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两种植食豆属植物叶绿体基因组的测序、比较分析与分子标记开发。

Chloroplast genomes of two Pueraria DC. species: sequencing, comparative analysis and molecular marker development.

机构信息

Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

Institute of Chinese Medicinal Materials, Hubei Academy of Agricultural Sciences, Enshi, China.

出版信息

FEBS Open Bio. 2022 Feb;12(2):349-361. doi: 10.1002/2211-5463.13335. Epub 2021 Dec 26.

Abstract

Puerariae lobatae radix (Ge-Gen in Chinese) and Puerariae thomsonii radix (Fen-Ge) are widely used as medicine and health products, particularly in Chinese medicine. Puerarin and daidzein are the primary bioactive compounds in Puerariae radix. These isoflavones have been used to treat cardiovascular and cerebrovascular diseases, hypertension, diabetes, and osteoporosis. The content of puerarin in Ge-Gen is about six times higher than that in Fen-Ge, so its use has a higher pharmacological effect. It is therefore of great importance to effectively distinguish between these two species. However, because their basal plants, P. lobata (Willd.) Ohwi and P. thomsonii Benth., possess an extremely similar appearance, and detecting the level of chemical constituents is just a rough distinction, it is necessary to develop more efficient identification approaches. Here the complete chloroplast genomes of P. lobata and P. thomsonii were deciphered, including sequencing, assembly, comparative analysis, and molecular marker development. The results showed that they are 153,393 and 153,442 bp in length, respectively; both contain 124 annotated genes, including eight encoding rRNA, 29 encoding tRNA, and 87 encoding proteins. Phylogenetic analysis showed that they form a clade, indicating that they originate from the same ancestor. After obtaining 10 intergenic/intronic regions with a genetic distance greater than 0.5 cm, primers were designed to amplify regions of high variability in P. lobata and P. thomsonii. Finally, a 60-bp differential base fragment, located in the intron of rpl16, was developed as a molecular marker to efficiently distinguish between these two species.

摘要

葛(gen)和粉葛(fen-ge)是两种广泛应用于医药和保健品领域的豆科植物,尤其在中国传统医学中应用广泛。葛根素和大豆苷元是葛根中的主要生物活性化合物。这些异黄酮类化合物已被用于治疗心血管和脑血管疾病、高血压、糖尿病和骨质疏松症。葛(gen)中的葛根素含量约为粉葛(fen-ge)的六倍,因此其药用效果更高。因此,有效区分这两种植物具有重要意义。然而,由于它们的基生植物野葛(Pueraria lobata (Willd.) Ohwi)和粉葛(P. thomsonii Benth.)外观极为相似,且检测化学成分水平只是一种粗略的区分,因此需要开发更有效的鉴定方法。本研究中,我们破译了野葛和粉葛的完整叶绿体基因组,包括测序、组装、比较分析和分子标记开发。结果表明,它们的长度分别为 153,393 和 153,442bp,均包含 124 个注释基因,包括 8 个编码 rRNA、29 个编码 tRNA 和 87 个编码蛋白质。系统发育分析表明,它们形成一个分支,表明它们起源于同一祖先。获得了 10 个遗传距离大于 0.5cm 的基因间隔区/内含子后,我们设计了引物来扩增野葛和粉葛中高变异性的区域。最后,开发了一个 60bp 的差异碱基片段,位于 rpl16 的内含子中,作为一种分子标记,可有效区分这两种植物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/99fc/8804624/6ee048a2e148/FEB4-12-349-g007.jpg

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