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cDNA-AFLP 分析当归花蕾和芽-茎尖分生组织之间基因表达的差异。

cDNA-AFLP analysis of gene expression differences between the flower bud and sprout-shoot apical meristem of Angelica sinensis (Oliv.) Diels.

机构信息

Nanjing University of Chinese Medicine, Nanjing, China.

出版信息

Genet Mol Biol. 2011 Apr;34(2):274-9. doi: 10.1590/s1415-47572011000200018. Epub 2011 Apr 1.

Abstract

Angelica sinensis (Oliv.) Diels (Umbelliferae) is a well-known medicinal plant mainly distributed in Gansu Province of China. Its local and global demand is significant because of its food and medicinal applications. However, the early bolting rate of Angelica sinensis (Oliv.) Diels reaches 20%-60%, which seriously affects its food and medicinal qualities. Thus, differences in gene expression between the flower bud and sprout-shoot apical meristem underwent analysis, by means of cDNA-amplified restriction fragment length polymorphism, to better understand the flowering mechanism. 64 primer sets, each of which amplified to 60 transcript-derived fragments (TDFs), were used. Among these TDFs, 26 were expressed specifically in the flower bud. After cloning and sequencing, 32 distinct sequences were obtained from these 26 TDFs, and 25 were found with homologous sequences in databases. Confirmation of differential expression of 13 sequences was obtained by semi-quantitative RT-PCR, their showing higher expression levels in flower buds. These homologous sequences encode transposable elements, pentatricopeptide repeat-containing proteins, DNA-binding transcription factors, zinc finger (B-box type) family proteins, NADP-dependent sorbitol 6-phosphate dehydrogenase (S6PDH), amongst others.

摘要

当归(Angelica sinensis(Oliv.)Diels)(伞形科)是一种在中国甘肃省广泛分布的药用植物,因其具有食用和药用价值,其在中国和全球的需求量都很大。然而,当归的早期抽薹率高达 20%-60%,这严重影响了其食用和药用品质。因此,通过 cDNA 扩增限制片段长度多态性(cDNA-amplified restriction fragment length polymorphism)的方法,对花和芽顶端分生组织之间的基因表达差异进行了分析,以便更好地了解其开花机制。使用了 64 个引物对,每个引物对扩增到 60 个转录衍生片段(TDFs)。在这些 TDFs 中,有 26 个特异性地在花芽中表达。对这些 TDFs 进行克隆和测序后,从这 26 个 TDFs 中获得了 32 个不同的序列,其中 25 个在数据库中具有同源序列。通过半定量 RT-PCR 验证了 13 个序列的差异表达,这些序列在花芽中的表达水平更高。这些同源序列编码转座元件、五肽重复蛋白、DNA 结合转录因子、锌指(B-盒型)家族蛋白、NADP 依赖性山梨醇 6-磷酸脱氢酶(S6PDH)等。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f81/3115322/8f2d8cb67dae/gmb-34-2-274-gfig1.jpg

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