The Immunological Characteristics Of Rubiscos From Different Ecotypes Of Reeds In The Desert Region In Hexi Corridor

Phragmites communis Trin. is a kind of herbage plant living for several years. The representative environment is freshwater or salt lake and marshland. For adapting to the different environments, the shape of the reeds living in the marshland of Hexi corridor has changed very much, but they are still belonged to a same species. According to the characteristics of the living environments, the reeds are divided into four different ecotypes. For a long period of time, our laboratory was engaged in the studies of the different ecotypes reeds from the sides of plant physiology, biochemistry and molecular biology. In this paper, we analysed the immunological characteristics of Rubisco and the large and small subunits from two kinds of ecotypes reeds with the immunological methods.Ribulose-1, 5-Bisphosphate Carboxylase was purified from swamp-reeds and dune-reeds by 35% to 55% ammonium sulfate fractionation, DEAE-Sepharose C1-6B ion exchange and Sephacryl S-300 HR Chromatography. SDS-PAGE results showed that there were only Rubisco large and small subunit bands and one Rubisco subunit binding band. Then Rubisco large and small subunits were separated by SDS-PAGE. Polyclonal antibodies against Rubisco complete enzyme, large and small subunits were generated in male mice. Western blotting analysis showed:A. Swamp-reeds Rubisco complete enzyme antiserum could react with Rubisco complete enzyme from swamp-reeds, dune-reeds. The immunological reactivity of swamp-reeds Rubisco complete enzyme antiserum with swamp-reeds was stronger than dune-reeds. Swamp-reeds Rubisco complete enzyme antiserum could react with Rubisco large subunits of swamp-reeds, dune-reeds and spinaches, but not the small subunits of all. The immunological reactivity of swamp-reeds Rubisco complete enzyme antiserum with dune-reeds was stronger than spinaches, but lighter than swamp-reeds.B. Swamp-reeds and dune-reeds Rubisco large subunits antiserum could recognize the Rubisco complete enzyme of swamp-reeds, dune-reeds, and there was no remarkable difference between the immunological reactivity of swamp-reeds and dune-reeds. Swamp-reeds and dune-reeds Rubisco large subunits antiserum could recognize the large subunits of swamp-reeds, dune-reeds and spinaches, but not small subunits of all. The immunological reactivity of the antiserum with spinaches was lighter than swamp-reeds and dune-reeds.C. We could not detect the antibodies of Rubisco small subunits from swamp-reeds and dune-reeds with the same preparation methods as Rubisco large subunits.D. Spinach Rubisco complete enzyme antiserum could recognize the Rubisco large subunits of spinaches, swamp-reeds, dune-reeds and rice. It could only react with its own small subunits. The immunological reactivity of spinaches Rubisco large subunits antiserum with rice was stronger than sweep-reeds and dune-reeds, and lighter than spinaches.So these results shows, there are same or similar immunological groups between theRubisco complete enzyme from swamp-reeds and dune-reeds, and these immunological groups mainly lies in the Rubisco large subunits, and there is part of consistency on immunological characteristics. The immunological differences of Rubisco complete enzyme and the large subunits between swamp-reeds and dune-reeds are very small, and the differences are large between the reeds and spinach. The Rubisco small subunits of reeds have no antigenicity or its antigenicity is light, so it is difficult to prepare the small subunit antibody. Spinach Rubisco complete enzyme antiserum can only recognize its own small subunits, but not the small subunits of swamp-reeds, dune-reeds and rice. So there are large immunological differences between spinaches Rubisco and reeds Rubisco, and the differences also lies between the monocotyledons and dicotyledonous.