Tewelde Selam, Patharajan Subban, Teka Zenebe, Sbhatu Desta Berhe
Tigrai Biotechnology Center Pvt. Ltd. Co., Mekelle, Ethiopia.
Aksum University, P.O. Box 1010, Aksum, Ethiopia.
ScientificWorldJournal. 2020 Jun 13;2020:6431301. doi: 10.1155/2020/6431301. eCollection 2020.
Ginger ( Rosc) (Zingiberaceae) is a livelihood and commercial crop in Ethiopia. But, the availability of clean and healthy planting materials has become a problem due to wilt disease, caused by Biovar 3 Race 4. This problem obliged growers to seek for tens of millions of vigorous and disease-free planting materials very quickly via micropropagation of shoot tip explants. For this purpose, protocols of sterilizing shoot tip explants and controlling bacterial contamination of one Ethiopian ginger cultivar called Deribo were tested. Hence, this article reports the finding of a study that aimed at testing the (a) effectiveness of three sterilization agents, namely, 0.25% w/v RBK (composed of ridomile, bayleton, and kocide at 1 : 1 : 1 ratio), 0.50% v/v NaOCl, and 70% v/v ethanol at three different treatment times in combination with 0.25% HgCl; (b) efficacy of four broad-spectrum antibiotics and their combinations in controlling bacterial contaminants of ginger shoot tip explants and micropropagation media; and (c) effects of the antibiotics on the shooting performances of the explants of the cultivar. A 0.50% v/v NaOCl at exposure time of 20 min followed by 0.25% HgCl has resulted in 80% contamination-free and 70% live explants after three weeks of incubation. Likewise, cefotaxime at 50, 100, and 200 mg/L and cefotaxime plus streptomycin at 25, 50, and 100 mg/L yielded 87 to 93% contamination-free microshoots after three weeks of culturing. The number of explants killed by the antibiotics increased with increasing the concentration of the antibiotics. Cefotaxime at 50 mg/L and cefotaxime plus streptomycin at 25 mg/L yielded significantly highest mean microshoots per explant (7.10 ± 0.36 and 7.51 ± 0.27, respectively) and mean shoot length (4.2 ± 0.26 and 3.56 ± 0.17 cm, respectively). Some of the microshoots showed some yellowing. But, they turned green and grew normal after subcultured into fresh, antibiotics-free culture media. These findings are important foundations towards developing more optimized protocols of sterilizing explants and controlling bacterial contaminants for large-scale micropropagation of the Deribo ginger cultivar.
姜(Roscoe)(姜科)是埃塞俄比亚的一种民生和经济作物。但是,由于由生物变种3小种4引起的枯萎病,清洁健康的种植材料的供应已成为一个问题。这个问题迫使种植者通过茎尖外植体的微繁殖迅速寻求数千万株健壮且无病的种植材料。为此,对一种名为德里博的埃塞俄比亚姜品种的茎尖外植体消毒和控制细菌污染的方案进行了测试。因此,本文报告了一项研究的结果,该研究旨在测试:(a)三种消毒剂,即0.25%(w/v)RBK(由甲霜灵、百菌清和代森锰锌按1∶1∶1比例组成)、0.50%(v/v)次氯酸钠和70%(v/v)乙醇在三个不同处理时间与0.25%氯化汞组合使用的有效性;(b)四种广谱抗生素及其组合在控制姜茎尖外植体和微繁殖培养基细菌污染物方面的功效;(c)抗生素对该品种外植体出芽性能的影响。在20分钟暴露时间下使用0.50%(v/v)次氯酸钠,然后使用0.25%氯化汞,培养三周后得到80%无污染和70%存活的外植体。同样,50、100和200mg/L的头孢噻肟以及25、50和100mg/L的头孢噻肟加链霉素在培养三周后产生了87%至93%无污染的微芽。被抗生素杀死的外植体数量随着抗生素浓度的增加而增加。50mg/L的头孢噻肟和浓度为25mg/L的头孢噻肟加链霉素分别产生了每个外植体显著最高的平均微芽数(分别为7.10±0.36和7.51±0.27)和平均芽长(分别为4.2±0.26和3.56±0.17cm)。一些微芽出现了一些发黄现象。但是,将它们转接至新鲜的、不含抗生素的培养基中后,它们变绿并正常生长。这些发现是开发更优化的外植体消毒和控制细菌污染物方案以用于德里博姜品种大规模微繁殖的重要基础。