Faculty of Applied Bioscience, Tokyo University of Agriculture, 1-1-1 Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan.
BMC Plant Biol. 2010 Nov 24;10:261. doi: 10.1186/1471-2229-10-261.
Thellungiella halophila (also known as T. salsuginea) is a model halophyte with a small size, short life cycle, and small genome. Thellungiella genes exhibit a high degree of sequence identity with Arabidopsis genes (90% at the cDNA level). We previously generated a full-length enriched cDNA library of T. halophila from various tissues and from whole plants treated with salinity, chilling, freezing stress, or ABA. We determined the DNA sequences of 20 000 cDNAs at both the 5'- and 3' ends, and identified 9569 distinct genes.
Here, we completely sequenced 1047 Thellungiella full-length cDNAs representing abiotic-stress-related genes, transcription factor genes, and protein phosphatase 2C genes. The predicted coding sequences, 5'-UTRs, and 3'-UTRs were compared with those of orthologous genes from Arabidopsis for length, sequence similarity, and structure. The 5'-UTR sequences of Thellungiella and Arabidopsis orthologs shared a significant level of similarity, although the motifs were rearranged. While examining the stress-related Thellungiella coding sequences, we found a short splicing variant of T. halophila salt overly sensitive 1 (ThSOS1), designated ThSOS1S. ThSOS1S contains the transmembrane domain of ThSOS1 but lacks the C-terminal hydrophilic region. The expression level of ThSOS1S under normal growth conditions was higher than that of ThSOS1. We also compared the expression levels of Na+-transport-system genes between Thellungiella and Arabidopsis by using full-length cDNAs from each species as probes. Several genes that play essential roles in Na+ excretion, compartmentation, and diffusion (SOS1, SOS2, NHX1, and HKT1) were expressed at higher levels in Thellungiella than in Arabidopsis.
The full-length cDNA sequences obtained in this study will be essential for the ongoing annotation of the Thellungiella genome, especially for further improvement of gene prediction. Moreover, they will enable us to find splicing variants such as ThSOS1S (AB562331).
盐芥(Thellungiella halophila,又称盐穗木)是一种模式盐生植物,具有体型小、生命周期短、基因组小等特点。盐芥基因与拟南芥基因具有高度的序列同一性(cDNA 水平为 90%)。我们之前从各种组织和经盐胁迫、冷胁迫、冻胁迫或 ABA 处理的整株植物中生成了盐芥全长 cDNA 富集文库。我们确定了 20000 个 cDNA 的 5' 和 3' 末端的 DNA 序列,鉴定了 9569 个不同的基因。
在此,我们完全测序了 1047 个与非生物胁迫相关基因、转录因子基因和蛋白磷酸酶 2C 基因相关的盐芥全长 cDNA。预测的编码序列、5'UTR 和 3'UTR 与拟南芥的同源基因的长度、序列相似性和结构进行了比较。盐芥和拟南芥同源物的 5'UTR 序列具有显著的相似性,尽管基序发生了重排。在研究与胁迫相关的盐芥编码序列时,我们发现了盐芥盐过度敏感 1(ThSOS1)的一个短剪接变体,命名为 ThSOS1S。ThSOS1S 包含 ThSOS1 的跨膜结构域,但缺乏 C 端亲水区域。在正常生长条件下,ThSOS1S 的表达水平高于 ThSOS1。我们还使用来自每个物种的全长 cDNA 作为探针,比较了盐芥和拟南芥中 Na+转运系统基因的表达水平。在盐芥中,一些在 Na+排泄、区室化和扩散中起重要作用的基因(SOS1、SOS2、NHX1 和 HKT1)的表达水平高于拟南芥。
本研究获得的全长 cDNA 序列对于盐芥基因组的持续注释至关重要,特别是对进一步提高基因预测能力至关重要。此外,它们还使我们能够发现剪接变体,如 ThSOS1S(AB562331)。