Yang Jun, Ji Lianlian, Wang Xingfen, Zhang Yan, Wu Lizhu, Yang Yingna, Ma Zhiying
North China Key Laboratory for Crop Germplasm Resources of Education Ministry, Hebei Agricultural University, Baoding, 071001, China.
Plant Cell Rep. 2015 Aug;34(8):1429-41. doi: 10.1007/s00299-015-1798-5. Epub 2015 May 1.
Expression of DHS1 in cotton is induced upon infection by Verticillium dahliae , and overexpression of GhDHS1 endows transgenic Arabidopsis plants excellent Verticillium resistance. Verticillium wilt is caused by a soil-borne fungus Verticillium dahliae. Resistance in most cotton cultivars is either scarce or unavailable, making Verticillium wilt a major obstacle in cotton production. Here, we identified a 3-deoxy-7-phosphoheptulonate synthase (DHS, EC 4.1.2.15) gene from Gossypium hirsutum, named GhDHS1. Its 1620 bp open reading frame encodes a putative 59.4 kDa protein. Phylogenetic analysis indicated that GhDHS1 is clustered in a clade with potato and tomato DHSs that can be induced by wounding and elicitors, respectively. Expression analysis demonstrated that GhDHS1 is constitutively expressed in cotton roots and stems, but transcripts are rare or non-existent in the leaves. Subcellular localization showed that GhDHS1 occurs in the plastids. When plants of three cultivars were inoculated with V. dahliae, DHS1 expression was more significantly up-regulated in the roots of resistant G. barbadense cv. Pima90-53 and G. hirsutum cv. Jimian20 than in the susceptible G. hirsutum cv. Han208. This suggested that DHS1 is involved in the cotton resistance to Verticillium wilt. Furthermore, GhDHS1 overexpressing transgenic lines of Arabidopsis were developed via Agrobacterium tumefaciens-mediated transformation. Compared with the untransformed WT (wild type), these transgenic plants showed excellent Verticillium wilt resistance with a significantly lower disease index. The overexpressing transgenic lines also had significantly longer primary roots and greatly increased xylem areas under V. dahliae infection. Overall, our results indicate that GhDHS1 performs a role in the cotton resistance to V. dahliae and would be potential to breeding cottons of Verticillium wilt resistance.
棉花中DHS1的表达在大丽轮枝菌感染后被诱导,GhDHS1的过表达赋予转基因拟南芥植株优异的抗大丽轮枝菌能力。黄萎病由土壤传播的大丽轮枝菌引起。大多数棉花品种中抗性稀缺或不存在,使得黄萎病成为棉花生产中的主要障碍。在此,我们从陆地棉中鉴定出一个3-脱氧-7-磷酸庚酮糖合成酶(DHS,EC 4.1.2.15)基因,命名为GhDHS1。其1620 bp的开放阅读框编码一个推定的59.4 kDa蛋白质。系统发育分析表明,GhDHS1与分别可被创伤和激发子诱导的马铃薯和番茄DHSs聚集在一个进化枝中。表达分析表明,GhDHS1在棉花根和茎中组成型表达,但在叶中的转录本稀少或不存在。亚细胞定位显示,GhDHS1存在于质体中。当三个品种的植株接种大丽轮枝菌时,抗性海岛棉品种Pima90-53和陆地棉品种冀棉20的根中DHS1表达上调比感病陆地棉品种邯208更显著。这表明DHS1参与棉花对黄萎病的抗性。此外,通过农杆菌介导的转化培育了拟南芥GhDHS1过表达转基因株系。与未转化的野生型相比,这些转基因植株表现出优异的抗黄萎病能力且病害指数显著更低。在大丽轮枝菌感染下,过表达转基因株系的主根也显著更长且木质部面积大幅增加。总体而言,我们的结果表明,GhDHS1在棉花对大丽轮枝菌的抗性中发挥作用,并且在培育抗黄萎病棉花方面具有潜力。