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首页> 外文期刊>In Vitro Cellular and Development Biology. Plant: Journal of the Tissue Culture Association >Responses of Arabica coffee (Coffea arabica L. var. Catuai) cell suspensions to chemically induced mutagenesis and salinity stress under in vitro culture conditions
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Responses of Arabica coffee (Coffea arabica L. var. Catuai) cell suspensions to chemically induced mutagenesis and salinity stress under in vitro culture conditions

机译：阿拉伯咖啡（Coffea Arabica L.Var。Catuai）细胞悬浮液在体外培养条件下的化学诱导诱变和盐度应力

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Crop improvement of Coffea arabica L. (coffee) via mutagenesis could accelerate breeding programs; thus, the present study aimed to develop an in vitro protocol using the chemical mutagens sodium azide (NaN3) and ethyl methanesulfonate (EMS) on embryogenic cell suspensions of Arabica coffee variety Catuai and, subsequently, to evaluate the responses of the resulting mutagenized tissues to salinity stress. Embryogenic suspension cultures were incubated with 0.0, 2.5, 5.0, or 10.0 mM NaN3 or 0.0, 185.2, 370.5, or 741.0 mM EMS. As the concentration of NaN3 or EMS increased, the survival of embryogenic suspension cultures decreased compared to controls. The median lethal dose (LD50) for NaN3 was 5 mM for 15 min and for EMS it was 185.2 mM for 120 min. Embryogenic suspension cultures treated with NaN3 or EMS were cultured on selective medium supplemented with 0, 50, 100, 150, 250, or 300 mM NaCl showed that 50 mM NaCl could be used as selection pressure. Plantlet growth and total amino acid content were affected by NaCl stress; some mutants had longer shoots and higher amino acid content than controls. Random amplified polymorphic DNA (RAPD) analysis was performed to determine whether the NaN3 or EMS treatments could induce genetic variability and resulted in identifiable polymorphic markers. A total of 18 10-mer primers were used to amplify genomic DNA of putative mutant and non-mutant arabica coffee embryogenic cultures and produced 50 scorable bands, of which 22% were polymorphic.
机译：咖啡阿拉比卡咖啡酵母的作物改善（咖啡）通过诱变可以加速繁殖计划;因此，本研究旨在使用化学诱变叠氮化钠（NAN3）和甲磺酸乙酯（EMS）在阿拉伯咖啡品种CATUAI的胚性细胞悬浮液上进行体外方案，以及随后评估所得诱变组织的响应盐度压力。将胚胎悬浮液培养为0.0,2.5,5.0或10.0mM NaN 3或0.0,185.2,370.5或741.0mm EMS。随着NAN3或EMS的浓度增加，与对照相比，胚胎悬浮培养物的存活率降低。 NAN3的中值致命剂量（LD50）为5mm，15 mm，EMS为185.2mm，120分钟。在补充有0,50,100,150,250或300mM NaCl的选择性培养基上培养用NaN 3或EMS处理的胚胎悬浮液培养，显示50mM NaCl可用作选择压力。 Plantlet生长和总氨基酸含量受NaCl应激的影响;一些突变体具有比对照较长的芽和较高的氨基酸含量。进行随机扩增的多晶型DNA（RAPD）分析以确定NAN3或EMS治疗是否可以诱导遗传变异性并导致可识别的多态标记物。共18个10-MEL引物用于扩增推定突变体和非突变阿拉伯咖啡胚培养物的基因组DNA，并产生50个可批量的带，其中22％是多态的。

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