Zhang Chunzhi, Liu Lei, Wang Xiaoxuan, Vossen Jack, Li Guangcun, Li Tao, Zheng Zheng, Gao Jianchang, Guo Yanmei, Visser Richard G F, Li Junming, Bai Yuling, Du Yongchen
The Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Zhongguancunnandajie 12, 100081, Beijing, People's Republic of China.
Theor Appl Genet. 2014 Jun;127(6):1353-64. doi: 10.1007/s00122-014-2303-1. Epub 2014 Apr 23.
Ph-3 is the first cloned tomato gene for resistance to late blight and encodes a CC-NBS-LRR protein. Late blight, caused by Phytophthora infestans, is one of the most destructive diseases in tomato. The resistance (R) gene Ph-3, derived from Solanum pimpinellifolium L3708, provides resistance to multiple P. infestans isolates and has been widely used in tomato breeding programmes. In our previous study, Ph-3 was mapped into a region harbouring R gene analogues (RGA) at the distal part of long arm of chromosome 9. To further narrow down the Ph-3 interval, more recombinants were identified using the flanking markers G2-4 and M8-2, which defined the Ph-3 gene to a 26 kb region according to the Heinz1706 reference genome. To clone the Ph-3 gene, a bacterial artificial chromosome (BAC) library was constructed using L3708 and one BAC clone B25E21 containing the Ph-3 region was identified. The sequence of the BAC clone B25E21 showed that only one RGA was present in the target region. A subsequent complementation analysis demonstrated that this RGA, encoding a CC-NBS-LRR protein, was able to complement the susceptible phenotype in cultivar Moneymaker. Thus this RGA was considered the Ph-3 gene. The predicted Ph-3 protein shares high amino acid identity with the chromosome-9-derived potato resistance proteins against P. infestans (Rpi proteins).
Ph-3是首个克隆的番茄抗晚疫病基因,编码一种CC-NBS-LRR蛋白。由致病疫霉引起的晚疫病是番茄最具毁灭性的病害之一。源自醋栗番茄L3708的抗性(R)基因Ph-3,对多种致病疫霉分离株具有抗性,已广泛应用于番茄育种计划。在我们之前的研究中,Ph-3被定位到9号染色体长臂远端含有R基因类似物(RGA)的区域。为了进一步缩小Ph-3区间,使用侧翼标记G2-4和M8-2鉴定出更多重组体,根据Heinz1706参考基因组,将Ph-3基因定位到一个26 kb的区域。为了克隆Ph-3基因,利用L3708构建了一个细菌人工染色体(BAC)文库,并鉴定出一个包含Ph-3区域的BAC克隆B25E21。BAC克隆B25E21的序列显示,目标区域仅存在一个RGA。随后的互补分析表明,这个编码CC-NBS-LRR蛋白的RGA能够互补品种Money maker的感病表型。因此,这个RGA被认为是Ph-3基因。预测的Ph-3蛋白与9号染色体来源的马铃薯抗致病疫霉蛋白(Rpi蛋白)具有高度的氨基酸同一性。