Effects of Different Hormonal Concentrations on In vitro Regeneration and Multiplication of Pearl Millet (Pennisetum glaucum L.)

R. Khan, Robina Ilyas, Saqib Saleem, Shahnaz Bibi, M. R. Khan, M. Younas, Matee ur Rehman Hafiz, S. Ullah, Basharat Mahmood
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Abstract

An efficient in-vitro regeneration and multiplication protocol was developed to check the effects of different hormonal concentrations through somatic embryogenesis of Pearl millet (Pennisetum glaucum L.). The explants (seeds) of P. glaucum were surface sterilized with different concentrations (30, 50 and 70%) of Sodium hypochlorite to ensure the removal of surface contamination. For the regeneration the explants were inoculated on MS media supplemented with varying concentrations of indole acetic acid (IAA 2 mg/L, 0.75 mg/L, 0.1 mg/L, 0.2 mg/L and 0.3 mg/L) and kinetin (KIN 1.5 mg/L, 0.5 mg/L, 0.5 mg/L, 1 mg/L and 1.5mg/L) respectively. The maximum germination (87.5 %) of explants was obtained by using 70 % Chlorox and minimum germination (62.5 %) of explants was obtained by using 30% Chlorox. The maximum stem length (15.5 cm), roots number (12), roots length (3.96 cm) and leaves length (6.76 cm) was observed on MS medium containing (IAA 2 mg/L and KIN 1.5 mg/L). The maximum leaves number (7) was observed on MS medium containing (IAA 0.75 mg/L and KIN 0.5 mg/L). The minimum stem length (3 cm), roots number (1), roots length (1 cm), leaves number (1) and leaves length (3 cm) was observed on MS medium supplemented with (IAA 0.2 mg/L and KIN 1 mg/L). The maximum Stem length (15.5cm), Roots number (12), Roots length (3.96 cm)  and Leaves length (6.76 cm) was observed on MS medium supplemented with (IAA 2 mg/L and KIN 1.5 mg/L) although maximum leaves number was observed on MS medium supplemented with (IAA 0.75 mg/L and KIN 0.5 mg/L). The study revealed an easy and reproducible in-vitro regeneration protocol of pearl millet that can provide an efficient plant regeneration which can be further exploited for transgenic applications
不同激素浓度对珍珠粟(Pennisetum glaucum L.)体外再生和增殖的影响
为研究不同激素浓度对珍珠粟体细胞胚发生的影响,建立了一种高效的体外再生增殖方案。采用不同浓度的次氯酸钠(30%、50%和70%)对青稞外植体(种子)进行表面消毒,以确保表面污染物的去除。将外植体接种于分别添加不同浓度吲哚乙酸(IAA 2 mg/L、0.75 mg/L、0.1 mg/L、0.2 mg/L和0.3 mg/L)和kinetin (KIN 1.5mg/L、0.5 mg/L、0.5 mg/L、1 mg/L和1.5mg/L)的MS培养基上进行再生。70% Chlorox处理的外植体萌发率最高(87.5%),30% Chlorox处理的外植体萌发率最低(62.5%)。在IAA含量为2 mg/L、KIN含量为1.5 mg/L的MS培养基上,植株茎长最大(15.5 cm),根数最大(12根),根长最大(3.96 cm),叶长最大(6.76 cm)。在含IAA 0.75 mg/L、KIN 0.5 mg/L的MS培养基上,叶片数最多,为7个;在添加(IAA 0.2 mg/L和KIN 1 mg/L)的MS培养基上观察最小茎长(3 cm)、根数(1)、根长(1 cm)、叶数(1)和叶长(3 cm)。在添加(IAA 2 mg/L和KIN 1.5 mg/L)的MS培养基上,茎长(15.5cm)、根数(12)、根长(3.96 cm)和叶长(6.76 cm)最大,而在添加(IAA 0.75 mg/L和KIN 0.5 mg/L)的MS培养基上,叶数最大。本研究揭示了一种简单、可重复的珍珠粟离体再生方案,为珍珠粟的转基因应用提供了有效的植株再生途径
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