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应用 DNA 条码技术、SCoT 和 SDS-PAGE 蛋白分析微型繁殖无花果(L.)植株的体细胞无性系变异。

Using of DNA-Barcoding, SCoT and SDS-PAGE Protein to Assess Soma-Clonal Variation in Micro-Propagated Fig ( L.) Plant.

出版信息

Pak J Biol Sci. 2022 Jan;25(5):415-425. doi: 10.3923/pjbs.2022.415.425.

Abstract

<b>Background and Objective:</b> <i>In vitro</i> propagation of fig (<i>Ficus carica</i> L.) is one of the possible approaches that may be used to maximize the diversity of plant species. The current work was carried out to evaluate genetic stability of micropropagated fig plantlets and to determine the effect of <i>in vitro </i>propagation on genomic content of Saudi fig. <b>Materials and Methods:</b> The start codon-targeted (SCoT), DNA-barcoding chloroplast gene RNA polymerase1 (<i>rpoC1</i> sequencing) and total protein profiling assays (SDS-PAGE) techniques were used to detect genetic stability in micropropagated fig plantlets. <b>Results:</b> The Scorable PCR bands were produced with 10 SCoT primers used, where the total number of bands was 135 bands. Twenty polymorphic bands were generated with 18.4% of a polymorphism percentage. According to the result, no visual unique bands were generated which confirmed the genetic homogeneity of micropropagated plantlets samples compared to the control sample (mother plant). Sequence analysis and phylogenetic tree generated using fig <i>rpoC1</i> sequence showed high similarity between control and plantlets samples of fig plant. The protein profiling results revealed no remarkable changes between micropropagated plantlets and the mother plant. <b>Conclusion:</b> The results indicate that using SCoT, DNA barcoding and protein profiling have demonstrated their utility to detect genetic homogeneity in micropropagated fig plantlets, which suggests using of micropropagation protocol of plants applied on the plantlets in the current study as a reliable protocol for <i>in vitro</i> culture and conservation of fig plant.

摘要

背景与目的

体外繁殖是最大限度地增加植物物种多样性的可能方法之一。本研究旨在评估微繁殖的无花果树苗的遗传稳定性,并确定体外繁殖对沙特无花果树基因组含量的影响。

材料与方法

采用起始密码子靶向(SCoT)、叶绿体基因 RNA 聚合酶 1(rpoC1)测序和总蛋白谱分析(SDS-PAGE)技术检测微繁殖无花果树苗的遗传稳定性。

结果

使用 10 个 SCoT 引物产生了可评分的 PCR 带,总带数为 135 条。产生了 20 个多态性带,多态性百分比为 18.4%。结果表明,与对照样品(母株)相比,微繁殖苗样品没有产生可见的独特带,这证实了微繁殖苗样品的遗传同质性。基于 fig rpoC1 序列生成的序列分析和系统发育树显示,对照和无花果树苗样品之间具有高度相似性。蛋白谱分析结果表明,微繁殖苗与母株之间没有明显变化。

结论

结果表明,使用 SCoT、DNA 条形码和蛋白谱分析已证明其在检测微繁殖无花果树苗遗传同质性方面的有效性,这表明在当前研究中应用于植物的微繁殖方案作为无花果树苗的体外培养和保护的可靠方案。

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