【摘要】：【目的】溶質(zhì)(melting flesh,MF)型桃果實(shí)的成熟軟化依賴于生長(zhǎng)素誘導(dǎo)的乙烯合成相關(guān)基因的表達(dá),硬質(zhì)(stony-hard,SH)型桃不軟化的原因是其在成熟期IAA的合成受阻導(dǎo)致乙烯不能正常釋放。YUCCA編碼類黃素單加氧酶(flavin monooxygenase-like),催化吲哚丙酮酸合成IAA。前期研究發(fā)現(xiàn),PpYUC11為控制桃SH肉質(zhì)性狀候選基因,筆者在此基礎(chǔ)上繼續(xù)探討PpYUC11基因在溶質(zhì)型和硬質(zhì)型桃果實(shí)中差異表達(dá)的原因�！痉椒ā坎捎脤�(shí)時(shí)熒光定量PCR分析其在果實(shí)和種子的表達(dá)特性,擴(kuò)增SH和非SH型桃品種PpYUC11基因的上游區(qū)域,以期明確該基因在不同桃品種類型的序列特征及時(shí)空表達(dá)特征。【結(jié)果】PpYUC11基因在MF型桃果實(shí)成熟期果肉中的表達(dá)豐度高于其在SH型桃中的表達(dá)豐度,而在2種肉質(zhì)類型種子中的表達(dá)基本沒(méi)有差異,比較PpYUC11基因的上游調(diào)控區(qū)域的序列發(fā)現(xiàn),SH型桃相比MF型桃,PpYUC11基因ATG上游約2.1 kb處存在大片段的插入。核苷酸比對(duì)發(fā)現(xiàn)該片段預(yù)測(cè)為CACTA型DNA轉(zhuǎn)座子片段,同時(shí)基于轉(zhuǎn)座子的標(biāo)記可以明顯區(qū)分SH和非SH類型,調(diào)控順式元件分析表明PpYUC11啟動(dòng)子存在多種激素響應(yīng)元件,其中在插入位點(diǎn)上游100 bp左右存在1個(gè)果實(shí)特異性元件,暗示該元件參與PpYUC11的組織特異性表達(dá),而轉(zhuǎn)座子的插入可能導(dǎo)致該元件不能與果實(shí)成熟相關(guān)的轉(zhuǎn)錄因子正常結(jié)合�！窘Y(jié)論】桃SH肉質(zhì)性狀可能由CACTA型DNA轉(zhuǎn)座子插入PpYUC11基因啟動(dòng)子區(qū)域所致,本試驗(yàn)不僅為后期選育和鑒定SH型品種提供了可靠的分子標(biāo)記,同時(shí)為進(jìn)一步解析SH型桃果實(shí)的突變機(jī)制奠定基礎(chǔ)。
[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.
【作者單位】： 中國(guó)農(nóng)業(yè)科學(xué)院鄭州果樹研究所·國(guó)家桃葡萄改良中心·農(nóng)業(yè)部果樹育種技術(shù)重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金(31501732) 中國(guó)農(nóng)業(yè)科學(xué)院科技創(chuàng)新工程(CAAS-ASTIP-2017-ZFRI) 河南省科技廳基礎(chǔ)前沿項(xiàng)目(152300410137)
【分類號(hào)】：S662.1

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