首页 > 分享 > 滇牡丹PdMYB57基因克隆及功能验证

滇牡丹PdMYB57基因克隆及功能验证

花匠小妙招
2024-11-25 14:28

滇牡丹PdMYB57基因克隆及功能验证

作者单位:

1.西南林业大学林学院;2.西南林业大学生物多样性保护学院;3.西南林业大学园林园艺学院

摘要 | | 访问统计 | 参考文献 [42] | | || 文章评论

摘要:

摘要【目的】滇牡丹（Paeonia delavayi，P.delavayi）作为珍贵的牡丹育种材料，具有丰富的花色资源，研究其MYB转录因子对花色的调控作用可为牡丹花色的分子育种工作提供帮助。【方法】以黄色及红色花滇牡丹花部组织为材料，进行徒手切片观察花青素在其细胞中的分布特征，并以2种花色滇牡丹材料的转录组数据为基础，通过本地Blast序列比对得到1个MYB转录因子PdMYB57。经基因克隆、生物信息学分析、系统进化树构建及同源序列比对对PdMYB57转录因子的理化性质及功能进行预测，再利用定量逆转录聚合酶链式反应（qPCR）、瞬时表达及高效液相色谱（HPLC）等方法对PdMYB57基因的功能进行验证。【结果】PdMYB57基因具有1个798 bp的完整开放阅读框，编码265个氨基酸，是定位于细胞核的亲水性不稳定蛋白；与拟南芥R2R3MYB家族中的SG6亚族以及卵叶牡丹（P.qiui）PqMYB113、牡丹（P.suffruticosa）PsMYB57/PsMYB58以及葡萄（Vitis vinifera，V vinifera）VvMYBA1/VvMYBA2等MYB转录因子聚为一簇，并具有典型的R2R3保守结构域[R/K]Px[P/A/R]xx[F/Y]基序；qPCR实验表明PdMYB57基因在红色花滇牡丹萼片及黄色花滇牡丹叶片中具有较高的表达量，在其他组织中表达量极低，几乎不表达，这与转录组数据结果一致；PdMYB57基因瞬时表达后能使烟草叶片产生紫色，HPLC检测表明PdMYB57基因瞬时表达的烟草叶片中含有矢车菊素3-O-芸香糖苷（Cyanidin- 3-O-rutinoside，Cy3R），而CK对照的烟草叶片没有产生颜色，其提取液中也未检测到Cy3R。【结论】PdMYB57基因编码1个R2R3MYB转录因子，为定位于细胞核中的亲水性不稳定蛋白，其表达具有组织特异性，并能促进植物花青素的合成。

Abstract:

Abstract [Objective] As a precious peony breeding material, Paeonia delavayi has abundant flower color resources. Studying the regulatory effect of its MYB transcription factors on flower color can provide assistance for molecular breeding of peony flower color. [Methods] Using the flower tissue of yellow and red P.delavayi as materials, observing the distribution characteristics of anthocyanins in its cells through hand sectioning. Based on the transcriptome data of the two materials, a MYB transcription factor PdMYB57 was obtained through sequence alignment. The physicochemical properties and functions of the PdMYB57 transcription factor were predicted through gene cloning, bioinformatics analysis, phylogenetic tree, and homologous sequence alignment. Subsequently, Quantitative reverse transcription polymerase chain reaction (qPCR), transient expression in tobacco, and High performance liquid chromatography (HPLC) were used to validate the function of PdMYB57. [Results] The PdMYB57 gene has a complete open reading frame of 798 bp, encoding 265 amino acids, and is located in the nucleus as a hydrophilic unstable protein; Cluster together with the SG6 subfamily in the R2R3MYB family of Arabidopsis, as well as MYB transcription factors that promote anthocyanin synthesis in P.qiui PqMYB113,P.suffruticosa PsMYB57/PsMYB58 and Vitis vinifera VvMYBA1/VvMYBA2, and exhibit a typical R2R3 conserved domain [R/K] Px [P/A/R] xx [F/Y] motif; the qPCR results showed that the PdMYB57 gene has high expression levels in the sepals of red P. delavayi and leaves of yellow P. delavayi. In other tissues, the expression level is extremely low and almost non-existent, which is consistent with the transcriptome data results; HPLC detection showed that the tobacco leaves with transient expression of the PdMYB57 gene contain Cyanidin- 3-O-rutinoside (Cy3R) , while the tobacco leaves with CK control did not produce color and Cy3R was not detected in the extraction solution. [Conclusion] The PdMYB57 gene encodes an R2R3-MYB transcription factor, located in the nucleus as a hydrophilic unstable protein. Its expression is tissue-specific and promotes the synthesis of anthocyanins in plants.

引用本文

杜春,平怀磊,刘思淇,等.滇牡丹PdMYB57基因克隆及功能验证[J].西北植物学报,2024,44(10):1577-1588

复制

分享文章指标点击次数:98 下载次数: 2456 HTML阅读次数: 14 历史收稿日期:2024-03-05 最后修改日期:2024-05-27 录用日期:2024-05-28 在线发布日期: 2024-08-21 文章二维码