Qin Juan, Huang Chuanming, He Fuxin, Zhu Xiaoguo, Zhang Yang, Kang Zhensheng, Guo Jun
Wei Sheng Wu Xue Bao. 2014 Nov 4;54(11):1296-303.
To clone calcium-dependent protein kinase gene (camk) from Puccinia striiformis f. sp. tritici (Pst) and analyze its function.
The cDNA full-length of Pscamk was isolated by using reverse transcriptional-PCR (RT-PCR), and gene expression profile at different morphological stages was analyzed via quantitative real-time--PCR (qRT-PCR). Pst urediospores were treated with CaMK suppressor KN-93 and germination rate was investigated.
A gene cDNA full-length with 1 620 bp was obtained and designated as Pscamk. qRT-PCR analysis showed Pscamk expression was highly induced in the early stages of Pst infection and reached the maximum at 6 h post inoculation (hpi) as 20.74-fold as that in the control (0 hpi). With increasing of the concentration of CaMK suppressor KN-93, germination rate of Pst urediospores was gradually decreased. The germination rate was reduced to 8.02%, only 12% of the control, under 1.4 μmol/L KN-93 treatment at 10 h after incubation at 9 degrees C.
Pscamk might play a role in germination and germ tube elongation of Pst urediospores. This study provides a basis for exploring pathogenesis of calcium signaling pathway during Pst infection.
从条锈菌中克隆钙依赖蛋白激酶基因（camk）并分析其功能。
采用逆转录聚合酶链反应（RT-PCR）分离条锈菌钙依赖蛋白激酶基因（Pscamk）的cDNA全长，并通过实时荧光定量PCR（qRT-PCR）分析其在不同形态阶段的基因表达谱。用CaMK抑制剂KN-93处理条锈菌夏孢子，检测其萌发率。
获得一个1620bp的基因cDNA全长，命名为Pscamk。qRT-PCR分析表明，Pscamk在条锈菌侵染早期高表达，接种后6h达到最高，是对照（0h）的20.74倍。随着CaMK抑制剂KN-93浓度的增加，条锈菌夏孢子萌发率逐渐降低。在9℃培养10h后，1.4μmol/L KN-93处理下，夏孢子萌发率降至8.02%，仅为对照的12%。
Pscamk可能在条锈菌夏孢子萌发和芽管伸长过程中发挥作用。本研究为探索条锈菌侵染过程中钙信号途径的致病机制提供了依据。
