Studies On High Quality Targeted Pigellia Ternata Genetic Engineering

Pinellia ternate,a species within the genus Pinellia of Araceae,is a kind of important traditional Chinese herb.The dried tuber of Pinellia has the effect of antiemetic,analgesic and sedative.Moreover,the tuber is widely applied to relieve coughing and phlegm.Although P.ternate is widely distributed in China,the germplasm resource becomes limited due to excessive exploitation.The molecular genetic transformation and molecular breeding of P.ternate is also rarely conducted.In this study,to obtain new profile of P.ternata with higher medicinal value,disease-resistance,aseptic seedling of P.ternata originated from Jingzhou(Hubei,China)were got by seedling transformation system.We transferred ER gene,antiapoptotic genes IAP and HMGR gene which are originated from Araidopsis,Spodoptera exigua and Staphylococcus aureus,respectively,were transferred into P.ternata through Agrobacterium-mediated transformation.Meanwhile,BAR gene and beta-glucuronidase gene were also transferred into P.ternate as selection markers.At the sametime,we integrated ERECTA and HMGR genes to a same vector and made the two genes in the same strain for higher expression of both genes.A total of 301 strains were obtained by transferring the five genes into Pinellia by genetic transformation system.PCR analyses showed that 72 clones in these transgenic Pinellia lines appeared positive result.Using Real Time PCR technique for detecting of gene expression in Pinellia,the expression of these genes has been greatly improved compared to wild Pinellia.