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生长素/吲哚乙酸(Aux/IAA)基因家族在裸子植物火炬松(Pinus taeda)中是保守的。

Aux/IAA gene family is conserved in the gymnosperm, loblolly pine (Pinus taeda).

作者信息

Goldfarb Barry, Lanz-Garcia Carmen, Lian Zhigang, Whetten Ross

机构信息

Department of Forestry, North Carolina State University, Raleigh, NC 27695-8002, USA.

出版信息

Tree Physiol. 2003 Dec;23(17):1181-92. doi: 10.1093/treephys/23.17.1181.

Abstract

We isolated five members of the Aux/IAA gene family in loblolly pine (Pinus taeda L.). Degenerate primers complementary to conserved regions of angiosperm Aux/IAA genes were used to amplify fragments that were, in turn, used as probes to screen a cDNA library constructed from auxin-treated hypocotyls. The five unique clones, named PTIAA1-5, contain the four highly conserved domains that are characteristic of the Aux/IAA proteins. All clones contain the bipartite nuclear localization signal (NLS) between Domains I and II that is predicted in most angiosperm Aux/IAA genes, but only one, PTIAA2, contains the conserved NLS in Domain IV. The five invariant residues in Domain II that have been found to constitute part of a protein destabilization element in Arabidopsis thaliana (L.) Heynh. are conserved in all the PTIAAs. A postulated phosphorylation site located between Domains I and II and proximal to the conserved bipartite NLS was conserved in 20 out of 36 genes in this analysis, including the pine genes. Transcripts of all five PTIAAs accumulated specifically in the hypocotyls in response to exogenous auxin treatment and were induced by all auxins tested. Transcript abundance above basal levels in response to 1-naphthaleneacetic acid treatment was first detected after 10 min (PTIAA3) to 3 h (PTIAA2) in the different genes and remained above basal levels throughout 7 days. Induction of PTIAA2 was inhibited by the protein synthesis inhibitor cycloheximide, indicating that PTIAA2 is a secondary response gene. Phylogenetic analysis showed that all five pine genes clustered within a single class (Class I) of the dendrogram. Clone PTIAA2 has a sequence that is relatively distinct from the other four and is the most closely related to the angiosperm genes of Class I. Class I contains both primary and secondary auxin response genes, suggesting that it is the original lineage and that other gene classes have evolved subsequent to the angiosperm/gymnosperm divergence.

摘要

我们从火炬松(Pinus taeda L.)中分离出了生长素/吲哚乙酸(Aux/IAA)基因家族的五个成员。与被子植物Aux/IAA基因保守区域互补的简并引物被用于扩增片段,这些片段随后被用作探针来筛选由生长素处理过的下胚轴构建的cDNA文库。这五个独特的克隆,命名为PTIAA1 - 5,包含Aux/IAA蛋白特有的四个高度保守结构域。所有克隆在结构域I和II之间都含有双分型核定位信号(NLS),这在大多数被子植物Aux/IAA基因中都有预测,但只有一个,即PTIAA2,在结构域IV中含有保守的NLS。在拟南芥(Arabidopsis thaliana (L.) Heynh.)中已发现结构域II中的五个不变残基构成了蛋白质不稳定元件的一部分,在所有的PTIAA中这些残基都是保守的。在该分析中,位于结构域I和II之间且靠近保守双分型NLS的一个假定磷酸化位点在36个基因中的20个中是保守的,包括松树基因。所有五个PTIAA的转录本在对外源生长素处理的响应中在下胚轴中特异性积累,并且被所有测试的生长素诱导。在不同基因中,对1 - 萘乙酸处理的响应中,转录本丰度高于基础水平在10分钟(PTIAA3)到3小时(PTIAA2)后首次被检测到,并在7天内一直高于基础水平。PTIAA2的诱导被蛋白质合成抑制剂环己酰亚胺抑制,表明PTIAA2是一个次级响应基因。系统发育分析表明,所有五个松树基因都聚集在树状图的一个单一类别(I类)中。克隆PTIAA2的序列与其他四个相对不同,并且与I类被子植物基因关系最密切。I类包含初级和次级生长素响应基因,这表明它是原始谱系,并且其他基因类别是在被子植物/裸子植物分化之后进化而来的。

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