Echenique V., Stamova B., Wolters P., Lazo G., Carollo L., Dubcovsky J.
Department of Agronomy and Range Science, University of California, One Shields Avenue, Davis CA 95616-8515, USA.
Theor Appl Genet. 2002 Apr;104(5):840-844. doi: 10.1007/s00122-001-0849-1. Epub 2002 Feb 8.
The frequency of Ty1- copia-type and Ty3- gypsy-type retrotransposons in the International Triticeae EST Consortium (ITEC) database (61,942 sequences: 82% wheat, 10% barley, 8% rye) and the DuPont EST database (86,628 wheat sequences) was estimated using BLASTN searches. These ESTs were obtained from 94 cDNA libraries from different tissues (leaves, roots, spikes, flowers and seeds) and different growing conditions, excluding subtracted and normalized cDNA libraries. Triticeae EST databases were screened using four different Ty1- -copia-type, 12 reverse transcriptase sequences, and three Ty3- gypsy-type Triticeae retrotransposon sequences. Using a selection threshold of BLASTN scores higher than 100 or E values smaller than e(-20), 0.145% of the ESTs were found to be significantly similar to at least one of the retrotransposons used in the search (0.064% Ty1- copia, 0.081% Ty3- gypsy). This percentage increased to 0.176% when the BLASTN threshold was changed to E<e(-10). The percentage of ESTs similar to retrotransposons was significantly higher ( P < 0.05) in cDNA libraries from leaf tissues than in cDNA libraries from roots, anthers, or spikes. In addition, the percentage of ESTs similar to retrotransposons in cDNA libraries from plants under stress conditions (0.25% at E<e(-20), and 0.30% at E<e(-10)) was three to four folds higher ( P < 0.0001) than in cDNA libraries from plants grown under normal conditions (0.07% at E<e(-20), and 0.09% at E<e(-10)). Identification of retrotransposons within the Triticeae EST databases provides an indirect estimation of the patterns of transcriptional activity of these repetitive elements and is important to improve the annotation of genomic sequences used to search these EST databases.
利用BLASTN搜索估计了国际小麦族EST联盟(ITEC)数据库(61942条序列:82%为小麦,10%为大麦,8%为黑麦)和杜邦EST数据库(86628条小麦序列)中Ty1-copia型和Ty3-gypsy型逆转座子的频率。这些EST来自94个不同组织(叶、根、穗、花和种子)以及不同生长条件下的cDNA文库,不包括扣除和标准化的cDNA文库。使用四个不同的Ty1-copia型、12个逆转录酶序列以及三个Ty3-gypsy型小麦族逆转座子序列对小麦族EST数据库进行筛选。使用BLASTN得分高于100或E值小于e(-20)的选择阈值时,发现0.145%的EST与搜索中使用的至少一种逆转座子有显著相似性(0.064%为Ty1-copia型,0.081%为Ty3-gypsy型)。当BLASTN阈值改为E<e(-10)时,这一百分比增加到0.176%。与逆转座子相似的EST在叶组织cDNA文库中的百分比显著高于根、花药或穗的cDNA文库(P<0.05)。此外,胁迫条件下植物的cDNA文库中与逆转座子相似的EST百分比(E<e(-20)时为0.25%,E<e(-10)时为0.30%)比正常生长条件下植物的cDNA文库高3至4倍(P<0.0001)(E<e(-20)时为0.07%,E<e(-10)时为0.09%)。在小麦族EST数据库中鉴定逆转座子可间接估计这些重复元件的转录活性模式,对于改进用于搜索这些EST数据库的基因组序列注释非常重要。