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甜葉菊中新KAH基因的克隆及表達(dá)模式分析

發(fā)布時(shí)間:2018-04-15 18:58

  本文選題:甜葉菊 + 甜菊糖苷 ; 參考:《生物技術(shù)通報(bào)》2017年04期


【摘要】:為進(jìn)一步揭示甜菊糖苷生物合成途徑的分子調(diào)控機(jī)制,探究甜菊糖苷在甜葉菊葉片中積累差異原因。從不同甜葉菊材料中分別克隆關(guān)鍵分支點(diǎn)酶KAH的編碼基因,并對(duì)獲得的核苷酸序列進(jìn)行生物學(xué)功能分析和預(yù)測(cè)。結(jié)果顯示,最終共獲得6條高度同源的核苷酸序列,個(gè)別關(guān)鍵堿基的突變?cè)斐尚蛄虚_放讀碼框位置和大小的顯著差異,共預(yù)測(cè)獲得7個(gè)不同KAH編碼基因。KAH1、KAH2同時(shí)存在于SR1、SR3、鑫豐3號(hào)、譜星1號(hào)和守田3號(hào)中,KAH3存在于江甜1號(hào)和守田3號(hào)中,KAH4存在于江甜3號(hào)中,KAH5、KAH6、KAH7均存在于SR2中且Sr KAHs在甜菊糖苷積累多的品種和組織部位中表達(dá)更高。7個(gè)KAH基因均含有P450保守結(jié)構(gòu)域且均屬于CYP716家族成員;亞細(xì)胞定位預(yù)測(cè)KAH2和KAH4定位于細(xì)胞質(zhì)其他5個(gè)均定位在葉綠體中。甜葉菊中KAH編碼基因的數(shù)目、氨基酸長(zhǎng)度及在亞細(xì)胞結(jié)構(gòu)的定位差異或許是導(dǎo)致甜葉菊不同品種和組織中Sr KAHs表達(dá)和功能差異的主要原因并最終糖苷的積累差異。
[Abstract]:In order to further reveal the molecular regulation mechanism of stevioside biosynthesis pathway, the reason of stevioside accumulation difference in stevia leaves was investigated.The coding genes of key branching enzyme KAH were cloned from different stevia materials, and the obtained nucleotide sequences were analyzed and predicted.The results showed that a total of 6 highly homologous nucleotide sequences were obtained. The mutation of some key bases resulted in significant differences in the position and size of the open reading frame.It is predicted that seven different KAH coding genes. KAH1 and KAH2 exist simultaneously in SR1, SR3, Xinfeng 3, Xinfeng 3, Xinfeng 3, Xinfeng 3.Peixing 1 and Shutian 3 were found in Jiangtian 1 and Shutian 3 respectively. Both of them were found in SR2, and Sr KAHs were expressed in the cultivars and tissues with more stevioside accumulation. All of the 7 KAH genes were expressed in all of them.It contains P450 conserved domain and belongs to CYP716 family.Subcellular localization predicted that KAH2 and KAH4 were located in the cytoplasm and the other 5 were located in chloroplasts.The differences in the number of KAH coding genes, amino acid length and subcellular structure in stevia may be the main reasons leading to the difference of Sr KAHs expression and function in different varieties and tissues of stevia, and ultimately the difference of glycoside accumulation.
【作者單位】: 江蘇省農(nóng)科院經(jīng)濟(jì)作物研究所;
【基金】:江蘇省農(nóng)業(yè)三新工程[SXGC(2015)213] 江蘇省農(nóng)業(yè)科學(xué)院科技服務(wù)項(xiàng)目[KF(16)1042]
【分類號(hào)】:S566.9;Q943.2
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本文編號(hào):1755369

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