Fps Gene Expressed In Tobacco Through Ri-mediated Transformation

fps gene of Mentha spicata L.was cloned and Construction of recombinant clones pBinARfp, And a transgenic system was developed for tobacco via Agrobacterium rhizogenes-mediatcd transformation, Choice 4 lines among resistance to kanamycim(30mg.l^-1). The intergration of fps gene was confirmed PCR and PCR-Southern analysis,and analysis of Northern blot revealed that the foreign fps gene was expressed at the transcriptional level in four hairy root lines; Study to growth of hairy roots and magnify culture in bioreactor show it have large potential; Resistence to fungal with distill substance of hairy roots, which suspension cultures upon addition of fungal elicitor prepared from Alternaria alternata, The result showed obvious resistence to Alternaria alternata and Phytophthora parasitica, It provided approach to utilize plant organize produce biologic pesticide.The main results as follows:1.The study with PBinARfps plasmid transgene wildness Ar1334. Obtain project Ar1334 and transgened tobacco K326 with leaf disc transformation and obtained hairy roots. Design primes with rolC gene in Ri-plasmid T-DNA and PCR, obtain 560bp fragment, proved Ri-plasmid was confirmed; Design primes with fps gene and PCR, obtain 1000bp fragment, proved fps gene was confirmed. fps gene probe was signed with DIG High Prime, PCR- Southern blot analysis with PCR production of fps gene, proved fps gene was expressed in molecule level. Northern blot analysis with RNA of hairy roots, detected objective gene fragment print; which proved fps gene was expressed in transcriptional level. The result proved fps gene was integrated and translated in tobacco genomes and obtained translation fps gene tobacco hairy roots.2. Hairy roots were cultured in MS showed four plants not only have common characteristic, but also have especial in shape. The study to growth trends to hairy root of tobacco in suspendsion cultures. Results showed that translation hairy roots of tobacco showed typical S curve uptrends, Growing period is 30 days, biologic yield accumulate have evidently difference among of single clone plants, the best line-4 fresh weigth achieved 45.6 g/l, the worse line-3 fresh weight achieved 18.9 g/l; line-3 was magnified culture in bioreactor with superior growth condition. The results showed inoculate 2 g/l biologic yield,after a period overgrow bioreactor, biologic yield fresh weight achieved...