Cassava is a staple food for over 800 million people globally providing a cheap source of carbohydrate. However, the cultivation of cassava in the country is facing to viral diseases, particularly cassava mosaic disease (CMD) which can cause up to 95% yield losses. With aim to supply farmers demand for clean planting materials, there is need to accelerate the production of the elite cultivars by use of tissue culture in order to cope with the demand. Nodal explants harvested from the greenhouse grown plants were sterilised using different concentrations of a commercial bleach JIK (3.85% NaOCl) and varying time intervals. Microshoots induction was evaluated using thidiazuron (TDZ), benzyl amino purine (BAP), and kinetin. Rooting was evaluated using different auxins (Naphthalene acetic acid NAA and Indole-3-butyricacid IBA). PCR-based SSR and SCAR markers were used to verify the presence of CMD2 gene in the regenerated plantlets. The highest level of sterility in explants (90%) was obtained when 20% Jik was used for 15 min. The best cytokinin for microshoots regeneration was found to be kinetin with optimum concentrations of 5, 10 and 20 μM for Agric-rouge, Atinwewe, and Agblehoundo respectively. Medium without growth regulators was the best for rooting the three cultivars. A survival rate of 100, 98, and 98% was recorded in the greenhouse for Agric-rouge, Atinwewe, and Agblehoundo respectively and the plantlets appeared to be morphologically normal. The SSR and SCAR analysis of micropropagated plants showed a profile similar to that of the mother plants indicating that the regenerated plantlets retained the CMD2 gene after passing through in vitro culture, as expected with micropropagation. The nodal explants was established to be 20% of Jik (3.85% NaOCl) with an exposure time of 15 min. Kinetin was proved to be the best cytokinins for microshoot formation with the optimum concentration of 5, 10 and 20 μM for Agric-rouge, Atinwewe, and Agblehoundo respectively. The protocol developed during this study will be useful for mass propagation of the elite cassava cultivars.

中文翻译：

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三个抗花叶病木薯品种的离体繁殖

木薯是全球超过 8 亿人的主食，提供廉价的碳水化合物来源。然而，该国的木薯种植正面临病毒性疾病，特别是木薯花叶病（CMD），可导致高达 95% 的产量损失。为了满足农民对清洁种植材料的需求，需要通过组织培养加速优良品种的生产以满足需求。使用不同浓度的商业漂白剂 JIK (3.85% NaOCl) 和不同的时间间隔对从温室种植的植物中收获的节点外植体进行消毒。使用噻二唑隆 (TDZ)、苄基氨基嘌呤 (BAP) 和激动素评估微芽诱导。使用不同的生长素（萘乙酸 NAA 和 Indole-3-butyricacid IBA）评估生根。使用基于 PCR 的 SSR 和 SCAR 标记来验证再生植株中 CMD2 基因的存在。当使用 20% Jik 15 分钟时，外植体的无菌水平最高 (90%)。发现用于微芽再生的最佳细胞分裂素是激动素，对于 Agric-rouge、Atinwewe 和 Agblehoundo，最佳浓度分别为 5、10 和 20 μM。不含生长调节剂的培养基最适合三个品种的生根。在温室中记录的 Agric-rouge、Atinwewe 和 Agblehoundo 的成活率分别为 100、98 和 98%，并且幼苗的形态正常。微繁植株的 SSR 和 SCAR 分析显示出与母株相似的特征，表明再生植株经过体外培养后保留了 CMD2 基因，正如微繁殖所预期的那样。节点外植体被确定为 20% 的 Jik（3.85% NaOCl），暴露时间为 15 分钟。激动素被证明是最好的微芽形成细胞分裂素，对于 Agric-rouge、Atinwewe 和 Agblehoundo 的最佳浓度分别为 5、10 和 20 μM。本研究期间制定的协议将有助于优良木薯品种的大规模繁殖。