大豆转座子的研究现状及应用前景

大豆基因组中约有5万基因,大部分基因的功能处于未知状态。转座子插入突变体库的应用可以为解码大豆基因组中未知基因的功能提供研究平台,同时,也可以为创建有育种应用价值的大豆突变体种质资源库提供基础。大豆基因组中50%~60%的组分为转座子,其中具有活性的且结构完整的转座子只有Tgm9。大豆转座子突变体库目前主要还是利用外源转座子来建立,但是开发并改进内源转座子,增加其跳跃能力及稳定性,可以成为构建突变体库的一个新方向。本文主要介绍了大豆基因组中发现的各种转座子及其特点,并且重点讨论了外源转座子和大豆内源转座子在大豆突变体库创建中的应用现状及应用前景。     

Ac/Ds转座子系统及其在玉米突变体库构建中的应用

Ac/Ds转座子系统是研究玉米功能基因组学的重要工具。Ac/Ds转座子系统以其在基因组中的低拷贝、倾向插入基因外显子、可以在同一基因不同位点造成突变等独特优势,在构建玉米突变体库和进行基因功能鉴定方面具有重要应用前景。综述了Ac/Ds转座子系统的插入特点、转座机制、在玉米突变体库构建中的应用以及我国玉米突变体库的现状,并对我国玉米Ac/Ds突变体库的研究提出展望。     

诱变育种技术在大豆育种与基因挖掘中的应用现状及展望

植物诱变育种能够创造许多优异变异资源,诱变获得的突变体可以作为种质材料,为大豆新品种培育提供丰富的资源;构建的突变体库也有助于大豆功能基因组研究的开展,可为特定性状的研究提供遗传材料。本文介绍诱变育种技术在大豆生长特性、品质和抗性改良育种中的应用,分析利用诱变突变体库进行大豆生长特性、品质性状改良和抗性相关基因挖掘的研究进展,介绍诱变技术与其他生物技术相结合发掘目的基因的应用现状,并展望今后大豆诱变育种技术在基因挖掘方面的应用前景,以期能更好地运用诱变育种技术推动大豆遗传研究和品种选育。     

转座子标签法及其在大豆功能基因组研究中的应用前景

转座子标签法是克隆未知基因的有效方法之一.本文介绍了转座子标签法的基本原理、步骤、方法及其在植物基因克隆中的应用情况,并对该方法在大豆突变体库构建和功能基因组研究中的应用前景进行了讨论.     

大豆EMS突变体库构建及其M_1代重要农艺性状评价

构建突变体库是进行遗传学与分子生物学研究的重要前提条件之一。当前大豆突变体数量的不足,严重制约了大豆功能基因组学研究的深入开展。以南方大面积推广的大豆品种-南豆12为材料,通过EMS诱变构建了大豆EMS突变体库。统计发现,M1种子田间出苗率为34.85%,表明EMS强烈抑制大豆种子的萌发和出苗。对随机选取的913株M1植株进行了株高、单株荚数等重要农艺性状的调查。结果表明:经EMS处理后,M1植株在株高、单株荚数、节间数等方面与对照存在显著差异、表型变异丰富,其中18.07%发生矮化,11.06%表现出多荚,另有3株不育株;群体间的节间个数极差达到18。这些数据表明已成功构建了大豆EMS突变体库。在进一步的研究中,将对单个突变体进行分离、分类,并克隆相应的重要功能基因,为深入研究大豆功能基因组学以及筛选适合玉米-大豆带状复合种植模式的高光效、抗倒伏大豆品种奠定基础。     

EMS诱发大豆“南农94-16”突变体库的扩建及部分突变体的SSR分析

甲基磺酸乙酯(EMS)是一种稳定、高效的诱发点突变的诱变剂.采用EMS处理大豆"南农94-16"的种子,在初步构建大豆突变体库的基础上进一步扩建了大豆突变体库,获得95份茎、叶、花、种子等性状变异的突变材料.采用200对SSR标记分别对其中高蛋白、叶黄化等9个不同性状的突变体进行了鉴定.结果表明:与对照相比突变体发生了多位点的变异.构建的突变体库不但可以作为新的种质资源,而且可促进大豆功能基因组研究的开展.     

甘蓝型油菜花序长发夹ｌｈＲＮＡｉ文库的构建

异源四倍体油菜含有大量多拷贝基因及冗余基因,难以通过T-DNA插入、物理及化学诱变等常规突变体库创制方法发掘功能基因。为开发适应于油菜的简单高效的突变体库创制方法,本研究通过滚环复制方法构建了甘蓝型油菜的花序lhRNAi文库,并对这种新型的干扰文库进行质量评估;然后将该花序lhRNAi文库对甘蓝型油菜进行遗传转化,获得763株T0植株,从中鉴定分离出74株具有可见表型变异的花发育相关突变体,包括花瓣减少、形状异常,柱头卷曲,雄蕊退化萎缩,雄性不育（不能产生花粉）、死蕾或花蕾闭合等。说明通过滚环复制介导的lihRNAi文库的构建方法可以成功应用到油菜中,这将为油菜花序发育相关基因功能的研究提供重要的研究平台。     

大豆“冀黄13”突变体筛选及突变体库的建立

利用甲基磺酸乙酯(ethyl methane sulfonate,EMS)、叠氮化钠(sodium azide,NaN3)和N离子束分别诱变处理大豆品种"冀黄13"的种子。经M2选择,M3、M4代验证,共筛选出茎器官突变体15份,叶器官突变体19份,花器官突变体31份,种子器官突变体3份以及成熟期突变体18份,其中有9份突变体属于复合性状的突变,最终获得77份稳定遗传的突变体。通过F2群体的表型分析,确定1份雄性不育突变体和1份黑种皮色突变体的表型均由单隐性基因控制。本研究所获得的突变体和所构建突变体库将为大豆功能基因组学研究和遗传改良奠定基础。     

木薯细菌性枯萎病菌转座子库的构建及其质量评价

为了研究木薯细菌性枯萎病菌的致病分子机理,本研究开展了病原菌的转化子库构建工作.采用电击法,将EZ::TN转座体转化野生型菌株.获得了总数为20 382个转化子并进行了质量评价,采用剪叶法从9 872个转化子中筛选出94个致病性变异的转化子.采用Tail-PCR获得了致病力丧失突变体Xtn62-36的侧翼序列,分析结果表明可能是外源片段插入预测基因AspCxam的编码区而造成致病力丧失.病原菌转化子库的构建及Tail-PCR技术的应用为进一步开展致病相关基因功能的研究提供了基础.     

病毒诱导的基因沉默(VIGS)技术及其在大豆基因功能研究和育种中的应用潜力

病毒诱导的基因沉默(virus-induced gene silencing,VIGS)是植物体抵抗病毒侵染的一种自然防御机制,近年来已发展成为一种基因功能研究的有效手段。与突变体筛选、转基因、基因敲除等技术相比,VIGS技术因具有周期短、成本低、可迅速观察表型等优点而被广泛应用于基因的功能鉴定。文章对VIGS技术的分子机理、病毒载体系统、稳定遗传VIGS植株的获得策略及其在大豆中的应用等内容进行了综述并对今后的发展趋势进行了讨论。     

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.     

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.     

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.     

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.     

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.