León-Galván Fabiola, de Jesús Joaquín-Ramos Ahuizolt, Torres-Pacheco Irineo, Barba de la Rosa Ana P, Guevara-Olvera Lorenzo, González-Chavira Mario M, Ocampo-Velazquez Rosalía V, Rico-García Enrique, Guevara-González Ramón Gerardo
División de Ciencias de la Vida, Departamento de Ingeniería en Alimentos, Campus Irapuato-Salamanca, Universidad de Guanajuato, Ex-Hacienda el Copal km 9, Carretera Irapuato-Silao, CP 36500, Irapuato Guanajuato, Mexico; E-Mail:
Int J Mol Sci. 2011;12(11):7301-13. doi: 10.3390/ijms12117301. Epub 2011 Oct 25.
A germin-like gene (CchGLP) cloned from geminivirus-resistant pepper (Capsicum chinense Jacq. Line BG-3821) was characterized and the enzymatic activity of the expressed protein analyzed. The predicted protein consists of 203 amino acids, similar to other germin-like proteins. A highly conserved cupin domain and typical germin boxes, one of them containing three histidines and one glutamate, are also present in CchGLP. A signal peptide was predicted in the first 18 N-terminal amino acids, as well as one putative N-glycosylation site from residues 44-47. CchGLP was expressed in E. coli and the recombinant protein displayed manganese superoxide dismutase (Mn-SOD) activity. Molecular analysis showed that CchGLP is present in one copy in the C. chinense Jacq. genome and was induced in plants by ethylene (Et) and salicylic acid (SA) but not jasmonic acid (JA) applications in the absence of pathogens. Meanwhile, incompatible interactions with either Pepper golden mosaic virus (PepGMV) or Pepper huasteco yellow vein virus (PHYVV) caused local and systemic CchGLP induction in these geminivirus-resistant plants, but not in a susceptible accession. Compatible interactions with PHYVV, PepGMV and oomycete Phytophthora capsici did not induce CchGLP expression. Thus, these results indicate that CchGLP encodes a Mn-SOD, which is induced in the C. chinense geminivirus-resistant line BG-3821, likely using SA and Et signaling pathways during incompatible interactions with geminiviruses PepGMV and PHYVV.
对从抗双生病毒的辣椒(辣椒属中国辣椒品种BG - 3821)中克隆出的一个类萌发素基因(CchGLP)进行了特性分析,并对表达蛋白的酶活性进行了分析。预测的蛋白质由203个氨基酸组成,与其他类萌发素蛋白相似。CchGLP中还存在一个高度保守的铜锌超氧化物歧化酶结构域和典型的萌发素盒,其中一个包含三个组氨酸和一个谷氨酸。在N端前18个氨基酸中预测有一个信号肽,以及从第44 - 47位残基处有一个推定的N - 糖基化位点。CchGLP在大肠杆菌中表达,重组蛋白表现出锰超氧化物歧化酶(Mn - SOD)活性。分子分析表明,CchGLP在辣椒属中国辣椒品种的基因组中以单拷贝形式存在,在无病原体情况下,乙烯(Et)和水杨酸(SA)处理可诱导其在植物中表达,但茉莉酸(JA)处理则不能。同时,与辣椒金色花叶病毒(PepGMV)或辣椒瓦斯特科黄脉病毒(PHYVV)的不亲和互作会导致这些抗双生病毒植物中CchGLP在局部和系统水平上被诱导,但在一个感病品种中则不会。与PHYVV、PepGMV以及卵菌辣椒疫霉的亲和互作不会诱导CchGLP表达。因此,这些结果表明CchGLP编码一种Mn - SOD,在辣椒属抗双生病毒品系BG - 3821中被诱导,可能在与双生病毒PepGMV和PHYVV的不亲和互作过程中利用SA和Et信号通路。