非洲菊胚性愈伤组织悬浮培养方法建立及其诱变条件研究

以紫罗兰绿心、洋红黑心、绿心黄瓣、鹅黄黑心的花托和花梗，白色黑心的叶片为外植体，在pH值为5．8、蔗糖30g／L和琼脂8g／L的条件下诱导愈伤组织，筛选出的最佳愈伤组织诱导培养基配方是MS＋6B—A1．0mg／L＋NAA0．2mg／L＋2，4-D1．0mg／L诱导出的愈伤转到MS＋6-BA2．0mg／L＋NAA0．2mg／L。适合非洲菊固体和液体培养基上愈伤增长的配方是MS＋6-BA2．0mg／L＋NAA0．2mg／L。20Cy的Co^60辐射剂量和0．1％（V／V）的EMS浓度是较适合的诱变剂量．可以作为半致死剂量进行下一步大批量愈伤组织的诱变处理，从中筛选耐寒或花色特异等突变体。

铈对马铃薯悬浮培养的愈伤组织中花青苷积累及其生物合成基因表达的影响

马铃薯中的花青苷主要为锦蔡素和天竺蔡素的衍生物。花青苷的产出率与生物量和花青苷的含量密切相关。本研究表明，0．1mmol·L^-1Ce^4＋能促进细胞和愈伤组织的生长，而1mmol·L^-1Ce^4＋抑制生长，2mmol·L^-1Ce^4＋对细胞有害而导致细胞死亡。不同浓度牟Ce^4＋能诱导正常的细胞和愈伤组织变红并积累花青苷。用0．1mmol·L^-1Ce^4＋处理15d的细胞和愈伤组织花青苷的含量最高，其次是1mmol·L^-1Ce^4＋处理18d的细胞和愈伤组织花青苷的含量，而最低浓度（0．01mmol·L^-1Ce^4＋）和最高浓度（2mmol·L^-1Ce^4＋）不利于花青苷的产生和积累。半定量RT-PCR分析表明，用0．1mmol·L^-1Ce^4＋处理悬浮培养的愈伤组织能显著诱导花青苷生物合成途径中五个基因（CHS，F3H，F3′5′H，DFR和3GT）的表达，其表达量与花青量的含量呈正相关。

Optimization of Inducing Conditions for Loose Callus of Pinellia ternata

[Objective] The research aimed to establish the cell suspension culture system of Pinellia ternate.[Method] The petiole of Pinellia ternate was as the explant.The orthogonal test was used to study the influences of four plant growth substances（2,4-D,NAA,picloram and KT）and their mixture ratios on the formation of loose callus.[Result] The induction effect of 2,4-D and picloram on the petiole callus of Pinellia ternate was the most significant.Then,the second ones were KT and NAA.The optimal medium which induced Pinellia ternate petiole to form the loose callus was MS＋0.5 mg/L of 2,4-D＋1.0 mg/L of NAA＋1.0 mg/L of picloram＋1.5 mg/L of KT.[Conclusion] The research laid the foundation for extracting the active ingredient from the cell suspension of Pinellia ternate and producing the artificial seed.

Callus Induction and Cell Suspension Culture of Eggplant(Solanum melongena L.)

[Objective] The aim of this article is to establish a cell suspension culture system for eggplant. [Method] Using orthogonal design, appropriate media were screened for callus induction, subculture and cell suspension culture. [Result] The appropriate medium for callus induction was MS supplemented with 0.2 mg/L 6-BA and 0.2 mg/L NAA; the appropriate subculture medium was MS supplemented with0.2 mg/L 6-BA, 0.2 mg/L NAA, and 0.1 mg/L KT; the suitable medium for cell suspension culture was liquid MS medium supplemented with 0.4 mg/L NAA and 0.2mg/L. Cell growth curve was similar to ＂S＂ shape in the above suspension medium.The cell growth included four phases, the initial phase（1-3 d）, the logarithmic phase（3-7 d）, the steady phase（7-8 d）, and the decline phase（8-11 d）. With the increasing culture time, the number of suspension cells increased, and it reached the maximum value at the 7thd, about 3.8 ×105cells/ml. Then the number of cells began to decline rapidly. The cell vigor was the highest at the initial phase. Suspension cells grew best in the liquid MS medium supplemented with NAA（0.4 mg/L）and 6-BA（0.2 mg/L）. The mitotic index reached the maximum, about 4.1% in the above medium, which suggested that this medium was suitable for cell suspension culture of eggplant. [Conclusion] Cell suspension culture system of eggplant provides a significant method for eggplant biotechnology. Genetic transformation and mutants screening can be carried out with this system.

Study on the Browning in Cell Suspension Culture of Taxus cuspidata

[Objective] This study aimed to investigate the browning of T. cuspidata cells in suspension culture and provide the guidance for the cell suspension culture of T. cuspidata. [Method] T. cuspidata callus was used as experimental materials, to explore the effect of different medium, N/P ratio, pH, shaking speed, illumination time and light intensity and other factors on browning of T. cuspidata cells in suspension culture. [Result] Non-browning callus was transferred to 2MB5 medium （pH 7.0） for illumination culture at 22℃ under light intensity of 1 500 lx with shaking speed of 90 r/min for 24 h. Results showed that the cell browning was significantly inhibited. [Conclusion] This study laid the foundation for cell suspension culture of T. cuspidata and had important significance to the large-scale industrial production of paclitaxel.

Optimization of Cell Line Building for Taxus chinensis var. mairei

[Objective] The paper aims to optimize the abduction and domestication conditions of Taxus chinensis var.Mairei callus.[Method] We compared the efficiencies of callus between different explants and investigated the multiplication conditions of callus and suspended cell culture conditions with the buds,young stems and young leaves from T.chinensis var as the explants.[Results] The effect was the best with the bud as the explants; the best way for sterilizing the explants of T.chinensis var mairei was：streptomycin detergent for 2 h ＋ suds for 3 h ＋ 75% alcohol disinfection for 30 s ＋ 10% sodium hypochlorite solution for 25 min ＋ 1‰ mercury chloride for 10 min; the optimum formula of callus subculture was：B5 ＋ 4.0 mg/L NAA ＋ 0.5 mg/L 2,4-D ＋ 0.2 mg/L GA ＋ 0.5 mg/L 6-BA ＋ 2 g/L AC.[Conclusion] This research built the high efficient regeneration system of T.chinensis var.

Somatic embryogenesis in wild relatives of cotton (Gossypium Spp.)

Wild cotton species can contribute a valuable gene pool for agronomically desirable cultivated tetraploid cultivars. In order to exploit diploid cotton a regeneration system is required to achieve transformation based goals. The present studies aimed at optimizing the conditions for regeneration of local varieties as well as wild species of cotton. Different callus induction media were tested with varying concentrations of hormones in which sucrose was used as nutritional source. Different explants (hypo-cotyls, cotyledon, root) were used to check the regeneration of both local cotton plants and wild relatives using T & G medium, BAP medium, CIM medium, EMMS medium, and cell suspension medium. Different stages of embryogenicity such as early torpedo stage, late torpedo stage, heart stage, globular stage and cotyledonary stage were observed in wild relatives of cotton. The results of this study pave the way for establishing future transformation methods.