以铝耐品种和铝敏感品种为材料，以与Ahc-3蛋白（类caspase-3）互作为核心，利用差异转录组学和酵母双杂交等技术，筛选获得不同抗性品种中与Ahc-3蛋白互作的E3泛素连接酶ULE-99和ULE-ZH。利用Nothern blot和West blot检测ULE-99和ULE-ZH在不同铝处理下的转录和翻译水平；利用免疫共沉淀、点突变等试验研究与Ahc-3的相互作用和互作结构域及泛素化水平；构建重组质粒，将基因分别转入烟草野生型及突变体TAhc-3、TAhSAG1、TAhBI-1，检测对Ahc-3、SAG1、AhBI-1等凋亡相关基因表达的影响。通过DNA ladder、TUNLE、线粒体生理等检测野生型和突变型在Al处理前、后细胞程序性死亡（PCD）发生的变化。研究结果对明确不同抗性品种中Ahc-3蛋白泛素化在铝诱导花生根尖PCD中的调控作用与耐铝关系，阐明植物耐铝机制具有重要意义。

This project is to uncover regulation mechanism of caspase-3-like (Ahc-3) protein ubiquitination between different aluminum (Al)-resistant peanut cultivars during Al-induced programmed cell death (PCD) in the root tips of different Al-resistant peanut cultivars. E3 Ubiquitin ligases that interacted with Ahc-3 in Al-resistant and Al-sensitive peanut cultivars, namely ULE-99 and ULE-ZH, respectively, are screened and characterized by using comparative transcriptome analysis and yeast two-hybrid technology. The transcriptional and translational levels of the characterized ULE-99 and ULE-ZH are investigated in corresponding two peanut cultivars. The interaction between ULE-99/ULE-ZH and Ahc-3 and their interacted domains and ubiquitination are detected by using co-Immunoprecipitation, point mutation and in vivo ubiquitination technologies. The recombinant plasmids containing ULE-99 and ULE-ZH, respectively, are constructed and then introduced into wild type tobacco and its mutants TAhc-3, TAhSAG1 and TAhBI-1. The effects on the expression of Ahc-3, SAG1 and AhBI-1 in the constructed transgenic tobacco are observed. PCD is monitored in wild type tobacco and its transgenic plants under Al treatment by DNA ladder, TUNEL and mitochondrial alterations. The conclusion will provide a new sight to understand the regulation mechanism of caspase-3-like protein ubiquitination in PCD and further verify molecular mechanism of Al-resistance in plants.