一种适用于多糖多酚植物的高质量RNA快速提取方法

植物总RNA的提取是进行下游分子生物学实验的关键基础,高质量的RNA是下游实验成功的保障。许多植物由于其体内含有酚、多糖以及其他的一些次生代谢物,要获得这些植物高质量的总RNA比较困难。本文在多次实验的基础上,提出了改良LiCl沉淀法,该方法步骤少、效率高,使用此方法得到的棉花幼苗总RNA经检测表明其质量比使用冷酚法提取的RNA要高得多,且完全满足后续实验的要求。使用此方法成功提取了香蕉叶片、根、木薯叶片的RNA,经检测表明,所得到的RNA完整性好,完全可以满足后续实验的要求,说明改良LiCl沉淀法是一种适用于多酚多糖植物的快速RNA提取方法。     

影响植物组织总RNA质量的因素

随着大量生物领域科研人员进入植物RNA研究领域，开展RNA研究，使得以RNA为终产物的基因组研究取得突破性进展，新的RNA基因不断增加，RNA的新功能不断被发现。但由于RNA的不稳定性，使得RNA操作远比DNA操作困难，且尚有大量未知的RNA及其功能等待人们去探索、研究．而这些研究又都基于高质量RNA的提取。因此，根据笔者自身实验经验，及在查阅大量相关文献的基础上。针对植物总RNA提取的一些关键影响因素、消除对策、RNA检测方法及RNA保存方法上进行了系统的阐述，旨在为高质量植物组织总RNA的制备提供理论参考，为RNA分子生物学实验后续研究提供理论支撑。     

香蕉果实RNA提取方法的改进和比较

通过对几种不同的香蕉果实RNA提取方法的改进和比较,发现CTAB法是最有效、最简单和最经济的方法之一.该方法提取出来的RNA可以用来做cDNA合成和建立文库等分子生物学操作.     

RNA干扰技术及其在茶树中的应用前景

RNA干扰（RNA interference,RNAi）是指由双键RNA（double-stranded RNA,dsRNA）启动的序列特异的转录后基因沉默现象,广泛存在于真菌、植物和动物中。本文综述了RNA干扰技术的发展简史、分子机制、作用特点以及在植物中的应用,并就RNAi技术在茶树中的应用前景做了简要分析。     

甘蔗原生质体RNA提取条件和方法的优化

为了获得高产量和高纯度的甘蔗原生质体RNA,本研究以新台糖22号（ROC22）和桂糖28号（GT28）甘蔗原生质体为材料,探究了渗透压和原生质体活力对甘蔗原生质体RNA提取效果的影响,并比较了Trizol法、改良Trizol法、改良CTAB法、试剂盒法4种RNA提取方法。结果表明：原生质体渗透压与RNase活性呈显著正相关,原生质体活力与RNase活性呈极显著负相关;RNase活性越高,RNA提取效果越差;使用0.5 mol/L的甘露醇作为渗透压调节剂可获取活力最高的甘蔗原生质体;原生质体活力为70%和90%时所提的RNA完整性好且纯度高,符合后续分子实验的需求,原生质体活力为90%时,RNA产量最高。因此,需要提取甘蔗原生质RNA时,至少需要保证原生质体的活力在70%以上,且原生质体活力越高,RNA的产量越高;采用改良Trizol法,可以大量制备能满足后续分子实验的完整性好且纯度高的RNA,利用该条件和方法,可以确保获得大量高质量的甘蔗原生质体RNA。本研究结果为甘蔗体细胞融合育种过程中的分子生物学研究提供了方法和技术支撑。     

冻融法纯化水稻胚乳RNA及稻米品质重要相关基因表达谱研究

 提出了一种从水稻胚乳组织中快速分离总RNA的方法，可以简便地去除RNA中存在的淀粉等多糖类杂质，所获的胚乳总RNA其质量可满足Northern印迹、微阵列制备等分子生物学实验要求。将从966个水稻品种的胚乳中分离所得的RNA制备RNA阵列，用于稻米品质重要基因Wx基因的表达谱分析，结果证实：水稻Wx基因表达丰度总体上与水稻胚乳的直链淀粉含量呈正相关；应用于Northern印迹实验，表明在水稻胚乳组织中，淀粉分支酶Rbe1具有两个长度不同的同源基因，其中一个分子量大的为胚乳特异表达，分子量小的则在水稻各组织中共表达。     

应用重组PCR技术构建大豆脂肪氧化酶基因RNA干扰表达载体

RNA干扰是近年来生物技术领域新兴的研究热点之一。在作物的研究中,RNA干扰被认为在作物品质改良中具有重要的应用价值,并已在水稻、小麦、玉米等作物中得到成功应用,但在大豆的研究中尚鲜有报道。在研究植物RNA干扰的实验中,构建相应的RNA干扰表达载体是常用手段,但传统构建方法较费时费力。重组PCR是一种利用重叠延伸的方法进行高效和快速的体外基因片段拼接的PCR技术。研究将重组PCR技术应用于构建大豆脂肪氧化酶基因ihpRNA干扰表达载体,一方面探索一条更简便、快捷、适应范围更广泛的大豆基因RNA干扰表达载体构建方法,另一方面所构建的载体为进一步研究RNA干扰在大豆脂肪氧化酶改良中的应用奠定了基础。结果显示应用重组PCR技术构建大豆脂肪氧化酶基因RNA干扰表达载体是可行的,相比与常规方法此方法更简便、快捷、效率更高。     

茶树中提纯总RNA的研究

在克氏一步法分离总RNA的基础上 ,根据茶树叶片生化成分特点 ,做了部分修改 ,并研究了不同浓度的L -半胱氨酸和可溶性聚乙烯吡咯烷酮 (PVP)、不溶性PVP对提取的RNA纯度、回收率和完整性的影响。分光光度计检测和变性琼脂糖凝胶电泳结果证明 :使用本方法并加入 5- 10mmol LL -半胱氨酸或 4 % - 6 %可溶性PVP能提高RNA纯度和 (或 )回收率 ,研磨时加入5% - 10 %不溶性PVP对提高RNA完整性有较好的效果     

2种微藻总RNA提取方法的比较

以微茫藻和莱茵衣藻为材料，比较Trizol试剂法和SDS-LiCl沉淀法提取2种微藻总RNA的效果，以期筛选出适合微藻RNA的提取方法。结果表明，SDS-LiCl沉淀法能从微茫藻中提取高质量的RNA，而Trizol法和SDS-LiCl沉淀法均能从莱茵衣藻中获得高质量的RNA，但Trizol法更为简单有效。RT-PCR结果表明，SDS-LiCl沉淀法提取的微茫藻总RNA和Trizol法提取的莱茵衣藻总RNA都能直接用于分子克隆和基因表达分析等分子生物学实验。     

橡胶树胶乳RNA简易提取和保存方法的比较探索

针对巴西橡胶树胶乳RNA的提取方法存在操作复杂、步骤多、时间长、保存条件要求苛刻等问题以及橡胶树胶乳远距离、长时间采样的诸多不便等问题，介绍一种具有操作简单、实验时间短、步骤少等优点的简易巴西橡胶树胶乳RNA提取方法和一种适于长时间保存胶乳的方法。该方法仅仅以DEPC水为提取液，提取的RNA浓度可以达到为0.722 μg/mL，纯度OD260/OD280在2.0～2.2之间，总RNA提取量为36.1 μg/mL，可以满足后续分子生物学实验RT-PCR和RACE的要求；对在冰上保存1～10 d的胶乳提取RNA，浓度在0.502～0.722 μg/mL之间，OD260/OD280在1.8～2.4之间，总RNA的提取量在20.8～35.44 μg/mL之间。     

In vitro reproduction in the annual pasture legumes subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.)

Pasture legumes are important components of both mixed farming rotations and permanent pastures in temperate climates. Breeding of two widely sown pasture legumes, subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.), is constrained by the long generation cycle, typically enabling only one generation per year. We hypothesized manipulation of culture medium and conditions would enable the development of a laboratory‐based protocol for in vitro reproduction in subterranean clover and French serradella. In vitro flowering and viable seed set was induced from both species. For subterranean clover, the most effective treatment was culturing on modified MS medium with 1 μm kinetin and 0·1 m sucrose under a 100 μmol m^?2 s^?1 light intensity and continuous photoperiod. For French serradella, culture on a hormone‐free B5 medium with 5 mm NH₄Cl and 0·1 m sucrose under a 100 μmol m^?2 s^?1 light intensity and 20 h photoperiod was optimum. It is expected this technique will have application in accelerating generation turnover within breeding programs, for the study of factors influencing flowering in pasture legumes, and for the propagation of valuable yet enfeebled plants such as embryo‐rescued hybrids.     

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.     

Estimation of lucerne yield stability for enabling effective cultivar selection under rainfed conditions

Evaluation of crop and forage yield stability is of increasing relevance in the context of current and recent environmental changes but, in contrast to other field crops, there are no published systematic analyses among forage crops in Europe. A study of stability performance was conducted with 13 Czech cultivars of lucerne at four locations over a 2‐year period with the following aims: (a) to evaluate yield stability of varieties across different environments and (b) to calculate measurable benefits of variety selection in relation to the specific environment. The cultivar Vlasta was identified as the highest yielding cultivar (annual yield 16.0 t DM/ha), whereas the lowest yielding cv. Magda, Tereza and Oslava averaged around 14.9 t DM/ha. Effect of genotype × environment interaction (G × E) was two times higher than for genotype alone. Additive main effects and the multiplicative interaction (AMMI) model showed that the highest yielding cultivars may not be stable across environments. This study demonstrated further that significant yield improvement could be detected, even among a relatively homogenous group of domestic cultivars, and this was driven mainly by site productivity: the improvement was +10% in low‐yielding sites, compared with +3% in high‐yielding sites. Results highlight that advanced agronomy should also consider stability parameters such as AMMI stability value or superiority measures for forage crops in response to the challenges associated with climate change.     

La pourriture fongique des tubercules de pomme de terre en atmosphère contrôlée

Résumé Fusarium culmorum et, dans une moindre mesure,F. avenaceum, F. sambucinum, Colletotrichum coccodes etCylindrocarpon sp. ont été isolés de tubercules entreposés en atmosphère contr?lée (AC) et atteints de pourriture grave. Des expériences d'inoculation de tubercules avec 10 espèces ou variétés fongiques et conservation en AC à 8 et 10°C ont montré que: 1) les champignons non pathogènes en atmosphère normale (ex.:Cylindrocarpon sp.) le devenaient en AC, 2) ceux qui étaient faiblement pathogènes en atmosphère normale étaient nettement plus virulents en AC (ex.:F. culmorum), 3) les espèces ou variétés virulentes en conditions d'air normales (Fusarium sulphureum etPhoma exigua var.foveata) ne provoquaient pas de pourriture plus grave en AC et leur activité pouvait même être légérement freinée.

---

Summary The extensive rotting of tubers which occurred during storage trials in controlled atmospheres has led to a mycological and pathological study of the rots. The most frequently isolated species wasFusarium culmorum althoughF. avenaceum, F. Sambucinum, Coletotrichum coccodes and aCylindrocarpon sp. were also present. Tubers of cvs Bintje and Ulla were inoculated with 19 isolates of 10 fungal species or varieties associated with rotting of stored tubers (Table 1). The tubers were incubated at 8 or 10°C in normal or controlled (2% O₂:1% CO₂, 2% O₂:6% CO₂, 2% O₂:12% CO₂, 6% O₂:6% CO₂) atmospheres and the extent of rotting was determined a month later (Tables 2 and 3 and Fig. 2). Species which were weakly virulent in a normal atmosphere (variousFusarium spp. andPhoma exigua var.exigua) were clearly more aggressive in the controlled atmosphere and this was especially evident in the case ofF. culmorum. Species ot pathogenic in a normal atmosphere under experimental conditions (Colletotrichum coccodes, Cylindrocarpon sp.,Phoma eupyrena) caused rotting in the controlled atmosphere. The two species which were virulent in a normal atmosphere (F. sulphureum andP. exigua var.foveata) were also virulent in the controlled atmosphere and in some cases the extent of rotting was even less. As a general rule, rotting caused by weakly virulent fungi was greater in both cultivars in the controlled than in a normal atmosphere.

---

Zusammenfassung Eine starke F?ulnis von in Kontrotrollierter Atmosph?re gelagerten Kartoffelknollen veranlasste eine mykologische und pathologische Untersuchung. Der am h?ufigsten isolierte Pilz warFusarium culmorum. F. avenaceum, F. sambucinum, Colletotrichum coccodes undCylindrocarpon sp. wurden ebenfalls bestimmt. Neunzehn St?mme von 10 Pilzsorten oder-Arten welche bei Lagerf?ulnis vorkommen (Tab. 1) wurden auf Knollen der Sorten Bintje und Ulla inokuliert. Die Lagerung wurde bei 8 oder 10°C under Normalbedingungen oder kontrollierter Atmosph?re durchgeführt (2% O₂:1% CO₂, 2% O₂:6% CO₂, 2% O₂:12% CO₂, 6% O₂:6% CO₂). Der F?ulnisgrad (Abb. 1) wurde nach einem Monat bonitiert. Die Ergebnisse sind in den Tabellen 2 und 3 dargestellt und in der Abb. 2 illustriert. Unter in Normalbedingungen nur schwach virulente Arten (verschiedeneFusarien undPhoma exigua var. exigua) waren deutlich aktiver in kontrollierter Atmosph?re. Dies was besonders der Fall fürF. culmorum. Pilze, die im Versuch unter Normalbedingungen nicht pathogen waren (Colletotrichum coccodes, Cylindrocarpon sp., Phoma eupyrena), haben unter kontrollierter Atmosph?re Sch?den verursacht. Hingegen werden die beiden unter Normalbedingungen sehr pathogenen Arten (F. sulphureum undP., exigua var.foveata), durch kontrollierte Atmosph?re wenig beeinflusst. Es wurde sogar in einigen F?llen eine gewisse Verringerung des F?ulnisbefalls beobachtet. Ganz allgemein haben die getesteten kontrollierten Bedingungen die Entwicklung der Pilzf?ulnis an beiden Kartoffelsorten gef?rdert, speziell durch Pilzarten, welche unter Normal-bedingungen wenig Sch?den verursachen.

     

Marine Pharmacology in 2016–2017: 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 review of the 2016–2017 marine pharmacology literature was prepared in a manner similar as the 10 prior reviews of this series. Preclinical marine pharmacology research during 2016–2017 assessed 313 marine compounds with novel pharmacology reported by a growing number of investigators from 54 countries. The peer-reviewed literature reported antibacterial, antifungal, antiprotozoal, antituberculosis, and antiviral activities for 123 marine natural products, 111 marine compounds with antidiabetic and anti-inflammatory activities as well as affecting the immune and nervous system, while in contrast 79 marine compounds displayed miscellaneous mechanisms of action which upon further investigation may contribute to several pharmacological classes. Therefore, in 2016–2017, the preclinical marine natural product pharmacology pipeline generated both novel pharmacology as well as potentially new lead compounds for the growing clinical marine pharmaceutical pipeline, and thus sustained with its contributions the global research for novel and effective therapeutic strategies for multiple disease categories.     

Effects of Micronutrient Deficiencies on Human Health: Its Status in South Asia

SUMMARY

Vitamin A, iodine, and iron deficiencies affect large numbers of people worldwide. A deficiency of one or more of these micronutrients adversely affects the physical and mental abilities of humans. South Asia contains a high percentage of these individuals, not just because of its large population, but also the prevalence of deficiency is higher there than in many other parts of the world. The reasons for the high rates of deficiency in South Asia are many, but they include inadequate availability of micronutrient-rich foods, sub-optimal dietary habits, and high rates of infection. Across the region, national plans vary from (a) increasing availability of appropriate foods, (b) introducing new dietary behaviors, (c) improving health status, and (d) directly addressing deficiency through intervention programs. The extent to which each of the countries is making progress toward the goals of these plans can help in understanding the priorities for future efforts to address micronutrient malnutrition. This article discusses the adverse effects of micronutrient deficiencies in humans with special reference to South Asia.     

稻文化的再思考3:稻与社稷——印度、泰国等东南亚、南亚主要国家

东南亚、南亚地区是世界上重要的稻米产区,稻米在民众饮食生活和国家社会政治中占有重要地位,民众生活中也保存了非常悠久和丰富的稻作文化习俗及稻谷起源神话传说。本文分析了水稻在东南亚、南亚主要国家印度、印度尼西亚、泰国、越南、菲律宾、孟加拉国、缅甸的作用、地位、生产、消费、文化等情况,并指出了印度、印度尼西亚稻文化的主要标志。     

Irrigation but not N fertilization enhances seedhead density in plains rough fescue ( F estuca hallii)

Plains rough fescue ( F estuca hallii) is an important forage grass species in western Canada. Seed for use in pastures and ecological restoration is in high demand but supply is limited because F. hallii is an erratic seed producer. Seed producers require an understanding of the factors that influence flowering and seed set in this species. The aim of this study was to assess the effect of irrigation and nitrogen fertilization on the flowering and growth of F. hallii in a native rough fescue prairie in Alberta, Canada. Irrigation had a strong positive effect on seedhead density, whereas fertilization had limited effects on growth and reproduction of F. hallii. These results demonstrate that under field conditions, available moisture is likely to be a key factor driving flowering and seed production in F. hallii.     

Management of Soil Micronutrient Deficiencies in the Rice-Wheat Cropping System

Summary

Within the last three decades, the rice-wheat cropping system has triggered, and with time, aggravated soil micronutrient deficiencies in the Indo-Gangetic Plains (IGP). This has largely been due to the shift from an earlier rice and wheat monoculture with low yielding, long duration indigenous varieties, to an intensive rice-wheat rotation cropping system with short duration modern high yielding varieties on the same piece of land. The problems related to micronutrient deficiency in the IGP are more due to the size of its available pools in the soil rather than its total contents and are greatly influenced by crop management, or rather its mismanagement. Deficiency of zinc is widespread in the IGP, but with the extensive use of zinc sulfate, zinc deficiency has reduced in some areas of the region. Meanwhile, the deficiency of Fe, Mn and B has increased in the IGP. Deficiency of Cu and Mo is location specific and can limit rice and wheat yields. The adoption and spread of the rice-wheat system in permeable coarse textured soils, particularly in the western IGP, not only caused iron deficiency in rice but also resulted in the emergence of manganese deficiency in wheat. In highly calcareous and acidic soils, boron is the next limiting micronutrient in crop production after zinc. Bumper rice and wheat harvests in the past decade, the declining use of organic manures in the region and except for the widespread use of zinc sulfate, a general lack of awareness amongst farmers on micronutrient deficiency problems has contributed to micronutrients limiting rice and wheat yields in the IGP. Approaches to alleviating micronutrient deficiencies include matching the crop removals of the micronutrients with its replenishments through their respective external carriers, supplementation through organic sources and mobilization/utilization through cultivation of micronutrient efficient crop cultivars. Identification of efficient micronutrient carriers and finding the optimum rate, mode and time of its application is important in ameliorating the micronutrient deficiencies. This article reviews the extent of micronutrient deficiency and discusses various management options available to reduce micronutrient deficiency induced crop yield reduction for rice and wheat in the Indo-Gangetic Plains.     

Model of wheat yield response to application of diclofop-methyl to control ryegrass (Lolium rigidum)

A general model of crop yield response to herbicide application is proposed. The model includes three components: the effect of herbicide dosage on weed density, the effect of surviving weed density on crop yield and the effect of herbicide directly on the crop. The model is used to estimate the response of wheat yield to application of diclofop-methyl to control ryegrass (Lolium rigidum) in Australia. It is found that the competitiveness of ryegrass plants surviving treatment is reduced by the treatment and that the proportion of yield loss at a given ryegrass density is not independent of the absolute weed-free yield. The response function is used to calculate economic thresholds and optimal herbicide dosages.