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Cloning,characterization and expression analysis of a 9-lipoxygenase gene in Gladiolus hybridus
Authors:Qing-long Lian  Hai-bo Xin  Xiong-hui Zhong  Zi-you Zhang  Xiao-xin Li  Xue Yuan  Hao-jun Han  Xiu-li He  Ming-fang Yi
Institution:1. Department of Ornamental Horticulture and Landscape Architecture, China Agriculture University, Yuan Mingyuan Western Road 2#, Beijing 100193, China;2. Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China;3. Agriculture Bureau of Rushan, Rushan 264500, Shandong Province, China
Abstract:Plant lipoxygenases (LOXs) is a functionally diverse class of dioxygenases involved in multiple physiological processes such as growth, senescence, and stress-related responses. Previously, based on the analysis of LOX1 activity and corm formation, it was depicted that LOX1 increased jasmonic acid (JA) synthesis and induced enlargement at top of stolon branches, forming cormels in Gladiolus hybridus. Here, LOX gene (GhLOX1) in full length was isolated from developing corms in G. hybridus. Sequence analysis showed that GhLOX1 was a novel LOX1 gene encoding 9-LOX. Transient expression analysis determined GhLOX1 to be a cytoplasm protein. Real time RT-PCR showed that the mRNA level of GhLOX1 gene correlated positively with LOX activity and was highest in cormels. GhLOX1 mRNA level, LOX activity and endogenous methyl jasmonate (MJ) content in corms increased steadily with a MJ gradient from 0.1 mmol/L to 0.5 mmol/L. Conversely, GhLOX1 mRNA level, LOX activity, and endogenous MJ content in corms steadily decreased with the increasing concentration of Salicylhydroxamic acid (SHAM). At the same time, we observed that MJ and SHAM treatments effected both LOX1 mRNA level and LOX activity, leading to promotion and inhibition of corm formation and enlargement, respectively. These data reveal that GhLOX1 is a novel LOX1 gene encoding a cytoplasm 9-LOX protein and its expression would contribute to LOX activity which plays an important role in corm formation and enlargement. In general, GhLOX1 is an ideal candidate for the promotion of corm formation and enlargement by gene manipulation in G. hybridus.
Keywords:Gladiolus hybridus  9-Lipoxygenases  Salicylhydroxamic acid  Methyl jasmonate  Corm formation
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