The Gene Silence And Function Study Of NADPH Oxidase F In Thelungiella Halophila

The growth, development and propagation of plant can be affected seriously by adversity stress, such as high salt, drought, low temperature and so on. And it's also the important reason for crop reduction. In contrast to O2, these partially reduced or activated derivatives of oxygen [hydroxyl radical (OH·), superoxide anion (O2-), singlet oxygen (1O2) and hydrogen peroxide (H2O2)] are highly toxic and reactive, and can lead to the oxidative destruction of cells. Consequently, the evolution of all aerobic organisms has been dependent on the development of efficient ROS-scavenging. In recent years, a new role for ROS has been identified: the control and regulation of biological processes, such as growth, development, cell cycle, hormone signaling, programmed cell death and biotic and abiotic stress responses. The plant plasma membrane NADPH oxidase has been considered as the primary source of reactive oxygen species such as O2ˉ, H2O2 under the adversity stress. And plasma membrane NADPH oxidase has been paid much more attention to its gene expression, protein structure, membrane topology, and different homologs under various circumstances.In plants, the NADPH oxidase homologs have been named respiratory burst oxidase homologs (Rboh), Rboh gene family contains ten family members (A-J) in Arabidopsis. The ten family members in Arabidopsis are compartmentalized three basic classes on the basis of the tissue specificity: expression throughout the plant (AtRbohD and F), in the roots (AtRboh A-G, I), and in a pollen-specific manner (AtRbohH and J). The conditions of anoxia/hypoxia and nitrogen stress affect the AtRboh family members, and AtRbohC to F are also induced by a variety of biotic stresses. Through analyzing the mutants, we can realize that AtRbohC can react in root hair development, AtRbohD is the major constitutively active form, while AtRbohF exists as a biotic stress-inducible form.Thellungiella halophila which is a relative plant of the Arabidopsis has small size, short life cycle, self-pollination and high seed number, and favorable genetic traits such as self-fertilization, a small genome, efficient transformation and mutagenesis. Therefore Thellungiella halophila become a model plant for studying on the plant salt-tolerant mechanism. In this study, partial fragments of NADPH oxidase F (ThRbohF) gene are obtained from Thellungiella halophila EST library. The silencing vector of gene ThRbohF has been constructed and transformed into Thellungiella halophila by floral dip method. Screening the transgenic seeds by using Basta and have gotten about 10 Basta-resistance plants of the gene silence of ThRbohF, and the 10 Basta-resistance plants have been verified by using semi-quantitative RT-PCR to observe whether the ThRbohF gene expressed in transgenic Thellungiella halophila. This project is aimed to study the function of Thellungiella halophila ThRbohF gene.