Shoot organogenesis from cultured seed explants of peanut (Arachis hypogaea L.) using thidiazuron

Summary Thidiazuron (TDZ) was utilized to induce adventitious shoot formation from the hypocotyl region of cultured seed explants of peanut (Arachis hypogaea L.). Excision of the radicle from seed explants was more stimulatory to shoot initiation than removal of the epicotyl alone. Removal of both the radicle and the epicotyl from seeds resulted in a 37-fold increase in the frequency of shoot production when compared to intact seeds. Half seed explants with epicotyl and radicle removed produced the greatest number of shoots per explant. Explants from mature seeds were more responsive to TDZ than immature seed-derived explants. A 1-wk exposure to 10 μM TDZ was sufficient to stimulate the initiation of adventitious shoots that subsequently developed into plants. High frequency of shoot initiation was readily induced in a variety of genotypes ofA. hypogaea and a wild peanut (A. glabrata). Plants regenerated from shoots induced by TDZ were phenotypically normal and fertile.     

An improved procedure for protoplast microelectrofusion and culture of Nicotiana tabacum intraspecific somatic hybrids: plant regeneration and initial proofs on organelle segregation

Somatic hybrids were produced between haploid Nicotiana tabacum L. cv. Petite Havana (wild type) and haploid streptomycin resistant (SR1) mutant by an improved version of microelectrofusion of preselected pairs of protoplasts and the culture of fusion products in a nurse culture. Resistance of diploid plants regenerated from 20 somatic hybrid clones was tested at low concentration of streptomycin in the light as well as at high concentrations of streptomycin in the dark. In two independent hybrid lines, plants resistant in the light but sensitive in the dark were found. The existence of this plant type indicates a segregation of chloroplasts and mitocondria in somatic hybrid clones. It is suggested that microelectrofusion of preselected pairs of protoplasts combined with a reliable nurse culture might be a good technique for controlled somatic hybridization, cell reconstitution and partial gene transfer to different plant species. It might also be used to follow and analyse organelle segregation in somatic hybrid clones. The possibility that mitochondria might be resistant to streptomycin in the SR1 mutant is also discussed.     

无血清原代培养大鼠海马神经元的形态与膜电位

分离新生Wistar鼠海马,采用添加B27的无血清培养液进行海马神经元原代培养,动态观察海马神经元形态学变化;通过免疫荧光细胞化学法检测神经纤丝(NF)的表达,进行神经元鉴定及纯度计算;采用电位敏感的荧光探针标记神经元,在激光扫描共聚焦显微镜上动态监测去极化剂KCl作用前后膜电位的变化,观察神经元电生理反应。结果表明:此方法培养的大鼠海马神经元可在体外存活20天以上,9～14天为发育最成熟阶段,培养7天神经元纯度达90%。KCl作用于细胞后胞内荧光强度增强,细胞迅速去极化。本培养方法在体外获得高纯度的海马神经元并延长体外存活时间,且显示出神经元的电生理反应特性。     

东北森林净第一性生产力与碳收支对气候变化的响应

以东北地区(38.43'N～53.34'N,115.37'E～135.5'E)为研究对象,利用当前气候状况和不同气候情景下的气象数据驱动基于个体生长过程的中国森林生态系统碳收支模型FORCCHN,模拟了气候变化对东北森林生态系统净第一性生产力(NPP)和碳收支(NEP)的影响.结果表明:1981～2002年期间,东北森林NPP总量位于0.27～0.40 pgc·a-1之间,平均值为0.34 pgc·a-1;土壤呼吸总量在0.11～0.27 PgC·a-1,平均为0.19 PgC·a-1;NEP总量位于0.11～0.18 PgC·a-1之间,且近20多年来该区森林起着CO2汇的作用,平均每年吸收0.15 Pg C的CO2;该区森林NPP和NEP对温度升高比对降雨变化的反应更为敏感;综合降雨增加(20%)和气温增加(3℃)的情况,该区各点森林的NPP和NEP增加的幅度最大;温度不变、降水增加(不变)情景下最小.     

华南地区龙眼的温度适宜性及其变化趋势

运用温度适宜度模型对华南地区龙眼生产和生育期的适宜性及其时空差异进行了计算、评价和预测。首先分析龙眼各生育期适宜度,结果表明,休眠期、花芽生理分化期、花芽形态分化期适宜性较低,具有较高的变率,开花期、果实发育成熟期、抽梢期表现出较高的温度适宜性,变率也较小,并且通过进一步研究发现影响华南地区龙眼生产的关键问题不是冷害,而是冬春季节的热害;其次分析华南地区各县区的平均温度适宜度,结果表明,华南地区龙眼温度适宜性的地域分异呈纬度地带性规律,并根据龙眼减产率将华南地区划分为最适宜型、适宜型、次适宜型、低适宜型和不适宜型五大适宜度类型;最后分析龙眼各生育期温度适宜度的年际变化,发现未来各生育期中除果实发育成熟期和抽梢期无明显变动外,其他生育期的温度适宜度都有下降趋势,同时对华南地区各站点龙眼温度适宜度的变化趋势作分析,结果表明,除福建省部分地区有弱上升外,总体上华南地区温度适宜度有下降趋势,且存在着地区差异,并且未来龙眼的适宜种植区将有可能北移,根据各站点未来适宜度变化的方向和强度进行分类,将华南地区划分为强下降型、次强下降型、弱下降型和上升型四大变动类型。     

高效抗氧化大型真菌的筛选

为获得高效抗氧化菌株,采用直接提取方式从20个大型真菌菌株菌丝体培养液中提取抗氧化活性物质,采用邻苯三酚自氧化法、水杨酸法、DPPH法测定各菌株菌丝体培养粗提液对超氧阴离子自由基(O2)、羟基自由基.(OH)及1,1-二苯代苦味酰基自由基(DPPH-)的清除作用。结果表明:各菌株菌丝体培养粗提液对(O2)、.OH及DPPH-均有一定的清除作用,其中菌株NG菌丝体培养粗提液对OH-的清除效果最好,清除率为75.56%;菌株02菌丝体培养粗提液对(O2-)的清除效果最好,清除率为37.51%;菌株EG菌丝体培养粗提液对DPPH-清除效果最好,清除率为66.91%。     

阳桃胚乳愈伤组织诱导和不定芽发生的研究

首次成功建立阳桃胚乳组织培养并获得胚乳再生植株。胚乳愈伤组织诱导以培养基MS 2,4-D2．0mgL^-1 BA0．2mgL^-1的效果最好,诱导频率可达94．7％,愈伤组织乳白色,结构致密,生长旺盛;将其接种在培养基MS ZT3．0mgL^-1 NAA0．2mgL^-1上,愈伤组织由乳白色致密型转变为淡绿色致密型,进而形成绿色芽点,分化出不定芽,分化频率可达73．3％;胚乳植株在培养基MS ZT2．0-2．5mgL^-1 NAA0．05mgL^-1上进行壮苗和营养繁殖。     

云杉属树种的体细胞胚胎发生

综述了云杉属树种体细胞胚胎发生的研究现状,其中包括：（１）影响云杉属树种体细胞胚胎发生及其植株再生的因素;（２）云杉属树种体细胞胚胎发生的形态学和细胞组织学研究。并展望了云杉属树种体细胞胚胎发生的应用前景及研究方向。     

Transient Transfection Factors for High-Level Recombinant Protein Production in Suspension Cultured Mammalian Cells

The efficient transfection of cloned genes into mammalian cells system plays a critical role in the production of large quantities of recombinant proteins (r-proteins). In order to establish a simple and scaleable transient protein production system, we have used a cationic lipid-based transfection reagent-FreeStyle MAX to study transient transfection in serum-free suspension human embryonic kidney (HEK) 293 and Chinese hamster ovary (CHO) cells. We used quantification of green fluorescent protein (GFP) to monitor transfection efficiency and expression of a cloned human IgG antibody to monitor r-protein production. Parameters including transfection reagent concentration, DNA concentration, the time of complex formation, and the cell density at the time of transfection were analyzed and optimized. About 70% GFP-positive cells and 50-80 mg/l of secreted IgG antibody were obtained in both HEK-293 and CHO cells under optimal conditions. Scale-up of the transfection system to 1 l resulted in similar transfection efficiency and protein production. In addition, we evaluated production of therapeutic proteins such as human erythropoietin and human blood coagulation factor IX in both HEK-293 and CHO cells. Our results showed that the higher quantity of protein production was obtained by using optimal transient transfection conditions in serum-free adapted suspension mammalian cells.     

Investigating the long‐term legacy of drought and warming on the soil microbial community across five European shrubland ecosystems

We investigated how the legacy of warming and summer drought affected microbial communities in five different replicated long‐term (>10 years) field experiments across Europe (EU‐FP7 INCREASE infrastructure). To focus explicitly on legacy effects (i.e., indirect rather than direct effects of the environmental factors), we measured microbial variables under the same moisture and temperature in a brief screening, and following a pre‐incubation at stable conditions. Specifically, we investigated the size and composition of the soil microbial community (PLFA) alongside measurements of bacterial (leucine incorporation) and fungal (acetate in ergosterol incorporation) growth rates, previously shown to be highly responsive to changes in environmental factors, and microbial respiration. We found no legacy effects on the microbial community size, composition, growth rates, or basal respiration rates at the effect sizes used in our experimental setup (0.6 °C, about 30% precipitation reduction). Our findings support previous reports from single short‐term ecosystem studies thereby providing a clear evidence base to allow long‐term, broad‐scale generalizations to be made. The implication of our study is that warming and summer drought will not result in legacy effects on the microbial community and their processes within the effect sizes here studied. While legacy effects on microbial processes during perturbation cycles, such as drying–rewetting, and on tolerance to drought and warming remain to be studied, our results suggest that any effects on overall ecosystem processes will be rather limited. Thus, the legacies of warming and drought should not be prioritized factors to consider when modeling contemporary rates of biogeochemical processes in soil.