海莲耐盐基因AOC克隆及转录融合表达载体的构建

利用RT-PCR的方法从红树林海莲中克隆丙二烯环化氧化酶基因(AOC),并进行序列测定和BLAST程序进行同源序列分析。从RT-PCR得到的产物,其核苷酸序列与GeneBank中海莲丙二烯环化氧化酶基因AO(CBAB21610)的核苷酸序列完全相同。AOC基因通过中间载体pBS-AOC,pJIT60-AOC,最后克隆到植物表达载体pGreenII0229上,得到双35S的pGreenII0229-AOC植物表达载体。AOC基因和ipt基因通过中间载体pBS-AOC,pBS-ipt,pBS-AOC-ipt,pJIT60-AOC-ipt;最后将AOC和ipt基因以转录融合的方式克隆到植物表达载体pGreenII0229上,得到具有双35S启动子的AOC,ipt双基因转录融合的植物表达载体pGreenII0229-AOC-ipt。     

GmAOC3基因转化载体构建及转化大豆的初步研究

选用内切酶SacⅠ和MluⅠ切割目标基因和载体,构建了携带bar基因的重组载体p BA002-Gm AOC3,选用2个大豆品种(Jack和南农88-1)和2种外植体(子叶节和整个子叶节),研究大豆品种和外植体对根癌农杆菌介导的子叶节转化Gm AOC3基因的影响。结果表明:外植体为整个子叶节的大豆品种Jack的出芽率和转化率最高,分别为79.5%和2.27%。经Quick Stix PAT/bar基因试纸条检测、目标基因的分子检测和草丁膦抗性鉴定,共得到转Gm AOC3基因的T0代阳性株系3株,T1代阳性转基因大豆6株,其中5株以Jack品种为受体的转基因大豆Gm AOC3基因的表达量均显著高于非转基因植株,荧光定量拷贝数检测结果显示,4株转基因单株为单拷贝,2株为双拷贝。     

甘蔗丙二烯氧化物环化酶基因（ScAOC）的克隆与表达分析

丙二烯氧化物环化酶(allene oxide cyclase,AOC)是茉莉酸合成途径的一个关键酶,在植物防御反应中起着重要的作用。本研究基于甘蔗转录组数据库克隆到一个AOC基因,命名为ScAOC,并对其进行生物学分析、组织特异性分析和逆境胁迫下的表达分析,以初步了解该基因的功能。甘蔗ScAOC基因的开放阅读框(ORF)为744 bp,编码244个氨基酸,其蛋白分子质量是26.37 kDa。生物信息学分析结果显示,ScAOC蛋白属于不稳定碱性亲水蛋白,含有1个Allene_ox_cyc Superfamily的PLN02343 domain的保守结构域;构建系统进化树,结果显示ScAOC蛋白与高粱的蛋白具有较高的同源性。荧光定量PCR结果显示,ScAOC基因在甘蔗的根、茎、叶中均有表达,表达量从高到低依次是叶、根、茎。在PEG、NaCl及外源MeJA胁迫下,ScAOC基因相对表达量都是先上升后下降,而ABA对ScAOC的表达有一定的抑制作用。在病原菌和粘虫取食的胁迫下,ScAOC基因相对表达量显著上升。结果显示ScAOC可能参与甘蔗对生物或非生物...     

桃蚜取食抗、感辣椒品种的茉莉酸途径基因表达差异分析

根据蚜口密度及取食时间效应,明确桃蚜取食前后辣椒茉莉酸（JA）合成及信号途径基因表达差异,初步阐明JA途径介导的辣椒抗蚜性分子机理。本研究以抗蚜辣椒品种猪大肠（ZDC）和感蚜辣椒品种大羊角椒（DYJJ）为材料,采用实时定量PCR技术分析与植物防御相关的JA合成途径基因（LOX2、AOS、AOC和OPR3）与信号途径基因（COI1和JAR1）在桃蚜不同蚜口密度（20、30、40、50、60蚜/叶）及取食不同时间（6、12、24、48、72 h）后的表达量变化情况。结果表明：在不同蚜口密度和取食为害时间下,JA合成途径的4个基因在ZDC中表达量总体上显著高于DYJJ,而信号途径2个基因表达量总体上显著低于DYJJ。在桃蚜较低为害水平状态下,抗、感辣椒品种之间表达量差异显著,且在部分相同蚜口密度下部分基因表达量峰值和低谷出现时间基本一致。较高水平的为害情况下,抗、感辣椒品种之间差异显著性下降。以上结果表明,JA合成及信号途径AOS、AOC及JAR1基因可能具有作为鉴定评价辣椒抗蚜性水平的分子指标的潜力,可为将来辣椒抗蚜种质资源评价与创新利用提供理论依据。     

生物与非生物胁迫下水稻CIPK基因的鉴定分析

类钙调磷酸酶B亚基蛋白(calcineurin B-like proteins,CBL)是植物细胞中重要的Ca2+传感器,必须与蛋白激酶CIPK(CBL-interacting protein kinases)特异性互作才能发挥生物学功能.根据水稻OsCIPK(Oryza sativa CIPK)基因家族的预测序列,从粳稻日本晴(Nipponbare)中克隆了15个OsCIPK基因.基因结构分析表明15个OsCIPK可归为多内含子和少内含子两类,OsCIPK3和OsCIPK24的mRNA存在可变剪接.系统进化树分析显示水稻CIPK家族与拟南芥、杨树来源于同一个祖先.这15个OsCIPK在不同程度上受生物胁迫(白叶枯病)和非生物胁迫(Hg2+、高盐、冷和ABA)的诱导表达;其中,5个基因(OsCIPK1、OsCIPK2、OsCIPK10、OsCIPK11和OsCIPK12)在接种白叶枯病菌后表达显著上调;而4个基因(OsCIPK2、OsCIPK10、OsCIPK11和OsCIPK14)的表达受所有胁迫处理诱导,表明这些基因可能参与多种逆境信号的转导.揭示了CIPK可能在植物的生物和非生物胁迫应答中都具有重要的作用.     

河南省花生品种(系)基于SSR分子标记的遗传多样性研究

花生是我国重要的经济作物和油料作物。对河南省不同类型花生品种的遗传多样性进行分析、评价具有重要意义。本研究利用200对多态性较好的SSR分子标记对河南省近年来正在推广及新选育的60个花生品种(系)进行遗传多样性研究,品种(系)间相似性分析结果表明:平均遗传相似系数为0.3,在实验分析的1770个关系组合中,仅有616个遗传相似系数大于0.5,说明这些品种大多具有较低的遗传相似性。聚类结果表明,在距离320处,60份花生品种(系)被分成了5大类群,第一类群含有32个品种,第二、第三类群分别含有8个品种,第四类含有3个品种,第五类群含有9个品种。这五大类群中,均含有来源于郑州的品种。研究结果也表明,开19-2、商花10号和开农012具有独特的遗传背景,是重要的花生资源。本研究对60个河南省花生品种(系)的遗传多样性的分析,可以为花生新品种选育提供重要的理论参考。     

2009-2010年河南、河北、山东三省小麦区试品种（系）的亲缘系数分析

为了解2009-2010年度河南、河北及山东参加区试品种(系)小麦的遗传背景丰富度,追溯了这三个省份供试的85个小麦品种(系)信息,采用亲缘系数(coefficient of parentage,COP)和聚类分析的方法对其进行分析.结果表明,在供试的85个品种(系)的3 570对组合中,所有组合的COP值变异范围为0.000～0.750,亲缘系数总和为77.244,平均值为0.021.聚类分析将这85个品种(系)聚为8类.三个省份中河南参试的品种(系)遗传多样性相对较低,主要因为这些品种(系)是利用豫麦2号及其衍生品种(系)培育而成的;河北、山东参加区试的小麦品种(系)遗传多样性相对较高,主要由于这些地区广泛利用了当地的种质资源,其中山东省小麦品种培育主要选用了20世纪50年代以来生产上比较重要的多个品种,包括鲁麦14、鲁麦23、泰山21号及济南13号的衍生品种莱州95021等.在以后的育种工作中应加强对重要小麦品种(系)的应用及引进新的种质资源.     

盐胁迫对马铃薯出苗及苗期生物学性状的影响

马铃薯作为重要的粮、菜兼用型作物,深受南疆少数民族的喜爱.马铃薯属盐敏感型作物,为研究其耐盐机制,选育耐盐品种,试验采用盆栽方法,研究了4个马铃薯品种在土壤含盐量(NaCl)分别为0(对照)、0.3％(处理1)、0.6％(处理2)条件下的出苗天数、出苗率、苗期株高、SPAD值、过氧化氢酶(CAT)活性、过氧化物酶(PO...     

西南地区小麦种质资源白粉病抗性的全基因组关联分析

为了有效利用我国西南地区小麦抗白粉病种质资源,于2012―2013年在贵州贵阳、贵州赫章和四川绵阳以及2013―2014年在贵州贵阳对主要来自西南地区的120个小麦品种(系)进行了白粉病抗性鉴定,并利用小麦90K芯片进行全基因组关联分析。抗病性鉴定结果显示,36个品种(系)在四个环境下均表现出抗性,其中大部分品种(系)来自贵州省。全基因组关联分析发现,16个SNP位点与白粉病抗性显著相关,分别位于1A、1D、2A、3A、4B、5B、6A、6D和7B染色体上,其中位于6AS染色体重要区段上的3个SNP位点(Jagger_c4823_169、Tdurum_contig55201_928和Tdurum_contig12215.89)的遗传距离小于1cM,在四个环境中均与白粉病抗性显著相关。相关分析表明,这3个SNP位点与 Pm21分子标记(SCAR1265)紧密连锁。     

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.     

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.     

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.     

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.     

Extreme resistance to infection by potato virus X in genotypes of wild tuber-bearingSolanum species

Summary Clones derived from thirty-one different accessions (nineteen of Argentine origin) belonging to eightSolanum species were screened for resistance to infection by potato virus X strain cp (PVX cp) by mechanical inoculation of plantlets that had been micropropagated in vitro. Estimates of PVX multiplication obtained by enzyme linked immunosorbent assay and slot blot nucleic acid hybridization allowed the identification of resistant clones derived from five accessions belonging toS. commersonii S. oplocense, S. sparsipilum andS. tuberosum andigena. Resistant genotypes supported PVX concentrations 5 to 15 times smaller than did the susceptible control cultivar Spunta. Graft inoculation test confirmed the presence of extreme resistance similar to that conferred by the ‘immunity’ gene X¹ (also called RX_act).     

Cyanobacteria Secondary Metabolites as Biotechnological Ingredients in Natural Anti-Aging Cosmetics: Potential to Overcome Hyperpigmentation,Loss of Skin Density and UV Radiation-Deleterious Effects

The loss of density and elasticity, the appearance of wrinkles and hyperpigmentation are among the first noticeable signs of skin aging. Beyond UV radiation and oxidative stress, matrix metalloproteinases (MMPs) assume a preponderant role in the process, since their deregulation results in the degradation of most extracellular matrix components. In this survey, four cyanobacteria strains were explored for their capacity to produce secondary metabolites with biotechnological potential for use in anti-aging formulations. Leptolyngbya boryana LEGE 15486 and Cephalothrix lacustris LEGE 15493 from freshwater ecosystems, and Leptolyngbya cf. ectocarpi LEGE 11479 and Nodosilinea nodulosa LEGE 06104 from marine habitats were sequentially extracted with acetone and water, and extracts were analyzed for their toxicity in cell lines with key roles in the skin context (HaCAT, 3T3L1, and hCMEC). The non-toxic extracts were chemically characterized in terms of proteins, carotenoids, phenols, and chlorophyll a, and their anti-aging potential was explored through their ability to scavenge the physiological free radical superoxide anion radical (O₂^•−), to reduce the activity of the MMPs elastase and hyaluronidase, to inhibit tyrosinase and thus avoid melanin production, and to block UV-B radiation (sun protection factor, SPF). Leptolyngbya species stood out for anti-aging purposes: L. boryana LEGE 15486 presented a remarkable SPF of 19 (at 200 µg/mL), being among the best species regarding O₂^•− scavenging, (IC₅₀ = 99.50 µg/mL) and also being able to inhibit tyrosinase (IC₂₅ = 784 µg/mL), proving to be promising against UV-induced skin-aging; L. ectocarpi LEGE 11479 was more efficient in inhibiting MMPs (hyaluronidase, IC₅₀ = 863 µg/mL; elastase, IC₅₀ = 391 µg/mL), thus being the choice to retard dermal density loss. Principal component analysis (PCA) of the data allowed the grouping of extracts into three groups, according to their chemical composition; the correlation of carotenoids and chlorophyll a with MMPs activity (p < 0.01), O₂^•− scavenging with phenolic compounds (p < 0.01), and phycocyanin and allophycocyanin with SPF, pointing to these compounds in particular as responsible for UV-B blockage. This original survey explores, for the first time, the biotechnological potential of these cyanobacteria strains in the field of skin aging, demonstrating the promising, innovative, and multifactorial nature of these microorganisms.     

Marine Pharmacology in 2009–2011: Marine Compounds with Antibacterial,Antidiabetic, Antifungal,Anti-Inflammatory,Antiprotozoal, Antituberculosis,and Antiviral Activities; Affecting the Immune and Nervous Systems,and other Miscellaneous Mechanisms of Action ?

The peer-reviewed marine pharmacology literature from 2009 to 2011 is presented in this review, following the format used in the 1998–2008 reviews of this series. The pharmacology of structurally-characterized compounds isolated from marine animals, algae, fungi and bacteria is discussed in a comprehensive manner. Antibacterial, antifungal, antiprotozoal, antituberculosis, and antiviral pharmacological activities were reported for 102 marine natural products. Additionally, 60 marine compounds were observed to affect the immune and nervous system as well as possess antidiabetic and anti-inflammatory effects. Finally, 68 marine metabolites were shown to interact with a variety of receptors and molecular targets, and thus will probably contribute to multiple pharmacological classes upon further mechanism of action studies. Marine pharmacology during 2009–2011 remained a global enterprise, with researchers from 35 countries, and the United States, contributing to the preclinical pharmacology of 262 marine compounds which are part of the preclinical pharmaceutical pipeline. Continued pharmacological research with marine natural products will contribute to enhance the marine pharmaceutical clinical pipeline, which in 2013 consisted of 17 marine natural products, analogs or derivatives targeting a limited number of disease categories.     

Recent Modifications of Chitosan for Adsorption Applications: A Critical and Systematic Review

Chitosan is considered to be one of the most promising and applicable materials in adsorption applications. The existence of amino and hydroxyl groups in its molecules contributes to many possible adsorption interactions between chitosan and pollutants (dyes, metals, ions, phenols, pharmaceuticals/drugs, pesticides, herbicides, etc.). These functional groups can help in establishing positions for modification. Based on the learning from previously published works in literature, researchers have achieved a modification of chitosan with a number of different functional groups. This work summarizes the published works of the last three years (2012–2014) regarding the modification reactions of chitosans (grafting, cross-linking, etc.) and their application to adsorption of different environmental pollutants (in liquid-phase).