Production And Salt-tolerance Analysis Of Transgenic Potato With Arabidopsis AtNHX1 Gene Driven By Different Promoters

Soil salinization is the golobel problem in agriculture,soil salization has serious harm in growing plant.it is the new important way to breed new variety by biology technology.Potato is the fourth food crop in the world ,and it is also important stuff of machining material of vegetable and food.But it is sensitive into salization,and salization is bad for its growing.so it has developed for breeding new crop varieties with strong salt tolerance and good synthetical characteristics potato veriety has important singnification biology technology.This research used rd29A promoter which driving by Arabidopsis vacuole membrane Na+/H+ antiporter gene(AtNHX1) to transfer AtNHX1 making potato genetic transformation,and identified NaCl intimidation of eight series lines which having transferred the AtNHX1driven by CaMV 35S promoter into microtubers,now ,the research result as follow:1.Optimization of potato's genetic transformation system: (1)making microtuber slice of potato as transformation material,certain microtuber slice differentiates fitting medium was: MS+1.00mg/L IAA+0.20mg/L GA3+2.00mg/L ZT+0.50mg/L 6-BA;(2)The Kanamycin selective concentration is made sure as 50mg/L in differentiation phases.2.The transgenic plants of potato were obtained: The transgenic plants which rooted in the Kanamycin medium were examined by distinctive PCR,indication that AtNHX1 gene already transformed into the genome of potato; Analyzing salt-tolerance further on the different NaCl concentration culture medium,we can see that the transgenic plant could grow regulary in 0.6% NaCl medium but the un-trangenic plant couldn't grow regularly.3.Whether the eight series lines of transferred CaMV35S+AtNHX1 gene or un-transgenic plants were harmed under high concentration NaCl (0.6%和0.9%)stressed , all showed that the basal old leaves becoming yellow and dead first by prolong of the time;later ,the leaves in middle will curl,withered,die,but the leaves which near to growing point are tiny to be yellow,and they showed curl,stopping growing or growing slowly,but the harm of transgenic is obvious lower than un-transgenic plant.Un-transgenic plant all died under 0.9% NaCl stress,but transgenic plant only showed that most of the basal old leaves died,and most of the middle leaves curled,partly series lines showed withered,and that resulted transgenic plant and untransgenic plant had obvious difference between the produce capability,single potato weight and the biggest potato weight.4.With the increase of NaCl concentration and prolonging of stress time,the contents of Na+ in leaves of the untrasgenic plant was more distinct than the transgenic plant,especially under 0.9%NaCl stressed after 40 days, the contents of Na+ was 2.0-6.2 times compared with the transgenic plant in the same desposition,therefore,the un-transgenic plant died under this concentration in the end .5.Because the proliferation of the height of plants under NaCl limitaion can't reflectted the growing situation between the transgenic plant and the un-transgenic plant,so this characteristic is not fitting to estimate the resistance ability.Extent of the leaf areas of the un-transgenic plant stopped nearly under 0.9% NaCl stressed long time,and the old leaves of un-transgenic plant were plentiful died,new leaves stopped growing,and died in the end, because the transgenic plant were transmitted into effective salt-tolerance gene which can reduce the harm of Na+,so the plant still showed preferable growth. Thereby,0.9% NaCl is maybe the deadly concentration of the microtuber plant on Gannongshu 2.6.With the increase of NaCl concentration and the stress time,Chlorophyll contents and membrance perability of the transgenic plant and the un-transgenic plant all showed drop or increase trend,and the range in drop or increase of the un-transgenic plant were higher than the transgenic plant,therefore,we can see the difference of leaves'color under high concentration of NaCl for a long time by naked eyes.7.With the increase of NaCl concentration,the proline content of un-transgenic plant increased largely,here,plant growth was affected obviously.But the proline contents of transgenic plant was droped more than the un-transgenic plant,so the magnitude of this value shows the degree that the plant were harmed.8.General experiment evaluation,the transgenic plant series lines 8,18and19 have better ability resistance the NaCl stress.