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半夏的从头测序与转录组分析鉴定参与苯甲酸和麻黄碱生物合成的候选基因。

De novo Sequencing and Transcriptome Analysis of Pinellia ternata Identify the Candidate Genes Involved in the Biosynthesis of Benzoic Acid and Ephedrine.

作者信息

Zhang Guang-Hui, Jiang Ni-Hao, Song Wan-Ling, Ma Chun-Hua, Yang Sheng-Chao, Chen Jun-Wen

机构信息

Yunnan Research Center on Good Agricultural Practice for Dominant Chinese Medicinal Materials, Yunnan Agricultural University Kunming, China.

出版信息

Front Plant Sci. 2016 Aug 16;7:1209. doi: 10.3389/fpls.2016.01209. eCollection 2016.

Abstract

BACKGROUND

The medicinal herb, Pinellia ternata, is purported to be an anti-emetic with analgesic and sedative effects. Alkaloids are the main biologically active compounds in P. ternata, especially ephedrine that is a phenylpropylamino alkaloid specifically produced by Ephedra and Catha edulis. However, how ephedrine is synthesized in plants is uncertain. Only the phenylalanine ammonia lyase (PAL) and relevant genes in this pathway have been characterized. Genomic information of P. ternata is also unavailable.

RESULTS

We analyzed the transcriptome of the tuber of P. ternata with the Illumina HiSeq™ 2000 sequencing platform. 66,813,052 high-quality reads were generated, and these reads were assembled de novo into 89,068 unigenes. Most known genes involved in benzoic acid biosynthesis were identified in the unigene dataset of P. ternata, and the expression patterns of some ephedrine biosynthesis-related genes were analyzed by reverse transcription quantitative real-time PCR (RT-qPCR). Also, 14,468 simple sequence repeats (SSRs) were identified from 12,000 unigenes. Twenty primer pairs for SSRs were randomly selected for the validation of their amplification effect.

CONCLUSION

RNA-seq data was used for the first time to provide a comprehensive gene information on P. ternata at the transcriptional level. These data will advance molecular genetics in this valuable medicinal plant.

摘要

背景

草药半夏据称是一种具有镇痛和镇静作用的止吐药。生物碱是半夏的主要生物活性化合物,尤其是麻黄碱,它是麻黄属植物和巧茶特有的苯基丙胺类生物碱。然而,植物中麻黄碱的合成方式尚不确定。该途径中仅苯丙氨酸解氨酶(PAL)及相关基因已得到表征。半夏的基因组信息也尚未可得。

结果

我们使用Illumina HiSeq™ 2000测序平台分析了半夏块茎的转录组。共生成了66,813,052条高质量 reads,并将这些reads从头组装成89,068个单基因。在半夏的单基因数据集中鉴定出了大多数参与苯甲酸生物合成的已知基因,并通过逆转录定量实时PCR(RT-qPCR)分析了一些与麻黄碱生物合成相关基因的表达模式。此外,从12,000个单基因中鉴定出了14,468个简单序列重复(SSR)。随机选择了20对SSR引物来验证其扩增效果。

结论

首次使用RNA-seq数据在转录水平上提供了关于半夏的全面基因信息。这些数据将推动这种珍贵药用植物的分子遗传学研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34cd/4986801/a1b0797a547b/fpls-07-01209-g0001.jpg

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