hrpZ_(Psta)在转基因大豆中定量表达与疫霉根腐病和灰斑病抗性相关研究

Gene expression and disease resistance of transformed hrp_(ZPsta)gene in transgenic soybean

以T5转hrpZPsta基因大豆JN29-705-15和JL30-187为材料,利用实时荧光定量PCR技术(Quantitative Real Time PCR,qRT-PCR)检测了目标基因在转基因大豆不同组织中的表达量,分别利用下胚轴侵染法和叶面喷施法鉴定疫霉根腐病抗性和灰斑病抗性,并分析目的基因表达量与疫霉根腐病和灰斑病抗性的相关性。结果表明:hrpZPsta基因在大豆的叶、茎、根、籽粒中均有表达,二个株系平均相对表达量分别为8.2/6.1、0.9/0.7、6.5/4.6和0.8/0.7;T5转hrpZPsta基因大豆抗疫霉根腐病和灰斑病能力与野生型相比均有所提高,JN29-705-15对疫霉根腐病抗性从感病提高到中抗,而JL30-187从中抗提高到抗病;hrpZPsta基因在叶中的表达量也与抗灰斑病能力呈正相关,与病情级别呈极显著负相关。试验结果初步证明了外源基因hrp ZPsta在大豆植株中的表达量与受体植株对疫霉根腐病和灰斑病抗性存在一定的相关性。

T5generation of transgenic soybean- lines JN29- 705- 15 and JL30- 187 with hrp ZPstagene were investigated by real- time quantitative polymerase chain reaction technique( qRT- PCR) to analysis the relationship between expression of the target gene in different tissues of the transgenic soybean and their resistance to Phytophthora root rot and gray leaf spot. The results showed hrp ZPstagene was expressed in soybean leaves,stems,roots and seeds,the average relative expression levels of the two strains were 8. 2 /6. 1,0. 9 /0. 7,6. 5 /4. 6 and 0. 8 /0. 7,respectively; the resistance to Phytophthora root rot and gray leaf spot disease were improved compared with wild- type; compared to control,the resistance against Phytophthora root rot disease of JN29- 705- 15 increased from sensitive to medium- resistant; and resistance of JL30- 187 increased from medium to full- resistant; the expression levels of hrp ZPstagene in leaves was positively related to resistance of gray leaf spot,and significant negative correlation to the disease levels. These results indicated that the expression level of hrp ZPstawas correlated to resistance to both Phyophthora root rot and gray leaf spot in transgenic soybean.