玉米耐旱自交系在干旱条件下的mRNA差异表达

用16%PEG-6000对玉米耐旱自交系“81565”进行模拟干旱处理,用DD-PCR技术研究干旱与正常浇水对照的mRNA差异表达,发现3个干旱条件下差异表达的特异片段MD1、MD2和MD3。MD1和MD2在干旱条件下减量表达,MD3为干旱诱导表达。序列分析和同源性比对表明,MD1与玉米叶绿体基因组中编码参与RNA转录本Ⅱ型内含子剪切的成熟酶的matK基因有93%的相似性,MD2与极端耐旱的Sporobolus stapfianus的丝氨酸/苏氨酸2C型蛋白磷酸酶基因PP2C的相似性达99%,MD3与属天冬氨酸特异性半胱氨酸蛋白酶类的水稻metacaspase基因有99%的相似性。根据各自序列相似同源基因的功能推测,MD1、MD2和MD3三个片段可能与“81565”的耐旱机制有关。     

Induced cell death in Ceratocystis fimbriata by pro-apoptotic activity of a natural organic compound,perillaldehyde, through Ca2+ overload and accumulation of reactive oxygen species

Black rot caused by the fungus Ceratocystis fimbriata causes notable losses in sweet potato production. Perillaldehyde (PAE), a secondary metabolite in perilla, was studied to determine its antifungal effects on mycelial growth and spore germination of C. fimbriata. The effects of PAE on cell wall integrity and cell membrane permeability were also investigated. To elucidate the possible mechanisms of cell death triggered by PAE, sensitivity of C. fimbriata to PAE toxicity was determined by cytoplasmic and mitochondrial calcium ion concentrations ([Ca²⁺]c and [Ca²⁺]m), reactive oxygen species (ROS), mitochondrial membrane potential (MMP), cytochrome c (cyt c) release, metacaspase activation, phosphatidylserine (PS) externalization and DNA fragmentation. The results suggest that mycelial growth and spore germination were inhibited by PAE in a dose-dependent manner. Ceratocystis fimbriata spores treated with PAE experienced dramatic Ca²⁺ overload and elevated ROS production. Compared to untreated controls, the proportion of fluorescent cells stained with the ROS indicator DCFH-DA and treated with a range of PAE concentrations from 0.0625 to 0.50 μL mL⁻¹ increased by 2.9 ± 0.79% to 27.1 ± 0.38%. Ca²⁺ overload and ROS accumulation induced depolarization of the MMP, contributing to mitochondrial dysfunction. Cyt c was released from the mitochondria to the cytosol, triggering metacaspase activation. The significant antifungal activity of PAE on C. fimbriata was demonstrated by these studies, suggesting that PAE has the potential for wide application to postharvest management of tuber crops, in addition to the application to above-ground fruit and vegetables that have been previously investigated.     

稻瘟病菌（Magnaporthe oryzae）的凋亡诱导和检测

 使用低浓度H2O2和高浓度NaCl诱导稻瘟病菌的凋亡,检测了产生的凋亡特征。结果显示：诱导后稻瘟病菌具有典型的凋亡特征,如胞内活性氧累积、磷脂酰丝氨酸从细胞膜的内表面转移到外表面、染色体浓缩和DNA断裂。进一步研究发现：凋亡关键性酶类caspase的抑制剂Z-VAD-FMK能够显著影响稻瘟病菌的孢子萌发和附着胞形成过程。典型的稻瘟病菌孢子由3个细胞组成;在Z-VAD-FMK的作用下,2个以上细胞同时萌发产生芽管的比例由34.1％增加到66.0％,同时形成附着胞的源细胞也以顶端细胞为主转变为以基部细胞为主。这些结果说明凋亡参与了稻瘟病菌的孢子萌发和附着胞形成过程。