Subjects: Biology >> Botany >> Applied botany submitted time 2018-10-26 Cooperative journals: 《广西植物》
Abstract：海南龙血树是国产血竭的主要基源植物，其血竭主要化学成分为类黄酮化合物。目前对血竭的研究集中在化学成分及药理活性分析，对血竭形成的分子机制并不了解。WD40转录因子在类黄酮的积累中具有重要的调控作用。本研究根据海南龙血树转录组数据，利用RT-PCR技术在海南龙血树中克隆了一个WD40基因DcWD40-1，该基因包含一个1 353 bp的开放阅读框，编码450个氨基酸，蛋白质分子量50.77 kD，理论等电点5.71。生物信息学分析显示，DcWD40-1属于WD40蛋白家族成员，具有5个保守的WD40结构域，和其他植物WD40蛋白同源性高，保守性强。利用Genome Walking方法分离了1 503 bp的DcWD40-1启动子序列，该区域具有典型真核生物启动子结构特征，并含有多个应答激素和胁迫的响应元件。表达分析显示，血竭诱导剂能够诱导DcWD40-1的表达，DcWD40-1的变化与血竭形成及类黄酮积累正相关。此外，DcWD40-1也能对茉莉酸甲酯、细胞分裂素、油菜素内酯和UV-B处理做出积极响应。本结果为进一步研究DcWD40-1在类黄酮生物合成中的潜在功能和作用机制奠定基础。
Subjects: Agriculture, Forestry,Livestock & Aquatic Products Science >> Plant Protection submitted time 2018-07-18 Cooperative journals: 《广西植物》
Abstract： Anthocyanin-rich purple sweet potatos have high edible and medicinal value. Anthocyanin biosynthesis is controlled by structural genes and regulatory genes. The bHLH (basic helix-loop-helix protein) transcription factor plays an important role in anthocyanin biosynthesis by regulating multiple structural genes. However, there are no reports about bHLH regulating anthocyanin biosynthesis in sweet potato. In this study, a bHLH gene named IbGL3 was cloned in Ipomoea batatas based on transcriptome data and RT-PCR technology. The full-length cDNA of IbGL3 was 2 120 bp, containing 1 878 bp opening reading frame (ORF), and encoding 625 amino acids. The encoded protein of IbGL3 has a molecular weight of 69.08 kD and the theoretical isoelectric point (pI) of 5.20. The IbGL3 protein was classified as unstable protein instability with index of instability index 43.06, and belonged to hydrophilic protein with grand average of hydropathicity of -0.555. Subcellular localization prediction showed that IbGL3 was most likely to be located in the nucleus, and the nuclear localization signal (PSSKKRKASKT) was located at the C-terminal. Signal peptide analysis showed that IbGL3 protein had no signal peptide and was not a secretory protein. Transmembrane domain analysis showed that IbGL3 protein had no transmembrane domain. The IbGL3 protein had highly conserved MIR motif, bHLH domain and ACT domain, shared high identities and similar domains with bHLH proteins involved in anthocyanin biosynthesis from other plants. Phylogenetic analysis showed that and IbGL3 was clustered in the IIIf bHLH subgroup together with other anthocyanin-related bHLH proteins. The expression of IbGL3 in the storage root of different sweet potato varieties was detected by quantitative polymerase chain reaction (qPCR). The results indicated that IbGL3 was mainly expressed in purple-fleshed sweet potato, followed by light purple-fleshed sweet potato and weakly expressed in white-fleshed sweet potato, and its expression was positively related to the accumulation of anthocyanin Ipomoea batatas. The results showed IbGL3 may be involved in regulating anthocyanin biosynthesis in Ipomoea batatas. The results will provide assistance and guidance to further understand the function and molecular mechanism of IbGL3 in the biosynthesis of anthocyanins in sweet potato, and lay the foundation for genetic improvement of anthocyanin in sweet potato.
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