Identification of new gene expression regulators specifically expressed during plant seed maturation
Identification of new gene expression regulators specifically expressed during plant seed maturation
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DOI：
10.1093/jxb/erj138
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发表时间：
2006-06-01
影响因子：
6.9
通讯作者：
Van Wuytswinkel, Olivier
中科院分区：
文献类型：
--
作者：
Gutierrez, Laurent;Conejero, Genevieve;Van Wuytswinkel, Olivier
采用cDNA - AFLP方法对亚麻（Linum usitatissimum）进行研究，以鉴定在种子成熟过程中特异性表达的基因。在产生的20000个cDNA - AFLP标签中，有486个因其在成熟过程中的种子特异性表达而被选中。与公开可用的数据库进行比对时，其中一半与已知的植物序列呈现出显著的相似性。所获得的结果证实了该方法的准确性，因为在此筛选中鉴定出了许多先前被描述为仅在植物种子中表达的基因。重点关注与在转录、转录后或翻译后水平参与基因表达调控的植物调节因子相似的序列。通过实时逆转录聚合酶链反应（RT - PCR）方法，证实了其中13种调节因子的种子特异性表达动力学，这些调节因子此前从未被鉴定在种子成熟过程中表达。其中，通过原位杂交方法表明，一个非LEC1样HAP3家族的亚麻基因和一个亚麻MYB因子在亚麻胚胎的特定组织中表达。通过对这些亚麻基因及其拟南芥对应基因的表达动力学比较，发现新的HAP3基因应与一种普遍存在的种子成熟机制相关，而一个新的MYB因子似乎与一种更具种子特异性的成熟机制相关。这些结果表明，亚麻数据库不仅在鉴定种子成熟过程中表达的新基因方面有用，而且能够突出保守和非保守种子成熟机制之间的区别。
A cDNA-AFLP approach on Linum usitatissimum (flax) was used to identify genes specifically expressed during the seed maturation process. Among the 20000 cDNA-AFLP tags produced, 486 were selected for their seed-specific expression during maturation. When compared with the publicly available databases, half of them presented some significant similarity with known plant sequences. The results obtained confirmed the accuracy of the approach as numerous genes previously described as being expressed exclusively in plant seeds were identified in this screen. The focus was on sequences similar to plant regulators involved in the control of gene expression, either at the transcriptional, post-transcriptional, or post-translational levels. Using a real-time RT-PCR approach, seed-specific expression kinetics were confirmed for 13 of these regulators that were never characterized for being expressed during seed maturation. Among these, a flax gene of the non-LEC1-like HAP3 family and a flax MYB factor were shown to be expressed in specialized tissues of flax embryo using an in situ hybridization approach. By expression kinetic comparison between these flax genes and their Arabidopsis counterparts, it was found that the new HAP3 gene should be related to a ubiquitous seed maturation mechanism, while a new MYB factor appears to be related to a more seed-specific maturation mechanism. These results demonstrate the utility of the flax database in not only identifying new genes expressed during seed maturation but also in being able to highlight the distinction between conserved and non-conserved seed maturation mechanisms.