The disease-resistant transgenic peanut cv ICG 13942 plants were developed by using Tcchitinase-I gene. Agrobacterium tumefaciens strain LBA4404 harboring the binary vector (pBinAR) contains the chitinase ( Tcchitinase-I ) gene and neomycin phosphotransferase resistance ( nptII ) gene. The transformed shoots were developed on selection medium (MMS + 0.5 mg/L IAA + 15 mg/L TDZ + 100 mg/L Kan + 250 mg/L Cefotaxime) from deembryonated cotyledon (DC) explants. Established plantlets were screened for the presence of Tcchitinase-I and nptII genes. Stable integration and expression of the transgenes (T 0 ) were confirmed by using PCR, RT-PCR and Southern blot analyses. The transformation frequency 63.34% was recorded. All the transformed (T 0 ) plants were found normal, flowered and set seeds. After selfing the T 0 plants, a Mendelian inheritance pattern (3:1) for the transgene in T 1 progeny is revealed. T 1 transgenic peanut plants were evaluated for resistance against Cercospora arachidicola, C. personatum and Puccinia arachidis by infection with the microspores using detached leaf assay. These T 1 plants have shown longer incubation, latent period and lower infection frequencies in comparison to non-transformed (WT) plants. The Tcchitinase-I gene expression in resistant transgenic plants was compared to that of a susceptible control. A significant negative correlation was recorded between chitinase activity and the frequency of infection to the three tested disease causing agents.

中文翻译：

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Tcchitinase-I基因在转基因花生（Arachis hypogaea L.）中的表达增强了对叶斑病和锈病的抗性

利用Tcchitinase-I基因开发了抗病转基因花生cv ICG 13942植株。携带二元载体 (pBinAR) 的根癌农杆菌菌株 LBA4404 含有几丁质酶 (Tcchitinase-I) 基因和新霉素磷酸转移酶抗性 (nptII) 基因。在选择培养基（MMS + 0.5 mg/L IAA + 15 mg/L TDZ + 100 mg/L Kan + 250 mg/L 头孢噻肟）上从脱胚子叶 (DC) 外植体培养转化的枝条。筛选建立的小植株中是否存在 Tcchitinase-I 和 nptII 基因。通过使用PCR、RT-PCR和Southern印迹分析证实了转基因(T 0 )的稳定整合和表达。记录的转化频率为 63.34%。发现所有转化的(T 0 )植物均正常、开花并结籽。在自交 T 0 植物后，揭示了 T 1 后代转基因的孟德尔遗传模式 (3:1)。使用离体叶试验通过用小孢子感染来评估T 1 转基因花生植物对花生尾孢、C.personatum和花生锈菌的抗性。与非转化 (WT) 植物相比，这些 T 1 植物显示出更长的孵育时间、潜伏期和更低的感染频率。将抗性转基因植物中的 Tcchitinase-I 基因表达与易感对照的表达进行比较。几丁质酶活性与感染三种测试致病因子的频率之间记录到显着的负相关。使用离体叶测定法通过小孢子感染personatum和Puccinia arachidis。与非转化 (WT) 植物相比，这些 T 1 植物显示出更长的孵育时间、潜伏期和更低的感染频率。将抗性转基因植物中的 Tcchitinase-I 基因表达与易感对照的表达进行比较。几丁质酶活性与感染三种测试致病因子的频率之间记录到显着的负相关。使用离体叶测定法通过小孢子感染personatum和Puccinia arachidis。与非转化 (WT) 植物相比，这些 T 1 植物显示出更长的孵育时间、潜伏期和更低的感染频率。将抗性转基因植物中的 Tcchitinase-I 基因表达与易感对照的表达进行比较。在几丁质酶活性和感染三种测试致病因子的频率之间记录了显着的负相关。