来源于耐旱荒漠植物蛋白CkND对棉花黄萎病的抑制作用及其抗旱性研究

 利用蛋白组学策略分离了荒漠植物-牛心朴子（Cynanchum komarovii）抗病、抗旱功能相关蛋白CkND。以棉花黄萎病菌为供试菌株，研究了蛋白CkND对棉花黄萎病的抑制作用。结果表明：蛋白CkND对棉花黄萎病菌菌丝生长及孢子萌发具有较好的抑制作用。离体试验中，100 mg·L^-1的蛋白CkND对棉花黄萎病菌菌丝生长抑制效果在24 h、48 h、72 h分别为79.53%、83.01%、87.50％，35 mg·L^-1的蛋白CkND对孢子萌发抑制率为100%；活体试验中，蛋白CkND对棉花黄萎病具有显著的防治效果。另外，本研究将CkND基因构建到pCAMBIA1304表达载体CaMV35S启动子下游，将pCAMBIA1304-CkND通过农杆菌介导转化法转入野生型拟南芥植株中。将33个株系的转pCAMBIA1304 -CkND的 T₂代拟南芥和21株转pCAMBIA1304的T₂代拟南芥，同时进行抗旱性试验。结果显示，转pCAMBIA1304 的T₂代拟南芥14个株系枯死，而转pCAMBIA1304-CkND的 T₂代拟南芥有24个株系的转基因植株存活，且叶片保持绿色，植株生长良好，根系、须根发达，须根数目明显增多。转pCAMBIA1304-CkND拟南芥株高增加了30%，显著提高了植物抗旱性。

Studies on the Inhibition of Plant Derived Protein CkND Against Verticillium dahliae and Its Drought Resistance

Protein CkND, a novel disease resistant and drought-resistant protein, was extracted form the roots of Cynanchum Komarovii by method of proteomics. In this study, we tested the fungicidal activities and resistance of protein CkND against Verticillium dahliae. The results showed that short protein CkND may have a good inhibition on the spore germination and mycelial growth of Verticillium dahliae. In vitro test suggested that 100 mg·L^-1 of protein CkND on mycelial growth at 24 h, 48 h, 72 h was 79.53%, 83.01% and 87.50％， respectively ，and 35 mg·L^-1of protein CkND on spore germination of Verticillium dahliae was 100%. In vivo test showed that protein CkND had good inhibitory effects against Verticillium dahliae. In addition, the CkND gene was constructed into the downstream of the CaMV35S promoter of the pCAMBIA1304 expression vector and transfer of pCAMBIA1304-CkND was transferred into Arabidopsis thaliana mediated by Agrobacterium tumefaciens. We take 33 pCAMBIA1304-CkND T₂ transgenic plants and 21 pCAMBIA1304 T₂ transgenic plants into drought-resistant experiments at the same time. The results showed that 14 of the pCAMBIA1304 T₂ transgenic plants died, but 24 pCAMBIA1304-CkND T₂ transgenic plants survived. The leaves kept green, the plant growed well, roots and fibrous roots developed strongly, and the number of fibrous roots increased obviously. The drought-resistant of pCAMBIA1304-CkND transgenic plants increased 30%, which obviously enhance the drought-tolerance of plants.