Laboratory of Plant Functional Genomics, Center for Genetic Engineering and Biotechnology, PO Box 6162, Havana, 10600, Cuba.
Gene. 2010 Mar 1;452(2):54-62. doi: 10.1016/j.gene.2009.11.011. Epub 2009 Dec 11.
To identify Nicotiana tabacum genes involved in resistance and susceptibility to Rhizoctonia solani, suppression subtractive hybridization was used to generate a cDNA library from transcripts that are differentially expressed during a compatible and incompatible interaction. This allowed the isolation of a protein kinase cDNA that was down-regulated during a compatible and up-regulated during an incompatible interaction. Quantitative RT-PCR analysis of this gene confirmed the differential expression patterns between the compatible and incompatible interactions. Over-expression of this gene in tobacco enhanced the resistance to damping-off produced by an aggressive R. solani strain. Furthermore, silencing of this protein kinase gene reduced the resistance to a non-aggressive R. solani strain. A set of reported tobacco-resistant genes were also evaluated in tobacco plants over-expressing and silencing the protein kinase cDNA. Several genes previously associated with resistance in tobacco, like manganese superoxide dismutase, Hsr203J, chitinases and phenylalanine ammonia-lyase, were up-regulated in tobacco plants over-expressing the protein kinase cDNA. Potentially, the protein kinase gene could be used to engineer resistance to R. solani in tobacco cultivars susceptible to this important pathogen.
为了鉴定与烟草抗根肿病和感病相关的基因，利用抑制性消减杂交技术，从亲和和非亲和互作中差异表达的转录本中构建了一个 cDNA 文库。这使得分离到一个在亲和互作中下调而在非亲和互作中上调的蛋白激酶 cDNA。该基因的定量 RT-PCR 分析证实了亲和和非亲和互作之间的差异表达模式。在烟草中超表达该基因增强了对强致病性根肿菌菌株引起的猝倒病的抗性。此外，沉默该蛋白激酶基因降低了对非致病性根肿菌菌株的抗性。还评估了一组报道的烟草抗性基因在过表达和沉默蛋白激酶 cDNA 的烟草植株中的表达。一些先前与烟草抗性相关的基因，如锰超氧化物歧化酶、Hsr203J、几丁质酶和苯丙氨酸解氨酶，在过表达蛋白激酶 cDNA 的烟草植株中上调。该蛋白激酶基因可能被用于工程改造烟草对这种重要病原体的抗性。
