Antioxidant activity and phenolic content of chestnut (Castanea sativa) shell and eucalyptus (Eucalyptus globulus) bark extracts

Chestnut (Castanea sativa) shell and eucalyptus (Eucalyptus globulus) bark, waste products of the food and wood industries, respectively, were analysed as potential sources of antioxidant compounds. The extraction yield, the antioxidant activity and total phenols content of the extracts were greater in chestnut shell than in eucalyptus bark for most of the extraction conditions essayed. Extraction of chestnut shell with a 2.5% Na₂SO₃ aqueous solution led to the highest extraction yield, 25.6%, total phenols, 13.4 g gallic acid equivalent/100 g oven-dried shell, and FRAP antioxidant activity, 80.7 mmol ascorbic acid equivalent/100 g oven-dried shell. Extraction with methanol:water (50:50, v/v) provided the best results for eucalyptus bark. The antioxidant activity and the total phenols content of the extracts had a positive linear correlation. FTIR spectroscopy confirmed the higher content of phenolic compounds in chestnut shell extracts compared to eucalyptus bark extracts. Chestnut shell extracts were characterized by the presence of high molecular weight species whereas lower molecular weight species were predominant in eucalyptus bark extracts.     

Antiradical properties of sorghum (Sorghum bicolor L. Moench) flour extracts

Epidemiological studies support the belief that whole grains are protective against several chronic diseases. The health benefits of whole grains are attributed in part to their unique phytochemical composition. Major phytochemicals in grains include various classes of phenolic compounds, flavonoids and coumarin derivatives, etc. Phenolic compounds present in grains possess antioxidant properties that are associated with the health benefits of grains and grain products. Sorghum is one of the main staple cereal grains in hot dry tropics and ranks fifth among cereal crops in the world. Although sorghum is rich in phenolics and tannins which are proven anticancer and cardioprotective constituents, human consumption of sorghum is limited. To our knowledge, there is limited literature on the profile of antioxidant phytochemicals in the local white variety of sorghum. Hence, the objective of this study was to investigate the antioxidant property of white sorghum flour extracts in vitro and also to identify the fractions responsible for the antioxidant activity. In the present study, we analyzed the antioxidative properties of various extracts (water, 60% methanol, 60% ethanol, and 60% t-butanol) of white sorghum flour employing the 1,1-diphenyl-2-picrylhydrazyl (DPPH) model system. Phenolics, antiradical and antioxidant activities were also examined in chromatographic sub-fractions of the soxhlet methanolic extract. Our results indicated that the various extracts exhibited significant antioxidant activity that did not correlate with the phenolic content. Further, two sub-fractions eluted with methanol and acetone/methanol were found to possess strong antioxidant activity in two assay systems. Our results suggest that a diet rich in sorghum may be useful in combating diseases in which free radical production plays a key role.     

Potential use of Asteraceae extracts to control Spodoptera frugiperda (Lepidoptera: Noctuidae) and selectivity to their parasitoids Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) and Telenomus remus (Hymenoptera: Scelionidae)

Extracts of 12 Asteraceae were tested on Spodoptera frugiperda (Lepidoptera: Noctuidae) and on their parasitoids Telenomus remus (Hymenoptera: Scelionidae) and Trichogramma pretiosum (Hymenoptera: Trichogrammatidae). The plants Lychnophora ericoides and Trichogonia villosa were toxic for 97.7 ± 0.15% of one-day-old eggs of S. frugiperda and Lepidaploa lilacina for 72.0 ± 2.50% for two-day-old eggs of this insect. Extracts of Vernonia holosenicea, Lychnophora ramosissima and Chromolaena chaseae had higher impact on S. frugiperda, while those of Eremanthus elaeagnus and L. ericoides were more selective to T. pretiosum and T. remus. Asteraceae extracts present potential for integrated pest management programs of S. frugiperda.     

Chemical composition and antioxidant and antimicrobial activity of essential oil of Artemisia annua L. from Bosnia

Hydrodistilled volatile oil obtained from the aerial parts of Artemisia annua L., cultivated near Sarajevo, Bosnia, was analyzed by GC-MS. More than one hundred compounds were identi?ed, representing 95.5% of the total oil. The major constituents of essential oil were oxygenated monoterpenes, artemisia ketone (30.7%) and camphor (15.8%). Isolated essential oil was tested for radical-scavenging ability using the stable DPPH radical, the ABTS radical, for reducing power ability with a test based on the reduction of ferric cations, for reducing ability of hydroxy radical in ORAC assay, and for metal chelating ability using the ferrozine assay. In all tests oil did not show a prominent antioxidant activity, but still comparable with thymol, an already known antioxidant. The screening of antimicrobial activity of oil was individually evaluated against representatives of Gram-positive, Gram-negative bacteria and fungi, using the agar diffusion method. All tested microorganisms were inhibited by essential oil. To the best of our knowledge, this is the first report of antimicrobial activity of essential oil of A. annua against Haemophilus influenzae, Enterococcus faecalis, Streptococcus pneumoniae, Micrococcus luteus and Candida krusei microbial strains. The antioxidant, antibacterial and antifungal activity of essential oil of A. annua from Bosnia is presented here for the first time and extends our knowledge in the range of valuable biological activities and possible roles in therapy associated with this medicinal herb.     

Composition and physical properties of cress (Lepidium sativum L.) and field pennycress (Thlaspi arvense L.) oils

The fatty acid profiles and tocopherol and phytosterol contents of crude oils of cress (Lepidium sativum L.) and field pennycress (Thlaspi arvense L.) are reported, along with yields from the corresponding seeds. The physical properties of these oils were also determined, which included oxidative stability, kinematic viscosity, viscosity index, low temperature fluidity, specific gravity, acid value, lubricity, and iodine value. The oil content of dried cress and field pennycress seeds was 22.7 and 29.0 wt%, respectively. The primary fatty acids found in cress oil were oleic (30.6 wt%) and linolenic acids (29.3 wt%), whereas field pennycress oil was principally composed of erucic (32.8 wt%) and linoleic (22.4 wt%) acids. Cress oil contained high concentrations of γ- (1422 ppm) and δ- (356 ppm) tocopherols, whereas α-tocopherol (714 ppm) was the primary tocopherol discovered in field pennycress oil. The overall tocopherol concentrations of cress and field pennycress oils were 1799 and 851 ppm, respectively. The primary phytosterols elucidated in cress and field pennycress oils were sitosterol and campesterol, with avenasterol also present in significant quantity in cress oil. The total phytosterol concentration in cress oil (14.41 mg/g) was greater than that in field pennycress (8.55 mg/g) oil. Field pennycress oil exhibited excellent low temperature fluidity, whereas cress oil was more stable to oxidation and over a range of temperatures displayed lower kinematic viscosities as well as a higher viscosity index. The acid and iodine values of field pennycress oil were lower than those for cress oil, but both oils had excellent lubrication properties.     

小麦丙氨酸-乙醛酸转氨酶基因（ TaAGT2）的克隆、生物信息学预测及表达分析

光呼吸是植物细胞H_2O_2的重要来源,也是维持细胞氧化还原的重要成分,影响植物对生物和非生物逆境的应答,因此,挖掘与光呼吸有关的基因对提高植物的抗逆性具有重要意义。本研究以耐旱小麦品种青麦6号转录组数据为基础,通过RACE技术克隆得到小麦 AGT2基因的全长cDNA,将其命名为 TaAGT2,并对其进行了生物信息学预测及表达模式分析。结果表明, TaAGT2开放阅读框长1 434bp,编码477个氨基酸。该蛋白属于亲水性稳定蛋白,定位于线粒体中,二级结构以α-螺旋(42.14%)和无规则卷曲(32.91%)为主。 TaAGT2基因包含AAT-I基因族保守区域,与二穗短柄草 AGT2的相似性高达97%。通过qRT-PCR对基因 TaAGT2在不同组织中(根、茎和叶)及4种非生物胁迫(低温、ABA、干旱、高盐)下的表达特性进行分析,结果表明, TaAGT2在根、茎和叶中均有表达,茎中的表达显著高于根和叶,在不同胁迫条件下上调表达,表明该基因可能参与调控植物的抗逆反应。     

Enzyme inhibitory and antioxidant activity of Melia azedarach L. naturalized in Anatolia and its phenolic acid and fatty acid composition

In the present study, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and tyrosinase inhibitory and antioxidant activities of the fruit and leaf extracts of Melia azedarach L. (Meliaceae) of Turkish origin were evaluated. Enzyme inhibitory activity of the extracts was tested in vitro using ELISA microplate reader. Antioxidant activity of the extracts was tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferrous ion-chelation, and ferric-reducing antioxidant power (FRAP) assays. Phenolic composition of the extracts was elucidated by high performance liquid chromatography (HPLC) and fatty acid compositions of the fatty oils of the fruits and leaves were elucidated by gas chromatography-mass spectrometry (GC-MS). The ethyl acetate extract from the leaves showed the highest inhibition against AChE (33.63 ± 1.40%) and BChE (92.89 ± 3.05%). The methanol extract from the leaves exerted the best antioxidant activity in DPPH radical scavenging and FRAP assays, while the ethyl acetate extracts of the fruits and leaves had the most notable effect in metal-chelation assay.     

玉米脂肪酸α-双加氧酶cDNA克隆与表达分析

以玉米自交系丹340为实验材料,同源克隆到玉米脂肪酸α-双加氧酶基因(ZmDOX)cDNA序列,并利用半定量与定量RT-PCR分析其在不同组织和PEG胁迫下的表达特征。序列分析结果表明,该cDNA序列包含1 857 bp完整的开放阅读框,编码619个氨基酸残基的蛋白产物。玉米、水稻、小麦中DOX氨基酸序列高度同源,聚集在同一进化枝,而双子叶植物存在两类DOX蛋白。组织表达特征分析发现,ZmDOX在根、茎、叶中均有表达,其中在根中表达量最高。PEG胁迫条件下,根中ZmDOX对胁迫的响应较快且增加幅度较大,而在叶中表达量有降低的趋势。     

普通小麦TaADF8基因的克隆及表达分析

肌动蛋白解聚合因子(actin-depolymerizing factor,ADF)普遍存在于真核细胞中,为低分子量的肌动蛋白结合蛋白,在调控细胞内肌动蛋白纤维的聚合和解聚中起关键作用。为给深入研究TaADF8基因在小麦中的功能机理奠定基础,并为进一步丰富小麦ADF基因研究内容提供理论参考,本研究利用电子克隆策略从小麦品种CP53中克隆出TaADF8基因(GenBank登录号为KJ864962)后对其进行序列分析,并进一步采用荧光定量PCR(quantitative real-time PCR,qRT-PCR)技术对其在小麦不同组织间的表达差异及不同非生物胁迫下的表达模式进行分析。核酸序列分析表明,该基因全长695bp,拥有完整的ORF,编码142个氨基酸。氨基酸序列分析表明,该蛋白含有保守的ADF同源区和PIP2结合结构域,且在氨基端有核定位信号。进化和聚类分析表明,小麦TaADF8基因与大麦HvADF2基因、HvADF3基因和水稻OsADF3基因亲缘关系较近,蛋白相似度分别为75.35%、93.66%和67.86%。qRT-PCR表达特性分析显示,该基因为组成型表达,在根、茎、叶、颖壳和雄蕊中均表达,且在根、叶和雄蕊中表达量较高;该基因表达受低温的强烈诱导,同时也受水分、高盐和外源脱落酸胁迫诱导。     

Field trials of Metarhizium anisopliae var. acridum (Ascomycota: Hypocreales) against oriental migratory locusts, Locusta migratoria manilensis (Meyen) in Northern China

During 2002–2006, nymph bands of Locusta migratoria manilensis (Meyen) were treated by ground and aerial applications in 6000 ha of grasslands and the nearby beach of Yellow river using a soybean oil miscible suspension ULV formulation of Metarhizium anisopliae var. acridum isolate CQMa102. The formulation was also applied in Tianjin, Henan, Hebei, Shandong and Shanxi provinces of Northern China by ground and aerial applications. During field studies, cage tests were carried out in corresponding field plots in order to estimate the mortality accurately. Doses of 3.3×10¹² and 5.0×10¹² conidia ha⁻¹ were equally effective and caused 90% mortality 9–13 days after treatment. In the ground spray trial, 3.3×10¹² conidia ha⁻¹ killed >90% of L. migratoria manilensis 11–15 days after treatment in a wide variety of vegetation and weather conditions. The decline of locust populations was slower where vegetation was taller and denser. In the aerial spray treatment, the final percent survival of locusts was lowered to 10% at 11 and 14 days in the field cage and open field locusts, respectively. Furthermore, the M. anisopliae oil miscible suspension formulation did not appear to harm natural enemies of locusts in the field.     

白僵菌与苏云金芽胞杆菌水悬浮剂研制及田间防治玉米螟研究

通过室内生物测定,研究球孢白僵菌和苏云金芽胞杆菌联合防治玉米螟的效果。研究表明,确定白僵菌与苏云金芽胞杆菌浓度分别为1.0×106孢子/mL、1.0×109孢子/mL条件下,杀虫效果达到最佳,LT90为6.64(±0.07)d,较1.0×109的Bb LT90缩短26.06%。通过助剂悬浮率、乳化效果以及助剂对微生物活性影响等确定1个白僵菌与苏云金芽孢杆菌水悬浮剂剂型配方。田间防治玉米螟结果表明,两种生防菌均在防治过程中起作用,田间玉米受害株数、活虫数均明显低于未施药对照,防治效果优于单一使用白僵菌或苏云金芽胞杆菌。     

小麦L699叶片受白粉菌胁迫后蛋白质的差异表达

为从蛋白组学角度认识小麦抗白粉病机制,以含有抗白粉病新基因Pm40的小麦品系L699为材料,采用蛋白质双向电泳和质谱技术(MALDI-TOF-MS)检测小麦叶片接种白粉菌24h后的差异蛋白。结果表明,经PDQuest软件分析,接种白粉菌后小麦叶片中有18个蛋白质斑点表达量上调,38个蛋白质斑点表达量下调。对表达量上调的蛋白质斑点进行质谱分析和数据库检索,共鉴定出15种蛋白质,其中13种参与防御反应、碳水化合物代谢和蛋白质周转。这些差异蛋白与抗病途径、植株生理过程密切相关,在小麦抗白粉病过程中起很重要的作用。     

昆虫病原线虫共生菌对小麦根腐病菌和小麦赤霉病菌的抑菌活性

为探索小麦根腐病和小麦赤霉病可持续控制的有效生防途径,以小麦根腐病菌和小麦赤霉病菌作为供试真菌,评价了昆虫病原线虫共生菌及其毒素的抑菌活性,并对高毒力菌株抗逆性进行了初探。结果表明,供试的28个昆虫病原线虫共生菌发酵液和毒素均有一定的抑菌活性,其中,高毒力共生菌菌株SY5发酵液和毒素对小麦根腐病菌的抑菌率分别为56.05%和67.41%,对小麦赤霉病菌的抑菌率分别为82.41%和83.32%;菌株SY5发酵液经50℃处理60 min及18 W紫外灯照射120 min后对小麦根腐病菌的抑菌率无明显变化,但对小麦赤霉病菌的抑菌率有所下降;常温保存150 d,抑菌活性略有下降。说明共生菌菌株SY5对小麦根腐病菌和小麦赤霉病菌抑菌活性显著,且具有一定的抗逆性,极具应用潜力。     

农杆菌介导的小麦成熟胚遗传转化影响因素分析

为解决农杆菌介导的小麦成熟胚遗传转化效率较低的难题,以春小麦Bobwhite成熟胚为外植体,分析植物激素、愈伤组织预培养时间、侵染时间和共培养阶段干燥处理对小麦成熟胚愈伤组织遗传转化的影响,并在遗传转化处理中辅以3个组织再生相关基因(TaSERK1、TaSERK2、TaNiR)的qRT-PCR分析。结果表明,愈伤组织预培养阶段添加适量的2,4-D和Picloram均可诱导成熟胚愈伤组织的形成,但是2mg·L~(-1)Picloram培养基中愈伤组织的胚萌发率为20.78%,远高于2,4-D诱导的胚萌发率(11.38%);共培养阶段对农杆菌侵染的愈伤组织进行干燥处理能适当提高愈伤组织的转化效率;qRT-PCR分析发现再生相关基因TaNiR相对表达量的增加能适当提高愈伤组织的转化率。     

Mechanisms Leading to Excess Alpha -Amylase Activity in Wheat (Triticum aestivum, L) Grain in the U.K.

The frequency and mechanisms of four modes of alpha -amylase enzyme accumulation in U.K. wheat, retained pericarp alpha -amylase activity (RPAA), pre-maturity alpha -amylase activity (PMAA), pre-maturity sprouting (PrMS) and post-maturity sprouting (PoMS), were investigated in field and laboratory experiments. Of 56 cultivar site year combinations (four model cultivars grown at up to four sites from 1994–1997), enzyme activity was detected in 32 cases, in 23 cases sufficient to reduce Hagberg falling number (the usual industry measure of alpha -amylase) below the commercial criterion (250 s). The frequency of occurrence of different modes of enzyme accumulation was in the order PoMS>PMAA>PrMS>RPAA. Both PMAA and PrMS were more common than expected and the most usual pattern was for alpha -amylase to accumulate by several modes. Although green grains are rejected as impurities, study of grain colour in relation to pericarp alpha -amylase activity showed that the enzyme could persist in non-green grains in levels sufficient to affect the Hagberg value. Two factors thought to promote PMAA, grain drying rate and transient changes in temperature in early development, were studied in the field and controlled environment cabinets. No significant difference was found in grain drying rate between samples where PMAA was or was not identified. However, out of 19 transfers from a cool (16/10 °C) to a warm (26/20 °C) temperature regime, six led to significant increases in PMAA. No transfers after 45% grain moisture increased PMAA. PrMS occurred as early as 67% grain moisture and susceptibility usually increased with stage of development, being greatest in the grain dough stage. PrMS susceptibility varied with cultivar (in the same order as PoMS sensitivity) and was affected by environmental factors.     

Chemical composition of the essential oil and antioxidant activity of methanolic extracts from fruits and flowers of Hypericum lydium Boiss.

This study was conducted to evaluate the essential oil composition and antioxidant activity of methanolic extracts from fruits and flowers of Hypericum lydium Boiss., a wild growing species of the Turkish flora. The antioxidant activities were determined through several biochemical assays. Both extracts showed an inhibitory effect against the formation of TBARS in a phosphatidylcholine liposome model system, moderate scavenging effect on DPPH and superoxide radicals, prominent reducing power and inhibitory effect on deoxyribose degradation in both the nonsite and site-specific assay, but a greater hydroxyl radical scavenging activity was observed in the non-site specific assay, suggesting that the extracts were better at scavenging hydroxyl radicals than at chelating iron. Correlation analysis indicated that there was a linear relationship between antioxidant potency, free-radical scavenging activity, reducing power and the content of flavonoids of fruits and flowers extracts. The essential oils obtained by hydrodistillation were analyzed by GC and GC-MS. In total forty-three compounds were identified. The most abundant components were monoterpenes hydrocarbons represented principally by α-pinene. The tested oil showed no antioxidant activity.     

小麦β-酮脂酰CoA合成酶基因KCS的克隆与酵母表达

超长链脂肪酸是生物体内众多重要物质的合成底物。KCS基因编码β-酮脂酰CoA合成酶,该酶具有底物特异性,参与超长链脂肪酸延伸的缩合反应,是超长链脂肪酸合成的限速步骤。为了探究小麦KCS基因在超长链脂肪酸合成中的功能,采用同源克隆的方法从小麦(Triticum aestivum L.)中克隆出KCS基因后,利用生物信息学对其编码序列进行分析,并在酿酒酵母(Saccharomyces cerevisiae)中对其进行真核表达。结果表明,小麦TaKCS6基因的开放阅读框为1 287bp,编码428个氨基酸残基。结构域预测结果显示,TaKCS6蛋白含有III型聚酮合酶脂肪酸延伸酶和C末端3-酰基ACP合酶III结构域,属于KCSs蛋白家族。序列比对分析结果显示,TaKCS6氨基酸序列与拟南芥及其他植物的KCS6氨基酸序列在两个功能结构域上和活性位点保守。酵母表达结果显示,TaKCS6基因编码的蛋白参与C24以上超长链脂肪酸的延伸。     

不同发育特性小麦品种春化基因 VRN2的序列分析

为了明确小麦春化基因VRN2与春化发育表现型的关系,以6个不同春化发育特性的普通小麦品种为试验材料,采用PCR技术对春化基因VRN2的CCT保守区中43个氨基酸序列进行了分析。结果表明,不同品种间ZCCT-A1的CCT功能域的序列存在差异,肥麦有R35W突变,另外5个品种均有R39C突变;ZCCT-A2均存在R16C突变;B和D基因组中均未发现突变。这表明VRN2基因编码区的等位变异主要出现在A基因组上,而B和D基因组中的VRN2基因在目前大面积主栽品种中均为显性。     

Phenolics and Antioxidative Activities in Narrow-Leafed Lupins (Lupinus angustifolius L.)

Eight lupin (Lupinus angustifolius L.) genotypes grown at four locations in south central Alberta in 2004 were evaluated for variability in phenolic constituents and antioxidant activity measured by a photochemiluminescence assay. Genotype was the main source of variation for content of phenolic compounds and antioxidant activities. Phenolic compounds in genotypes varied minimally from 11.9 to 14.7 mg catechin equivalent and 4.15 to 4.95 mg rutin equivalent g⁻¹ lupin for total phenolic and flavonoid contents, respectively. Lupin genotypes exhibited weak antioxidant activity based on water–soluble substances (ACW) of 0.54 to 1.07 μmole Trolox equivalent antioxidant capacities (TEAC)/g with lag time ranging from 70 to 153 s and an antioxidant index of 6.7 to 14.5 and 1.9 to 3.3 μmole TEAC/g based on measurements of lipid-soluble substances (ACL). Antioxidant activity of lupin genotypes was not related to phenolic contents of seeds.     

普通小麦TaTPR1基因的克隆及表达分析

具有TPR基序的蛋白质被认为能够介导蛋白质间的相互作用,并且参与多种生物学过程,如细胞周期调控、转录调控、过氧化物酶等蛋白质的运输、信号传导和蛋白质折叠等。为了在小麦分子育种研究中获得更多有价值的候选基因,本研究采用电子克隆和RT-PCR方法从小麦中克隆得到1个TPR类基因,命名为TaTPR1。该基因ORF长度972bp,推测编码包含323个氨基酸残基的蛋白,相对分子质量34.9kD,理论等电点为5.89。氨基酸序列分析表明,该蛋白在142~207区和207~274区分别含有TPR类基因家族特有的保守结构域(TPR_16和TPR_11)。进化和聚类分析表明,小麦TaTPR1基因与粗山羊草AtTPR15基因、乌拉尔图小麦TuTPR15基因的亲缘关系较近,蛋白相似度分别为91.28%和91.55%。Real-time PCR表达特性分析显示,该基因为组成型表达,在根、茎、叶中均表达;幼苗期茎中表达量较高,随幼苗的生长,茎中表达量上调显著;该基因表达受高盐的强烈诱导,也受水分、低温和外源ABA胁迫诱导。