Howarth Jonathan R, Parmar Saroj, Barraclough Peter B, Hawkesford Malcolm J
Plant Science Department, Rothamsted Research, West Common, Harpenden, Hertfordshire, AL5 2JQ, UK.
Plant Biotechnol J. 2009 Feb;7(2):200-9. doi: 10.1111/j.1467-7652.2008.00391.x.
A sulphate deficiency-induced gene, sdi1, has been identified by cDNA-amplified fragment length polymorphism (AFLP) analysis utilizing field-grown, nutrient-deficient wheat (Triticum aestivum var. Hereward). The expression of sdi1 was specifically induced in leaf and root tissues in response to sulphate deficiency, but was not induced by nitrogen, phosphorus, potassium or magnesium deficiency. Expression was also shown to increase in plant tissues as the external sulphate concentration in hydroponically grown plants was reduced from 1.0 to 0.0 mm. On this basis, sdi1 gene expression has potential as a sensitive indicator of sulphur nutritional status in wheat. Genome-walking techniques were used to clone the 2.7-kb region upstream of sdi1 from genomic DNA, revealing several cis-element motifs previously identified as being associated with sulphur responses in plants. The Arabidopsis thaliana gene most highly homologous to sdi1 is At5g48850, which was also demonstrated to be induced by sulphur deficiency, an observation confirmed by the analysis of microarray data available in the public domain. The expression of Atsdi1 was induced more rapidly than previously characterized sulphur-responsive genes in the period immediately following the transfer of plants to sulphur-deficient medium. Atsdi1 T-DNA 'knockout' mutants were shown to maintain higher tissue sulphate concentrations than wild-type plants under sulphur-limiting conditions, indicating a role in the utilization of stored sulphate under sulphur-deficient conditions. The structural features of the sdi1 gene and its application in the genetic determination of the sulphur nutritional status of wheat crops are discussed.
利用田间种植的缺素小麦(普通小麦品种Hereward),通过cDNA扩增片段长度多态性(AFLP)分析鉴定出一个硫酸盐缺乏诱导基因sdi1。sdi1的表达在叶片和根系组织中因硫酸盐缺乏而特异性诱导,但不受氮、磷、钾或镁缺乏的诱导。随着水培植物外部硫酸盐浓度从1.0 mM降至0.0 mM,植物组织中的表达也显示增加。在此基础上,sdi1基因表达有潜力作为小麦硫营养状况的敏感指标。采用基因组步移技术从基因组DNA中克隆sdi1上游2.7 kb区域,揭示了几个先前鉴定为与植物硫反应相关的顺式元件基序。与sdi1同源性最高的拟南芥基因是At5g48850,经公共领域的微阵列数据分析证实,该基因也受硫缺乏诱导。在植物转移到缺硫培养基后的一段时间内,Atsdi1的表达比先前表征的硫反应基因诱导得更快。Atsdi1 T-DNA“敲除”突变体在硫限制条件下比野生型植物维持更高的组织硫酸盐浓度,表明在缺硫条件下储存硫酸盐的利用中发挥作用。本文讨论了sdi1基因的结构特征及其在小麦作物硫营养状况遗传测定中的应用。