Wang Huabo, Liu Dongcheng, Sun Jiazhu, Zhang Aimin
Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Chaoyang District, Beijing 100101, People's Republic of China.
J Plant Physiol. 2005 Jan;162(1):81-9. doi: 10.1016/j.jplph.2004.07.006.
Differences in gene expression between salinity stressed and normally grown wheat seedlings were compared by the differential display (DD) technique. One DD-derived cDNA clone was characterized as a partial sequence of the wheat asparagine ynthetase (AS) gene by sequence analysis and homology search of GenBank databases. Two AS genes of wheat, TaASN1 and TaASN2, were further isolated by the RT-PCR approach. Comparison of the deduced polypeptide of TaASN1 and TaASN2 with AS proteins from other organisms revealed several homologous regions, in particular, the conserved glutamine binding sites and Class-II Glutamine amidotransferases domain. The functionality of TaASN1 was demonstrated by complementing an Escherichia coli asparagine auxotroph. TaASN1 transcripts were detected in roots, shoots, anthers and young spikes by RT-PCR analysis. Abundance of TaASN1 mRNA in young spikes and anthers was higher than that in shoots and roots under normal growth conditions. TaASN1 was dramatically induced by salinity, osmotic stress and exogenous abscisic acid (ABA) in wheat seedlings. TaASN2 transcripts were very low in all detected tissues and conditions and were only slightly induced by ABA in roots.
采用差异显示(DD)技术比较了盐胁迫下和正常生长的小麦幼苗之间的基因表达差异。通过序列分析和GenBank数据库同源性搜索,将一个来自差异显示的cDNA克隆鉴定为小麦天冬酰胺合成酶(AS)基因的部分序列。通过RT-PCR方法进一步分离出小麦的两个AS基因TaASN1和TaASN2。将TaASN1和TaASN2推导的多肽与其他生物的AS蛋白进行比较,发现了几个同源区域,特别是保守的谷氨酰胺结合位点和II类谷氨酰胺氨基转移酶结构域。通过对大肠杆菌天冬酰胺营养缺陷型进行互补,证明了TaASN1的功能。通过RT-PCR分析在根、茎、花药和幼穗中检测到TaASN1转录本。在正常生长条件下,幼穗和花药中TaASN1 mRNA的丰度高于茎和根。在小麦幼苗中,TaASN1受盐胁迫、渗透胁迫和外源脱落酸(ABA)的显著诱导。TaASN2转录本在所有检测的组织和条件下都非常低,仅在根中受ABA轻微诱导。