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长心卡帕藻原生质体分离、培养研究
张思, 刘翠, 金月梅, 池姗, 唐贤明, 陈傅晓, 方诩, 刘涛
2014, 33(10): 114-123. doi: 10.1007/s13131-014-0546-y
关键词: 长心卡帕藻, 原生质体, 再生植株, 类愈伤组织, 再生新枝
以长心卡帕藻为材料,利用海螺酶和鲍酶两种海藻工具酶与纤维素酶成功制备其原生质体。确定了最适酶比例为20%鲍酶与12%纤维素酶,渗透剂为2.0 mol/L葡萄糖,最适酶解条件为30℃黑暗酶解4.0 h,原生质体密度为32.60×104 个/mL,产量为65.20×104 个/g。20℃,光强1500-2000 lx,光周期12 h/d培养,原生质体出现类愈伤组织细胞团和再生植株两种分化途径,酶解后组织块培养出现了再生新枝和类愈伤组织细胞团两种分化途径。
Genetic transformation of marine Actinomycete sp. isolate M048 and expression of a recombinant plasmid carrying the apc gene
HOUYanhua, LIFuchao, QINSong, WANGQuanfu
2006(6): 145-152.
关键词: allophycocyanin, protoplast, intergenericconjugantion, exconjugant, marineActinomycetes
Optimal conditions for protoplasts formation of marine Actinomycete sp.isolate M048 were described, dense and disperse mycelia were cultured in SGGP medium, 0.5% glycine, lysozyme exposure (2 mg/cm3, 37℃, 40 min), and the concentration of sucrose in protoplast buffer was 0.4 mol/dm3 for keeping the balance of osmotic pressure.Using PEG-mediated protoplasts transformation, the transformation frequency was 89 transformants per microgramme of pIJ702.Meanwhile, an effective transformation procedure was established based on intergeneric conjugation from E.coli ET12567 (pUZ8002) using shuttle vectors pPM801, pPM803 and a (ψ)C31-derived integration vector pIJ8600 containing ori T and att P fragments.Transformation frequencies were 5.30×10-4±0.26×10-4, 8.92×10-4±0.19×10-4 and 6.38×10-5±0.41×10-5, respectively.Further, the heterologous expression of the allophycocyanin gene (apc) in the strain M048 was used to demonstrate this transformation system.SDS-PAGE and Western blot analysis confirmed the expression of recombinant APC (rAPC).
Taxonomy and ultrastructure of five species of Tetraselmis (Prasinophyceae) isolated from China seas
ZhangChengwu, HuHongjun
2002(4): 557-579.
关键词: Taxonomy, ultrastructure, Tetraselmis, pyrenoid
Comparative ultrastructural investigations on twenty-three isolates of Tetraselmis from the South China Sea, East China Sea and Huanghai Sea, have revealed that these isolates belong to:(1) Tetraselmis chui Butcher; (2) T. cordiformis (H. J. Carter) Stein; (3) T. helgolandica Kylin; (4) T. suecica (Kylin) Butcher; (5) T. guangdongensis sp. nov. Except T. helgolandica,the others are new records in China. T. guangdongensis sp. nov. is a new species. Its external and anatomical features closely resemble those of T. impellucida McLachlan et Parke. As in that species the pyrenoid is penetrated from many directions by cytoplasmic channels delimited by a double membrane.The protoplast withdraws from the apical portion of the theca, that portion which overlaid the trough inverts and pops out in the form of a teat. But unlike that species a starch sheath is present; pyrenoid matrix is surrounded by thylakoids which intervenes between the matrix and the starch sheath surrounding pyrenoid; the theca is stratified.