基于转录组的苹小卷叶蛾杀虫剂靶标及解毒代谢相关基因分析

【目的】建立苹小卷叶蛾Adoxophyes orana转录组数据库,挖掘杀虫剂靶标及解毒代谢相关基因。【方法】采用Illumina HiSeq~(TM) 2000高通量测序技术对苹小卷叶蛾进行转录组测序,挖掘并分析杀虫剂靶标基因;利用qPCR检测6个杀虫剂靶标基因在苹小卷叶蛾卵、幼虫、蛹和成虫各不同发育阶段的表达;挖掘并分析苹小卷叶蛾转录组中解毒代谢相关基因的代谢通路及进化关系。【结果】通过组装有效序列共获得48 610条unigene(GenBank登录号:GGMW00000000)。挖掘鉴定到155个杀虫剂靶标unigene;qPCR结果显示,1个蜕皮激素受体(ecdysone receptor, ECR)、2个乙酰胆碱酯酶(acetylcholinesterase, AChE)、1个氯离子通道蛋白(chloride channel, CLC)、1个几丁质酶(chitinase, CS)和1个鱼尼丁受体(ryanodine receptor, RyR)基因在苹小卷叶蛾不同发育阶段均存在表达差异。挖掘鉴定到69个羧酸酯酶(carboxylesterase, CarE)unigene、66个谷胱甘肽S-转移酶(glutathion S-transferase, GST)unigene和205个细胞色素P450(cytochrome P450)unigene等解毒代谢相关基因,共鉴定20个CarE unigene, 32个GST unigene和30个P450 unigene与有毒物质代谢相关的通路有关。基于氨基酸序列对具有完整ORF的unigene聚类分析结果显示:12个CarEs中9个为G类,即鳞翅目保幼激素类;分别有10个AoGSTs属于Delta和Epsilon亚家族;18个P450全部聚到CYP3集团。【结论】该研究有助于苹小卷叶蛾杀虫剂靶标基因的挖掘及抗药性的研究。     

树鼩IL-2全长编码序列的克隆及分子特征分析

树鼩作为多种人类疾病模型已受到广泛关注,而免疫因子对于树鼩模型评价至关重要,但目前对其白细胞介素-2(IL-2)的研究鲜有报道。该实验以经ConA(concanavalin)诱导培养的树鼩淋巴细胞总RNA为模板,RT-PCR克隆出465bp的树鼩IL-2全长编码序列,并采用ClustalW软件分析其序列和分子特征。结果表明树鼩IL-2cDNA编码一个由154个氨基酸组成的蛋白质,其cDNA及氨基酸序列与人的同源性分别为93%及80%,且其整体结构与人IL-2相似。MEGA5.0软件构建的进化树表明,树鼩与人及恒河猴的亲缘关系较近。Pymol软件对树鼩和人IL-2氨基酸序列进行的三维结构模建表明,两者的IL-2分子三维空间结构基本相似,表面大部分区域所带电荷相同,但在某些区域差异较大,且树鼩多出一个糖基化位点,这些差异对抗体的结合可能存在影响。该研究为今后树鼩IL-2单克隆抗体的制备及功能研究奠定了基础。     

‘香槟’月季的组织培养和试管开花诱导

本文对‘香槟’月季（80sachinensis‘Xiangbin’）的组织培养技术和诱导试管开花进行了研究。结果表明：以茎段为外植体能诱导获得无菌苗,适宜的启动培养基为MS＋6-BA1．0mg-L-1＋IBA0．1mg·L-1,幼芽继代增殖的最佳培养基是MS＋6．BA1．0mg·L-1。＋IBA0．1～0．2mg·L-1,诱导生根的适宜培养基为1／2MS＋NAA0．3mg·L-1,生根率达80．0％。诱导试管开花的适宜培养基为MS＋6．BA0．5mg·L-1＋NAA0．1mg·L-1最适宜的诱导试管开花的蔗糖含量是30g·L-1;在三角瓶中培养,试管花可以正常开放,在培养瓶中培养花芽不能正常开放;MS培养基中增加2倍磷的含量,可以提高花芽诱导率,为25．O％;诱导试管开花的最适培养条件为温度21℃,光照强度80～100μmol·m-2．s-1,光照时间16h—d-1。     

Evaluation of the larvicide effect of an aqueous extract of Persea americana (Miller, 1768), Lauraceae,on Anopheles gambine (Giles, 1902), Diptera Culicidae

Summary

In-laboratory evaluation of the larvicide effect of an aqueous extract of Persea americana (Miller 1768) on the various larval stages of Anopheles gambiae (Giles 1902) has shown that larvae at all stages are sensitive to this extract. Inversely proportional to the larval stage, their sensitivity proved to depend on the level of concentration: the higher the latter, the higher the former. Indeed, after a 24-h exposure, concentrations of 510 μg/ml resulted in the death of 100% of larvae at stages 1 and 2, whereas for the same time of exposure there was need to resort to concentrations of 850 μg/ml to kill them at stages 3 and 4. The aqueous extract of Persea americana has therefore a poisonous effect on larvae of Anopheles gambiae. This extract has direct lethal effects and acts as an inhibitor of larval development at sublethal concentration levels.     

Overexpression of the phosphatidylinositol synthase gene (ZmPIS) conferring drought stress tolerance by altering membrane lipid composition and increasing ABA synthesis in maize

Phosphatidylinositol (PtdIns) synthase is a key enzyme in the phospholipid pathway and catalyses the formation of PtdIns. PtdIns is not only a structural component of cell membranes, but also the precursor of the phospholipid signal molecules that regulate plant response to environment stresses. Here, we obtained transgenic maize constitutively overexpressing or underexpressing PIS from maize (ZmPIS) under the control of a maize ubiquitin promoter. Transgenic plants were confirmed by PCR, Southern blotting analysis and real‐time RT‐PCR assay. The electrospray ionization tandem mass spectrometry (ESI‐MS/MS)‐based lipid profiling analysis showed that, under drought stress conditions, the overexpression of ZmPIS in maize resulted in significantly elevated levels of most phospholipids and galactolipids in leaves compared with those in wild type (WT). At the same time, the expression of some genes involved in the phospholipid metabolism pathway and the abscisic acid (ABA) biosynthesis pathway including ZmPLC, ZmPLD, ZmDGK1, ZmDGK3, ZmPIP5K9, ZmABA1, ZmNCED, ZmAAO1, ZmAAO2 and ZmSCA1 was markedly up‐regulated in the overexpression lines after drought stress. Consistent with these results, the drought stress tolerance of the ZmPIS sense transgenic plants was enhanced significantly at the pre‐flowering stages compared with WT maize plants. These results imply that ZmPIS regulates the plant response to drought stress through altering membrane lipid composition and increasing ABA synthesis in maize.     

从烟草EST微卫星标记中筛选合子胚中优势表达的基因

为获得烟草合子胚中的优势表达基因,利用CAP3程序对来自烟草公共数据库的EST序列进行组装,利用MISA程序从组装后的EST中筛选SSR位点,将多态性的SSR位点在烟草合子文库中进行扩增并对等位基因进行分析。结果表明:具有多态性的16个SSR标记中,有9个基因能从烟草的合子库中成功扩增得到。该研究为筛选烟草合子胚中优势表达基因提供新的途径。     

二郎山小型兽类区系及分布格局

动物种类和数量在不同的山地或同一山地的不同垂直高度有很大的差异,正因如此,国际上和国内的研究者们一直在努力探讨动物在山地的分布规律和产生差异的原因(Merriam et al.,1890;鲍毅新等,1984;Heaney,2001;Rickart,2001;SánchezCordero,2001;张云智等,2002;李义明等,2003;马俊等,2010).横断山是中国最长、最宽和最典型的南北向山系,位于青藏高原东南部,通常为四川、云南两省西部和西藏自治区东部南北向山脉的总称.横断山是全球25个生物多样性热点地区之一,其生物多样性资源极其丰富(Myers et al.,2000).     

广西柿种质资源果实性状多样性分析与模糊综合评价

为进一步揭示广西柿种质资源信息,运用多样性分析和模糊综合评价法对不同生态气候区的柿种质资源果实性状进行研究和评价。结果表明:不同柿种质的果实性状存在丰富的多样性。86份柿种质的模糊综合评价指数FCI值在1.849～3.947之间,平均2.835。根据FCI取值范围将所调查的柿种质分成5个等级,FCI值最高的第5级种质数量最少,占所调查种质的3.49%;FCI值最低的第1级种质数量最多,占所调查种质的48.84%。月柿、牛心柿、大方柿、京柿、鸡心柿、小方柿和火柿等柿品种及一些实生单株的模糊综合评价指数较高,它们的果实综合性状表现较优,建议作为优良品种和单株加以开发和利用。部分单个性状比较优良的实生单株虽不适合于直接推广种植,但可作为柿品种改良的资源加以利用。     

苎麻属野生植物纤维细胞形态结构与其经济性状和物理性能的关系

以13种苎麻属植物为试验材料,于植物工艺成熟期取样测定经济性状并用显微镜观察横截面及离析纤维细胞形态,研究苎麻属植物纤维细胞形态结构与其经济性状和物理性能的相关性。结果显示:(1)苎麻属野生种质的纤维细胞外观形态表现与栽培种苎麻基本一致,多呈圆形、多边形、椭圆形和肾形等,形态各异,大小不一。(2)茎秆横截面纤维细胞直径、腔径和纤维胞总数都明显少于对照栽培种,与其原麻产量存在不同程度的正相关性。(3)离析纤维细胞长度、宽度、壁厚和结节数与纤维细度存在不显著负相关,且野生苎麻属种质的细胞长度、宽度明显小于栽培种苎麻。研究表明,野生苎麻属种质在提高栽培种产量上无太大效应,但其在优良纤维基因选育方面和改良栽培种苎麻纤维品质上具有重要应用价值。