Study On The Acquisition And Anti Browning Properties Of Antisense PPO In Agaricus Bisporus

Agaricus bisporus is easy to browning,difficult to store at ordinary temperature after harvest,In this topic,antisense RNA technique was used to inhibit the browning of Agaricus bisporus.Agaricus bisporus PPO inhibitory expression vector and contains strong promoter elements of gpd promoter enhanced expression vector was introduced into As2796 Agaricus bisporus by Agrobacterium tumefaciens-mediated genetic transformation,after the cultivation of Agaricus bisporus experiment,successfully obtained anti browning of Agaricus bisporus storage materials.The main results are as follows:1.Through the study of Agrobacterium tumefaciens to infect different tissue blocks,the efficient transgenic system of Agaricus bisporus mediated by Agrobacterium tumefaciens was established.The mediotrastum are the best ideal material for infection in Agaricus bisporus Agrobacterium transformation,the contamination rate was 10.3%.The transgenic Agaricus bisporus fruiting strains Y1-10 yield reached 15.32 kg/m~2 than the control group increased by41.5%,Z5-5,Y1-10 strains with the excellent properties of the potential that thick of the cap and short sturdy ofthe stem.Z5-19 mushroom cap thickness up to 2.88cm,the single mushroom weight up to 4.36g.2.Analyzed transgenic strain PPO gene expression.by RT-PCR methods detected Agaricus transgenic strains of PPO in expression level and enzyme activity determination,It was found that antisense inhibition of genetically modified Agaricus bisporus PPO gene has a similar expression at the level of transcription and protein expression,there are 65%antisense strains can be inhibited,the inhibits expression level of 50%~75%between.In the enhanced expression of transgenic Agaricus bisporus,the PPO gene was expressed in different degrees of high expression,and 87%of the strains were highly expressed.3.in the study of anti browning of transgenic mushrooms found that enhanced expression of transgenic Agaricus bisporus was significantly better than antisense inhibition of transgenic Agaricus bisporus and CK in the hardness value.which enhanced the expression of strain Z-5-5showed the highest hardness is 4.5N,the CK was 2.9N,Z5-5,Z5-19 and Z6-9 strains in 5 days after storage mushroom cap whiteness value of 80~82.5 is still in the consumer acceptable range,significantly higher than the CK.there are highly correlated between the whiteness and hardness of mushroom cap.The correlation value is as high as 0.73.The resistance to browning and storage of strain Z5-5,Z5-19 and Z6-9 is closely related to the characteristics of high hardness.