Development Of Transgenic Osnar2.1or OSPT6Cotton

Cotton(Gossypium spp.) is an important economy crop, cotton fiber provides the main natural raw materials for the textile industry, its seed is rich in lipid and protein, so cotton has an important strategic status in national economy. However, with the deteriorating global environment, extreme growth environment especially low nitrogen and low phosphorus affect quantity and quality of cotton product, Xinjiang as an example:the cotton needs nitrogen and phosphorus fertilizer about304kg-hm’2and150kg-hm"2respectively for the best cotton nitrogen phosphorus assimilation and dry matter accumulation which restricts the cotton yield and quality improved. Nitrogen and phosphate fertilizer overuse not only increase agricultural production costs, but also a waste of resources and environmental pollution. OsNAR2.1is a accessory proteins of nitrate efficient absorption protein OsNRT cloned from rice, only when OsNAR2.1exists, OsNRT can assimilates the nitrate in the case of low nitrogen, so the OsNAR2.1plays an important role in NO3-assimilation. OsPT6is a major high affinity phosphate transporter gene which has an important role under the phosphorus deficiency condition. So it has an important significance creating nitrogen efficient phosphorus efficient absorption of new cotton material by transgenic methods.In this study, OsNAR2.1plant expression vector transformed into the cotton genome using Agrobacterium-mediated method infect WO receptor hypocotyl. Transgenic plants were analyzed by PCR validation, got35positive clones and70T0generation plants after grafting.Got20positive clones and70T0generation plants transferred OsPT6gene to cotton genome using the same way. All18clones (of61transgenic cotton) T1generation seeds were obtained by the indoor field kanamycin resistance identification and PCR testing, get two pure lines clones; remaining16clones of T1generation after plants selfing. Pure lines of genetically modified cotton get fluorescence quantitative PCR analysis showed OsPT6expressed in transgenic cotton, and the expression is difference between the two clones. Low phosphorus stress screening results showed that the concentration of phosphate in the culture medium as low as0.5mmol/L when cotton began to show significant phosphorus starvation using WO for receptor, therefore,0.5mmol/L as a low-phosphorus stress concentrations, verify OsPT6response in transgenic cotton. Digest the transgenic cotton by heating digestion, using inductively coupled plasma spectroscopy instrument occurs (1CP) to detect phosphorus content of plant digestion solution, the results showed that two gene transfer OsPT6cotton plants were higher in phosphorus element content, clone1was an increase of2.2%compared with the WO group, clone2was4.1%, reaching significant difference. Two clones phosphorus content result coincide with quantitative results indicating OsPT6gene expression promotes the absorption of phosphorus element in cotton.