利用shRNA真核表达载体干涉雌性鸡胚性腺中芳香化酶基因(CYP19A1)的表达

本研究通过把构建的短发夹结构RNA(shRNA)真核表达载体导入鸡胚,检测鸡胚性腺分化期质粒载体在鸡胚体内代谢情况及雌性鸡胚性腺中芳香化酶基因(CYP19A1)mRNA表达效率,进而探讨利用该方法在鸡胚体内进行特定基因干涉的可行性.实验针对CYP19A1基因构建了4条特异性表达载体,一条非特异性表达载体.每个实验组选取45个新鲜种蛋作为实验材料进行胚盘下腔注射,并设立空白对照组.鸡胚发育12d时,检测各处理组鸡胚肝脏组织中质粒存在情况,并取其左侧性腺组织进行目标基因的荧光定量分析.研究结果表明,导入组在12日胚龄时所有鸡胚基因组中均可检测到绿色荧光蛋白基因(EGFP);荧光定量结果显示,导入特异性表达载体cyp-580、cyp-1083和cyp-1295后,对应雌性鸡胚性腺CYP19A1 mRNA表达效率显著低于空白对照组,干涉效率分别为:73％、52％和85％;特异性表达载体cyp-1403组CYP19A1mRNA表达效率与空白对照组相比有所降低但无显著性差异.本实验为诱导鸡胚性反转提供了新方法并建立了鸡胚发育期特定基因体内干涉新平台.     

不同改良剂对铬污染土壤中小白菜生理、生化特性的影响

为了研究不同种类改良剂对铬污染土壤中小白菜生理生化特性的影响,采用盆栽试验监测在不同浓度的铬污染的土壤中小白菜(Brassica parichinensis)的叶绿素含量、超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性以及丙二醛(MDA)含量的变化。研究发现,不同浓度的铬污染土壤中,3种改良剂添加使叶绿素含量及SOD酶活性发生变化并高于对照组而POD酶活性降低,使铬对CAT酶活性的抑制作用得到缓解。石灰、石灰+活性炭用量的增加使MDA含量先降低后升高,而活性炭的施入减少MDA的含量。通过比较,3种改良剂的改良效果为活性炭>石灰+活性炭>石灰。     

基于关键气象因子的中稻单产动态预报

利用重庆市1986-2009年中稻单产和中稻生育期内旬平均气温、旬降水量、旬日照时数等资料,采用统计分析法建立中稻气象产量动态预报模型,在此基础上预报中稻单产。用5a滑动平均方法分离中稻趋势产量,在分析气象产量与中稻全生育期逐旬气象要素相关性的基础上,以3月中旬平均气温、5月下旬日照时数、7月中旬平均气温、7月中旬降水量、7月中旬日照时数、7月下旬平均气温、7月下旬降水量和8月上旬平均气温等作为关键气象因子,建立中稻气象产量动态预报模型,并应用该模型实现中稻单产动态预报。对1986-2009年的中稻单产做模拟检验,平均准确率在96%以上,95%以上的年份预报准确率超过90%。对2010年的单产进行预报,准确率为91.5%～92.8%,预报准确率较高,基本能满足业务服务的需要。     

长期施肥和秸秆还田对红壤水稻土氮素分布和矿化特性的影响

为明确长期施肥和秸秆还田对稻田土壤生态系统氮素循环过程的影响,探索维持稻田土壤长期供氮潜力的途径,利用始于1990年的湖南红壤水稻土长期定位试验,研究了长期施肥和秸秆还田对水稻土剖面氮素迁移分布和C/N的影响,比较了不同温度下土壤氮素矿化曲线变化和不同施肥方式氮素矿化动态差异,分析了氮素累积矿化量与有效积温的关系。结果表明,长期施肥或秸秆可以促进红壤水稻土氮素的积累,其主要影响020cm土层,而化肥和秸秆配合施用作用更明显;长期施NPK化肥使土壤C/N降低,而秸秆还田使土壤C/N明显升高。在两种温度下的土壤矿质氮含量和累积矿化量曲线比较相似,10℃下土壤矿化速率达到稳定所需的时间要比30℃下长;30℃下不同施肥处理之间的矿化过程存在显著差异,而10℃下的差异较小,其在56d内矿化出来的矿质氮是10℃下的近两倍。化肥NP（仅施化肥NP）及NP+C（施化肥NP+秸秆还田）处理均有较高氮素矿化量,秸秆还田对土壤氮素矿化的促进作用比仅施化肥明显。在30℃下,土壤积累矿化量与有效积温的关系较好地符合有效积温方程式（EATM）（p0.01）。仅施化肥处理和对照方程中的K值相差不大,而秸秆还田与化肥配施处理方程的K值则有增大趋势,NPK+C（施化肥NPK+秸秆还田）处理的矿化潜力较大,且养分供应能力较强;秸秆配施化肥处理方程中n值的绝对值有减小趋势。随着培养时间延长,秸秆还田配施化肥处理土壤的累积矿化量保持在一个较稳定范围。     

Characterization of phytochemicals and antioxidant activities of a purple tomato (Solanum lycopersicum L.)

A newly developed nongenetically modified purple tomato V118 was investigated for its phytochemical compositions and antioxidant activities. A highly efficient and sensitive UPLC method was developed for both the phenolics and carotenoids, which showed that in addition to the phytochemicals commonly known for tomatoes, V118 had a unique composition of anthocyanins. The total carotenoid content of V118 was 234.78 μg/g dry weight (DW), and the total phenolic content was 659.11 mg GAE/100 g DW. The antioxidant activities of the lipophilic extract as measured by the PCL and ORAC-L assays were 30.11 μmol TE/g DW and 11.97 μmol TE/g DW, respectively, while the hydrophilic extracts as determined by the ORAC-H and FRAP assays were 323.23 μmol TE/g DW and 54.95 μmol AAE/g DW, respectively. The LC-MS study showed three major anthocyanins, which were mainly acylglycosides of petunidin and malvidin. This study showed that purple tomatoes such as V118 possess additional phytochemicals like anthocyanins, which can potentially have added health benefits.     

Novel royal jelly proteins identified by gel-based and gel-free proteomics

Royal jelly (RJ) plays an important role in caste determination of the honeybee; the genetically same female egg develops into either a queen or worker bee depending on the time and amount of RJ fed to the larvae. RJ also has numerous health-promoting properties for humans. Gel-based and gel-free proteomics approaches and high-performance liquid chromatography-chip quadruple time-of-flight tandem mass spectrometry were applied to comprehensively investigate the protein components of RJ. Overall, 37 and 22 nonredundant proteins were identified by one-dimensional gel electrophoresis and gel-free analysis, respectively, and 19 new proteins were found by these two proteomics approaches. Major royal jelly proteins (MRJPs) were identified as the principal protein components of RJ, and proteins related to carbohydrate metabolism such as glucose oxidase, α-glucosidase precursor, and glucose dehydrogenase were also successfully identified. Importantly, the 19 newly identified proteins were mainly classified into three functional categories: oxidation-reduction (ergic53 CG6822-PA isoform A isoform 1, Sec61 CG9539-PA, and ADP/ATP translocase), protein binding (regucalcin and translationally controlled tumor protein CG4800-PA isoform 1), and lipid transport (apolipophorin-III-like protein). These new findings not only significantly increase the RJ proteome coverage but also help to provide new knowledge of RJ for honeybee biology and potential use for human health promotion.     

Comparison of the chemical constituents of aged pu-erh tea, ripened pu-erh tea, and other teas using HPLC-DAD-ESI-MSn

Pu-erh tea is a popular beverage in southwestern China and South Asian countries. To explain the differences of aged pu-erh tea and ripened pu-erh tea, the chemical constituents of these teas were identified by HPLC-DAD-ESI-MS(n). In addition, HPLC was used to determine the contents of the major polyphenols, gallic acid, caffeine, and theobromine, in various types of teas. These results showed that the majority of chemical constituents in ripened pu-erh tea and aged pu-erh tea were similar, but the contents of catechins and gallic acid presented significant differences between these two teas. After fermentation by microorganism, the levels of catechins in ripened pu-erh tea were decreased, but the contents of gallic acid and caffeine were conversely elevated compared with aged pu-erh tea.     

8份广西产绞股蓝种质的RAPD引物筛选及其遗传多样性分析

本研究的目的在于筛选合适的RAPD随机引物,应用RAPD技术对药用植物绞股蓝进行遗传多样性分析,并构建DNA指纹图谱。研究结果表明,我们利用生物信息学方法挑选出的20条引物中有19条引物的扩增条带清晰且多态性好;在清晰稳定出现的354条带中,294条具有多态性;其中有3条引物的扩增条带可清楚区分绞股蓝与混淆品种乌蔹莓,可建立其DNA指纹图谱。按UPGMA法进行聚类分析,计算其遗传相似系数,结果显示,8份绞股蓝供试材料聚为两类,聚类结果与其地理区域远近和生长环境一致。本研究中筛选出的19条引物适用于绞股蓝遗传多样性分析,且获得的DNA指纹图谱可用于鉴别绞股蓝。     

均匀设计优化毛竹EST-SSR PCR反应体系

本研究以毛竹叶片DNA为模板,利用均匀设计U10（54）表,对毛竹EST-SSRPCR反应体系的TaqDNA聚合酶、Mg2＋、dNTPs及引物浓度4因素5个水平进行优化筛选。优化实验结果表明,毛竹EST-SSRPCR的25μL优化反应体系为：10×PCRBuffer（含Mg2＋）2.5μL、TaqDNA聚合酶1.5U,Mg2＋、dNTPs及引物的最适浓度分别是1.0mmol/L、0.1mmol/L、0.48μmol/L;通过梯度PCR试验筛选得到相应引物的最佳退火温度为52.4℃。对优化出的EST-SSRPCR反应体系进行稳定性检测,结果均能获得丰富、稳定、清晰可辨的DNA谱带。该优化体系为利用EST-SSR分子标记对毛竹进行遗传多样性等相关研究工作提供了基础条件。     

苹果杂交后代植株果实着色性状的遗传分析和预测

苹果着色问题是影响我国许多产区苹果质量的重要因素,本研究利用苹果着色相关基因MdMYB1（GenBank登录号：DQ886414）设计引物进行PCR扩增,开发出一个酶切位点为HaeⅢ的CAPS标记Mb2,该标记可用于区分杂交亲本‘富士’和‘嘎拉’苹果及分析其杂交后代植株。进一步利用Mb2标记对该杂交组合的后代植株进行遗传分析,结果表明,可将77个后代植株分为2组,一组42个后代植株含有来自‘嘎拉’亲本的非红色性状等位基因以及来自‘富士’亲本的非红色性状等位基因或者红色性状等位基因,其果实颜色预测为非红色与红色或者非红色与桔红色;另一组35个后代植株含有来自‘嘎拉’亲本的红色性状等位基因以及来自‘富士’亲本的非红色性状等位基因或者红色性状等位基因,其果实颜色预测为桔红色与红色或者只有红色。本研究结果将为选育优质苹果新品种提供依据和方法参考。