百合总RNA提取方法的比较和分析

以卷丹、麝香百合品种富田和离体鲜切花的花蕾为材料,比较Trizol法、异硫氰酸胍法、CTAB改进法和SDS改进法提取总RNA的效果,结果表明,SDS改进法能有效去除多糖,提取的RNA中28S rRNA亮度约为18S rRNA的两倍,OD260/OD280值介于1．7-2．2之间,卷丹RNA得率为132．52μg／g,富田RNA得率为186．88μg／g。进一步用SDS改进法分别提取富田外轮花瓣、内轮花瓣、雄蕊、雌蕊、叶和茎的RNA,同样可以获得完整的纯度高的RNA,其中28S rRNA亮度约为18S rRNA的两倍,OD260／0D280值介于1．7～2．2之间,说明此方法适用于百合各个组织RNA的提取。经RT—PCR获得了花发育基因的特异性条带,说明用SDS改进法从百合中提取的RNA质量好、产率高、完整性强,完全适合于百合进一步的分子生物学研究。     

烟草不同组织总RNA的提取方法初探

【研究目的】寻找烟草各个组织总RNA最佳的提取方法。【方法】以烟草K326的根、茎、叶、蕾、花和种子为材料,分别采用试剂盒法、Trizol法和CTAB法提取烟草6个组织总RNA进行对比。【结果】烟草不同组织总RNA提取方法有所不同,试剂盒法和Trizol法只能提取出根、茎、叶、蕾和花的总RNA,而不能提取种子总RNA;CTAB法能提取出6个组织的RNA。3种方法提取的RNA质量好,均能满足RT-PCR以及后续实验的要求。【结论】试剂盒法较Trizol法和CTAB法简单、快捷,适合于烟草除种子以外其余组织总RNA快速提取,CTAB法适合烟草所有组织总RNA提取。     

棉铃虫HaHR3基因的克隆及其植物RNA干扰载体的构建

采用RT-PCR方法克隆了棉铃虫蜕皮调节转录因子HaHR3基因,该基因含有1671 bp的完整开放阅读框。序列分析表明,HaHR3与多种昆虫蜕皮调节转录因子高度同源。利用生物信息学方法对HaHR3的结构特征进行分析发现,HaHR3具有蜕皮调节转录因子超家族的典型特征,包括两个锌指结构和一个DNA结合结构域,不存在信号肽序列和N糖基化位点。选择HaHR3基因的部分片段构建了RNAi中间载体pRNAi1017-HaHR3sa,再将HaHR3正反向序列亚克隆至植物表达载体pCAMBIA2300-35S-OCS,成功构建了由35s启动子调控的HaHR3基因的正反向RNA干扰载体pCAM-RNAi-HaHR3。这一载体的成功构建为下一步通过植物介导的RNA干扰技术防治棉铃虫打下了坚实的基础。     

棉花RNA的快速提取方法

目前已发展的从植物中提取RNA的方法有多种.但由于棉花中次生物质含量较高,因此有些方法并不适合于棉花RNA的提取.本文介绍经过反复实验后改进的棉花RNA的提取方法,它具有快速、简单等特点.     

Fractal features of soil particle size distribution and the implication for indicating desertification

Some of the effects of land desertification on soil properties are manifested by the coarsening of the soil particlesize distribution (PSD) and the losses in organic C and nutrients. The changes and characteristics of PSD and selected chemical properties in soils at the 0-15 cm plough layer from different degrees of desertified croplands are analyzed in the semiarid Horqin Sandy Land, northern China. The fractal dimension of the PSD is emanated to characterize the patterns of PSD. The relationships between the fractal dimension of the PSD and selected soil properties are discussed. The results show that: (1) in the transformation from potential desertified cropland to extremely desertified cropland, the sand content at the 0-15 cm soil increased from 69% to 93%, organic C and total N contents decreased by 65% and 69%, respectively; (2) the fractal dimension of PSD ranged from 2.179 to 2.611, the more the contents of sand, the lower the fractal dimension and the higher the desertified degree of farmland. In the desertification process within the studied area, the mean fractal dimension decreased from 2.555 for the potential desertified soils to 2.298 for extremely desertified soils; (3) there existed considerable linear relationships between fractal dimension and soil properties. It was shown that fractal dimensions of PSD are useful parameters able to monitor soil degradation and to estimate the degree of soil desertification.     

土壤沉积物样品中有机碳含量的快速测定

采用微波密闭消解样品的方法来测定土壤／沉积物中有机碳的含量。与重铬酸钾氧化法的结果相比，微波消化法具有操作简单、污染小、快速准确的优点。经F检验法及t检验法检验，两种方法的分析结果无明显差异。     

用点杂交方法定量检测植物病毒RNA负荷量

用RNA点杂交与用放射性成像仪直接读取放射性活度相结合的方法定量测定植物病毒RNA相对负荷量 ,检测了 0 0 0 1～ 1 0ppm浓度范围内二氢茉莉酸丙酯(PDJ)对烟草组织中烟草花叶病毒 (TMV)RNA的影响。结果表明 :在PDJ处理 3d时 ,与对照相比低浓度 ( 0 0 0 1ppm)PDJ处理的TMVRNA病毒负荷量无明显差异 ,但随着浓度的升高 ,PDJ对TMVRNA的复制和积累具有促进作用 ;浓度越高 ,促进作用越显著。同时用传统的枯斑接种法验证了上述结果。两种方法相比 ,点杂交结合放射性成像仪读取放射性活度的方法所受的人为误差干扰较小 ,能够相对精确地定量测定植物组织中病毒RNA负荷量。     

Tillage and crop sequence effects on organic carbon and total nitrogen content in an irrigated Alberta soil

A better understanding of tillage effects on soil organic matter is vital for development of effective soil conservation practices. The objective of this research is to determine the effect of tillage and crop sequence on soil organic carbon (OC) and total nitrogen (TN) content in an irrigated southern Alberta soil. A field experiment was conducted using a split–split plot design from 1994 to 1998 in Alberta, Canada. There were two crop sequences (Sequence 1: spring wheat (Triticum aestivum L.)–sugar beet (Beta vulgaris L.)–spring wheat–annual legume; and Sequence 2: spring wheat–spring wheat–annual legume–sugar beet) and two tillage practices (CT: conventional tillage and MT: minimum tillage). Surface soil under MT had significantly higher OC (30.1 Mg ha⁻¹) content than under CT (28.3 Mg ha⁻¹) after 4 years of treatment. The MT treatment retains crop residue at the soil surface, reduces soil erosion and slows organic matter decomposition, which are key factors in enhancing the soil fertility status of southern Alberta irrigated soils.     

青钱柳叶总黄酮超声辅助提取工艺优

为研究以乙醇为溶剂超声波辅助提取青钱柳叶总黄酮的工艺,在单因素试验的基础上,采用四因素二次回归正交旋转组合设计,以液料比、超声处理时间、超声功率、溶剂体积分数为考察因素,优选了超声波辅助提取青钱柳叶总黄酮的最佳工艺.结果表明各因素对提取率的影响大小依次为:超声处理时间、溶剂体积分数、液料比、超声功率.最佳工艺条件为液料比55mL/g,超声处理时间34min,超声功率720W,溶剂体积分数60%,在此最佳条件下,总黄酮的得率为86.63%,优化工艺稳定,提取率也较高,试验验证结果与模型预测值相符.     

低磷胁迫对玉米叶片色素和形态的影响

通过大田试验,以4个基因型玉米为材料,研究了低磷胁迫下玉米叶片色素和形态变化及其内部生理机制.结果表明,低磷条件下,玉米叶片叶绿素/类胡萝卜素、叶绿素/紫色素和叶绿素/红色素均减少,其中基因型"7922"和"掖107"的叶绿素/类胡萝卜素、叶绿素/紫色素和叶绿素,红色素减少幅度较大,为高磷条件下的0.4～0.8倍,且减少幅度高于"178"和"*082".低磷条件下,苯丙氨酸解氨酶(PAL)活性和总黄酮含量均增加,其中基因型"7922"和"掖107"为高磷条件下的1.5～2.0倍,增加幅度高于基因型"178"和"*082"."7922"和"掖107"的叶宽和鲜重在两个供磷水平下差异显著,"掖107"的叶片主叶脉直径和总叶片数差异显著.低磷土壤条件下玉米叶宽和主叶脉直径变小,鲜重和总叶片数减少.不同基因型玉米叶片色素和形态变化差异显著.初步推测,"7922"和"掖107"不耐低磷, "178"和"*082"较耐低磷.