As an important intracellular signaling module, the mitogen-activated protein kinase (MAPK) cascades have been previously implicated in signal transduction during plants responsing to various environmental stresses as well as pathogen attack. The mitogen-activated protein kinase kinase acts as the convergent point of MAPK cascades during a variety of stress signaling. In this study, a novel MAPKK gene, ZmMKK1, in maize (Zea mays L.) belonging to group A MAPKK was isolated and functionally characterized. ZmMKK1 was mainly localized in the cytoplasm and its constitutive kinase-active form ZmMKK1DD was localized in both cytoplasm and nucleus. QRT-PCR analysis uncovered that ZmMKK1 expression was triggered by abiotic and biotic stresses and exogenous signaling molecules. Moreover, hydrogen peroxide (H2O2) and Ca(2+) mediated 12°C-induced up-regulated expressing of ZmMKK1 at mRNA level. Ectopic expression of ZmMKK1 in tobacco (Nicotiana tabacum) conferred tolerance to chilling stress by higher antioxidant enzyme activities, more accumulation of osmoregulatory substances and more significantly up-expression of ROS-related and stress-responsive genes compared with empty vector control plants. Furthermore, ZmMKK1 played differential functions in biotrophic versus necrotrophic pathogen-induced responses. These results suggested ZmMKK1 played a crucial role in chilling stress and pathogen defense in plants.
Keywords: 3,3-diaminobenzidine; ABA; APX; CAT; CaMV; Chilling stress; DAB; DMTU; ETH; GFP; H(2)O(2); MAPK; MDA; MS; MeJA; Murashige-Skoog; NBT; O(2)(−); OE; ORF; POD; Pathogen defense; REL; ROS; Reactive oxygen species; SA; SAR; SOD; Vec plant; WT; ZmMKK1; ZmMKK1-overexpressing; abscisic acid; ascorbate peroxidase; catalase; cauliflower mosaic virus; dimethylthiourea; empty vector control plant; ethephon; green fluorescent protein; hydrogen peroxide; malondialdehyde; methyl jasmonate; mitogen-activated protein kinase; nitroblue tetrazolium; open reading frame; peroxidase; qRT-PCR; quantitative real time-polymerase chain reaction; reactive oxygen species; relative electrolyte leakage; salicylic acid; superoxide anion radical; superoxide dismutase; systemic acquired resistance; wild-type.
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