辣椒环斑病毒分子检测方法的建立及应用

辣椒环斑病毒（Chilli ringspot virus, ChiRSV）是2007年在辣椒上发现的病毒新种,是马铃薯Y病毒属（Potyvirus）的确定种。笔者于2009年首次在海南黄灯笼辣椒上检测发现ChiRSV。根据ChiRSV保守区域设计和筛选PCR特异引物,优化退火温度,对PCR产物进行序列测定,建立了ChiRSV的RT-PCR检测方法。通过灵敏度测定,结果表明该方法最低可检出12.5 pg病毒RNA,灵敏度高。通过对来自海南田间的疑似辣椒病样的RT-PCR检测,证实了该方法有良好的反应特异性,亦表明ChiRSV可能已在海南扩散。     

黄灯笼辣椒组织培养中防褐变的研究

以黄灯笼辣椒无菌苗子叶为外植体,对其分化生长过程中的褐变现象进行研究。结果表明,通过在子叶不定芽分化培养基中添加0.2%活性炭(AC)、6mg/L硝酸银(AgNO3)和40mg/L维生素C(VC),均能不同程度的减轻褐变的程度,其中以6mg/LAgNO3效果最佳。     

黄灯笼辣椒种质资源遗传多样性ISSR分析

采用ISSR分子标记技术对22份国内外黄灯笼辣椒种质的遗传多样性进行了分析,为选配黄灯笼辣椒杂交育种亲本选择提供了参考.根据加拿大哥伦比亚大学公布的100对ISSR引物,从中筛选出17条多态性好、条带稳定的引物,用其对22份黄灯笼辣椒种质进行扩增,共获得154条谱带,其中140条为多态性条带,多态性比率为90.9％,表明ISSR标记对黄灯笼辣椒具有较高的多态性;通过对黄灯笼辣椒ISSR遗传相似系数的统计与分析,品种间的遗传相似系数在0.461～0.870之间,平均为0.778.聚类分析表明,22份材料在遗传相似系数0.52处分为2类,其中L508为种间杂种后代,与其它种质具有较远的亲缘关系,单独为一类;在遗传相似系数0.72处可以分为4类,把不同亲缘关系的种质明显区分开来,表明ISSR分子标记进行黄灯笼辣椒遗传多样性的分析是可行的.黄灯笼辣椒品种间具有较丰富的遗传多样性,并为黄灯笼辣椒品种的选育和种质资源的保护提供了理论依据.     

黄灯笼辣椒核心种质资源比较构建研究

核心种质的构建为种质资源的研究和有效利用提供了便利条件。以146份黄灯笼辣椒种质资源为试验材料,基于10个性状表型数据,采用混合线性模型分析方法无偏地预测基因型值,利用马氏距离计算种质间的遗传距离,分别采用2种取样方法（随机取样法和优先取样法）,2种聚类方法（离差平方和法和类平均法）,按照30％的抽样比率构建黄灯笼辣椒核心种质库。采用均值、方差、极差和变异系数4个指标评价不同取样方法和聚类方法构建核心种质库的优劣。试验结果表明,优先取样法优于随机取样法,类平均法优于离差平方和法。基于马氏距离、优先取样法、类平均法获取的43份黄灯笼椒核心资源能够代表原群体的遗传多样性。     

砂培黄灯笼辣椒根系分泌物化学成分鉴定

鉴定黄灯笼辣椒根系分泌物的化学成分,为研究其化感作用及连作障碍的机理提供理论依据。采用砂培法收集黄灯笼辣椒的根系分泌物,柱层析法分离根系分泌物,依次选用石油醚、乙酸乙酯、氯仿和甲醇洗脱根系分泌物并收集各洗脱组分,采用气质联用仪(GC-MS)鉴定各组分的化学成分。研究结果表明,黄灯笼辣椒根系分泌物石油醚组分GC-MS鉴定图谱共出现57个物质峰,相似度大于85%的化学物质共28个;乙酸乙酯组分GC-MS鉴定图谱共出现73个物质峰,相似度大于85%的化学物质共28个;氯仿组分GC-MS鉴定图谱共出现67个物质峰,相似度大于85%的化学物质共27个;甲醇组分GC-MS鉴定图谱共出现55个物质峰,相似度大于85%的化学物质共28个。黄灯笼辣椒根系分泌物中含有烷烃类、脂类、酸类、醇类、酚类、醛类、肟类、吡咯烷酮类、酰胺类和萘胺类化合物。     

海南黄灯笼辣椒小叶病病原物的分子鉴定

采用分子生物学技术对田间疑似植原体侵染引起的小叶症状的黄灯笼辣椒样品进行检测,以明确其病原分类地位。利用植原体通用引物对R16mF2/R16mR1和R16F2n/R16R2,对上述黄灯笼辣椒叶片DNA进行巢式PCR扩增。结果表明:获得约1.2 kb的特异片段,经序列测定、Blast同源性检索及进化树和iPhyClassifier分析,获得该特异片段长为1 248 bp,与植原体16S rⅡ组中的花生丛枝植原体(L33765)同源性最高,达99%,聚类在同一个进化枝上,相似性系数为1.00,归属于16S rII-A亚组。暂将其命名为黄灯笼辣椒小叶植原体(Capsicum chinense Jacquin little-leaf disease phytoplasma,CcJLL)。     

DA-6对高温胁迫下黄灯笼辣椒幼苗的影响

以海南黄灯笼辣椒"热辣2号"为试验材料,分别对幼苗喷施10、20、30、40、50 mg/L的DA-6,再进行高温胁迫处理,研究不同浓度的DA-6对缓解黄灯笼辣椒幼苗高温胁迫的作用。结果表明:喷施DA-6的辣椒幼苗各项生理指标较对照存在显著差异;喷施30 mg/L的DA-6时,幼苗的叶色值、根系活力、MDA含量、SOD活性和脯氨酸含量等指标综合反应较好,与其他处理差异显著。高温胁迫下,DA-6可有效增强黄灯笼辣椒幼苗的光合能力,喷施30 mg/L DA-6的辣椒幼苗净光合速率(Pn)、气孔导度(Gs)和蒸腾速率(Tr)与对照相比均有所提高,分别提高了0.46、1.69、1.37倍;其PSⅡ最大荧光效率(Fv/Fm)、PSⅡ量子产量(Yield)、非光化学淬灭(qN)与光化学淬灭(qP)均显著高于对照。因此,DA-6能够缓解高温胁迫对黄灯笼辣椒幼苗的影响,其最适宜浓度为30 mg/L。     

海南黄灯笼辣椒顶死病病原病毒的分离鉴定

从海南省黄灯笼辣椒顶死病株上分离纯化得到一个病毒分离物.研究结果表明,该病毒分离物能通过汁液磨擦接种侵染供试植物中的4科11种植物,可由桃蚜传播;提纯病毒粒子呈球形,直径28～30 nm,外壳蛋白分子量约为28 ku;分离物与CMV抗血清在ELISA测定中呈阳性反应;提取病毒分离物RNA,应用RT-PCR方法克隆了病毒外壳蛋白（CP）基因.CP基因675 bp,编码218个氨基酸.对CP基因序列分析表明,该病毒分离物的CP基因核苷酸序列与黄瓜花叶病毒亚组Ⅰ分离物的同源性均在92.1%以上,所推导的氨基酸序列同源性在96.3%以上,而与亚组Ⅱ分离物的核苷酸序列同源性均低于77.3%,所推导的氨基酸序列同源性均低于80.7%.据此将该病毒分离物鉴定为黄瓜花叶病毒,归属于亚组Ⅰ.     

姜油树脂的超临界CO_2萃取条件及其抑菌活性

采用正交试验,优化了超临界CO2萃取姜油树脂的工艺条件,在压力22 Mpa,时间15 min,温度50℃,萃取率可达5.38%。抑菌试验结果表明,姜油树脂对大肠杆菌、沙门氏菌、痢疾志贺菌、苏云金杆菌和金黄色葡萄球菌均有一定的抑制作用,抑菌最低质量浓度为0.5～1.0 mg/mL。在不同温度或不同时间紫外光照条件下,姜油树脂仍有一定的抑菌作用,表明其具有一定的光、热稳定性。     

金线莲乙醇提取物对金黄色葡萄球菌抑菌性能研究

用乙醇作为溶剂,提取出金线莲中的有效抑菌物质,采用二倍稀释法确定金线莲乙醇提取物对金黄色葡萄球菌的最小抑菌浓度(Minimal inhibition concentration,MIC)为5.000 mg/m L;并经介质pH、热处理和紫外线处理提取物,发现其抑菌圈直径在pH9.0和30℃下最大;分析金线莲乙醇提取物对供试菌蛋白质和还原糖的利用能力,表明金线莲乙醇提取物通过减弱蛋白质和还原糖的代谢来抑制细菌的生长繁殖。     

Chemical Composition and Antioxidant/Antimicrobial Activities in Supercritical Carbon Dioxide Fluid Extract of Gloiopeltis tenax

Gloiopeltis tenax (G. tenax) is widely distributed along the Chinese coastal areas and is commonly used in the treatment of diarrhea and colitis. This study aimed at investigating the bioactivities of the volatile constituents in G. tenax. We extracted the essential constituents of G. tenax by supercritical carbon dioxide extraction (CO₂-SFE), then identified and analyzed the constituents by gas chromatography-mass spectrometry (GC-MS). In total, 30 components were identified in the G. tenax extract. The components showed remarkable antioxidant activity (radical scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH)), lipid peroxidation inhibition capacity (in a β-carotene/linoleic acid-coupled oxidation reaction), and hydroxyl radical-scavenging activity (by deoxyribose degradation by iron-dependent hydroxyl radical), compared to butylated hydroxytoluene. In microdilution assays, G. tenax extracts showed a moderate inhibitory effects on Staphyloccocus aureus (minimum inhibitory concentration (MIC) = 3.9 mg/mL), Enterococcus faecalis (7.8 mg/mL), Pseudomonas aeruginosa (15.6 mg/mL), and Escherichia coli (3.9 mg/mL). Antioxidant and antimicrobial activities of G. tenax were related to the active chemical composition. These results suggest that the CO₂-SFE extract from G. tenax has potential to be used as a natural antioxidant and antimicrobial agent in food processing.     

超临界法萃取的黑胡椒油树脂成分分析

在超临界CO2萃取压力为30 Mpa、萃取时间为3 h、萃取温度为50℃的静态条件下,分别萃取未做任何处理、无水乙醇处理、乙酸乙酯处理的黑胡椒原料中的油树脂,并利用气相色谱-质谱联用(GC-MS)对萃取得到的黑胡椒油树脂进行成分分析,同时利用保留指数和MS对物质进行定性分析。结果显示,在超临界萃取条件下,未做任何前处理的黑胡椒油树脂的香气成分有22种,采用无水乙醇进行前处理的黑胡椒油树脂的香气成分有17种,采用乙酸乙酯进行前处理的黑胡椒油树脂的香气成分有41种。     

二氧化碳浓度升高对茶树生理代谢的影响及应对技术

大气中的二氧化碳(CO2)浓度升高是工业革命以来全球范围内最重要的生态变化之一,将直接影响植物的生长发育和代谢过程。尽管CO2浓度升高对粮食作物的影响已经得到了广泛的研究,但其对茶树等重要经济作物的影响却很少受到关注。本文回顾和总结了CO2浓度升高对茶树的初级代谢(包括光合作用、呼吸作用和碳氮代谢)和次级代谢的影响,并探讨了CO2浓度升高环境下茶叶生产过程中的应对技术,旨在为CO2浓度升高背景下茶树优质高产栽培提供一定的理论基础和科学依据。     

Evapotranspiration Responses of Plants and Crops to Carbon Dioxide and Temperature

Summary

Atmospheric carbon dioxide (CO2) concentration has risen from about 270 mmol (CO2) mol^?1 (air) (i.e., mole fraction of dry atmospheric air basis) before 1700 to about 370 mmol mol ^?1 currently. General Circulation Models (GCM) have predicted a global temperature rise of 2.8 to 5.2°C for a doubling of CO2. This review examines evapotranspiration and water-use efficiency responses of plants to rising CO2 and climatic changes, especially temperature. Doubling of CO2 will decrease leaf stomatal conductance to water vapor about 40%. However, water use by C3 crop plants under field conditions has usually been decreased only 12% or less for two reasons. Firstly, feedbacks in the energy balance of plant foliage cause leaf temperatures to rise as stomatal conductance is decreased. Increases of leaf temperature raise the vapor pressure of water inside the leaf, which increases the leaf-to-air vapor pressure difference. This increased driving force for transpiration offsets in large part the decreased leaf conductance caused by elevated CO2. Secondly, CO2 enrichment tends to cause leaf area to increase more rapidly in many crops. This increased leaf surface area for transpiration also offsets part of the decreased stomatal conductance per unit leaf area on the whole canopy evapotranspiration, but the energy budget feedbacks are more important.

Experiments point to a yield enhancement of 30 to 35% for C3 crops for the direct effects a doubling of CO2 (without ancillary climate change). If temperature rises, this yield enhancement may be greater for vegetative growth but less for seed grain yield. Experiments on both ambient and elevated CO2 treatments in sunlit growth chambers showed that transpiration rates increased 20% when air temperature was changed from 28 to 33 °C and increased 30% when temperature was increased from 28 to 35 °C. Thus, under well-watered conditions, evapotranspiration will increase about 4 to 5% per 1°C rise in temperature.

Crop model predictions of yields of soybean and maize showed a reduction due to temperature increases by two GCM models. Under Southeastern USA conditions, doubling CO2 in the Goddard Institute for Space Studies (GISS) climate change scenario resulted in an 12% increase in yields, but yields decreased 50% in the Geophysical Fluids Dynamics Laboratory (GFDL) climate change scenario. Optimum irrigation for both models gave yield increases of about 10%. These model results illustrate the critical requirement of water for production of crops. Under rainfed conditions, crop yields could suffer tremendously if growing season precipitation is decreased, but yields could increase moderately if growing season precipitation is increased. Under the high growing season rainfall scenario (GISS), irrigation requirements for optimum soil water were increased 22%, but under the low rainfall scenario (GFDL), irrigation requirements were increased 111%.

Without the effects of climate change, rising CO2 will cause an increase in crop water-use efficiency (WUE). Most of the increases in WUE will be due to increases in dry matter, with little or no contribution from decreases in water use per unit land area. Growers could produce higher yields per unit land area with higher total production, or maintain the same total production with less land and less total water use. However, if temperatures rise, transpirational water use will increase, and WUE will decline. Higher temperatures, and especially less rainfall, would raise the irrigation requirements of crops. Competition for water resources from other uses could result in less water available for irrigation.     

CO2浓度升高条件下水稻蒸腾与N吸收的关系

利用FACE（Free Air Carbon Dioxide Enrichment）平台技术,用水培试验研究了低氮（14 mg/L）和高氮（28 mg/L）水平下,大气CO2浓度升高条件下水稻蒸腾与N吸收速率的相关关系。结果表明,在CO2浓度升高条件下,水稻生物量增加了36%（低N）和29%（高N）;总吸N量也增加达7%（低N） 和5%（高N）;而总蒸腾量减少28%（低N）和10%（高N）。由于促进更多分蘖的发生,高CO2浓度使分蘖期水稻平均N吸收速率提高了31%～156%（低N）和19%～87%（高N）,在其他时期无明显影响;而高CO2浓度对水稻平均蒸腾速率的影响主要表现在抽穗到灌浆末期。在对照条件下,平均蒸腾速率和平均N吸收速率呈显著正相关;但在CO2浓度升高条件下,两者相关关系不显著。说明人们所推测的“蒸腾效应”——高CO2浓度条件下降低了的蒸腾作用并非影响水稻N吸收的关键因素。     

玉米氮、磷、钾、水和二氧化碳资源积累与利用的Meta分析

按照Meta分析原理,汇集近年来的玉米田间试验数据共722组,进行稳健统计、梯度和等值线分析。氮、磷、钾、水和二氧化碳共5个数据集的玉米产量中位数变化在7.91~10.61 t/hm2之间,生产百公斤玉米子粒N、P2O5、K2O、CO2、H2O的资源需求量分别为1.98、0.85、2.12、298.8 kg和44.2 m3。产量偏导数比值(资源利用效率的偏导数/积累量的偏导数)有3种表现,水与氮归为一类,产量偏导数比值大多数都在1/2以下,少数值为1,说明在大多数情况下,积累量的增产作用是资源利用效率的2倍以上,少数情况下二者作用相当;钾与磷为一类,产量偏导数比值以1最多,1/2和2次之,说明以资源利用效率和积累量的作用相当为主,但是,也有不少利用效率的作用是积累量的2倍以上,或者相反;二氧化碳绝大多数产量偏导数比值在2以上,较少部分的值为1,说明在绝大多数情况下资源利用效率的增产作用是积累量的2倍以上,少数情况下二者作用相当。当代玉米百公斤子粒的资源需求量明显下降,资源积累(消耗)量和利用效率对产量的相对重要性因二者组合所处的区域而异,不同资源因子对产量的作用格局不同。     

Biomedical exploitation of chitin and chitosan via mechano-chemical disassembly, electrospinning, dissolution in imidazolium ionic liquids, and supercritical drying

Recently developed technology permits to optimize simultaneously surface area, porosity, density, rigidity and surface morphology of chitin-derived materials of biomedical interest. Safe and ecofriendly disassembly of chitin has superseded the dangerous acid hydrolysis and provides higher yields and scaling-up possibilities: the chitosan nanofibrils are finding applications in reinforced bone scaffolds and composite dressings for dermal wounds. Electrospun chitosan nanofibers, in the form of biocompatible thin mats and non-wovens, are being actively studied: composites of gelatin + chitosan + polyurethane have been proposed for cardiac valves and for nerve conduits; fibers are also manufactured from electrospun particles that self-assemble during subsequent freeze-drying. Ionic liquids (salts of alkylated imidazolium) are suitable as non-aqueous solvents that permit desirable reactions to occur for drug delivery purposes. Gel drying with supercritical CO(2) leads to structures most similar to the extracellular matrix, even when the chitosan is crosslinked, or in combination with metal oxides of interest in orthopedics.     

试论大气CO2浓度升高条件下水稻碳氮代谢变化及其调控途径

 以水稻为例，从分析高等植物光合速率对大气CO2浓度升高的短期与长期反应入手，结合水稻的氮素营养特性，通过分析大气CO2浓度倍增下，水稻体内氮素营养状况与光合速率的关系，以及光合作用过程中酶的活性与总量、光合底物的再生与水稻氮素营养的关系，论述了大气CO2浓度升高对水稻体内碳氮代谢的影响。对稻田生态系统的氮素营养管理提出了建议，并从产量和生态角度对今后水稻氮素营养的研究进行了展望。     

Supercritical carbon dioxide (SC-CO2) extraction of Nigella sativa L. oil using full factorial design

In this study, Nigella sativa L. oil was extracted using supercritical carbon dioxide with full factorial design to determine the best extraction condition (pressure, temperature and dynamic extraction time) for obtaining an extract with high yield, antioxidant activity and thymoquinone (TQ) quantity. The maximum thymoquinone content in the highest overall yield was achieved through SC-CO₂ extraction condition of 150 bar, 40 °C, 120 min with the value of 4.09 mg/ml. The highest SC-CO₂ extraction yield was 23.20% which obtained through extraction condition of 350 bar, 60 °C and 120 min. The extraction conducted at 350 bar, 50 °C, 60 min showed the lowest IC₅₀ value (highest antioxidant activity) of 2.59 mg/ml using DPPH radical scavenging activity method. Fatty acid composition of the extracted oil with highest radical scavenging activity was obtained by gas chromatographic analysis.     

响应面法制备枇杷核微孔炭及其选择性吸附研究

以新鲜枇杷核为原料制备了枇杷核微孔炭（LMC）,以 LMC 对盐酸四环素的吸附率为指标,通过响应面法优化制备工艺,进一步研究了 LMC 的孔结构和选择性吸附性能。结果表明：最优制备工艺为活化时间 192 min、活化温度 565 ℃、活化升温速率 21.5 ℃/min、炭化温度 632 ℃;该工艺所得枇杷核微孔炭对盐酸四环素的吸附率高达 95.25%,其 BET 比表面积为 1 275 m2/g,微孔表面积为 1 219 m2/g,总孔容为 0.535 4 cm3/g,微孔孔容为0.473 cm3/g;较高的比表面积和高比例的微孔含量赋予其优异的分子尺寸选择性吸附性能。