The leaves of Saussurea involucrate were used as experiment materials and were induced embryogenic callus,then using them to isolate protoplast.The influential factors of plasmolysis time,concentration of osmotic solution,enzyme combinations and digestion time on protoplast isolation from embryogenic callus were studied.It is suitable to indicate embryogenic calluses when the murashige and skoog medium which was MS+2,4-D 0.1 mg/L+BA 0.5 mg/L+NAA 1.5 mg/L+glucose 30 g/L.When the embryogenic calluses were used to isolation,the optimal condition was enzyme solution of 2.5% Cellulase Onzuka R-10,0.2% Mecerozyme R-10,1.0% Pectinase,0.1%MES and 10%Mannitol for digesting 14～16 h at pH 5.8 and the temperature of(26±1)℃.

Genetic diversity of transgenic wheat with different phenotypes,‘spring wheat 761’and Leymus racemosus was analyzed by random amplified polymorphic DNA(RAPD)with 160 arbitrary oligonucleotide primers.Five primers with good repetition and high polymorphism were screened.There were several special bands the same as the donor in the transgenic wheat.The genetic similarity coefficient and clustering analysis also confirmed that the gene fragments of L.racemosus had been integrated into the genome of transgenic wheat.

We used somatic embryogenesis to regenerate snow lotus (Saussurea involucrata Kar.et Kir.). For induction of calli, cotyledons were cultured on solid MS medium with 0.5 mg.L-1 2,4-D and 0.05-1 mg.L-1 BA. Globe-shaped yellowish-green calli were chosen from the first subculturing for suspension culture in liquid MS medium containing 0.05-0.1 mg.L-1 2,4-D. After 20 days of culture, many globular embryoids were produced. Polyethylene glycol and gibberellic acid (GA) were added during subculture for differentiation and growth of somatic embryos. Plantlets were regenerated when somatic embryos were transferred to MS medium with 5 mg.L-1 GA3.

BNYVV CP and BBSV CP gene were transformed into sugarbeet of diferent variety using agrobacterium tumfaciens-mediated method.From kanamycin resistance regenerate plant,DNA fregment were obtained by PCR detection,transgenic plant were preliminarily proved.

从新疆维吾尔族健康人造血干细胞中提取总RNA,用反转录(reverse transcription,RT)和降落PCR(touchdoWn PCR,TD-PCR)相结合的方法扩增人细菌透性增加蛋白(human bactericidal/permeability-increasing protein,hBPI)的cDNA,将其克隆到pEGFP-N1载体上并进行DNA序列测定.结果表明,所克隆的hBPIcDNA全长为1,464个碱基,其序列与GenBank中另外4个序列进行了比对,有两个碱基与其它4个序列不同:其中第576位碱基其它序列为G,该位置碱基是C;第676位碱基其它序列为A,该位置碱基是G.其中第676位碱基的变化导致第185位氨基酸由Lys改变为Glu.

The paper dealt with the study on the peroxidase and esterase isozymes as well as change of soluble protein content in the process of direct shoots organogenesis and genetic transformation of sugerbeet in vitro by polyarclamide gel electrophoresis(PAGE).The results showed that peroxidase(POD),isoesterase(EST)isoenzyme and soluble protein content appeared the changes of enzyme zymogram number and colour shade by adding benzylaminopurine(6-BA)to the medium,which related with increase of metabolizing activity.And after genetic transformation,bands of the peroxidase and esterase isozymes and soluble protein came forth obviously colour darken and number increase,which do farther research to identify the resons that resistance to stress was enhanced or natural transgene.

The Saussurea involucrate embryogenic induction culture(MS+2,4-D 0.5 mg/L+6-BA 0.05 mg/L) was made,including somatic cell embryogenesis induction culture(MS+2,4-D 0.1 mg/L),somatic cell embryo differentiation culture(MS+PEG 25 g/L),somatic cell embryo germination(MS+GA3 5 mg/L) and lustihood seedling culture(MS+NAA 0.1 mg/l+6-BA 0.3 mg/L)and the plantlet regeneration was realized through somatic cell embryogenesis way,with rooting rate of 100% and survival rate of 60%,which provided a base for manual culture of Saussurea involucrate cosmically.

目的:探讨人细菌透性增加蛋白(hBPI)在哺乳细胞中的表达和亚细胞定位。方法:以本人克隆得到的质粒pBE1为模板,利用一对带有Kozak序列以及删除终止密码子的引物进行PCR,获得的产物与pEGFP-N1载体连接,构建pBE2哺乳细胞特异表达载体并稳定转染MCF-7细胞,获得了转基因细胞系。提取其基因组DNA,PCR扩增检查BPIcDNA片段和EGFP片段是否已整合入MCF-7细胞基因组中。提取总RNA,通过RT-PCR方法检查BPI-EGFP在转录水平的表达。Western blot进一步鉴定融合蛋白的表达定位。结果:荧光显微镜观察显示,BPI-EGFP融合蛋白分布在整个细胞质中,并且在核膜周围有高表达。PCR扩增证实了BPIcDNA片段和EGFP片段已整合入MCF-7细胞基因组中。RT-PCR证明了hBPI和EGFP的融合蛋白在MCF-7细胞中在转录水平的表达。Western blot分析显示hBPI-EGFP以定位于细胞质和分泌到MCF-7细胞外两种形式表达。结论:hBPI-EGFP融合蛋白与天然的嗜中性的多核粒细胞中的定位和转运特点是一致的。

综述了最近根癌农杆菌介导不同宿主遗传转化的转化机理和T-DNA的整合方式等方面的研究进展及目前对农杆菌介导的遗传转化中植物、真菌以及人类细胞宿主的研究进展.

The leaves of Saussurea involucrata as the explants were cultured and induced to form calli on the media prepared according to different formulae;The calli were cultured and induced to form somatic embryos,which were cultured to differentiate into regeneration plantlets of S.involucrata.It was shown that the optimal medium for callus induction was MS+0.5 mg/L 2,4-D+1.5 mg/L BA and its callus induction rate was as high as 100%;The calli were transferred and secondarily cultured on the medium of MS+0.5 mg/L 2,4-D+1.5 mg/L BA and then the propagated calluses were transferred,cultured and induced to form somatic embryos of S.involucrata on the liquid medium of MS+0.2 mg/L 2,4-D and the somatic embryo induction rate was as high as 40%;The somatic embryos were inoculated on the medium of MS+0.5 mg/L ABA and then differentiated into great numbers of regeneration plantlets of S.involucrata.

多孔菌科菌类层孔菌属(Fomes)真菌,全世界约23余种,在我国均有分布,是一种常见的真菌类.一些种在国内外民间被广泛作为药用,如药用层孔菌(F. officinalis)用于治疗咳嗽、痰喘、胃痛、石淋、咽喉痛、牙周炎及毒蛇咬伤,亦用于镇惊、祛风、止血、止痛等功效[1];多年层孔菌(F.annosa)、红缘层孔菌(F. cytisina)等均含有抗癌物质,对小鼠肉瘤S-180有一定抑制作用.

By the orthogonalty design, the factors influencing suspension cells growth of L.angustifolia were studied, and best culture conditions which were B5 media supplemented with 40 g/L sucrose, 0.025 mg/L 2,4- D, 0.5 mg/L 6-BA and the rotating speed of 120 r/min were sifted. After 15 days of culture, the density of surviving cells could reach 4.4×105 cell/ml, and differentiation rate of suspension cells could surpass 90% after 3 months of suspension culture. The cause of effects of sucrose concentration on the density of surviving cells was discussed through the analysis of the consumption rule of surplus sucrose.

通过研究接种量、培养基、碳源、氮源及激素组合等生物学因子对薰衣草胚性愈伤组织悬浮培养生长的影响,建立了一种可快速生长的薰衣草胚性愈伤组织悬浮培养体系.结果表明,在添加0.05 mg/L 2,4-D、0.5 mg/L 6-BA、30 g/L蔗糖及0.5 g/L水解酪蛋白的MS培养液中,以50 g/L的接种量进行薰衣草胚性愈伤组织的悬浮培养,其生长速率可达24.14 g/(L·d).

介绍植物抗寒冻基因研究中一些已分离和鉴定出的低温诱导表达基因及其抗寒功能、低温信号转导以及调控方式的研究进展.

概述了植物体细胞无性系变异的类型、机制及在植物性状改良中的应用,并着重对无性系变异的筛选方法和用组织培养中出现的变异进行植物性状改良作了简要介绍.