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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:新花生种子涂层突变体的转录组分析,涉及种子涂层裂纹和色素沉着的生理调节机制

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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:种子涂层开裂和种子涂层的不良颜色高度影响了花生的外观和商业价值(Arachis Hypogaea L.)。通过目的,寻找上述问题的遗传解决方案,从EMS处理的突变群体中鉴定了具有裂化和褐色种子涂层(Testa)的花生突变体,并指定为“花生种子涂层裂纹和棕色突变线(PSCB)。 “突变体的种子涂层重量几乎是野生型的两倍,萌发时间明显短于野生型。此外,与野生型相比,突变体具有较低水平的木质素,花青素,原花青素含量,并高度增加的黑色素含量水平。使用RNA-SEQ,我们在野生型和PSCB突变体中检查了种子发育的三个阶段的种子涂层转录组。 RNA-SEQ分析显示出在所有三种种子发育阶段的高差异表达苯基丙醇和黄酮类途径基因的存在,特别是在开花后40天(DAF40)。此外,发现多酚氧化酶和过氧化物酶的表达明显激活,特别是在晚期种子发育阶段。表达谱的基因组对比研究揭示了所有三个阶段常见的62个差异表达基因。通过分析三个阶段的常见差异表达基因的表达模式和序列,鉴定了三种候选基因即C36498_G1(CCOAOMT1),C40902_G2(Kinesin)和C33560_G1(MYB3),其负责种子涂层裂化和棕色表型。因此,这项研究不仅提供了候选基因,而且还提供了更大的洞察力和对花生种子涂层裂纹和颜色变异的分子遗传控制。本研究中产生的信息将有助于进一步鉴定因育种改善的花生品种而具有光滑和理想的种子涂料的造成因果基因和诊断标志物。

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