Evaluation of four growth mediums effect on potato (Solanum tubérosum L.) plantlet production in Burkina Faso

Evaluation of four growth mediums effect on potato (Solanum tubérosum L.) plantlet production in 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 Evaluation of four growth mediums effect on potato (Solanum tubérosum L.) plantlet production in Burkina Faso Afoussatou ZERBO, Koussao SOME, Djakaridja TIAMA, Monique SORO, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4200930/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 In Burkina Faso, potato growers are faced with difficulties in obtaining seed. These seeds, mostly imported partially meet the quality requirement in potato production. Furthermore, these seeds are not accessible in term of cost and also not available in time. Local seed production is therefore necessary. In vitro culture can remedy this problem by providing quality seeds. Thus, the goal of our study was to set up an in vitro culture protocol for the production of plantlets with optimal vegetative growth. This study was realised in the in vitro culture laboratory at INERA Kamboinsé. Explants from in vitro plantlets (1 cm) of the same generation were used as plant material and transplanted onto four different mediums. The mediums used were: M0= 4,4g/l MS medium (Murashige and Skoog 1962) + 7g/l agar; M1= M0 + 30g/l table sugar; M2= M0 + 0,5mg/l AIB and M3= M0+30g/l sugar + 0,5mg/l AIB. These mediums were prepared with a pH=5,7±0.1, then divided into jars and autoclaved for 20 minutes at a pressure of 120 bars. Subculture in the jars was realised out in a laminar flow hood. The covered jars were transferred to a growth room for 30 days. The results of this study indicated that M1 and M3 showed good regeneration of the in vitro plantlets compared to M0 and M2. Sugar probably had a positive effect on root length, stem diameter, number of nodes and number of leaves open. These parameters strongly differentiated M1 from the other mediums. AIB had a positive effect on the root proliferation of in vitro plantlets in the M2 medium. The combined effect of sugar and AIB was even more positive on stem height, number of roots, number of leaves open and weight of in vitro plantlets, which strongly differentiated medium M3 from the other mediums. In conclusion, M3 proved to be the best medium for producing in vitro plantlets with good vegetative growth for seed production. Micropropagation sugar auxin AIB in vitro plantlets potato and Burkina Faso Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Seeds are essential in most agricultural production systems and for farmers. Since the Green Revolution of the 1960s, the use of improved seeds have played an important role in the development of agriculture [1]. Improved seed has been qualified as the engine of agricultural progress through its ability to increase productivity, improve incomes and food security [1;2]. According to [1] seed technology is a set of applied sciences, technologies and socio-economic aspects that contribute to the production and availability of good quality seed for sowing. The potato seed is not excluded from this technology. Potato is considered the fourth most important food crop in the world after maize, rice and wheat [3]. Globally, over the last 20 years, potato production has increased at a much higher rate in Asia, Africa and Latin America, rising from less than 30 million tonnes in the early 1960s to more than 165 million tonnes in 2007 [4]. Half of its current global production is in China and India [4].The observed increase is thought to be due to the use of in vitro cultivation technology, which is used worldwide for seed production. According to [5], this technology has been used to produce disease-free seeds. Potato seed can be produced via in vitro propagation in two ways, after micropropagation of the explants. The in vitro plantlets thus obtained can be used, after acclimatisation, to produce minitubers, or they can be induced to produce microtubers. The use of in vitro plantlets or microtubers for further seed production can also be achieved by several methods, such as hydroponics, aeroponics and many others. According to [6], aeroponics as promoted by the International Potato Center (CIP), has several advantages, including high multiplication rates. This methodology is also used in sub-Saharan Africa including Burundi, Ethiopia, Kenya, Malawi, Rwanda, Tanzania and Uganda and has led to a large increase in minitubers production although this increase is still small [7;8]. Despite this increase in potato production, the majority of developing countries, particularly those in West Africa, are faced difficulties in obtaining seed [10;11]. Burkina Faso is not an exception to this constraint, potato growers are faced with difficulties in obtaining seed [12]. According to statistics from the Ministry of Agriculture, Animal Resources and Fisheries, most of the seeds used by producers come from outside the country. In fact, two types of potato seed are used in Burkina Faso: farm- seed from previous production and certified seed imported from outside the country, especially from Europe. Imported seed, which are mainly used, only partially meets the required quality criteria and is not available in time [7;12]. As a result, local seed production needs to be developed. In vitro culture technology through the micropropagation of vitroplants can help to alleviate this problem by providing growers with quality seeds. This technique requires the use of appropriate culture mediums. Various culture mediums composed of MS medium (Murashige and skoog 1962), sugar and auxin AIB have been tested. MS medium is generally used as a basic medium in many in vitro culture [13;14;15]. A study by [16] showed the effectiveness of the MS medium (Murashige and skoog 1962) on the development of somatic embryos from the. picholine marocaine ( Olea europaea L ). Sucrose is the most effective source of carbohydrate [17]. According to [18], the use of table sugar has improved potato micropropagation but has also reduced production costs by 34 to 51% compared to analytical grade sucrose. Auxin is a growth hormone and is required for pollen embryo induction in many Solanaceae species except tobacco [19]. Hence our interest in using these elements. The purpose of our study was to identify a culture medium suitable for micropropagation of potato in vitro plantlets in Burkina Faso. Specifically, in combination with the MS base medium, we aimed to: i) evaluate the effect of sugar on the development of in vitro plantlets; ii) evaluate the effect of the auxin indole-3-butyric acid (IBA) on the development of in vitro plantlets; and iii) evaluate the combined effect of sugar and the auxin IBA on in vitro plantlets. Materials And Methods Material The plant material is consisted of tubers of improved potato varieties and in vito plantlet genotypes from the International Potato Center (CIP). A total of two improved varieties ( Sahel and Spunta ) and eight CIP genotypes were used for in vitro culture. Methodology The work was carried out at INERA station of Kamboinsé, in the in vitro culture laboratory. The tubers of the improved varieties underwent initiation, then multiplication before being used as plant material. The axillary buds of approximately 1 cm, consisting of a node of these in vitro plantlets, were used as explants and transplanted onto each of the four cultures mediums at a rate of 10 ml of culture medium per explant. The culture medium consisted of : M0= 4,4g/l MS (Murashige and skoog 1962) + 7g/l agar, M0 is the base medium; M1= M0 + 30g/l sugar; M2= M0 + 0,5mg/l auxin 3 indole butyric acid (IBA) and M3= M0+30g/l sugar + 0,5mg/l auxin IBA. The pH of each medium was adjusted to pH=5.7±0.1 with 1N NaOH. A 909 ml cylindrical glass jar was used as the culture vessel. The prepared media were dispensed into the containers and then autoclaved for 20 minutes at a pressure of 120 bar before subculturing in the jars under a laminar flow hood. These containers containing the explants were loosely covered and placed in a culture room for 30 days at a temperature of 22 ± 1 o C, 70% relative humidity, a photoperiod of 16 h and a lighting intensity of 1200 lux. Statistical analysis Excel 2016 was used for data entry. XLSTAT 2016 was used to perform an analysis of variance of the data to determine the effect of the different environments on the growth parameters of the plantlets. The results of this analysis were used to produce tables, frequency calculations and histograms using Excel 2016. Results Regeneration rate of plantlets Table1 Analysis of variance of environmental data Media Growth parameters M0 M1 M2 M3 Pr > F NRE 0,092 a 0,817 b 0,067 a 0,767 b 0,000 HTI 0,061 a 1,905 b 0,025 a 2,584 c 0,000 DTI 0,007 a 0,076 b 0,003 a 0,071 b 0,000 NFE 0,033 a 0,625 b 0,000 a 1,675 c 0,000 LFE 0,002 a 0,025 b 0,000 a 0,044 c 0,000 NNO 0,158 a 5,000 b 0,067 a 4,846 b 0,000 DEN 0,010 a 0,371 b 0,000 a 0,529 c 0,000 NRM 0,000 a 0,321 b 0,000 a 0,571 c 0,000 NOR 0,017 a 1,783 b 0,117 a 4,321 c 0,000 LRA 0,002 a 0,371 c 0,001 a 0,304 b 0,000 PPD 0,001 a 0,016 b 0,001 a 0,026 c 0,000 NRA 0,025 a 0,254 b 0,000 a 0,233 b 0,000 NRE: Number of shoots; HTI: Height of stem (cm), DTI: Diameter of stem (mm), NFE: Number of leaves, LFE: Length of leaf (cm), NNO: Number of nodes, DEN: Distance between nodes (cm), NRM: Number of branches, NOR: Number of roots, LRA: Length of roots (cm), PPD: Weight of in vitro plants (g), NRA: Number of aerial roots. The result of the analysis of variance showed a very highly significant difference between the growth parameters of the different media with a Pr > F (0,0001) (Table 1). Statistical analysis also showed that all the media used favoured vegetative regeneration of in vitro plantlets, with the rate varying from one medium to another. As a result, mediums M1 and M3 showed the best regeneration rates, ranging from 46,89% to 44,02% respectively. However, low regeneration rates were observed in media M0 and M2, at 5,26% and 3,83% respectively ( Fig. 1 ). Effect of sugar on the development of in vitro plantlets The results showed that sugar had a positive effect on 12 growth parameters measured. The parameters most affected were: root length, stem diameter, number of nodes and number of leaves open. Root length varied from 0,05 to 2,2cm. Long roots of more than 0,5 cm were observed in medium M1, with a frequency of around 63,87% of in vitro plants. The diameter of the plantlets varied from 0,5 to 4 mm. The majority of plantlets (81,04%) had a diameter of between 0,05 and 0.1 cm and were observed in the M1 medium. The number of nodes and the number of opened leaves varied from 1 to 14 nodes and from 1 to 7 leaves, respectively. The majority of plantlets with a high node number of more than 5 and a number of opened leaves of between 1 and 4 were observed in medium M1, with a frequency of 72,21% and 79,75% of plantlets respectively ( Fig 2 ). Effect of auxin indole-3-butyric acid (IBA) on the development of in vitro plantlets This analysis showed that auxin indole-3-butyric acid (IBA) had a positive effect on root proliferation and calogenesis. In fact, we observed root proliferation in medium M2 with 5,83% of in vitro plantlets (Fig.3). Combined effect of sugar and auxin indole-3-butyric acid (IBA) on the development of in vitro plantlets This analysis shows that the combined effect of sugar and auxin AIB was positive on in vitro plant growth parameters. The parameters most affected were: stem height, number of roots, number of leaves open and weight of in vitro plantlets, which strongly differentiated M3 from the other media. The height of the plantlets varied from 0,1 to 8 cm. In fact, 29,67% of in vitro plantlets with a height of between 4 and 6 cm were observed in medium M3. The number of roots varied from 1 to 15, with 46,22% of the plantlets in medium M3 having between 6 and 15 roots. The number of opened leaves varied from 1 to 12 leaves, 46,61% of the plantlets in medium M3 had an opened leaves number between 1 and 4. The weight of the plantlets varied from 0,003 to 0,1g, 41,34% of the M3 plantlets had a weight of between 0,003 and 0,05g (Fig. 4). Discussion The low regeneration of in vitro plantlets in mediums M0 and M2 could be due to a lack of sugar in both mediums. Sugar would be the missing element in these media compared to mediums M1 and M3, which showed good regeneration (more than 5%). In this sense [20] showed that sugar is an essential element for both the vegetative development of cuttings and microtuberisation. According to these authors, the presence of sugar in a growing medium is favorable to cauline and root growth. Our results showed a positive effect of sugar in the M1 medium on growth parameters. The number of nodes, number of leaves open, stem diameter and root length were the parameters most influenced by the effect of sugar. The 30 g/l sugar dose of the M1 medium produced in vitro plantlets with more than 5 nodes and a number of open leaves of between 1 and 4, with a stem diameter of around 1mm and a root length of more than 0,5 cm. We can therefore say that sugar at this dose favours the stem and root development of in vitro plantlets. According to [18], table sugar improves potato micropropagation. Sugar is therefore a source of energy used by explants for their growth and development. The same applies to exogenous sucrose, which is thought to be directly involved in regulating cell proliferation, photosynthesis and the defence mechanism against reactive oxygen derivatives [13]. According to [21] sucrose is the best source of carbon in Ipomoea batatas L , followed by fructose and glucose. Furthermore, [14], have shown that sucrose, glucose and maltose have a favourable effect on various growth parameters in potatoes. We can therefore say that in vitro multiplication of vegetative explants would require a carbon source such as sugar. The root development of in vitro plantlets in the M2 medium is thought to be due to the effect of auxin AIB. Auxin is thought to be produced in the apical buds of plants and transported from stems to roots via a polar transport system [22]. It is accumulated in the root tips and stimulates rhizogenesis by favouring the initiation and development of lateral roots [22]. According to [23;24], AIB positively improves the rooting phase in in vitro culture in cork oak and apple. [21] showed that auxin AIB gave the best rooting parameters compared to ANA for root formation in Ipomoea batatas L . Auxin is known to regulate many aspects of plant growth and development, such as vascular tissue differentiation, embryonic development, root formation and primary and secondary stem formation [25]. Explant regeneration was promoted by the effect of sugar. This regeneration is essential for the production and assimilation of auxin in vitro plantlets. As a result, the combination of sugar and auxin will accentuate the action of auxin through high root production and will allow good development of the in vitro plantlets observed in the M3 medium compared to the other mediums. Indeed, [26;27] reported that the application of glucose, sucrose and fructose stimulated the development of seedlings of Dactylorhiza species and Physocarpus opulifolius (L.) and that the growth rate and root length of seedlings increased in the presence of AIB and α-naphthalene-acetic acid. Thus, the combined effect of sugar and auxin AIB allows the regeneration of explants and the growth of their roots. The in vitro plantlets in medium M3 were characterised by an average height, a higher number of roots, a higher number of leaves and a higher weight of plantlets compared to those in medium. These characteristics would be very important for good acclimatisation of the plantlets. Indeed [21] showed that the better rooting rate observed with AIB favoured 100% success at the acclimatisation stage in sweet potato ( Ipomoea batatas L) . Conclusion This study revealed that the four media used favour the regeneration of in vitro plantlets, but M1 and M3 are the best mediums for good vegetative regeneration. Sugar had a positive effect on growth parameters. Sugar favoured the development of stem diameter, root length, number of nodes and fuller leaves. AIB auxin had a positive effect on root proliferation of in vitro plantlets. The combination of sugar and auxin AIB improved the development of growth parameters such as stem height, number of roots, number of expanded leaves and the weight of in vitro plantlets in medium M3 compared with the other mediums. Thus, the production of plantlets showing good vegetative growth would allow good acclimatisation an improvement of the production of potato seeds. Declarations Availability of data The data used to support the results of this study will be available on request from the corresponding author. Conflicts of interest The authors declare that there are no conflicts of interest in the publication of this article. Acknowledgements The authors would like to thank the members of the Plant Genetics and Improvement team of the Biosciences Laboratory of University Joseph KI-ZERBO and the Institute of the Environment and Agricultural Research (INERA) Kamboinsé, more specifically the members of the in vitro culture laboratory, for their collaboration in carrying out this study. They would also like to thank Dr Idrissa OUEDRAOGO of the Centre Universitaire de Dori / Université Thomas SANKARA for his advice and suggestions. Ethics declarations A licence has been obtained for the use of the plant material. The study complies with national guidelines. References M. Turner, Les semences Quae Cta Presses agronomiques de Gembloux. 2010. M. 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Katarzyna, and J. Ewelina, “The effect of carbon source in culture medium on micropropagation of common ninebark ( Physocarpus opulifolius ( L .) Maxim .) ‘ Diable D ’ or ’ Agnieszka Ilczuk , Katarzyna Jagie łł o-Kubiec , Ewelina Jacygrad,” vol. 12, no. 3, pp. 23–33, 2013. Additional Declarations No competing interests reported. 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-4200930","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":289648067,"identity":"378ab35a-e97b-47ed-805a-10f66558a8e8","order_by":0,"name":"Afoussatou ZERBO","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5ElEQVRIiWNgGAWjYDAC5gNwJuMDBoYDuNQhAbYEMCUB0m1AshY2CaK08LcxH5P88ceujl8i/Vk1T80dOX4G5oePbuDRInGMLU2aty1ZQnJGQtptnmPPjCUb2IyNc/BZc7/HTJqxgVnC4EbCsds8bIcTNxzgYZPGp0X+GI8Z0GH1QC2JbcU8/4jQYgDUIgE0HKglmY2Zt40ILYbH2JKteduOS87secYsObfvsLFkMwG/yB1jPnjzx59qfn729Icf3nw7LMfP3vzwMV7vIwMmHhDJTKxyEGD8QYrqUTAKRsEoGDEAACqWSLqSczNGAAAAAElFTkSuQmCC","orcid":"","institution":"Université Joseph KI-ZERBO","correspondingAuthor":true,"prefix":"","firstName":"Afoussatou","middleName":"","lastName":"ZERBO","suffix":""},{"id":289648068,"identity":"4e3c7eb4-9abc-41ed-87f2-79b2d3f4652e","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":289648069,"identity":"99f74f7e-d433-40b2-acc8-ae22e734ec06","order_by":2,"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":289648070,"identity":"7fe8e3eb-fcd0-4264-8db9-b46c9a834075","order_by":3,"name":"Monique SORO","email":"","orcid":"","institution":"Institut de l’Environnement et de Recherches Agricoles (INERA)","correspondingAuthor":false,"prefix":"","firstName":"Monique","middleName":"","lastName":"SORO","suffix":""},{"id":289648071,"identity":"4c71a8e3-50e2-49a1-830e-321d8f0522f3","order_by":4,"name":"Wendpang-yidé Idrissa Caleb OUEDRAOGO","email":"","orcid":"","institution":"Université Joseph KI-ZERBO","correspondingAuthor":false,"prefix":"","firstName":"Wendpang-yidé","middleName":"Idrissa Caleb","lastName":"OUEDRAOGO","suffix":""},{"id":289648072,"identity":"3b61f19a-5b96-4d50-ae5d-423faca31403","order_by":5,"name":"Renan Ernest TRAORE","email":"","orcid":"","institution":"Université Joseph KI-ZERBO","correspondingAuthor":false,"prefix":"","firstName":"Renan","middleName":"Ernest","lastName":"TRAORE","suffix":""}],"badges":[],"createdAt":"2024-04-01 13:29:30","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4200930/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4200930/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":54515239,"identity":"db298271-677f-4e3d-8f31-c49e738f93ee","added_by":"auto","created_at":"2024-04-11 16:24:19","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":32453,"visible":true,"origin":"","legend":"\u003cp\u003eRegeneration rate of the in vitro plantlets by culture medium (M0= MS (Murashige and skoog 1962) 4,4g/l + agar 7g/l ; M1= M0 + 30g/l sugar ; M2= M0 + 0,5mg/l auxin 3 indole butyric acid (IBA) et M3= M0+30g/l de sugar + 0,5mg/l auxin 3 indole butyric acid IBA)\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-4200930/v1/00b7321adecd49796f59da93.png"},{"id":54515238,"identity":"126763dd-b1ff-4675-b1b3-276fc319d1b0","added_by":"auto","created_at":"2024-04-11 16:24:19","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":63465,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of sugar on in vitro plantlet growth parameters: a) root length, b) stem diameter, c) number of nodes, and d) number of fully expanded leaves (Murashige and skoog 1962) 4,4g/l + 7g/l agar = M0; M1= M0 + 30g/l sugar)\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-4200930/v1/b1f969f4d6c20e77f3f4c5db.png"},{"id":54515243,"identity":"cbcf6053-bc03-438b-affd-a1cb94c6e3ab","added_by":"auto","created_at":"2024-04-11 16:24:20","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":491949,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of auxin AIB on root development of in vitro plantlets. (Murashige and skoog 1962) 4,4g/l + 7g/l agar; M1= M0 + 30g/l sugar; M2= M0 + 0,5mg/l auxin 3 indole butyric acid (AIB) and M3= M0+30g/l sugar + 0,5mg/l auxin AIB)\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-4200930/v1/85717aeb50974a8b321b922f.png"},{"id":54515241,"identity":"f912adaf-35f9-43f9-a3db-c7efb55799cd","added_by":"auto","created_at":"2024-04-11 16:24:20","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":74202,"visible":true,"origin":"","legend":"\u003cp\u003eCombined effect of sugar and AIB on in vitro plantlet growth parameters: a) stem height, b) number of roots, c) number of expanded leaves and d) in vitro plant weight. (Murashige and skoog 1962) 4,4g/l + 7g/l agar; M1= M0 + 30g/l sugar; M2= M0 + 0,5mg/l auxin 3 indole butyric acid (IBA) and M3= M0+30g/l sugar + 0,5mg/l auxin IBA)\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-4200930/v1/92d732f36ec04a104d657b54.png"},{"id":60650068,"identity":"9dc52a7a-3b56-4290-befd-364928ff9405","added_by":"auto","created_at":"2024-07-19 06:14:54","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":934186,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4200930/v1/89f144c9-f820-40c1-bcc6-f6cd220731fb.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Evaluation of four growth mediums effect on potato (Solanum tubérosum L.) plantlet production in Burkina Faso","fulltext":[{"header":"Introduction","content":"\u003cp\u003eSeeds are essential in most agricultural production systems and for farmers. Since the Green Revolution of the 1960s, the use of improved seeds have played an important role in the development of agriculture\u0026nbsp;[1]. Improved seed has been qualified as the engine of agricultural progress through its ability to increase productivity, improve incomes and food security\u0026nbsp;[1;2]. According to\u0026nbsp;[1]\u0026nbsp;seed technology is a set of applied sciences, technologies and socio-economic aspects that contribute to the production and availability of good quality seed for sowing. The potato seed is not excluded from this technology. Potato is considered the fourth most important food crop in the world after maize, rice and wheat\u0026nbsp;[3]. Globally, over the last 20 years, potato production has increased at a much higher rate in Asia, Africa and Latin America, rising from less than 30 million tonnes in the early 1960s to more than 165 million tonnes in 2007\u0026nbsp;[4]. Half of its current global production is in China and India\u0026nbsp;[4].The observed increase is thought to be due to the use of in vitro cultivation technology, which is used worldwide for seed production. According to\u0026nbsp;[5],\u0026nbsp;this technology has been used to produce disease-free seeds.\u0026nbsp;Potato seed can be produced via in vitro propagation in two ways, after micropropagation of the explants. The in vitro plantlets thus obtained can be used, after acclimatisation, to produce minitubers, or they can be induced to produce microtubers. The use of in vitro plantlets or microtubers for further seed production can also be achieved by several methods, such as hydroponics, aeroponics and many others.\u0026nbsp;According to\u0026nbsp;[6], aeroponics as promoted by the International Potato Center (CIP), has several advantages, including high multiplication rates.\u0026nbsp;This methodology is also used in sub-Saharan Africa including Burundi, Ethiopia, Kenya, Malawi, Rwanda, Tanzania and Uganda and has led to a large increase in minitubers production although this increase is still small\u0026nbsp;[7;8]. Despite this increase in potato production, the majority of developing countries, particularly those in West Africa, are faced difficulties in obtaining seed\u0026nbsp;[10;11]. Burkina Faso is not an exception to this constraint, potato growers are faced with difficulties in obtaining seed\u0026nbsp;[12]. According to statistics from the Ministry of Agriculture, Animal Resources and Fisheries, most of the seeds used by producers come from outside the country. In fact, two types of potato seed are used in Burkina Faso: farm- seed from previous production and certified seed imported from outside the country, especially from Europe. Imported seed, which are mainly used, only partially meets the required quality criteria and is not available in time\u0026nbsp;[7;12]. As a result, local seed production needs to be developed. In vitro culture technology through the micropropagation of vitroplants can help to alleviate this problem by providing growers with quality seeds. This technique requires the use of appropriate culture mediums. Various culture mediums composed of MS medium (Murashige and skoog 1962), sugar and auxin AIB have been tested. MS medium is generally used as a basic medium in many in vitro culture\u0026nbsp;[13;14;15]. A study by\u0026nbsp;[16]\u0026nbsp;showed the effectiveness of the MS medium (Murashige and skoog 1962) on the development of somatic embryos from the. picholine marocaine\u0026nbsp;(\u003cem\u003eOlea europaea L\u003c/em\u003e). Sucrose is the most effective source of carbohydrate [17]. According to [18], the use of table sugar has improved potato micropropagation but has also reduced production costs by 34 to 51% compared to analytical grade sucrose. Auxin is a growth hormone and is required for pollen embryo induction in many Solanaceae species except tobacco [19]. Hence our interest in using these elements. The purpose of our study was to identify a culture medium suitable for micropropagation of potato in vitro plantlets in Burkina Faso. Specifically, in combination with the MS base medium, we aimed to: i) evaluate the effect of sugar on the development of in vitro plantlets; ii) evaluate the effect of the auxin indole-3-butyric acid (IBA) on the development of in vitro plantlets; and iii) evaluate the combined effect of sugar and the auxin IBA on in vitro plantlets.\u0026nbsp;\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003e\u003cstrong\u003eMaterial\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe plant material is consisted of tubers of improved potato varieties and in vito plantlet genotypes from the International Potato Center (CIP). A total of two improved varieties (\u003cem\u003eSahel\u003c/em\u003e and \u003cem\u003eSpunta\u003c/em\u003e) and eight CIP genotypes were used for in vitro culture.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethodology\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe work was carried out at INERA station of Kamboins\u0026eacute;, in the in vitro culture laboratory. The tubers of the improved varieties underwent initiation, then multiplication before being used as plant material. The axillary buds of approximately 1 cm, consisting of a node of these in vitro plantlets, were used as explants and transplanted onto each of the four cultures mediums at a rate of 10 ml of culture medium per explant.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe culture medium consisted of \u003cstrong\u003e:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eM0= 4,4g/l MS (Murashige and skoog 1962) + 7g/l agar, M0 is the base medium;\u003c/p\u003e\n\u003cp\u003eM1= M0 + 30g/l sugar;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eM2= M0 + 0,5mg/l auxin 3 indole butyric acid (IBA) and\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eM3= M0+30g/l sugar + 0,5mg/l auxin IBA.\u003c/p\u003e\n\u003cp\u003eThe pH of each medium was adjusted to pH=5.7\u0026plusmn;0.1 with 1N NaOH. A 909 ml cylindrical glass jar was used as the culture vessel. The prepared media were dispensed into the containers and then autoclaved for 20 minutes at a pressure of 120 bar before subculturing in the jars under a laminar flow hood. These containers containing the explants were loosely covered and placed in a culture room for 30 days at a temperature of 22 \u0026plusmn; 1\u003csup\u003e\u0026nbsp;o\u003c/sup\u003eC, 70% relative humidity, a photoperiod of 16 h and a lighting intensity of 1200 lux.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eExcel 2016 was used for data entry.\u003c/p\u003e\n\u003cp\u003eXLSTAT 2016 was used to perform an analysis of variance of the data to determine the effect of the different environments on the growth parameters of the plantlets. The results of this analysis were used to produce tables, frequency calculations and histograms using Excel 2016.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eRegeneration rate of plantlets\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable1\u0026nbsp;\u003c/strong\u003eAnalysis of variance of environmental data\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Media\u003c/p\u003e\n \u003cp\u003eGrowth parameters\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eM0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eM1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eM2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eM3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003ePr \u0026gt; F\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eNRE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,092 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,817 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,067 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,767 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n 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width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e1,675 c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eLFE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,002 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,025 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,044 c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eNNO\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,158 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e5,000 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,067 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e4,846 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eDEN\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,010 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,371 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,529 c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eNRM\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,321 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,571 c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eNOR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,017 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e1,783 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,117 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e4,321 c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eLRA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,002 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,371 c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,001 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,304 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003ePPD\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,001 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,016 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,001 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,026 c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003eNRA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,025 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,254 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000 a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,233 b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.666666666666668%\" valign=\"top\"\u003e\n \u003cp\u003e0,000\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003eNRE: Number of shoots; HTI: Height of stem (cm), DTI: Diameter of stem (mm), NFE: Number of leaves, LFE: Length of leaf (cm), NNO: Number of nodes, DEN: Distance between nodes (cm), NRM: Number of branches, NOR: Number of roots, LRA: Length of roots (cm), PPD: Weight of in vitro plants (g), NRA: Number of aerial roots.\u003c/p\u003e\n\u003cp\u003eThe result of the analysis of variance showed a very highly significant difference between the growth parameters of the different media with a Pr \u0026gt; F (0,0001) (Table 1). Statistical analysis also showed that all the media used favoured vegetative regeneration of in vitro plantlets, with the rate varying from one medium to another. As a result, mediums M1 and M3 showed the best regeneration rates, ranging from 46,89% to 44,02% respectively. However, low regeneration rates were observed in media M0 and M2, at 5,26% and 3,83% respectively (\u003cstrong\u003eFig. 1\u003c/strong\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026nbsp;Effect of sugar on the development of in vitro plantlets\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe results showed that sugar had a positive effect on 12 growth parameters measured. The parameters most affected were: root length, stem diameter, number of nodes and number of leaves open. Root length varied from 0,05 to 2,2cm. Long roots of more than 0,5 cm were observed in medium M1, with a frequency of around 63,87% of in vitro plants. The diameter of the plantlets varied from 0,5 to 4 mm. The majority of plantlets (81,04%) had a diameter of between 0,05 and 0.1 cm and were observed in the M1 medium. The number of nodes and the number of opened leaves varied from 1 to 14 nodes and from 1 to 7 leaves, respectively. The majority of plantlets with a high node number of more than 5 and a number of opened leaves of between 1 and 4 were observed in medium M1, with a frequency of 72,21% and 79,75% of plantlets respectively (\u003cstrong\u003eFig\u003c/strong\u003e \u003cstrong\u003e2\u003c/strong\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEffect of auxin indole-3-butyric acid (IBA)\u003c/strong\u003e \u003cstrong\u003eon the development of in vitro plantlets\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis analysis showed that auxin indole-3-butyric acid (IBA) had a positive effect on root proliferation and calogenesis. In fact, we observed root proliferation in medium M2 with 5,83% of in vitro plantlets (Fig.3).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCombined effect of sugar and auxin indole-3-butyric acid (IBA) on the development of in vitro plantlets\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis analysis shows that the combined effect of sugar and auxin AIB was positive on in vitro plant growth parameters. The parameters most affected were: stem height, number of roots, number of leaves open and weight of in vitro plantlets, which strongly differentiated M3 from the other media. The height of the plantlets varied from 0,1 to 8 cm. In fact, 29,67% of in vitro plantlets with a height of between 4 and 6 cm were observed in medium M3. The number of roots varied from 1 to 15, with 46,22% of the plantlets in medium M3 having between 6 and 15 roots. The number of opened leaves varied from 1 to 12 leaves, 46,61% of the plantlets in medium M3 had an opened leaves number between 1 and 4. The weight of the plantlets varied from 0,003 to 0,1g, 41,34% of the M3 plantlets had a weight of between 0,003 and 0,05g (Fig. 4).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe low regeneration of in vitro plantlets in mediums M0 and M2 could be due to a lack of sugar in both mediums. Sugar would be the missing element in these media compared to mediums M1 and M3, which showed good regeneration (more than 5%). In this sense\u0026nbsp;[20]\u0026nbsp;showed that sugar is an essential element for both the vegetative development of cuttings and microtuberisation. According to these authors, the presence of sugar in a growing medium is favorable to cauline and root growth. Our results showed a positive effect of sugar in the M1 medium on growth parameters. The number of nodes, number of leaves open, stem diameter and root length were the parameters most influenced by the effect of sugar. The 30 g/l sugar dose of the M1 medium produced in vitro plantlets with more than 5 nodes and a number of open leaves of between 1 and 4, with a stem diameter of around 1mm and a root length of more than 0,5 cm. We can therefore say that sugar at this dose favours the stem and root development of in vitro plantlets. According to\u0026nbsp;[18], table sugar improves potato micropropagation. Sugar is therefore a source of energy used by explants for their growth and development. The same applies to exogenous sucrose, which is thought to be directly involved in regulating cell proliferation, photosynthesis and the defence mechanism against reactive oxygen derivatives\u0026nbsp;[13]. According to\u0026nbsp;[21]\u0026nbsp;sucrose is the best source of carbon in \u003cem\u003eIpomoea batatas L\u003c/em\u003e, followed by fructose and glucose. Furthermore,\u0026nbsp;[14], have shown that sucrose, glucose and maltose have a favourable effect on various growth parameters in potatoes. We can therefore say that in vitro multiplication of vegetative explants would require a carbon source such as sugar.\u003c/p\u003e\n\u003cp\u003eThe root development of in vitro plantlets in the M2 medium is thought to be due to the effect of auxin AIB. Auxin is thought to be produced in the apical buds of plants and transported from stems to roots via a polar transport system\u0026nbsp;[22]. It is accumulated in the root tips and stimulates rhizogenesis by favouring the initiation and development of lateral roots\u0026nbsp;[22]. According to\u0026nbsp;[23;24], AIB positively improves the rooting phase in in vitro culture in cork oak and apple.\u0026nbsp;[21]\u0026nbsp;showed that auxin AIB gave the best rooting parameters compared to ANA for root formation in \u003cem\u003eIpomoea batatas L\u003c/em\u003e. Auxin is known to regulate many aspects of plant growth and development, such as vascular tissue differentiation, embryonic development, root formation and primary and secondary stem formation\u0026nbsp;[25]. Explant regeneration was promoted by the effect of sugar. This regeneration is essential for the production and assimilation of auxin in vitro plantlets. As a result, the combination of sugar and auxin will accentuate the action of auxin through high root production and will allow good development of the in vitro plantlets observed in the M3 medium compared to the other mediums. Indeed,\u0026nbsp;[26;27]\u0026nbsp;reported that the application of glucose, sucrose and fructose stimulated the development of seedlings of \u0026nbsp;\u003cem\u003eDactylorhiza species\u003c/em\u003e and \u003cem\u003ePhysocarpus opulifolius\u003c/em\u003e (L.) and that the growth rate and root length of seedlings increased in the presence of AIB and\u0026nbsp;\u0026alpha;-naphthalene-acetic acid. Thus, the combined effect of sugar and auxin AIB allows the regeneration of explants and the growth of their roots. The in vitro plantlets in medium M3 were characterised by an average height, a higher number of roots, a higher number of leaves and a higher weight of plantlets compared to those in medium. These characteristics would be very important for good acclimatisation of the plantlets. Indeed\u0026nbsp;[21]\u0026nbsp;showed that the better rooting rate observed with AIB favoured 100% success at the acclimatisation stage in sweet potato (\u003cem\u003eIpomoea batatas L)\u003c/em\u003e.\u0026nbsp;\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study revealed that the four media used favour the regeneration of in vitro plantlets, but M1 and M3 are the best mediums for good vegetative regeneration. Sugar had a positive effect on growth parameters. Sugar favoured the development of stem diameter, root length, number of nodes and fuller leaves. AIB auxin had a positive effect on root proliferation of in vitro plantlets. The combination of sugar and auxin AIB improved the development of growth parameters such as stem height, number of roots, number of expanded leaves and the weight of in vitro plantlets in medium M3 compared with the other mediums. Thus, the production of plantlets showing good vegetative growth would allow good acclimatisation an improvement of the production of potato seeds.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAvailability of data\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data used to support the results of this study will be available on request from the corresponding author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that there are no conflicts of interest in the publication of this article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the members of the Plant Genetics and Improvement team of the Biosciences Laboratory of University Joseph KI-ZERBO and the Institute of the Environment and Agricultural Research (INERA) Kamboins\u0026eacute;, more specifically the members of the in vitro culture laboratory, for their collaboration in carrying out this study. They would also like to thank Dr Idrissa OUEDRAOGO of the Centre Universitaire de Dori / Universit\u0026eacute; Thomas SANKARA for his advice and suggestions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics declarations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA licence has been obtained for the use of the plant material. The study complies with national guidelines.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eM. Turner, Les semences Quae Cta Presses agronomiques de Gembloux. 2010.\u003c/li\u003e\n\u003cli\u003eM. Cohen et al., Cartes sur table. .\u003c/li\u003e\n\u003cli\u003eW. Food, World Food and Agriculture \u0026ndash; Statistical Yearbook 2022. 2022.\u003c/li\u003e\n\u003cli\u003eA. Devaux, P. Kromann, and O. Ortiz, \u0026ldquo;Potatoes for Sustainable Global Food Security,\u0026rdquo; Potato Res., vol. 57, no. 3\u0026ndash;4, pp. 185\u0026ndash;199, 2014, doi: 10.1007/s11540-014-9265-1.\u003c/li\u003e\n\u003cli\u003eP. Wang and C. 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Ortiz, The potato crop: Its agricultural, nutritional and social contribution to humankind. Springer International Publishing, 2019.\u003c/li\u003e\n\u003cli\u003eD. Harahagazwe et al., \u0026ldquo;How big is the potato (Solanum tuberosum L.) yield gap in Sub-Saharan Africa and why? A participatory approach,\u0026rdquo; Open Agric., vol. 3, no. 1, pp. 180\u0026ndash;189, Jan. 2018, doi: 10.1515/opag-2018-0019.\u003c/li\u003e\n\u003cli\u003eAmina. Belguendouz, \u0026ldquo;Essai de substitution des milieux de culture en micropropagation et la physiologie de la microtub\u0026eacute;risation de la pomme de terre (Solanum tuberosum. L),\u0026rdquo; pp. 1\u0026ndash;184, 2011.\u003c/li\u003e\n\u003cli\u003eM. Abdoulaye, \u0026ldquo;Production of potato (Solanum tuberosum L) pre-basic seed through tissue culture in katibougou, MALI,\u0026rdquo; University of Ghana, 2018.\u003c/li\u003e\n\u003cli\u003eA. Zerbo et al., \u0026ldquo;Potato ( Solanum tuberosum L . ) in Burkina Faso : cultivated varieties and production constraints,\u0026rdquo; pp. 18633\u0026ndash;18643, 2022.\u003c/li\u003e\n\u003cli\u003eJ. . Dubuc, \u0026ldquo;Iccumulationmpact des conditions de culturein vitro et du saccharoseexog\u0026egrave;nesur la r\u0026eacute;gulationde l\u0026rsquo;expression g\u0026eacute;niqueet l\u0026rsquo;accumulation des prot\u0026eacute;ine chez les plantules de tomate ( Solanum lycopersicum),\u0026rdquo; Universit\u0026eacute; Laval, 2010.\u003c/li\u003e\n\u003cli\u003eM. H. Rahman, R. Islam, M. Hossain, and M. S. Islam, \u0026ldquo;Role of sucrose , glucose and maltose on conventional potato micropropagation,\u0026rdquo; J. Agric. Technol., vol. 6, no. 4, pp. 733\u0026ndash;739, 2010.\u003c/li\u003e\n\u003cli\u003eL. Chen, Y. Lu, Y. Hu, and X. 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Ewelina, \u0026ldquo;The effect of carbon source in culture medium on micropropagation of common ninebark ( Physocarpus opulifolius ( L .) Maxim .) \u0026lsquo; Diable D \u0026rsquo; or \u0026rsquo; Agnieszka Ilczuk , Katarzyna Jagie łł o-Kubiec , Ewelina Jacygrad,\u0026rdquo; vol. 12, no. 3, pp. 23\u0026ndash;33, 2013.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"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":"Micropropagation, sugar, auxin AIB, in vitro plantlets, potato and Burkina Faso","lastPublishedDoi":"10.21203/rs.3.rs-4200930/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4200930/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"In Burkina Faso, potato growers are faced with difficulties in obtaining seed. These seeds, mostly imported partially meet the quality requirement in potato production. Furthermore, these seeds are not accessible in term of cost and also not available in time. Local seed production is therefore necessary. In vitro culture can remedy this problem by providing quality seeds. Thus, the goal of our study was to set up an in vitro culture protocol for the production of plantlets with optimal vegetative growth. This study was realised in the in vitro culture laboratory at INERA Kamboinsé. Explants from in vitro plantlets (1 cm) of the same generation were used as plant material and transplanted onto four different mediums. The mediums used were: M0= 4,4g/l MS medium (Murashige and Skoog 1962) + 7g/l agar; M1= M0 + 30g/l table sugar; M2= M0 + 0,5mg/l AIB and M3= M0+30g/l sugar + 0,5mg/l AIB. These mediums were prepared with a pH=5,7±0.1, then divided into jars and autoclaved for 20 minutes at a pressure of 120 bars. Subculture in the jars was realised out in a laminar flow hood. The covered jars were transferred to a growth room for 30 days. The results of this study indicated that M1 and M3 showed good regeneration of the in vitro plantlets compared to M0 and M2. Sugar probably had a positive effect on root length, stem diameter, number of nodes and number of leaves open. These parameters strongly differentiated M1 from the other mediums. AIB had a positive effect on the root proliferation of in vitro plantlets in the M2 medium. The combined effect of sugar and AIB was even more positive on stem height, number of roots, number of leaves open and weight of in vitro plantlets, which strongly differentiated medium M3 from the other mediums. In conclusion, M3 proved to be the best medium for producing in vitro plantlets with good vegetative growth for seed production.","manuscriptTitle":"Evaluation of four growth mediums effect on potato (Solanum tubérosum L.) plantlet production in Burkina Faso","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-11 16:24:14","doi":"10.21203/rs.3.rs-4200930/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":"309080e7-3c41-4201-b9fe-b1ddb34295b3","owner":[],"postedDate":"April 11th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-07-19T06:06:47+00:00","versionOfRecord":[],"versionCreatedAt":"2024-04-11 16:24:14","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4200930","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4200930","identity":"rs-4200930","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}