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The Study On Protoplasts Fusion Between Saururus Chinensis And Houttuynia Cordata

Posted on:2003-06-15Degree:DoctorType:Dissertation
Country:ChinaCandidate:N Q JiangFull Text:PDF
GTID:1100360062486110Subject:Silviculture
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1. The karyotype of Saururus chinensis and Houttuynia cordata. had been analyzed. The results showed that Saururus chinensis' s karyotype formula is 2n=4m+16m+2st, and Houttuynia cordata' s karyotype formula is 2n=14m+10sm02. The Explants of stems,leaves and roots of Saururus chinensis were cultured on MS medium supplemented with 0.2mg/L NAA , l.Omg/L BA and 200mg/l inositol, 0.1%HgCl2 solution as a explants disinfectant and treat by it with in 8 minutes for explants,the frequency of callus induction is better than others.3. There are influences of different carbon resouce, sucrose concentration, phytohormones type, and medium on cell growth of suspension cultures of Saururus chinensis.Among those factors, 30g/L sucrose, 1.5mg/L 2,4-D, l.Omg/L BA and 4.0g DW/L inoculum quantities added in to the MS medium brougt about a increase of absolute cell growth rate.4. The best combination of isolating Protoplasts of Sauru- rus chinensis was the enzyme mixture containing 1.5% Onozuka R-10,0.1%Macerozyme R-10 0.7mol/L mannitol and 10 hours of treating time. The result was obtained by orthogonal design of L9 (34) and L16 (45) . Hou- ttuynia cordata is all the same with Saururus chinens- is except Onozuka R-10. And also studies on factors of influence fision showed that the higher frequency of fu -sion was obtained with 40% polyethylene glycol (PEG) 6000, 15mmol/L CaCl2 2H2O, 0.4mmol/L KH2PO4 and 0.7mol/L mannitol.at pH 9.0 and 1X 106 /ml protop- last density.5. The binuclear heterokaryons frequency reached 7 per cent by polyethylene glycol(PEG) method.Fused protoplasts were cultured in MS90medium containing l.Omg/L NAA , 0.5mg/L BA. First cell division occurred within 5-6 days. 12 weeks later, obtained calli up to 2.0mm in diameter were transferred to MS medium supplemented with l.Omg/L NAA and were cultured for 4 weeks,resulting in rapid callus proliferation. The above 80 per cent hybrid calli have chromosome number from 36 to 50 and some variation in morphology. With increasing generation of subculture, the range of variation of chromosome was much smaller. Chromosome counting of hybrid calli and electron microscope analysis indicated that intergenus somatic hybrid callus was obtained.6. The pharmacologic test result showed that hybrid callus, and the mixture of Saururus chinensis and Houttuynia cordata can increase the thymus weight of mice.
Keywords/Search Tags:Saururus chinensis, Houttuynia cordata, protoplasts fusion, intergenus hybrid callus regeneration
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