In vivo antioxidant and hepatoprotective activity of methanolic extracts of Daucus carota seeds in experimental animals

Objective:To assess the In vivo anlioxid Fanl and hepaloproleclive activity of metlianolic exlracl of Daucus carota(D.carota) seeds in experimental animals.Methods:Methanolic extracts of D.carota seeds is used for hepatoproleclion assessment.Oxidative stress were induced in rats by thioacetamide 100 nig/kg s.c.in four groups of rats(two test,standard and toxic control). Two test groups received D.carota seeds extract[DCSE) at doses of 200 mg/kg and 400 mg/kg. Standard group received silymarin(25 mg/kg) and toxic control received only thioacetamide. Control group received only vehicle.On the 8th day animals were sacrificed and liver enzyme like serum glutamic pyruvic transaminase(SGPT),serum glutamic-oxaloacetic transaminase(SCOT) and alkaline phosphatase(ALP)were estimated in blood serum and antioxidant enzyme like superoxide disnuituse(SOD),cululase(CAT),glutathione reductase(CKD),glutathione peroxidase(GPX),glutalhione-S-transferase(GST)and lipid peroxidation(LPO)were estimated in liver homogcnatc.Results:A significant decrease in SGPT,SCOT and ALP levels was observed in all drug treated groups as compared to thioacetamide group(P<0.001) and in case of antioxidant enzyme a significant(P<0.001) increase in SOD.CAT,GRD,GPX and GST was observed in all dmg treated groups as compared with thioacetamide group.But in case of LPO a significant(P <0.001) reduction was observed as compared to toxic control group.Conclusions:DCSE has contributed lo the reduction of oxidative stress and the protection of liver in experimental rals.

Chemical analysis,antimicrobial and anti-oxidative properties of Daucus gracilis essential oil and its mechanism of action

Objective:To evaluate the essential oils(EO) composition,antimicrobial and antioxidant power of a local plant,Daucus gracilis(D.gracilis).Methods:The aerial parts of D.gracilis were subjected to hydro distillation by a Clevenger apparatus type to obtain the EO which had been analyzed by gas chromatography and gas chromatography coupled with mass spectrometry,and screened for antimicrobial activity against five bacteria and three fungi by agar diffusion method.The mechanism of action of the EO was determined on the susceptible strains by both of time kill assay and lysis experience.The minimal inhibitory concentrations were determined by agar macrodilution and micro-dilution methods.Anti-oxidative properties of the EO were also studied by free diphenyl-2-picrylhydrazyl radical scavenging and reducing power techniques.Results:The EO yielded 0.68(v/w).The chemical analysis presented two dominant constituents which were the elemicin(35.3%) and the geranyl acetate(26.8%).D.gracilis EO inhibited the growth of Bacillus cereus and Proteus mirabilis significantly with minimal inhibitory concentrations of 17.15 μg/mL by the agar dilution method and57.05 μg/mL and 114.1 μg/mL,respectively by liquid micro-dilution.A remarkable decrease in a survival rate as well as in the absorbance in 260 nm was recorded,which suggested that the cytoplasm membrane was one of the targets of the EO.The EO showed,also,important anti-oxidative effects with an IC50 of 0.002 mg/mL and a dosedependent reducing power.Conclusions:D.gracilis EO showed potent antimicrobial and anti-oxidative activities and had acted on the cytoplasm membrane.These activities could be exploited in the food industry for food preservation.

Early Identification of Stable Transformation Events by Combined Use of Antibiotic Selection and Vital Detection of Green Fluorescent Protein (GFP) in Carrot (Daucus carota L.) Callus

Genetic transformation is a useful technique to complement conventional breeding in crop improvement. Although carrot has been a model organism for in vitro embryogenesis study, genetic transformation of carrot is still lengthy and labor intensive. An efficient transformation and detection system is desirable. Direct infection of Agrobacterium to carrot calli has provided an easy way for carrot genetic transformation. To improve the efficiency of antibiotic selection in this method, we report the combined use of an improved green-fluorescent protein, referred to as smGFP, to establish a versatile selection method for carrot callus transformation system. By combining antibiotic selection with the bright fluorescence observed in the callus tissue, we were able to easily identify stable transformants in early stage of the transformation process. In addition to the GFP expression of the callus cells, the transgenic nature of callus cells was confirmed with Southern and Western analysis. We found we can link the simplicity of carrot-callus-cell transformation, early detection of stable transformants with antibiotic selection, visualization of GFP fluorescence, and molecular analysis （Southern and Western） of callus tissue （non-photosynthetic tissue） to provide a more efficient way in identifying stable transformants at early stage of carrot transformation.

Components of Essential Oils in Different Parts of Daucus carota L. var. sativa Hoffm

The components of the essential oils obtained from different parts of Daacus carota L.var. saliva Hoffm were analyzed. The percentages of the essential oils extracted are 0. 27% （ mL/100 g material） for the flowers, 0. 0/% for the stems and leaves and 0. 01% for the roots. Fifty-four, Sixty-six and Thirty-three compounds were,respectively, separated and identified from the flowers, the stems and leaves and the roots, among which unsaturated alkene compounds are thirty-nine, thirty-nine and twenty-one, respectively, accounting in turn for up to 90. 21%, 90. 49% and 72. 65% of the total essential oils. Because alkene compounds have double bonds that are easily oxidized, it can he inferred that the components of the essential oils in the different parts of Daucus carota L. var. saliva Hoffm should show an activity of the antioformation of free radicals to some extent.

Asymmetric Reduction of Heteroaryl Methyl Ketones Using <i>Daucus carota</i>

Asymmetric reduction of the heteroaryl prochiral ketones to corresponding chiral alcohols by Daucus carota was studied. The study highlights selective bioreduction of different substituted heteroaryl ketones (1a - 1j) to their respective chiral alcohols (2a - 2j) using plant dehydrogenase enzymes present in Daucus carota in good yields (60% - 95%) and enantioselectivity (76% - 99%) with S-form configuration. The results obtained confirm that the membrane bound dehydrogenase enzyme has broad substrate specificity and selectivity in catalyzing both six and five membered heteroaryl methyl ketones. The present methodology demonstrates promising and alternative green route in the synthesis secondary chiral alcohols of biologically importance in a simple, inexpensive and eco-friendly process.

胡萝卜(Daucus carota var. sativa)密码子偏好性及属内质体基因组核苷酸多样性分析

核苷酸多样性作为衡量物种遗传变异水平的重要指标,对于研究物种遗传多态性的高低、进化历史和系统位置具有重要的科学意义,密码子偏好性则跟基因表达和自然选择压力等密切相关。本研究通过计算胡萝卜属(Daucus L.) 17种植物的叶绿体基因组片段的核苷酸多样性,开发出了可用于系统学和群体遗传学等研究的叶绿体基因分子标记;并且基于Codon W和CUSP统计数据,通过相关性分析、中性绘图分析、ENC-plot分析以及PR2-plot分析,探究了胡萝卜(Daucus carota var. sativa)密码子使用偏好性。结果表明,有效密码子数(ENC)与最优密码子使用频率(FOP)、密码子适应指数(CAI)、密码子偏好指数(CBI)具有显著的相关性,胡萝卜叶绿体基因的密码子偏好性主要受到突变和自然选择的作用,其中,突变的影响更大。此外,本研究确定了5个高表达基因和18个最优密码子。这些研究结果有助于探究胡萝卜的进化历史和基因的表达,从而为品种改良和提高产量提供理论支持。

Isolation of development-related genes in somatic embryo radicle of carrot (Daucus carota L.) using modified cDNA RDA

Using modified cDNA RDA capitalizing on the high affinity of streptavidin for biotin and magnetic-absorption-based separation, we have obtained four bands of specifically expressed cDNA in the carrot somatic embryo deregulated for 12 h, which were designated as NR-1, NR-2, NR-3 and NR-4, respectively. As revealed by homology analysis of their DNA sequences after cloning them into pBS, remarkable homology was demonstrated in NR-2, NR-3 and NR-4 with the genes coding for LEA (late embryogenesis abundant protein), Dna J and xyloglucan endo-trans-glycosylase in plants. On the other hand, NR-1 showing no homology with any known sequence may have come from unknown genes. Using 32P-labeled NR-1 as probe, hybridization with cDNA fragment population has shown that we have actually cloned a new gene fragment related to radicle development. As shown by further Southern hybridization, these genes may be present in carrot genome in the form of single or low copies.

Dc GA20ox2 and DcGA2ox1 alter endogenous gibberellin contents and adjust lignin accumulation in carrot

Gibberellins(GAs)are a class of plant hormones that can affect plant growth and development.GA-oxidases are rate-limiting enzymes,which play a direct role in GA accumulation in plants.However,the roles of GA-oxidase on carrot(Daucus carota L.)taproot development are still unclear.In this study,two GA-oxidase genes,DcGA20ox2 and DcGA2ox1,were identified in carrot.Transgenic carrot plants were obtained by using Agrobacterium-mediated genetic transformation method.The results showed that overexpression of DcGA20ox2 significantly promoted the accumulation of active GAs in carrot,increased plant height,generated more branches,and enhanced xylem development.Overexpression of DcGA2ox1 significantly reduced the total contents of active GAs compared with the control group,resulting in a dwarf phenotype and markedly increased lignin content of the transgenic carrot.The expression profiling showed that the genes of GA metabolic pathway responded to the negative feedback regulation mechanism.At the same time,the expression of most genes in lignin biosynthesis and polymerization process was up-regulated,corresponding to the massive accumulation of lignin.These findings indicated that DcGA20ox2 and DcGA2ox1affected carrot growth and development by regulating the levels of endogenous GAs.The results from current work might shed light on further studies aimed to regulate lignification in carrot and other crops.

Synthesis of betanin by expression of the core betalain biosynthetic pathway in carrot

Betalain has received increased attention because of its high nutritional value and crucial physiological functions.Based on the elucidation of its core biosynthetic pathway,betalain can be produced in additional plants by metabolic engineering.Synthesis of betalain in carrot(Daucus carota L.)can improve its nutritional quality and economic value by extracting betalain from the fleshy root,non-edible part,and processing residue of carrot.In this study,two different constructs,namely,pYB:mCD(AomelOS,BvCYP76AD1S,and BvDODA1S)and p YB:CDD(BvCYP76AD1S,BvDODA1S,and MjcDOPA5GTS),were introduced into carrot for betanin synthesis by Agrobacterium-mediated transformation.Betanin can be synthetized in both transgenic calli,and p YB:m CD-transgenic callus can be used to produce betacyanin by suspension culture.However,pYB:mCD-transgenic seedlings can synthetize betanin only by tyrosine feeding.The p YB:CDD-transgenic lines can synthetize betanin in whole plants.The betanin content in fleshy root of pYB:CDD-transgenic carrot was(63.4±9)μg·g^(-1)fresh weight according to quantitative analysis.These betanin-producing carrot plant materials can be used to synthesize betanin for industrial application or consumption as dietary sources.

胡萝卜抗冻蛋白在大肠杆菌中的高效表达

以pET - 11a为载体构建了胡萝卜抗冻蛋白非融合表达的重组质粒 ,并在大肠杆菌菌株BL2 1(DE3,pLysS)中成功地进行了高效表达。表达的蛋白质以包涵体的形式存在 ,相对分子质量为 36 80 0。经正交试验得到了优化的诱导表达条件。包涵体经各种缓冲液洗涤纯化、溶解、复性和超滤浓缩后 ,经SDS -PAGE电泳检测 ,纯度达单一带水平。生物学活性研究表明 ,重组表达的胡萝卜抗冻蛋白具有很强的热滞活性和提高细菌耐寒性的作用。

不同包装处理对胡萝卜贮藏期间品质的影响

研究不同包装方式下的胡萝卜(Daucus carota)在贮藏期间的品质变化,筛选出最佳的包装方式以有效延长胡萝卜的贮藏时间。结果表明,塑料箱加微孔膜与塑料箱加0.04 mm薄膜包装可有效减缓胡萝卜的失重率、糠心指数、颜色指数、游离氨基酸质量分数、粗纤维质量分数、总酚物质质量分数的升高,减缓可溶性蛋白质质量分数的降低,适合应用于胡萝卜的贮藏;塑料箱、纸箱、编织袋包装的胡萝卜品质下降明显,不适合用于胡萝卜的长期贮藏。

响应面优化超声辅助提取胡萝卜叶多酚工艺研究

为提高胡萝卜(Daucus carota)叶的利用率,对胡萝卜叶多酚的提取工艺进行了研究。以多酚提取率为响应值,以超声时间、乙醇浓度、液料比和超声温度为因素,利用响应面法对实验进行设计及优化。结果表明,胡萝卜叶多酚的最佳提取工艺为:超声时间41min、乙醇浓度71%、液料比37mL/g和超声温度66℃。在此条件下,多酚提取率理论值为31.79mg/g,实际测得多酚提取率为32.13mg/g,与理论值相对误差为1.07%。

胡萝卜4CL基因家族的鉴定及分析

旨在全基因组水平上鉴定胡萝卜4CL基因,揭示该基因家族成员的理化性质、系统进化、基因结构、组织表达模式及胁迫响应情况,为胡萝卜4CL的功能研究和利用奠定基础。基于AMP结合结构域(PF00501)的隐马尔可夫模型文件,利用HMMER软件对胡萝卜蛋白质数据库进行检索,获得胡萝卜4CL基因(命名为Dc4CL);利用ProtParam分析Dc4CL的理化性质;利用MEGA6对Dc4CL进行系统进化分析;使用GSDS2.0、MEME分析Dc4CL的基因及蛋白结构;利用荧光定量PCR技术对Dc4CL的组织表达模式及胁迫响应情况进行研究。结果表明,胡萝卜基因组中存在12个4CL基因,分别为Dc4CL1~Dc4CL12。Dc4CL基因开放阅读框(ORF)长度为1350~3363 bp,编码蛋白质长度为449~1120 aa,编码蛋白的分子质量和等电点分别为48.92~123.73 ku和5.34~8.82。系统进化分析表明,Dc4CL与其他来自水稻、拟南芥、二穗短柄草、高粱、大白菜的4CL蛋白可分为两个类群。结构分析表明,Dc4CL基因外显子数目为4~11个,其蛋白序列中均含有BoxⅠ(SSGTTGLPKGV)和BoxⅡ(GEICIRG)两个保守序列。荧光定量PCR结果表明,Dc4CL3、Dc4CL12在胡萝卜叶片及肉质根中均有表达,而Dc4CL8主要在叶片中表达,盐胁迫和低温胁迫均使Dc4CL3、Dc4CL8、Dc4CL12的表达降低。总之,胡萝卜的12个Dc4CL基因在影响4CL蛋白功能的关键位点处具有保守性,而在其他位置上存在一定的差异,且部分成员在逆境胁迫应答过程中可能发挥了一定的作用。

胡萝卜DcZDS基因的克隆和表达分析

以黄色胡萝卜‘黄珊瑚’(19071)、橙色胡萝卜‘新黑田五寸’(C1)、红色胡萝卜‘玫瑰红’(17106)为试材,采用生物信息学方法,研究了3个品种胡萝卜克隆得到的DcZDS基因编码的氨基酸组成和理化性质及其肉质根在不同发育阶段DcZDS基因的相对表达量,以期为DcZDS基因对番茄红素合成的影响提供参考依据。结果表明:3个品种胡萝卜DcZDS基因的cDNA全长均为1722 bp,编码573个氨基酸;DcZDS基因编码蛋白的理化性质基本相同,均属于亲水性且不稳定的蛋白;编码蛋白的二级结构主要包含α-螺旋和无规则卷曲,且主要在叶绿体中发挥作用,是一种非分泌性蛋白;系统进化树分析发现,胡萝卜DcZDS基因与芹菜DcZDS的亲缘关系最近。在肉质根发育的不同时期DcZDS基因的表达量均呈现先升高后降低的模式,但在肉质根发育的65 d时,DcZDS在红色肉质根中的表达量显著高于橙色和黄色肉质根,说明DcZDS的高表达促进了番茄红素在红色胡萝卜中的积累。

Nutritional and Health Benefits of Carrots and Their Seed Extracts

Carrot is a root vegetable with carotenoids, flavonoids, polyacetylenes, vitamins, and minerals, all of which possess numerous nutritional and health benefits. Besides lending truth to the old adage that carrots are good for eyes, carotenoids, polyphenols and vitamins present in carrot act as antioxidants, anticarcinogens, and immunoenhancers. Anti-diabetic, cholesterol and cardiovascular disease lowering, anti-hypertensive, hepatoprotective, renoprotective, and wound healing benefits of carrot have also been reported. The cardio- and hepatoprotective, anti-bacterial, anti-fungal, anti-inflammatory, and analgesic effects of carrot seed extracts are also noteworthy. All are discussed in this review article.

植物细胞的冷藏

日本科学家 N.Kaimori 和 N.Takahashi 披露了通过室温干燥。在低温保藏前从胡萝卜(Daucus carota)除去易结冰水分的效应.于27℃将置于含硅胶平皿中的胡萝卜下胚轴愈伤组织干燥至含水浓度3%.

A Simple Inexpensive Method for the Measurement of Indoleacetamide Hydrolase Activity

Indole-3-acetic acid (IAA) a phytohormon of auxin type is synthesized by different ways in plants and some bacteria (Agrobacteria, Pseudomonas and some others). The enzyme indolylacetamide hydrolase transforms indolylacetamide to IAA mainly in bacteria. However, recently published data showed that some plants can also hydrolyze indolylacetamide into IAA. In order to elucidate the role of indolylacetamide as an auxin precursor in plants and bacteria, productive method of determination of the activity of indoleacetamide hydrolase is necessary. The simple, inexpensive and productive method for the measurement of indoleacetamide hydrolase activity was elaborated based on significant difference between IAAM and IAA in color developed with Salkovski’ reagent. The light absorbance increased during conversion of IAAM to IAA by protein extracts from some plant cells and this increase may be used for quantitative estimation of indoleacetamide hydrolase activity. The method is suitable for fast discovery of indoleacetamide hydrolase activity before planning more complicated analyses and for the analysis of many probes at a short time in physiological and biochemical experiments. A detailed protocol for determination of indoleacetamide hydrolase activity by the elaborated method is described.

阳台巧种胡萝卜

胡萝卜（Daucus carota）别名红根、黄萝卜。原产阿富汗,主要分布欧洲与亚洲,以俄罗斯和中国栽培面积最大。我国南北均可种植。伞形花科胡萝卜属,二年生草本植物。株高40~60厘米。直根系,直根肥大,入土深。叶片着生于短缩的茎上,为2~3回羽状复叶,小叶狭披针形,绿色,被茸毛,叶柄较长。

Age-stage, two-sex life tables of Bactrocera cucurbitae （Coquillett） （Diptera： Tephritidae） with a discussion on the problem of applying female age-specific life tables to insect populations

Age-stage, two-sex life tables of the melon fly, Bactrocera cucurbitae （Coquil- let-t） （Diptera： Tephritidae）, reared on cucumber （Cucumis sativus L.）, sponge gourd （Luffa cylindrica Roem） and a carrot medium （mashed Daucus carota L. mixed with sucrose and yeast hydrolysate） were constructed under laboratory conditions at 25 4- loc, 65% 4- 0.5% relative humidity, and a photoperiod 12 ： 12h （L ： D）. The intrinsic rates of increase of B. cucurbitae were 0.144 6, 0.141 2 and 0.068 8 days on cucumber, sponge gourd, and carrot medium, respectively. The highest net reproduction rate was 172 offspring per fly reared on sponge gourd. The mean generation times ofB. cucurbitae ranged from 34 days reared on cucumber to 56 days reared on carrot medium. The life history raw data was analyzed using the traditional female age-specific life table and compared to results obtained using the age-stage, two-sex life table. When the age-specific female life table is applied to an age-stage-structured two-sex population, survival and fecundity curves will be improperly manipulated due to an inability to include variation in preadult development time. We discussed different interpretations of the relationship between the net reproductive rate and the intrinsic rate of increase to clarify possible misunderstanding in the literature.

基于转录组序列的胡萝卜EST-SSR标记开发及遗传多样性分析

胡萝卜(Daucus carota var. sativa)是一类重要的全球性蔬菜作物。为了全面了解其转录组SSR位点特点,并开发胡萝卜EST-SSR分子标记,本试验对胡萝卜的根、茎、叶混合组织进行转录组测序分析,共获得42 668条Unigene,其中6 611条Unigene序列中含有8 241个SSR位点,SSR发生频率为15.49%,平均分布为0.665 0 kb。主要优势重复序列为二核苷酸,占总SSR的45.35%。以48份胡萝卜为材料,从60对SSR引物中筛选出21对条带清晰稳定、多态性高的引物,其中有11对引物多态信息含量(PIC)大于0.500 0,属于高多态性引物,这些引物占开发引物的52.38%,表明筛选出的21对SSR引物具有较好的遗传多样性。聚类分析可将48份胡萝卜材料在相似系数为0.687 2处分为四类,聚类结果与胡萝卜颜色和产地存在一定的相关性。试验结果为胡萝卜的种质资源鉴定、品种鉴别和分子标记辅助育种等研究提供理论依据。