The effect of different extraction techniques on extraction yield, total phenolic, and anti-radical capacity of extracts from Pinus radiata Bark

The performance of four techniques, conventional maceration, Soxhlet extraction, microwave assisted extraction (MAE), and ultrasound assisted extraction (UAE), for extraction of Pinus radiata bark, in one and several stages, were evaluated. For each technique, the mass extracted (g extract/g bark), total phenols (by Folin-Cicalteau), and tannin (by precipitation) concentration and anti-radical capacity (diphenyl-picrylhydrazyl, DPPH) were quantified. In one stage, the extracted mass increased in the following order: maceration < UAE < MAE < Soxhlet (p < 0.05). The total phenols and tannin levels were also higher with the Soxhlet technique. With additional extraction stages, only the samples produced with MAE and UAE techniques improved their parameters. Additionally, MAE extracts presented a higher anti-radical capacity than does Soxhlet and Pycnogenol^® extracts. Therefore, MAE was a simple and rapid method that was useful for extraction of P. radiata bark. Scanning electron micrographs (SEM) provided evidence of the mechanical effects on cell walls, mainly evidenced by cell destruction produced by Soxhlet, MAE, and UAE on the bark. In contrast, maceration only results in slightly ruptured cell pores, which could explain its low extraction yield.     

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.     

Effect of nixtamalization process on the content and composition of phenolic compounds and antioxidant activity of two sorghums varieties

Sorghum, a competitive crop due to its agronomical advantages, has gained interest as functional food. Sorghum contains phytochemicals with health benefits and presents some antinutritional compounds such as condensed tannins (CTs). However, it is well known that CTs exhibit antioxidant capacity. This study evaluated the effect of Mexican nixtamalization on total phenols, CTs and antioxidant capacity using two varieties of sorghum. Different conditions of lime (0, 1 and 2%) and cooking time (20, 30 and 40 min) were tested to obtain the best treatment after optimization using multiple linear regression analysis, aiming to minimize the CTs content and maximize the antioxidant capacity. CTs were reduced at 27% in white sorghum and 90%, in red sorghum. Total phenols, flavonoids and antioxidant capacity had an inverse correlation with lime concentration and cooking time. The best conditions for preserving antioxidant capacity with low content of tannins were 1.13% of lime and 31.11 min cooking. The phenolic profile obtained by HPLC showed reductions in gallic and chlorogenic acids only in white sorghum, whereas catechin and quercetin decreased in both varieties. We concluded that nixtamalization effectively reduced CTs to safety intake values, allowing to preserve other phenolic compounds and antioxidant capacity.     

Effect of growth stage on the content and composition of the essential oil and phenolic fraction of sweet marjoram (Origanum majorana L.)

The variation in the content and composition of Origanum majorana L. essential oil has been studied. Plant material has been harvested at four phenological stages (early vegetative, late vegetative, budding and full-flowering). Essential oil yield varied from 0.04 to 0.09% reached during the full-flowering stage. Analysis of the essential oils by GC and GC/MS revealed the presence of 38 components represented mainly by oxygenated monoterpenes (64.01–71.4%), monoterpene hydrocarbons (21.73–29.92%) and sesquiterpene hydrocarbons (1.47–4.05%). The main components were terpinen-4-ol (29.13–32.57%), cis-sabinene hydrate (19.9–29.27%), trans-sabinene hydrate (3.5–11.61%), γ-terpinene (2.11–8.20%), bornyl acetate (1.52–2.94%) and linalool (1.05–1.39%). On the other hand, phenolic contents varied from 2.706 to 6.834 mg/g of dry weight obtained during the later vegetative stage. RP-HPLC analysis of the methanolic extract of O. majorana L. dried aerial parts showed the predominance of phenolic acids during the early vegetative stage whereas flavonoids predominate during the other stages of growth. The main phenolic acids identified were trans-2-hydroxycinnamic, rosmarinic, vanillic, chlorogenic, gallic and cinnamic whereas the main flavonoids were amentoflavone, apigenin, quercetin, luteolin, coumarin and rutin. Results obtained showed that later vegetative stage is characterized by the highest contents of bioactive compounds and therefore it could be considered as the best stage for harvesting marjoram plants.     

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.     

Relationship Among Antimutagenic, Antioxidant and Enzymatic Activities of Methanolic Extract from Common Beans (Phaseolus vulgaris L)

Common beans are rich in phenolic compounds, which can provide health benefits to the consumer. The objective of this work was to study the relationship among antimutagenicity, antioxidant and enzymatic activities of methanolic extract and trolox by principal components multivariate analysis. Antimutagenicity of phenolic compounds present in methanolic extract from the seed coat of common beans (P. vulgaris, Flor de Mayo Bajío cultivar) and trolox against AFB₁ mutagenicity were evaluated in the Salmonella typhimurium microsuspension assay. Antioxidant capacity of methanolic extract and trolox were evaluated using β-carotene and 1,1-diphenyl-2-picryhydrazyl (DPPH) in vitro model assays. Cythrome P450 activity was measured by fluorometric assay. For phenolic extract, trolox and phenolic extract + trolox, the inhibition on AFB₁ mutagenicity in tester strain TA100 was 47, 59 and 69%, respectively. While in TA98 was 39, 48 and 68%. The inhibition of phenolic compounds, trolox and phenolic compounds + trolox on cytochrome P450 (CYP450) activity was 48, 59 and 88%, respectively. Correlation analysis showed that phenolic extract and trolox have high antimutagenic and antioxidant activity and also inhibited enzymatic activity. The results suggest that the primary mechanism of action of phenolic compounds in beans against AFB₁ mutagenicity may be extra-cellular in the microsuspension assay.     

The oil content and fatty acid composition of various genotypes of cauliflower,turnip and radish

Seeds of thirteen genotypes of cauliflower, four of turnip and seven of radish were analysed for oil content and fatty acid composition. Turnip and radish had higher oil content as compared to cauliflower. Oleic, linoleic + eicosenoic and erucic acids were the major fatty acids in the oils of these seeds. Levels of erucic acid were in the order of radish < turnip < cauliflower. Compared to cauliflower and turnip seeds, radish seeds contained higher amounts of palmitic, oleic, and linolenic + eicosenoic acids and lower amounts of linoleic and erucic acids.     

Salt effect on yield and composition of shoot essential oil and trichome morphology and density on leaves of Mentha pulegium

The aim of the present work was to study salt effect on the yield and composition of shoot essential oil (EO) and the structures responsible for its biosynthesis in Mentha pulegium L. Shoot EO was extracted by hydrodistillation and composition was determined by GC–MS method. Apical and basal leaves were taken for microscopy analyses; small fresh samples were observed directly without fixation or metallisation with environmental scanning electron microscope (ESEM) and stereomicroscope (SM). Fresh separate epidermis was used for light microscopy (LM). Salt stress enhanced EO yield by about 2.75 times and affected the percentage of menthone, which is the major compound (51%), increasing that of pulegone. Menthone, pulegone, and neomenthol constituting the monoterpene class were found to be the principal components. The anatomical study showed three types of trichomes: (i) non-glandular, multicellular, simple hairs; (ii) small, capitate glandular trichomes; (iii) and peltate glandular trichomes. In control plants, the density and size of trichomes varied with leaf side (abaxial or adaxial) and developmental stage. Salt stress results in significant modifications affecting trichome distribution and size on both sides.     

青稞法尼基转移酶β亚基编码基因HbERA1的克隆及表达分析

ERA1(Enhanced response to ABA)基因编码法尼基转移酶(Farnesyl transferase)β亚基,该酶在干旱胁迫下对ABA信号负向调控因子的修饰起着关键作用。本研究以青稞(Hordeum vulgare subsp.vulgare)抗旱品种喜马拉雅10号为材料,利用RT-PCR技术克隆获得了ERA1基因全长cDNA序列,命名为HbERA1(登录号:KJ699392)。生物信息学分析表明,该基因全长1 401bp,可编码466个氨基酸序列,蛋白分子量为51.14kD,等电点为5.00。Prosite Scan分析结果表明,HbERA1含有多个干旱胁迫响应蛋白的作用位点,如酪蛋白激酶Ⅱ磷酸化位点、N-糖基化位点、蛋白激酶C磷酸化位点及N-豆蔻酰化位点。利用实时定量PCR方法研究了HbERA1在干旱胁迫条件下及复水后不同时间点的表达情况,发现在水分过剩处理下(土壤绝对含水量15.5%),HbERA1在土壤绝对含水量为33.4%时表达量最高,并随着土壤绝对含水量的下降而下调表达;进行干旱胁迫后(15.5%)基因表达量也明显下调表达;复水后表达逐渐恢复,复水8h时超过正常表达水平,表明HbERA1基因可能参与调控水涝和干旱胁迫双重信号传导。     

玉米圆斑病拮抗菌芽孢杆菌21的鉴定及抑菌研究

利用形态特征、培养性状、生理生化特征及16Sr DNA序列分析,对获得的具有生防潜力的芽孢杆菌21进行鉴定,确定该菌株为枯草芽孢杆菌(Bacillus subtilis spp.)。利用显微观察法,研究枯草芽孢杆菌21对玉米圆斑病菌的抑制作用,结果显示,该菌株不同浓度发酵液处理后玉米圆斑病菌孢子萌发率出现下降趋势,萌发率最低为0;经发酵液处理后的玉米圆斑病菌芽管产生畸形,在发酵液浓度为90%时,芽管畸形最严重;处理后的玉米圆斑病菌菌丝出现畸形,菌丝中的内容物有外渗趋势。     

球孢白僵菌对亚洲玉米螟毒力生测方法比较研究

在球孢白僵菌不同浓度下,利用饲喂法和浸渍法测试球孢白僵菌对亚洲玉米螟3龄幼虫的致病力,建立球孢白僵菌对亚洲玉米螟毒力生测标准。结果表明,处理组中白僵菌浓度为10⁵、10⁶、10⁷、10⁸ CFU/mL时,采用饲喂法和浸渍法对3龄幼虫的校正死亡率、致死中时间（LT₅₀）、致死中浓度（LC₅₀）均没有显著性差异;饲喂法3龄幼虫的死亡率显著低于浸渍法。因此,球孢白僵菌对亚洲玉米螟毒力生测的常用方法可用饲喂法替代浸渍法,饲喂法试验过程简单,避免反复移动试虫引起的误差,毒力测试结果更可靠。     

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。     

Studies on life cycle of mealybug, Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae), on different hosts at different constant temperatures

The life cycle of a laboratory reared parthenogenic line of mealybug, Maconellicoccus hirsutus (Green) (Hemiptera: Pseudococcidae) was investigated on six host plants (Lagenaria siceraria var.clavata, Solanum tuberosum L. var. Kufri Chandramukhi, Praecitrullus fistulosus (Stocks) Pangalo var. MTNH1, Cucurbita pepo var. Styriaca, Triticum aestivum L. var. Lokvan, H. rosa sinensis L.) at five constant temperatures (25, 29, 35, 38, 42 °C). The number of emerging crawlers and adults as well as weight of adults varied with host and temperature. P. fistulosus and H. rosa sinensis were suitable hosts for laboratory rearing of M. hirsutus. However, ease of maintenance of P. fistulosus to produce M. hirsutus under laboratory conditions is an additional advantage compared to using green plants such as H. rosa sinensis. Among the range of constant temperatures, 38 °C was found to be the most favourable for the development and survival of M. hirsutus. These results suggest that P. fistulosus fruits are suitable hosts for laboratory rearing and life cycle studies of M. hirsutus.     

小麦 TaWIN1基因的克隆和表达分析

14-3-3蛋白家族在植物生长发育和逆境胁迫响应中发挥着重要作用。 TaWIN1基因是小麦14-3-3基因家族成员之一,为了进一步了解该基因的功能,本研究以普通小麦中国春为材料克隆了 TaWIN1基因并进行了生物信息学分析和表达分析。结果表明, TaWIN1基因含有801 bp的开放阅读框,编码266个氨基酸,含有完整的14-3-3蛋白家族结构域;该基因的编码蛋白为酸性蛋白,不具有跨膜区,可能定位于细胞质;进化分析显示,小麦TaWIN1蛋白与大麦14-3-3E、二穗短柄草GF14-D的亲缘关系最近; TaWIN1基因在小麦不同发育时期和不同组织中均有表达,但在10 mm幼穗中的相对表达量最高,其次为苗期叶片和旗叶,在根中表达量最低;根中 TaWIN1基因在干旱、高温、低温和盐胁迫下均显著上调表达;叶片中 TaWIN1基因在干旱、低温和盐胁迫下均显著上调表达,而在高温下则显著下调表达。     

Chemical composition and anti-fungal properties of the essential oils and crude extracts of Orthosiphon stamineus Benth

The hydrodistilled essential leaves and stems oils of Orthosiphon stamineus Benth were analysed by GC–MS/MS. Sixty nine compounds representing 97.6 and 97.4% of the total leaves and stems oils, respectively were identified, of which β-caryophyllene (24.0 and 35.1%), α-humulene (14.2 and 18.4%), β-elemene (11.1 and 8.5%), 1-octen-3-ol (8.2 and 7.0%), β-bourbonene (3.4 and 3.0%), β-pinene (2.1 and 1.7%), caryophyllene oxide (1.6 and 2.2%), camphene (1.6 and 1.3%) and limonene (1.2 and 1.1%) were the major compounds. Thus, the monoterpenes and sesquiterpenes were the predominant portions of the oils. Essential oils and methanol extract of O. stamineus and the derived fractions of hexane, chloroform, and ethyl acetate were tested for anti-fungal activity, which was determined by disc diffusion and minimum inhibitory concentration (MIC) determination methods. The oils, methanol extract and derived fractions of methanol extract displayed great potential of anti-fungal activity as a mycelial growth inhibitor against the tested phytopathogenic fungi such as Botrytis cinerea, Rhizoctonia solani, Fusarium solani, Colletotricum capsici and Phytophthora capsici, in the range of 49.3–70.3% and minimum inhibitory concentration ranging from 500 to 1000 μg/ml.     

Evaluation of corn plant as potential banker plant for supporting predatory gall midge, Feltiella acarisuga (Diptera: Cecidomyiidae) against Tetranychus urticae (Acari: Tetranychidae) in greenhouse vegetable production

The twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most important and highly polyphagous pests of vegetables and other crops worldwide. Experiments were conducted in the laboratory and greenhouse to evaluate corn (Zea mays L.) as a banker plant for the predatory gall midge, Feltiella acarisuga (Vallot) (Diptera: Cecidomyiidae) to potentially control T. urticae. Choice and no-choice experiments were carried out to determine the host plant preference of an alternative prey, Oligonychus pratensis (Banks)(Acari: Tetranychidae) to corn and green bean (Phaseolus vulgaris L.). Results showed that O. pratensis adults strongly preferred corn as a host plant and posed no risk to green bean. F. acarisuga was found to fly at least 7.0 m to search for new preys on green bean plants, and over 176 F. acarisuga larvae per leaf were recorded at 14 d after dispersal. F. acarisuga proved to be an excellent predator of both T. urticae and O. pratensis. The predation by F. acarisuga to T. urticae and O. pratensis ranged from 43.7 to 67.9% and 59.2 to 90.3%, respectively, under laboratory conditions. In a non-cage study, 81.2% of T. urticae population was suppressed by F. acarisuga in reference to the control (cage treatment). The results showed that this banker plant system has potential for controlling T. urticae in greenhouse vegetable production.     

Oil content and fatty acid composition of promising IndianBrassica campestris L. (Toria) genotypes

Seeds of sixty four genotypes ofBrassica campestris L. (Toria) were analyzed for oil content and fatty acid composition. Oil content varied from 38.9% to 44.6%. Major fatty acids viz. oleic, linoleic, linolenic, eicosenoic and erucic acids exhibited ranges from 10.1% to 17.3%, 5.9% to 14.5%, 5.2% to 15.0%, 7.7% to 13.7% and 39.6% to 59.9%, respectively. Compared to the standard cultivar ITSA, four genotypes contained 5 to 8% lower content of erucic acid; and six genotypes contained significantly higher content of linoleic acid. Oil was positively correlated with erucic acid. The observed inverse relationship between linoleic and erucic acid contents might be utilized in breeding nutritionally better Toria cultivars with both high linoleic and low erucic acid contents.     

Antioxidant and trace element content of damiana (Turnera diffusa Willd) under wild and cultivated conditions in semi-arid zones

Damiana (Turnera diffusa Willd) is traditionally used in the elaboration of tea, liquors, in the production of cosmetic products, and as a medicinal plant. The aim of this study was to evaluate the antioxidant content and trace element concentrations in leaves and stems from damiana grown wild and under cultivated conditions in semi-arid zones. Plants were collected at three different sites, two where damiana grows wild and one where it is cultivated for experimental purposes. Significant differences in the concentration of trace elements, vitamin C and total phenolic compound content, as well as antioxidant enzyme activities, total antioxidant capacity, and lipid peroxidation levels were found between leaves and stems, as well as among sampling sites. Environmental conditions, in combination with genetic factors, apparently affect the trace element concentrations, antioxidant content, and total antioxidant capacity in damiana grown wild. Results suggest that nutrient enrichment, via the addition of fertilizers, contributed to increased antioxidant defenses in the cultivated damiana plants. Furthermore, we suggest that genetic selection, controlled environmental conditions, and specific agrotechnical practices can increase the antioxidant yield in leaves and stems of cultivated damiana.     

小麦TaHTAS-5A基因的克隆、表达分析及亚细胞定位

非生物胁迫对小麦的生长发育具有不良影响,克隆与非生物胁迫相关的基因,并对其结构及表达特性进行分析,可以为进一步探索小麦的抗逆机制和抗逆育种奠定基础。本研究利用同源克隆技术从普通小麦品种中国春中克隆了水稻E3泛素连接酶基因OsHTAS的同源基因TaHTAS-5A(GeneBank登录号:MF967573),并利用生物信息学方法对其基因序列特征进行分析;利用qRT-PCR技术对该基因在小麦不同组织及不同逆境胁迫下的表达模式进行分析;同时,对该基因的表达产物进行了亚细胞定位。结果表明,小麦TaHTAS-5A基因含有220bp的5′UTR、1 242bp的ORF以及126bp的3′UTR,共编码413个氨基酸;基因组分析表明,该基因全长3 819bp,共包含4个外显子和3个内含子;缺体-四体定位表明该基因位于小麦5A染色体上;蛋白结构分析显示,TaHTAS-5A蛋白的N端包含四个跨膜结构域,C端包含一个E3泛素连接酶特有的RING finger保守结构域,具有植物E3泛素连接酶的结构特征;qRT-PCR结果显示,TaHTAS-5A对高温、低温和ABA都有响应,对干旱和盐响应不敏感;TaHTAS-5A在小麦的根、茎、叶和幼穗等不同组织中均有表达,但表达量有差异,在叶和穗中的表达量明显高于根部和茎部;亚细胞定位结果表明,TaHTAS-5A位于细胞膜上。本研究为进一步探究小麦TaHTAS-5A基因的功能奠定了基础,也为小麦非生物胁迫抗性改良提供了一定的理论依据。