普通菜豆(Phaseolus vulgaris L.)AOX基因家族鉴定与表达分析

交替氧化酶(Alternative oxidase,AOX)是一种末端氧化酶,在维持植物正常生长代谢速率和代谢平衡以及抵御逆境胁迫中发挥了一定作用。AOX家族成员已经在很多植物物种中被鉴定,但是在普通菜豆(Phaseolus vulgaris L.)中却未见报道。研究对普通菜豆中AOX家族成员进行了鉴定和分析,并进一步解析了AOX基因家族成员的功能及调控机制。结果表明,基于普通菜豆参考基因组中共挖掘得到了4个AOX家族成员,通过进化关系、保守基序(Motifs)、基因结构分析,将各成员划分成4个亚族。此外,顺式作用元件、共线性与盐胁迫下的表达量分析结果表明,AOX成员可能在植物的非生物胁迫响应中发挥了作用。为后续深入研究普通菜豆中AOX成员的功能奠定了理论基础。

Effect of Biostimulants Based on Natural Products on the Growth and Nutritional Value of Bean (Phaseolus vulgaris L.)

Beans (Phaseolus vulgaris L.) are widely grown in Cameroon and play a key role in the fight against food insecurity, malnutrition and poverty. However, its cultivation encounters problems due to abiotic and biotic stresses, which leads to the use of synthetic fertilizers and pesticides, which cause significant damage to the environment and human health due to the presence of synthetics residues in the seeds, pods and in the leaves that are eaten. Promoting the use of natural products is becoming a necessity for organic and eco-responsible agriculture that limits contamination problems and improves people’s purchasing power. This study aims to assess the effect of biostimulants based on natural products on the growth and nutritional value of common bean (Phaseolus vulgaris L.). Bean seedlings from white variety (MEX-142) and red variety (DOR-701) were treated every seven days in the field from their pre-emergence, emergence and growth to their maturation under a randomized block experimental design. Six treatments and three repetitions with the biostimulants based on natural products and controls were thus performed and the agromorphological parameters were measured. After 120 days, the contents of growth biomarkers and defense-related enzymes were evaluated in leaves, while the contents of macromolecules, minerals and antinutrients were evaluated in seeds. These biostimulants significantly increased (P P < 0.0001) of antinutrients including oxalates, phytates, tannins and saponins in seeds compared to controls (T+ and T−). Treatment with biostimulants, in particular BS4, improves the performance of bean plants in the field as well as the biofortification of seeds regardless of the variety.

欧洲千里光(Senecio vulgaris Linn.)对干旱环境的适应

采用石蜡切片法,研究了欧洲千里光(Senecio vulgaris Linn.)营养器官的显微结构,以探讨其对干旱环境的适应性特点。结果表明:根和茎木质部导管发达,提高了输导能力;薄壁组织丰富,提高了储水能力;叶片表皮细胞外切向壁角质层发达,有利于减少水分散失;同化组织发达,能够适应干旱环境对光合作用的负面影响。欧洲千里光长期生活在干旱环境中,产生了许多适应性结构。

Photosynthetic Gas Exchange and Nitrogen Assimilation in Green Bean Plants Supplied with Two Sources of Silicon

Beans contain a wide range of vitamins,proteins,calcium,and zinc which make them an important food source for many countries.To meet the demand for bean production worldwide,large amounts of fertilizers and pesticides are used.However,the cost of production and environmental impact increases.To produce food sustainably,the use of beneficial nutrients such as silicon as a biostimulant has been proposed.However,information about the effect of different sources of silicon on the metabolism of bean plants is scarce.Bean plants cv.Strike were grown in pots for 60 days and the effect of foliar application of silicon nanoparticles and the silicon-based biostimulant Codasilat 4 concentrations(0,1,2,and 4 mM)on total biomass,yield,photosynthetic pigment concentration,photosynthetic activity,stomatal conductance,transpiration rate,chlorophyll fluorescence,and nitrogen assimilation were evaluated.The results obtained showed that the supply of silicon at a dose of 1 mM functioned as a biostimulant,favoring gas exchange and nitrogen assimilation within the plant,which stimulated growth and yield.The results of this research work allowed a better comprehension of the effects of silicon application through silicon nanoparticles and the biostimulant Codasilon the physiology of green bean plants.

欧洲千里光SvAPETALA1-5基因对花器官发育的影响

菊科(Asteraceae)是真双子叶植物的一大类,具有复杂的头状花序,目前对其花发育的分子调控机制还知之甚少。APETALA1(AP1)属于MADS-box基因家族,在植物成花诱导和花器官发育中具有关键作用。本研究对欧洲千里光(Senecio vulgaris)AP1同源基因SvAP1-5进行了生物信息学及组织特异性表达分析,并基于VIGS技术创建了SvAP1-5基因沉默植株,并在龙葵(Solanum nigrum)中过表达SvAP1-5,对其功能进行了初步探讨。生物信息学分析结果表明,SvAP1-5基因具有典型的MADS-box和K-box结构,C末端含有FUL基序和paleoAP1基序,属于euFUL分支的AP1类基因。qRT-PCR结果表明,SvAP1-5在苞片、花瓣和心皮中均有表达,其中在心皮内较高表达。与对照相比,SvAP1-5基因沉默导致花瓣不规则裂开,柱头分裂增加,心皮发育不良。而SvAP1-5过表达龙葵植株的果实发育异常,出现类似果实细胞的增生组织。综上所述,在欧洲千里光花发育过程中,SvAP1-5基因可能参与花瓣和心皮的发育,并发挥着重要作用。

超富集植物叶用红菾菜(Beta vulgaris var.cicla L.)及其对Cd的富集特征

超富集植物是重金属污染土壤植物提取修复的基础,超富集植物的筛选一直都是污染土壤植物修复研究的热点及前沿。采用盆栽实验对7种农作物积累Cd的特征进行研究,结果表明,叶用红菾菜对Cd具有较高的富集系数和转移系数。在此基础上,采用盆栽浓度梯度试验对叶用红菾菜对Cd的富集潜力及富集特征进行了系统研究。结果表明,在Cd污染水平为20mg·kg^(-1)条件下,叶用红菾菜地上部Cd含量超过了100mg·kg^(-1)这一国际公认的Cd超积累植物应达到的临界含量标准;且其对Cd的富集系数和转移系数均大于1;同时,与对照相比,植物的生长未受到抑制,这表明叶用红菾菜是一种Cd超积累植物,为Cd污染土壤的植物修复提供了一种新的种质资源。

低温诱导甜菜(Beta vulgaris L.)Ty7Br600基因的Northern blotting和Southern blotting分析

以低温诱导甜菜幼苗为试验材料,以未进行低温诱导的甜菜幼苗为对照,采用α-32P-ATP放射标记按随机引物标记法标记本实验室克隆的Ty7Br600基因,进行Northern杂交。Northern杂交试验结果显示,在低温诱导培养的甜菜幼苗的RNA群体中,出现较强的杂交条带,而在未诱导培养的甜菜幼苗的RNA群体中,则几乎没有阳性杂交条带出现。因此,Ty7Br600这一序列有可能是二年生甜菜幼苗经低温诱导处理之后被诱导表达的与抽薹相关的新基因序列。Southern杂交结果发现,在每一组酶切中至少有2~3条条带,表明Ty7Br600确为甜菜基因组片段,也同时表明该基因在甜菜基因组中以2个拷贝或低拷贝形式存在。

Comparative Experiments on Different Varieties of Phaseolus vulgaris L. under Dry Farming Conditions

[Objective] The aim was to screen varieties of Phaseolus vulgaris L.for cultivation in arid,semi-arid areas.[Method] The cultivation experiments of seven varieties of P.vulgaris under dry farming conditions were carried out in typical dry farming agriculture region Yangqu County in North China.[Result] The plants of Teyoutejiadou,Chaojiwujing and Xinxinyichiqingyuanlong were relatively strong;the growth rate of Aifeng,Jingxuanjiadouwang and Chaochangsijidou was relatively rapid;the transpiration rate of functional leaves of Aifeng was the lowest.The yield of Tianma 95-33 jiadouwang was significant higher than that of other varieties,the unit yield was up to 36.33 t/hm2,which was 1.39 times higher than that of Aifeng and 1.9 times higher than that of Xinxinyichiqingyuanlong;followed by the yield of Aifeng（26.07 t/hm2）,which was 1.37 times higher than that of Xinxinyichiqingyuanlong.[Conclusion] The yield of Tianma 95-33 jiadouwang and Aifeng was relatively high,as well as the protein content,indicating that they were suitable to be applied and cultivated in the experimental plot and surrounding areas.

L-肉碱对普通小球藻Chlorella vulgaris种群增长和生化组成的影响

分别以0,5,50 mg/L的L-肉碱(L-carnitine)处理淡水普通小球藻(Chlorella vulgaris),以SE为培养液,培养7 d,探究L-肉碱对淡水普通小球藻种群增长、总脂和可溶性蛋白质含量的影响。结果表明:不同质量浓度的L-肉碱在不同培养时期对普通小球藻种群增长影响显著(P<0.05);高浓度L-肉碱显著降低总脂含量(P<0.05),并使可溶性蛋白质含量显著升高(P<0.05)。结果提示,在培养液中添加外源L-肉碱可能通过直接作用于普通小球藻,或促进培养液中微小生物分泌活性物质间接影响普通小球藻的种群增长及总脂和可溶性蛋白质的含量。

紫冠豆角(Phaseolus vulgaris L.)种子中α-淀粉酶抑制剂的提取及其性质研究

本研究采用盐溶的方法从紫冠豆角(Phaseolus vulgaris L.)种子中提取α-淀粉酶抑制剂(α-AI),将提取的α-AI用不同温度和pH处理,并评价其稳定性。在此基础上,以提取物对猪胰腺α-淀粉酶的抑制活性(IC_(50))为指标,分析了豆粉的细度(A)、料液比(B)、盐溶时间(C)三个因素对α-AI提取效果的影响,利用响应面分析法优化提取条件,通过对老豆角种子的深加工,实现提高其附加值的目的。结果表明:该α-AI为耐热性蛋白,在pH4~10的范围性质稳定,影响α-AI的IC_(50)因素按主次顺序排列为:豆粉的细度>料液比>盐溶时间,确定提取α-AI最佳工艺条件为:豆粉过60目筛(粒径<0.3 mm),料液质量体积比为1∶12(g/mL)、盐溶时间7.75 h。紫冠豆角种子α-AI的IC_(50)最优值为27.036±0.235μg/mL。

Gas chromatography-mass spectrometry analysis of different organic crude extracts from the local medicinal plant of Thymus vulgaris L

Objective:To isolate and analyze the chemical composition in different crude extracts of from the leaves of locally grown of Thymus vulgaris L(T.vulgaris)by gas chromatography-mass spectrometry(GC-MS).Methods:The shade dried leaves powder was extracted with methanol by using Soxhlet extractor.Methanol crude extracts of T.vulgaris and the derived fractions of hexane,chloroform,ethyl acetate and butanol were obtained.Results:Qualitative analyses of various organic crude extracts of T.vulgaris by using GC-MS showed that there were different types of high and low molecular weight compounds.Most of the isolated and identified compounds by GC-MS in the crude extracts are basically biologically important.Further,the T.vulgaris leaf possessed certain characteristics that can be ascribed to cultivation on a domestic plantation.The crude extracts were prepared from the powder leaves of T.vulgaris for respective compounds can be chosen on the basis of above GC-MS analysis.Conclusions:All the major compounds were identified and characterized by spectroscopic method in different organic crude extracts of T.vulgaris are biologically active molecules.Thus the identification of a good number of compounds in various crude extracts of T.vulgaris might have some ecological role.

Hypoglycaemic effect of Berberis vulgaris L.in normal and streptozotocin-induced diabetic rats

Objective:To achieve a primary pharmacological screening contained in the aqueous extract of Berberis vulgaris(B.vulgaris) and to examine the hypoglycaemic effect and biochemical parameters of aqueous and saponins extract on groups of rats rendered diabetic by injection of streptozotocin.Methods:The phytochemicol tests to detect the presence of different compounds were based on the visual observation of color change or formation of precipitate after the addition of specific reagents.Diabetes was induced in rats by intraperitoneal(i.p.) injection of streptozotocin(STZ) at a dose of 65 mg/kg bw.The fasting blood glucose levels were estimated by glucose oxidase-peroxidase reactive strips(Dextrostix,Bayer Diagnostics).Blood samples were taken by cutting the tip of the tail.Serum cholesterol and serum triglycerides were estimated by enzymatic DHBS colorimetric method.Results:Administration of 62.5 and 2S.0 mg/kg of saponins and aqueous extract respectively in normal rats group shows a significant hypoglycemic activity(32.33%and 40.17%respectively) during the first week.However,diabetic group treated with saponin extract produced a maximum fall of 73.1%and 76.03%at day 1 and day 21 compared to the diabetics control.Also,blood glucose levels of the diabetic rats treated with aqueous extract showed decrease of 78.79%on the first day and the effect remains roughly constant during 3 week. Both extracts also declined significantly biochemical parameters(20.77%-49.00%).The control in the loss of body weight was observed in treated diabetic rats as compared to diabetic controls. Conclusions:These results demonstrated significant antidiabetic effects and showed that serum cholesterol and serum triglycerides levels were decreased,significantly,consequently this plant might be of value in diabetes treatment.

Study of total phenol,flavonoids contents and phytochemical screening of various leaves crude extracts of locally grown Thymus vulgaris

Objective:To prepare various crude extracts using different polarities of solvent and to quantitatively evaluate their total phenol,flavonoids contents and phytochemical screening of Thymus vulgaris collected from Al Jabal Al Akhdar,Nizwa.Sultanate of Oman.Methods:The leave sample was extracted with methanol and evaporated.Then it was defatted with water and extracted with different polarities organic solvents with increasing polarities.The prepare hexane,chloroform,ethyl acetate,hutanol and methanol crude extracts were used for their evaluation of total phenol,flavonoids contents and phytochemical screening study.The established conventional metliods were used for quantitative determination of total phenol,flavonoids contents and phytochemical screening.Results:Phytochemical screening for various crude extracts were tested and shown positive result for flavonoids,saponins and steroids compounds.The result for total phenol content was the highest in hutanol and the lowest in methanol crude extract whereas the total flavonoids contents was the highest in methanol and the lowest hexane crude extract.Conclusions:The crude extracts from locally grown Thymus vulgaris showed high concentration of flavonoids and it could be used as antibiotics for different curable and uncurable diseases.

Leaf area development, dry weight accumulation and solar energy conversion efficiencies of Phaseolus vulgaris L. under different soil moisture levels near Nairobi, Kenya

Leaf area development, dry weight accumulation and solar energy conversion efficiencies of Phaseolus vulgaris L. cv GLP\|2 under two soil moisture levels in two contrasting seasons near Nairobi, Kenya were investigated. The experiment confirms that dry weights and yields of Phaseolus vulgaris are limited by a drought induced decrease in leaf area, leading to less radiation interception as a source for assimilation. However, photosynthetic efficiency in Phaseolus vulgaris also appears to decrease and to contribute to these effects. Finally, an even larger decreases of economic efficiency as obtained in the second season, where stress lasted much later into the season, reveals that such a drought also limits considerably the partitioning and translocation of assimilates to the seeds of Phaseolus vulgaris. The efficiencies obtained are in line with the better literature data for other crops.

Essential oil of <i>Thymus vulgaris</i>L. and <i>Rosmarinus officinalis</i>L.: Gas chromatography-mass spectrometry analysis, cytotoxicity and antioxidant properties and antibacterial activities against foodborne pathogens

The essential oil composition of Thymus vulgaris L. and Rosmarinus officinalis L. endemic to France were determined by GC and GC-MS. Oils were assessed for their cytotoxic, antioxidant and antimicrobial activity. 31 and 37 different compounds were identified representing 99.64% and 99.38% of the thyme and rosemary oils respectively, where oxygenated monoterpenes constituted the main chemical class. Thymol (41.33%) and 1.8-cineole (24.10%) were identified as the main constituents of T. vulgaris L. and R. officinalis L., respectively. Essential oils (EOs) of selected plant species were evaluated for their in vitro cytotoxicity against the human lung adenocarcinoma epithelial cell line (A549). Cytotoxicity was measured using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphynyltetra-zolium bromide] colorimetric assay. Dose-dependent studies revealed IC50 of 8.50 ± 0.01 μg/mL and 10.50 ± 0.01 μg/mL after 72 h on the A549 cells for R. officinalis L. and T. vulgaris L., respectively. Antioxidant activity was determined using a quantitative DPPH (1,1-diphenyl- 2-picryl hydrazyl) assay. Thymus and rosemary EOs exhibited effective radical scavenging capacity with 50% inhibitory concentration (IC50) of 437 ± 5.46 μg/mL and 189 ± 2.38 μg/mL respectively and therefore acts as a natural antioxidant agent. The antimicrobial activity of these species has also been studied against several foodborne pathogens and food isolated Salmonella spp. including S. enteritidis of significant importance. According to the results, T. vulgaris L. showed higher bactericidal effect than those from R. officinalis L. These results suggest that the essential oil from T. vulgaris L. and R. officinalis L. have potential to be used as a natural cytotoxic, antioxidant and antimicrobial agent in food processing.

Prunella vulgaris L. extract improves cellular immunity in MDR-TB challenged rats

Objective： To study the effect of the extract of Prunella vulgaris L. on multiple drugs resistant bacillus tuberculosis （MDR-TB）. Methods： Experimental animal model in rats was induced by MDR-TB. Normal group model group and Prunella vulgaris L. group were set up. The contents of IFN-7, IL-4, IL-10 and IL-12 were examined by ELISA. Their genome mRNAs were extracted, the target genes were amplified by PCR. RT-PCR was used to detect the mRNA levels of them. Results： The content of IFN-q, of the extract of Prunella vulgaris L. group was 1.98±0.67 pg/ml, IL-4 was 6.47±1.46 pg/ml, IL-10 was 12.13±3.43 pg/ml and IL-12 was 3.02±0.86 pg/ml. Compared with the model group, Prunella vulgaris L. group was notable difference in serum IFN-γ, IL-12 and IL-10 （P〈0.05）. The mRNA levels of IFN-γ, IL-12 increased and IL-10 decreased obviously, the differences were quite significant （P〈0.05）, but IL-4 had no obvious change. Conclusion： The extract of Prunella vulgaris L. can enhance the cellar immunological function in rats from up-regulation of the level of genetic transcription, accordingly provide the theory basis of healing of tuberculosis with it.

Effect of Macronutrients, Cytokinins and Auxins, on <i>in Vitro</i>Organogenesis of <i>Thymus vulgaris</i>L.

The present study reports an efficient protocol for in vitro propagation of Thymus vulgaris L., an aromatic and medicinal plant in Morocco. Initially, we performed in vitro multiplication of Thymus vulgaris explants existing in the laboratory and obtained from micropropagation by shoot tip culture. Afterwards, we have evaluated the effect of six macronutrients. After that, seven cytokinins (Kin, BAP, 2iP, DPU, Adenine, Zeatine and TDZ) in three different concentrations (0.46, 0.93, 2.32 μM) have been evaluated to optimize cultures multiplication and elongation. Moreover, the effect of three auxins (IAA, IBA and NAA) at 0.57 μM, combined to 4 cytokinins (Kin, BAP, DPU and Ad.) at 0.46 μM, on shoot rooting has been studied. Thereby, MS medium has been proved the most favorable for plantlets growing. Also, we found that the addition of certain cytokinins, specifically 0.46 Kin, 0.46 and 0.93 BAP, 0.46 2iP, 0.46 DPU, 0.46 Ad. and 0.46 Zeat., ensures better multiplication and growth of vitroplants. In addition, multiplication and rooting of cultures were well optimized after addition 0.46 Kin + 0.57 IAA or NAA, 0.46 DPU + 0.57 IBA and 0.46 Ad. + 0.57 IBA combinations to the culture medium. Lastly, plantlets with roots were successfully acclimatized to ex-vitro conditions and these latter served as a source to establish in vitro culture again.

The effects and mechanism of action of Prunella vulgaris L extract on Jurkat human T lymphoma cell proliferation

Objective:The aim of this study was to observe the effect of the Prunella vulgaris L extract on the Jurkat human T lymphoma cell line.Methods:Jurkat cells were cultivated with different concentrations of the extract from Prunella vulgaris L.The MTT assay and flow cytometry were employed to determine the cells' proliferation inhibition ratio and the apoptosis rates,respectively.Agarose gel electrophoresis was used to observe cellular DNA fragmentation,and western blotting was used to observe changes in Bcl-2 and Bax protein expression.Results:The Prunella vulgaris L extract remarkably inhibited the proliferation of Jurkat cells.This inhibition exhibited dose dependence,with an IC50 of 20.23 ± 0.31 μg/mL.Agarose gel electrophoresis showed that the apoptosis strap became wider and brighter,and flow cytometry showed that the apoptosis rate increased in a concentration-dependent manner.Western blotting showed that Bcl-2 protein was down-regulated and Bax protein was up-regulated during apoptosis.Conclusion:The extract from Prunella vulgaris L induced apoptosis of Jurkat cells by down-regulating Bcl-2 protein and up-regulating Bax protein.These actions inhibited the growth of Jurkat cells.

Effects of Different Ridge Culture Ways on Stem Diameter,Plant Height,the Number of Leaves and Yield of Phaseolus vulgaris L. under Dry Farming Conditions

The variety "Aifeng" was used as experimental material,and the stem diameter,plant height,the number of leaves and yield of P. vulgaris under three ways of ridge culture (bedding,high ridge and M ridge) were observed and detected to study the effects of different ridge cultures on the growth and yield. The results showed that the stem diameter,plant height,the number of leaves and yield under M ridge culture were higher than that of bedding and high ridge.

A Metabolomics Study of the Volatile Oil from Prunella vulgaris L.on Pelvic Inflammatory Disease

Objective Pelvic inflammatory disease(PID)is one of the most common gynaecological diseases.Here,this thesis aims to investigate the therapeutic effects of Prunella vulgaris L.oil on the PID by using metabolomics based on gas chromatographymass spectrometry(GC-MS)to address this challenge.Methods First,measurements of pro-inflammatory cytokines and histological analysis of the uterus were conducted to validate the successful generation of a PID rat model.Furthermore,the volatile oil from Prunella vulgaris L.was administered to treat PID rats.Serum samples were collected before and after treatment and analyzed by GC-MS to generate metabolite profiles for each sample.The information generated from the qualitative and quantitative analysis of these metabolites was applied to distinguish between the PID model and normal control groups.Results Some metabolites,such as acetic acid,succinic acid,glyceric acid,(R*,S*)-3,4-dihydroxybutanoic acid,3-hydroxyphenylacetic acid,D-ribose and myo-inositol showed a higher contribution in the classification model;thus,they can be considered as potential biomarkers.Furthermore,the therapeutic effect of the volatile oil extracted from Prunella vulgaris L.could also be visualized using GC-MS-based metabolomics.Conclusions The results show that metabolomics studies are invaluable for disease diagnosis and therapeutic effect estimation.