白菜型油菜八氢番茄红素脱氢酶基因PDS3的克隆与序列分析

利用电子克隆技术,从数据库中找到与拟南芥高度同源的油菜EST序列,继而通过RT-PCR和RACE等方法,成功克隆了白菜型油菜八氢番茄红素脱氢酶（类胡萝卜素合成途径中的一个关键限速酶）基因PDS3的cDNA,命名为BrPDS3（GenBank 登记号GQ200741）。cDNA序列全长1940bp,其中包含114bp的5’前导序列和134bp的3’不翻译序列,编码区长度为1 692bp,编码63kD的蛋白。序列分析表明,BrPDS3蛋白与其他植物的PDS蛋白具有较高相似性;在系统进化树中,BrPDS3与甘蓝亲缘关系最近。根据全长cDNA序列设计引物,从白菜型油菜青油13号DNA中克隆得到BrPDS3基因的全长DNA,长度为3 911bp,ORF（开放阅读框）1 692bp,含有15个外显子和14个内含子。     

甘蓝型油菜BnPex16p基因cDNA的克隆与表达

利用模式植物拟南芥AtPex16p/sse1基因序列与油菜数据库BBSRC BrassicaDB比对,得到甘蓝型油菜(Brassica napus)同源EST序列,拼接出油菜Pex16p基因全长cDNA电子克隆,然后设计全长引物,以甘蓝型油菜种子cDNA为模板,克隆Pex16p基因,得到全长为1 101bp的cDNA,编码366个氨基酸的蛋白,命名为BnPex16p。BnPex16p基因序列与拟南芥AtPex16p/sse1同源,其蛋白与拟南芥同源蛋白具有完全相同的PTS2型过氧化物酶体定位肽,进化关系相近。半定量PCR(RT-PCR)发现BnPex16p基因在油菜幼嫩的根、茎、叶和种子中均有高丰度表达,在种子发育过程的中期和后期表达水平增加,暗示该基因在油脂的积累中有重要作用。     

大豆脱氢抗坏血酸还原酶基因的电子克隆及进化分析

利用生物信息学和实验验证的技术路线,以拟南芥脱氢抗坏血酸还原酶基因cDNA序列为信息探针,对大豆EST数据库进行同源搜索,经同源性比对和序列组装,获得全长为955bp的大豆脱氢抗坏血酸还原酶基因的cDNA序列(GenBank登录号为DQ006810),经RT-PCR扩增,分子克隆和序列分析验证,结果表明与电子克隆完全一致.该cDNA序列具有完整的开放阅读框架(ORF,36bp-815bp),推测编码蛋白为259个氨基酸.通过与几种植物脱氢抗坏血酸还原酶蛋白序列的比较,发现该基因具有较高的保守性.结果表明根据物种问同源基因序列,对跨物种问EST数据库进行同源检索筛选,拼接,是新基因克隆的一条有效途径.     

花生中两个硬脂酰-ACP去饱和酶基因的克隆与序列分析

脂肪酸去饱和酶是不饱和脂肪酸合成途径的关键酶,催化脂肪酸链特定位置上脱氢形成双键.本研究通过构建花生种子全长cDNA文库,结合大规模EST测序和RACE克隆等方法,从花生中克隆得到了两个FAB2基因,分别命名为AhFAB2-2和AhFAB2-3.其中AhFAB2-2全长为1387bp,ORF为1158bp,理论分子量为43.7 kD,理论等电点为5.69;AhFAB2-3全长为1452bp,ORF为1188bp,理论分子量为44.9 kD,理论等电点为6.37.它们推导的氨基酸序列与拟南芥的相似性依次为60.8％和57.2％;与大豆的相似性分别是62.3％和59.3％.这两个基因在GenBank上的登录号分别为KF358459和KF358460.     

茶树八氢番茄红素脱氢酶cDNA全长克隆与表达分析

八氢番茄红素脱氢酶是类胡萝卜素生物合成途径的关键酶之一。本实验采用3’/5’RACE和RTPCR技术成功扩增出茶树八氢番茄红素脱氢酶基因（PDS）的3’端和5’端序列,序列拼接获得全长cDNA序列,命名为CsPDS,并将其登录至GenBank,登录号KF646537。经生物信息学分析,所得基因cDNA序列全长为2295 bp,开放阅读框（ORF）1749 bp,编码582个氨基酸,预测分子量约为64.86 kDa,理论等电点（PI）为6.77,属于亲水性蛋白。该基因编码的氨基酸序列与柿树PDS序列的同源性达到87%,多序列比对表明茶树PDS具有高度保守区域,基于邻接法的进化树显示与柿树、葡萄的亲缘关系最近。实时荧光定量PCR分析结果显示,CsPDS在‘黄金芽’体内表达没有受抑制,表明‘黄金芽’黄色白化可能不是在CsPDS基因转录水平异常而引起。     

甘蓝型油菜BAN同源基因片段克隆与序列分析

参照拟南芥（Arabidopsis thaliana)BAN基因和cDNA的保守序列设计引物，对甘蓝型、白菜型、芥菜型油菜和拟南芥及其它十字花科栽培品种的基因组总DNA进行PCR扩增，均获得与拟南芥BAN基因扩增片段大小极其相似的PCR扩增产物，表明BAN基因可能广泛存在于十字花科植物中。甘蓝型油菜和拟南芥BAN扩增片段测序比较分析显示，拟南芥BAN片段（779bp)与已往的报道完全相同，甘蓝型油菜的BAN片段（780bp)与拟南芥BAN外显子的同源性为90％，但与内含子的同源性仅为74％。甘蓝型油菜BAN氨基酸序列与80多种植物的NADPH还原酶类有不同程度的同源性。     

巴西橡胶树HbMYC1基因的克隆及序列分析

从GenBank中搜索到1条来自橡胶树的MYC(Myelocytomatosis)家族基因EST序列,以该序列为保守区,通过RACE技术获得了该基因全长cDNA序列,共1817bp。通过BLAST工具搜索GenBank数据库,结果表明,该cDNA序列同其他植物的MYC家族基因高度同源,认为该cDNA序列为橡胶树MYC家族成员。利用GenScan和Clastal X对该序列进行分析,推测其编码区长度为1428bp,编码476个氨基酸。生物信息学分析结果表明,巴西橡胶树的MYC基因编码的氨基酸序列与其他物种的相似性很高,同时推导的蛋白质序列中还存在着1个螺旋-环-螺旋的DNA结合区域和1个核定位信号,表明克隆到的基因是1个MYC家族成员的转录因子。     

三个花生溶血磷脂酸酰基转移酶基因的克隆与序列分析

溶血磷脂酸酰基转移酶在植物油脂合成途径中具有重要的作用.本研究通过构建花生种子全长cDNA文库,结合大规模EST测序和RACE克隆等方法,从花生中克隆得到了三个LPAT基因,分别命名为AhLPAT2,AhLPAT4和AhLPAT5.其中AhLPA72全长为1626bp,ORF为1164bp,理论分子量为43.2 kD,理论等电点为9.42；AhLPAT4全长为1618bp,ORF为1152bp,理论分子量为43.9 kD,理论等电点为9.23；AhLPA T5全长为1911bp,ORF为1137bp,理论分子量为43.7 kD,理论等电点为8.99.它们推导的氨基酸序列与拟南芥的同源性依次为78％,65％和65％；与大豆的同源性分别是82％,80％和82％.将这三个基因在GenBank上注册,注册号分别为JN032677,KC160499,KC160500.     

甘蓝型油菜裂角相关基因BnRPL的克隆与分析

为鉴定甘蓝型油菜裂角相关基因，基于候选基因途径，利用同源克隆结合RACE方法，在甘蓝型油菜易裂角品系R2中扩增出了两个RPL基因的全长cDNA序列，其中一条序列长度为2 068bp，开放阅读框序列位置为154-1 911位，总长为1 758bp，命名为BnRPL.a。另一条序列全长为2 041bp，开放阅读框序列位置为154-1 887位，总长为1 734bp，命名为BnRPL.c。分析表明利用PCR方法得到的BnRPL基因由3个内含子和4个外显子组成，与拟南芥RPL基因结构相似。氨基酸序列同源性分析表明，BnRPL与拟南芥AtRPL和荒野独行菜（Lepidium campestre）的RPL具有较高的同源性。系统发育进化树显示RPL蛋白在双子叶植物和单子叶植物的分化过程中发生了序列变异。BnRPL基因存在时空表达的组织特异性，在发育的角果表达量最大；成熟后期在角果皮和假隔膜中表达，在种子中不表达。       

花生果种皮特异表达基因AhPSG13的克隆和表达研究

为了克隆在花生果种皮中特异表达的基因,本实验从花生果皮种仁抑制消减文库中筛选出一个277bp的EST序列并进行研究,通过构建花生果皮全长cDNA文库,获得此目的基因G13的全长为1369bp,开放阅读框从第28个碱基始至1122个碱基止,预测分子量为39865.59,等电点5.73。生物信息学分析表明该基因编码的蛋白具有4个跨膜结构域和多个活性位点,可能与细胞内DNA转录有关;该蛋白与多种豆科作物的半胱氨酸蛋白酶具有较高同源性,推测为花生半胱氨酸蛋白酶相关基因;RT-PCR研究该基因表达,结果显示该基因在果种皮中特异表达,于30d果皮中表达量最大。本研究克隆的AhPSG13基因序列已经登陆到GenBank,登录号为FJ475061。     

Mineral composition of some traditional Mexican teas

Teas of plant origin traditionally consumed by the Mountain Pima of Chihuahua, Mexico, were analyzed for mineral nutritional content. Fe, Cu, Zn, Ca, and Mg composition was determined for native teas made from shoots ofTagetes lucida, T. filifolia, Elytraria imbricata, andHolodiscus dumosus, and from root xylem ofCeanothus depressus andPhaseolus ritensis. Native uses of these teas are also described.     

Studies on natural repellents against potato tuber moth (Phthorimaea operculella Zeller) in country stores

Summary The effects of the leaves of five plant species, one from each of the generaAmbrosia, Anemone, Eupatorium, Eucalyptus andLantana, on potato tuber moth were investigated under indigenous storage conditions at the Central Potato Research Station, Shillong (1800 m above sea level). Their action was compared with that of a biological insecticide (spores ofBacillus thuringiensis), a chemical insecticide (carbaryl), and an untreated control. The data collected after six months storage on tuber damage, sprout damage and the rotting indicated that the leaves ofLantana aculeata provided most protection to the tubers, reducing damage from over 70% in the check to below 5%, and sprout damage from over 45% to below 3%. Next best wasEucalyptus globulus followed byB. thuringiensis. They may be used on tubers stored for table use or for seed as they had no adverse effect on germination or on the yield of a subsequent crop.     

Comparative Assessment of the Efficacy of Various Nitrogen Fertilizers

Summary

The efficiency of N fertilizers is usually poor; often less than 50% of the applied N is taken up by the crop. This review focuses on various N fertilizers with respect to the significance of different N loss pathways, namely (i) ammonia volatilization, (ii) dinitrogen and nitrogen oxide emissions, and (iii) nitrate leaching. Further, the significance of biological N immobilization, ammonium fixation and, finally, the impact of nitrate vs. ammonium uptake on crop yield are also discussed. The reviewed literature shows that N fertilizers may differ markedly in their susceptibility to losses. There is, however, considerable scope to improve N efficiency of each N source by proper N management practices.     

Redox Status and Neuro Inflammation Indexes in Cerebellum and Motor Cortex of Wistar Rats Supplemented with Natural Sources of Omega-3 Fatty Acids and Astaxanthin: Fish Oil,Krill Oil,and Algal Biomass

Health authorities worldwide have consistently recommended the regular consumption of marine fishes and seafood to preserve memory, sustain cognitive functions, and prevent neurodegenerative processes in humans. Shrimp, crabs, lobster, and salmon are of particular interest in the human diet due to their substantial provision of omega-3 fatty acids (n-3/PUFAs) and the antioxidant carotenoid astaxanthin (ASTA). However, the optimal ratio between these nutraceuticals in natural sources is apparently the key factor for maximum protection against most neuro-motor disorders. Therefore, we aimed here to investigate the effects of a long-term supplementation with (n-3)/PUFAs-rich fish oil, ASTA-rich algal biomass, the combination of them, or krill oil (a natural combination of both nutrients) on baseline redox balance and neuro-inflammation indexes in cerebellum and motor cortex of Wistar rats. Significant changes in redox metabolism were only observed upon ASTA supplementation, which reinforce its antioxidant properties with a putative mitochondrial-centered action in rat brain. Krill oil imposed mild astrocyte activation in motor cortex of Wistar rats, although no redox or inflammatory index was concomitantly altered. In summary, there is no experimental evidence that krill oil, fish oil, oralgal biomass (minor variation), drastically change the baseline oxidative conditions or the neuro-inflammatory scenario in neuromotor-associated rat brain regions.     

Potential of dry fractionation of wheat bran for the development of food ingredients,part II: Electrostatic separation of particles

Wheat bran is a composite material made of several layers, such as pericarp, testa and aleurone. It could be fractionated into purified fractions, which might either be used as food ingredients, or serve as a starting material for extraction of bioactive compounds. The aim of this work was to evaluate the potential of using electrostatic separation as a way to obtain purified fractions from wheat bran. Ultrafine-ground bran obtained either by cryogenic grinding or by grinding at ambient temperature was used as starting material. The ultrafine bran was then charged by tribo-electrification and introduced in a chamber containing two high voltage electrodes, where bran particles were separated depending on their acquired charge, allowing positively and negatively charged fractions to be collected separately. The particle size distribution, microstructure and biochemical composition of the obtained fractions were studied. The charge of the particles was influenced by their biochemical composition: particles rich in highly branched and cross-linked arabinoxylans (pericarp) were separated from particles rich in β-glucan, ferulic acid and para-coumaric acid (aleurone cell walls). The testa and the intracellular compounds from aleurone were not highly charged, neither positively nor negatively. The most positively charged fraction represented 34% of the initial bran, and contained 62% of the ferulic acid present in the initial bran. The yield of the separation process was good (5.4% loss), and could be further increased.     

In vitro antimicrobial activity of different accessions ofSolanum commersonii Dun. from Uruguay

Summary The in vitro antimicrobial activity of extracts from accessions ofSolanum commersonii Dun. collected in the south of Uruguay was investigated against five microorganisms including the pathogenRalstonia solanacearum. A total of 30 extracts corresponding to organic and aqueous extracts were studied. Interestingly, most of the positive results for growth inhibition were againstR. solanacearum. The extracts were also analyzed for the presence of glycoalkaloids and lectins. Six of the ten aqueous extracts showed lectin presence and a wide variation in the type and amounts of glycoalkaloids, was found. Results indicate that there is no clear relationship between the antimicrobial activity against the five microorganisms screened and the presence or amounts of lectins and glycoalkaloids, traditionally regarded as possible antimicrobial metabolites in theSolanum genus, which indicates the presence of as yet unidentified antimicrobial compounds.     

Pharmacokinetics of Marine-Derived Drugs

Marine organisms represent an excellent source of innovative compounds that have the potential for the development of new drugs. The pharmacokinetics of marine drugs has attracted increasing interest in recent decades due to its effective and potential contribution to the selection of rational dosage recommendations and the optimal use of the therapeutic arsenal. In general, pharmacokinetics studies how drugs change after administration via the processes of absorption, distribution, metabolism, and excretion (ADME). This review provides a summary of the pharmacokinetics studies of marine-derived active compounds, with a particular focus on their ADME. The pharmacokinetics of compounds derived from algae, crustaceans, sea cucumber, fungus, sea urchins, sponges, mollusks, tunicate, and bryozoan is discussed, and the pharmacokinetics data in human experiments are analyzed. In-depth characterization using pharmacokinetics is useful for obtaining information for understanding the molecular basis of pharmacological activity, for correct doses and treatment schemes selection, and for more effective drug application. Thus, an increase in pharmacokinetic research on marine-derived compounds is expected in the near future.     

Novel food and non-food uses for sorghum and millets

Sorghum and millets have considerable potential in foods and beverages. As they are gluten-free they are suitable for coeliacs. Sorghum is also a potentially important source of nutraceuticals such antioxidant phenolics and cholesterol-lowering waxes. Cakes, cookies, pasta, a parboiled rice-like product and snack foods have been successfully produced from sorghum and, in some cases, millets. Wheat-free sorghum or millet bread remains the main challenge. Additives such as native and pre-gelatinised starches, hydrocolloids, fat, egg and rye pentosans improve bread quality. However, specific volumes are lower than those for wheat bread or gluten-free breads based on pure starches, and in many cases, breads tend to stale faster. Lager and stout beers with sorghum are brewed commercially. Sorghum's high-starch gelatinisation temperature and low beta-amylase activity remain problems with regard to complete substitution of barley malt with sorghum malt . The role of the sorghum endosperm matrix protein and cell wall components in limiting extract is a research focus. Brewing with millets is still at an experimental stage. Sorghum could be important for bioethanol and other bio-industrial products. Bioethanol research has focused on improving the economics of the process through cultivar selection, method development for low-quality grain and pre-processing to recover valuable by-products. Potential by-products such as the kafirin prolamin proteins and the pericarp wax have potential as bioplastic films and coatings for foods, primarily due to their hydrophobicity.