桃硬质性状可能源于PpYUC11基因启动子区域CACTA型转座子的插入
发布时间：2018-08-30 18:22
【摘要】：【目的】溶质(melting flesh,MF)型桃果实的成熟软化依赖于生长素诱导的乙烯合成相关基因的表达,硬质(stony-hard,SH)型桃不软化的原因是其在成熟期IAA的合成受阻导致乙烯不能正常释放。YUCCA编码类黄素单加氧酶(flavin monooxygenase-like),催化吲哚丙酮酸合成IAA。前期研究发现,PpYUC11为控制桃SH肉质性状候选基因,笔者在此基础上继续探讨PpYUC11基因在溶质型和硬质型桃果实中差异表达的原因。【方法】采用实时荧光定量PCR分析其在果实和种子的表达特性,扩增SH和非SH型桃品种PpYUC11基因的上游区域,以期明确该基因在不同桃品种类型的序列特征及时空表达特征。【结果】PpYUC11基因在MF型桃果实成熟期果肉中的表达丰度高于其在SH型桃中的表达丰度,而在2种肉质类型种子中的表达基本没有差异,比较PpYUC11基因的上游调控区域的序列发现,SH型桃相比MF型桃,PpYUC11基因ATG上游约2.1 kb处存在大片段的插入。核苷酸比对发现该片段预测为CACTA型DNA转座子片段,同时基于转座子的标记可以明显区分SH和非SH类型,调控顺式元件分析表明PpYUC11启动子存在多种激素响应元件,其中在插入位点上游100 bp左右存在1个果实特异性元件,暗示该元件参与PpYUC11的组织特异性表达,而转座子的插入可能导致该元件不能与果实成熟相关的转录因子正常结合。【结论】桃SH肉质性状可能由CACTA型DNA转座子插入PpYUC11基因启动子区域所致,本试验不仅为后期选育和鉴定SH型品种提供了可靠的分子标记,同时为进一步解析SH型桃果实的突变机制奠定基础。
[Abstract]:[objective] the maturation and softening of solute (melting flesh,MF) peach fruit depends on the expression of auxin induced ethylene biosynthesis related genes. The reason why stony-hard,SH peach does not soften is that the synthesis of IAA in mature peach is blocked and ethylene can not be released normally. Yucca encodes Flavin monooxygenase (flavin monooxygenase-like) and catalyzes the synthesis of IAA. from indopyruvate. In previous studies, PpYUC11 was found to be a candidate gene for controlling SH fleshy traits in peach. On this basis, the reasons of differential expression of PpYUC11 gene in solute and hard peach fruits were investigated. [methods] Real-time quantitative PCR was used to analyze the expression characteristics of PpYUC11 gene in fruits and seeds. The upstream region of PpYUC11 gene in SH and non-SH peach cultivars was amplified. [results] the expression abundance of PpYUC11 gene in the flesh of MF peach was higher than that in SH type peach. However, there was no difference in expression between the two types of fleshy seeds. Comparing the sequence of upstream regulatory region of PpYUC11 gene, it was found that there was a large fragment insertion in SH-type peach compared with MF type PpYUC11 gene ATG about 2.1 kb upstream. Nucleotide alignment revealed that the fragment was predicted to be CACTA type DNA transposon fragment, and the SH and non-SH types could be distinguished by transposon markers. The analysis of regulatory cis-element showed that there were many hormone response elements in PpYUC11 promoter. There was a fruit specific element about 100 bp upstream of the insertion site, which implied that the element was involved in the tissue specific expression of PpYUC11. The insertion of transposon may result in the element not being able to bind normally with transcription factors related to fruit ripening. [conclusion] the fleshy character of SH in peach may be caused by the insertion of CACTA type DNA transposon into the promoter region of PpYUC11 gene. This experiment not only provided reliable molecular markers for later selection and identification of SH type varieties, but also laid a foundation for further analysis of mutation mechanism of SH type peach fruit.
【作者单位】： 中国农业科学院郑州果树研究所·国家桃葡萄改良中心·农业部果树育种技术重点实验室;
【基金】：国家自然科学基金(31501732) 中国农业科学院科技创新工程(CAAS-ASTIP-2017-ZFRI) 河南省科技厅基础前沿项目(152300410137)
【分类号】：S662.1
本文编号：2213914
[Abstract]:[objective] the maturation and softening of solute (melting flesh,MF) peach fruit depends on the expression of auxin induced ethylene biosynthesis related genes. The reason why stony-hard,SH peach does not soften is that the synthesis of IAA in mature peach is blocked and ethylene can not be released normally. Yucca encodes Flavin monooxygenase (flavin monooxygenase-like) and catalyzes the synthesis of IAA. from indopyruvate. In previous studies, PpYUC11 was found to be a candidate gene for controlling SH fleshy traits in peach. On this basis, the reasons of differential expression of PpYUC11 gene in solute and hard peach fruits were investigated. [methods] Real-time quantitative PCR was used to analyze the expression characteristics of PpYUC11 gene in fruits and seeds. The upstream region of PpYUC11 gene in SH and non-SH peach cultivars was amplified. [results] the expression abundance of PpYUC11 gene in the flesh of MF peach was higher than that in SH type peach. However, there was no difference in expression between the two types of fleshy seeds. Comparing the sequence of upstream regulatory region of PpYUC11 gene, it was found that there was a large fragment insertion in SH-type peach compared with MF type PpYUC11 gene ATG about 2.1 kb upstream. Nucleotide alignment revealed that the fragment was predicted to be CACTA type DNA transposon fragment, and the SH and non-SH types could be distinguished by transposon markers. The analysis of regulatory cis-element showed that there were many hormone response elements in PpYUC11 promoter. There was a fruit specific element about 100 bp upstream of the insertion site, which implied that the element was involved in the tissue specific expression of PpYUC11. The insertion of transposon may result in the element not being able to bind normally with transcription factors related to fruit ripening. [conclusion] the fleshy character of SH in peach may be caused by the insertion of CACTA type DNA transposon into the promoter region of PpYUC11 gene. This experiment not only provided reliable molecular markers for later selection and identification of SH type varieties, but also laid a foundation for further analysis of mutation mechanism of SH type peach fruit.
【作者单位】： 中国农业科学院郑州果树研究所·国家桃葡萄改良中心·农业部果树育种技术重点实验室;
【基金】：国家自然科学基金(31501732) 中国农业科学院科技创新工程(CAAS-ASTIP-2017-ZFRI) 河南省科技厅基础前沿项目(152300410137)
【分类号】：S662.1
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