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Transcriptome Analysis of a New Peanut Seed Coat Mutant for the Physiological Regulatory Mechanism Involved in Seed Coat Cracking and Pigmentation

机译:一种新的花生种皮突变体的转录组分析涉及种子皮破裂和色素沉着的生理调控机制。

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摘要

Seed-coat cracking and undesirable color of seed coat highly affects external appearance and commercial value of peanuts (Arachis hypogaea L.). With an objective to find genetic solution to the above problems, a peanut mutant with cracking and brown colored seed coat (testa) was identified from an EMS treated mutant population and designated as “peanut seed coat crack and brown color mutant line (pscb).” The seed coat weight of the mutant was almost twice of the wild type, and the germination time was significantly shorter than wild type. Further, the mutant had lower level of lignin, anthocyanin, proanthocyanidin content, and highly increased level of melanin content as compared to wild type. Using RNA-Seq, we examined the seed coat transcriptome in three stages of seed development in the wild type and the pscb mutant. The RNA-Seq analysis revealed presence of highly differentially expressed phenylpropanoid and flavonoid pathway genes in all the three seed development stages, especially at 40 days after flowering (DAF40). Also, the expression of polyphenol oxidases and peroxidase were found to be activated significantly especially in the late seed developmental stage. The genome-wide comparative study of the expression profiles revealed 62 differentially expressed genes common across all the three stages. By analyzing the expression patterns and the sequences of the common differentially expressed genes of the three stages, three candidate genes namely c36498_g1 (CCoAOMT1), c40902_g2 (kinesin), and c33560_g1 (MYB3) were identified responsible for seed-coat cracking and brown color phenotype. Therefore, this study not only provided candidate genes but also provided greater insights and molecular genetic control of peanut seed-coat cracking and color variation. The information generated in this study will facilitate further identification of causal gene and diagnostic markers for breeding improved peanut varieties with smooth and desirable seed coat color.
机译:种皮开裂和种皮的不良颜色极大地影响了花生的外观和商业价值(花生)。为了找到解决上述问题的遗传方法,从经EMS处理的突变群体中鉴定出具有裂纹和褐色种皮(种皮)的花生突变体,并命名为“花生种皮裂纹和褐色突变体系(pscb)”。 ”突变体的种皮重量几乎是野生型的两倍,并且发芽时间明显短于野生型。此外,与野生型相比,该突变体具有较低水平的木质素,花色苷,原花色素含量,并具有高度增加的黑色素含量水平。使用RNA-Seq,我们在野生型和pscb突变体的种子发育的三个阶段检查了种皮转录组。 RNA-Seq分析显示,在所有三个种子发育阶段,特别是在开花后40天(DAF40),都存在高度差异表达的苯丙烷和类黄酮途径基因。另外,发现多酚氧化酶和过氧化物酶的表达被显着激活,尤其是在种子发育后期。表达谱的全基因组比较研究揭示了在所有三个阶段共有的62个差异表达基因。通过分析这三个阶段的共同差异表达基因的表达模式和序列,确定了三个候选基因,分别为c36498_g1(CCoAOMT1),c40902_g2(驱动蛋白)和c33560_g1(MYB3),其负责种皮破裂和褐色表型。 。因此,这项研究不仅提供了候选基因,而且为花生种皮的破裂和颜色变化提供了更深刻的见解和分子遗传控制。本研究中产生的信息将有助于进一步鉴定因果基因和诊断标记,以育种具有光滑且理想的种皮颜色的改良花生品种。

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