Duan Hui, Huang Meng-Yu, Palacio Kathryn, Schuler Mary A
Department of Cell and Developmental Biology, University of Illinois, Urbana, IL 61801, USA.
Plant Physiol. 2005 Nov;139(3):1529-44. doi: 10.1104/pp.105.067249. Epub 2005 Oct 28.
The CYP74B2 gene in Arabidopsis (Arabidopsis thaliana) ecotype Columbia (Col) contains a 10-nucleotide deletion in its first exon that causes it to code for a truncated protein not containing the P450 signature typical of other CYP74B subfamily members. Compared to CYP74B2 transcripts in the Landsberg erecta (Ler) ecotype that code for full-length hydroperoxide lyase (HPL) protein, CYP74B2 transcripts in the Col ecotype accumulate at substantially reduced levels. Consistent with the nonfunctional HPL open reading frame in the Col ecotype, in vitro HPL activity analyses using either linoleic acid hydroperoxide or linolenic acid hydroperoxide as substrates show undetectable HPL activity in the Col ecotype and C6 volatile analyses using leaf homogenates show substantially reduced amounts of hexanal and no detectable trans-2-hexenal generated in the Col ecotype. P450-specific microarrays and full-genome oligoarrays have been used to identify the range of other transcripts expressed at different levels in these two ecotypes potentially as a result of these variations in HPL activity. Among the transcripts expressed at significantly lower levels in Col leaves are those coding for enzymes involved in the synthesis of C6 volatiles (LOX2, LOX3), jasmonates (OPR3, AOC), and aliphatic glucosinolates (CYP83A1, CYP79F1, AOP3). Two of the three transcripts coding for aliphatic glucosinolates (CYP83A1, AOP3) are also expressed at significantly lower levels in Col flowers.
拟南芥生态型哥伦比亚(Col)中的CYP74B2基因在其第一个外显子中存在10个核苷酸的缺失,这导致它编码一种截短的蛋白质,该蛋白质不包含其他CYP74B亚家族成员典型的P450特征。与编码全长氢过氧化物裂解酶(HPL)蛋白的直立叶兰(Ler)生态型中的CYP74B2转录本相比,Col生态型中的CYP74B2转录本积累水平大幅降低。与Col生态型中无功能的HPL开放阅读框一致,以亚油酸氢过氧化物或亚麻酸氢过氧化物为底物的体外HPL活性分析表明,Col生态型中未检测到HPL活性,使用叶片匀浆的C6挥发性分析表明,Col生态型中己醛的量大幅减少,且未检测到反式-2-己烯醛的产生。P450特异性微阵列和全基因组寡核苷酸阵列已被用于确定这两种生态型中由于HPL活性的这些变化可能在不同水平表达的其他转录本的范围。在Col叶片中表达水平显著较低的转录本中,有那些编码参与C6挥发性物质(LOX2、LOX3)、茉莉酸(OPR3、AOC)和脂肪族芥子油苷(CYP83A1、CYP79F1、AOP3)合成的酶的转录本。编码脂肪族芥子油苷的三个转录本中的两个(CYP83A1、AOP3)在Col花中的表达水平也显著较低。