【作者】 江年琼；

【导师】 谢碧霞；

【作者基本信息】 中南林学院 ， 森林培育学， 2002， 博士

【摘要】 本研究的主要内容及结果有以下几点： 1对三白草科的三白草（Saururus chinensis （Lour.）Baill.）和鱼腥草（Houttuynia cordata Thunb）的核型进行了分析，结果表明：三白草和鱼腥草的核型公式分别为K（2n）=22=4m+16sm+2st和K（2n）=24=14m+10sm。 2三白草的茎、叶和地下根茎均可诱导出愈伤组织，但以嫩茎段作为外植体，以MS为基本培养基，添加NAA0.2mg／L和BA1.0mg／L及肌醇200mg／L时愈伤组织诱导率较高。外植体消毒以0.1％HgCl2处理8min污染率最低。 3在三白草悬浮培养时，不同碳源、蔗糖浓度、激素种类、不同培养基和接种量等对三白草悬浮培养细胞的生长均有影响。其中以3％蔗糖作为碳源，以MS培养基中附加2，4-D1.5mg／L和BA1.0mg／L为好，接种量为4.0gDW／L时，细胞的绝对生长速率较快。 4通过L9（3⁴）和L16（4⁵）的正交设计，对酶溶液最佳配方及影响原生质体融合的因素进行了研究，结果表明：分离三白草原生质体酶溶液的组成以1.5％的纤维素酶、0.1％果胶酶、0.7mol／L甘露醇和10h的酶解时间为好。分离鱼腥草原生质体的酶溶液除纤维素酶的浓度为1.0％外，其余与三白草相同。由40％PEG（6000）、15mmol／L CaCl₂ 2H₂O、0.4mmol／L KH₂PO₄和0.7mol／L甘露醇组成的、pH为9.0的融合剂效果较好。原生质体密度为1×10⁶个／ml较佳。 5用PEG诱导融合的双核异核体率为11％左右，将融合原生质体培养在含有NAA1.0mg／L和BA0.5mg／L的MS培养基中，5～6d后，再生细胞发生首次分裂，培养12周后，将直径约2.0mm的愈伤组织转移到添加NAA1.0mg／L的MS培养基上培养4周，愈伤组织迅速增殖。杂种愈伤组织中，80％以上的染 三自草和鱼腥草原生质体融合的研究 色体数目在36～50条之间，但形态学上有一定的变异。随着继代时间的延长， 染色体的变异系数越来越小。研究结果表明，三白草和鱼腥草原生质体融合获得 了属间杂种愈伤组织。 6对杂种愈伤组织和两种原植物的混合物，分别用醇和水进行提取，再对提 取物进行药理实验，结果表明：杂种愈伤组织和两原植物混合后的水提取液可使 小臼鼠胸腺的重量增加，p＜O．05，但两者间无显著性差异，p＞0刀5。更多还原

【Abstract】 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.更多还原