1-Hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (IDS) is encoded by multicopy genes in gymnosperms Ginkgo biloba and Pinus taeda
1-Hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (IDS) is encoded by multicopy genes in gymnosperms Ginkgo biloba and Pinus taeda
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DOI：
10.1007/s00425-007-0616-x
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发表时间：
2008-01-01
期刊：
影响因子：
4.3
通讯作者：
Kim, Soo-Un
中科院分区：
文献类型：
--
作者：
Kim, Sang-Min;Kuzuyama, Tomohisa;Kim, Soo-Un
类异戊二烯是通过五碳中间体异戊烯基二磷酸（IPP）和二甲基烯丙基二磷酸（DMAPP）的缩合合成的，这些中间体来源于两种不同的生物合成途径：细胞溶质甲羟戊酸（MVA）和质体2-C-甲基-D-β 4-磷酸（MEP）途径。从裸子植物银杏（GbIDS 1、GbIDS 2和GbIDS 2 -1）和火炬松（PtIDS 1和PtIDS 2）中克隆了催化MEP途径最后一步的1-羟基-2-甲基-2-（E）-丁烯基4-二磷酸还原酶（IDS; EC 1.17.1.2）的多拷贝基因，并对其进行了表征。与其他植物入侵检测系统构建的系统发育树表明裸子植物入侵检测系统与被子植物入侵检测系统存在明显差异。裸子植物IDS分支包含两个亚分支，一个由GbIDS 1和PtIDS 1组成，另一个由GbIDS 2、GbIDS 2 -1和PtIDS 2组成。G.银杏IDS，除了GbIDS 2 -1，成功地补充大肠杆菌DLYT 1，lytB破坏，确认在体内的能力的同工酶。在为期4周的研究期间，虽然GbIDS 1在培养的根和叶中的转录水平相似，但GbIDS 1在培养的根和叶中的转录水平相似。GbIDS 2的转录本主要存在于银杏胚的根中，银杏内酯在根中生物合成。PtIDS 2在二萜类树脂产生木材中的转录水平比在其他组织中的转录水平高4-5倍。更高水平的GbIDS 1转录诱导光，而那些GbIDS 2茉莉酸甲酯治疗增加。这些结果强烈暗示GbIDS 2和PtIDS 2与次生代谢具有高度相关性。在拟南芥瞬时表达系统中，GbIDS 1的N端100个氨基酸残基将融合的GFP蛋白递送到叶绿体、细胞质和细胞核中，而GbIDS 2、GbIDS 2 -1和两个PtIDSs的N端100个氨基酸残基仅将GFP递送到叶绿体中。
Isoprenoids are synthesized through the condensation of five-carbon intermediates, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), derived from two distinct biosynthetic routes: cytosolic mevalonate (MVA) and plastidial 2-C-methyl-D-erythritol 4-phosphate (MEP) pathways. 1-Hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (IDS; EC 1.17.1.2), which catalyzes the last step of MEP pathway, was cloned as a multicopy gene from gymnosperms Ginkgo biloba (GbIDS1, GbIDS2, and GbIDS2-1) and Pinus taeda (PtIDS1 and PtIDS2), and characterized. Phylogenetic tree constructed with other plant IDSs demonstrated gymnosperm IDSs were distinctively different from angiosperm IDSs. The gymnosperm IDS clade contained two subclades, one composed of GbIDS1 and PtIDS1, and the other composed of GbIDS2, GbIDS2-1, and PtIDS2. G. biloba IDSs, except GbIDS2-1, successfully complemented Escherichia coli DLYT1, a lytB disruptant, confirming the in vivo competency of isozymes. During the 4 weeks study period, although transcript levels of GbIDS1s were similar both in roots and leaves of cultured G. biloba embryo, the transcripts of GbIDS2 predominantly occurred in the embryo roots, where diterpene ginkgolides are biosynthesized. Levels of PtIDS2 transcripts in the diterpenoid resin-producing wood were 4-5 times higher than those in other tissues. Higher levels of GbIDS1 transcripts were induced by light, whereas those of GbIDS2 were increased by methyl jasmonate treatment. These results strongly imply GbIDS2 and PtIDS2 have high correlation with secondary metabolism. In Arabidopsis transient expression system, N-terminal 100 amino acid residues of GbIDS1 delivered fused GFP protein into chloroplast as well as cytosol and nucleus, whereas those of GbIDS2, GbIDS2-1, and two PtIDSs delivered GFP only into chloroplast.