8个山葡萄及山欧杂种葡萄品种的SSR分析

选取9对SSR引物对8个山葡萄及山欧杂种葡萄品种进行扩增,估算了扩增片段长度,结果表明,利用这9对SSR引物可以把8个山葡萄及山欧杂种葡萄品种区分.8个山葡萄及山欧杂种葡萄品种的等位基因数(Na)平均为2.8889,有效等位基因数(Ne)平均为2.2162,Shannon多样性指数(I)平均为0.8669,期望杂合度(Nei)平均为0.5217.9对引物中,UDV-067提供的信息量最大,引物VMC7b1最小.     

Establishment of a multi-color genomic in situ hybridization technique to simultaneously discriminate the three interspecific hybrid genomes in gossypium

To identify alien chromosomes in recipient progenies and to analyze genome components in polyploidy, a genomic in situ hybridization （GISH） technique that is suitable for cotton was developed using increased stringency conditions. The increased stringency conditions were a combination of the four factors in the following optimized state： 100：1 ratio of blocking DNA to probe, 60% formamide wash solution, 43 ℃ temperature wash and a 13 min wash. Under these specific conditions using gDNA from Gossypium sturtianum （C1 C1 ） as a probe, strong hybridization signals were only observed on chromosomes from the C1 genome in somatic cells of the hybrid F1 （G. hirsutum x G. sturtianum） （AtDtC1）. Therefore, GISH was able to discriminate parental chromosomes in the hybrid. Further, we developed a multi-color GISH to simultaneously discriminate the three genomes of the above hybrid. The results repeatedly displayed the three genomes, At, Dt, and C1, and each set of chromosomes with a unique color, making them easy to identify. The power of the multi-color GISH was proven by analysis of the hexaploid hybrid F1 （G. hirsutum x G. australe） （AtAtDtDtG2G2）. We believe that the powerful multi-color GISH technique could be applied extensively to analyze the genome component in polyploidy and to identify alien chromosomes in the recipient progenies.     

Lead induced changes in the growth and antioxidant metabolism of the lead accumulating and non-accumulating ecotypes of Sedum alfredii

The phytotoxicity and antioxidative adaptations of lead （Pb） accumulating ecotype （AE） and non-accumulating ecotype （NAE） of Sedum alfredii Hance were investigated under different Pb treatments involving 0, 0.02 mmol/L Pb, 0.1 mmol/L Pb and 0.1 mmol/L Pb/0.1 mmol/L ethylenediaminetetraacetic acid （EDTA） for 6days. With the increasing Pb level, the Pb concentration in the shoots of AE plants enhanced accordingly, and EDTA supply helped 51% of Pb translocation to shoots of AE compared with those treated with 0.1 mmol/L Pb alone. Moreover, the presence of EDTA alleviated Pb phytotoxicity through changes in plant biomass, root morphology and chlorophyll contents. Lead toxicity induced hydrogen peroxide （H2O2） accumulation and lipid peroxidation in both ecotypes of S. alfredii. The activities of superoxide dismutase （SOD）, guaiacol peroxidase （G-POD）, ascorbate peroxidase, and dehydroascorbate reductase elevated in both leaves and roots of AE as well as in leaves of NAE with the increasing Pb levels, but SOD and G-POD declined in roots of NAE. Enhancement in glutathione reductase activity was only detected in roots of NAE while a depression in catalase activity was recorded in the leaves of NAE. A significant enhancement in glutathione and ascorbic acid （AsA） levels occurred in both ecotypes exposed to Pb and Pb/EDTA treatment compared with the control, however, the differences between these two treatments were insignificant. The dehydroascorbate （DHA） contents in roots of both ecotypes were 1.41 to 11.22-fold higher than those in leaves, whereas the ratios of AsA to DHA （1.38 to 6.84） in leaves altering more to the reduced AsA form were much higher than those in roots. These results suggested that antioxidative enzymes and antioxidants play an important role in counteracting Pb stress in S. alfredii.     

动脉粥样硬化大鼠动脉的基因芯片分析及组织蛋白酶D的表达变化

目的:用基因芯片技术研究动脉粥样硬化大鼠与正常大鼠动脉组织中差异表达的基因,以探讨动脉粥样硬化的发病机制.方法:16只雄性SD大鼠随机分组,正常对照组8只给予正常饮食及生理盐水腹腔注射,模型组8只大鼠喂食高脂饲料(3%胆固醇、0.5%胆酸钠、5%精制糖、10%猪油、基础饲料81.5%)及维生素D3腹腔注射,实验14周处死动物,测定血浆总胆固醇(total cholesterol,TC)及甘油三酯(triglyeride,TG),取主动脉进行HE染色及油红染色进行病理学观察,提取主动脉RNA作基因芯片检查,以免疫组织化学方法检测组织蛋白酶D(Cathepsin D,Cat-D)蛋白表达情况.结果:与正常对照组相比.模型组血浆TG和TC升高(P<0.01),HE及油红染色可见主动脉形成纤维增生性动脉粥样硬化.芯片结果显示,与空白对照组相比.模型组Cat-D基因表达上调.进一步免疫组化检查发现,正常对照组大鼠动脉组织中Cat.D不表达或者低表达,模型组中Cat-D表达明显增加(P<0.0).结论:AS的发病是多因素作用的多基因改变而引发的复杂病变,组织蛋白酶D可能与其他具有降解基质作用的酶协同参与了血管重塑从而促进AS的发展.     

大气CO2浓度升高对八宝景天生长及光合生理的影响

利用开顶式气室(OTC)系统,设正常大气CO₂浓度和CO₂浓度升高200 μmol·mol^-12个CO₂浓度处理,模拟大气CO₂浓度升高对八宝景天光合生理和生长发育的影响.结果表明: 大气CO₂浓度升高使八宝景天叶片上、下表皮气孔密度分别显著下降16.1%和16.7%,使叶片维管束增粗,导管增多,靠近上表皮细胞增大;CO₂浓度升高可以显著增加傍晚时八宝景天叶片光合色素含量,使夜间净光合速率、气孔导度和蒸腾速率显著增加.初花期傍晚,CO₂浓度升高使叶片苹果酸含量显著下降64.0%,纤维素含量显著增加20.8%.盛花期清晨,CO₂浓度升高使叶片苹果酸含量显著增加27.0%,对糖类物质含量影响不显著,植株的分枝数、单株茎质量和单株总生物量显著增加.CO₂浓度升高可以促进八宝景天光合作用,有利于植株生长.     

黄原胶发酵培养基优化研究

分别用单因素法和正交试验法对野油菜黄单胞菌J-12的发酵培养基成分进行研究。得到的最优培养基配方为:4％玉米淀粉、0.3％鱼粉、0.3％豆饼粉、0.3％CaCO_3、0.5％KH_2PO_4、0.25％MgSO_4、0.025％FeSO_4、0.025％柠檬酸,接种量5％。在消前pH7.2～7.5,发酵温度28℃,摇床转速180r/min,发酵时间72h的实验条件下,发酵液粘度为8740mPa·s,采用酒精直接沉淀法提取黄原胶的产率达2.91％,产品的丙酮酸含量3.32％。     

镉在东南景天中的亚细胞分配

采用差速离心技术,比较研究了Cd在矿山生态型东南景天(Sedum alfredii Hance)根、茎、叶中的亚细胞分布。结果表明:矿山生态型东南景天对Cd具有很好的忍受耐和累积能力,非矿山生态型东南景天则不具有这种能力。Cd在矿山生态型东南景天根、茎、叶各部分的亚细胞分配满足F1(细胞壁部分)>F3(可溶部分)>F2(细胞器及膜)规律,且Cd在细胞壁部分的分配占绝对优势;同时矿山生态型东南景天地上部有很好的Cd累积能力。与众多超累积植物相似的Cd亚细胞分配规律和地上部的良好的Cd累积能力。对此植物的Cd亚细胞分布完全不同于非矿山生态型东南景天的情况作了比较分析,结果显示矿山生态型东南景天很可能是一种新的镉超累积种质资源。     

古老铅锌矿区生态型东南景天对锌耐性及超积累特性的研究

 植物长期生长在重金属污染的生境中,逐渐进化成不同的生态型。通过调查中国东南部古老Pb/zn矿和非矿山生境中的植物种群,发现生长在古老Pb／Zn矿的东南景天(Sedum alfredii Hance)是一种新的Zn超积累植物。在自然和控制条件下,古老Pb／Zn矿生态型比非矿山生态型植株的茎粗、叶片大、植株高。在矿山土壤Zn有效含量为105.5～325.4mg·kg-1时,矿山生态型东南景天植株地上部Zn含量为4134～5000mg·kg-1;当营养液中Zn浓度为1223.6μmol时,其Zn含量高达2％。在相同Zn浓度下,矿山生态型地上部Zn含量比非矿山生态型高30倍左右。两种生态型体内Zn分布也不同,古老铅锌矿山生态型的不同器官中Zn含量以茎>叶片>根系,而非矿山生态型则以根系>茎>叶片。古老铅锌矿山生态型地上部积累的Zn占植株总积累量的90％以上,其中叶片和茎分别占41.66％±5.46％和54.75％±5.87％;非矿山生态型各器官中积累的Zn远远低于古老铅锌矿山生态型,各器官中积累的Zn以茎>根系>叶片。本研究表明,这两种生态型东南景天的发现,为今后探讨植物耐高Zn胁迫和超积累Zn的微进化过程提供了非常有价值的材料,也为Zn污染土壤的植物修复提供了一种很有潜力的候选材料。     

小麦细胞核仁中DNA原位位置与rRNA基因转录位点的研究

以小麦细胞为研究材料, 应用常规电子显微镜技术和DNA细胞化学特异染色NAMA-Ur技术, 在原位水平对核仁中DNA的分布和特征进行了直观的观察。结果表明, 小麦细胞核仁中DNA位于纤维中心(Fibrillar Centers, FC)、致密纤维组分(Dense Fibrillar Component, DFC)以及两者的过渡区域, 并呈现出环绕FC排布的构型; 应用RNP优先染色(Benhard staining)技术分析了核仁中RNP的分布及其原位位置, 直观的显示了小麦细胞核仁中RNP颗粒主要集中在 FC与DFC的过渡区域及DFC和颗粒组分(Granular Component, GC)中; 并且在FC与DFC的过渡区域, 它们不太均匀也不太连续地半围绕着FC而排布; 进一步借助于RNA/DNA杂合体抗体在原位水平标记和分析了细胞核仁中活跃基因转录的精细位点, 结果表明小麦细胞核仁rRNA基因的转录位点位于FC与DFC的过渡区域及DFC中。     

栗的 PGI遗传和多样性

采用超薄聚丙烯酰４胺平板凝胶等电聚焦电泳和单株后代法,分析了栗属种的ＰＧＩ同工酶的遗传,研究发现Ｐｇｉ位点（Ｐｇｉ－ｉ）主要有３个等位基因并呈共显性遗传。在栗属的自然居群中还检测了出现频率较少的另外２个等位基因。