Differential hydroxylation efficiency of the two non-heme carotene hydroxylases: DcBCH1, rather than DcBCH2, plays a major role in carrot taproot.
Differential hydroxylation efficiency of the two non-heme carotene hydroxylases: DcBCH1, rather than DcBCH2, plays a major role in carrot taproot.
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两种非血红素胡萝卜素羟化酶的羟基化效率不同：DcBCH1（而不是 DcBCH2）在胡萝卜主根中起主要作用
DOI：
10.1093/hr/uhac193
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发表时间：
2022
影响因子：
8.7
通讯作者：
中科院分区：
文献类型：
--
作者：
摘要 胡萝卜素羟化酶在催化胡萝卜素羟基化为叶黄素方面发挥着重要作用，包括非血红素胡萝卜素羟化酶（BCH型）和含血红素细胞色素P450羟化酶（P450型）两种类型。胡萝卜基因组中注释了两个 BCH 编码基因。然而，BCH 的作用以及胡萝卜中重复的 BCH 之间是否存在功能相互作用仍不清楚。在这项研究中，从胡萝卜中克隆了两个 BCH 编码基因 DcBCH1 和 DcBCH2。在不同胡萝卜素积累水平的胡萝卜主根中，DcBCH1的相对表达量显着高于DcBCH2。 “KRD”（高胡萝卜素积累）胡萝卜中 DcBCH1 的过度表达使主根颜色从橙色变为黄色，同时 α-胡萝卜素和 β-胡萝卜素大幅减少。过表达 DcBCH2 的转基因‘KRD’胡萝卜和对照胡萝卜之间主根颜色没有明显变化。同时，过表达DcBCH2的胡萝卜主根中α-胡萝卜素含量下降，但β-胡萝卜素含量与对照胡萝卜相比没有显着变化。使用 CRISPR/Cas9 系统敲除“KRD”胡萝卜中的 DcBCH1，使主根颜色从橙色变为粉橙色；与对照胡萝卜相比，主根中α-胡萝卜素含量略有增加，而β-胡萝卜素含量仍显着降低。在 DcBCH1 敲除胡萝卜中，DcBCH2 的转录水平显着增加。这些结果表明，在胡萝卜主根中，DcBCH1发挥了BCH酶的主要功能，可羟基化α-胡萝卜素和β-胡萝卜素； DcBCH1和DcBCH2具有功能冗余，这两个DcBCH可以部分互补。
Abstract Carotene hydroxylase plays an important role in catalyzing the hydroxylation of carotene to xanthopylls, including two types: non-heme carotene hydroxylase (BCH type) and heme-containing cytochrome P450 hydroxylase (P450 type). Two BCH-encoding genes were annotated in the carrot genome. However, the role of BCHs and whether there are functional interactions between the duplicated BCHs in carrot remains unclear. In this study, two BCH encoding genes, DcBCH1 and DcBCH2, were cloned from carrot. The relative expression level of DcBCH1 was much higher than that of DcBCH2 in carrot taproots with different carotene accumulation levels. Overexpression of DcBCH1 in ‘KRD’ (high carotene accumulated) carrot changed the taproot color from orange to yellow, accompanied by substantial reductions in α-carotene and β-carotene. There was no obvious change in taproot color between transgenic ‘KRD’ carrot overexpressing DcBCH2 and control carrot. Simultaneously, the content of α-carotene in the taproot of DcBCH2-overexpressing carrot decreased, but the content of β-carotene did not change significantly in comparison with control carrot. Using the CRISPR/Cas9 system to knock out DcBCH1 in ‘KRD’ carrot lightened the taproot color from orange to pink-orange; the content of α-carotene in the taproot increased slightly, while the β-carotene content was still significantly decreased, compared with control carrot. In DcBCH1-knockout carrot, the transcript level of DcBCH2 was significantly increased. These results indicated that in carrot taproot, DcBCH1 played the main function of BCH enzyme, which could hydroxylate α-carotene and β-carotene; DcBCH1 and DcBCH2 had functional redundancy, and these two DcBCHs could partially compensate for each other.
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影响因子：
4.9
作者：
Kim, Joonyul;Smith, James J.;DellaPenna, Dean
通讯作者：
DellaPenna, Dean
影响因子：
5.3
作者：
Diretto, Gianfranco;Welsch, Ralf;Tavazza, Raffaela;Mourgues, Fabienne;Pizzichini, Daniele;Beyer, Peter;Giuliano, Giovanni
通讯作者：
Giuliano, Giovanni
影响因子：
7.4
作者：
Du, Hao;Wang, Nili;Xiong, Lizhong
通讯作者：
Xiong, Lizhong
DOI：
10.1073/pnas.241374598
发表时间：
2002-01-22
影响因子：
11.1
作者：
Fraser, PD;Romer, S;Bramley, PM
通讯作者：
Bramley, PM
影响因子：
3.8
作者：
Kim, Sun Ha;Ahn, Young Ock;Kwak, Sang-Soo
通讯作者：
Kwak, Sang-Soo