Lilley Catherine J, Urwin Peter E, Johnston Katherine A, Atkinson Howard J
Centre for Plant Sciences, University of Leeds, Leeds, LS2 9JT, UK.
Plant Biotechnol J. 2004 Jan;2(1):3-12. doi: 10.1046/j.1467-7652.2003.00037.x.
The expression patterns of three promoters preferentially active in the roots of Arabidopsis thaliana have been investigated in transgenic potato plants in response to plant parasitic nematode infection. Promoter regions from the three genes, TUB-1, ARSK1 and RPL16A were linked to the GUS reporter gene and histochemical staining was used to localize expression in potato roots in response to infection with both the potato cyst nematode, Globodera pallida and the root-knot nematode, Meloidogyne incognita. All three promoters directed GUS expression chiefly in root tissue and were strongly up-regulated in the galls induced by feeding M. incognita. Less activity was associated with the syncytial feeding cells of the cyst nematode, although the ARSK1 promoter was highly active in the syncytia of G. pallida infecting soil grown plants. Transgenic potato lines that expressed the cystatin OcIDeltaD86 under the control of the three promoters were evaluated for resistance against Globodera sp. in a field trial and against M. incognita in containment. Resistance to Globodera of 70 +/- 4% was achieved with the best line using the ARSK1 promoter with no associated yield penalty. The highest level of partial resistance achieved against M. incognita was 67 +/- 9% using the TUB-1 promoter. In both cases this was comparable to the level of resistance achieved using the constitutive cauliflower mosaic virus 35S (CaMV35S) promoter. The results establish the potential for limiting transgene expression in crop plants whilst maintaining efficacy of the nematode defence.
为了研究拟南芥中优先在根部活跃的三种启动子在转基因马铃薯植株中对植物寄生线虫感染的响应模式,对来自TUB-1、ARSK1和RPL16A这三个基因的启动子区域与GUS报告基因进行了连接,并利用组织化学染色来定位马铃薯根中对马铃薯胞囊线虫(Globodera pallida)和根结线虫(Meloidogyne incognita)感染的表达情况。所有这三种启动子主要在根组织中指导GUS表达,并且在南方根结线虫取食诱导形成的虫瘿中强烈上调。虽然ARSK1启动子在感染土壤栽培植株的马铃薯胞囊线虫的多核巨细胞中高度活跃,但与胞囊线虫的多核巨细胞相关的活性较低。对在这三种启动子控制下表达半胱氨酸蛋白酶抑制剂OcIDeltaD86的转基因马铃薯株系进行了评估,在田间试验中评估其对马铃薯胞囊线虫属(Globodera sp.)的抗性,在隔离条件下评估其对南方根结线虫的抗性。使用ARSK1启动子的最佳株系对马铃薯胞囊线虫的抗性达到70±4%,且没有相关的产量损失。使用TUB-1启动子对南方根结线虫实现的最高水平的部分抗性为67±9%。在这两种情况下,这与使用组成型花椰菜花叶病毒35S(CaMV35S)启动子实现的抗性水平相当。这些结果确立了在限制作物中转基因表达的同时维持对线虫防御功效的潜力。