Genomic analyses reveal natural selection on reproduction related genes between two closely related Populus (Salicaceae) species

Identifying the factors that cause reproductive isolation and their relative importance in species divergence is crucial to our understanding of speciation processes. In most species, natural selection is commonly considered to play a large role in driving speciation. Based on whole genome re-sequencing data from 27 Populus alba and 28 Populus adenopoda individuals, we explored the factors related to reproductive isolation of these two closely related species. The results showed that the two species diverged ~5–10 million years ago (Ma), when the Qinghai–Tibet Plateau reached a certain height and the inland climate of the Asian continent became arid. In highly differentiated genomic regions, the relative divergence (F_ST) and absolute divergence (d_xy) were significantly higher than the genomic background, θπ and shared polymorphisms decreased whereas fixed differences increased, which indicated that natural selection played a key role in the reproductive isolation of the two species. In addition, we found several genes that were related to reproduction that may be involved in explaining the reproductive isolation. Using phylogenetic trees resolved from haplotype data of Populus tomentosa and P. adenopoda, the maternal origin of P. tomentosa from P. adenopoda was likely to be located in Hubei and Chongqing Provinces.     

Poplar (Populus nigra L.) plants transformed with aBacillus thuringiensis toxin gene: insecticidal activity and genomic analysis

Insect-resistant poplar (Populus nigra L.) plants have been produced by infecting leaves withAgrobacterium tumefaciens strains carrying a binary vector containing different truncated forms of aBacillus thuringiensis (B.t.) toxin gene under a duplicated CaMV 35S promoter. Putative transgenic plants were propagated by cuttings at two experimental farms (in Beijing and Xinjiang, China). At 2–3 years after transformation, 17 of them were selected on the bases of insect-tolerance and good silvicultural traits, and evaluated for insect resistance, for the presence of theB.t. toxin DNA fragment (Southern blots and PCR) and for the expression of the transgene (western and northern blots). Somaclonal variation, as suggested by the appearance of permanent changes in the shape of the leaves, was also investigated with molecular tools (RFLP (restriction fragment length polymorphism), RAPD (random amplified polymorphic DNA) and microsatellite DNA).Bioassays withApochemia cineraius andLymantria dispar on the leaves of the selected clones showed different and, in some cases, high levels of insecticidal activity. The molecular analysis demonstrated integration and expression of the foreign gene. Somatic changes were correlated to extensive genomic changes and were quantified in dendrograms, in terms of genomic similarity. The analysis of control plants suggested that genomic changes were correlated to thein vitro culture step necessary forA. tumefaciens-mediated gene transfer, rather than to the integration of the foreign genes.Three transgenic clones (12, 153 and 192), selected for insect resistance, reduced morphological changes and promising silvicultural traits, are now under large-scale field evaluation in six different provinces in China.     

黄芪总黄酮对DNA损伤防护作用的研究

用DNA解旋荧光检测法(FADU)研究了黄芪总黄酮(TFA)对γ射线和H₂O₂所致V79细胞DNA链断裂的防护作用. 结果表明TFA对这两种损伤因子所致的DNA损伤均有不同程度的防护作用, 当TFA浓度达到0.4g/L和0.6g/L时, 分别对H₂O₂和γ射线所致的损伤有保护作用(P<0.05), 而浓度增至0.8g/L和1.2g/L时, 分别对两种因素所致的DNA链断裂损伤有非常显著的防护效果(P<0.01), 对H₂O₂的防护效果优于对γ射线.     

超氧化物歧化酶模拟化合物的生物活性研究

根据天然超氧化物歧化酶(SOD)活性部位结构合成了含苯并咪唑的5种配体及其32种分别含Cu（Ⅱ）、Fe（Ⅲ）、Mn（Ⅱ）、Co（Ⅱ）的模拟化合物.经光谱、电化学测试证明这些化合物具有拟S3D活性,其50％抑制率浓度（IC50）为10-6～10-8mol·L-1,催化超氧离子自由基（O-·2）歧化的反应速率常数kq为106～108mol-1·L·s-1.同时观察到几种模拟化合物有抗肿瘤活性或增强水稻抗寒性.     

全自动生化分析仪测定血清AST同工酶

应用天冬氨酸氨基转移酶抑制剂“AMANO-3”水解样品中的线粒体型天冬氨酸氨基转移酶同工酶（m-AST）,测定血清中剩余的胞浆型AST同工酶（c-AST）活性,进而与总AST活性相比较并计算出受抑制剂水解的m-AST同工酶活性．由于此方法采用蛋白水解反应破坏m-AST,因此测定方法可直接应用于生化自动分析仪．亦建立了一个适于日立7150分析仪的AST同工酶联合测定方法．其测定和计算出的m-AST同工酶批内CV为3.5％～7.9％;其结果(y)与AST同工酶电泳迁移率(x)相关．y=1.019x-0.489,r＝0.996（n＝30）．测定了113名健康人m-AST和c-AST,其m-AST参考值范围1.64～9.64U/L,x±s＝(5.641±2.013)U/L;c-AST参考值范围5.69～16.81U/L,x±s＝(5.641±2.013)U/L．     

脏器微血管对荧光素钠通透性的实验方法

大鼠颈动脉注射１％ＦｌＮａ,荧光显微镜下活体观察肠系膜微血管血流状态及ＦｌＮａ的渗出情况,并在不同时间点经股动脉采血,测定血浆内ＦｌＮａ浓度随时间的变化,利用组织匀浆测定不同脏器中ＦｌＮａ的分布,再辅以冰冻切片进行观察。活体观察发现,ＦｌＮａ注入体内后,经微血管迅速向周围组织渗出,最后汇集于淋巴管,血浆及组织匀浆ＦｌＮａ浓度的测定表明,ＦｌＮａ浓度随时间的变化呈指数衰减,各脏器ＦｌＮａ的分布极不相同。冰冻切片也显示了同样的分布差别。这些结果表明,我们所建立的方法可直观、定量地反映ＦｌＮａ在微血管的通透情况。     

黄芪有效成分对氧自由基清除作用的ESR研究

用电子自旋共振技术研究了黄芪总黄酮(TFA)、黄芪总皂甙(TSA)和黄芪总多糖(TPA)对次黄嘌呤／黄嘌呤氧化酶体系产生的超氧阴离子自由基和H₂O₂－Fe²⁺体系产生的羟自由基的清除作用．结果表明,这3种成分均有清除氧自由基的作用;对超氧阴离子自由基的清除效能大于对羟自由基的清除作用;其作用强度依次为TFA＞TSA＞TPA．结果提示清除氧自由基可能是黄芪抗衰老的主要机理之一,TFA和TSA是黄芪抗氧化作用的主要药理活性成分．     

适用于获取最优化配方的一种算法

本文应用印楝种仁提取物（F3）与敌敌畏混配为例,以斜纹夜蛾（Spodopteralitura）为目标害虫,介绍一种适用于获取最优化配方的算法,在二次通用回归旋转组合设计的基础上,经参数辨识,获取二次回归方程,经失拟性、回归显著性检验,本方程基本能够反映杀虫剂用量与斜纹夜蛾幼虫死亡机率值之间的关系．在害虫防治实践中,要求在防治费用最小的基础上,目标害虫有最大的死亡率.因此,以防治目标害虫的费用作为优化算法的目标函数,以害虫死亡机率值最大作为约束条件,有如下的一组优化算式为目标函数约束条件式中a1,a2分别为参试杀虫剂1,2最低用量,b1,b2则为相应的最高用量．C1,C2分别为杀虫剂1,2的单价,N1,N2为杀虫剂l,2的用量．Y为目标害虫死亡机率值回归方程．本文所依据的试验设计中,以对数函数关系变换编码值与使用浓度之间的关系,所以应用拉格朗日求极值原理求取最优化配方．由计算所得的混配比例与其他方法所获结果一致．     

角结膜缘上皮良、恶性病变细胞核DNA原位图像定量分析

应用真彩色医学图像分析技术,对５１例角结膜缘上皮良、恶性病变（其中上皮增生１９例,不典型增生６例,原位癌１４例,鳞状细胞癌１２例）进行了ＤＮＡ原位定量分析研究．结果显示：原位癌和鳞状细胞癌ＤＮＡ含量明显增加,多为高倍异倍体细胞,ＤＮＡ直方圆明显右移,峰值主要位于＞５Ｃ处,≥５Ｃ细胞分别占７８．８９％和７８．３１％;上度增生病变ＤＮＡ含量较低,多为低倍整倍体细胞,ＤＮＡ直方图峰值位于２Ｃ～４Ｃ处,２Ｃ～４Ｃ细胞占８８．５９％;上皮不典型增生病变ＤＮＡ含量介于良性病变和癌之间,ＤＮＡ直方图逐渐右移,２Ｃ～４Ｃ细胞占５８．６２％,≥５Ｃ细胞占４１．３８％。以上数据经统计学处理各组间有显著性差异。表明ＤＮＡ倍性程度与肿瘤的增殖程度呈正相关,高倍异倍体细胞随肿瘤恶性程度的增高而增多。作者认为ＤＮＡ原位图像定量分析可为角结膜缘上皮良、恶性病变的诊断、分级及早期发现癌变趋势提供一个可靠的参考指标。     

中药金樱子的研究应用概况

本文就国内外对中药金樱子的化学成分及其提取分离方法、药理学研究和临麻应用作了综述,为金樱子的综合开发提供依据。