Effect of thermotherapy on meristem culture response of two local cassava (Manihot esculenta Crantz) cultivars from Burkina Faso

Effect of thermotherapy on meristem culture response of two local cassava (Manihot esculenta Crantz) cultivars from Burkina Faso | 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 Effect of thermotherapy on meristem culture response of two local cassava ( Manihot esculenta Crantz) cultivars from Burkina Faso Wendpang-yidé Idrissa Caleb OUEDRAOGO, Koussao SOME, Rasmata NANA, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4969011/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 Using healthy cuttings can enhance the productivity of cultivars susceptible to viral diseases. However, due to the unavailability of healthy cuttings, farmers often have to rely on infected ones. This study aims to enhance the health quality of planting material by using thermotherapy and meristem culture techniques on two cassava cultivars. To achieve this, cuttings from the BFM206 and BFM260 cultivars were subjected to heat treatment in a thermotherapy chamber and a control treatment in field conditions. The heat treatment involved exposing the cuttings to a temperature of 37°C for the first week. The temperature was then increased by 2°C each week over four weeks, reaching 37°C, 39°C, 41°C, and 43°C, respectively. After the treatment, the resulting shoots were used as mother plants for meristem culture. The experimental design was a split-plot with 5 repetitions. The parameters studied included the severity and incidence of cassava mosaic disease, the regeneration rate, the callus formation rate, and growth parameters. The heat treatment resulted in a significant difference between the cultivars in terms of the severity of cassava mosaic disease (P < 0.001) and the regeneration rate (P = 0.008). Furthermore, compared to the control treatment, the heat treatment significantly increased the number of nodes by 4.92 and the number of stems by 1.42 in the BFM260 cultivar. In conclusion, thermotherapy can help reduce the severity of cassava mosaic disease in mother plants and thereby improve the in vitro growth of meristems. Agronomy Biotechnology and Bioengineering Food Science & Technology Manihot esculenta thermotherapy meristem regeneration growth Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction Burkina Faso is an agricultural country where 80% of the population earns their income from farming (MAAH, 2020 ). With a population of 20,487,979 in 2019 (INSD, 2019 ), more than 10% of whom rely on food assistance due to terrorism, Burkina Faso is grappling with food insecurity (Ouedraogo, 2023 ). In light of these difficulties, it is essential to diversify agriculture in Burkina Faso by integrating high-yield crops such as cassava. Cassava ( Manihot esculenta Crantz) is a perennial shrub used as an annual crop (Campos & Caligari, 2017 ). It is mainly grown for its edible starchy tuberous roots (Montagnac et al., 2009 ) and provides food for nearly one billion people worldwide (Otekunrin & Sawicka, 2019). Cassava is regarded as the most important crop after wheat, rice, maize, potatoes, and barley globally and plays a crucial role in food security in Africa (Saranraj et al., 2019 ). Africa is the leading producer of cassava, with over 193 million tons produced in 2020, representing nearly 64% of global output (FAO, 2022 ). In Burkina Faso, cassava cultivation is practiced across all regions (Soro, et al., 2021 ). Despite the growth in cassava cultivation, national production meets only about 18% of the demand (Plan Manioc, 2019 ). In fact, cassava production in Burkina Faso was over 10,000 tons in 2010 and reached 23,000 tons between 2013 and 2017 before declining to 17,000 tons in 2018 (FAO, 2022 ). This decline is attributed to several factors, including droughts and soil degradation in Burkina Faso (El-Sharkawy, 2004 ; Plan Manioc, 2019 ). Additionally, although vegetative propagation facilitates the spread of cassava, it also makes the crop susceptible to viruses, which pose a major threat to global production (Legg et al., 2014 ). The gradual accumulation of viruses in planting material over successive generations (Krishna et al., 2022 ) leads to decreased cassava yields. Among viral diseases, cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) are the most threatening, significantly impacting cassava production (Legg & Winter, 2020 , Shirima et al., 2019 ). In Burkina Faso, CMD is the most prevalent across all cassava growing regions (Soro et al., 2021 ; Tiendrébéogo et al., 2009 ). Moreover, the sweet cassava varieties cultivated in Burkina Faso are highly susceptible to CMD, resulting in low yields (Dabiré & Belem, 2003 ). These sweet varieties are vital for food security, as their tuber harvesting periods align with the lean season. Enhancing the productivity of these cultivars would therefore strengthen the resilience of small-scale farmers. However, improving the productivity of these cultivars requires bettering the sanitary quality of planting material (Chikoti et al., 2019 ). In this context, meristem culture and thermotherapy are effective approaches (Benke et al., 2023 ; Krishna et al., 2022 ; Nakabonge et al., 2020 ; Nehra & Kartha, 1994 o et al., 2020 ). This study aims to improve the quality of planting material through thermotherapy and meristem culture of two cassava cultivars. 2. Plant materiel The plant material used consisted of cuttings from two local cassava cultivars: BFM206 from Léo in the Centre-Ouest region, and BFM260 from Santidougou in the Haut-Bassins region. These cultivars come from the two major cassava-producing regions in Burkina Faso. They are characterized by a short growth cycle (6 to 8 months) and are highly favored by farmers for the ease of cooking their tuberous roots. Nonetheless, these cultivars are significantly affected by CMD. 3. Experimental setup The experimental setup used was a split-plot design. The main factor was the treatment, consisting of two levels: heat treatment (thermo-treated) and control treatment (untreated). The secondary factor was the cultivar, with two levels: BFM206 and BFM260. In total, 100 cuttings, 50 from each cultivar, were subjected to the heat treatment. Another 40 untreated cuttings, 20 from each cultivar, were planted under field conditions. In the laboratory, the meristems were cultured using the same experimental setup previously described. From the shoots of the heat-treated cuttings, 100 meristems, 50 per cultivar, were collected and cultured. For the control treatment, 40 meristems were collected and cultured. 4. Methods 4.1. Treatment of cuttings The heat treatment was carried out in a thermotherapy chamber. Initially, cuttings from the two cultivars were taken from plants exhibiting symptoms of cassava mosaic disease on their leaves. These cuttings were then planted in 750 ml pots filled with a substrate made from one volume of commercial potting soil and two volumes of sand, which had been sterilized at 120°C for 20 minutes. The pots were placed in the thermotherapy chamber for a 4-month incubation period. During the first week, the chamber temperature was set to 37°C, and it was subsequently increased to 39°C, 41°C, and 43°C in the second, third, and fourth weeks, respectively, with a weekly increment of 2°C, as per the method used by Hu et al. ( 2021 ). The chamber’s light intensity was 2000 lux with a photoperiod of 16 hours of light and 8 hours of darkness, following the procedure described by Hu et al. ( 2021 ). The relative humidity in the chamber ranged from 70–90%. For the control treatment, 20 cuttings per cultivar were planted in the field in 5 rows, with 4 cuttings per row. 4.2. Meristem culture The new shoots that emerged from the cuttings under heat treatment and under field conditions were used as mother plants for meristem culture. Meristem culture was conducted under sterile conditions. Explants (3 to 5 cm) taken from the apical regions of shoots developed from heat-treated and untreated cuttings were disinfected according to the method described by Ferguson et al. ( 2020 ). The explants were first immersed in 70% ethanol for 1 minute, followed by disinfection in a sodium hypochlorite solution containing 3.75 g L − 1 of active chlorine and two drops of Tween 20 for 15 minutes. The explants were then rinsed four times with sterile distilled water. After disinfecting the apical explants, meristems with 1 to 2 leaf primordia were carefully excised and cultured on a medium composed of elements recommended by Ferguson et al. ( 2020 ) and Graciela et al. ( 2010 ). The medium consisted of 4.43 g L − 1 of Murashige & Skoog ( 1962 ) basal medium (MS), supplemented with 30 g L − 1 of sucrose, 0.04 g L − 1 of 1-naphthaleneacetic acid, 0.08 g L − 1 of 6-benzylaminopurine, and 3 g L − 1 of phytagel. The pH of the medium was adjusted to 5.8 using 1N NaOH or HCl. The medium was sterilized at 120°C for 20 minutes. The meristems were then cultured in test tubes and incubated in a growth room for 4 weeks. The growth room provided light at 1500 lux with a photoperiod of 16 hours of light and 8 hours of darkness. The temperature was maintained at 27°C with a relative humidity of 65%. After 4 weeks, well-developed regenerated meristems were removed and transferred to new test tubes containing the same medium and incubated in the same growth room for an additional 6 weeks. 4.3. Measured parameters Parameters related to cassava mosaic disease included incidence (Inc) and severity (Sev), which were assessed at 2, 3, and 4 weeks after planting. Incidence was determined by counting the number of symptomatic plants. The percentage of symptomatic plants was calculated using the following Eq. (1): Cassava mosaic disease severity was assessed based on the observed symptoms on the plants using the scale from Soro et al. ( 2021 ), as follows: Score 1: plants with no symptoms Score 2: plants with mild symptoms Score 3: plants with moderate symptoms Score 4: plants with severe symptoms Score 5: plants with very severe symptoms Four (4) weeks after initiating meristem culture, parameters related to meristem response, including the regeneration rate of meristems (RRM) and callus formation rate (CFR), were calculated using equations (2) and (3), respectively: Growth parameters of plantlets generated from the meristems included the number of leaves (NL), the number of stems (NS), the number of nodes (NN), and the number of roots (NR). These parameters were measured by counting 6 weeks after transferring the regenerated meristems. 4.4. Statistical analysis Data entry and processing, including graph creation, were performed using Excel 2019. Two-factor analysis of variance (ANOVA) to explore interactions between cultivar and heat treatment, along with Tukey's HSD test at a 5% significance level, were conducted using RStudio software. 5. Results 5.1. Incidence and severity of cassava mosaic disease following heat treatment Under heat treatment, the incidence (Inc) and severity (Sev) of cassava mosaic disease in cultivar BFM206 displayed a sawtooth pattern over time (Fig. 1 A&B). The results showed that, initially at 100 ± 0% at 2 weeks after planting (WAP), the Inc decreased to 87.50 ± 4.33% at 3 WAP, then increased to 91.67 ± 4.87% at 4 WAP (Fig. 1 A). For Sev in cultivar BFM206, it was 2.39 ± 0.20 at 2 WAP, decreased to 1.95 ± 0.09 at 3 WAP, and increased to 2 ± 0.12 at 4 WAP (Fig. 1 B). In contrast, the Inc and Sev for cultivar BFM260 both decreased over time (Fig. 1 A&B). Indeed, the Inc for BFM260 fell from 68 ± 10.95% at 2 WAP to 60 ± 12.24% at 3 WAP, and further to 52 ± 8.36% at 4 WAP (Fig. 1 A). The Sev for BFM260 was 1.78 ± 0.10 at 2 WAP, 1.60 ± 0.12 at 3 WAP, and 1.52 ± 0.08 at 4 WAP (Fig. 1 B). The decrease in Inc over time between 2 and 4 WAP was not significant for cultivar BFM206 (P = 0.419) and for cultivar BFM260 (P = 0.164). For Sev, the reduction between 2 and 4 WAP was not significant for cultivar BFM206 (P = 0.162), but the decrease in Sev for cultivar BFM260 from 2 to 4 WAP was significant (P < 0.001). 5.2. Effect of treatment on meristem regeneration rate and callus formation rate Under the heat treatment, the meristem regeneration rates (RRM) observed for the BFM260 and BFM206 cultivars were 60.90 ± 12.58% and 34.50 ± 10.93%, respectively. The difference in RRM between the cultivars under heat treatment was significant (P = 0.008) (Fig. 2 A). In contrast, the RRM were similar (P = 1) between the cultivars under the control treatment, with 75 ± 16% for the BFM206 cultivar and 75 ± 13.67% for the BFM260 cultivar (Fig. 2 A). Additionally, for the BFM206 cultivar, the RRM under heat treatment differed significantly (P = 0.008) from the RRM under the control treatment (Fig. 2 A). Regarding the callus formation rate (CFR), it was 44.40 ± 15.49% and 46.70 ± 7.07% for the BFM206 and BFM260 cultivars, respectively, under heat treatment (Fig. 2 B). Under the control treatment, the BFM206 cultivar had an CFR of 50 ± 14.43%, and the BFM260 cultivar showed an CFR of 65 ± 17.50% (Fig. 2 B). The results of the mean separation analysis indicated that the CFR did not vary between the cultivars under heat treatment (P = 0.890) or under the control treatment (P = 0.337) (Fig. 2 B). Two-factor analyse of variance revealed a non-significant interaction between cultivar and treatment for both the RRM (P = 0.141) and the CFR (P = 0.577) (Table I). 5.3. Effect of treatment on meristem growth parameters Under the heat treatment, the number of nodes (NN) observed was 3.33 ± 0.57 nodes per meristem for cultivar BFM206 and 5.92 ± 2.36 nodes per meristem for cultivar BFM260 (Fig. 3 A). The difference in the NN between cultivars under the heat treatment was not significant (P = 0.303). In the control treatment, cultivar BFM260 had 1 ± 2.24 nodes per meristem, and cultivar BFM206 had 1.25 ± 2.50 nodes per meristem (Fig. 3 A). The difference in the NN between cultivars in the control treatment was also not significant (P = 0.998). Additionally, for cultivar BFM260, the NN under the heat treatment was significantly higher (P = 0.002) compared to the control treatment (Fig. 3 A). For the number of stems (NS), cultivars BFM206 and BFM260 recorded 1.67 ± 0.57 and 2.62 ± 0.60 stems per meristem, respectively, under the heat treatment (Fig. 3 B). The mean separation indicated no significant difference (P = 0.467) in the NS between the cultivars under the heat treatment. In the control treatment, cultivar BFM206 had 1 ± 0.1 stems per meristem and cultivar BFM260 had 1.20 ± 0.44 stems per meristem (Fig. 3 B). The difference in NS between cultivars in the control treatment was also not significant (P = 0.990). The results showed that for cultivar BFM260, the NS under the heat treatment was significantly different (P = 0.05) from the control treatment. Regarding the number of roots (NR), cultivars BFM260 and BFM206 developed 1.31 ± 2.14 and 0.33 ± 0.57 roots per meristem, respectively, under the heat treatment (Fig. 3 C). The mean separation showed no significant difference (P = 0.786) in the NR between cultivars under the heat treatment. In the control treatment, no root development was observed for either cultivar (Fig. 3 C). For the number of leaves (NL), cultivars BFM206 and BFM260 produced 2.67 ± 0.57 and 2.85 ± 1.14 leaves per meristem, respectively, under the heat treatment. The difference in the NL between cultivars under the heat treatment was not significant (P = 0.994). In the control treatment, cultivar BFM206 had 3.75 ± 0.95 leaves per meristem, and cultivar BFM260 had 2.40 ± 1.34 leaves per meristem (Fig. 3 D). The difference in the NL between cultivars in the control treatment was also not significant (P = 0.302). The two-factor analyse of variance revealed a non-significant cultivar*treatment interaction for the NN (P = 0.188), the NS (P = 0.424), the NR (P = 0.525), and the NL (P = 0.156) (Table I). Table I. Means and two-factor analysis of variance of measured parameters. Factors/Levels MRR (%) CFR (%) NN NL NS NR Cultivars (C) BFM206 45.33 ± 15.20 46.80 ± 33 2.14 ± 2.12 3.29 ± 0.95 1.29 ± 0.48 0.14 ± 0.37 BFM260 65.20 ± 12.11 57.10 ± 23.07 4.56 ± 3.20 2.72 ± 1.18 2.22 ± 1.26 0.94 ± 1.89 P (α = 0.05) 0.018 0.354 0.039 0.272 0.041 0.525 Treatments (T) Untreated 75 ± 19.76 57.50 ± 24.36 1.11 ± 2.20 3 ± 1.32 1.11 ± 0.33 0 Thermo-treated 46.50 ± 3.24 45.20 ± 21.94 5.44 ± 2.37 2.81 ± 1.50 2.44 ± 1.21 1.13 ± 1.96 P (α = 0.05) 0.002 0.267 < 0.001 0.546 0.004 0.289 C*T 0.141 0.577 0.188 0.156 0.424 0.525 MRR: meristem regeneration rate; CFR: callus formation rate; NN: number of nodes; NL: number of leaves; NS: number of stems; NR: number of roots; P: probability; α: significance level; C*T: cultivar*treatment interaction; mean ± standard deviation 6. Discussion The heat treatment was applied to cuttings infected with cassava mosaic virus. The observed reduction in both incidence and severity of cassava mosaic disease over time on the cuttings after thermal treatment can be explained by the heat-induced reduction in viral activity. Kouassi et al., ( 2019 ) and Zinga et al. ( 2014 ) have shown that thermotherapy significantly decreases both the incidence and severity of cassava mosaic disease. Heat limits the replication and movement of viruses within the host (Panattoni et al., 2013 ; Wang et al., 2018 ) leading to a reduction in symptom expression. The meristems obtained from shoots both thermo-treated and untreated cuttings were cultured. The significant difference in meristem regeneration rates between the cultivars under heat treatment suggests that the BFM260 meristems exhibit better heat tolerance compared to the BFM206 meristems. Moreover, thermotherapy had a detrimental effect on the regeneration ability of BFM206 meristems compared to the control treatment. These results are consistent with Wang et al. ( 2018 ), who found that high and prolonged thermal treatments can diminish the regeneration capacity of apices taken from treated shoots. Additionally, Magyar-Tábori et al. ( 2021 ) proposed that this reduction in regeneration rate under thermal stress could be due to oxidative damage, which affects cellular metabolism and may lead to cell death. However, the lack of a significant interaction between cultivar and treatment on meristem regeneration indicates that thermotherapy impacts the regeneration of meristems similarly across both cultivars. Therefore, the observed difference in regeneration rates under thermal treatment is more likely related to the inherent regenerative capacity of the meristems of each cultivar rather than their ability to tolerate the thermal treatment. Meristem regeneration was frequently associated with callus formation. The callus formation rates observed, ranging between 44.40% and 65%, were relatively high. This suggests that the genetic stability of the resulting plantlets needs to be evaluated. Indeed, high callus formation can lead to somaclonal variations, potentially resulting in genetically diverse plantlets (Amitchihoué et al., 2019 ; George et al., 2008 ) The regenerated meristems gave rise to plantlets. The superior performance in terms of the number of nodes and stems observed in the BFM260 cultivar may be attributed to the quality of the culture medium. The medium used appears to be better suited for the growth of BFM260 plantlets compared to those of the BFM206 cultivar. Indeed, different species or varieties have varying requirements for nutrients and growth hormones (Carvalho et al., 2017 ; Uwimana et al., 2022 ). Additionally, the higher number of nodes and stems in BFM260 cultivar under heat treatment may also be linked to its low severity of cassava mosaic disease. The reduction in disease severity likely resulted in virus-free meristems, which would have grown better than those from untreated plants. Indeed, during thermal treatment, the replication and movement of viruses to the host’s meristematic cells are inhibited by heat (Panattoni et al., 2013 ; Wang et al., 2018 ; Wang et al., 2008 ), thus promoting better in vitro growth. In contrast, untreated plants would produce highly infected meristems, which could lead to reduced growth of plantlets in the control treatment. The inhibition of in vitro growth due to viral infection has been documented in potato ( Solanum tuberosum L.) and apple ( Pyrus spp .) (Chen et al., 2017 ; Li et al. ( 2013 ). Furthermore, the absence of roots in plantlets from both cultivars under the control treatment may be due to the meristems' inability to utilize auxin present in the culture medium. This inability is likely due to high virus levels in meristems from untreated shoots. Auxin is essential for root formation (Cammarata et al., 2023 ; Kurepa & Smalle, 2022 ), and its ineffective use and absorption would impair root development (Chen et al., 2017 ). Wang et al. ( 2009 ) also noted that virus-induced difficulty in auxin absorption by plantlets affects normal cellular function, further impeding root development. 7. Conclusion This study evaluated the effects of heat treatment on cassava cuttings in relation to the severity and incidence of cassava mosaic disease, as well as its impact on meristem regeneration and the growth of plantlets derived from meristem cultures. Indeed, the findings revealed that heat treatment of cuttings significantly reduced meristem regeneration rates in the BFM206 cultivar. For the BFM260 cultivar, thermal treatment of cuttings effectively decreased the severity of cassava mosaic disease. Additionally, heat treatment of cuttings enhanced the number of nodes and stems in plantlets of the BFM260 cultivar. Future research should explore optimal culture media to further improve the multiplication and growth of these plantlets. Abbreviations BAP 6-Benzylamino purine CMD Cassava Mosaic Disease CBSD Cassava Brown Streak Disease MS Murashige and Skoog medium NAA 1-Naphthaleneacetic acid Declarations Conflicts of interests The authors declare that there are no conflicts of interests regarding the publication of this article. Acknowledgements The authors are grateful to the “Ministère de l’Enseignement Supérieur, de la Recherche et de l’Innovation (MESRI)” through the “Programme d’Appui à l’Enseignement Supérieur (PAES)” for funding this work. We also wish to thank the “Institute de l’Environnement et de la Recherche Agricole (INERA)” for providing the plant material and hosting this work. References Amitchihoué FS, Jane WK, Elijah A, Jerome AH, Colombe D, Peter N, Corneille A (2019) Callus induction in three mosaic disease resistant cassava cultivars in Benin and genetic stability of the induced calli using simple sequence repeat (SSR) and sequence-characterized amplified region (SCAR) markers. Afr J Biotechnol 18:1044–1053. 10.5897/ajb2019.16967 Benke AP, Krishna R, Khandagale K, Gawande S, Shelke P, Dukare S, Dhumal S, Singh M, Mahajan V (2023) Efficient elimination of viruses from garlic using a combination of shoot meristem culture, thermotherapy, and chemical treatment. Pathogens 12. 10.3390/pathogens12010129 Cammarata J, Roeder AHK, Scanlon MJ (2023) The ratio of auxin to cytokinin controls leaf development and meristem initiation in Physcomitrium patens . J Exp Bot 74:6541–6550. 10.1093/jxb/erad299 Campos H, Caligari PDS (2017) Chapter 5: Cassava ( Manihot esculenta Crantz) in genetic improvement of tropical crops. In Genetic Improvement of Tropical Crops. 1–52. 10.1007/978-3-319-59819-2 Carvalho MJS, Oliveira EJ, Souza AS, Pereira JS, Diamantino MSAS, Oliveira SAS (2017) Cleaning cassava genotypes infected with cassava frogskin disease via in vitro shoot tip culture. Genet Mol Res 16:1–17. 10.4238/gmr16029556 Chen J, Tang HH, Li L, Qin SJ, Wang GP, Hong N (2017) Effects of virus infection on plant growth, root development and phytohormone levels in in vitro-cultured pear plants. Planr Cell Tissue Organ Cult 131:359–368. 10.1007/s11240-017-1289-1 Chikoti PC, Mulenga RM, Tembo M, Sseruwagi P (2019) Cassava mosaic disease: a review of a threat to cassava production in Zambia. J Plant Pathol 101:479–479. 10.1007/s42161-019-00267-w Dabiré R, Belem J (2003) Les plantes à tubercules et racines au Burkina Faso. Wasnet 8:12–16 El-Sharkawy MA (2004) Cassava biology and physiology. Plant Mol Biol 56:481–501. 10.1007/s11103-005-2270-7 FAO (2022) Food and Agriculture Organization of the United Nations. FAOSTAT statistics database, Rome Ferguson M, Choti A, Munguti F (2020) Virus Elimination and Testing in Cassava: A pratical manual. IITA, Ibadan, Nigeria. : 1–24 George E, Hall M, De Klerk J (2008) Plant propagation by tissue culture 3rd edition Volume 1. The backgorund Graciela MB, Roa CJE, Aranzales ER, Debouck DG (2010) Handbook of procedures for in vitro germplasm conservation Hu G jun, Dong Y, feng, Zhang Z ping, Fan X, dong, Ren F (2021) Elimination of Grapevine fleck virus and Grapevine rupestris stem pitting-associated virus from Vitis vinifera 87 – 1 by ribavirin combined with thermotherapy. Journal of Integrative Agriculture 20: 2463–2470. 10.1016/S2095-3119(20)63336-6 INSD (2019) Cinquième recensement général de la population et de l’habitation du Burkina Faso. synthèse des résultats définitifs Kouassi MK, Kouassi N, Yeo FE (2019) Caractéristiques d’un dispositif de thermothérapie construit localement pour l’assainissement de plants de manioc. AISA Développement n° 16:2–5. 10.13140/RG.2.2.14034.53448 Krishna R, Ansari WA, Khandagale K, Benke AP, Soumia PS, Manjunathagowda DC, Gawande SJ, Ade AB, Mokat DN, Singh M (2022) Meristem culture: a potential technique for in vitro virus-free plants production in vegetatively propagated crops. Advances in Plant Tissue Culture. Elsevier, pp 325–343. DOI: 10.1016/B978-0-323-90795-8.00017-5 Kurepa J, Smalle JA (2022) Auxin/Cytokinin antagonistic control of the shoot/root growth ratio and its relevance for adaptation to drought and nutrient deficiency stresses. Int J Mol Sci 23. 10.3390/ijms23041933 Legg J, Winter S (2020) Cassava mosaic viruses ( Geminiviridae ). Encyclopedia of virology. Elsevier, pp 301–312. DOI: 10.1016/B978-0-12-809633-8.21523-9 Legg JP, Lava Kumar P, Makeshkumar T, Tripathi L, Ferguson M, Kanju E, Ntawuruhunga P, Cuellar W (2014) Cassava virus diseases. Advances in Virus Research. Elsevier Inc., pp 85–142. DOI: 10.1016/bs.aivir.2014.10.001 Li JW, Wang B, Song XM, Wang RR, Chen L, Zhang H, Zhang ZB, Wang QC (2013) Potato leafroll virus (PLRV) and Potato virus Y (PVY) influence vegetative growth, physiological metabolism, and microtuber production of in vitro-grown shoots of potato ( Solanum tuberosum L.). Plant Cell, Tissue and Organ Culture 114: 313–324. 10.1007/s11240-013-0327-x MAAH (2020) Rapport de l’enquête nationale intégré de sécurité alimentaire et nutritionnelle, Burkina Faso Magyar-Tábori K, Mendler-Drienyovszki N, Hanász A, Zsombik L, Dobránszki J (2021) Phytotoxicity and other adverse effects on the in vitro shoot cultures caused by virus elimination treatments: reasons and solutions. Plants 10:670. 10.3390/plants10040670 Plan manioc (2019) Plan d’actions pour le développement de la Filière. manioc du Burkina Faso Montagnac JA, Davis CR, Tanumihardjo SA (2009) Nutritional value of cassava for use as a staple food and recent advances for improvement. Compr Rev Food Sci Food Saf 8:181–194. 10.1111/j.1541-4337.2009.00077.x Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497 Nakabonge G, Nangonzi R, Tumwebaze BS, Kazibwe A, Samukoya C, Baguma Y (2020) Production of virus-free cassava through hot water therapy and two rounds of meristem tip culture. Cogent Food Agric 6:1800923. 10.1080/23311932.2020.1800923 Nehra NS, Kartha KK (1994) Meristem and shoot tip culture: requirements and applications. Plant cell and tissue culture. Dordrecht, Springer Netherlands, pp 37–70. DOI: 10.1007/978-94-017-2681-8_3 Olutosin AO, Sawicka B (2019) Cassava, a 21st century staple crop: how can Nigeria harness its enormous trade potentials? Acta Sci Agric 3:194–202. 10.31080/asag.2019.03.0586 Ouedraogo A (2023) Insécurité alimentaire et résilience au Burkina Faso: une analyse comparée des ménages des personnes déplacées internes (pdi) des centres d’accueil et des ménages locaux dans la commune Panattoni A, Luvisi A, Triolo E (2013) Review. Elimination of viruses in plants: twenty years of progress. Span J Agricultural Res 11:173–188. 10.5424/sjar/2013111-3201 Saranraj P, Behera SS, Ray RC (2019) Chapter 7 - Traditional foods from tropical root and tuber crops: innovations and challenges. Innovations in traditional foods, Galanakis CMBT-I in TF. Woodhead Publishing, pp 159–191. DOI: https://doi.org/10.1016/B978-0-12-814887-7.00007-1 Shirima RR, Maeda DG, Kanju EE, Tumwegamire S, Ceasar G, Mushi E, Sichalwe C, Mtunda K, Mkamilo G, Legg JP (2019) Assessing the degeneration of cassava under high-virus inoculum conditions in Coastal Tanzania. Plant Dis 103:2652–2664. 10.1094/PDIS-05-18-0750-RE Soro M, Tiendrébéogo F, Pita JS, Traoré ET, Somé K, Tibiri EB, Néya JB, Mutuku JM, Simporé J, Koné D (2021) Epidemiological assessment of cassava mosaic disease in Burkina Faso. Plant Pathol 70:2207–2216. 10.1111/ppa.13459 Su YH, Liu YB, Zhang XS (2011) Auxin-cytokinin interaction regulates meristem development. Mol Plant 4:616–625. 10.1093/mp/ssr007 Tiendrébéogo F, Lefeuvre P, Hoareau M, Traoré VSE, Barro N, Reynaud B, Traoré AS, Konaté G, Traoré O, Lett J-M (2009) Occurrence of East African cassava mosaic virus‐Uganda (EACMV‐UG) in Burkina Faso. Plant Pathol 58:783–783. 10.1111/j.1365-3059.2009.02110.x Uwimana J, Sakha M, Danga B, Gitari H, Gweyi-Onyango JP (2022) Involvement of plant growth regulators and varieties in the multiplication of cassava planting materials: a case study of Rwanda. Int J Bioresource Sci 9. 10.30954/2347-9655.02.2022.14 Wang CX, Hong N, Wang GP, Jiang B, Fan XD (2009) Effects of c itrus tristeza virus on the growth of in vitro-cultured citrus. J Plant Pathol 91:357–363 Wang MR, Cui ZH, Li JW, Hao XY, Zhao L, Wang QC (2018) vitro thermotherapy-based methods for plant virus eradication. Plant Methods 14:1–18. 10.1186/s13007-018-0355-y Wang Q, Cuellar WJ, Rajamäki ML, Hirata Y, Valkonen JPT (2008) Combined thermotherapy and cryotherapy for efficient virus eradication: relation of virus distribution, subcellular changes, cell survival and viral RNA degradation in shoot tips. Mol Plant Pathol 9:237–250. 10.1111/j.1364-3703.2007.00456.x Yéo EF, Kouassi MK, Pita JS, Kouassi NK, Koné D, N’guetta SPA (2020) Using thermotherapy and meristem tip culture for producing virus-free cassava planting material from six varieties cultivated in Côte d’ivoire. Int J Sci Technol Res 9:1607–1612 Zinga I et al (2014) Field evaluation of the effectiveness of thermotherapy against cassava mosaic disease in Central African Republic. Am J Experimental Agric 4:1232–1241. 10.9734/ajea/2014/10275 Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted 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-4969011","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":344542535,"identity":"796e1f00-e015-4e6a-b007-a72f7c08ed06","order_by":0,"name":"Wendpang-yidé Idrissa Caleb OUEDRAOGO","email":"data:image/png;base64,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","orcid":"https://orcid.org/0009-0006-4917-5321","institution":"Université Joseph Ki-Zerbo","correspondingAuthor":true,"prefix":"","firstName":"Wendpang-yidé","middleName":"Idrissa Caleb","lastName":"OUEDRAOGO","suffix":""},{"id":344543428,"identity":"f34a322e-e0ef-4630-becc-1efba4e3184c","order_by":1,"name":"Koussao SOME","email":"","orcid":"","institution":"Institut de l’Environnement et de Recherches Agricoles (INERA)","correspondingAuthor":false,"prefix":"","firstName":"Koussao","middleName":"","lastName":"SOME","suffix":""},{"id":344543429,"identity":"55ada2a9-badc-4332-bf1e-d8ca13fab7d2","order_by":2,"name":"Rasmata NANA","email":"","orcid":"","institution":"Université Joseph Ki-Zerbo","correspondingAuthor":false,"prefix":"","firstName":"Rasmata","middleName":"","lastName":"NANA","suffix":""},{"id":344543430,"identity":"5f0cf21d-6065-438f-8239-eadc3e376a05","order_by":3,"name":"Djakaridja TIAMA","email":"","orcid":"","institution":"Institut de l’Environnement et de Recherches Agricoles (INERA)","correspondingAuthor":false,"prefix":"","firstName":"Djakaridja","middleName":"","lastName":"TIAMA","suffix":""},{"id":344543431,"identity":"5e38d8b4-e257-438b-a8b1-92984a67da3c","order_by":4,"name":"Ousséni BERTHE","email":"","orcid":"","institution":"Université Joseph Ki-Zerbo","correspondingAuthor":false,"prefix":"","firstName":"Ousséni","middleName":"","lastName":"BERTHE","suffix":""},{"id":344543432,"identity":"47f35643-f437-4d82-be6b-b2c390c2dde2","order_by":5,"name":"Monique SORO","email":"","orcid":"","institution":"Institut de l’Environnement et de Recherches Agricoles (INERA)","correspondingAuthor":false,"prefix":"","firstName":"Monique","middleName":"","lastName":"SORO","suffix":""}],"badges":[],"createdAt":"2024-08-24 11:52:13","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-4969011/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4969011/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":63362134,"identity":"22c0ae06-d599-4984-8884-8b4bb93aeb5d","added_by":"auto","created_at":"2024-08-27 10:28:35","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":89045,"visible":true,"origin":"","legend":"\u003cp\u003eEvolution over time of means of the incidence (A) and severity (B) of cassava mosaic disease in two cultivars subjected to heat treatment. Means sharing the same letter are not significantly different at the 5 % level according to Tukey's test.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4969011/v1/5dd573030f38b5ab76c015f0.png"},{"id":63362133,"identity":"b51f9718-da13-4aee-96ec-173e6e63bd5b","added_by":"auto","created_at":"2024-08-27 10:28:35","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":110397,"visible":true,"origin":"","legend":"\u003cp\u003eMeans of meristem regeneration rates (A) and callus formation rates (B) of cultivars under different treatments. Means sharing the same letter are not significantly different at the 5 % level by Tukey's test.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4969011/v1/875e448d9d64e80201fc525f.png"},{"id":63362132,"identity":"9e36c589-abb7-4ec6-a168-8b7e4b8fdeb2","added_by":"auto","created_at":"2024-08-27 10:28:35","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":90621,"visible":true,"origin":"","legend":"\u003cp\u003eMeans of number of nodes (A), number of stems (B), number of roots (C), and number of leaves (D) of the cultivars under different treatments. Means with the same letter are not significantly different at the 5 % level according to Tukey's test.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4969011/v1/81287c6787f613894129b9fe.png"},{"id":63362135,"identity":"75d2d40e-b7a0-49db-a13a-5b4ed9b9eb73","added_by":"auto","created_at":"2024-08-27 10:28:36","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":523042,"visible":true,"origin":"","legend":"\u003cp\u003eMeans of regenerated meristems at 4 weeks after culturing (A); plantlets with roots, stems, and leaves at 6 weeks after transfer of the meristems (B).\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4969011/v1/605dd13725d1e756124bb3dc.png"},{"id":63362830,"identity":"a7aac5de-d3d5-400b-84d3-f1c937b6f629","added_by":"auto","created_at":"2024-08-27 10:36:35","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1451254,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4969011/v1/09b41203-3440-49e0-9c4d-fba94363a07e.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eEffect of thermotherapy on meristem culture response of two local cassava (\u003cem\u003eManihot esculenta\u003c/em\u003e Crantz) cultivars from Burkina Faso\u003c/p\u003e","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eBurkina Faso is an agricultural country where 80% of the population earns their income from farming (MAAH, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). With a population of 20,487,979 in 2019 (INSD, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), more than 10% of whom rely on food assistance due to terrorism, Burkina Faso is grappling with food insecurity (Ouedraogo, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). In light of these difficulties, it is essential to diversify agriculture in Burkina Faso by integrating high-yield crops such as cassava. Cassava (\u003cem\u003eManihot esculenta\u003c/em\u003e Crantz) is a perennial shrub used as an annual crop (Campos \u0026amp; Caligari, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). It is mainly grown for its edible starchy tuberous roots (Montagnac et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) and provides food for nearly one billion people worldwide (Otekunrin \u0026amp; Sawicka, 2019). Cassava is regarded as the most important crop after wheat, rice, maize, potatoes, and barley globally and plays a crucial role in food security in Africa (Saranraj et al., \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Africa is the leading producer of cassava, with over 193\u0026nbsp;million tons produced in 2020, representing nearly 64% of global output (FAO, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In Burkina Faso, cassava cultivation is practiced across all regions (Soro, et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Despite the growth in cassava cultivation, national production meets only about 18% of the demand (Plan Manioc, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In fact, cassava production in Burkina Faso was over 10,000 tons in 2010 and reached 23,000 tons between 2013 and 2017 before declining to 17,000 tons in 2018 (FAO, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). This decline is attributed to several factors, including droughts and soil degradation in Burkina Faso (El-Sharkawy, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Plan Manioc, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Additionally, although vegetative propagation facilitates the spread of cassava, it also makes the crop susceptible to viruses, which pose a major threat to global production (Legg et al., \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). The gradual accumulation of viruses in planting material over successive generations (Krishna et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) leads to decreased cassava yields. Among viral diseases, cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) are the most threatening, significantly impacting cassava production (Legg \u0026amp; Winter, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2020\u003c/span\u003e, Shirima et al., \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In Burkina Faso, CMD is the most prevalent across all cassava growing regions (Soro et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Tiendr\u0026eacute;b\u0026eacute;ogo et al., \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Moreover, the sweet cassava varieties cultivated in Burkina Faso are highly susceptible to CMD, resulting in low yields (Dabir\u0026eacute; \u0026amp; Belem, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). These sweet varieties are vital for food security, as their tuber harvesting periods align with the lean season. Enhancing the productivity of these cultivars would therefore strengthen the resilience of small-scale farmers. However, improving the productivity of these cultivars requires bettering the sanitary quality of planting material (Chikoti et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). In this context, meristem culture and thermotherapy are effective approaches (Benke et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Krishna et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Nakabonge et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Nehra \u0026amp; Kartha, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e1994\u003c/span\u003eo et al., \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). This study aims to improve the quality of planting material through thermotherapy and meristem culture of two cassava cultivars.\u003c/p\u003e"},{"header":"2. Plant materiel","content":"\u003cp\u003eThe plant material used consisted of cuttings from two local cassava cultivars: BFM206 from L\u0026eacute;o in the Centre-Ouest region, and BFM260 from Santidougou in the Haut-Bassins region. These cultivars come from the two major cassava-producing regions in Burkina Faso. They are characterized by a short growth cycle (6 to 8 months) and are highly favored by farmers for the ease of cooking their tuberous roots. Nonetheless, these cultivars are significantly affected by CMD.\u003c/p\u003e"},{"header":"3. Experimental setup","content":"\u003cp\u003eThe experimental setup used was a split-plot design. The main factor was the treatment, consisting of two levels: heat treatment (thermo-treated) and control treatment (untreated). The secondary factor was the cultivar, with two levels: BFM206 and BFM260. In total, 100 cuttings, 50 from each cultivar, were subjected to the heat treatment. Another 40 untreated cuttings, 20 from each cultivar, were planted under field conditions. In the laboratory, the meristems were cultured using the same experimental setup previously described. From the shoots of the heat-treated cuttings, 100 meristems, 50 per cultivar, were collected and cultured. For the control treatment, 40 meristems were collected and cultured.\u003c/p\u003e"},{"header":"4. Methods","content":"\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n \u003ch2\u003e4.1. Treatment of cuttings\u003c/h2\u003e\n \u003cp\u003eThe heat treatment was carried out in a thermotherapy chamber. Initially, cuttings from the two cultivars were taken from plants exhibiting symptoms of cassava mosaic disease on their leaves. These cuttings were then planted in 750 ml pots filled with a substrate made from one volume of commercial potting soil and two volumes of sand, which had been sterilized at 120°C for 20 minutes. The pots were placed in the thermotherapy chamber for a 4-month incubation period. During the first week, the chamber temperature was set to 37°C, and it was subsequently increased to 39°C, 41°C, and 43°C in the second, third, and fourth weeks, respectively, with a weekly increment of 2°C, as per the method used by Hu et al. (\u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e). The chamber’s light intensity was 2000 lux with a photoperiod of 16 hours of light and 8 hours of darkness, following the procedure described by Hu et al. (\u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e). The relative humidity in the chamber ranged from 70–90%. For the control treatment, 20 cuttings per cultivar were planted in the field in 5 rows, with 4 cuttings per row.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003e4.2. Meristem culture\u003c/h2\u003e\n \u003cp\u003eThe new shoots that emerged from the cuttings under heat treatment and under field conditions were used as mother plants for meristem culture. Meristem culture was conducted under sterile conditions. Explants (3 to 5 cm) taken from the apical regions of shoots developed from heat-treated and untreated cuttings were disinfected according to the method described by Ferguson et al. (\u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e). The explants were first immersed in 70% ethanol for 1 minute, followed by disinfection in a sodium hypochlorite solution containing 3.75 g L\u003csup\u003e− 1\u003c/sup\u003e of active chlorine and two drops of Tween 20 for 15 minutes. The explants were then rinsed four times with sterile distilled water. After disinfecting the apical explants, meristems with 1 to 2 leaf primordia were carefully excised and cultured on a medium composed of elements recommended by Ferguson et al. (\u003cspan class=\"CitationRef\"\u003e2020\u003c/span\u003e) and Graciela et al. (\u003cspan class=\"CitationRef\"\u003e2010\u003c/span\u003e). The medium consisted of 4.43 g L\u003csup\u003e− 1\u003c/sup\u003e of Murashige \u0026amp; Skoog (\u003cspan class=\"CitationRef\"\u003e1962\u003c/span\u003e) basal medium (MS), supplemented with 30 g L\u003csup\u003e− 1\u003c/sup\u003e of sucrose, 0.04 g L\u003csup\u003e− 1\u003c/sup\u003eof 1-naphthaleneacetic acid, 0.08 g L\u003csup\u003e− 1\u003c/sup\u003e of 6-benzylaminopurine, and 3 g L\u003csup\u003e− 1\u003c/sup\u003eof phytagel. The pH of the medium was adjusted to 5.8 using 1N NaOH or HCl. The medium was sterilized at 120°C for 20 minutes. The meristems were then cultured in test tubes and incubated in a growth room for 4 weeks. The growth room provided light at 1500 lux with a photoperiod of 16 hours of light and 8 hours of darkness. The temperature was maintained at 27°C with a relative humidity of 65%. After 4 weeks, well-developed regenerated meristems were removed and transferred to new test tubes containing the same medium and incubated in the same growth room for an additional 6 weeks.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n \u003ch2\u003e4.3. Measured parameters\u003c/h2\u003e\n \u003cp\u003eParameters related to cassava mosaic disease included incidence (Inc) and severity (Sev), which were assessed at 2, 3, and 4 weeks after planting. Incidence was determined by counting the number of symptomatic plants. The percentage of symptomatic plants was calculated using the following Eq.\u0026nbsp;(1):\u003c/p\u003e\n \u003cdiv id=\"Equa\" class=\"Equation\"\u003e\n \u003cdiv class=\"mathdisplay\" id=\"FileID_Equa\" name=\"EquationSource\"\u003e\u003cimg src=\"data:image/png;base64,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\" width=\"398\" height=\"40\"\u003e\u003c/div\u003e\n \u003c/div\u003e\n \u003cp\u003eCassava mosaic disease severity was assessed based on the observed symptoms on the plants using the scale from Soro et al. (\u003cspan class=\"CitationRef\"\u003e2021\u003c/span\u003e), as follows:\u003c/p\u003e\n \u003cul\u003e\n \u003cli\u003e\n \u003cp\u003eScore 1: plants with no symptoms\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eScore 2: plants with mild symptoms\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eScore 3: plants with moderate symptoms\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eScore 4: plants with severe symptoms\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003eScore 5: plants with very severe symptoms\u003c/p\u003e\n \u003c/li\u003e\n \u003c/ul\u003e\n \u003cp\u003eFour (4) weeks after initiating meristem culture, parameters related to meristem response, including the regeneration rate of meristems (RRM) and callus formation rate (CFR), were calculated using equations (2) and (3), respectively:\u003c/p\u003e\n \u003cp\u003e\u003cimg src=\"data:image/png;base64,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\"\u003e\u003cbr\u003e\u003c/p\u003e\n \u003cp\u003eGrowth parameters of plantlets generated from the meristems included the number of leaves (NL), the number of stems (NS), the number of nodes (NN), and the number of roots (NR). These parameters were measured by counting 6 weeks after transferring the regenerated meristems.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003e4.4. Statistical analysis\u003c/h2\u003e\n \u003cp\u003eData entry and processing, including graph creation, were performed using Excel 2019. Two-factor analysis of variance (ANOVA) to explore interactions between cultivar and heat treatment, along with Tukey's HSD test at a 5% significance level, were conducted using RStudio software.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"5. Results","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e5.1. Incidence and severity of cassava mosaic disease following heat treatment\u003c/h2\u003e \u003cp\u003eUnder heat treatment, the incidence (Inc) and severity (Sev) of cassava mosaic disease in cultivar BFM206 displayed a sawtooth pattern over time (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA\u0026amp;B). The results showed that, initially at 100\u0026thinsp;\u0026plusmn;\u0026thinsp;0% at 2 weeks after planting (WAP), the Inc decreased to 87.50\u0026thinsp;\u0026plusmn;\u0026thinsp;4.33% at 3 WAP, then increased to 91.67\u0026thinsp;\u0026plusmn;\u0026thinsp;4.87% at 4 WAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). For Sev in cultivar BFM206, it was 2.39\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20 at 2 WAP, decreased to 1.95\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09 at 3 WAP, and increased to 2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 at 4 WAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). In contrast, the Inc and Sev for cultivar BFM260 both decreased over time (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA\u0026amp;B). Indeed, the Inc for BFM260 fell from 68\u0026thinsp;\u0026plusmn;\u0026thinsp;10.95% at 2 WAP to 60\u0026thinsp;\u0026plusmn;\u0026thinsp;12.24% at 3 WAP, and further to 52\u0026thinsp;\u0026plusmn;\u0026thinsp;8.36% at 4 WAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eA). The Sev for BFM260 was 1.78\u0026thinsp;\u0026plusmn;\u0026thinsp;0.10 at 2 WAP, 1.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.12 at 3 WAP, and 1.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08 at 4 WAP (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). The decrease in Inc over time between 2 and 4 WAP was not significant for cultivar BFM206 (P\u0026thinsp;=\u0026thinsp;0.419) and for cultivar BFM260 (P\u0026thinsp;=\u0026thinsp;0.164). For Sev, the reduction between 2 and 4 WAP was not significant for cultivar BFM206 (P\u0026thinsp;=\u0026thinsp;0.162), but the decrease in Sev for cultivar BFM260 from 2 to 4 WAP was significant (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e5.2. Effect of treatment on meristem regeneration rate and callus formation rate\u003c/h2\u003e \u003cp\u003eUnder the heat treatment, the meristem regeneration rates (RRM) observed for the BFM260 and BFM206 cultivars were 60.90\u0026thinsp;\u0026plusmn;\u0026thinsp;12.58% and 34.50\u0026thinsp;\u0026plusmn;\u0026thinsp;10.93%, respectively. The difference in RRM between the cultivars under heat treatment was significant (P\u0026thinsp;=\u0026thinsp;0.008) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). In contrast, the RRM were similar (P\u0026thinsp;=\u0026thinsp;1) between the cultivars under the control treatment, with 75\u0026thinsp;\u0026plusmn;\u0026thinsp;16% for the BFM206 cultivar and 75\u0026thinsp;\u0026plusmn;\u0026thinsp;13.67% for the BFM260 cultivar (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA). Additionally, for the BFM206 cultivar, the RRM under heat treatment differed significantly (P\u0026thinsp;=\u0026thinsp;0.008) from the RRM under the control treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eRegarding the callus formation rate (CFR), it was 44.40\u0026thinsp;\u0026plusmn;\u0026thinsp;15.49% and 46.70\u0026thinsp;\u0026plusmn;\u0026thinsp;7.07% for the BFM206 and BFM260 cultivars, respectively, under heat treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). Under the control treatment, the BFM206 cultivar had an CFR of 50\u0026thinsp;\u0026plusmn;\u0026thinsp;14.43%, and the BFM260 cultivar showed an CFR of 65\u0026thinsp;\u0026plusmn;\u0026thinsp;17.50% (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). The results of the mean separation analysis indicated that the CFR did not vary between the cultivars under heat treatment (P\u0026thinsp;=\u0026thinsp;0.890) or under the control treatment (P\u0026thinsp;=\u0026thinsp;0.337) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB). Two-factor analyse of variance revealed a non-significant interaction between cultivar and treatment for both the RRM (P\u0026thinsp;=\u0026thinsp;0.141) and the CFR (P\u0026thinsp;=\u0026thinsp;0.577) (Table I).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e5.3. Effect of treatment on meristem growth parameters\u003c/h2\u003e \u003cp\u003eUnder the heat treatment, the number of nodes (NN) observed was 3.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 nodes per meristem for cultivar BFM206 and 5.92\u0026thinsp;\u0026plusmn;\u0026thinsp;2.36 nodes per meristem for cultivar BFM260 (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). The difference in the NN between cultivars under the heat treatment was not significant (P\u0026thinsp;=\u0026thinsp;0.303). In the control treatment, cultivar BFM260 had 1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.24 nodes per meristem, and cultivar BFM206 had 1.25\u0026thinsp;\u0026plusmn;\u0026thinsp;2.50 nodes per meristem (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). The difference in the NN between cultivars in the control treatment was also not significant (P\u0026thinsp;=\u0026thinsp;0.998). Additionally, for cultivar BFM260, the NN under the heat treatment was significantly higher (P\u0026thinsp;=\u0026thinsp;0.002) compared to the control treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eFor the number of stems (NS), cultivars BFM206 and BFM260 recorded 1.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 and 2.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60 stems per meristem, respectively, under the heat treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). The mean separation indicated no significant difference (P\u0026thinsp;=\u0026thinsp;0.467) in the NS between the cultivars under the heat treatment. In the control treatment, cultivar BFM206 had 1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1 stems per meristem and cultivar BFM260 had 1.20\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44 stems per meristem (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). The difference in NS between cultivars in the control treatment was also not significant (P\u0026thinsp;=\u0026thinsp;0.990). The results showed that for cultivar BFM260, the NS under the heat treatment was significantly different (P\u0026thinsp;=\u0026thinsp;0.05) from the control treatment.\u003c/p\u003e \u003cp\u003eRegarding the number of roots (NR), cultivars BFM260 and BFM206 developed 1.31\u0026thinsp;\u0026plusmn;\u0026thinsp;2.14 and 0.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 roots per meristem, respectively, under the heat treatment (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC). The mean separation showed no significant difference (P\u0026thinsp;=\u0026thinsp;0.786) in the NR between cultivars under the heat treatment. In the control treatment, no root development was observed for either cultivar (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003eFor the number of leaves (NL), cultivars BFM206 and BFM260 produced 2.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57 and 2.85\u0026thinsp;\u0026plusmn;\u0026thinsp;1.14 leaves per meristem, respectively, under the heat treatment. The difference in the NL between cultivars under the heat treatment was not significant (P\u0026thinsp;=\u0026thinsp;0.994). In the control treatment, cultivar BFM206 had 3.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95 leaves per meristem, and cultivar BFM260 had 2.40\u0026thinsp;\u0026plusmn;\u0026thinsp;1.34 leaves per meristem (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD). The difference in the NL between cultivars in the control treatment was also not significant (P\u0026thinsp;=\u0026thinsp;0.302).\u003c/p\u003e \u003cp\u003eThe two-factor analyse of variance revealed a non-significant cultivar*treatment interaction for the NN (P\u0026thinsp;=\u0026thinsp;0.188), the NS (P\u0026thinsp;=\u0026thinsp;0.424), the NR (P\u0026thinsp;=\u0026thinsp;0.525), and the NL (P\u0026thinsp;=\u0026thinsp;0.156) (Table I).\u003c/p\u003e \u003cp\u003eTable I. Means and two-factor analysis of variance of measured parameters.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Taba\" border=\"1\"\u003e \u003ccolgroup cols=\"7\"\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 \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFactors/Levels\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMRR (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCFR (%)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNN\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eNL\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNR\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCultivars (C)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBFM206\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e45.33\u0026thinsp;\u0026plusmn;\u0026thinsp;15.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e46.80\u0026thinsp;\u0026plusmn;\u0026thinsp;33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.14\u0026thinsp;\u0026plusmn;\u0026thinsp;2.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.14\u0026thinsp;\u0026plusmn;\u0026thinsp;0.37\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eBFM260\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e65.20\u0026thinsp;\u0026plusmn;\u0026thinsp;12.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57.10\u0026thinsp;\u0026plusmn;\u0026thinsp;23.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.56\u0026thinsp;\u0026plusmn;\u0026thinsp;3.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.72\u0026thinsp;\u0026plusmn;\u0026thinsp;1.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.22\u0026thinsp;\u0026plusmn;\u0026thinsp;1.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.94\u0026thinsp;\u0026plusmn;\u0026thinsp;1.89\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP (α\u0026thinsp;=\u0026thinsp;0.05)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.018\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.354\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.039\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.272\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.041\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.525\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatments (T)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eUntreated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75\u0026thinsp;\u0026plusmn;\u0026thinsp;19.76\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57.50\u0026thinsp;\u0026plusmn;\u0026thinsp;24.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;2.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eThermo-treated\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e46.50\u0026thinsp;\u0026plusmn;\u0026thinsp;3.24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e45.20\u0026thinsp;\u0026plusmn;\u0026thinsp;21.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.44\u0026thinsp;\u0026plusmn;\u0026thinsp;2.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.81\u0026thinsp;\u0026plusmn;\u0026thinsp;1.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.44\u0026thinsp;\u0026plusmn;\u0026thinsp;1.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.13\u0026thinsp;\u0026plusmn;\u0026thinsp;1.96\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eP (α\u0026thinsp;=\u0026thinsp;0.05)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.002\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.267\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.546\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.004\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.289\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC*T\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.141\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.577\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.188\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.156\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.424\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.525\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003eMRR: meristem regeneration rate; CFR: callus formation rate; NN: number of nodes; NL: number of leaves; NS: number of stems; NR: number of roots; P: probability; α: significance level; C*T: cultivar*treatment interaction; mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"6. Discussion","content":"\u003cp\u003eThe heat treatment was applied to cuttings infected with cassava mosaic virus. The observed reduction in both incidence and severity of cassava mosaic disease over time on the cuttings after thermal treatment can be explained by the heat-induced reduction in viral activity. Kouassi et al., (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) and Zinga et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) have shown that thermotherapy significantly decreases both the incidence and severity of cassava mosaic disease. Heat limits the replication and movement of viruses within the host (Panattoni et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e\u003csup\u003e)\u003c/sup\u003e leading to a reduction in symptom expression.\u003c/p\u003e \u003cp\u003eThe meristems obtained from shoots both thermo-treated and untreated cuttings were cultured. The significant difference in meristem regeneration rates between the cultivars under heat treatment suggests that the BFM260 meristems exhibit better heat tolerance compared to the BFM206 meristems. Moreover, thermotherapy had a detrimental effect on the regeneration ability of BFM206 meristems compared to the control treatment. These results are consistent with Wang et al. (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), who found that high and prolonged thermal treatments can diminish the regeneration capacity of apices taken from treated shoots. Additionally, Magyar-T\u0026aacute;bori et al. (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) proposed that this reduction in regeneration rate under thermal stress could be due to oxidative damage, which affects cellular metabolism and may lead to cell death. However, the lack of a significant interaction between cultivar and treatment on meristem regeneration indicates that thermotherapy impacts the regeneration of meristems similarly across both cultivars. Therefore, the observed difference in regeneration rates under thermal treatment is more likely related to the inherent regenerative capacity of the meristems of each cultivar rather than their ability to tolerate the thermal treatment.\u003c/p\u003e \u003cp\u003eMeristem regeneration was frequently associated with callus formation. The callus formation rates observed, ranging between 44.40% and 65%, were relatively high. This suggests that the genetic stability of the resulting plantlets needs to be evaluated. Indeed, high callus formation can lead to somaclonal variations, potentially resulting in genetically diverse plantlets (Amitchihou\u0026eacute; et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; George et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2008\u003c/span\u003e)\u003c/p\u003e \u003cp\u003eThe regenerated meristems gave rise to plantlets. The superior performance in terms of the number of nodes and stems observed in the BFM260 cultivar may be attributed to the quality of the culture medium. The medium used appears to be better suited for the growth of BFM260 plantlets compared to those of the BFM206 cultivar. Indeed, different species or varieties have varying requirements for nutrients and growth hormones (Carvalho et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Uwimana et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAdditionally, the higher number of nodes and stems in BFM260 cultivar under heat treatment may also be linked to its low severity of cassava mosaic disease. The reduction in disease severity likely resulted in virus-free meristems, which would have grown better than those from untreated plants. Indeed, during thermal treatment, the replication and movement of viruses to the host\u0026rsquo;s meristematic cells are inhibited by heat (Panattoni et al., \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Wang et al., \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2018\u003c/span\u003e ; Wang et al., \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2008\u003c/span\u003e), thus promoting better in vitro growth. In contrast, untreated plants would produce highly infected meristems, which could lead to reduced growth of plantlets in the control treatment. The inhibition of in vitro growth due to viral infection has been documented in potato (\u003cem\u003eSolanum tuberosum\u003c/em\u003e L.) and apple (\u003cem\u003ePyrus spp\u003c/em\u003e.) (Chen et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Li et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Furthermore, the absence of roots in plantlets from both cultivars under the control treatment may be due to the meristems' inability to utilize auxin present in the culture medium. This inability is likely due to high virus levels in meristems from untreated shoots. Auxin is essential for root formation (Cammarata et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Kurepa \u0026amp; Smalle, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), and its ineffective use and absorption would impair root development (Chen et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Wang et al. (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) also noted that virus-induced difficulty in auxin absorption by plantlets affects normal cellular function, further impeding root development.\u003c/p\u003e"},{"header":"7. Conclusion","content":"\u003cp\u003eThis study evaluated the effects of heat treatment on cassava cuttings in relation to the severity and incidence of cassava mosaic disease, as well as its impact on meristem regeneration and the growth of plantlets derived from meristem cultures. Indeed, the findings revealed that heat treatment of cuttings significantly reduced meristem regeneration rates in the BFM206 cultivar. For the BFM260 cultivar, thermal treatment of cuttings effectively decreased the severity of cassava mosaic disease. Additionally, heat treatment of cuttings enhanced the number of nodes and stems in plantlets of the BFM260 cultivar. Future research should explore optimal culture media to further improve the multiplication and growth of these plantlets.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eBAP\u0026nbsp; \u0026nbsp;\u0026nbsp;6-Benzylamino purine\u003c/p\u003e\n\u003cp\u003eCMD\u0026nbsp; \u0026nbsp;Cassava Mosaic Disease\u003c/p\u003e\n\u003cp\u003eCBSD\u0026nbsp;Cassava Brown Streak Disease\u003c/p\u003e\n\u003cp\u003eMS\u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Murashige and Skoog medium\u003c/p\u003e\n\u003cp\u003eNAA \u0026nbsp; 1-Naphthaleneacetic acid\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eConflicts of interests\u003c/h2\u003e \u003cp\u003eThe authors declare that there are no conflicts of interests regarding the publication of this article.\u003c/p\u003e\u003ch2\u003eAcknowledgements\u003c/h2\u003e \u003cp\u003eThe authors are grateful to the \u0026ldquo;Minist\u0026egrave;re de l\u0026rsquo;Enseignement Sup\u0026eacute;rieur, de la Recherche et de l\u0026rsquo;Innovation (MESRI)\u0026rdquo; through the \u0026ldquo;Programme d\u0026rsquo;Appui \u0026agrave; l\u0026rsquo;Enseignement Sup\u0026eacute;rieur (PAES)\u0026rdquo; for funding this work. We also wish to thank the \u0026ldquo;Institute de l\u0026rsquo;Environnement et de la Recherche Agricole (INERA)\u0026rdquo; for providing the plant material and hosting this work.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAmitchihou\u0026eacute; FS, Jane WK, Elijah A, Jerome AH, Colombe D, Peter N, Corneille A (2019) Callus induction in three mosaic disease resistant cassava cultivars in Benin and genetic stability of the induced calli using simple sequence repeat (SSR) and sequence-characterized amplified region (SCAR) markers. Afr J Biotechnol 18:1044\u0026ndash;1053. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5897/ajb2019.16967\u003c/span\u003e\u003cspan address=\"10.5897/ajb2019.16967\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBenke AP, Krishna R, Khandagale K, Gawande S, Shelke P, Dukare S, Dhumal S, Singh M, Mahajan V (2023) Efficient elimination of viruses from garlic using a combination of shoot meristem culture, thermotherapy, and chemical treatment. Pathogens 12. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/pathogens12010129\u003c/span\u003e\u003cspan address=\"10.3390/pathogens12010129\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCammarata J, Roeder AHK, Scanlon MJ (2023) The ratio of auxin to cytokinin controls leaf development and meristem initiation in \u003cem\u003ePhyscomitrium patens\u003c/em\u003e. J Exp Bot 74:6541\u0026ndash;6550. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/jxb/erad299\u003c/span\u003e\u003cspan address=\"10.1093/jxb/erad299\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCampos H, Caligari PDS (2017) Chapter 5: Cassava (\u003cem\u003eManihot esculenta\u003c/em\u003e Crantz) in genetic improvement of tropical crops. In Genetic Improvement of Tropical Crops. 1\u0026ndash;52. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/978-3-319-59819-2\u003c/span\u003e\u003cspan address=\"10.1007/978-3-319-59819-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCarvalho MJS, Oliveira EJ, Souza AS, Pereira JS, Diamantino MSAS, Oliveira SAS (2017) Cleaning cassava genotypes infected with cassava frogskin disease via \u003cem\u003ein vitro\u003c/em\u003e shoot tip culture. Genet Mol Res 16:1\u0026ndash;17. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4238/gmr16029556\u003c/span\u003e\u003cspan address=\"10.4238/gmr16029556\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen J, Tang HH, Li L, Qin SJ, Wang GP, Hong N (2017) Effects of virus infection on plant growth, root development and phytohormone levels in in vitro-cultured pear plants. Planr Cell Tissue Organ Cult 131:359\u0026ndash;368. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s11240-017-1289-1\u003c/span\u003e\u003cspan address=\"10.1007/s11240-017-1289-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChikoti PC, Mulenga RM, Tembo M, Sseruwagi P (2019) Cassava mosaic disease: a review of a threat to cassava production in Zambia. J Plant Pathol 101:479\u0026ndash;479. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s42161-019-00267-w\u003c/span\u003e\u003cspan address=\"10.1007/s42161-019-00267-w\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDabir\u0026eacute; R, Belem J (2003) Les plantes \u0026agrave; tubercules et racines au Burkina Faso. Wasnet 8:12\u0026ndash;16\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEl-Sharkawy MA (2004) Cassava biology and physiology. Plant Mol Biol 56:481\u0026ndash;501. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s11103-005-2270-7\u003c/span\u003e\u003cspan address=\"10.1007/s11103-005-2270-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFAO (2022) Food and Agriculture Organization of the United Nations. FAOSTAT statistics database, Rome\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFerguson M, Choti A, Munguti F (2020) Virus Elimination and Testing in Cassava: A pratical manual. IITA, Ibadan, Nigeria. : 1\u0026ndash;24\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGeorge E, Hall M, De Klerk J (2008) Plant propagation by tissue culture 3rd edition Volume 1. The backgorund\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGraciela MB, Roa CJE, Aranzales ER, Debouck DG (2010) Handbook of procedures for \u003cem\u003ein vitro\u003c/em\u003e germplasm conservation\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHu G jun, Dong Y, feng, Zhang Z ping, Fan X, dong, Ren F (2021) Elimination of \u003cem\u003eGrapevine fleck virus\u003c/em\u003e and \u003cem\u003eGrapevine rupestris stem pitting-associated virus\u003c/em\u003e from \u003cem\u003eVitis vinifera\u003c/em\u003e 87\u0026thinsp;\u0026ndash;\u0026thinsp;1 by ribavirin combined with thermotherapy. Journal of Integrative Agriculture 20: 2463\u0026ndash;2470. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/S2095-3119(20)63336-6\u003c/span\u003e\u003cspan address=\"10.1016/S2095-3119(20)63336-6\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eINSD (2019) Cinqui\u0026egrave;me recensement g\u0026eacute;n\u0026eacute;ral de la population et de l\u0026rsquo;habitation du Burkina Faso. synth\u0026egrave;se des r\u0026eacute;sultats d\u0026eacute;finitifs\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKouassi MK, Kouassi N, Yeo FE (2019) Caract\u0026eacute;ristiques d\u0026rsquo;un dispositif de thermoth\u0026eacute;rapie construit localement pour l\u0026rsquo;assainissement de plants de manioc. AISA D\u0026eacute;veloppement n\u0026deg; 16:2\u0026ndash;5. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.13140/RG.2.2.14034.53448\u003c/span\u003e\u003cspan address=\"10.13140/RG.2.2.14034.53448\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKrishna R, Ansari WA, Khandagale K, Benke AP, Soumia PS, Manjunathagowda DC, Gawande SJ, Ade AB, Mokat DN, Singh M (2022) Meristem culture: a potential technique for \u003cem\u003ein vitro\u003c/em\u003e virus-free plants production in vegetatively propagated crops. Advances in Plant Tissue Culture. Elsevier, pp 325\u0026ndash;343. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/B978-0-323-90795-8.00017-5\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-323-90795-8.00017-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKurepa J, Smalle JA (2022) Auxin/Cytokinin antagonistic control of the shoot/root growth ratio and its relevance for adaptation to drought and nutrient deficiency stresses. Int J Mol Sci 23. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/ijms23041933\u003c/span\u003e\u003cspan address=\"10.3390/ijms23041933\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLegg J, Winter S (2020) Cassava mosaic viruses (\u003cem\u003eGeminiviridae\u003c/em\u003e). Encyclopedia of virology. Elsevier, pp 301\u0026ndash;312. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/B978-0-12-809633-8.21523-9\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-12-809633-8.21523-9\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLegg JP, Lava Kumar P, Makeshkumar T, Tripathi L, Ferguson M, Kanju E, Ntawuruhunga P, Cuellar W (2014) Cassava virus diseases. Advances in Virus Research. Elsevier Inc., pp 85\u0026ndash;142. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/bs.aivir.2014.10.001\u003c/span\u003e\u003cspan address=\"10.1016/bs.aivir.2014.10.001\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi JW, Wang B, Song XM, Wang RR, Chen L, Zhang H, Zhang ZB, Wang QC (2013) \u003cem\u003ePotato leafroll virus (PLRV)\u003c/em\u003e and \u003cem\u003ePotato virus Y (PVY)\u003c/em\u003e influence vegetative growth, physiological metabolism, and microtuber production of in vitro-grown shoots of potato (\u003cem\u003eSolanum tuberosum\u003c/em\u003e L.). Plant Cell, Tissue and Organ Culture 114: 313\u0026ndash;324. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s11240-013-0327-x\u003c/span\u003e\u003cspan address=\"10.1007/s11240-013-0327-x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMAAH (2020) Rapport de l\u0026rsquo;enqu\u0026ecirc;te nationale int\u0026eacute;gr\u0026eacute; de s\u0026eacute;curit\u0026eacute; alimentaire et nutritionnelle, Burkina Faso\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMagyar-T\u0026aacute;bori K, Mendler-Drienyovszki N, Han\u0026aacute;sz A, Zsombik L, Dobr\u0026aacute;nszki J (2021) Phytotoxicity and other adverse effects on the in vitro shoot cultures caused by virus elimination treatments: reasons and solutions. Plants 10:670. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/plants10040670\u003c/span\u003e\u003cspan address=\"10.3390/plants10040670\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePlan manioc (2019) Plan d\u0026rsquo;actions pour le d\u0026eacute;veloppement de la Fili\u0026egrave;re. manioc du Burkina Faso\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMontagnac JA, Davis CR, Tanumihardjo SA (2009) Nutritional value of cassava for use as a staple food and recent advances for improvement. Compr Rev Food Sci Food Saf 8:181\u0026ndash;194. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1541-4337.2009.00077.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1541-4337.2009.00077.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMurashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473\u0026ndash;497\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNakabonge G, Nangonzi R, Tumwebaze BS, Kazibwe A, Samukoya C, Baguma Y (2020) Production of virus-free cassava through hot water therapy and two rounds of meristem tip culture. Cogent Food Agric 6:1800923. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1080/23311932.2020.1800923\u003c/span\u003e\u003cspan address=\"10.1080/23311932.2020.1800923\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNehra NS, Kartha KK (1994) Meristem and shoot tip culture: requirements and applications. Plant cell and tissue culture. Dordrecht, Springer Netherlands, pp 37\u0026ndash;70. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/978-94-017-2681-8_3\u003c/span\u003e\u003cspan address=\"10.1007/978-94-017-2681-8_3\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOlutosin AO, Sawicka B (2019) Cassava, a 21st century staple crop: how can Nigeria harness its enormous trade potentials? Acta Sci Agric 3:194\u0026ndash;202. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.31080/asag.2019.03.0586\u003c/span\u003e\u003cspan address=\"10.31080/asag.2019.03.0586\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOuedraogo A (2023) Ins\u0026eacute;curit\u0026eacute; alimentaire et r\u0026eacute;silience au Burkina Faso: une analyse compar\u0026eacute;e des m\u0026eacute;nages des personnes d\u0026eacute;plac\u0026eacute;es internes (pdi) des centres d\u0026rsquo;accueil et des m\u0026eacute;nages locaux dans la commune\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePanattoni A, Luvisi A, Triolo E (2013) Review. Elimination of viruses in plants: twenty years of progress. Span J Agricultural Res 11:173\u0026ndash;188. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5424/sjar/2013111-3201\u003c/span\u003e\u003cspan address=\"10.5424/sjar/2013111-3201\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSaranraj P, Behera SS, Ray RC (2019) Chapter 7 - Traditional foods from tropical root and tuber crops: innovations and challenges. Innovations in traditional foods, Galanakis CMBT-I in TF. Woodhead Publishing, pp 159\u0026ndash;191. DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/B978-0-12-814887-7.00007-1\u003c/span\u003e\u003cspan address=\"10.1016/B978-0-12-814887-7.00007-1\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShirima RR, Maeda DG, Kanju EE, Tumwegamire S, Ceasar G, Mushi E, Sichalwe C, Mtunda K, Mkamilo G, Legg JP (2019) Assessing the degeneration of cassava under high-virus inoculum conditions in Coastal Tanzania. Plant Dis 103:2652\u0026ndash;2664. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1094/PDIS-05-18-0750-RE\u003c/span\u003e\u003cspan address=\"10.1094/PDIS-05-18-0750-RE\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSoro M, Tiendr\u0026eacute;b\u0026eacute;ogo F, Pita JS, Traor\u0026eacute; ET, Som\u0026eacute; K, Tibiri EB, N\u0026eacute;ya JB, Mutuku JM, Simpor\u0026eacute; J, Kon\u0026eacute; D (2021) Epidemiological assessment of cassava mosaic disease in Burkina Faso. Plant Pathol 70:2207\u0026ndash;2216. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/ppa.13459\u003c/span\u003e\u003cspan address=\"10.1111/ppa.13459\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSu YH, Liu YB, Zhang XS (2011) Auxin-cytokinin interaction regulates meristem development. Mol Plant 4:616\u0026ndash;625. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/mp/ssr007\u003c/span\u003e\u003cspan address=\"10.1093/mp/ssr007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTiendr\u0026eacute;b\u0026eacute;ogo F, Lefeuvre P, Hoareau M, Traor\u0026eacute; VSE, Barro N, Reynaud B, Traor\u0026eacute; AS, Konat\u0026eacute; G, Traor\u0026eacute; O, Lett J-M (2009) Occurrence of \u003cem\u003eEast African cassava mosaic virus‐Uganda (EACMV‐UG)\u003c/em\u003e in Burkina Faso. Plant Pathol 58:783\u0026ndash;783. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1365-3059.2009.02110.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1365-3059.2009.02110.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eUwimana J, Sakha M, Danga B, Gitari H, Gweyi-Onyango JP (2022) Involvement of plant growth regulators and varieties in the multiplication of cassava planting materials: a case study of Rwanda. Int J Bioresource Sci 9. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.30954/2347-9655.02.2022.14\u003c/span\u003e\u003cspan address=\"10.30954/2347-9655.02.2022.14\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang CX, Hong N, Wang GP, Jiang B, Fan XD (2009) Effects of c\u003cem\u003eitrus tristeza virus\u003c/em\u003e on the growth of in vitro-cultured citrus. J Plant Pathol 91:357\u0026ndash;363\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang MR, Cui ZH, Li JW, Hao XY, Zhao L, Wang QC (2018) \u003cem\u003evitro\u003c/em\u003e thermotherapy-based methods for plant virus eradication. Plant Methods 14:1\u0026ndash;18. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/s13007-018-0355-y\u003c/span\u003e\u003cspan address=\"10.1186/s13007-018-0355-y\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang Q, Cuellar WJ, Rajam\u0026auml;ki ML, Hirata Y, Valkonen JPT (2008) Combined thermotherapy and cryotherapy for efficient virus eradication: relation of virus distribution, subcellular changes, cell survival and viral RNA degradation in shoot tips. Mol Plant Pathol 9:237\u0026ndash;250. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1364-3703.2007.00456.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1364-3703.2007.00456.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eY\u0026eacute;o EF, Kouassi MK, Pita JS, Kouassi NK, Kon\u0026eacute; D, N\u0026rsquo;guetta SPA (2020) Using thermotherapy and meristem tip culture for producing virus-free cassava planting material from six varieties cultivated in C\u0026ocirc;te d\u0026rsquo;ivoire. Int J Sci Technol Res 9:1607\u0026ndash;1612\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZinga I et al (2014) Field evaluation of the effectiveness of thermotherapy against \u003cem\u003ecassava mosaic disease\u003c/em\u003e in Central African Republic. Am J Experimental Agric 4:1232\u0026ndash;1241. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.9734/ajea/2014/10275\u003c/span\u003e\u003cspan address=\"10.9734/ajea/2014/10275\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"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":"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":"Manihot esculenta, thermotherapy, meristem, regeneration, growth","lastPublishedDoi":"10.21203/rs.3.rs-4969011/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4969011/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eUsing healthy cuttings can enhance the productivity of cultivars susceptible to viral diseases. However, due to the unavailability of healthy cuttings, farmers often have to rely on infected ones. This study aims to enhance the health quality of planting material by using thermotherapy and meristem culture techniques on two cassava cultivars. To achieve this, cuttings from the BFM206 and BFM260 cultivars were subjected to heat treatment in a thermotherapy chamber and a control treatment in field conditions. The heat treatment involved exposing the cuttings to a temperature of 37\u0026deg;C for the first week. The temperature was then increased by 2\u0026deg;C each week over four weeks, reaching 37\u0026deg;C, 39\u0026deg;C, 41\u0026deg;C, and 43\u0026deg;C, respectively. After the treatment, the resulting shoots were used as mother plants for meristem culture. The experimental design was a split-plot with 5 repetitions. The parameters studied included the severity and incidence of cassava mosaic disease, the regeneration rate, the callus formation rate, and growth parameters. The heat treatment resulted in a significant difference between the cultivars in terms of the severity of cassava mosaic disease (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and the regeneration rate (P\u0026thinsp;=\u0026thinsp;0.008). Furthermore, compared to the control treatment, the heat treatment significantly increased the number of nodes by 4.92 and the number of stems by 1.42 in the BFM260 cultivar. In conclusion, thermotherapy can help reduce the severity of cassava mosaic disease in mother plants and thereby improve the in vitro growth of meristems.\u003c/p\u003e","manuscriptTitle":"Effect of thermotherapy on meristem culture response of two local cassava (Manihot esculenta Crantz) cultivars from Burkina Faso","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-08-27 10:28:31","doi":"10.21203/rs.3.rs-4969011/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":"3eab3b11-aaaa-4c24-94de-67aac1de5b7d","owner":[],"postedDate":"August 27th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":36520947,"name":"Agronomy"},{"id":36520948,"name":"Biotechnology and Bioengineering"},{"id":36520949,"name":"Food Science \u0026 Technology"}],"tags":[],"updatedAt":"2024-08-27T10:28:31+00:00","versionOfRecord":[],"versionCreatedAt":"2024-08-27 10:28:31","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4969011","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4969011","identity":"rs-4969011","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}