The comparative assessment of Trichoderma and insecticides on chewing and phloem-feeding herbivores and physiological traits in cucumber

The comparative assessment of Trichoderma and insecticides on chewing and phloem-feeding herbivores and physiological traits in cucumber | 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 The comparative assessment of Trichoderma and insecticides on chewing and phloem-feeding herbivores and physiological traits in cucumber Kamrul Hassan, Salma Sarker, Md Musfiqur Alam, Tonmoy Sarkar, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8410264/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Cucumbers ( Cucumis sativa L ) are economically important vegetable crops globally. However, it is highly susceptible to aphids, whiteflies and red pumpkin beetle that cause significant damage. Currently, chemical pesticide is practiced controlling herbivores, but it has severe health hazards. Trichoderma consider as an eco-friendly management approach for controlling these herbivores. Hence, our study aims to compare the effectiveness of two Trichoderma spp. and insecticides in controlling aphids, whiteflies and red pumpkin beetle of cucumber. We performed semi-field pot experiment with a cucumber variety (F1 hybrid cucumber) using two Trichoderma species (BP1- Trichoderma asperellum (0R125623) @ 2 ´ 10 6 CFU/gm) and BP2- Trichoderma harzianum 2 ´ 10 6 CFU/gm) @ 5gm/liter water), two insecticides (Cord 10 % EC and Cartap 50SP) including a control. Our results found that total insect community were higher in chemical intervention than Trichoderma , showing a significant reduction of herbivorous insect. Trichoderma harzianum showed higher efficacy for controlling herbivores pest density and percent infestation. The foliar application of Trichoderma has positive effects on parasitoids. The incidence of cucumber mosaic virus (%) reduced to 8-10 % than control in dual application of Trichoderma spp. suggested synergistic effects between species. Although total chlorophyll and carotenoids content in leaves remain unaffected, however, the average trichomes per leaves, fruit length and yield/plant significantly increased with both Trichoderma and insecticidal treatments where Trichoderma spp. found effective than insecticide treatment. Our research has broader implications for the management of cucurbits insects and diseases with less dependency on insecticides while considering Trichoderma as a potential alternative. Insect community Biopesticide Cucumber Insecticides Trichoderma asperellum Trichoderma harzianum Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction Cucumber ( Cucumis sativus L. ) is an important vegetable which is widely cultivated across the world (Kaur and Sharma, 2022 ; Malepszy, 1988 ). Cucumbers are attacked by many arthropod pests such as fruit flies, aphids, whiteflies, red pumpkin beetle, two spotted spider mites including diseases such as cucumber mosaic virus transmitted by aphids (Van Steenis et al. 1995; Mauck et al. 2015 ; Leite et al. 2006 ; Part et al. 2002). While there are effective methods to control fruit flies using pheromone traps (Shaha and Chandran, 2023), the control of aphids, whiteflies and red pumpkin beetle heavily depends on conventional pesticides (Kaleem Ullah et al. 2023 ). The application of pesticides negatively effects the biodiversity of natural enemies such as parasitoids and predators while causing pest resistance and secondary pest outbreaks (Bommarco et al. 2011 ; Theiling and Croft, 1988 ). The application of biopesticides products is popular in many advanced countries (Senthil-Nathan, 2014 ) but in Bangladesh the use of biopesticides is limited due to complete dependency on pesticides to manage insects and diseases in cucumber. Hence, application of biopesticides is required for sustainable pest management in cucumber. However, there are not such study conducted that estimates the comparative efficacy of commercially available biopesticides especially Trichoderma products with insecticides on the reduction of aphids, whiteflies and red pumpkin beetles including common mosaic virus transmitted by aphids in cucumber and increasing yield components. Biopesticides are composed with living organism used as alternative to conventional pesticides due its specificity to target organism, less toxicity for non-target organism such as natural enemies and faster decomposition ability in the environment (Liu et al. 2021 ; Hanif et al. 2022 ; Senthil-Nathan, 2014 ). The common biopesticides such as Trichoderma spp. Beauveria bassiana , Bacillus thuringiensis , and nuclear polyhedrosis viruses (NPV) used to control insect pest in vegetables crops, however, their efficacy has not been tested with conventional insecticides in vegetables crops (Fenibo et al. 2022 ; Verma et al. 2024 ). Although, biopesticides are getting popular day by day, but there are some limitations that could impede its application to control insect pest of vegetables. The effects of biopesticides are limited due its slow persistent in the field, to narrow or too broad for pests and slow knock down effects including economic constraints (Essiedu et al. 2020 ; Zaki et al. 2020 ; Fenibo et al. 2021 ; Chapple et al. 1996 ). Hence, in this study, we manipulated the application of Trichoderma spp. single and dual application to enhance its efficacy for controlling three key insects of cucumber and vector borne disease- cucumber mosaic virus transmitted by aphids than conventional approach. Trichoderma is a potential biopesticides as well as biofertilizer, however, its application and its effects on aphids, whiteflies and red pumpkin beetles has not been tested in cucumber. Among Trichoderma spp. Trichoderma harzianum and Trichoderma asperellum is widely used as naturally occurring filamentous fungus which is very effective to control insects and diseases (Ding et al. 2025 ). Trichoderma invade the host body through penetrating cuticle using mechanical and enzymatic action that allow them to grow inside and parasitize the host (Monte, 2023 ; Parrilli et al. 2019 ; Poveda et al. 2021). They also produce secondary chemical compounds which act as antifeedant such as chitinases against rice moth (Vijayakumar et al. 2016 ), linoleate and linoleic acid against aphids (Kaushik et al. 2020 ), and repellent against beetle Acanthoscelides obtectus major stored pest in common bean (Alınç et al. 2021 ; Rodríguez-Gonz´alez et al. 2018a ,). Previous reports also documented that T. harzianum produced secondary metabolites such as phenols and flavonoids that showed strong defense related activity against lepidopteran insects (Dwisandi et al. 2024 ). Moreover, T. harzianum and T. asperellum may produce volatiles compounds that attracted predator and parasitoids as well as causing disruption of insect symbiotic relationship with microbes (Poveda et al. 2021). Due to their high potentiality to control insect pests, the efficacy of both T. harzianum and T. asperellum to control aphids, whiteflies and red pumpkin beetles in cucumber need to study in experimental set-up in compare with insecticides. Therefore, our objectives of this study to investigate the role of two T. harzianum and T. asperellum on the reduction of aphids, whiteflies and red pumpkin beetle population in comparison with conventional insecticidal methods. We performed a semi-field pot experiment with cucumber using two Trichoderma species and two insecticidal treatments with control. We hypothesize that- a) the population of aphids, whiteflies and red pumpkin beetles and percent incidence of cucumber mosaic viruses will be reduced significantly with Trichoderma spp. than conventional treatments, b) We also assume that foliar application of Trichoderma spores may increase the trichome number and density than insecticides and control treatments, c) Foliar spray of Trichoderma may increase chlorophyll and total carotenoids than insecticidal and control treatments due to its potentiality to increase leaf green tissues and d) Foliar spray of Trichoderma may increase fruit size and shape, and yield of cucumber than insecticides treatments due to its potentiality to reduce insect attack through making leaves difficult to feed or less palatable for insects via increasing trichomes numbers and producing antifeedant and repellent chemical compounds. 2. Materials and Method 2.1 Plant species, experimental design and pot preparation The F1 hybrid cucumber Thai Green was used in this study that purchased from Ispahani Agro Limited. The fruit of this species is deep in color and average weight 280–300 gm. This species is widely cultivated by farmers in Bangladesh. Our study composed with an outdoor semi-field pot experiment at Department of Entomology, Sylhet Agricultural University, Bangladesh. The study period was November 2024 to March 2025. The experiment consists of two biopesticides- Trichoderma harzianum and Trichoderma asperellum , two insecticides- contact and systematic and no intervention-spray tap water with 5 replicates that produced 30 experimental units. The size of the pot was 12 inches in height, and 14 inch in width. The total volume of the pot with soil was approximately 12L. The experimental soils were collected from local nursery and kept 7 days before fill into the pots. Prior to fill-up the pots, experimental soils mixed with cow dung (10%), with addition of essential nutrients such as nitrogen, phosphorus and potassium. Each pots received approximately 30 gm N, 10 gm P and 12 gm K, respectively prior to transplanting cucumber seedlings and 30 days after transplanting. The cucumber seedlings of the experiments previously grew in seed trays in sterile potting mixtures. Two weeks old seedling were transplanted in early November 2024. Each pot has two seedlings and kept those until the end of the experiment. Any seedling deaths were recorded during the establishment of the study and replaced when needed. All the pots with transplanted seedlings were kept in outside throughout the study period. The plants were kept outside to record the natural infestations of insect pests and disease occurrence of cucumber. The staking was done with bamboo and iron wire for climbing the plants for proper growth and fruiting. 2.2 Experimental treatments Our study tested two biopesticides- BP1- Trichoderma asperellum (strain 0R125623 -2 × 10 6 CFU/gm) @ 5gm/liter water and BP2- Trichoderma harzianum (2 × 10 6 CFU/gm) @ 5gm/liter water. BP1 + BP2 also applied as dual on the cucumber plants to observe any synergistic effects. We used two insecticides: C1-contact insecticides; Cord 10% EC @ 1ml/liter water and C1 + S1- contact and systematic insecticides; Cartap 50SP @ 2gm/liter water. The treatments were applied directly into the plants after preparation freshly every 15 days interval. The dual application of both Trichoderma spp. assumes to help control both insects and disease of crops. The single and dual mode of action contact (Cord 10% EC) and contact and systematic insecticides (Cord 10% EC + Cartap 50SP) also likely to control both external and internal insect pests of cucumber. Hence single and dual application of Trichoderma and multiple mode of action insecticides may provide the pathway to control wide range of insect pests rather than control specific pest which is often practices in the farmer fields. We also have no intervention (NI) treatment where spray tap water. 2.3 Insect, disease sampling and yield attributes recording Two weeks after transplanting, we started counting total insect community through direct observation but also through sweeping net when required. The number of aphid/plant, whitefly/plant, and red pumpkin beetle/plant were collected through direct observation and sweeping nets. The number of parasitoid/plant were recorded through sweeping. The % infested leaves/plant and mosaic leaves/plant were also collected through direct observation. Total insect/plant (aphids, whiteflies, red pumpkin beetles, other beetles, flies, and many other known and unknown insects), % infested leaves/plant (caterpillar, beetles and other unknown insect damage) and disease incidence (% mosaic leaves only) data were presented after sum of all samplings with treatment combination. The insect and disease incidence data were collected until final harvesting of the cucumber. The average fruit length (cm) and yield (kg) of cucumber was collected after harvesting mature fruits every seven days intervals. The average fruit length (cm) and yield (kg) of cucumber was calculated after adding all samplings and presented/plant. 2.4 Counting of trichomes Three fresh cucumber leaves collected directly from leaves and put under microscope for counting trichomes number. Under the microscope, ten random location was selected and number of trichomes were recorded. We had 60 plants, thereby, fresh leaves were collected every day for counting and summed number of trichomes/leaves. 2.5 Chlorophyll extraction and determination process Approximately 3–4 fresh cucumber leaves were collected directly from the potted plants and immediately cut into small pieces. Approximately 0.25 g fresh leaves were collected from mixed sample and dipped into 10 ml of absolute alcohol in 15 ml falcon tubes and kept in the dark for 24 hours. The extracted chlorophyll was put into the centrifuge for 10 mins at 4000 rpm. After that, the Chlorophyll A, B and total carotenoids were determined with respective wavelengths (665 nm, 649 nm and 470 nm, respectively) using double row spectrophotometer (Model: WA-60D, S/N: 27202305033, WAVE ANALYTICS, D-82362 Weilheim, Germany). The collected data has been calculated using an internal standard curve that was previously prepared with absolute alcohol. The total Chlorophyll was estimated through sum-up Chlorophyll A, and Chlorophyll B and the ratio by dividing Chlorophyll A by Chlorophyll B, respectively. 2.6 Statistical analysis All data analysis were done with R version 4.1.5 (2025-06-13). All data were checked for normality and homogeneity before analysis to meet the assumption of data analysis criteria. The count data such as total insects/plant, aphids/plant, whiteflies/plant, red pumpkin beetles/plant, parasitoids/plant and number of trichomes/leaf were analyzed by General Linear Model using glm function with Poisson family distribution with log-link function (Salinas Ruíz et al. 2023 ). The data such as % infested leaves/plant, mosaic/leaves, total chlorophyll and carotenoids (mg/g fresh leaves), yield and average fruit length were analyzed with Linear Model using glm function from Car packages in R (Fox and Weisberg, 2019). Pairwise comparisons were done among the treatment combinations after using Tukey methods for comparing a family of six estimates using emmeans pakages from R (Length et al. 2018). 3. Results 3.1 Effects of Trichoderma and insecticides on the abundance and diversity of insect communities and % disease incidence Total insect population observed higher in chemical intervention than Trichoderma spp. (χ² = 105.18, df = 5, P < 0.0001 ). The lowest number of insect population found in BP2 treatment (Fig. 1 a). Aphid population significantly changed (χ² = 103.70, df = 5, P < 0.0001 ) with treatments. The higher number of aphids found in both CI & CI + SI treatments than BP1, BP2 or BP1 + BP2 treatments, respectively (Fig. 1 b). BP2 was found the most efficient to control aphid population. In whiteflies and red pumpkin beetles, there were significant effects for treatments (χ² = 67.723, df = 5, P < 0.0001 ; χ² = 59.00, df = 5, P < 0.0001 ) where lowest whiteflies and red pumpkin beetle populations found in BP2 (Fig. 1 c-d). The overall effects of treatments on parasitoids also significant (χ² = 6.038, df = 5, P < 0.001), where the population of parasitoid decreased in insecticides treatments than Trichoderma spp. (Fig. 1 e). The percent infested leaves (by caterpillar, beetles and unknown insects) not significantly affected by treatments, however, lowest infestation documented in BP1 + BP2 treatment (Fig. 1 f), respectively. In disease incidence, the % cucumber mosaic leaves were significantly changed (F 5,24 = 8.536, P < 0.0001) with Trichoderma spp. where lowest mosaic leaves found in BP1 + BP2 than control and insecticides treatments (Fig. 1 g). 3.2 Effects of Trichoderma and insecticides on average number of trichomes/leaf and total chlorophyll and carotenoids content (mg/g fresh weight) The density of leaf trichomes in cucumber plants varied noticeably among treatments. Overall, both Trichoderma spp. and chemical insecticides significantly increased trichome numbers compared to untreated plants (χ² = 50.84, df = 5, P < 0.0001). Interestingly, the highest density was recorded under the combined application of Trichoderma spp. which significantly outperformed the single application and insecticide treatments (Fig. 2 ). Both Trichoderma spp. and synthetic chemical intervention were found non-significant (F 5,12 = 1.823, P = 0.182) in relation to no intervention for total chlorophyll (Fig. 3 a) and carotenoids content (F 5,12 = 1.675, P = 0.214; Fig. 3 b). 3.3 Effects of Trichoderma and insecticides on average fruit length/plant and yield/plant No main effects were found significant between Trichoderma or chemical interventions and no interventions on average fruit length (F 5,24 = 2.203, P = 0.08; Fig. 4 a). The overall yield of cucumber was significantly increased with Trichoderma and insecticides treatments than control (F 5,24 = 2.203, P = 0.08; F 5,24 = 2.514, P < 0.05; Fig. 4 b). Though no significant differences between Trichoderma and insecticides were found but the highest fruit length was observed in T. harzianum & CI + SI treatment (Fig. 4 a) and highest yield was observed in T. asperellum & CI treatment, respectively (Fig. 4 b). 4. Discussion Our findings in this study demonstrate that both Trichoderma spp. are effective enough to control insect pests of cucumber while offering additional ecological benefits in contrast to conventional insecticides that documented in recent studies (Kubiak et al. 2023 ; Monte, 2023 ; Poveda, 2021 ). The dual application of both Trichoderma species not only suppressed tested insects; aphids, whiteflies, and red pumpkin beetle, but also enhanced trichome density, indicating a potential synergistic effect on plant defence mechanisms, and ultimately maintained a minimum number of diseased infested leaves (Kim, 2019 ). Cucumber production around the world suffers considerable losses due to common sap-feeding pests such as aphids ( Aphis gossypii ), whiteflies ( Bemisia tabaci ), and red pumpkin beetle ( Aulacophora foveicollis Lucas). However, the potential and sustainable management approach for these insect pests remain unexplored especially the role of Trichoderma spp . in compare with conventional method. In this study, we documented that in parallel with conventional pesticides, the foliar application of Trichoderma can also potentially minimize the attacks of aphids, whiteflies and red pumpkin beetle in cucumber. Aphid infestations are particularly damaging, as both nymphs and adults suck into the phloem, leading to leaf curling, discoloration, and in severe situations, wilting of entire plants, which translates into reduced fruit yield (Kumari et al. 2021 ). Their sugary secretions, known as honeydew, foster the development of sooty mold, further interfering with photosynthesis and overall plant vitality (Razmjou et al. 2011 ; Kumari et al. 2021 ). Beyond direct injury, aphids also spread destructive viruses including Cucumber mosaic virus (CMV) in cucumber compounding yield losses through disease outbreaks. Our study demonstrated that the reduction of aphid number significantly reduced CMV diseases than insecticides and no intervention treatments suggested that aphids are highly resistant against insecticides due to their ability to produce genetically modified offspring that combat insecticides (Kaleem Ullah et al. 2023 ; Edwards et al. 2008 ). In this study, both Trichoderma spp. was successful to control aphids after foliar application assuming their direct effects on aphids through penetrating soft aphid body by puncturing cuticle and parasitize the host (Coppola et al. 2008; Battaglia et al. 2024 ; Poveda, 2021 ). Our study suggested that Trichoderma spp. is a strong candidate to control cucumber aphids and CMV diseases that transmitted by aphids which is not successfully control by conventional approaches. In addition, Trichoderma spp. may attract the natural enemies such parasitic wasp through releasing volatiles compounds that could also the reduced aphid population in Trichoderma treated plants (Woo et al. 2023 ; Poveda, 2021 ). The ladybird beetle is the key predator to control aphids and has been found very effective in many studies (Omkar & Srivastava, 2023; Riddick, 2017 ; Ceryngier et al. 2012 ). If the ladybird beetle can be attracted by volatiles that released by Trichoderma there likely an indirect plant defence to control aphids and CMV diseases, respectively. On the other hands, the conventional approach killed the target organism including non-target organisms such as ladybird beetle due to their wider mode of action (Bommarco et al. 2011 ; Bielza, 2016 ). A future study hence require undertaking to explore the role of Trichoderma spp. to attract natural enemies especially parasitic wasp or ladybird beetles to control aphids as indirect plant defence in field condition (Poveda, 2021 ). Whiteflies are also key pests in cucumber including many vegetables and fruit crops. They damage cucumber plants through depleting sap, which stunts growth and reduces photosynthetic activity, while also serving as carriers of viral diseases such as leaf curl virus in Tomato (Devendran et al. 2023). In this study, we found that the number of whiteflies significantly reduced with T. harzianum that is slightly higher in insecticidal treatments. Interestingly, the effects of T. asperellum on lowering whiteflies population remain unaffected that suggested species specific effects of Trichoderma on hemipteran insect pest (Fig. 1 ). Our results indicated that we should be careful about the species-specific effects of Trichoderma spp . before its application in field condition. However, the multiple application of Trichoderma spp. could be able to minimize the limitation of species specificity of Trichoderma through controlling wide range of insect pests and is associated plant diseases. While dual application of Trichoderma spp. seems potential but it is important to keep in mind about the synergistic and antagonistic effects of the species within (Poveda, 2021 ). Red pumpkin beetles are also key pests for cucurbits production especially their gregarious feeding in young cucurbits seedlings (Hassan et al. 2012 ). Cucumber and sweet gourds are the most suitable host for red pumpkin beetles at young and mature stage of plants, but they could feed on wide range of cucurbits host (Hassan et al. 2012 ; Saljoqi et al. 2007). The beetles could defoliate the young leaves through chewing within a day and retard the growth of plants. The high infestation of red pumpkin beetles could cause yield loss up to 100% especially at seedling stage if there is no intervention take place (Regmi et al. 2020). The conventional contact insecticides are widely used to control the beetle, but it seems they became insecticides resistant over time (Hassan et al. 2011 ). The alternative approach Trichoderma spp. could be an option to minimize the pest damage but never been tested. Our study however, found that the population dynamics of beetles were significantly reduced with T. harzianum suggested that the foliar application could increase cucumber trichomes density, antifeedant and repellent compounds, that could reduce the feeding of beetle population than control treatments in cucumber (Poveda, 2021 ). The reduction of beetle population was consistent for both Trichoderma spp. thereby, indicated that Trichoderma likely to be an alternative option to manage red pumpkin beetle infestation, however field validation with large cucumber cultivar needed for validate the outcomes. Trichomes are hair-like outgrowths that occur on nearly all non-aquatic plant parts, including leaves, stems, and even fruits (Goffreda et al. 1988 ). They may be glandular or non-glandular, unicellular or multicellular, and often function as a protective barrier against insects and fungal pathogens (Sarria et al. 2010 ). Our study found that dual application of Trichoderma spp. increases the number of trichome suggested to enhance plant protection and self-defense mechanism against sap feeding and chewing herbivores (Fig. 2 ). In addition to direct effects of Trichoderma spp. on sap feeding and chewing herbivore, the increased number of trichome than control treatments suggested an additional benefits for plants that helps to combat insects and pathogens in cucumber. The number of trichomes also increased in insecticidal treatments than control however, our study not able provide explanation about the increase of trichome number in those treatments (Fig. 2 ). Our assumption is that plant respond to increase trichome number when additional crop protection strategy applied to control insect and pathogen attack. The increase of trichome number could negatively influence the feeding of sap feeding insects and chewing beetles, thereby the population of sap feeding and chewing insects reduced in this study with Trichoderma . Previous study documents that trichomes can release sticky or viscous secretions that trap sap feeding insects or deter chewing herbivory by making feeding more difficult (Wagner, 1991 ; Wheeler and Krimmel, 2015 ). Some study also found that insects caught in these secretions may attract predators of herbivores, thereby strengthening the plant’s indirect defense system (Krimmel and Pearse, 2013 ). Non-glandular trichomes, on the other hand, are often rigid or hooked structures that can physically puncture insect bodies or obstruct their movement, directly interfering with feeding behavior (Levin, 1973; Riddick and Simmons, 2014 ). Hooked non-glandular types are particularly effective, as they can ensnare not only a wide range of herbivores but also their natural enemies (Riddick and Simmons, 2014 ). Such entrapments can reduce the survival and reproductive success of insects and may even influence the long-term population dynamics and evolutionary fitness of the species involved (Peterson et al. 2016 ; Riddick and Simmons, 2014 ). Our results also found that chlorophyll (primary photosynthetic pigment); and carotenoid content remain unaffected with treatment combination although we assume that there should be an increase in green tissues with Trichoderma due to its positive effects on leaf parameters that documented in recent study (Paraginski et al. 2025 ). However, there are significant increase of average fruit length and yield of cucumber than control treatments that suggested the positive effects of Trichoderma spp. on the production of cucumber. Although, there are similar results also found in insecticidal treatments, but cucumber produced in biopesticide treatments likely to prefer by consumer than insecticidal treatments due its less or no chemical residues. In nutshell, our research suggested the application of Trichoderma spp . could an alternative option to fight insects and diseases than insecticides to grow sustainable and safe cucurbits for the consumer. 5. Conclusion Our results concluded that both T. asperellum and T. harzianum were reduce the sap feeding and chewing herbivore population in cucumber than insecticidal and control treatments. T. harzianum are likely to be more successful to control sap feeding and chewing herbivore including no negative effects on parasitoids population. The dual application of Trichoderma spp. provided significant outcome through reducing the incidence of CMV diseases that suggested synergistic effects of Trichoderma spp. The effects of insecticides treatment also found significant than control treatment, however, both treatments showed negative effects of parasitoids that suggested its effects on non-target organisms impede natural parasitism of sap feeding insects such as aphids. The increased of trichome number suggested additional benefits of Trichoderma spp. to control pest density by reducing their feeding or repelling them from the leaves, however, more study need to conduct to explore this mechanism. Although our study did not find any positive effects of Trichoderma spp. on photosynthetic tissues but future study focusing on the effects of Trichoderma on leaf green tissues require to tease apart the mechanism. Although the yield parameters of cucumber positively affected by Trichoderma and insecticidal treatments than control, but the negative effects of pesticides could be possible to minimize though using biopesticides as alternative option to control insects and disease of cucumber. Our study has boarder application on the management of cucurbits insects and diseases though considering biopesticides such as Trichoderma as potential option while reducing the application of pesticides for safe and sustainable vegetables production for consumers. Declarations Author contributions KH— conceived, conceptualized, writing—original draft preparation, writing—review and editing, conduct experiments, data curation, analyzed the data, and prepared the figures. MMA- writing- original draft, review and editing. TS—writing—review and editing, and data curation. MFM—writing—review and editing, approved the final draft. SS- writing—review and editing, approved the final draft. MP—writing—review and editing, approved the final draft. Data availability All data will be available upon reasonable request to corresponding author. Funding This research has been funded by UGC-SAURES authority from Sylhet Agricultural University. The gran number is SAURES AG-09. Author declarations The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in the paper. Conflict of interest The authors declare no competing interests. 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Rev.(ECR) , 3 , 16-20. http://doi.org/10.26480/ecr.01.2020.16.20 Riddick, E. W. (2017). Identification of conditions for successful aphid control by ladybirds in greenhouses. Insects , 8 (2), 38. https://doi.org/10.3390/insects8020038 Riddick, E. W. & Simmons, A. M. (2014). Do plant trichomes cause more harm than good to predatory insects? In Pest Management Science (Vol. 70, Issue 11, pp. 1655–1665). John Wiley and Sons Ltd. https://doi.org/10.1002/ps.3772 Rodríguez-Gonz´alez, ´A. Casquero, P.A. Su´arez-Villanueva, V. Carro-Huerga, G. ´Alvarez-García, S. Mayo-Prieto, S. et al. 2018a. Effect of trichodiene production by Trichoderma harzianum on Acanthoscelides obtectus . J. Stored Prod. Res. 77, 231–239. https://doi.org/10.1016/j.jspr.2018.05.001. Saha, T. & Chandran, N. (2023). Evaluation of optimum heights of pheromone trap on attraction of fruit flies (Diptera: Tephritidae) in cucumber in North eastern India. International Journal of Tropical Insect Science , 43 (1), 289-295. https://doi.org/10.1007/s42690-023-00942-7 Saljoqi, A. U. R. & Khan, S. (2007). Relative abundance of the red pumpkin beetle, Aulacophora foveicophora Lucas, on different cucurbitaceous vegetables. Sarhad Journal of Agriculture , 23 (1), 135. Salinas Ruíz, J. Montesinos López, O. A. Hernández Ramírez, G. & Crossa Hiriart, J. (2023). Generalized linear mixed models with applications in agriculture and biology (p. 427). Springer Nature. Sarria, E. Palomares‐Rius, F. J. López‐Sesé, A. I. Heredia, A. & Gómez‐Guillamón, M. L. (2010). Role of leaf glandular trichomes of melon plants in deterrence of Aphis gossypii Glover. Plant Biology , 12 (3), 503-511. https://doi.org/10.1111/j.1438-8677.2009.00239.x Senthil-Nathan, S. (2014). A review of biopesticides and their mode of action against insect pests. Environmental sustainability: Role of green technologies , 49-63. https://doi.org/10.1007/978-81-322-2056-5_3 Theiling, K. M. & Croft, B. A. (1988). Pesticide side-effects on arthropod natural enemies: a database summary. Agriculture, Ecosystems & Environment , 21 (3-4), 191-218. https://doi.org/10.1016/0167-8809(88)90088-6 Van Steenis, M. J. & El‐Khawass, K. A. M. H. (1995). Life history of Aphis gossypii on cucumber: influence of temperature, host plant and parasitism. Entomologia Experimentalis et Applicata , 76 (2), 121-131. https://doi.org/10.1111/j.1570-7458.1995.tb01954.x Verma, M. L. Kumar, A. Chintagunta, A. D. Samudrala, P. J. K. Bardin, M. & Lichtfouse, E. (2024). Microbial production of biopesticides for sustainable agriculture. Sustainability , 16 (17), 7496. https://doi.org/10.3390/su16177496 Vijayakumar, N. Alagar, S. Madanagopal, N. 2016. Effects of chitinase from Trichoderma viride on feeding, growth and biochemical parameters of the rice moth, Corcyra cephalonica Stainton. Journalof Entomology and Zoology. Studies 4, 520–523. Wagner, G. J. (1991). Secreting Glandular Trichomes: More than Just Hairs Plant physiology , 96 (3), 675–679. https://doi.org/10.1104/pp.96.3.675 Wheeler, A. G. & Krimmel, B. A. (2015). Mirid (Hemiptera: Heteroptera) specialists of sticky plants: Adaptations, interactions, and ecological implications. In Annual Review of Entomology (Vol. 60, pp. 393–414). Annual Reviews Inc. https://doi.org/10.1146/annurev-ento-010814-020932 Woo, S.L. Hermosa, R. Lorito, M. et al. Trichoderma : a multipurpose, plant-beneficial microorganism for eco-sustainable agriculture. Nat Rev Microbiol 21 , 312–326 (2023). https://doi.org/10.1038/s41579-022-00819-5 Zaki, O. Weekers, F. Thonart, P. Tesch, E. Kuenemann, P. & Jacques, P. (2020). Limiting factors of mycopesticide development. Biological Control , 144 , 104220. https://doi.org/10.1016/j.biocontrol.2020.104220 Additional Declarations No competing interests reported. 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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-8410264","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":581656623,"identity":"42080790-0252-4537-8b00-99692e36095a","order_by":0,"name":"Kamrul Hassan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA8UlEQVRIiWNgGAWjYBACCQkUbgWUZmxgJkYLSNEZkrUwthGhRXJ28+OPP2ruMPBL9x98dHNenbzBAeaHHxh3WOPUIi1zzEya59gzBsk5h5mNc7cdNtxwgM1YgvFMOk4tchIJZswMbIcZDG4ks0nnbjvAuOEAgxnQhYfxaEn//PHHv8MM9mAtc+rsNxxg/4ZXi7REjoEEL1CBgQRISwNz4oYDPPhtkZyRUybN23eYR+JGsrFxzrHDyTMP8xRLJOLxi8SN9M0ff3w7LMc/I/Hh45yaOtu+4+0bP3zEE2IwwINggmIkgaCGUTAKRsEoGAX4AADzoVGK6eLjLgAAAABJRU5ErkJggg==","orcid":"","institution":"Sylhet Agricultural University","correspondingAuthor":true,"prefix":"","firstName":"Kamrul","middleName":"","lastName":"Hassan","suffix":""},{"id":581656624,"identity":"78b2177b-f768-4419-8aef-714a9bd328d5","order_by":1,"name":"Salma Sarker","email":"","orcid":"","institution":"Australian National University","correspondingAuthor":false,"prefix":"","firstName":"Salma","middleName":"","lastName":"Sarker","suffix":""},{"id":581656625,"identity":"b8ad8878-2701-4ac7-bf7a-178e7a378a06","order_by":2,"name":"Md Musfiqur Alam","email":"","orcid":"","institution":"Sylhet Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Md","middleName":"Musfiqur","lastName":"Alam","suffix":""},{"id":581656626,"identity":"c56ca401-ca29-41cf-8b95-c4924b2dd214","order_by":3,"name":"Tonmoy Sarkar","email":"","orcid":"","institution":"Sylhet Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Tonmoy","middleName":"","lastName":"Sarkar","suffix":""},{"id":581656627,"identity":"b312c173-8c1f-4dd1-a4c0-7c704d70b9b5","order_by":4,"name":"Md Fuad Mondal","email":"","orcid":"","institution":"Sylhet Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Md","middleName":"Fuad","lastName":"Mondal","suffix":""},{"id":581656628,"identity":"630731d5-e05b-43f5-9a5c-b238f9768ad8","order_by":5,"name":"Mahfuza Pervin","email":"","orcid":"","institution":"Western Sydney University","correspondingAuthor":false,"prefix":"","firstName":"Mahfuza","middleName":"","lastName":"Pervin","suffix":""}],"badges":[],"createdAt":"2025-12-20 07:23:54","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8410264/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8410264/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":101444292,"identity":"872a6a0e-dc05-44d1-af7b-4617f841d68b","added_by":"auto","created_at":"2026-01-29 18:01:28","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":495181,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of \u003cem\u003eTrichoderma\u003c/em\u003e(single and dual) and insecticides (contact and contact + systematic) application on- a) Total insect/plant, b) Aphids/plant, c) Whiteflies/leaf, d) Red pumpkin beetle/plant, e) Parasitoids/plant, f) Infested leaves (%)/plant, and g) Mosaic leaves/plant. Chi-square with corresponding P-values originated from GLM output (a-e) and F-values with corresponding P-values originated from LM outputs (f-g). The treatments are NI; No intervention, BP1; \u003cem\u003eTrichoderma asperellum\u003c/em\u003e, BP1; \u003cem\u003eTrichoderma harzianum\u003c/em\u003e, BP1+BP2; both species applied, CI; Cord 10 % EC, CI+SI; Cord 10 % EC + Cartap 50SP. The detailed information of treatments found in methods section.\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8410264/v1/dd2146e7671e0bd103203c97.jpeg"},{"id":101444291,"identity":"94206ed7-bbe7-45f5-a7e4-eda0b9fb1887","added_by":"auto","created_at":"2026-01-29 18:01:28","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":112638,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of \u003cem\u003eTrichoderma\u003c/em\u003e (single and dual) and insecticides (contact and contact + systematic) application on the number of trichomes densities per leaf in cucumber. Chi-square and P-values originated from GLM output. Different lower-case letter indicated significant differences among the treatment combinations after using Tukey methods for comparing a family of six estimates using emmeans pakages from R. The treatments are NI; No intervention, BP1; \u003cem\u003eTrichoderma asperellum\u003c/em\u003e, BP1; \u003cem\u003eTrichoderma harzianum\u003c/em\u003e, BP1+BP2; both species, CI; Cord 10 % EC, CI+SI; Cord 10 % EC + Cartap 50SP. The detailed information of treatments found in methods section.\u003c/p\u003e\n\u003cp\u003eBoth \u003cem\u003eTrichoderma spp.\u003c/em\u003e and synthetic chemical intervention were found non-significant (F\u003csub\u003e5,12\u003c/sub\u003e= 1.823, P = 0.182) in relation to no intervention for total chlorophyll (Fig. 3a) and carotenoids content (F\u003csub\u003e5,12\u003c/sub\u003e= 1.675, P = 0.214; Fig. 3b).\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-8410264/v1/84adb37d52089297d5231edc.png"},{"id":101444289,"identity":"79436fbf-8cf7-4763-8cb2-2f81ca5d7d99","added_by":"auto","created_at":"2026-01-29 18:01:28","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":163535,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of \u003cem\u003eTrichoderma\u003c/em\u003e(single and dual) and insecticides (contact and contact + systematic) application on chlorophyll (3a) and carotenoids (3b) content (mg/g fresh weight). F statistics and corresponding P-values originated from Linear Model (LM) output. The treatments are NI; No intervention, BP1; \u003cem\u003eTrichoderma asperellum\u003c/em\u003e, BP1; \u003cem\u003eTrichoderma harzianum\u003c/em\u003e, BP1+BP2; both species applied, CI; Cord 10 % EC, CI+SI; Cord 10 % EC + Cartap 50SP. The detailed information of treatments found in methods section.\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-8410264/v1/ba9f9dbd734d47780bb1719e.png"},{"id":101444290,"identity":"807a7240-eda2-4d64-98c7-b785b9b35de6","added_by":"auto","created_at":"2026-01-29 18:01:28","extension":"jpeg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":242662,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of \u003cem\u003eTrichoderma\u003c/em\u003e (single and dual) and insecticides (contact and contact + systematic) application on average fruit length (4a) and yield (4b) on cucumber. F statistics and corresponding P-values originated from Linear Model (LM) output. Different lower-case letter indicated significant differences among the treatment combinations after using Tukey methods for comparing a family of six estimates using emmeans pakages from R. The treatments are NI; No intervention, BP1; \u003cem\u003eTrichoderma asperellum\u003c/em\u003e, BP1; \u003cem\u003eTrichoderma harzianum\u003c/em\u003e, BP1+BP2; both species applied, CI; Cord 10 % EC, CI+SI; Cord 10 % EC + Cartap 50SP. The detailed information of treatments found in methods section.\u003c/p\u003e","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8410264/v1/b17c12d409feb01b728efa09.jpeg"},{"id":101880527,"identity":"c826a728-b648-4560-bd62-35773bd5275f","added_by":"auto","created_at":"2026-02-04 15:03:14","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1796758,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8410264/v1/b8f36d8c-03d9-48d2-a63b-f4bd9ab2754c.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"The comparative assessment of Trichoderma and insecticides on chewing and phloem-feeding herbivores and physiological traits in cucumber","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eCucumber (\u003cem\u003eCucumis sativus L.\u003c/em\u003e) is an important vegetable which is widely cultivated across the world (Kaur and Sharma, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Malepszy, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1988\u003c/span\u003e). Cucumbers are attacked by many arthropod pests such as fruit flies, aphids, whiteflies, red pumpkin beetle, two spotted spider mites including diseases such as cucumber mosaic virus transmitted by aphids (Van Steenis et al. 1995; Mauck et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Leite et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Part et al. 2002). While there are effective methods to control fruit flies using pheromone traps (Shaha and Chandran, 2023), the control of aphids, whiteflies and red pumpkin beetle heavily depends on conventional pesticides (Kaleem Ullah et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The application of pesticides negatively effects the biodiversity of natural enemies such as parasitoids and predators while causing pest resistance and secondary pest outbreaks (Bommarco et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Theiling and Croft, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e1988\u003c/span\u003e). The application of biopesticides products is popular in many advanced countries (Senthil-Nathan, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) but in Bangladesh the use of biopesticides is limited due to complete dependency on pesticides to manage insects and diseases in cucumber. Hence, application of biopesticides is required for sustainable pest management in cucumber. However, there are not such study conducted that estimates the comparative efficacy of commercially available biopesticides especially \u003cem\u003eTrichoderma\u003c/em\u003e products with insecticides on the reduction of aphids, whiteflies and red pumpkin beetles including common mosaic virus transmitted by aphids in cucumber and increasing yield components.\u003c/p\u003e \u003cp\u003eBiopesticides are composed with living organism used as alternative to conventional pesticides due its specificity to target organism, less toxicity for non-target organism such as natural enemies and faster decomposition ability in the environment (Liu et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Hanif et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Senthil-Nathan, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The common biopesticides such as \u003cem\u003eTrichoderma spp. Beauveria bassiana\u003c/em\u003e, \u003cem\u003eBacillus thuringiensis\u003c/em\u003e, and nuclear polyhedrosis viruses (NPV) used to control insect pest in vegetables crops, however, their efficacy has not been tested with conventional insecticides in vegetables crops (Fenibo et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Verma et al. \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Although, biopesticides are getting popular day by day, but there are some limitations that could impede its application to control insect pest of vegetables. The effects of biopesticides are limited due its slow persistent in the field, to narrow or too broad for pests and slow knock down effects including economic constraints (Essiedu et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Zaki et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Fenibo et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Chapple et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1996\u003c/span\u003e). Hence, in this study, we manipulated the application of \u003cem\u003eTrichoderma spp.\u003c/em\u003e single and dual application to enhance its efficacy for controlling three key insects of cucumber and vector borne disease- cucumber mosaic virus transmitted by aphids than conventional approach.\u003c/p\u003e \u003cp\u003e \u003cem\u003eTrichoderma\u003c/em\u003e is a potential biopesticides as well as biofertilizer, however, its application and its effects on aphids, whiteflies and red pumpkin beetles has not been tested in cucumber. Among \u003cem\u003eTrichoderma spp. Trichoderma harzianum\u003c/em\u003e and \u003cem\u003eTrichoderma asperellum\u003c/em\u003e is widely used as naturally occurring filamentous fungus which is very effective to control insects and diseases (Ding et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). \u003cem\u003eTrichoderma\u003c/em\u003e invade the host body through penetrating cuticle using mechanical and enzymatic action that allow them to grow inside and parasitize the host (Monte, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Parrilli et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Poveda et al. 2021). They also produce secondary chemical compounds which act as antifeedant such as chitinases against rice moth (Vijayakumar et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), linoleate and linoleic acid against aphids (Kaushik et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), and repellent against beetle \u003cem\u003eAcanthoscelides obtectus\u003c/em\u003e major stored pest in common bean (Alın\u0026ccedil; et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Rodr\u0026iacute;guez-Gonz\u0026acute;alez et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2018a\u003c/span\u003e,). Previous reports also documented that \u003cem\u003eT. harzianum\u003c/em\u003e produced secondary metabolites such as phenols and flavonoids that showed strong defense related activity against lepidopteran insects (Dwisandi et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2024\u003c/span\u003e). Moreover, \u003cem\u003eT. harzianum\u003c/em\u003e and \u003cem\u003eT. asperellum\u003c/em\u003e may produce volatiles compounds that attracted predator and parasitoids as well as causing disruption of insect symbiotic relationship with microbes (Poveda et al. 2021). Due to their high potentiality to control insect pests, the efficacy of both \u003cem\u003eT. harzianum\u003c/em\u003e and \u003cem\u003eT. asperellum\u003c/em\u003e to control aphids, whiteflies and red pumpkin beetles in cucumber need to study in experimental set-up in compare with insecticides.\u003c/p\u003e \u003cp\u003eTherefore, our objectives of this study to investigate the role of two \u003cem\u003eT. harzianum\u003c/em\u003e and \u003cem\u003eT. asperellum\u003c/em\u003e on the reduction of aphids, whiteflies and red pumpkin beetle population in comparison with conventional insecticidal methods. We performed a semi-field pot experiment with cucumber using two \u003cem\u003eTrichoderma species\u003c/em\u003e and two insecticidal treatments with control. We hypothesize that- a) the population of aphids, whiteflies and red pumpkin beetles and percent incidence of cucumber mosaic viruses will be reduced significantly with \u003cem\u003eTrichoderma spp.\u003c/em\u003e than conventional treatments, b) We also assume that foliar application of \u003cem\u003eTrichoderma\u003c/em\u003e spores may increase the trichome number and density than insecticides and control treatments, c) Foliar spray of \u003cem\u003eTrichoderma\u003c/em\u003e may increase chlorophyll and total carotenoids than insecticidal and control treatments due to its potentiality to increase leaf green tissues and d) Foliar spray of \u003cem\u003eTrichoderma\u003c/em\u003e may increase fruit size and shape, and yield of cucumber than insecticides treatments due to its potentiality to reduce insect attack through making leaves difficult to feed or less palatable for insects via increasing trichomes numbers and producing antifeedant and repellent chemical compounds.\u003c/p\u003e"},{"header":"2. Materials and Method","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1 Plant species, experimental design and pot preparation\u003c/h2\u003e \u003cp\u003eThe F1 hybrid cucumber Thai Green was used in this study that purchased from Ispahani Agro Limited. The fruit of this species is deep in color and average weight 280\u0026ndash;300 gm. This species is widely cultivated by farmers in Bangladesh. Our study composed with an outdoor semi-field pot experiment at Department of Entomology, Sylhet Agricultural University, Bangladesh. The study period was November 2024 to March 2025. The experiment consists of two biopesticides- \u003cem\u003eTrichoderma harzianum\u003c/em\u003e and \u003cem\u003eTrichoderma asperellum\u003c/em\u003e, two insecticides- contact and systematic and no intervention-spray tap water with 5 replicates that produced 30 experimental units. The size of the pot was 12 inches in height, and 14 inch in width. The total volume of the pot with soil was approximately 12L. The experimental soils were collected from local nursery and kept 7 days before fill into the pots. Prior to fill-up the pots, experimental soils mixed with cow dung (10%), with addition of essential nutrients such as nitrogen, phosphorus and potassium. Each pots received approximately 30 gm N, 10 gm P and 12 gm K, respectively prior to transplanting cucumber seedlings and 30 days after transplanting. The cucumber seedlings of the experiments previously grew in seed trays in sterile potting mixtures. Two weeks old seedling were transplanted in early November 2024. Each pot has two seedlings and kept those until the end of the experiment. Any seedling deaths were recorded during the establishment of the study and replaced when needed. All the pots with transplanted seedlings were kept in outside throughout the study period. The plants were kept outside to record the natural infestations of insect pests and disease occurrence of cucumber. The staking was done with bamboo and iron wire for climbing the plants for proper growth and fruiting.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Experimental treatments\u003c/h2\u003e \u003cp\u003eOur study tested two biopesticides- BP1- \u003cem\u003eTrichoderma asperellum (strain 0R125623\u003c/em\u003e -2 \u0026times; 10\u003csup\u003e6\u003c/sup\u003e CFU/gm) @ 5gm/liter water and BP2-\u003cem\u003eTrichoderma harzianum\u003c/em\u003e (2 \u0026times; 10\u003csup\u003e6\u003c/sup\u003e CFU/gm) @ 5gm/liter water. BP1\u0026thinsp;+\u0026thinsp;BP2 also applied as dual on the cucumber plants to observe any synergistic effects. We used two insecticides: C1-contact insecticides; Cord 10% EC @ 1ml/liter water and C1\u0026thinsp;+\u0026thinsp;S1- contact and systematic insecticides; Cartap 50SP @ 2gm/liter water. The treatments were applied directly into the plants after preparation freshly every 15 days interval. The dual application of both \u003cem\u003eTrichoderma spp.\u003c/em\u003e assumes to help control both insects and disease of crops. The single and dual mode of action contact (Cord 10% EC) and contact and systematic insecticides (Cord 10% EC\u0026thinsp;+\u0026thinsp;Cartap 50SP) also likely to control both external and internal insect pests of cucumber. Hence single and dual application of \u003cem\u003eTrichoderma\u003c/em\u003e and multiple mode of action insecticides may provide the pathway to control wide range of insect pests rather than control specific pest which is often practices in the farmer fields. We also have no intervention (NI) treatment where spray tap water.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Insect, disease sampling and yield attributes recording\u003c/h2\u003e \u003cp\u003eTwo weeks after transplanting, we started counting total insect community through direct observation but also through sweeping net when required. The number of aphid/plant, whitefly/plant, and red pumpkin beetle/plant were collected through direct observation and sweeping nets. The number of parasitoid/plant were recorded through sweeping. The % infested leaves/plant and mosaic leaves/plant were also collected through direct observation. Total insect/plant (aphids, whiteflies, red pumpkin beetles, other beetles, flies, and many other known and unknown insects), % infested leaves/plant (caterpillar, beetles and other unknown insect damage) and disease incidence (% mosaic leaves only) data were presented after sum of all samplings with treatment combination. The insect and disease incidence data were collected until final harvesting of the cucumber. The average fruit length (cm) and yield (kg) of cucumber was collected after harvesting mature fruits every seven days intervals. The average fruit length (cm) and yield (kg) of cucumber was calculated after adding all samplings and presented/plant.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Counting of trichomes\u003c/h2\u003e \u003cp\u003eThree fresh cucumber leaves collected directly from leaves and put under microscope for counting trichomes number. Under the microscope, ten random location was selected and number of trichomes were recorded. We had 60 plants, thereby, fresh leaves were collected every day for counting and summed number of trichomes/leaves.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Chlorophyll extraction and determination process\u003c/h2\u003e \u003cp\u003eApproximately 3\u0026ndash;4 fresh cucumber leaves were collected directly from the potted plants and immediately cut into small pieces. Approximately 0.25 g fresh leaves were collected from mixed sample and dipped into 10 ml of absolute alcohol in 15 ml falcon tubes and kept in the dark for 24 hours. The extracted chlorophyll was put into the centrifuge for 10 mins at 4000 rpm. After that, the Chlorophyll A, B and total carotenoids were determined with respective wavelengths (665 nm, 649 nm and 470 nm, respectively) using double row spectrophotometer (Model: WA-60D, S/N: 27202305033, WAVE ANALYTICS, D-82362 Weilheim, Germany). The collected data has been calculated using an internal standard curve that was previously prepared with absolute alcohol. The total Chlorophyll was estimated through sum-up Chlorophyll A, and Chlorophyll B and the ratio by dividing Chlorophyll A by Chlorophyll B, respectively.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Statistical analysis\u003c/h2\u003e \u003cp\u003eAll data analysis were done with R version 4.1.5 (2025-06-13). All data were checked for normality and homogeneity before analysis to meet the assumption of data analysis criteria. The count data such as total insects/plant, aphids/plant, whiteflies/plant, red pumpkin beetles/plant, parasitoids/plant and number of trichomes/leaf were analyzed by General Linear Model using glm function with Poisson family distribution with log-link function (Salinas Ru\u0026iacute;z et al. \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). The data such as % infested leaves/plant, mosaic/leaves, total chlorophyll and carotenoids (mg/g fresh leaves), yield and average fruit length were analyzed with Linear Model using glm function from Car packages in R (Fox and Weisberg, 2019). Pairwise comparisons were done among the treatment combinations after using Tukey methods for comparing a family of six estimates using emmeans pakages from R (Length et al. 2018).\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cp\u003e \u003cb\u003e3.1 Effects of\u003c/b\u003e \u003cb\u003eTrichoderma\u003c/b\u003e \u003cb\u003eand insecticides on the abundance and diversity of insect communities and % disease incidence\u003c/b\u003e\u003c/p\u003e \u003cp\u003eTotal insect population observed higher in chemical intervention than \u003cem\u003eTrichoderma\u003c/em\u003e spp. (χ\u0026sup2; = 105.18, df\u0026thinsp;=\u0026thinsp;5, \u003cem\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.0001\u003c/em\u003e). The lowest number of insect population found in BP2 treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ea). Aphid population significantly changed (χ\u0026sup2; = 103.70, df\u0026thinsp;=\u0026thinsp;5, \u003cem\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.0001\u003c/em\u003e) with treatments. The higher number of aphids found in both CI \u0026amp; CI\u0026thinsp;+\u0026thinsp;SI treatments than BP1, BP2 or BP1\u0026thinsp;+\u0026thinsp;BP2 treatments, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eb). BP2 was found the most efficient to control aphid population. In whiteflies and red pumpkin beetles, there were significant effects for treatments (χ\u0026sup2; = 67.723, df\u0026thinsp;=\u0026thinsp;5, \u003cem\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.0001\u003c/em\u003e; χ\u0026sup2; = 59.00, df\u0026thinsp;=\u0026thinsp;5, \u003cem\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.0001\u003c/em\u003e) where lowest whiteflies and red pumpkin beetle populations found in BP2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ec-d). The overall effects of treatments on parasitoids also significant (χ\u0026sup2; = 6.038, df\u0026thinsp;=\u0026thinsp;5, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001), where the population of parasitoid decreased in insecticides treatments than \u003cem\u003eTrichoderma spp.\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ee). The percent infested leaves (by caterpillar, beetles and unknown insects) not significantly affected by treatments, however, lowest infestation documented in BP1\u0026thinsp;+\u0026thinsp;BP2 treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003ef), respectively. In disease incidence, the % cucumber mosaic leaves were significantly changed (F\u003csub\u003e5,24\u003c/sub\u003e = 8.536, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001) with \u003cem\u003eTrichoderma spp.\u003c/em\u003e where lowest mosaic leaves found in BP1\u0026thinsp;+\u0026thinsp;BP2 than control and insecticides treatments (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eg).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e3.2 Effects of\u003c/b\u003e \u003cb\u003eTrichoderma\u003c/b\u003e \u003cb\u003eand insecticides on average number of trichomes/leaf and total chlorophyll and carotenoids content (mg/g fresh weight)\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe density of leaf trichomes in cucumber plants varied noticeably among treatments. Overall, both \u003cem\u003eTrichoderma spp.\u003c/em\u003e and chemical insecticides significantly increased trichome numbers compared to untreated plants (χ\u0026sup2; = 50.84, df\u0026thinsp;=\u0026thinsp;5, P\u0026thinsp;\u0026lt;\u0026thinsp;0.0001). Interestingly, the highest density was recorded under the combined application of \u003cem\u003eTrichoderma spp.\u003c/em\u003e which significantly outperformed the single application and insecticide treatments (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eBoth \u003cem\u003eTrichoderma spp.\u003c/em\u003e and synthetic chemical intervention were found non-significant (F\u003csub\u003e5,12\u003c/sub\u003e= 1.823, P\u0026thinsp;=\u0026thinsp;0.182) in relation to no intervention for total chlorophyll (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003ea) and carotenoids content (F\u003csub\u003e5,12\u003c/sub\u003e= 1.675, P\u0026thinsp;=\u0026thinsp;0.214; Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e3.3 Effects of \u003cem\u003eTrichoderma\u003c/em\u003e and insecticides on average fruit length/plant and yield/plant\u003c/h2\u003e \u003cp\u003eNo main effects were found significant between \u003cem\u003eTrichoderma\u003c/em\u003e or chemical interventions and no interventions on average fruit length (F\u003csub\u003e5,24\u003c/sub\u003e = 2.203, P\u0026thinsp;=\u0026thinsp;0.08; Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ea). The overall yield of cucumber was significantly increased with \u003cem\u003eTrichoderma\u003c/em\u003e and insecticides treatments than control (F\u003csub\u003e5,24\u003c/sub\u003e = 2.203, P\u0026thinsp;=\u0026thinsp;0.08; F\u003csub\u003e5,24\u003c/sub\u003e = 2.514, P\u0026thinsp;\u0026lt;\u0026thinsp;0.05; Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eb). Though no significant differences between \u003cem\u003eTrichoderma\u003c/em\u003e and insecticides were found but the highest fruit length was observed in \u003cem\u003eT. harzianum\u003c/em\u003e \u0026amp; CI\u0026thinsp;+\u0026thinsp;SI treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003ea) and highest yield was observed in \u003cem\u003eT. asperellum\u003c/em\u003e \u0026amp; CI treatment, respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003eb).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eOur findings in this study demonstrate that both \u003cem\u003eTrichoderma spp.\u003c/em\u003e are effective enough to control insect pests of cucumber while offering additional ecological benefits in contrast to conventional insecticides that documented in recent studies (Kubiak et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Monte, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Poveda, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The dual application of both \u003cem\u003eTrichoderma species\u003c/em\u003e not only suppressed tested insects; aphids, whiteflies, and red pumpkin beetle, but also enhanced trichome density, indicating a potential synergistic effect on plant defence mechanisms, and ultimately maintained a minimum number of diseased infested leaves (Kim, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Cucumber production around the world suffers considerable losses due to common sap-feeding pests such as aphids (\u003cem\u003eAphis gossypii\u003c/em\u003e), whiteflies (\u003cem\u003eBemisia tabaci\u003c/em\u003e), and red pumpkin beetle (\u003cem\u003eAulacophora foveicollis\u003c/em\u003e Lucas). However, the potential and sustainable management approach for these insect pests remain unexplored especially the role of \u003cem\u003eTrichoderma spp\u003c/em\u003e. in compare with conventional method. In this study, we documented that in parallel with conventional pesticides, the foliar application of \u003cem\u003eTrichoderma\u003c/em\u003e can also potentially minimize the attacks of aphids, whiteflies and red pumpkin beetle in cucumber.\u003c/p\u003e \u003cp\u003eAphid infestations are particularly damaging, as both nymphs and adults suck into the phloem, leading to leaf curling, discoloration, and in severe situations, wilting of entire plants, which translates into reduced fruit yield (Kumari et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Their sugary secretions, known as honeydew, foster the development of sooty mold, further interfering with photosynthesis and overall plant vitality (Razmjou et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Kumari et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Beyond direct injury, aphids also spread destructive viruses including Cucumber mosaic virus (CMV) in cucumber compounding yield losses through disease outbreaks. Our study demonstrated that the reduction of aphid number significantly reduced CMV diseases than insecticides and no intervention treatments suggested that aphids are highly resistant against insecticides due to their ability to produce genetically modified offspring that combat insecticides (Kaleem Ullah et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Edwards et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). In this study, both \u003cem\u003eTrichoderma spp.\u003c/em\u003e was successful to control aphids after foliar application assuming their direct effects on aphids through penetrating soft aphid body by puncturing cuticle and parasitize the host (Coppola et al. 2008; Battaglia et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Poveda, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Our study suggested that \u003cem\u003eTrichoderma spp.\u003c/em\u003e is a strong candidate to control cucumber aphids and CMV diseases that transmitted by aphids which is not successfully control by conventional approaches. In addition, \u003cem\u003eTrichoderma spp.\u003c/em\u003e may attract the natural enemies such parasitic wasp through releasing volatiles compounds that could also the reduced aphid population in \u003cem\u003eTrichoderma\u003c/em\u003e treated plants (Woo et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Poveda, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The ladybird beetle is the key predator to control aphids and has been found very effective in many studies (Omkar \u0026amp; Srivastava, 2023; Riddick, \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Ceryngier et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). If the ladybird beetle can be attracted by volatiles that released by \u003cem\u003eTrichoderma\u003c/em\u003e there likely an indirect plant defence to control aphids and CMV diseases, respectively. On the other hands, the conventional approach killed the target organism including non-target organisms such as ladybird beetle due to their wider mode of action (Bommarco et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Bielza, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). A future study hence require undertaking to explore the role of \u003cem\u003eTrichoderma spp.\u003c/em\u003e to attract natural enemies especially parasitic wasp or ladybird beetles to control aphids as indirect plant defence in field condition (Poveda, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eWhiteflies are also key pests in cucumber including many vegetables and fruit crops. They damage cucumber plants through depleting sap, which stunts growth and reduces photosynthetic activity, while also serving as carriers of viral diseases such as leaf curl virus in Tomato (Devendran et al. 2023). In this study, we found that the number of whiteflies significantly reduced with \u003cem\u003eT. harzianum\u003c/em\u003e that is slightly higher in insecticidal treatments. Interestingly, the effects of \u003cem\u003eT. asperellum\u003c/em\u003e on lowering whiteflies population remain unaffected that suggested species specific effects of \u003cem\u003eTrichoderma\u003c/em\u003e on hemipteran insect pest (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Our results indicated that we should be careful about the species-specific effects of \u003cem\u003eTrichoderma spp\u003c/em\u003e. before its application in field condition. However, the multiple application of \u003cem\u003eTrichoderma spp.\u003c/em\u003e could be able to minimize the limitation of species specificity of \u003cem\u003eTrichoderma\u003c/em\u003e through controlling wide range of insect pests and is associated plant diseases. While dual application of \u003cem\u003eTrichoderma spp.\u003c/em\u003e seems potential but it is important to keep in mind about the synergistic and antagonistic effects of the species within (Poveda, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eRed pumpkin beetles are also key pests for cucurbits production especially their gregarious feeding in young cucurbits seedlings (Hassan et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Cucumber and sweet gourds are the most suitable host for red pumpkin beetles at young and mature stage of plants, but they could feed on wide range of cucurbits host (Hassan et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Saljoqi et al. 2007). The beetles could defoliate the young leaves through chewing within a day and retard the growth of plants. The high infestation of red pumpkin beetles could cause yield loss up to 100% especially at seedling stage if there is no intervention take place (Regmi et al. 2020). The conventional contact insecticides are widely used to control the beetle, but it seems they became insecticides resistant over time (Hassan et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The alternative approach \u003cem\u003eTrichoderma spp.\u003c/em\u003e could be an option to minimize the pest damage but never been tested. Our study however, found that the population dynamics of beetles were significantly reduced with \u003cem\u003eT. harzianum\u003c/em\u003e suggested that the foliar application could increase cucumber trichomes density, antifeedant and repellent compounds, that could reduce the feeding of beetle population than control treatments in cucumber (Poveda, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The reduction of beetle population was consistent for both \u003cem\u003eTrichoderma spp.\u003c/em\u003e thereby, indicated that \u003cem\u003eTrichoderma\u003c/em\u003e likely to be an alternative option to manage red pumpkin beetle infestation, however field validation with large cucumber cultivar needed for validate the outcomes.\u003c/p\u003e \u003cp\u003eTrichomes are hair-like outgrowths that occur on nearly all non-aquatic plant parts, including leaves, stems, and even fruits (Goffreda et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1988\u003c/span\u003e). They may be glandular or non-glandular, unicellular or multicellular, and often function as a protective barrier against insects and fungal pathogens (Sarria et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Our study found that dual application of \u003cem\u003eTrichoderma spp.\u003c/em\u003e increases the number of trichome suggested to enhance plant protection and self-defense mechanism against sap feeding and chewing herbivores (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In addition to direct effects of \u003cem\u003eTrichoderma spp.\u003c/em\u003e on sap feeding and chewing herbivore, the increased number of trichome than control treatments suggested an additional benefits for plants that helps to combat insects and pathogens in cucumber. The number of trichomes also increased in insecticidal treatments than control however, our study not able provide explanation about the increase of trichome number in those treatments (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Our assumption is that plant respond to increase trichome number when additional crop protection strategy applied to control insect and pathogen attack. The increase of trichome number could negatively influence the feeding of sap feeding insects and chewing beetles, thereby the population of sap feeding and chewing insects reduced in this study with \u003cem\u003eTrichoderma\u003c/em\u003e. Previous study documents that trichomes can release sticky or viscous secretions that trap sap feeding insects or deter chewing herbivory by making feeding more difficult (Wagner, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Wheeler and Krimmel, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Some study also found that insects caught in these secretions may attract predators of herbivores, thereby strengthening the plant\u0026rsquo;s indirect defense system (Krimmel and Pearse, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Non-glandular trichomes, on the other hand, are often rigid or hooked structures that can physically puncture insect bodies or obstruct their movement, directly interfering with feeding behavior (Levin, 1973; Riddick and Simmons, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Hooked non-glandular types are particularly effective, as they can ensnare not only a wide range of herbivores but also their natural enemies (Riddick and Simmons, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Such entrapments can reduce the survival and reproductive success of insects and may even influence the long-term population dynamics and evolutionary fitness of the species involved (Peterson et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Riddick and Simmons, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eOur results also found that chlorophyll (primary photosynthetic pigment); and carotenoid content remain unaffected with treatment combination although we assume that there should be an increase in green tissues with \u003cem\u003eTrichoderma\u003c/em\u003e due to its positive effects on leaf parameters that documented in recent study (Paraginski et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). However, there are significant increase of average fruit length and yield of cucumber than control treatments that suggested the positive effects of \u003cem\u003eTrichoderma spp.\u003c/em\u003e on the production of cucumber. Although, there are similar results also found in insecticidal treatments, but cucumber produced in biopesticide treatments likely to prefer by consumer than insecticidal treatments due its less or no chemical residues. In nutshell, our research suggested the application of \u003cem\u003eTrichoderma spp\u003c/em\u003e. could an alternative option to fight insects and diseases than insecticides to grow sustainable and safe cucurbits for the consumer.\u003c/p\u003e"},{"header":"5. Conclusion","content":"\u003cp\u003eOur results concluded that both \u003cem\u003eT. asperellum\u003c/em\u003e and \u003cem\u003eT. harzianum\u003c/em\u003e were reduce the sap feeding and chewing herbivore population in cucumber than insecticidal and control treatments. \u003cem\u003eT. harzianum\u003c/em\u003e are likely to be more successful to control sap feeding and chewing herbivore including no negative effects on parasitoids population. The dual application of \u003cem\u003eTrichoderma spp.\u003c/em\u003e provided significant outcome through reducing the incidence of CMV diseases that suggested synergistic effects of \u003cem\u003eTrichoderma spp.\u003c/em\u003e The effects of insecticides treatment also found significant than control treatment, however, both treatments showed negative effects of parasitoids that suggested its effects on non-target organisms impede natural parasitism of sap feeding insects such as aphids. The increased of trichome number suggested additional benefits of \u003cem\u003eTrichoderma spp.\u003c/em\u003e to control pest density by reducing their feeding or repelling them from the leaves, however, more study need to conduct to explore this mechanism. Although our study did not find any positive effects of \u003cem\u003eTrichoderma spp.\u003c/em\u003e on photosynthetic tissues but future study focusing on the effects of \u003cem\u003eTrichoderma\u003c/em\u003e on leaf green tissues require to tease apart the mechanism. Although the yield parameters of cucumber positively affected by \u003cem\u003eTrichoderma\u003c/em\u003e and insecticidal treatments than control, but the negative effects of pesticides could be possible to minimize though using biopesticides as alternative option to control insects and disease of cucumber. Our study has boarder application on the management of cucurbits insects and diseases though considering biopesticides such as \u003cem\u003eTrichoderma\u003c/em\u003e as potential option while reducing the application of pesticides for safe and sustainable vegetables production for consumers.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eKH\u0026mdash; conceived, conceptualized, writing\u0026mdash;original draft preparation, writing\u0026mdash;review and editing, conduct experiments, data curation, analyzed the data, and prepared the figures. MMA- writing- original draft, review and editing. TS\u0026mdash;writing\u0026mdash;review and editing, and data curation. MFM\u0026mdash;writing\u0026mdash;review and editing, approved the final draft. SS- writing\u0026mdash;review and editing, approved the final draft. MP\u0026mdash;writing\u0026mdash;review and editing, approved the final draft.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAll data will be available upon reasonable request to corresponding author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research has been funded by UGC-SAURES authority from Sylhet Agricultural University. The gran number is SAURES AG-09.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor declarations\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in the paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe authors declare no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank Sylhet Agricultural Research Systems (SAURES; grant number SAURES-09) and University Grant Commission (UGC) for funding in this study. \u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAlın\u0026ccedil;, T. Cusumano, A. Peri, E. Torta, L. \u0026amp; Colazza, S. 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Lorito, M. \u003cem\u003eet al.\u003c/em\u003e \u003cem\u003eTrichoderma\u003c/em\u003e: a multipurpose, plant-beneficial microorganism for eco-sustainable agriculture. \u003cem\u003eNat Rev Microbiol\u003c/em\u003e \u003cstrong\u003e21\u003c/strong\u003e, 312\u0026ndash;326 (2023). https://doi.org/10.1038/s41579-022-00819-5\u003c/li\u003e\n \u003cli\u003eZaki, O. Weekers, F. Thonart, P. Tesch, E. Kuenemann, P. \u0026amp; Jacques, P. (2020). Limiting factors of mycopesticide development. \u003cem\u003eBiological Control\u003c/em\u003e, \u003cem\u003e144\u003c/em\u003e, 104220. https://doi.org/10.1016/j.biocontrol.2020.104220\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"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":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Insect community, Biopesticide, Cucumber, Insecticides, Trichoderma asperellum, Trichoderma harzianum","lastPublishedDoi":"10.21203/rs.3.rs-8410264/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8410264/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCucumbers (\u003cem\u003eCucumis sativa L\u003c/em\u003e) are economically important vegetable crops globally. However, it is highly susceptible to aphids, whiteflies and red pumpkin beetle that cause significant damage. Currently, chemical pesticide is practiced controlling herbivores, but it has severe health hazards. \u003cem\u003eTrichoderma \u003c/em\u003econsider as an eco-friendly management approach for controlling these herbivores. Hence, our study aims to compare the effectiveness of two \u003cem\u003eTrichoderma spp.\u003c/em\u003e and insecticides in controlling aphids, whiteflies and red pumpkin beetle of cucumber. We performed semi-field pot experiment with a cucumber variety (F1 hybrid cucumber) using two Trichoderma species (BP1-\u003cem\u003eTrichoderma asperellum\u003c/em\u003e (0R125623) @ 2 ´ 10\u003csup\u003e6\u003c/sup\u003e CFU/gm) and BP2- \u003cem\u003eTrichoderma harzianum\u003c/em\u003e 2 ´ 10\u003csup\u003e6\u003c/sup\u003e CFU/gm) @ 5gm/liter water), two insecticides (Cord 10 % EC and Cartap 50SP) including a control. Our results found that total insect community were higher in chemical intervention than \u003cem\u003eTrichoderma\u003c/em\u003e, showing a significant reduction of herbivorous insect. \u003cem\u003eTrichoderma harzianum\u003c/em\u003e showed higher efficacy for controlling herbivores pest density and percent infestation. The foliar application of \u003cem\u003eTrichoderma\u003c/em\u003e has positive effects on parasitoids. The incidence of cucumber mosaic virus (%) reduced to 8-10 % than control in dual application of \u003cem\u003eTrichoderma spp. \u003c/em\u003esuggested synergistic effects between species. Although total chlorophyll and carotenoids content in leaves remain unaffected, however, the average trichomes per leaves, fruit length and yield/plant significantly increased with both \u003cem\u003eTrichoderma \u003c/em\u003eand insecticidal treatments where \u003cem\u003eTrichoderma spp. \u003c/em\u003efound effective than insecticide treatment. Our research has broader implications for the management of cucurbits insects and diseases with less dependency on insecticides while considering \u003cem\u003eTrichoderma\u003c/em\u003e as a potential alternative.\u003c/p\u003e","manuscriptTitle":"The comparative assessment of Trichoderma and insecticides on chewing and phloem-feeding herbivores and physiological traits in cucumber","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-29 18:01:23","doi":"10.21203/rs.3.rs-8410264/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5acabe9a-9b49-43d8-8e3c-b19050de7e2a","owner":[],"postedDate":"January 29th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-04-29T13:24:41+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-29 18:01:23","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8410264","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8410264","identity":"rs-8410264","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}