Genetic Transformation Of Barley By HgNHX1 In Halogeton Glomeratus And Stress Analysis Of Transgenic Barley

Barley(Hordeum vulgare L.)is a gramineous plant,mainly used in the manufacture of beer and feed.It has characteristics of drought resistance,salt tolerance and so on.As the global climate and ecological environment degradation continues,drought and high salinity are two major environmental factors that limit the growth of barley.Cultivating barley varieties of high drought and salt tolerance by using genetic engineering become the urgent requirements of barley cultivation.The recipient material is the basis of genetic transformation.It is very important to find a system with wide variety of sources,convenient preparation,limited small by genotype and season and high frequency regeneration.In this study,Na~+/H~+ antiporters(NHX)Hg NHX1 gene from Halogeton glomeratus was introduced into barley by Agrobacterium-mediated method.The transgenic positive plants were obtained.And the resistance of transgenic barley T1 plants was analyzed.The main findings are as follows:One: The factors influencing the high frequency regeneration system of barley stem were studied by using the common cultivar Ganpi 3,Ganpi 4 and Ganpi 6 as the materials.The best way of de-husking was to use 30% sulfate and shake for an hour in 30℃and 180 r/min.The best method of surface sterilization was 75% ethanol for 1.5 min and 2% sodium hypochlorite for 15 min.The length of basal segments was 2 mm and the seedling age was 3 d.3 mg/L 2,4-D was favorable for callus induction,and the regeneration ability of Ganpi 4 was stronger.Two: The genetic transformation of barley by Hg NHX1 gene from Halogeton glomeratus,using Agrobacterium-mediated method,vacuum osmotic treatment 5 min,ultrasonic treatment for 30 s,and 100 r/min shaking 30 min,instantaneous infection was higher.23 resistant plants were obtained.By PCR molecular detection,7 positive plants were found.And T1 generation seeds were obtained.Three: The stress resistance analysis of T1 transgenic barley showed that the expression of Hg NHX1 gene in barley increased under drought and salt stress,the salt resistance of transgenic Hg NHX1 barley was not obvious,but drought resistance increased.