The Cloning And Sequence Analysis Of NADPH Oxidase Gene In Heave

"Tap Panel Dryness (TPD)", a serious physiological disease in Hevea, is still a matter of debate as its complicate causality. It may be produced spontaneously or be induced through various kinds of internal and external factors.As the application of modem molecular biology and gene engineering techniques on researching the physiology and biochemistry in Hevea, the isolation of Hevein genes (hevl), and the purification and location of NADPH oxidase on the membrane of lutoids, the mechanism and molec μ lar reg μ lation of " Tap Panel Dryness " are brought to light. The mechanism of Tap Panel Drynes is summed up that: When NADPH oxidase on the member of lutoids is activated by strong stimulation of ethylene or over exploitation, NADPH oxidase is activated to release active oxygen abundantly, the active oxygen can destroy the cell membrane by oxidizing the lipoid in the member of lutoids, which cause the lutoids be broken, hevein be releasen from lutoids. As a result the latex' is consolidated in-situ, or nucleic acid and proteins are induced to be disorder, which cause rubber tree fail to produce latex and form "Tap Panel Dryness". During theformation of "Tap Panel Dryness", NADPH oxidase is a key oxidase that it is activated by the ethylene in vitro and the enzyme activity is increased with elevated Ca2+ levels in vitro. However, how NADPH oxidase in the member of lutoids is activated by strong stimulation of ethylene and over exploitation, the understanding is still unclear. Thus, it is extremely important to clone the gene of NADPH oxidase in the member of lutoids. In this research, we utilized the method of homologous clone with SMART RACE technology (rapid amplification of cDNA ends technology) to clone this gene. According to the conserved region of NADPH oxidase genel to synthesize 3'RACE-GSP for 3'RACE reaction, then design 5'RACE-GSP for 5'RACE reaction on basis of 3'RACE reserved region. After spliced the overlaps between 5' and 3' race product, we obtained a 2178bp sequence, which contain a complete open reading frame from 141bp to 1742bp. The analysis of sequence alignment showed no homologous regions are matched toknown genes, thus the gene is a new gene in Hevea. Originally, the homology of NADPH oxidase gene is low among plant, microbe and...