雷公藤鲨烯环氧酶基因克隆与表达分析

以雷公藤(Tripterygium wilfordii Hook.f.)发状根为试验材料,采用RT-PCR方法,克隆得到2个雷公藤鲨烯环氧酶编码基因,命名为TwSE1(GenBank登录号MG717395)和TwSE2(GenBank登录号MG717396)。序列分析表明,TwSE1和TwSE2的开放阅读框分别为1 578和1 584bp,分别编码525和527个氨基酸,2条序列相似性为76.18%,但N端序列不保守。实时荧光定量PCR检测雷公藤鲨烯环氧酶在不同组织部位的表达模式,以及雷公藤发状根受茉莉酸甲酯(MeJA)诱导后基因表达的结果表明,TwSE1、TwSE2基因在雷公藤根、茎、嫩叶、老叶、花中均有表达,TwSE1在花中表达丰度最高,在根中表达丰度最低;但TwSE2在花和嫩叶中表达量最高,在老叶中表达量最低,且TwSE2在各组织部位的表达量都低于TwSE1。雷公藤发状根经MeJA诱导后,TwSE1、TwSE2基因表达量均上升,都表现为表达量先上升后下降再上升的趋势,并于诱导后3h达到一个高水平表达,之后下降,在诱导12h后表达量又迅速上升;在相同诱导条件下,TwSE2基因表达水平的提高小于TwSE1基因。     

Comparative proteomic analysis reveals the roots response to low root-zone temperature in <Emphasis Type="Italic">Malus baccata</Emphasis>

Soil temperature is known to affect plant growth and productivity. In this study we found that low root-zone temperature (LRT) inhibited the growth of apple (Malus baccata Borkh.) seedlings. To elucidate the molecular mechanism of LRT response, we performed comparative proteome analysis of the apple roots under LRT for 6 days. Total proteins of roots were extracted and separated by two-dimensional gel electrophoresis (2-DE) and 29 differentially accumulated proteins were successfully identified by MALDI-TOF/TOF mass spectrometry. They were involved in protein transport/processing/degradation (21%), glycometabolism (20%), response to stress (14%), oxidoreductase activity (14%), protein binding (7%), RNA metabolism (7%), amino acid biosynthesis (3%) and others (14%). The results revealed that LRT inhibited glycometabolism and RNA metabolism. The up-regulated proteins which were associated with oxidoreductase activity, protein metabolism and defense response, might be involved in protection mechanisms against LRT stress in the apple seedlings. Subsequently, 8 proteins were selected for the mRNA quantification analysis, and we found 6 of them were consistently regulated between protein and mRNA levels. In addition, the enzyme activities in ascorbate–glutathione (AsA–GSH) cycle were determined, and APX activity was increased and GR activity was decreased under LRT, in consistent with the protein levels. This study provides new insights into the molecular mechanisms of M. baccata in responding to LRT.     

水牛卵巢黄体形成和退化的蛋白质表达谱构建及生物信息学分析

本研究构建了水牛卵巢黄体形成及退化过程的定量蛋白质表达谱,从中寻找与水牛发情周期相关的重要蛋白质。为研究水牛卵巢周期性变化的分子机制提供蛋白质水平的数据支撑,进而为提高水牛繁殖效率奠定实验基础。以水牛排卵后的红体期(corpus hemorrhagicum,CH)、黄体期(corpus luteum,CL)和白体期(corpus fibrosum,CF)的卵巢作为研究对象,利用串联质谱标签(tandem mass tag,TMT)标记结合液质联用((liquid chromatography/mass spectrometry,LC-MS/MS)的定量蛋白质组学技术,采用相关软件对得到的差异表达蛋白质进行生物信息学分析。同时对与水牛发情周期相关的重要差异蛋白质:单核巨噬细胞分化抗原(CD14)、通用转录因子Ⅱ-Ⅰ(GTF2I)、羟基类固醇(17β)脱氢酶1(HSD17B1)、U6 sn RNA相关Sm样蛋白质(LSM1)和纤溶酶原激活物抑制物(SERPINE1)进行了qRT-PCR分析验证。结果显示,共鉴定得到2 343种蛋白质,其中差异表达蛋白质有284种(差异倍数≥2.0),初步构建了水牛卵巢黄体形成和退化的定量蛋白质表达谱。对其中的五种重要差异蛋白质(CD14、GTF2I、HSD17B1、LSM1和SERPINE1)进行qRT-PCR验证,结果有四种(CD14、HSD17B1、LSM1和SERPINE1)的qPCR结果与质谱结果相一致,仅GTF2I与质谱结果不一致,可能是由于GTF2I存在转录后调控。本研究首次从蛋白质水平对水牛卵巢排卵后的周期性变化的分子机制进行了探究,同时为其他家畜品种发情周期的研究提供了新的研究思路。     

大熊猫DNA序列变异及其遗传多样性研究

测定了来自马边、美姑、越西、宝兴、平武、青川、南坪和白水江等地40只大熊猫336～444bp的线粒体tRNA基因和D环区序列.在21个创立者中共检出9种线粒体DNA单倍型.发现大熊猫的遗传分化程度很低,地理群体间无显著的遗传隔离.大熊猫现生群体可能源于晚更新世,并受瓶颈效应的影响.在此之后,其遗传多样性得到了一定程度的恢复.     

黄瓜花叶病毒坏死型卫星RNA的cDNA克隆和全序列分析

从田间发生坏死病害的番茄病株中分离得到黄瓜花叶病毒TN分离物,经研究证实TN分离物带有1株坏死型卫星RNA(TN-SatRNA).利用已知CMV卫星RNA的两端序列作引物扩增并克隆了TN-SatRNA的cDNA.序列分析显示TN-SatRNA全长为390个核苷酸.比较TN-SatRNA与富有代表性的CMV卫星RNA的结构表明,这几种卫星RNA中具有4个结构同源区(Ⅰ:1～81nt,Ⅱ:216～261nt,Ⅲ:278～338nt;Ⅳ:349～390nt),而在82～215nt的区域几种卫星RNA的结构变化较大.在TN-SatRNA的3’端具有已报道的坏死型卫星RNA特征性序列结构.经3’RACE方法确定TN-SatRNA的3’端自然序列,与PCR扩增引物有1个核苷酸的差别.     

野薄荷叶腺毛的发育形态学研究

研究了分布于我国东北部的野薄荷叶表面腺毛的结构和发育。主要腺毛有两种：多细胞盾状腺毛和弯曲单细胞头状腺毛。多细胞盾状腺毛由１个基细胞,１个柄细胞和一个由８－１２个分泌细胞构成的头部组成。单细胞头状腺毛由１个基细胞,１个柄细胞和一个分泌细胞头部构成。     

甘蔗螟虫卵赤眼蜂繁殖利用的研究

甘蔗是我国最重要的糖料作物,甘蔗害虫以甘蔗螟虫最普遍,分怖亦最廣。廣东珠江三角洲常见的甘蔗螟虫共有四种:条螟或称斑点螟(Diatraea venosata Wk.)、二点螟(Chilo infuscatellus Snellen)、黄螟(Eucosma schistaceana Snellen)和大螟(Sesamiainferens Wk.)。甘蔗螟虫为害的结果,形成枯心苗和蛀(?),影响甘蔗生长发育,减低蔗糖成份,易受风折,而且造成甘蔗赤腐病菌入侵条件。 甘蔗螟虫的为害虽重,但目前还没有一套完整的防治方法,也还没有一种很有效的方法。本试验目的,是利用赤眼蜂来防治甘蔗螟虫。试验内容是赤眼蜂的寄主的选择和繁殖、赤眼蜂的繁殖及保存、赤眼蜂田间放播初步试验和甘蔗螟虫田间发生情况的调查。现将各项试验结果简要地分述如下: 赤眼蜂能否利用成功,要看培育出来的赤眼蜂是否具有高度的生活力。赤眼蜂的生活力可用下列四个标准去量度:(1)蜂体大小,(2)繁殖能力,(3)成虫寿命,(4)对田间环境的適应性。此外,繁殖出来的赤眼蜂雌性此率不应此自然界的减低。 赤眼蜂的生活力,首先和寄主有很大关系。如果寄主卵的内含物的质和量都適合 於赤眼蜂幼虫营养之需,羽化出来的成虫体积大,繁殖力强,寿命也有延长的趣势。关於赤眼蜂的寄主,我们管用过17种鳞翅目昆虫的卵供其寄生,结果以松毛虫(Dendro     

高雄山虫草及其细脚拟青霉无性型

报道采自安徽省天堂寨自然保护区的高雄山虫草Cordyceps takaomontana及其无性型细脚拟青霉Paecilomyces tenuipes,应用单子囊孢子分离鉴定和微循环产孢的方法确证了两者的对应关系,并修订了高雄山虫草的原始描述。双梭孢虫草C．Bifusispora可能为高雄山虫草的同物异名。同时采集的具数个红色子座且有双梭形子囊孢子的虫草不是同一个种。     

Effect of Elevated CO2 on the Primary Conversion of Light Energy of Alfalfa Photosynthesis

In studying the mechanism of increase in alfalfa (Medicago sativa L. ) photosynthesis under elevated atmospheric CO2, it was found that the capacity of chloroplasts for light absorption was greater, the potential activity and efficiency of primary conversion of light enlergy of PS Ⅱ , quantum yield of PS Ⅱ electron transport, and activation capacity of PS Ⅰ were stimulated, photochemical quenching coefficient was increased and non-photochemical quenching coefficient was decreased under elevated atmospheric CO2.