laboratories of the Lithuanian Research Centre of Agriculture and Forestry in Kaunas, Lithuania.\u003c/p\u003e\n\u003cp\u003eThe experimental data were analyzed using one-way ANOVA, and the treatment effect was estimated by the F-test and the least significant difference (LSD). SYSTAT and SELEKCIJA software were employed. Significant differences between the treatments are marked with different lowercase letters, when\u0026nbsp;\u003cem\u003eP\u0026le;0.05\u0026gt;0.01\u003c/em\u003e. Significant differences between the vegetation conditions of individual years of the experimentation are marked with different uppercase letters, when\u0026nbsp;\u003cem\u003eP\u0026le;0.05\u0026gt;0.01\u003c/em\u003e. A correlation analysis was performed with SigmaStat software.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ePrincipal Component Analysis (PCA) was used to establish the correlation between different indicators in single, binary, and ternary crops. This analysis creates new artificial variables (principal components) based on the analysed variables. Its central premise was the ability to visualise the relationships between individual variables in a two-dimensional diagram showing the coordinate system of the first two principal components. Based on the position of the vectors in space, it is possible to determine which variables are correlated with each other. A lower angle between the vectors shows the stronger positive correlation. When the vectors are aligned on the same line but in opposite directions, there is a strong negative correlation between the variables. However, when the vectors are aligned at an angle close to 90 degrees, there is no correlation [24].\u003c/p\u003e\n\u003cp\u003eThe results of the investigations were also grouped using cluster analysis. The clustering of all tested single, binary, and ternary crops, considering the estimated average height of crops, leaf assimilation area, and chlorophyll index, crop green (fresh) and dried biomass, was carried out according to the Ward criterion using Euclidean distance matrices. Statistical analysis was performed using the Statistica software package Statistica 10 (TIBCO Software Inc., Palo Alto, CA, USA). Calculations were performed at Vytautas Magnus University Agriculture Academy.\u003c/p\u003e"},{"header":"3. Results and discussion","content":"\u003ch2\u003e\u003cem\u003e3.1. Average plant height\u003c/em\u003e\u003c/h2\u003e\n\u003cp\u003eThe intensity of crop diversification (factor A) negatively affected the average heights of maize and technical hemp. They were significantly higher when grown as single crops, and the lowest when grown as a ternary crop. Maize and technical hemp were also taller when grown together with faba bean (Table 2). Fisher et al. [25] in Germany and Hirpa [26] in Ethiopia found no effect on maize height when grown in a binary crop with faba bean. Li et al. [27] found the negative effect. The height indicators of faba bean differed from those of maize and technical hemp. The lowest faba bean height was observed in a single crop, with a significant difference of 10.9 cm when grown together with maize.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eRegarding the effect of crop continuing growing (factor B) on plant height, it was found that the lowest height of maize and technical hemp was in the last year of the experiment, i.e., the heights were 1.6\u0026ndash;1.9 times less because of degradation of most soil properties [28].\u003c/p\u003e\n\u003cp\u003eA comprehensive assessment of the different indicators studied showed that plant height in a hemp crop was the highest expressed at crop development and productivity level compared to other experimental treatments [29] (Supplementary Fig. 5).\u003c/p\u003e\n\u003ch2\u003e\u003cem\u003e3.2. \u0026nbsp;Leaf assimilation area\u003c/em\u003e\u003c/h2\u003e\n\u003cp\u003eThe increase in the number of plant species in the crop influenced the decrease in the assimilation area of their leaves. The largest leaf area of maize and technical hemp was determined when they grew as single crops (Table 3).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eWhen maize grew together with faba bean, their leaf assimilation area was 16% higher than in other diversified crops. This is likely because faba beans offer less competition for maize growth and supplement the soil with nitrogen, a nutrient essential for plant growth. Similarly, Li et al. [30] and Yang et al. [31] also found that the maize-soybean binary crop increased the assimilation area of maize leaves. Yang et al. [32] and Corre-Hellou et al. [33] confirmed these results with pea (\u003cem\u003ePisum sativum\u003c/em\u003e L.) cover- and inter-crops. A comprehensive data analysis revealed that maize leaves\u0026rsquo; assimilation area was the highest at crop development and productivity levels compared to other experimental treatments (Supplementary Fig. 5). Additionally, we found a positive moderate relationship between maize leaf assimilation area and plant height (r = 0.64, \u003cem\u003eP\u003c/em\u003e \u0026le; 0.050 \u0026gt; 0.010).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe largest leaf assimilation area of hemp was found in the binary crop with faba bean, and the highest area of faba bean leaves was found in the binary crop with maize. On the contrary, Wu et al. [34] found that when faba bean grows together with maize, increased competition for sunlight causes faba bean stems to lengthen and reduces their leaf assimilation area.\u003c/p\u003e\n\u003cp\u003eA significant effect of vegetative conditions on the assimilation area of maize and technical hemp leaves was found. In the second year of the study, the assimilation area of maize and technical hemp leaves was significantly higher. Contrary to expectations, the hot and dry months of June-July in that year did not harm the development of these crops. Different vegetation conditions in individual years of the study influenced a consistent increase in the assimilation area of faba bean leaves.\u003c/p\u003e\n\u003ch3\u003e\u003cem\u003e3.3.\u0026nbsp;\u003c/em\u003e\u003cem\u003eChlorophyll index of plant leaves\u003c/em\u003e\u003c/h3\u003e\n\u003cp\u003eThe chlorophyll index of leaves of each single crop was usually higher than that of diversified crops (Table 4). Similarly, Pe\u0026ntilde;afiel\u0026ndash;Sandova [35] also found the highest chlorophyll index in the maize single crop.\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eAccording to the average data of our experiment, factor B, the chlorophyll index in the last year of the experiment decreased 3 times in maize and 1.4 times in technical hemp crops compared to the first year of the study. The opposite results were obtained in faba bean crops, since the chlorophyll index was significantly highest in the third year of the study.\u003c/p\u003e\n\u003cp\u003eAccording to the comprehensive analysis of the research data, the crop leaf chlorophyll index was the most common in the tested ternary crop. In another cultivation, the rating of this index mainly did not exceed the evaluation threshold (Supplementary Fig. 5).\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003e\u003cem\u003e3.4. Green biomass of individual plant species\u003c/em\u003e\u003c/h2\u003e\n\u003cp\u003eAn interaction was found between the effect of crop diversification and vegetation conditions of individual research years on the average green biomass of plants at the end of the vegetation. Crop diversification negatively affected the green biomass of individual plants, since it was highest in single crops of maize, technical hemp, and faba bean (Tables 5 and 6).\u003c/p\u003e\n\u003cp\u003eAmong the diversified crops, the highest green biomass of maize and technical hemp was found when they grew together with faba bean. Here, the green biomass of maize was 3 times higher, and that of technical hemp was 2 times\u0026nbsp;higher than in the ternary crop. Combining maize with various legumes, such as faba bean, improved maize crop growth, increased its green biomass, and increased plant quality [36]. The green biomass of faba bean did not differ significantly\u0026nbsp;in the diversified crops. The vegetation conditions of the research years also had a significant impact on the green biomass of maize, technical hemp, and faba bean. The highest green biomass of these plants was in the first year of the experiment, and then it consistently decreased. In the third year of the experiment, the green biomass of maize, hemp, and faba bean decreased by 2\u0026ndash;4 times.\u003c/p\u003e\n\u003cp\u003eMaize green biomass was partly dependent on its leaf chlorophyll index (r = 0.58, \u003cem\u003eP\u0026nbsp;\u003c/em\u003e\u0026le; 0.050 \u0026gt; 0.010) and leaf assimilation area (r = 0.72, \u003cem\u003eP \u0026le;\u0026nbsp;\u003c/em\u003e0.050 \u0026gt; 0.010). Technical hemp dependencies were similar (r = 0.87, r = 0.86, \u003cem\u003eP\u003c/em\u003e \u0026le; 0.010 \u0026gt; 0.001). A statistically significant correlation was also established between faba bean leaf assimilation area and capacity of green biomass (r = 0.87, \u003cem\u003eP\u003c/em\u003e \u0026le; 0.010 \u0026gt; 0.001). Plants\u0026rsquo; green biomass also partly depended on the crop density and average height of plants.\u003c/p\u003e\n\u003ch2\u003e\u003cem\u003e3.5. Total biomass of cultivations\u003c/em\u003e\u003c/h2\u003e\n\u003cp\u003eAn interaction was established between the effect of crop diversification and the vegetative conditions of individual research years on the average green biomass of crops at the end of the plant vegetation. Among single crops, the highest green biomass per unit area was observed for maize (4,410.7 g m\u003csup\u003e-2\u003c/sup\u003e). Among diversified crops, the highest green biomass was obtained when growing a binary crop of maize and faba bean (3,421.5 g m\u003csup\u003e-2\u003c/sup\u003e). Shtaya et al. [37] also confirmed that faba bean, when mixed with other crops, increased the total green biomass of the crop. Streit et al. [38] obtained the opposite results and claim that faba bean in mixtures produces an average of 5% more green biomass than in single crops. In our experiment, the green biomass of the ternary crop was 25% lower than that of the binary crop of maize and faba bean (Table 7).\u003c/p\u003e\n\u003cp\u003eThe vegetative conditions of individual research years also had a significant effect on the green biomass of crops at the end of the plant vegetation period. It was significantly highest in the first year of the experiment and decreased by 30% in the second and third research years.\u003c/p\u003e\n\u003cp\u003eAs expected, the ternary crop had the highest total dried biomass, which was 8 times higher than that of the single maize crop. This is because at the time of biomass harvesting, maize contained a large amount of water, while the faba bean biomass was close to air-dried (BBCH 95\u0026ndash;97). According to Ciampitti et al. [39], in faba bean, the highest N\u003csub\u003e2\u003c/sub\u003e fixation occurs at the flowering stage. Therefore, harvesting at the right time is very important not only to ensure the best crop yield and the highest possible N\u003csub\u003e2\u003c/sub\u003e contribution to subsequent crops, but also to maximize biomass energy yield. This was observed in our experiment, where maize benefited from the ecological services provided by faba bean. Faba beans, together with companions, can also increase not only total yield and income, but reduce crop weediness and disease, increase land use efficiency, and thus increase crop sustainability [37, 40]. Moreover, the dried biomass of all single crops was lower compared to the biomass of diversified crops (Table 8). Dzvene [41] obtained similar results.\u003c/p\u003e\n\u003cp\u003eThe crops\u0026rsquo; continued growth harmed their total biomass yields. Only in the first year of the study did we receive the highest yields (especially in the ternary crop), and in the second and last years of the study, they decreased by about 6.5 times due to the degradation of soil properties. Bybee\u0026ndash;Finley et al. [42] found that in an experiment with millet, sorghum, and hemp, in the first year of crop cultivation, the biomass of plants grown in mixtures was higher than that of single crops.\u003c/p\u003e\n\u003cp\u003eAfter conducting a comprehensive assessment of the experimental data, it was found that the ternary crop had the highest CEI value; therefore, it was the most effective in growing plant total biomass. The formation of higher dried biomass in the ternary crop was most influenced by the higher leaf chlorophyll index (Supplementary Fig. 5, marked in yellow). A higher CEI value was also obtained in the binary M+H cultivation. This confirms the statement of Branca et al. [43] and Hu et al. [44] that maize and technical hemp are the most promising crops in the field of biomass processing. A generalized cluster analysis of our experimental data showed that maize and hemp were more efficiently grown in intercropping with faba beans due to their higher biomass capacity (Fig. 1).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe PCA was also done for different species of companion crops in cultivation:\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eMaize.\u003c/em\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eIn 2020, the aboveground biomass of maize in the middle and end of the vegetative season was closely related to plant height and plant photosynthetic indicators ˗ plant leaf assimilation area and chlorophyll index in leaves (Fig. 2). In 2021, the aboveground biomass of maize in the middle of the vegetation correlated with plant leaf assimilation area and chlorophyll index in leaves. The latter indicators had less influence on the aboveground biomass of plants at the end of the vegetation. In 2022, similar trends were found as in 2021.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eTechnical hemp.\u003c/em\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eIn 2020, the aboveground biomass of hemp in the middle and end of the vegetative period was most influenced by plant average height. The influence of leaf assimilation area was lower (Fig. 3). In 2021, two groups of closely correlated indicators emerged: the first group consisted of the aboveground biomass of plants at the beginning of the vegetative period and the leaf assimilation area, and the second group consisted of the aboveground biomass of plants at the end of the vegetative period and plant height. In 2022, the aboveground biomass of hemp in the middle of the vegetative season was closely related to the plant leaf assimilation area. The aboveground biomass at the end of the vegetative period was more dependent on the plant height.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eFaba bean.\u003c/em\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eIn 2020, the aboveground biomass of faba beans in the middle and end of the vegetative period was closely related to plant photosynthetic indicators \u0026ndash; plant leaf assimilation area and chlorophyll index (Fig. 4). In 2021, the aboveground biomass of faba beans at the middle of the vegetative period was less influenced by plant height and photosynthetic indicators. In 2022, the aboveground biomass of faba beans at the end of the vegetative period depended most on the chlorophyll concentration in the leaves.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;In summary, it can be stated that crop diversification had a positive impact on the total yield of dried biomass. The dried biomass of the ternary crop was 4\u0026ndash;8 times higher compared to single crops, and 2 times higher than that of binary crops. It is recommended to grow low-fertilized and pesticide-free ternary crops in existing on-farm crop rotations for one year only. Continuing crops need to be well fertilized, because biomass productivity gradually decreases. Aubin et al. [45] concluded that higher N rates can maximally improve hemp growth, plant height, and biomass.\u003c/p\u003e\n\u003ch2\u003e\u003cem\u003e3.6. Technological, energy, and environmental aspects\u003c/em\u003e\u003c/h2\u003e\n\u003cp\u003eAll tested cropping technologies include stubble cultivation (depth 12\u0026ndash;15 cm) in fall, deep plowing before wintering, pre-sowing cultivation, and fertilization. One-pass conventional sowing was used for single crops, while two-pass ternary sowing was used for binary and ternary crops. Inter-row loosening (2\u0026ndash;3 cm depth) was performed twice for all cultivations, except the ternary crop. In the ternary crop, the inter-row loosening was performed once due to higher crop densities. In calculations, we simulated one-pass biomass harvesting (low harvester load) for M, FB, and M+FB crops, one-pass biomass harvesting (high harvester load) for H, M+H, and H+FB crops, and two-pass biomass harvesting (high harvester load) for the ternary crop. Tractor power (kW) for calculations ranged from 45\u0026ndash;67 kW (for sowing, fertilization, and shallow loosening) to 102 kW (for deeper tillage operations). The combine harvester power remained constant at 250 kW. We adjusted the field capacity from 1.82 to 0.68 ha/harvester load. Technological aspects are discussed in more detail in Romaneckas et al. [23].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe highest consumption of fuel was calculated for the ternary crop (M+H+FB) (103.3 L ha\u003csup\u003e\u0026minus;1\u003c/sup\u003e) due to a higher number and more powerful operations (Supplementary Table 5). Maize and faba bean single cropping used the lowest amount of fuel. Although the technology for growing the ternary crop required more energy input, the yield obtained compensated for this. So, the highest net energy (367,668.1 MJ ha\u003csup\u003e\u0026minus;1\u003c/sup\u003e) was also obtained in the ternary crop. Obviously, growing more plant companions in the same crop results in higher net energy [46].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe lowest total CO\u003csub\u003e2eq.\u003c/sub\u003e (greenhouse gases) emissions were calculated for single hemp and faba bean cultivations, and the highest were for M+H and M+FB binary crops. \u0026nbsp;GHG emission of ternary crop was average (1,541.90 kg ha\u003csup\u003e\u0026minus;1\u003c/sup\u003e CO2\u003csub\u003eeq.\u003c/sub\u003e) [23].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe short-growing biomass of the tested maize-hemp-faba bean ternary crop can be effectively used for energy purposes and crop fertilization, because the pH is close to neutral, contains about 1% of\u0026nbsp;nitrogen and potassium, and about 0.2%\u0026nbsp;of phosphorus (Supplementary Table 2), and the average yield of total dried biomass reaches almost 20 t ha\u003csup\u003e-1\u003c/sup\u003e. A National invention patent had been registered to produce pellets from the biomass of the ternary crop discussed [47]. These pellets had a higher density than those made from biomass of other cultivations, one of the lowest ash contents, and the highest ash shrinkage starting temperature. They met the requirements of the ISO 17225-6:2021 standard (Supplementary Table 3). Moreover, the burning of pellets in household low-power boilers did not have negative environmental consequences [48]. Unlike the evaluation of crop production technologies, the Life Cycle Assessment (LCA) of the impact of pellet disposition and use (transportation, heat production, and ash utilization) on abiotic depletion, global warming potential, acidification, and eutrophication indices showed that binary M+H crops had the lowest impact on the environment. Pellets made from ternary crop biomass had an average dimension [49].\u0026nbsp;\u003c/p\u003e"},{"header":"4. Conclusions","content":"\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eDue to the eco-service of faba beans, maize and technical hemp mixtures with faba beans tended to have 14% greater height, 24% higher leaf assimilation area, and 19% higher chlorophyll index. With the increase in the number of plant species in the crop mixtures, the biomass productivity of each plant species decreased, but the total biomass productivity per plot area increased. In the first year of the experiment, a ternary maize\u0026ndash;technical hemp\u0026ndash;faba bean crop produced 4\u0026ndash;8 times more dried biomass than individual single species. Multicropping continuation for the next two years decreased biomass yields by up to 11 times. Ternary multicrop productivity reached the highest CEI value (4.54), which was mainly influenced by the chlorophyll index of the plant leaves. Ternary cultivation was the most fuel-consuming technology (18\u0026ndash;32% higher fuel consumption) \u0026ndash; 103.3 L ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e due to higher energy input. However, it could be characterized by the most significant net energy (367,668.1 MJ ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) because of the most abundant yield of dried biomass. The pellets produced from ternary crop biomass met the standards, had the highest density (1,238 kg m\u003csup\u003e-3\u003c/sup\u003e), one of the lowest ash contents (6%), and the highest ash shrinkage starting temperature (1042\u0026deg;C). The LCA of pellet disposition and use showed that the binary M\u0026thinsp;+\u0026thinsp;H crop had the lowest impact on the environment. However, biomass growing technology was one of the most contaminating; therefore, it is advisable to develop high-capacity yielding ternary crops, whose GHG emissions and LCA effects on the produced pellets had an average dimension.\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eContributions\u003c/strong\u003e\u003cp\u003eConceptualization, K.R., J.B.; methodology, K.R., J.B.; software, J.B.; validation, J.B., K.R.; formal analysis, J.B., K.R., A.M.; investigation, J.B., R.K., A.S. and K.R.; resources, J.B, R.K., A.S. and K.R.; data curation, J.B.; writing\u0026mdash;original draft preparation, J.B., K.R.; writing\u0026mdash;review and editing, K.R., J.B., R.K. and A.S; visualization, J.B., K.R., A.M.; supervision, K.R. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eContributions:\u003c/strong\u003e Conceptualization, K.R., J.B.; methodology, K.R., J.B.; software, J.B.; validation, J.B., K.R.; formal analysis, J.B., K.R., A.M.; investigation, J.B., R.K., A.S. and K.R.; resources, J.B, R.K., A.S. and K.R.; data curation, J.B.; writing\u0026mdash;original draft preparation, J.B., K.R.; writing\u0026mdash;review and editing, K.R., J.B., R.K. and A.S; visualization, J.B., K.R., A.M.; supervision, K.R. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest:\u003c/strong\u003e The authors declare that they have no known competing financial interests or personal relationships that may influence the work reported in this paper.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e The reported work in this article was partially supported by the Ministry of Education, Science and Sports of the Republic of Lithuania and Research Council of Lithuania (LMTLT) under the Program \u0026lsquo;University Excellence Initiative\u0026rsquo; Project \u0026lsquo;Development of the Bioeconomy Research Center of Excellence\u0026rsquo; (BioTEC), agreement No S-A-UEI-23-14.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials:\u003c/strong\u003e described data and materials are available from the corresponding author upon request. The methodology for the comprehensive assessment and a detailed calculation example is provided in our previous article Kimbirauskienė R, Sinkevičienė A, \u0026Scaron;vereikaitė A, Romaneckas K. The Complex Effect of Different Tillage Systems on the Faba Bean Agroecosystem. Plants. 2024; 13(4):513. https://doi.org/10.3390/plants13040513, Supplementary materials, Table S1. The meteorological indicators presented in the article were obtained from the Lithuanian Hydrometeorological Service, Kaunas Meteorological Station. https://www.meteo.lt/en/organization/structure-and-contacts/contacts/\u0026quot; in Supplementary materials, Figure 2. Data on main characteristics of grown multi crop solid fuel pellets was referred from Petlickaitė, R., Jasinskas, A., Domeika, R., Pedi\u0026scaron;ius, N., Lemanas, E., Praspaliauskas, M., Kukharets, S. Evaluation of the Processing of Multi-Crop Plants into Pelletized Biofuel and Its Use for Energy Conversion, Processes. 11(2), 421, https://doi.org/10.3390/pr11020421 (2023) in Supplementary materials, Table 3.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eOishy, M. N. et al. A. Unravelling the effects of climate change on the soil-plant-atmosphere interactions: A critical review. SEH. 10130, (2025). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.seh.2025.100130\u003c/span\u003e\u003cspan address=\"10.1016/j.seh.2025.100130\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eErdiwansyah, A. G. et al. Prospects for renewable energy sources from biomass waste in Indonesia. Case Stud. \u003cem\u003eChem. Environ. Eng.\u003c/em\u003e \u003cb\u003e10\u003c/b\u003e, 100880. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cscee.2024.100880\u003c/span\u003e\u003cspan address=\"10.1016/j.cscee.2024.100880\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRoy, R. et al. Steam explosion treated biomass as a renewable fuel source: A review from collection to combustion. \u003cem\u003eFuel\u003c/em\u003e \u003cb\u003e378\u003c/b\u003e, 132883. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.fuel.2024.132883\u003c/span\u003e\u003cspan address=\"10.1016/j.fuel.2024.132883\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLicata, M. et al. A comparative study to assess the production of two oilseed crops (Brassica carinata A. Braun and Carthamus tinctorius L.) and the energy potential of their agricultural biomass residues. \u003cem\u003eHeliyon\u003c/em\u003e \u003cb\u003e10\u003c/b\u003e (22), e38654. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.heliyon.2024.e38654\u003c/span\u003e\u003cspan address=\"10.1016/j.heliyon.2024.e38654\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJasinskas, A., Petlickaitė, R., Praspaliauskas, M., Romaneckas, K. \u0026amp; Sinkevičienė, A. Impact of ash obtained after multi-crop pellet burning on spring barley fertilization. In:Engineering for Rural Development: 23rd International Scientific Conference, ERDev 2024-Proceedings. Latvia University of Life Sciences and Technologies, 23, pp. 644\u0026ndash;649 (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMuthulakshmi, C., Sivaranjani, R. \u0026amp; Selvi, S. Modification of sesame (Sesamum indicum L.) for Triacylglycerol accumulation in plant biomass for biofuel applications. \u003cem\u003eBiotechnol. Rep.\u003c/em\u003e \u003cb\u003e32\u003c/b\u003e, e00668. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.btre.2021.e00668\u003c/span\u003e\u003cspan address=\"10.1016/j.btre.2021.e00668\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWang, T. et al. From intercropping to monocropping: The effects of Pseudomonas strain to facilitate nutrient efficiency in peanut and soil. \u003cem\u003ePlant. Physiol. Biochem.\u003c/em\u003e \u003cb\u003e219\u003c/b\u003e, 109378. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.plaphy.2024.109378\u003c/span\u003e\u003cspan address=\"10.1016/j.plaphy.2024.109378\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKumar, R. K. S., Sasikumar, R. \u0026amp; Dhilipkumar, T. Exploiting agro-waste for cleaner production: A review focusing on biofuel generation, bio-composite production, and environmental considerations. \u003cem\u003eJ. Clean. Prod.\u003c/em\u003e \u003cb\u003e435\u003c/b\u003e, 140536. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jclepro.2023.140536\u003c/span\u003e\u003cspan address=\"10.1016/j.jclepro.2023.140536\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAlcon, F. et al. Cost benefit analysis of diversified farming systems across Europe: Incorporating non-market benefits of ecosystem services. \u003cem\u003eSci. Total Environ.\u003c/em\u003e \u003cb\u003e912\u003c/b\u003e, 169272. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.scitotenv.2023.169272\u003c/span\u003e\u003cspan address=\"10.1016/j.scitotenv.2023.169272\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHashakimana, L., Tessema, T., Niyitanga, F., Cyamweshi, A. R. \u0026amp; Mukuralinda, A. Comparative analysis of monocropping and mixed cropping systems on selected soil properties, soil organic carbon stocks, and simulated maize yields in drought-hotspot regions of Rwanda. \u003cem\u003eHeliyon\u003c/em\u003e \u003cb\u003e9\u003c/b\u003e, e19041. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.heliyon.2023.e19041\u003c/span\u003e\u003cspan address=\"10.1016/j.heliyon.2023.e19041\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eL\u0026eacute;onidas, H. et al. Monocropping vs mixed cropping systems under a changing climate: Smallholder farmers' perceptions and farm profitability in Eastern Rwanda. \u003cem\u003eEnviron Sustain Ind\u003c/em\u003e. \u003cb\u003e24\u003c/b\u003e, 100527. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.indic.2024.100527\u003c/span\u003e\u003cspan address=\"10.1016/j.indic.2024.100527\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eSu, B., Liu, X., Cui, L., Xiang, B. \u0026amp; Yang, W. Suppression of Weeds and Increases in Food Production in Higher Crop Diversity Planting Arrangements: A Case Study of Relay Intercropping. \u003cem\u003eCrop Sci.\u003c/em\u003e \u003cb\u003e58\u003c/b\u003e (4), 1729\u0026ndash;1739. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2135/cropsci2017.11.0670\u003c/span\u003e\u003cspan address=\"10.2135/cropsci2017.11.0670\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eTariq, A. et al. Combining different species in restoration is not always the right decision: Monocultures can provide higher ecological functions than intercropping in a desert ecosystem. \u003cem\u003eJ. Environ. Manag\u003c/em\u003e. \u003cb\u003e357\u003c/b\u003e, 120807. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jenvman.2024.120807\u003c/span\u003e\u003cspan address=\"10.1016/j.jenvman.2024.120807\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChimonyo, V. G. P. et al. Chapter 18 - Yield and water use gaps in cereal multicrop systems in sub-Saharan Africa under climate change. Food Processing Technology. Principles and Practice. A volume in Woodhead Publishing Series in Food Science, Technology and Nutrition, pp. 313\u0026ndash;329 (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eVanino, S. et al. Rarina, R. A comprehensive assessment of diversified cropping systems on agro-environmental sustainability in three Mediterranean long-term field experiments. \u003cem\u003eEur. J. Agron.\u003c/em\u003e \u003cb\u003e140\u003c/b\u003e, 126598. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.eja.2022.126598\u003c/span\u003e\u003cspan address=\"10.1016/j.eja.2022.126598\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eMamimin, C., O-Thong, S. \u0026amp; Reungsang, A. Enhancing biogas production from hemp biomass residue through hydrothermal pretreatment and co-digestion with cow manure: Insights into methane yield, microbial communities, and metabolic pathways. \u003cem\u003eJ. Environ. Manag\u003c/em\u003e. \u003cb\u003e370\u003c/b\u003e, 123039. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jenvman.2024.123039\u003c/span\u003e\u003cspan address=\"10.1016/j.jenvman.2024.123039\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eV\u0026aacute;vrov\u0026aacute;, K. et al. Economic evaluation of Hemp\u0026rsquo;s (Cannabis sativa) residual biomass for production of direct energy or biochar. \u003cem\u003eFuel\u003c/em\u003e \u003cb\u003e329\u003c/b\u003e, 125435. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.fuel.2022.125435\u003c/span\u003e\u003cspan address=\"10.1016/j.fuel.2022.125435\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eChaowana, P. et al. Utilization of hemp stalk as a potential resource for bioenergy. \u003cem\u003eMat. Sci. En Tech.\u003c/em\u003e \u003cb\u003e7\u003c/b\u003e, 19\u0026ndash;28. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.mset.2023.07.001\u003c/span\u003e\u003cspan address=\"10.1016/j.mset.2023.07.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eTripathi, M. et al. Conversion technologies for valorization of hemp lignocellulosic biomass for potential biorefinery applications. \u003cb\u003e320\u003c/b\u003e, 124018, (2023). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.seppur.2023.124018\u003c/span\u003e\u003cspan address=\"10.1016/j.seppur.2023.124018\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eG\u0026oacute;mez, L. D. et al. Caruso, L. Valorising faba bean residual biomass: Effect of farming system and planting time on the potential for biofuel production. \u003cem\u003eBiomass Bioenerg\u003c/em\u003e. \u003cb\u003e107\u003c/b\u003e, 227\u0026ndash;232. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.biombioe.2017.10.019\u003c/span\u003e\u003cspan address=\"10.1016/j.biombioe.2017.10.019\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eIUSS Working Group WRB. World Reference Base for Soil Resources: International soil classification system for naming soils and creating legends for soil maps, 4th ed., Vienna, Austria, (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRomaneckas, K. et al. Short-Term Impact of Multi-Cropping on Some Soil Physical Properties and Respiration. \u003cem\u003eAgron\u003c/em\u003e. 12, 141, (2022). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agronomy12010141\u003c/span\u003e\u003cspan address=\"10.3390/agronomy12010141\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eRomaneckas, K., Švereikaitė, A., Kimbirauskienė, R., Sinkevičienė, A. \u0026amp; Balandaitė, J. The Energy and Environmental Evaluation of Maize, Hemp and Faba Bean Multi-Crops. \u003cem\u003eAgron\u003c/em\u003e \u003cb\u003e13\u003c/b\u003e, 2316. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agronomy13092316\u003c/span\u003e\u003cspan address=\"10.3390/agronomy13092316\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJolliffe, I. T. \u0026amp; Cadima, J. Principal component analysis: a review and recent developments. Philosophical Transactions of the Royal Society. \u003cem\u003eMath. Phys. Eng. Sci.\u003c/em\u003e \u003cb\u003e374\u003c/b\u003e, 1\u0026ndash;16. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1098/rsta.2015.0202\u003c/span\u003e\u003cspan address=\"10.1098/rsta.2015.0202\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eFisher, J., Bohm, H. \u0026amp; Heβ, J. Maize\u0026ndash;bean intercropping yields in Northern Germany are comparable to those of pure silage maize. \u003cem\u003eEur. J. Agron.\u003c/em\u003e \u003cb\u003e112\u003c/b\u003e, 125\u0026ndash;147. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.eja.2019.125947\u003c/span\u003e\u003cspan address=\"10.1016/j.eja.2019.125947\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHirpa, T. Effect of intercrop row arrangement on maize and haricot bean productivity and the residual soil. \u003cem\u003eGJSFR\u003c/em\u003e \u003cb\u003e14\u003c/b\u003e, 1\u0026ndash;9 (2014).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLi, Z. et al. The synergistic priming effect of exogenous salicylic acid and H2O2 on chilling tolerance enhancement during maize (\u003cem\u003eZea mays\u003c/em\u003e L.) seed germination. \u003cem\u003eFront. Plant. Sci.\u003c/em\u003e \u003cb\u003e8\u003c/b\u003e, 1153. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fpls.2017.01153\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2017.01153\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBalandaitė, J. Crop biodiversity influence on multi-crops sustainability, productivity and energy efficiency. Doctoral Thesis. Vytautas Magnus University, Kaunas, Lithuania, (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBalandaitė, J., Romaneckas, K., Kimbirauskienė, R. \u0026amp; Sinkevičienė, A. Comprehensive Assessment of the Effect of Multi-Cropping on Agroecosystems. \u003cem\u003ePlants\u003c/em\u003e \u003cb\u003e13\u003c/b\u003e (10), 1372. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/plants13101372\u003c/span\u003e\u003cspan address=\"10.3390/plants13101372\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eLi, Y. et al. Maize\u0026ndash;soybean relay cropping increases soybean yield synergistically by extending the post-anthesis leaf stay-green period and accelerating grain filling. \u003cem\u003eCrop J.\u003c/em\u003e \u003cb\u003e11\u003c/b\u003e (6), 1921\u0026ndash;1930. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cj.2023.05.011\u003c/span\u003e\u003cspan address=\"10.1016/j.cj.2023.05.011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYang, Z. et al. GmPTF1 modifies root architecture responses to phosphate starvation primarily through regulating GmEXPB2 expression in soybean. \u003cem\u003ePlant. J.\u003c/em\u003e \u003cb\u003e107\u003c/b\u003e, 525\u0026ndash;543. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/tpj.15307\u003c/span\u003e\u003cspan address=\"10.1111/tpj.15307\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eYang, H. et al. Yield photosynthesis and leaf anatomy of maize in inter- and mono-cropping systems at varying plant densities. \u003cem\u003eCrop J.\u003c/em\u003e \u003cb\u003e10\u003c/b\u003e, 893\u0026ndash;903 (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCorre-Hellou, G., Fustec, J. \u0026amp; Crozat, Y. Interspecific competition for soil N and its interaction with N2 fixation, leaf expansion and crop growth in pea\u0026ndash;barley intercrops. \u003cem\u003ePlant. Soil.\u003c/em\u003e \u003cb\u003e282\u003c/b\u003e, 95\u0026ndash;208 (2006).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eWu, Y. et al. Combining modelling and experiment to quantify light interception and inter row variability on intercropped soybean in strip intercropping. \u003cem\u003eEur. J. Agron.\u003c/em\u003e \u003cb\u003e164\u003c/b\u003e, 127508. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.eja.2025.127508\u003c/span\u003e\u003cspan address=\"10.1016/j.eja.2025.127508\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2025).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePe\u0026ntilde;afiel\u0026ndash;Sandova, Z. B. Effect of urea on lead absorption in corn (\u003cem\u003eZea mays\u003c/em\u003e L), spinach (\u003cem\u003eSpinacia oler\u0026aacute;cea\u003c/em\u003e L.) and cabbage (\u003cem\u003eBrassica oler\u0026aacute;cea\u003c/em\u003e L). \u003cem\u003eAgron. Colomb\u003c/em\u003e. \u003cb\u003e38\u003c/b\u003e (2), 205\u0026ndash;217. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.15446/agron.colomb.v38n2.85082\u003c/span\u003e\u003cspan address=\"10.15446/agron.colomb.v38n2.85082\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eStoltz, E. \u0026amp; Nadeau, E. Effects of intercropping on yield, weed incidence, forage quality and soil residual N in organically grown forage maize (\u003cem\u003eZea mays\u003c/em\u003e L.) and faba bean (\u003cem\u003eVicia faba\u003c/em\u003e L). \u003cem\u003eField Crops Res.\u003c/em\u003e \u003cb\u003e169\u003c/b\u003e, 21\u0026ndash;29. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.fcr.2014.09.004\u003c/span\u003e\u003cspan address=\"10.1016/j.fcr.2014.09.004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eShtaya, M. J. Y. et al. Effects of crop mixtures on rust development on faba bean grown in Mediterranean climates. \u003cem\u003eCrop Prot.\u003c/em\u003e \u003cb\u003e146\u003c/b\u003e, 105686. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.cropro.2021.105686\u003c/span\u003e\u003cspan address=\"10.1016/j.cropro.2021.105686\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eStreit, J., Meinen, C., Nelson, W. C. D. \u0026amp; Rauber, R. Above and belowground biomass in a mixed cropping system with eight novel winter faba bean genotypes and winter wheat using FTIR spectroscopy for root species discrimination. \u003cem\u003ePlant. Soil.\u003c/em\u003e \u003cb\u003e36\u003c/b\u003e, 141\u0026ndash;158. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1007/s11104-018-03904-y\u003c/span\u003e\u003cspan address=\"10.1007/s11104-018-03904-y\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eCiampitti, I. A. et al. M. Revisiting Biological Nitrogen Fixation Dynamics in Soybeans. Front. \u003cem\u003ePlant. Sci.\u003c/em\u003e \u003cb\u003e12\u003c/b\u003e, 727021. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fpls.2021.727021\u003c/span\u003e\u003cspan address=\"10.3389/fpls.2021.727021\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eNurgi, N., Tana, T., Dechassa, N., Tesso, B. \u0026amp; Alemayehu, Y. Effect of spatial arrangement of faba bean variety intercropping with maize on yield and yield components of the crops. \u003cem\u003eHeliyon\u003c/em\u003e \u003cb\u003e9\u003c/b\u003e (6), e16751. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.heliyon.2023.e16751\u003c/span\u003e\u003cspan address=\"10.1016/j.heliyon.2023.e16751\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eDzvene, A. R., Tesfuhunei, W. A., Walker, S. \u0026amp; Ceronio, G. Optimizing the planting time and stand density of sunn hemp intercropping for biomass productivity and competitiveness in a maize-based system. \u003cem\u003eField Crops Res\u003c/em\u003e. \u003cb\u003e304\u003c/b\u003e, 109179. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.fcr.2023.109179\u003c/span\u003e\u003cspan address=\"10.1016/j.fcr.2023.109179\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2023).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBybee-Finley, K. A., Mirsky, S. B. \u0026amp; Ryan, M. R. Crop Biomass Not Species Richness Drives Weed Suppression in Warm-Season Annual Grass\u0026ndash;Legume Intercrops in the Northeast. \u003cem\u003eWeed Sci.\u003c/em\u003e \u003cb\u003e65\u003c/b\u003e, 669\u0026ndash;680 (2017). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.jstor.org/stable/26420910\u003c/span\u003e\u003cspan address=\"https://www.jstor.org/stable/26420910\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eBranca, C., Blasi, C. D. \u0026amp; Galgano, A. Experimental analysis about the exploitation of industrial hemp (Cannabis sativa) in pyrolysis, \u003cem\u003eFuel Process. Technol\u003c/em\u003e. 162, 20\u0026ndash;29, (2017). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.fuproc.2017.03.028\u003c/span\u003e\u003cspan address=\"10.1016/j.fuproc.2017.03.028\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eHu, E. et al. Pyrolysis behaviors of corn stover in new two-stage rotary kiln with baffle. \u003cem\u003eJ. Anal. Appl. Pyrol.\u003c/em\u003e \u003cb\u003e161\u003c/b\u003e, 105398. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.jaap.2021.105398\u003c/span\u003e\u003cspan address=\"10.1016/j.jaap.2021.105398\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eAubin, M. P. et al. Industrial hemp response to nitrogen, phosphorus, and potassium fertilization. \u003cem\u003eCrop Forage Turf Man.\u003c/em\u003e \u003cb\u003e1\u003c/b\u003e (1), 1\u0026ndash;10. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.2134/cftm2015.0159\u003c/span\u003e\u003cspan address=\"10.2134/cftm2015.0159\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2016).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eŠarauskis, E. et al. Energy balance, costs and CO2 analysis of tillage technologies in maize cultivation. \u003cem\u003eEnergy\u003c/em\u003e \u003cb\u003e69\u003c/b\u003e, 227\u0026ndash;235. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.energy.2014.02.090\u003c/span\u003e\u003cspan address=\"10.1016/j.energy.2014.02.090\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJasinskas, A. et al. Hemp and Bean Multi-Crop Biomass Pellets and/or Fertilizer. Patent LT6998B. Available online: (2022). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://search.vpb.lt/pdb/patent/dossier/48325/text\u003c/span\u003e\u003cspan address=\"https://search.vpb.lt/pdb/patent/dossier/48325/text\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePetlickaitė, R. et al. Evaluation of the Processing of Multi-Crop Plants into Pelletized Biofuel and Its Use for Energy Conversion, \u003cem\u003eProcesses\u003c/em\u003e. 11(2), 421, (2023). \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/pr11020421\u003c/span\u003e\u003cspan address=\"10.3390/pr11020421\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003ePetlickaitė, R. et al. Evaluation of Multi-Crop Biofuel Pellet Properties and the Life Cycle Assessment. \u003cem\u003eAgric\u003c/em\u003e \u003cb\u003e14\u003c/b\u003e, 1162. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3390/agriculture14071162\u003c/span\u003e\u003cspan address=\"10.3390/agriculture14071162\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2024).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTable 1 to 8 are available in the Supplementary Files section.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"biomass, biofuel pellets, environment, faba bean, maize, multicrops, technical hemp","lastPublishedDoi":"10.21203/rs.3.rs-8209611/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8209611/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHerbaceous plant biomass is an important resource for energy production and an effective strategy to ensure energy supply independence. Growing agricultural plant mixtures (multicrops) contributes to increasing biomass yields, enhancing farm biodiversity, improving soil health, and promoting environmental sustainability.. However, most crop mixtures have not been studied at all. For this reason, research was conducted from 2020 to 2022 at Vytautas Magnus University, Agriculture Academy. The aim of the study was to evaluate the development and productivity of plant mixtures, as well as the related energy and environmental aspects of the applied agrotechnologies, under short-growing-season conditions. Ternary crops tended to be 14% taller, with 24% higher leaf assimilation area, 19% higher chlorophyll index, and 4\u0026ndash;8 times higher first-year dried biomass yields than individual single-species crops. The productivity of the ternary crop reached its highest Comprehensive Evaluation Value (4.54), which was mainly influenced by the chlorophyll index of the leaves. Ternary cultivation was the most fuel-consumptive technology, with 18\u0026ndash;32% higher fuel consumption (103.3 L ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e), due to its higher energy input; however, it generated the most significant net energy (367,668.1 MJ ha\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e) because of its most abundant yield of dried biomass. Ternary crop biomass pellets had the highest density (1,238 kg m\u003csup\u003e-3\u003c/sup\u003e), lower ash content (6%), and the highest ash shrinkage starting temperature (1042\u0026deg; C). It is advisable to cultivate high-capacity yielding ternary crops for one year, which have medium GHG emission and LCA impacts of the pellets produced, but the highest net energy output.\u003c/p\u003e","manuscriptTitle":"Biomass formation and yield performance in diverse multicrops and their potential for biofuel use","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-27 15:49:15","doi":"10.21203/rs.3.rs-8209611/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-03-06T12:49:33+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-03-06T09:12:44+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"107041694593054129403083512543582019301","date":"2026-02-22T16:55:06+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"224715689751555320636060542194216775500","date":"2026-02-21T14:19:07+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"12691612879680364964890839776039990474","date":"2026-01-18T19:49:22+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-01-11T09:17:07+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"87554491713580480032557293632411172447","date":"2026-01-05T07:34:53+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-01-02T14:48:11+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-27T04:54:54+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-27T04:53:01+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-11-26T06:49:13+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"b45f489e-15d0-4188-a09f-b97445961646","owner":[],"postedDate":"November 27th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":58627968,"name":"Biological sciences/Ecology"},{"id":58627969,"name":"Earth and environmental sciences/Ecology"},{"id":58627970,"name":"Earth and environmental sciences/Environmental sciences"},{"id":58627971,"name":"Biological sciences/Plant sciences"}],"tags":[],"updatedAt":"2026-04-07T16:01:32+00:00","versionOfRecord":{"articleIdentity":"rs-8209611","link":"https://doi.org/10.1038/s41598-026-46324-0","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2026-03-30 15:58:25","publishedOnDateReadable":"March 30th, 2026"},"versionCreatedAt":"2025-11-27 15:49:15","video":"","vorDoi":"10.1038/s41598-026-46324-0","vorDoiUrl":"https://doi.org/10.1038/s41598-026-46324-0","workflowStages":[]},"version":"v1","identity":"rs-8209611","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8209611","identity":"rs-8209611","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}