Identification of C4 responsive genes in the facultative C4 plant Hydrilla verticillata
Identification of C4 responsive genes in the facultative C4 plant Hydrilla verticillata
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DOI：
10.1007/s11120-006-9049-9
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发表时间：
2006-04
影响因子：
3.7
通讯作者：
Srinath K. Rao;H. Fukayama;J. Reiskind;M. Miyao;G. Bowes
中科院分区：
文献类型：
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作者：
Srinath K. Rao;H. Fukayama;J. Reiskind;M. Miyao;G. Bowes
水生单子叶植物黑藻Hydrilaverticillata（L.f.）Royle是一种有充分文献记载的兼性C4 NADP-苹果酸酶物种，其中C4和卡尔文循环在同一细胞中进行，特异性羧化酶分别局限于细胞质和叶绿体。几个关键的组成部分已经在分子水平上进行了表征，因此本研究的目的是开始，以确定其他的，不太明显的，可能是必要的功能性单细胞C4系统的元素。采用差异显示技术，从C3和C4 H的mRNA群体。筛选轮枝藻叶并比较表达谱。从这项研究中，分离出65个克隆，并进行定制的宏阵列分析;发现25个克隆在C4叶中上调。北方和半定量RT-PCR分析用于确认。从这些筛选中，鉴定了13个C4上调基因。其中一个编码一个以前公认的C4磷酸烯醇式丙酮酸羧化酶，和两个编码不同的丙酮酸正磷酸二激酶亚型，新的发现为H。轮生植物。编码转运蛋白、氨基转移酶和两种伴侣蛋白的基因也被上调。从北方/半定量RT-PCR分析中确定了12个假阳性，主要是管家基因。本研究中获得的序列数据列于dbEST数据库（DV 216698至DV 216767）中。作为一个缺乏Kranz解剖结构的单细胞C4系统，更好地了解H.轮枝孢属植物可能有助于在C3作物物种中设计转基因C4系统。
The aquatic monocotHydrilla verticillata(L.f.) Royle is a well-documented facultative C4NADP-malic enzyme species in which the C4and Calvin cycles operate in the same cell with the specific carboxylases confined to the cytosol and chloroplast, respectively. Several key components had already been characterized at the molecular level, thus the purpose of this study was to begin to identify other, less obvious, elements that may be necessary for a functional single-cell C4system. Using differential display, mRNA populations from C3and C4H. verticillataleaves were screened and expression profiles compared. From this study, 65 clones were isolated and subjected to a customized macroarray analysis; 25 clones were found to be upregulated in C4leaves. Northern and semi-quantitative RT-PCR analyses were used for confirmation. From these screenings, 13 C4upregulated genes were identified. Among these one encoded a previously recognized C4phosphoenolpyruvate carboxylase, and two encoded distinct pyruvate orthophosphate dikinase isoforms, new findings forH. verticillata. Genes that encode a transporter, an aminotransferase and two chaperonins were also upregulated. Twelve false positives, mostly housekeeping genes, were determined from the Northern/semi-quantitative RT-PCR analyses. Sequence data obtained in this study are listed in the dbEST database (DV216698 to DV216767). As a single-cell C4system that lacks Kranz anatomy, a better understanding of howH. verticillataoperates may facilitate the design of a transgenic C4system in a C3crop species.