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首页> 外文期刊>African Journal of Biotechnology >The study of 2,4-D and 2,4,5-T effects on gene expression at early stages of embryogenesis in chickpea (Cicer arientinum L.)
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The study of 2,4-D and 2,4,5-T effects on gene expression at early stages of embryogenesis in chickpea (Cicer arientinum L.)

机译：2,4-D和2,4,5-T对鹰嘴豆胚胎发生早期基因表达的影响研究

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Factors affecting the induction of embryogenesis in somatic plant cells?in vitro?are highly variable depending on different growth regulators, explants types, culture media and stress treatments. A series of experiments were set to study the effect of some factors involve in the initiation and development of somatic embryos including genotype, growth regulator and the kind of explants in four Kabuli type chickpea genotypes (F3, Jam, Bivanig and ILC485). Mature embryos were transferred on Murashige and Skooges (MS) medium plus 3% sucrose supplemented with two kind of auxin as growth regulators in order to induce embryogenesis. The best induction media (found in the preliminary experiment) was the one supplemented with either 3 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) or 3 mg/l 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) in single use or in combination, to induce?globular embryos. Globular shape embryos were distinguishable 6 to 7 weeks after culture.?Desiccation treatments were applied by keeping the samples for 7, 14 and 20 min in laminar air flow to induce globular embryos maturity.?Total RNAs were isolated using guanidium isothiocyanate method. The polymerase chain reaction amplification (PCR) reaction was performed using three one-base-anchored oligo-dT primers, in combination with eight arbitrary primers. Two genotypes of Bivanig and ILC482 were selected as explants for gene expression studies due to their highest number of globular embryos production, then 3 mg/l 2,4,5-T and 3 mg/l 2,4-D were used to produce embryogenic and non-embryogenic calli, respectively. The results showed a similar gene expression pattern in both treatments with three eminent steps.?Comparison of both similar band patterns of two auxin treatments indicated that, a number of particular genes are involved in cells determination. Different growth regulators and thegenotypes?created little changes in gene expression pattern through theembryogenesis process just before?the globular stage, resulting that, a large number of changes and genes occur similarly in both genotypes and auxin treatments.
机译：取决于不同的生长调节剂，外植体类型，培养基和胁迫处理，影响体细胞体外诱导胚胎发生的因素是高度可变的。进行了一系列实验来研究影响体细胞胚萌发和发育的一些因素的影响，包括基因型，生长调节剂和四种卡布利型鹰嘴豆基因型（F3，Jam，Bivanig和ILC485）的外植体种类。将成熟的胚胎转移到Murashige和Skooges（MS）培养基中，在培养基中添加3％的蔗糖，辅以两种生长素作为生长调节剂，以诱导胚胎发生。最好的诱导培养基（在初步实验中发现）是补充有3 mg / l的2,4-二氯苯氧基乙酸（2,4-D）或3 mg / l的2,4,5-三氯苯氧基乙酸（2， 4,5-T）单独使用或组合使用，以诱导球状胚胎。培养后6至7周可辨认出球形胚。？将样品在层流中保持7、14和20分钟进行干燥处理，以诱导球形胚成熟。？用异硫氰酸胍分离总RNA。聚合酶链反应扩增（PCR）反应是使用三个一碱基锚定的oligo-dT引物，与八个任意引物组合进行的。由于Bivanig和ILC482这两种基因型的球状胚产生数量最多，因此被选作外植体进行基因表达研究，然后使用3 mg / l 2,4,5-T和3 mg / l 2,4-D进行外源表达胚性和非胚性愈伤组织分别。结果表明，在两个处理中，三个显着步骤均具有相似的基因表达模式。比较两种生长素处理的两个相似谱带模式表明，许多特定基因参与细胞测定。不同的生长调节剂和基因型在球形阶段之前的胚发生过程中，基因表达模式几乎没有变化，从而导致大量的变化和基因在基因型和生长素处理中相似地发生。

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