Zimik Masochon, Arumugam Neelakantan
Department of Biotechnology, School of Life Sciences, Pondicherry University, Kalapet, Pondicherry 605 014, India.
J Genet Eng Biotechnol. 2017 Dec;15(2):303-308. doi: 10.1016/j.jgeb.2017.07.006. Epub 2017 Jul 27.
is an ancient oilseed crop known for its high quality edible oil and its medicinally important lignans. The crop is said to be recalcitrant to plant tissue culture thus limiting the use of modern biotechnology for its genetic improvement. We present here a protocol describing plant regeneration through adventitious shoot formation from cotyledons dissected from sesame seeds soaked for four hours in water. Subculturing of the cotyledons after two weeks of culture on to a fresh Murashige and Skoog medium leads to differentiation of adventitious shoots from the proximal cut end of the explant. Culture of cotyledons on a medium containing 9% sucrose for a couple of weeks prior to transfer to MS medium supplemented with 3% sucrose induced a higher frequency of shoot regeneration. The highest frequency of 25% adventitious shoot regeneration was observed for variety UMA. This variety also turned out to be the best among the ten genotypes tested for shoot regeneration through tissue culture. While addition of IAA marginally improved regeneration, silver nitrate was found essential for enhancing the frequency of shoot regeneration. The regenerated shoots formed roots on full strength MS medium supplemented with 1 mg/l IBA and the rooted plants were established in soil.
是一种古老的油料作物，以其高品质的食用油和具有重要药用价值的木脂素而闻名。据说这种作物对植物组织培养具有顽拗性，因此限制了现代生物技术在其遗传改良中的应用。我们在此展示一种方案，描述从浸泡在水中4小时的芝麻种子中切取的子叶通过不定芽形成进行植株再生。在新鲜的Murashige和Skoog培养基上培养两周后，将子叶继代培养，可从外植体的近端切口端分化出不定芽。在转移到添加3%蔗糖的MS培养基之前，将子叶在含有9%蔗糖的培养基上培养几周，可诱导更高频率的芽再生。UMA品种观察到最高频率的25%不定芽再生。在通过组织培养进行芽再生测试的十个基因型中，该品种也是最好的。虽然添加吲哚乙酸对再生有轻微改善，但发现硝酸银对提高芽再生频率至关重要。再生芽在添加1毫克/升吲哚丁酸的全强度MS培养基上生根，生根的植株在土壤中定植。
