【作者】 姜新兵；

【导师】 陈力耕；

【作者基本信息】 浙江大学 ， 果树学， 2004， 硕士

【摘要】 本论文首次对香榧组织培养进行研究，分别对香榧愈伤组织的诱导，丛生芽增殖、腋芽萌发和体细胞胚诱导等方面进行研究。并通过体细胞胚的途径首次建立了香榧完整的再生系统，这为以后开展香榧的遗传转化和品种改良研究奠定了基础。主要研究结果如下： 1 不同外植体，其愈伤组织的诱导率和继代能力不同。叶片、茎段、芽、假种皮、胚的诱导率可分别达到79.1％、80.6％、77.1％、65.9％、69.8％。其中由叶片、茎段、芽、假种皮诱导的愈伤组织不能继续继代，均褐化死亡。只有由胚形成的愈伤组织可以在SH+6-BA0.1-0.5mg／l+2,4-D0.1-0.5mg／L或SH+6-BA0.1-0.5mg／l+NAA0.5-1.0mg／L的培养基上继续继代。 2 在胚愈伤组织的继代过程中，通过添加1％的活性炭可以有效的防止香榧胚愈伤组织的褐化问题，并且对于愈伤组织的继代培养和增殖都具有很好的效果 3 利用腋芽增殖和丛生芽诱导的方法均未能获得香榧的完整的再生植株苗。 4 继代形成的胚性愈伤组织在1／2 SH+BA 0.1 mg／L+NAA 0.1 mg／L的培养基上体细胞胚诱导效果最佳，其诱导率可达70.3％，诱导的体细胞胚在合适的培养基上可以继续继代和增殖，其中以BA 0.5 mg／L+NAA 0.5 mg／L且黑暗培养的增殖率最高，增殖倍数达3.0，且体细胞胚的生长势最好。 5 继代的体细胞胚在培养基为1／2 SH或1／2 SH+BA 0.1 mg／L+NAA 0.1 mg／L体细胞胚均能萌发，且能够形成完整植株。更多还原

【Abstract】 In this research study, tissue culture of Torreya grandis. was investigated which includes callus induction, adventitious shoot regeneration and somatic embryogenesis. Plantlets were obtained for the first time through somatic embryogenesis.Various explants have different callus induction rate. From leaf, stem, bud , aril and immature embryo 79.1 % 80.6% 77.1 % 68.9% and 69.8 % callus was induced. Callus from leaf, stem and aril was not stable and couldn’t proliferate on subsequent subcultures due to browning which leads to its death. The callus from embryo was sub-cultured on SH media supplemented with 0.1-0.5mg/L-1 6-BA and 0.1-0.5mg/L-12,4-D or the same media with 0.1-0.5mg/L-1 6-BA and 0.5-l.0mg/ L-1 NAA.It was observed that activated carbon not only effectively control the browning problem but also have a good effect on the callus subculture and proliferation. With the addition of 0.1% activated carbon prevented calli browning.After several subcultures of the callus, it was changed in to embryogenic state on 1/2 SH supplemented with BA 0.1 mg/L-1 + NAA 0.1 mg/L-1. High frequency (70.3%). somatic embryogenesis was achieved and embryoids can keep on growing and proliferating on appropriate medium. The embryoids grew well with high proliferation frequency (3.00) on 1/2 SH supplemented with BA 0.5 mg/L-1 + NAA 0.5 mg/L-1 in the dark condition.Somatic embryos were germinated and the plantlets were obtained on 1/2 SH media supplemented with 0.1 mg/L-1BA +0.1 mg/L L-1 NAA. This is the first report on tissue culture and plant regeneration through somatic embryogenesis and will establish a base for the genetic transformation and varietal improvement of Torreya grandis in the future.更多还原