title: Induction of reactive oxygen species and the potential role of NADPH oxidase in hyperhydricity of garlic plantlets in vitro.
abstract: Hyperhydricity is a physiological disorder associated with oxidative stress. Reactive oxygen species (ROS) generation in plants is initiated by various enzymatic sources, including plasma membrane-localized nicotinamide adenine dinucleotide phosphate (NADPH) oxidases, cell wall-bound peroxidase (POD), and apoplastic polyamine oxidase (PAO). The origin of the oxidative burst associated with hyperhydricity remains unknown. To investigate the role of NADPH oxidases, POD, and PAO in ROS production and hyperhydricity, exogenous hydrogen peroxide (H2O2) and inhibitors of each ROS-producing enzyme were applied to explore the mechanism of oxidative stress induction in garlic plantlets in vitro. A concentration of 1.5 mM H2O2increased endogenous ROS production and hyperhydricity occurrence and enhanced the activities of NADPH oxidases, POD, and PAO. During the entire treatment period, NADPH oxidase activity increased continuously, whereas POD and PAO activities exhibited a transient increase and subsequently declined. Histochemical and cytochemical visualization demonstrated that specific inhibitors of each enzyme effectively suppressed ROS accumulation. Moreover, superoxide anion generation, H2O2content, and hyperhydric shoot frequency in H2O2-stressed plantlets decreased significantly. The NADPH oxidase inhibitor was the most effective at suppressing superoxide anion production. The results suggested that NADPH oxidases, POD, and PAO were responsible for endogenous ROS induction. NADPH oxidase activation might play a pivotal role in the oxidative burst in garlic plantlets in vitro during hyperhydricity.
authors.affiliation: College of Horticulture, Ministry of Agriculture, Nanjing Agricultural University/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Nanjing, 210095, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Tian
authors.affiliation: College of Horticulture, Ministry of Agriculture, Nanjing Agricultural University/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Nanjing, 210095, People's Republic of China.
authors.forename: Yaqi
authors.initials: Y
authors.lastname: Cheng
authors.affiliation: College of Horticulture, Ministry of Agriculture, Nanjing Agricultural University/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Nanjing, 210095, People's Republic of China.
authors.forename: Xiangyu
authors.initials: X
authors.lastname: Kong
authors.affiliation: College of Horticulture, Ministry of Agriculture, Nanjing Agricultural University/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Nanjing, 210095, People's Republic of China.
authors.forename: Min
authors.initials: M
authors.lastname: Liu
authors.affiliation: College of Horticulture, Ministry of Agriculture, Nanjing Agricultural University/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Nanjing, 210095, People's Republic of China.
authors.forename: Fangling
authors.initials: F
authors.lastname: Jiang
authors.affiliation: College of Horticulture, Ministry of Agriculture, Nanjing Agricultural University/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Nanjing, 210095, People's Republic of China. zpzx@njau.edu.cn.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Wu
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D005737:Garlic / Q000201:enzymology* / Q000502:physiology*; D006861:Hydrogen Peroxide / Q000378:metabolism; D019255:NADPH Oxidases / Q000378:metabolism*; D018384:Oxidative Stress; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000378:metabolism; D010544:Peroxidases / Q000378:metabolism; D018515:Plant Leaves / Q000378:metabolism; D018520:Plant Shoots / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D000097667:Polyamine Oxidase
chemical_list: D006861:Hydrogen Peroxide; D019255:NADPH Oxidases; D000587:Oxidoreductases Acting on CH-NH Group Donors; D010544:Peroxidases; D017382:Reactive Oxygen Species; D013481:Superoxides; D000097667:Polyamine Oxidase
keywords: Apoplast PAO; Cell wall POD; Garlic; Hyperhydricity; NADPH oxidases; Plantlet in vitro; Reactive oxygen species