干柱层析法制备油茶总皂苷对照品初探

目的 研究干柱层析法制备油茶皂苷对照品的方法。方法 原料用60 %乙醇超声提取,浓缩至无醇味,石油醚脱酯后用正丁醇按3:1体积比加入萃取。取正丁醇层并真空浓缩至干,干法上样于100目~200目硅胶柱色谱分离,用乙酸乙酯-甲醇-正丁醇-水(2:1:4:7)的上相展层;根据Rf值取出相应的条带并用甲醇淋洗收集蒸干,薄层及理化性质检测。结果 该方法所制样品由薄层色谱为一点,纯度较高。结论 该方法简便、实用且制备量大,可作为油茶总皂苷对照品。     

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.     

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。     

Polygalacturonase and β-galactosidase activities in Hayward kiwifruit as affected by light exposure,maturity stage and storage time

Softening process and total antioxidant activity were evaluated in kiwifruit differently exposed to light intensity and harvested at two different times (October 10 and November 14). Fruit was stored for 2 months at 0 °C (S1) and then maintained for a week at ambient temperature (S2). The results showed that fruit harvested later presented a faster softening rate during storage than fruit harvested earlier, even if antioxidant activity did not change. Light-exposed fruit showed higher flesh firmness than that of shaded ones. Polygalacturonase activity was higher in kiwifruit maintained for a week at ambient temperature after cool storage and, in particular the highest value was recorded in fruit harvested later. The behaviour of β-galactosidase was different: it did not show changes in fruit harvested later and significantly decreased in light- and shade-exposed fruit harvested earlier.     

Evaluating foliar nitrogen compounds as indicators of nitrogen status in Prunus persica trees

Mobile nitrogen (N) forms may be better N indicators of the N status of trees than total nitrogen (TN) due to their higher sensitivity to increasing N supply. A field experiment was carried out over a 3-year period to compare foliar concentrations of total N (TN), soluble N (SN), chlorophyll (Minolta SPAD readings), NH₄–N and NO₃–N as indicators of soil N availability in nectarine, Prunus persica L. Batsch, cv. ‘Fantasia’ (grafted on ‘Nemaguard’ peach, P. persica × P. davidiana) trees. Young trees were exposed to a range of fertilizer-N application rates. Based on correlation analysis, the best association between leaf N compounds with soil N supply and trunk diameter and/or fruit yield was obtained with TN and chlorophyll SPAD readings. Leaf concentrations of mobile N compounds (NH₄–N and NO₃–N) increased more than any other N compound under high N supply; however, their inconsistency among years and low leaf concentration difficult their use as N indicators. The optimum foliar TN for growth decreased with tree age, 4.4%, 3.6% and 3.3% in non-bearing 1-year-old trees, non-bearing 2-year-old trees and 3.3 fruit-bearing 3-year-old trees. The optimum SPAD readings were 40 in 2-year-old trees and 42 in 3-year-old trees. Stable N compounds (TN and chlorophyll SPAD) could be used to N diagnosis in the zone of N deficiency, and soluble N compounds (NH₄–N and NO₃–N) to diagnoses N excess.     

利用DAD1反义片段转化创建菜薹可调控雄性不育材料

 菜薹因为没有好的雄性不育材料或自交不亲和系,至今尚无一代杂种用于生产,根据拟南芥及白菜型油菜的花药不开裂基因DAD1的保守序列设计引物,扩增菜薹的DAD1基因片段（DAD1F）,构建反义DAD1F植物表达载体,用农杆菌介导法转化菜薹,对转基因植株进行分子检测,鉴定其雄性不育性并进行育性恢复试验.克隆得到的菜薹的DAD1基因片段大小为678 bp,命名为BrcpDAD1F,其序列与拟南芥和白菜型油菜的DAD1高度同源,同源率分别为88％和99％;共得到了12株转基因植株,有6株在转录水平上得到表达,表现为雄性不育,花器官畸形,花粉活力低,萌发率不到10％,且开花后不能结角果或结空角果,或者得到极少种子但种子不萌发;用对照的花粉给转基因植株授粉可使其正常结实.以500 μmol·L^-1茉莉酸甲酯处理可使其雄性不育得到恢复,花粉可以在柱头和培养基上萌发,具有受精能力。T₁代可育株与不育株的比例都呈1:3分离, T₂代不同株系的育性分离比例不同,有些株系继续呈1:3的分离,有些株系全是可育株或全是不育株,说明反义抑制呈单基因稳定遗传。     

不同类型甜菜品种根中Mg^2＋-ATP酶活性和产质量的关系

测定不同类型甜菜品种各生育时期根中Mg^2＋-ATP酶活性、可溶性总糖、可溶性蛋白含量和产量、含糖率。结果表明,在幼苗期和叶丛繁茂期,Mg^2＋-ATP活性呈先上升后下降趋势,从块根增长期直至糖分积累期,Mg^2＋-ATP酶的活性逐渐增强,并保持在全生育期最高水平。不同类型品种间存在差异,在各生育时期高糖型品种Mg^2＋-ATP酶活性较强。根中Mg^2＋-ATP酶活性与根中可溶性总糖呈正相关,与甜菜根的成熟进程正相关;Mg^2＋-ATP酶活性与可溶性蛋白质舍量呈负相关,对可溶性糖的积累方面,二者的作用刚好相反。高糖型品种的可溶性总糖含量较高．相应的根中蛋白质含量很低。Mg^2＋-ATP酶活性较高．