Honey bee pollination improves the yield gaps of Guizotia abyssinica (Asterales: Asteraceae) in Ethiopia

Honey bee pollination improves the yield gaps of Guizotia abyssinica (Asterales: Asteraceae) in Ethiopia | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Honey bee pollination improves the yield gaps of Guizotia abyssinica (Asterales: Asteraceae) in Ethiopia Tolera Kumsa, Tura Bereke, Kasim Roba, Birhanu Mengistu This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4758257/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 5 You are reading this latest preprint version Abstract Niger is an important oilseed crop, and its flowering phenology is highly responsive to cross-pollination. Honey bees (Hymenoptera: Apidae) are the crop's important pollinators due to their adaptation to wide environments. A comparative analysis was performed under field conditions to evaluate the effect of honey bee pollination and fertilizer application on the yield parameters of niger seed. The study was tested under different treatment conditions: (1) plots that were caged by honey bees and received fertilizer; (2) plots that were caged by honey bees but did not receive fertilizer; (3) plots that were restricted from any insect pollination but received fertilizer; and (4) plots that had free access to any pollinators and received fertilizer. The results showed that the mean number of primary branches and capitula per plant were highly significant (P < 0.05) among the treatments. A higher seed yield per ha was obtained in the treatment that received fertilizer and was caged with a honey bee colony (640 kg/ha). The oil content of the niger was higher (35.7%) for the treatment that was caged by honey bees for pollination but did not receive fertilizer. In this study, fertilizer application and honey bee pollination were both important in significantly increasing the seed set and seed yield of the niger plant. However, fertilizer application has less effect on the oil content of niger seed. Therefore, honey bee pollination must be considered in agronomical practices as a factor to sustain the yield and oil content of niger seed. Niger pollination honey bee fertilizer Figures Figure 1 Figure 2 Figure 3 Introduction The niger crop ( Guizotia abyssinica ) is a species in the family Asteraceae that is indigenous to Ethiopia, where it originated, was domesticated, and genetically diversified (Mulatu and Ortiz, 2013 ; Gebeyehu et al., 2022 ). The crop is then spread to India, and other African countries adapt to different environments (Merga, 2022 ). The niger crop, locally named ‘Noug’ is familiar to small-scale farmers and well-integrated into the traditional cropping cycle, grown in rotation with cereals and pulse crops (Getachew, 2019 ; Abebe, 2021 ). The crop contributes about 40% of the total area and 50–60% of oil crop production in Ethiopia (Adane et al., 2021 ; Merga, 2022 ). Nigir crops increase soil fertility and have a weed-depressing effect, and the crop residue left behind after oil extraction is rich in protein and important for livestock feed supplementation (Aleminew et al., 2015 ; Tsehay et al., 2021 ). Niger seed has a high trade value, contributing income to the country through its exports (Tsehay et al., 2021 ). Despite the importance and long cultivation history of the niger crop, it is characterized by a low harvesting index and fertilizer application that promote vegetation growth but are not responsive to increasing seed yield (Aleminew et al., 2015 ; Getachew, 2019 ). Currently, the traditional growing areas of the crop is reduced and is being replaced by high-yielding crops such as hybrid maize and wheat, which makes the crop less competitive (Getinet and Fekadu, 2023 ). Agricultural practices in small-scale farms usually focus on applying fertilizer, irrigation, pesticides and time of plantation as a main factor that affect the yield improvement of different crops (Aleminew et al., 2015 ; Sandeep and Kubsad, 2017 ; Getinet and Fekadu, 2023 ). For pollinator dependent crops this may, however, have limited effect on yield if the availability of effective pollinators is too low (Sawe. et al., 2020). In Ethiopia, of the 76 food crops documented, about 53% are identified as pollinator-dependent (Tibesigwa, 2018 ), indicating that most smallholder farmers are reliant on insect pollination than conventional agricultural inputs (Samnegard et al., 2014 ; Samnegard et al., 2016 ). The flowering phenology of the niger crop is highly responsive to cross-pollination (Sivaram et al., 2013 ), indicating that the plant is heavily reliant on insect pollination to increase its seed yield. However, pollinator populations have experienced serious losses due to human activities, including the use of pesticides and habitat loss that could pose a potential risk to pollinator-dependent crops (Tolera and Ballantyne, 2021 ). Honey bees have a significant advantage over other insect pollinators because they are social and develop a large population that becomes a significant pollinating force. Ethiopia has an estimated 10 million native honeybee colonies (Negash and Greiling, 2017 ), the largest number in Africa. However, little is known about how honey bee pollination affects the yield of the pollinator-dependent crops such as niger crop. Researchers argued that insect pollination enhanced the seed set and yield of pollinator-dependent crops only under higher soil fertility, indicating that limited nutrient availability may affect pollination benefits (Tamburini et al., 2016 ; Garratt et al., 2018 ). This indicated that the synergistic effect between insect pollination and other agronomical factors influences the yields of some crops (Breeze et al., 2014 ; Tamburini et al., 2017 ). Research findings suggest that reducing insect pollination has a two-times greater effect on pollinator-dependent crop yield than a similar reduction in fertilizer application (Fijen et al., 2020 ). However, there is little research evidence that a lack of insect pollination could affect the seed set, seed yield, and quality of the niger. The objective of this study was to evaluate the influence of honey bee pollination and fertilizer application on the yield components of niger crop. Materials and Methods Description of Study Area The study was conducted at the Ginchi research sub-site, Oromiya region, Ethiopia, for two consecutive cropping seasons where the crop is dominantly grown (2021 and 2022). The experimental site is located at 9°01′ N and 38°10′ E, with an altitude of 2300 m above sea level, is characterized by a unimodal rainfall pattern, and receives an average annual rainfall of 1080 mm, mostly from June to September. The annual average temperature is 24°c, and the area is characterized by vertosols. Experimental Setup and Treatments Four treatments were used under the broadcast planting method of niger seed using a randomized complete block design (RCBD), each with three replications. A plot size of 3 m by 3 m was used as an experimental unit. The blocks were separated by 1.5 m of wide-open space, whereas the plots within the blocks were separated by 1 m. The Ginch-niger variety was used as planting material, and seed was broadcasted following the recommended seed rate of 5 kg per ha. Recommended fertilizer (blended N and P) of 5 kg per hectare was applied and evenly scattered during planting time. A 2.5-mm aperture polythene mesh-size cage was prepared and covered on a plot measuring 4 m by 4 m wide and 3 m high for honey bee flight and placed before the start of niger blooming. Treatments were tested to determine the effect of honey bee pollination and fertilizer on the yield and yield components of the niger crop. In the first treatment, plots were received the recommended fertilizer and at the same time caged with a honey bee colony for pollination; in the second treatment, the plots were caged with a honey bee colony for pollination but not received fertilizer; in the third treatment, the plots were received the recommended fertilizer but caged to exclude any insect pollination while allowing the treatment for self and wind pollination; and in the fourth treatment, the plots were received the recommended fertilizer but not caged to be visited by any pollinators. Eight healthy honey bee colonies were prepared for the experiment, and one honey bee colony was placed in a plot assigned for honey bee pollination treatments before the start of niger flowering (Fig. 1 ). All colonies were in single Zander hives, and each was uniformly equalized to have one brood frame with very little pollen in the hive. Sugar syrup (50% water and 50% sugar) was prepared and provided to colonies at intervals of seven days by placing one litter in the feeder, which was placed in the hives to strengthen colonies. Data Collection Niger seed yield is positively correlated with the number of primary branches, number of capitula per plant, number of seeds per capitula, and seed yield/ha (Mulatu and Ortiz, 2013 ). In this study, the number of branches per plant and the number of capitulum (heads) per plant in the plot were counted on ten randomly selected plants before harvesting. The number of mature seeds was counted from ten randomly selected capitula in a plot before harvesting. Seed yield per hectare and thousand dried seeds were randomly taken, counted and weighted. Observations of pollinator visitation as a proxy for pollination were done following the method (Fijen et al., 2018 ). A representative plot of 1 m by 1 m from the treatment outside the cage was randomly selected, and observations were made for different pollinators at four-day intervals throughout the flowering period of the niger crop. Observations were made in the morning from 8:00 a.m. to 11:30 a.m. and in the evening from 13:00 p.m. to 16:30 p.m. each for ten minutes. Samples of pollinators were trapped from the flowers using an insect trap and identified at the Holeta bee research herbarium laboratory. The niger oil content was determined according to the standards in the Holeta Agricultural Research Center oil laboratory. Thirty (30) grams of cleaned and uniform-sized seed were prepared from each treatment. The samples were carefully ground to a uniform size without forming a paste. The flour was sieved using a 1 mm sieve to obtain a uniform particle size. The flour was then extracted with hexane using a soxlet extractor, and hexane was recovered by distillation using a chiller. A solvent was dried at 103°C for 60 minutes at atmospheric pressure. The extracted oil was weighted using an analytical balance with 0.0001 precision. The oil content of seeds was determined by oil/seed (m/m%). The mean of the triplicate was taken as the oil content of the sample calculated as follows: Where m 1 = mass of dried extracted substance in gram m 2 = mass of test portion in gram w = percent of moisture Data analysis We investigated how honey bee pollination and fertilizer affect niger yield parameters by comparing four experimental treatments, each with three replications. Honey bee pollination and fertilizer application were considered as fixed effects and replications as random effects. To analyze and compare the effects of pollination and fertilizer on niger yield parameters, we used one-way ANOVA. When the effects of the treatments were statistically significant, the post hoc range test with standard error of the differences (SED) was used to determine how the treatments differed from each other at an alpha level of 0.05. Statistical analysis was performed using the R-Project software (Version R 4.2.2) and Sigma-Plot version 12.5 was used to draw graphs. Results Yield Parameters of Niger The number of primary branches of the niger plant was significantly different among treatments (F (3, 236) = 49.4, p < 0.05). The highest number of primary branches of the niger plant were recorded in treatments that received fertilizer (8.26 ± 0.14), and the lowest number of primary branches were recorded on plants that were not receiving fertilizer (6.0 ± 0.17). Table 1 . The results showed that the increased number of primary branches might be due to fertilizer applications that promote the vegetation growth of the niger plant. The number of capitula per plant also showed a highly significant difference among treatments (F (3, 236) = 80.9, p < 0.05). The highest capitula number per plant was recorded on plants that received fertilizer (53.9 ± 0.9), compared to the treatment that did not receive fertilizer (35.3 ± 0.8) (Table 1 ). The higher number of branches due to fertilizer application might increase the number of capitula per plant. The mean number of seeds per capitula showed significant differences among treatments (F (3, 236) = 25.6, p < 0.05). Higher seeds were recorded on plants that received fertilizer and were caged with honey bee colonies for pollination (40.1 ± 0.6), compared with the number of seeds that received fertilizer but were excluded from any pollinators (31.6 ± 0.7). The main reason that niger produced a lower number of seeds per capitula might be due to insufficient insect pollination to produce enough seeds. Moreover, in the absence of insect pollination, the treatment exhibited extended flowering for a mean of 12 days compared to the pollinated treatments. Seed Yield per Hectare The niger seed yield per ha was significantly affected by both honey bee pollination and fertilizer application (F (3, 20) = 31.8, p < 0.05). The higher seed yield/ha was recorded in the treatment that received fertilizer and was caged with a honey bee colony for pollination (640.6 ± 11 kg/ha). The lowest seed yield was recorded in the treatment that received fertilizers but was excluded from any pollinators (472.7 ± 9.4 kg/ha); see Table 1 . This might be fertilizer application could result in an increase in the number of branches per plant, which is attributed to an increase in the number of capitula per plant, and after efficient pollination, more seed is produced. Thousand Seeds Weight The same pattern was seen for the thousand seed weight that were affected by treatment, with pollinator-excluded treatment significantly lower than all other treatments (F (3, 20) = 10.4, p < 0.05). The heavier seed was recorded on the treatment that received fertilizer and was honey bee pollinated (4.3 ± 0.13 g), compared to the treatment that received fertilizer but was excluded from any insect pollination (3.48 ± 0.9 g) (Table 1 and Fig. 2 ). Table 1 Niger-yield parameters, mean (± SE) number of branches/plant, number of capitula/plant, number of seeds/ capitula, yield in kg/ha and weight of 1000 seeds for each treatment. Treatments with the same letter are not significantly different (P < 0.05). Treatments Number of main branches/plant Number of capitula/ plant Number of seeds/ capitula Yield in kg/ha 1000 seed weight Fertilizer caged with a honey bee colony 8.2 ± 0.14a 51.3 ± 0.9a 40.1 ± 0.6a 640.6 ± 11a 4.3 ± 0.13a Fertilizer with open pollination 8.5 ± 0.14a 53.9 ± 0.9a 37.6 ± 0.7a 583.5 ± 9.0 b 3.5 ± 0.15b No fertilizer caged with honey bee colony 6.0 ± 0.17b 35.3 ± 0.8b 33.3 ± 0.6b 574.8 ± 6.2c 3.49 ± 0.09b Fertilizer excluded from any insect pollination 8.1 ± 0.15a 51.2 ± 1.0a 31.6 ± 0.7b 472.7 ± 9.4d 3.48 ± 0.9b Oil Content of Niger Seed Inadequate pollination affects not only the yield and seed weight of the niger seed but also the oil content of the seed. The oil content of niger seed was significantly different among treatments (F (3, 8) = 69.5, p < 0.05). The oil content of niger seed varied from 31.6 ± 0.15% for the treatment that received fertilizer but was excluded from any insect pollination to 35.7 ± 0.1% for the treatment that did not receive fertilizer but was caged with honey bees for pollination (Fig. 3 ). Insect Pollinators It was observed that niger flowers were visited by different insect pollinators. During the study, a total of 14 species of pollinators were observed foraging on the flower. Honey bees were the most dominant pollinators, contributing 82.7% of the insect pollinators. Other than honey bees, pollinators included Musca domestica ( 8.72% ), Myathropa florae ( 3.39% ), Eupeodes luniger (1.91 % ), Meliscaeva auricollis (1.25%), and Calliphora vicina (1.19%) (Table 2 ). During the study, it was observed that honey bees were dominantly gathering niger pollen in the morning, starting at 7:00, to 11:30, and then shifting to nectar collection in the afternoon. Table 2 Percentage of insect pollinators visiting niger flower during the study Scientific name Family Percentage proportion Apis mellifera Apidae 82.69 Musca domestica Muscidae 8.72 Myathropa florea Syrphidae 3.39 Eupeodes luniger Syrphidae 1.91 Meliscaeva auricollis Syrphidae 1.25 Calliphoravicina Calliphoridae 1.19 Amata alicia Erebidae 0.3 Lexias pardalis Nymphalidae 0.12 Phalaena euphemia … 0.12 Anthene princeps Lycaenidae 0.06 Cosmetra tumulata Tortricidae 0.06 Epiphora fournierae --- 0.06 Graphomya maculata Muscidae 0.06 Platycorynus dejeani Chrysomelidae 0.06 Discussion The research findings indicated that the synergetic effect of honey bee pollination and fertilizer application provides better seed yield. This may be due to the use of fertilizer, which promotes vegetative growth and increases the number of capitula per branch, while efficient honey bee pollination increases the plant's seed yield. On the other hand, the honey bee pollination treatment provided significant seed yield compared to those excluded from any pollinators, indicating fertilizer application alone had no effect on niger seed yield. Previous studies indicated that deploying honey bee colonies for watermelon pollination resulted in a higher fruit yield (Sawe et al., 2020 ). Our findings are consistent with previous research that indicates insect pollination is more important than other agricultural inputs for pollinator-dependent crops (Tamburini et al., 2017 ; Fijen et al., 2018 ). In our study, applying the required fertilizer by itself cannot compensate for the lack of insect pollination, but the interaction between insect pollination and fertilizer application can improve niger yield. Similar results were found in the cucumber plant, showing that the absence of honey bee pollination reduced the yield by 75% compared to the absence of fertilizer application, which reduced the yield by 18% (Motzke et al., 2015 ). The weight of a thousand seeds was higher in the plots that received fertilizer, and honey bee pollination indicated that insect pollination resulted in a heavier seed weight. Plants receiving enough pollination produced larger, more prolific, and heavier seeds compared to non-pollinated crops (Sagwe et al., 2023 ). Therefore, better utilization of honey bees for niger pollination and strengthening management measures to support other pollinators may be more cost-effective. A higher seed yield was recorded in the open pollination treatment compared to the treatment that excluded pollinators. This might be due to the fact that in open field conditions, pollination might have been accomplished by the interaction of different pollinators that enhanced the seed yield of the niger. Studies shows that pollinator diversity increases the quantity and quality of the crop yield and the yield stability of pollinator dependent crops (Katumo et al., 2022 ). The oil content of the niger seed was higher for the treatment that received no fertilizer and was caged with honey bees for pollination. This showed that honey bee pollination could affect the oil content of the seed. It was suggested that external nitrogen application results in overproduction of protein, causing a reduction in oil content in rapeseed plants (Zapletalová et al., 2022 ). However, in terms of our knowledge, it is difficult to interpret how fertilizer application can negatively influence the oil content of niger seed. Besides, it is evident that honey bee pollination can increase the oil content of sunflower seeds and also affect the composition of fatty acids in the oil, which can enhance its market value (Silva et al., 2018 ). In the present study, it was observed that the treatment excluded from any pollinators exhibited a significantly longer flowering time compared to the pollinated treatments demonstrates the role of insect pollinators in determining flowering length. Other research findings indicated that in the absence of insect pollination, flowers had a longer flowering time (Fung and Thomson, 2017 ; Spigler, 2017 ). Moreover, the adequately pollinated flower have shorter flowering period will likely drive earlier maturation, enabling seed to ripen evenly and allowing for earlier harvest without shattering (Fairhurst et al., 2022 ). However, it needs further study to determine whether the absence of insect pollination could affect the flowering length of the plant, as it is determined by different factors, such as temperature and the availability of water and nutrients (Castro et al., 2008 ). As a compensatory response to the current decline of wild pollinators, crop yield improvements should develop an approach that aims to incorporate honey bee pollination into agronomic practices. During insect pollinators' observations, honey bees are predominantly foraging niger flowers, and the foragers actively collect niger pollen in the morning (7:00–11:30). It is important to take full advantage of managed honey bees in niger pollination because honey bee colonies can draw foraging workers from their colonies and quickly establish dense populations for efficient pollination of the flowers (Samnegård, 2020 ). As the yield advantage of niger depends on insect pollination, honey bee colony management for pollination may be a key factor in the sustainable yield improvement of the crop. Beekeepers may therefore profit from relocating their colonies to niger fields in order to harvest surplus honey, and niger growers are likely to maximize seed yield. Conclusions The present research findings provide a new way of thinking about the need for honey bee pollination for crops that depend on insect pollination. Honey bee pollination affected the yield of niger compared with the exclusion of fertilizer application. As niger is a mass flowering crop and completely outcrossing species with a self-incompatibility nature, it needs sufficient insect pollination through its flowering periods to produce optimum seed yield. Therefore, a substantial need for awareness of honey bee pollination as a crucial factor in increasing the seed yield and oil content of niger. It is necessary to take full advantage of managed honey bees to thrive in niger pollination and compensate for the loss of other native pollinators. Therefore, it is essential to develop the pollination management of honey bees in such a way that it benefits beekeepers through honey production and niger growers through pollination. Declarations Competing interests On behalf of all the authors, I hereby declare that there is no conflict of interest with other people or organizations, which could inappropriately influence this work. Acknowledgments We are grateful to Ethiopian Institute of Agricultural Research (EIAR) and Oromia Agricultural Research Institute (OARI) for their finical and logistic support. Our thanks are also extended to Tadesse Debele, Tesfaye Abera and Konjit Asfaw for their help in experimental layout, field data collection and data entry to computer. References Abebe H (2021) Genetic gain in grain yield and oil content of noug (Guizotia abyssinica) in Ethiopia. J Biosci Agric Res 27(01):2234–2251 Adane G, Hammenhag C, Ortiz R et al (2021) Characterization of oilseed crop noug (Guizotia abyssinica) using agro-morphological traits. Agronomy 11:1–14 Aleminew A, Alemayehu G, Adgo E et al (2015) Response of noug (Guizotia abyssinica Cass.) to Np fertilizers application and seeding rates on yield and yield components in Ebinat District, Amhara region, Ethiopia. World J Agric Sci 11(2):70–83 Breeze TD, Vaissière BE, Bommarco R et al (2014) Agricultural policies exacerbate honeybee pollination service supply-demand mismatches across Europe. PLoS ONE 9(1):e82996 Castro S, Silveira P, Navarro L (2008) Effect of pollination on floral longevity and costs of delaying fertilization in the out-crossing Polygala vayredae Costa (Polygalaceae). Ann Bot 102(6):1043–1048 Fairhurst SM, Jackson GE, Evans A et al (2022) The effect of pollination on the growth and reproduction of oilseed rape (Brassica napus). Basic Appl Ecol 63:164–174 Fijen TPM, Scheper JA, Boom TM et al (2018) Insect pollination is at least as important for marketable crop yield as plant quality in a seed crop. Ecol Lett :1–10 Fijen TPM, Scheper JA, Vogel C et al (2020) Insect pollination is the weakest link in the production of a hybrid seed crop. Agric Ecosyst Environ 290:1–8 Fung HF, Thomson J (2017) Does lack of pollination extend flower life? J Pollinat Ecol 21:86–91 Garratt MPD, Bishop J, Degani E et al (2018) Insect pollination as an agronomic input: Strategies for oilseed rape production. J Appl Ecol 55:2834–2842 Gebeyehu A, Hammenhag C, Tesfaye K et al (2022) RNA-Seq provides novel genomic resources for noug (Guizotia abyssinica) and reveals microsatellite frequency and distribution in its transcriptome. Front Plant Sci 13:1–16 Getachew A (2019) Oilseeds engine for economic development. In: Geremew T, Adugna W, Dereje G, editors. Crop management practices in Noug (Guizotia abyssinica) in the central highlands of Ethiopia. The second oil crops research and development review workshop. Addis Ababa Getinet A, Fekadu G (2023) Towards edible oil self-sufficiency in Ethiopia: Lessons and prospects. Cogent Food Agric 9(1):1–19 Katumo DM, Liang H, Ochola AC et al (2022) Pollinator diversity benefits natural and agricultural ecosystems, environmental health, and human welfare. Plant Divers 44:429–435. https://doi.org/10.1016/j.pld.2022.01.005 Merga M (2022) Niger improvement and production in Ethiopia: progress and major challenges. J Innov Agric 9(1):1 Motzke I, Tscharntke T, Wanger TC et al (2015) Pollination mitigates cucumber yield gaps more than pesticide and fertilizer use in tropical smallholder gardens. J Appl Ecol 52:261–269 Mulatu G, Ortiz R (2013) The importance of Guizotia abyssinica (niger) for sustainable food security in Ethiopia. Genet Resour Crop Evol 60:1763–1770 Negash B, Greiling J (2017) Quality focused apiculture sector value chaindevelopment in Ethiopia. J Agric Sci Technol 7:107–116 Sagwe RN, Peters MK, Dubois T et al (2023) Insect pollination and pollinator supplementation enhances fruit weight, quality, and marketability of avocado (Persea americana). Arthropod Plant Interact 17(6):753–763 Samnegård U (2020) Why is arabica coffee visited by so few non-Apis bees in its native range? Ecology 101(9):1–4 Samnegard U, Hambäck P, Hamba PA et al (2016) A heterogeneous landscape does not guarantee high crop pollination. In: Proceedings of Royal Society B 283. pp. 1–9 Samnegard U, Hambäck PA, Nemomissa S et al (2014) Dominance of the semi-wild honeybee as coffee pollinator across a gradient of shade-tree structure in Ethiopia. J Trop Ecol 30(5):401–408 Sandeep KR, Kubsad VS (2017) Growth and yield of niger (Guizotia abyssinica Cass.) as influenced by planting geometry, fertilizer levels and cycocel spray. J Farm Sci 30(2):172–176 Sawe. T, Eldegard K, Totland Ø et al (2020) Enhancing pollination is more effective than increased conventional agriculture inputs for improving watermelon yields. Ecol Evol 10(12):5343–5353. https://doi.org/10.1002/ece3.6278 Sawe T, Nielsen A, Eldegard K (2020) Crop pollination in small-scale agriculture in Tanzania: household dependence, awareness and conservation. sustainability. 12:su12062228 Silva CAS, Godoy WAC, Jacob CRO et al (2018) Bee pollination highly improves oil quality in sun-flower. Sociobiology 65(4):583–590 Sivaram V, Jayaramappa KV, Menon A et al (2013) Use of bee-attractants in increasing crop productivity in niger (Guizotia abyssinica. L). Brazilian Arch Biol Technol 56(3):365–370 Spigler RB (2017) Plasticity of floral longevity and floral display in the self-compatible biennial Sabatia angularis (Gentianaceae): Untangling the role of multiple components of pollination. Ann Bot 119(1):167–176 Tamburini G, Berti A, Morari F et al (2016) Degradation of soil fertility can cancel pollination benefits in sunflower. Oecologia 180(2):581–587 Tamburini G, Lami F, Marini L (2017) Pollination benefits are maximized at intermediate nutrient levels. In: Proceedings of the Royal Society B: Biological Sciences. Vol. 284. pp. 1–7 Tibesigwa B (2018) Naturally available pollinator decline will decrease household food security and increase the gender gap in nutrition between men and women who head smallholder farm households in Sub-Saharan Africa. In: Environment for development. p. 54 Tolera K, Ballantyne G (2021) Insect pollination and sustainable agriculture in Sub-Saharan Africa. J Pollinat Ecol 27:36–46 Tsehay S, Ortiz R, Geleta M et al (2021) Nutritional profile of the Ethiopian oilseed crop noug (Guizotia abyssinica cass.): Opportunities for its improvement as a source for human nutrition. Foods 10(8):1–15 Zapletalová A, Ducsay L, Varga L et al (2022) Influence of nitrogen nutrition on fatty acids in oilseed rape (Brassica napus L). Plants 11:1–9 Cite Share Download PDF Status: Under Review Version 1 posted Reviewers agreed at journal 04 Oct, 2024 Reviewers invited by journal 01 Aug, 2024 Editor invited by journal 24 Jul, 2024 Editor assigned by journal 24 Jul, 2024 First submitted to journal 22 Jul, 2024 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4758257","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":334655060,"identity":"0f658d6c-2936-4d52-a712-5f7aa59283be","order_by":0,"name":"Tolera Kumsa","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAxUlEQVRIiWNgGAWjYFAC5gbGBiDFz8DARpwGHgZGiBbJBpK1GBwgVos9e2Pjw5lt2+SMbyQ/e/ChgkGeX+wAAVt4DjYbbmy7bWx2I83ccMYZBsOZsxMIaJFIbJN82HY7cduNBDNp3jaGBIPbhLW0/wRqqd88I/0b0VraGIEOSzCQyCHWljMHmyVnnLsN9MabMskZZyQI+4W9vfngx56y2/L87enbJD5U2MjzSxPQggACYJUSxCoHAf4DpKgeBaNgFIyCkQQAmfRFf2xVUEYAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0003-0706-7438","institution":"Oromia Agriculture Research Institute, Holeta Bee Research","correspondingAuthor":true,"prefix":"","firstName":"Tolera","middleName":"","lastName":"Kumsa","suffix":""},{"id":334655061,"identity":"5f0ded07-5926-4da9-8adf-dd63ffa709e4","order_by":1,"name":"Tura Bereke","email":"","orcid":"","institution":"Oromia Agricultural Research Inistitute, Holeta Bee Research","correspondingAuthor":false,"prefix":"","firstName":"Tura","middleName":"","lastName":"Bereke","suffix":""},{"id":334655062,"identity":"14f00b46-7614-483b-8f32-fd2c9a815530","order_by":2,"name":"Kasim Roba","email":"","orcid":"","institution":"Oromia Agricultural Research Institute, Holeta Bee Research","correspondingAuthor":false,"prefix":"","firstName":"Kasim","middleName":"","lastName":"Roba","suffix":""},{"id":334655063,"identity":"3ae1005d-77ae-4a39-9e31-83eff01e7641","order_by":3,"name":"Birhanu Mengistu","email":"","orcid":"","institution":"Ethiopian Institute of Agricultural Research","correspondingAuthor":false,"prefix":"","firstName":"Birhanu","middleName":"","lastName":"Mengistu","suffix":""}],"badges":[],"createdAt":"2024-07-17 18:29:19","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4758257/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4758257/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":63453109,"identity":"c756ec22-c7b6-4753-a0e4-48488b71930a","added_by":"auto","created_at":"2024-08-28 09:44:53","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":316444,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe treatment setup of the experiment with and without honey bee colony\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"image1.png","url":"https://assets-eu.researchsquare.com/files/rs-4758257/v1/d5881312d8d57d87997f4f06.png"},{"id":63453107,"identity":"9273ef2b-13d1-4f0d-9daf-21351f5b07a7","added_by":"auto","created_at":"2024-08-28 09:44:53","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":934268,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eNiger seeds (a) received fertilizer and honey bee pollination (b) niger seed excluded from any insect pollination but received fertilizer\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"image2.png","url":"https://assets-eu.researchsquare.com/files/rs-4758257/v1/f91291a9abab62b1cd69e3a5.png"},{"id":63453108,"identity":"5d9031fe-666c-4c92-b67b-c83ca12cd3ba","added_by":"auto","created_at":"2024-08-28 09:44:53","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":21629,"visible":true,"origin":"","legend":"\u003cp\u003eFigures show the mean percent of oil content of niger seed of different treatments (percent mean ±SE) indicate significant difference among treatments at P \u0026lt; 0.05\u003c/p\u003e","description":"","filename":"image3.png","url":"https://assets-eu.researchsquare.com/files/rs-4758257/v1/f3290be0178c15c21e9322a5.png"},{"id":63453110,"identity":"1cf534ae-2402-4462-9a32-a35b4d8a95cf","added_by":"auto","created_at":"2024-08-28 09:44:58","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1763827,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4758257/v1/be5819c5-f306-4ba4-a0e4-d842a86080ad.pdf"}],"financialInterests":"","formattedTitle":"Honey bee pollination improves the yield gaps of Guizotia abyssinica (Asterales: Asteraceae) in Ethiopia","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe niger crop (\u003cem\u003eGuizotia abyssinica\u003c/em\u003e) is a species in the family \u003cem\u003eAsteraceae\u003c/em\u003e that is indigenous to Ethiopia, where it originated, was domesticated, and genetically diversified (Mulatu and Ortiz, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Gebeyehu et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The crop is then spread to India, and other African countries adapt to different environments (Merga, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). The niger crop, locally named \u0026lsquo;Noug\u0026rsquo; is familiar to small-scale farmers and well-integrated into the traditional cropping cycle, grown in rotation with cereals and pulse crops (Getachew, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Abebe, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The crop contributes about 40% of the total area and 50\u0026ndash;60% of oil crop production in Ethiopia (Adane et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Merga, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Nigir crops increase soil fertility and have a weed-depressing effect, and the crop residue left behind after oil extraction is rich in protein and important for livestock feed supplementation (Aleminew et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Tsehay et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Niger seed has a high trade value, contributing income to the country through its exports (Tsehay et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eDespite the importance and long cultivation history of the niger crop, it is characterized by a low harvesting index and fertilizer application that promote vegetation growth but are not responsive to increasing seed yield (Aleminew et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Getachew, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Currently, the traditional growing areas of the crop is reduced and is being replaced by high-yielding crops such as hybrid maize and wheat, which makes the crop less competitive (Getinet and Fekadu, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Agricultural practices in small-scale farms usually focus on applying fertilizer, irrigation, pesticides and time of plantation as a main factor that affect the yield improvement of different crops (Aleminew et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Sandeep and Kubsad, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Getinet and Fekadu, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). For pollinator dependent crops this may, however, have limited effect on yield if the availability of effective pollinators is too low (Sawe. et al., 2020).\u003c/p\u003e \u003cp\u003eIn Ethiopia, of the 76 food crops documented, about 53% are identified as pollinator-dependent (Tibesigwa, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), indicating that most smallholder farmers are reliant on insect pollination than conventional agricultural inputs (Samnegard et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Samnegard et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). The flowering phenology of the niger crop is highly responsive to cross-pollination (Sivaram et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), indicating that the plant is heavily reliant on insect pollination to increase its seed yield. However, pollinator populations have experienced serious losses due to human activities, including the use of pesticides and habitat loss that could pose a potential risk to pollinator-dependent crops (Tolera and Ballantyne, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Honey bees have a significant advantage over other insect pollinators because they are social and develop a large population that becomes a significant pollinating force. Ethiopia has an estimated 10\u0026nbsp;million native honeybee colonies (Negash and Greiling, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), the largest number in Africa. However, little is known about how honey bee pollination affects the yield of the pollinator-dependent crops such as niger crop.\u003c/p\u003e \u003cp\u003eResearchers argued that insect pollination enhanced the seed set and yield of pollinator-dependent crops only under higher soil fertility, indicating that limited nutrient availability may affect pollination benefits (Tamburini et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Garratt et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). This indicated that the synergistic effect between insect pollination and other agronomical factors influences the yields of some crops (Breeze et al., \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Tamburini et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Research findings suggest that reducing insect pollination has a two-times greater effect on pollinator-dependent crop yield than a similar reduction in fertilizer application (Fijen et al., \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). However, there is little research evidence that a lack of insect pollination could affect the seed set, seed yield, and quality of the niger. The objective of this study was to evaluate the influence of honey bee pollination and fertilizer application on the yield components of niger crop.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eDescription of Study Area\u003c/h2\u003e \u003cp\u003eThe study was conducted at the Ginchi research sub-site, Oromiya region, Ethiopia, for two consecutive cropping seasons where the crop is dominantly grown (2021 and 2022). The experimental site is located at 9\u0026deg;01\u0026prime; N and 38\u0026deg;10\u0026prime; E, with an altitude of 2300 m above sea level, is characterized by a unimodal rainfall pattern, and receives an average annual rainfall of 1080 mm, mostly from June to September. The annual average temperature is 24\u0026deg;c, and the area is characterized by vertosols.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eExperimental Setup and Treatments\u003c/h2\u003e \u003cp\u003eFour treatments were used under the broadcast planting method of niger seed using a randomized complete block design (RCBD), each with three replications. A plot size of 3 m by 3 m was used as an experimental unit. The blocks were separated by 1.5 m of wide-open space, whereas the plots within the blocks were separated by 1 m. The Ginch-niger variety was used as planting material, and seed was broadcasted following the recommended seed rate of 5 kg per ha. Recommended fertilizer (blended N and P) of 5 kg per hectare was applied and evenly scattered during planting time. A 2.5-mm aperture polythene mesh-size cage was prepared and covered on a plot measuring 4 m by 4 m wide and 3 m high for honey bee flight and placed before the start of niger blooming.\u003c/p\u003e \u003cp\u003eTreatments were tested to determine the effect of honey bee pollination and fertilizer on the yield and yield components of the niger crop. In the first treatment, plots were received the recommended fertilizer and at the same time caged with a honey bee colony for pollination; in the second treatment, the plots were caged with a honey bee colony for pollination but not received fertilizer; in the third treatment, the plots were received the recommended fertilizer but caged to exclude any insect pollination while allowing the treatment for self and wind pollination; and in the fourth treatment, the plots were received the recommended fertilizer but not caged to be visited by any pollinators.\u003c/p\u003e \u003cp\u003eEight healthy honey bee colonies were prepared for the experiment, and one honey bee colony was placed in a plot assigned for honey bee pollination treatments before the start of niger flowering (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). All colonies were in single Zander hives, and each was uniformly equalized to have one brood frame with very little pollen in the hive. Sugar syrup (50% water and 50% sugar) was prepared and provided to colonies at intervals of seven days by placing one litter in the feeder, which was placed in the hives to strengthen colonies.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eData Collection\u003c/h2\u003e \u003cp\u003eNiger seed yield is positively correlated with the number of primary branches, number of capitula per plant, number of seeds per capitula, and seed yield/ha (Mulatu and Ortiz, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). In this study, the number of branches per plant and the number of capitulum (heads) per plant in the plot were counted on ten randomly selected plants before harvesting. The number of mature seeds was counted from ten randomly selected capitula in a plot before harvesting. Seed yield per hectare and thousand dried seeds were randomly taken, counted and weighted.\u003c/p\u003e \u003cp\u003eObservations of pollinator visitation as a proxy for pollination were done following the method (Fijen et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). A representative plot of 1 m by 1 m from the treatment outside the cage was randomly selected, and observations were made for different pollinators at four-day intervals throughout the flowering period of the niger crop. Observations were made in the morning from 8:00 a.m. to 11:30 a.m. and in the evening from 13:00 p.m. to 16:30 p.m. each for ten minutes. Samples of pollinators were trapped from the flowers using an insect trap and identified at the Holeta bee research herbarium laboratory.\u003c/p\u003e \u003cp\u003eThe niger oil content was determined according to the standards in the Holeta Agricultural Research Center oil laboratory. Thirty (30) grams of cleaned and uniform-sized seed were prepared from each treatment. The samples were carefully ground to a uniform size without forming a paste. The flour was sieved using a 1 mm sieve to obtain a uniform particle size. The flour was then extracted with hexane using a soxlet extractor, and hexane was recovered by distillation using a chiller. A solvent was dried at 103\u0026deg;C for 60 minutes at atmospheric pressure. The extracted oil was weighted using an analytical balance with 0.0001 precision. The oil content of seeds was determined by oil/seed (m/m%). The mean of the triplicate was taken as the oil content of the sample calculated as follows:\u003c/p\u003e \u003cp\u003e\u003cimg src=\"data:image/png;base64,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\"\u003e\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\u003cp\u003e \u003c/p\u003e \u003cp\u003eWhere\u003c/p\u003e \u003cp\u003em \u003csub\u003e1\u003c/sub\u003e= mass of dried extracted substance in gram\u003c/p\u003e \u003cp\u003em \u003csub\u003e2\u003c/sub\u003e = mass of test portion in gram\u003c/p\u003e \u003cp\u003ew\u0026thinsp;=\u0026thinsp;percent of moisture\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eData analysis\u003c/h2\u003e \u003cp\u003eWe investigated how honey bee pollination and fertilizer affect niger yield parameters by comparing four experimental treatments, each with three replications. Honey bee pollination and fertilizer application were considered as fixed effects and replications as random effects. To analyze and compare the effects of pollination and fertilizer on niger yield parameters, we used one-way ANOVA. When the effects of the treatments were statistically significant, the post hoc range test with standard error of the differences (SED) was used to determine how the treatments differed from each other at an alpha level of 0.05. Statistical analysis was performed using the R-Project software (Version R 4.2.2) and Sigma-Plot version 12.5 was used to draw graphs.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eYield Parameters of Niger\u003c/h2\u003e \u003cp\u003eThe number of primary branches of the niger plant was significantly different among treatments (F \u003csub\u003e(3, 236)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;49.4, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The highest number of primary branches of the niger plant were recorded in treatments that received fertilizer (8.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14), and the lowest number of primary branches were recorded on plants that were not receiving fertilizer (6.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17). Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The results showed that the increased number of primary branches might be due to fertilizer applications that promote the vegetation growth of the niger plant. The number of capitula per plant also showed a highly significant difference among treatments (F \u003csub\u003e(3, 236)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;80.9, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The highest capitula number per plant was recorded on plants that received fertilizer (53.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9), compared to the treatment that did not receive fertilizer (35.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8) (Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The higher number of branches due to fertilizer application might increase the number of capitula per plant.\u003c/p\u003e \u003cp\u003eThe mean number of seeds per capitula showed significant differences among treatments (F \u003csub\u003e(3, 236)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;25.6, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Higher seeds were recorded on plants that received fertilizer and were caged with honey bee colonies for pollination (40.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6), compared with the number of seeds that received fertilizer but were excluded from any pollinators (31.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7). The main reason that niger produced a lower number of seeds per capitula might be due to insufficient insect pollination to produce enough seeds. Moreover, in the absence of insect pollination, the treatment exhibited extended flowering for a mean of 12 days compared to the pollinated treatments.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eSeed Yield per Hectare\u003c/h2\u003e \u003cp\u003eThe niger seed yield per ha was significantly affected by both honey bee pollination and fertilizer application (F \u003csub\u003e(3, 20)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;31.8, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The higher seed yield/ha was recorded in the treatment that received fertilizer and was caged with a honey bee colony for pollination (640.6\u0026thinsp;\u0026plusmn;\u0026thinsp;11 kg/ha). The lowest seed yield was recorded in the treatment that received fertilizers but was excluded from any pollinators (472.7\u0026thinsp;\u0026plusmn;\u0026thinsp;9.4 kg/ha); see Table \u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. This might be fertilizer application could result in an increase in the number of branches per plant, which is attributed to an increase in the number of capitula per plant, and after efficient pollination, more seed is produced.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eThousand Seeds Weight\u003c/h2\u003e \u003cp\u003eThe same pattern was seen for the thousand seed weight that were affected by treatment, with pollinator-excluded treatment significantly lower than all other treatments (F \u003csub\u003e(3, 20)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;10.4, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The heavier seed was recorded on the treatment that received fertilizer and was honey bee pollinated (4.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13 g), compared to the treatment that received fertilizer but was excluded from any insect pollination (3.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9 g) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eNiger-yield parameters, mean (\u0026plusmn;\u0026thinsp;SE) number of branches/plant, number of capitula/plant, number of seeds/ capitula, yield in kg/ha and weight of 1000 seeds for each treatment. Treatments with the same letter are not significantly different (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNumber of main branches/plant\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNumber of capitula/ plant\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNumber of seeds/ capitula\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eYield in kg/ha\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1000 seed weight\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFertilizer caged with a honey bee colony\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e51.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e40.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e640.6\u0026thinsp;\u0026plusmn;\u0026thinsp;11a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.13a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFertilizer with open pollination\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.14a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e53.9\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e37.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e583.5\u0026thinsp;\u0026plusmn;\u0026thinsp;9.0 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo fertilizer caged with honey bee colony\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.17b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e33.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e574.8\u0026thinsp;\u0026plusmn;\u0026thinsp;6.2c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.49\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFertilizer excluded from any insect pollination\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e51.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.0a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e31.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e472.7\u0026thinsp;\u0026plusmn;\u0026thinsp;9.4d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.48\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9b\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eOil Content of Niger Seed\u003c/h2\u003e \u003cp\u003eInadequate pollination affects not only the yield and seed weight of the niger seed but also the oil content of the seed. The oil content of niger seed was significantly different among treatments (F \u003csub\u003e(3, 8)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;69.5, p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The oil content of niger seed varied from 31.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.15% for the treatment that received fertilizer but was excluded from any insect pollination to 35.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1% for the treatment that did not receive fertilizer but was caged with honey bees for pollination (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eInsect Pollinators\u003c/h2\u003e \u003cp\u003eIt was observed that niger flowers were visited by different insect pollinators. During the study, a total of 14 species of pollinators were observed foraging on the flower. Honey bees were the most dominant pollinators, contributing 82.7% of the insect pollinators. Other than honey bees, pollinators included \u003cem\u003eMusca domestica (\u003c/em\u003e8.72%\u003cem\u003e), Myathropa florae (\u003c/em\u003e3.39%\u003cem\u003e), Eupeodes luniger (1.91\u003c/em\u003e%\u003cem\u003e), Meliscaeva auricollis\u003c/em\u003e (1.25%), and \u003cem\u003eCalliphora vicina\u003c/em\u003e (1.19%) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). During the study, it was observed that honey bees were dominantly gathering niger pollen in the morning, starting at 7:00, to 11:30, and then shifting to nectar collection in the afternoon.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePercentage of insect pollinators visiting niger flower during the study\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eScientific name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFamily\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePercentage proportion\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eApis mellifera\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eApidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e82.69\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eMusca domestica\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMuscidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eMyathropa florea\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSyrphidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.39\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eEupeodes luniger\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSyrphidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.91\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eMeliscaeva auricollis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSyrphidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.25\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCalliphoravicina\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCalliphoridae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eAmata alicia\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eErebidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eLexias pardalis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNymphalidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePhalaena euphemia\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026hellip;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eAnthene princeps\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLycaenidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCosmetra tumulata\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTortricidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eEpiphora fournierae\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eGraphomya maculata\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMuscidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePlatycorynus dejeani\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eChrysomelidae\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe research findings indicated that the synergetic effect of honey bee pollination and fertilizer application provides better seed yield. This may be due to the use of fertilizer, which promotes vegetative growth and increases the number of capitula per branch, while efficient honey bee pollination increases the plant's seed yield. On the other hand, the honey bee pollination treatment provided significant seed yield compared to those excluded from any pollinators, indicating fertilizer application alone had no effect on niger seed yield. Previous studies indicated that deploying honey bee colonies for watermelon pollination resulted in a higher fruit yield (Sawe et al., \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Our findings are consistent with previous research that indicates insect pollination is more important than other agricultural inputs for pollinator-dependent crops (Tamburini et al., \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Fijen et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn our study, applying the required fertilizer by itself cannot compensate for the lack of insect pollination, but the interaction between insect pollination and fertilizer application can improve niger yield. Similar results were found in the cucumber plant, showing that the absence of honey bee pollination reduced the yield by 75% compared to the absence of fertilizer application, which reduced the yield by 18% (Motzke et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). The weight of a thousand seeds was higher in the plots that received fertilizer, and honey bee pollination indicated that insect pollination resulted in a heavier seed weight. Plants receiving enough pollination produced larger, more prolific, and heavier seeds compared to non-pollinated crops (Sagwe et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Therefore, better utilization of honey bees for niger pollination and strengthening management measures to support other pollinators may be more cost-effective.\u003c/p\u003e \u003cp\u003eA higher seed yield was recorded in the open pollination treatment compared to the treatment that excluded pollinators. This might be due to the fact that in open field conditions, pollination might have been accomplished by the interaction of different pollinators that enhanced the seed yield of the niger. Studies shows that pollinator diversity increases the quantity and quality of the crop yield and the yield stability of pollinator dependent crops (Katumo et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe oil content of the niger seed was higher for the treatment that received no fertilizer and was caged with honey bees for pollination. This showed that honey bee pollination could affect the oil content of the seed. It was suggested that external nitrogen application results in overproduction of protein, causing a reduction in oil content in rapeseed plants (Zapletalov\u0026aacute; et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, in terms of our knowledge, it is difficult to interpret how fertilizer application can negatively influence the oil content of niger seed. Besides, it is evident that honey bee pollination can increase the oil content of sunflower seeds and also affect the composition of fatty acids in the oil, which can enhance its market value (Silva et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the present study, it was observed that the treatment excluded from any pollinators exhibited a significantly longer flowering time compared to the pollinated treatments demonstrates the role of insect pollinators in determining flowering length. Other research findings indicated that in the absence of insect pollination, flowers had a longer flowering time (Fung and Thomson, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Spigler, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Moreover, the adequately pollinated flower have shorter flowering period will likely drive earlier maturation, enabling seed to ripen evenly and allowing for earlier harvest without shattering (Fairhurst et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). However, it needs further study to determine whether the absence of insect pollination could affect the flowering length of the plant, as it is determined by different factors, such as temperature and the availability of water and nutrients (Castro et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). As a compensatory response to the current decline of wild pollinators, crop yield improvements should develop an approach that aims to incorporate honey bee pollination into agronomic practices.\u003c/p\u003e \u003cp\u003eDuring insect pollinators' observations, honey bees are predominantly foraging niger flowers, and the foragers actively collect niger pollen in the morning (7:00\u0026ndash;11:30). It is important to take full advantage of managed honey bees in niger pollination because honey bee colonies can draw foraging workers from their colonies and quickly establish dense populations for efficient pollination of the flowers (Samneg\u0026aring;rd, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). As the yield advantage of niger depends on insect pollination, honey bee colony management for pollination may be a key factor in the sustainable yield improvement of the crop. Beekeepers may therefore profit from relocating their colonies to niger fields in order to harvest surplus honey, and niger growers are likely to maximize seed yield.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe present research findings provide a new way of thinking about the need for honey bee pollination for crops that depend on insect pollination. Honey bee pollination affected the yield of niger compared with the exclusion of fertilizer application. As niger is a mass flowering crop and completely outcrossing species with a self-incompatibility nature, it needs sufficient insect pollination through its flowering periods to produce optimum seed yield. Therefore, a substantial need for awareness of honey bee pollination as a crucial factor in increasing the seed yield and oil content of niger. It is necessary to take full advantage of managed honey bees to thrive in niger pollination and compensate for the loss of other native pollinators. Therefore, it is essential to develop the pollination management of honey bees in such a way that it benefits beekeepers through honey production and niger growers through pollination.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e \u003ch2\u003eCompeting interests\u003c/h2\u003e \u003cp\u003eOn behalf of all the authors, I hereby declare that there is no conflict of interest with other people or organizations, which could inappropriately influence this work.\u003c/p\u003e \u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e \u003cp\u003eWe are grateful to Ethiopian Institute of Agricultural Research (EIAR) and Oromia Agricultural Research Institute (OARI) for their finical and logistic support. Our thanks are also extended to Tadesse Debele, Tesfaye Abera and Konjit Asfaw for their help in experimental layout, field data collection and data entry to computer.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbebe H (2021) Genetic gain in grain yield and oil content of noug (Guizotia abyssinica) in Ethiopia. J Biosci Agric Res 27(01):2234\u0026ndash;2251\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAdane G, Hammenhag C, Ortiz R et al (2021) Characterization of oilseed crop noug (Guizotia abyssinica) using agro-morphological traits. Agronomy 11:1\u0026ndash;14\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAleminew A, Alemayehu G, Adgo E et al (2015) Response of noug (Guizotia abyssinica Cass.) to Np fertilizers application and seeding rates on yield and yield components in Ebinat District, Amhara region, Ethiopia. World J Agric Sci 11(2):70\u0026ndash;83\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBreeze TD, Vaissi\u0026egrave;re BE, Bommarco R et al (2014) Agricultural policies exacerbate honeybee pollination service supply-demand mismatches across Europe. PLoS ONE 9(1):e82996\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCastro S, Silveira P, Navarro L (2008) Effect of pollination on floral longevity and costs of delaying fertilization in the out-crossing Polygala vayredae Costa (Polygalaceae). Ann Bot 102(6):1043\u0026ndash;1048\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFairhurst SM, Jackson GE, Evans A et al (2022) The effect of pollination on the growth and reproduction of oilseed rape (Brassica napus). Basic Appl Ecol 63:164\u0026ndash;174\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFijen TPM, Scheper JA, Boom TM et al (2018) Insect pollination is at least as important for marketable crop yield as plant quality in a seed crop. Ecol Lett :1\u0026ndash;10\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFijen TPM, Scheper JA, Vogel C et al (2020) Insect pollination is the weakest link in the production of a hybrid seed crop. Agric Ecosyst Environ 290:1\u0026ndash;8\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFung HF, Thomson J (2017) Does lack of pollination extend flower life? J Pollinat Ecol 21:86\u0026ndash;91\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarratt MPD, Bishop J, Degani E et al (2018) Insect pollination as an agronomic input: Strategies for oilseed rape production. J Appl Ecol 55:2834\u0026ndash;2842\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGebeyehu A, Hammenhag C, Tesfaye K et al (2022) RNA-Seq provides novel genomic resources for noug (Guizotia abyssinica) and reveals microsatellite frequency and distribution in its transcriptome. Front Plant Sci 13:1\u0026ndash;16\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGetachew A (2019) Oilseeds engine for economic development. In: Geremew T, Adugna W, Dereje G, editors. Crop management practices in Noug (Guizotia abyssinica) in the central highlands of Ethiopia. The second oil crops research and development review workshop. Addis Ababa\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGetinet A, Fekadu G (2023) Towards edible oil self-sufficiency in Ethiopia: Lessons and prospects. Cogent Food Agric 9(1):1\u0026ndash;19\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKatumo DM, Liang H, Ochola AC et al (2022) Pollinator diversity benefits natural and agricultural ecosystems, environmental health, and human welfare. Plant Divers 44:429\u0026ndash;435. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.pld.2022.01.005\u003c/span\u003e\u003cspan address=\"10.1016/j.pld.2022.01.005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMerga M (2022) Niger improvement and production in Ethiopia: progress and major challenges. J Innov Agric 9(1):1\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMotzke I, Tscharntke T, Wanger TC et al (2015) Pollination mitigates cucumber yield gaps more than pesticide and fertilizer use in tropical smallholder gardens. J Appl Ecol 52:261\u0026ndash;269\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMulatu G, Ortiz R (2013) The importance of Guizotia abyssinica (niger) for sustainable food security in Ethiopia. Genet Resour Crop Evol 60:1763\u0026ndash;1770\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNegash B, Greiling J (2017) Quality focused apiculture sector value chaindevelopment in Ethiopia. J Agric Sci Technol 7:107\u0026ndash;116\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSagwe RN, Peters MK, Dubois T et al (2023) Insect pollination and pollinator supplementation enhances fruit weight, quality, and marketability of avocado (Persea americana). Arthropod Plant Interact 17(6):753\u0026ndash;763\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSamneg\u0026aring;rd U (2020) Why is arabica coffee visited by so few non-Apis bees in its native range? Ecology 101(9):1\u0026ndash;4\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSamnegard U, Hamb\u0026auml;ck P, Hamba PA et al (2016) A heterogeneous landscape does not guarantee high crop pollination. In: Proceedings of Royal Society B 283. pp. 1\u0026ndash;9\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSamnegard U, Hamb\u0026auml;ck PA, Nemomissa S et al (2014) Dominance of the semi-wild honeybee as coffee pollinator across a gradient of shade-tree structure in Ethiopia. J Trop Ecol 30(5):401\u0026ndash;408\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSandeep KR, Kubsad VS (2017) Growth and yield of niger (Guizotia abyssinica Cass.) as influenced by planting geometry, fertilizer levels and cycocel spray. J Farm Sci 30(2):172\u0026ndash;176\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSawe. T, Eldegard K, Totland \u0026Oslash; et al (2020) Enhancing pollination is more effective than increased conventional agriculture inputs for improving watermelon yields. Ecol Evol 10(12):5343\u0026ndash;5353. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/ece3.6278\u003c/span\u003e\u003cspan address=\"10.1002/ece3.6278\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSawe T, Nielsen A, Eldegard K (2020) Crop pollination in small-scale agriculture in Tanzania: household dependence, awareness and conservation. sustainability. 12:su12062228\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSilva CAS, Godoy WAC, Jacob CRO et al (2018) Bee pollination highly improves oil quality in sun-flower. Sociobiology 65(4):583\u0026ndash;590\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSivaram V, Jayaramappa KV, Menon A et al (2013) Use of bee-attractants in increasing crop productivity in niger (Guizotia abyssinica. L). Brazilian Arch Biol Technol 56(3):365\u0026ndash;370\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSpigler RB (2017) Plasticity of floral longevity and floral display in the self-compatible biennial Sabatia angularis (Gentianaceae): Untangling the role of multiple components of pollination. Ann Bot 119(1):167\u0026ndash;176\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTamburini G, Berti A, Morari F et al (2016) Degradation of soil fertility can cancel pollination benefits in sunflower. Oecologia 180(2):581\u0026ndash;587\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTamburini G, Lami F, Marini L (2017) Pollination benefits are maximized at intermediate nutrient levels. In: Proceedings of the Royal Society B: Biological Sciences. Vol. 284. pp. 1\u0026ndash;7\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTibesigwa B (2018) Naturally available pollinator decline will decrease household food security and increase the gender gap in nutrition between men and women who head smallholder farm households in Sub-Saharan Africa. In: Environment for development. p. 54\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTolera K, Ballantyne G (2021) Insect pollination and sustainable agriculture in Sub-Saharan Africa. J Pollinat Ecol 27:36\u0026ndash;46\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTsehay S, Ortiz R, Geleta M et al (2021) Nutritional profile of the Ethiopian oilseed crop noug (Guizotia abyssinica cass.): Opportunities for its improvement as a source for human nutrition. Foods 10(8):1\u0026ndash;15\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZapletalov\u0026aacute; A, Ducsay L, Varga L et al (2022) Influence of nitrogen nutrition on fatty acids in oilseed rape (Brassica napus L). Plants 11:1\u0026ndash;9\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"international-journal-of-tropical-insect-science","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jtis","sideBox":"Learn more about [International Journal of Tropical Insect Science](http://link.springer.com/journal/42690)","snPcode":"42690","submissionUrl":"https://www.editorialmanager.com/jtis/default2.aspx","title":"International Journal of Tropical Insect Science","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Niger, pollination, honey bee, fertilizer","lastPublishedDoi":"10.21203/rs.3.rs-4758257/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4758257/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eNiger is an important oilseed crop, and its flowering phenology is highly responsive to cross-pollination. Honey bees (Hymenoptera: Apidae) are the crop's important pollinators due to their adaptation to wide environments. A comparative analysis was performed under field conditions to evaluate the effect of honey bee pollination and fertilizer application on the yield parameters of niger seed. The study was tested under different treatment conditions: (1) plots that were caged by honey bees and received fertilizer; (2) plots that were caged by honey bees but did not receive fertilizer; (3) plots that were restricted from any insect pollination but received fertilizer; and (4) plots that had free access to any pollinators and received fertilizer. The results showed that the mean number of primary branches and capitula per plant were highly significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05) among the treatments. A higher seed yield per ha was obtained in the treatment that received fertilizer and was caged with a honey bee colony (640 kg/ha). The oil content of the niger was higher (35.7%) for the treatment that was caged by honey bees for pollination but did not receive fertilizer. In this study, fertilizer application and honey bee pollination were both important in significantly increasing the seed set and seed yield of the niger plant. However, fertilizer application has less effect on the oil content of niger seed. Therefore, honey bee pollination must be considered in agronomical practices as a factor to sustain the yield and oil content of niger seed.\u003c/p\u003e","manuscriptTitle":"Honey bee pollination improves the yield gaps of Guizotia abyssinica (Asterales: Asteraceae) in Ethiopia","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-28 09:44:48","doi":"10.21203/rs.3.rs-4758257/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2024-10-04T08:36:51+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-08-01T09:15:48+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"International Journal of Tropical Insect Science","date":"2024-07-24T15:28:37+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-07-24T11:05:18+00:00","index":"","fulltext":""},{"type":"submitted","content":"International Journal of Tropical Insect Science","date":"2024-07-23T03:20:39+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"international-journal-of-tropical-insect-science","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jtis","sideBox":"Learn more about [International Journal of Tropical Insect Science](http://link.springer.com/journal/42690)","snPcode":"42690","submissionUrl":"https://www.editorialmanager.com/jtis/default2.aspx","title":"International Journal of Tropical Insect Science","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"e109b36e-7059-4de7-a104-6c7dac172496","owner":[],"postedDate":"August 28th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2024-08-28T09:44:48+00:00","versionOfRecord":[],"versionCreatedAt":"2024-08-28 09:44:48","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4758257","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4758257","identity":"rs-4758257","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}