Free radical scavenging potential of Lagenaria siceraria(Molina) Standl fruits extract

Objective:To elucidate free radical scavenging activity of ethanolic extract Lagenaria siceraria （L.siceraria）（Molina） fruit.Methods:The free radical scavenging activity of the L siceraria （Molina） fruit extract was assayed by usingα,α-diphenyl-β-picrylhydrazyl（DPPH）. 2,20-azinobis 3-ethyl henzothiazoline-6-sulfonate（ABTS）,FRAP,reducing power,chelating ability andβ-carotene bleaching assay.Results:The IC50values of DPPH and ABTS radicalscavenging activity was found to be 1.95 mg/mL and 19 mg/mL,respectively.In ferrous chelation assay,the percentage of inhibition was found to be 89.21%.The reducing power of ethanolic extract of L siceraria（Molina） fruit was 0.068 at 1 mg/mL and increased to 0.192 at 5 mg/mL Theβ-carotene linoleate bleaching assay was 46.7%at 5 mg/mL and antioxidant activity using FRAP at 0.305 for 1 mg/mL to 0.969 for 5 mg/mL Conclusions:The results indicate that L siceraria （Molina） fruit could be an important sources of natural radical scavengers.

Lipase inhibitory activity of Lagenaria siceraria fruit as a strategy to treat obesity

Objective:To explore pancreatic lipase inhibitory activity under different extraction conditions in order to track the most potent extract.Methods:The methanolic extract and its fractions in solvents of increasing polarity,ether,chloroform,ethyl acetate,n-butanol and water,were made through cold maceration.Extracts in ethanol,ethyl acetate,acetone and chloroform were similarly prepared.Aqueous extract was prepared through hot decoction method.A reported method was used to determine lipase inhibitory activity of extracts and fractions over wide ranges of concentrations.Results:The extracts and fractions exhibited concentration dependent activity.The IC50(μg/mL)values of methanolic,ethanolic,chloroform,ethyl acetate,acetone,ethyl acetate(after washing with water)and aqueous decoction were 293.40,266.47,157.59,182.12,352.34,257.00,and 190.00,respectively.The activity of chloroform,ethyl acetate and aqueous extracts were close to that of the drug orlistat(IC50 146μg/mL).Out of the fractions of the methanolic extract,the chloroform fraction was most active(IC50 189.6μg/mL).The order of inhibitory activity of the fractions was as follows:chloroform>ether>n-butanolic>aqueous>ethyl acetate.The GC/MS analysis of the most active chloroform faction showed the presence of hexadecanoic acid,methyl hexadecanoate,isopropyl palmitate,methyl 9,12-octadecadienate,and methyl 9,12,15-octadecatrienoate.Conclusions:The study suggests that Lagenaria siceraria has potential to inhibit pancreatic lipase activity,suppressing lipid digestion and thereby diminishing entry of lipids into the body.Regular intake of aqueous decoction of the fruit may therefore be recommended for control of obesity.Fatty acids and their esters may play role as inhibitors of lipase.

Phytogeographical Distribution and Fruit Diversity of <i>Lagenaria siceraria</i>Species in Nigeria

Phytogeographical distribution and fruit diversity of Lagenaria siceraria (Mol.) Standl species in Nigeria were accessed. Information on the distribution and fruit diversity was sourced from three herbaria, Forestry Research Institute of Nigeria Herberia, University of Port Harcourt Herberia and the University of Ibadan Herberia. A field survey across fifteen states in Nigeria namely Kaduna, Kano, Plateau, Oyo, Akwa Ibom, Cross River, Taraba, Bayelsa and Enugu, Ebonyi, Ondo, Ogun, Kwara Anambra and Rivers State. A total of twenty-four fruit shapes were accessed. The diversity of fruit shapes across the geographical regions of Nigeria shows that the Guinea savannah has all the twenty-four different landraces represented, while Tropical rain forest has nineteen, Sudan savannah has seventeen and Freshwater swamp and the Saltwater swamp has eleven different fruit shapes respectively. The fruits of Lagenaria siceraria (Mol) Standl is considered a topocline since it is not affected only by environmental factors but possess a wide distribution pattern across the geographical regions of Nigeria.

Transcriptome based high-throughput SSRs and SNPs discovery in the medicinal plant Lagenaria siceraria

Lagenaria siceraria(Molina)Standley has unique biological characteristics with high nutritional and medicinal values.It is an important pharmaceutical plant with various biologically active ingredients.Genetic improvement and deeper genomic studies require a rich resource of molecular markers.The application of next-generation sequencing technology,especially for transcriptome profiling,has greatly facilitated high throughput single nucleotide polymorphism(SNP)and simple sequence repeat(SSR)discovery.In this study,we sequenced the transcriptome of three major cultivars of L.siceraria and obtained 64.88 GB of clean data.The assembled high-quality reads were clustered into 89,347 unigenes,which were annotated by non-redundant protein database,Swiss-Port,Eukaryotic Ortholog Groups,Kyoto Encyclopedia of Genes and Genomes,and gene ontology databases.A total of 8,891 SSR and 35,873 SNP markers were predicted from unigenes by MISA and SAM tools,respectively.Characterization of the predicted markers in L.siceraria showed that the SSR and SNP densities were 60 and 243 markers per Mb of genome,respectively,and the estimated ratio of transition to transversion of SNP was 2.016.These markers will be very useful for genetic studies in L.siceraria,especially for the high-density linkage map construction and genome-wide association studies.Further genomic studies based on these results will facilitate the identification of novel genes or alleles of pharmaceutical importance.

Purification, characterization, and bioinformatics studies of phosphatidic acid phosphohydrolase from <i>Lagenaria siceraria</i>

Phosphatidic acid phosphohydrolase (PAP), EC 3.1.3.4, is the penultimate step in the Kennedy pathway of triacyl glycerol (TAG) synthesis leading to the formation of diacylglycerol (DAG), which is a key intermediate in TAG synthesis. We partially purified a soluble PAP from mid maturing seeds of bottle gourd, Lagenaria siceraria. The steps include both anionic and cationic ion exchanger columns. Catalytic characterization of the partially purified PAP revealed that the optimum pH and temperature for activity were at 5.5?C and 45?C. Under optimum assay condition using dioleoyl phosphatidic acid (DPA) as the substrate, the Vmax and Km were 0.36 ηkat/mg of protein and 200 μM, respectively. For the synthetic substrate, ρ-nitrophenylphosphate, ρ-NPP, the Vmax and Km were 33.0 nkat/mg of protein and 140 μM, respectively. The activity was neither inhibited nor enhanced by the presence of Mg2+ at a concentration range of 0 to 10 mM. Two major protein bands at 42-kDa and 27-kDa were visible in SDS-PAGE after partial purification. Bioinformatics analysis of tryp-sinized protein fractions containing PAP activity showed peptide sequences with sequence homology to various phosphate metabolizing enzymes including cucumber and castor bean purple acid phosphatase, polyphosphate kinase, fructose biphosphate aldolase, and enolase from various dicotyledonous plants including rice, corn, grape, and Arabidopsis lyrata.

Determination of the Genetic Structure of the Oleaginous <i>Lagenaria siceraria</i>of the Nangui Abrogoua University Germplasm Collection

Thirty accessions of Lagenaria siceraria from the Nangui Abrogoua University germplasm collection were analyzed using three microsatellite markers. The average Polymorphism Information Content (PIC) value was 0.61. The average observed heterozygosity value (H0 = 0.631) compared to the average expected heterozygosity value (He = 0.645) did not show significant differences in the selected accessions, which suggested random mating in the set of accessions. Within accession inbreeding estimate (FIS), was 40% and was not significantly different from zero. The reduction of heterozygotes was likely the result of presence of null alleles. Analysis of Molecular Variance (AMOVA) within and among 30 accessions of L. siceraria revealed that 39% of the total variation resides among accessions and 61% within accessions. Accession structuring pattern derived from Bayesian clustering analysis revealed two clusters. Based on the genetic structure of the accessions analyzed, a sampling strategy to collect and conserve genetic resources of L. siceraria was suggested.

Bioinformatics a Tool for Biosystematics Studies of <i>Lagenaria siceraria</i>(Mol.) Standl. Complex

Bioinformatics has been a major tool in the revolution of plant systematics in recent times. The diversity of fruit shapes in Lagenaria siceraria (Mol.) standl. species has been of great concern because of its fruit complexity. This study is based on the application of rubisco enzyme using rbcL marker because of its conservativeness and its ability to discriminate below the specific level hence its usage to sequence the chloroplast genome with ABI, PRISM 377 DNA sequencer. The sequences obtained were viewed using MEGA X software and subsequently subjected to validation through National Center for Biotechnology Information (NCBI) using Nucleotide Basic Local Alignment Search Tool (BLAST N). The result obtained showed that all the sequences belong to Lagenaria siceraria (Mol.) standl. with percentages ranging from 95% to 100% for query cover sequences and 98% to 100% for identity sequences. From the taxonomic report obtained sequence A, C, G, H, J, Q has the highest hits of 44 on Lagenaria siceraria out of 109 total value, sequence O and R has the highest hits of 44 on Lagenaria siceraria out of the total value of 111, sequence V has the highest hits of 44 on Lagenaria siceraria out of 119 total value and sequence X finally has the highest value hits of 44 on Lagenaria sceriania out of 105 total value, based on this report, phylogenetic tree was constructed to show the level of relatedness of the different fruit diversity of L. siceraria complex. This work therefore has aided in the molecular characterization of Lagenaria siceraria (Mol.) standl. landraces found in Nigeria.

Review of Research Situation of Lagenaria siceraria(Molina) Standl.

Lagenaria siceraria( Molina) Standl. Var. hispisa( Thund.) Hara is an annual creeping herb of the family Cucurbitaceae. It has high nutritional value and medicinal value,and it is a high-quality vegetable with low sugar and many kinds of vitamins. This paper introduced the current research situation of L. siceraria( Molina) Standl. from medicinal value,cultivation management technology and processing technology. Besides,it discussed main points of future researches on the basis of existing research,to promote the development of L. siceraria( Molina) Standl.

Genetic Architecture of Yield in Bottle Gourd (<i>Lagenaria siceraria</i>(Mol.) Standl.)

The genetic architecture of yield was studied in a five parent half diallel cross of bottle gourd at the experimental field of Olericulture Division, Horticulture Research Centre (HRC), Bangladesh Agricultural Research Institute (BARI), Gazipur during the winter season of 2016-17. The values of mean square for GCA (general combining ability) and SCA (specific combining ability) were highly significant which suggested the presence of both additive and non-additive genetic variance in the population. But the higher magnitude of GCA compared to SCA indicated predominance of additive genetic variance. In most of the cases, the cross between poor and poor parents showed positive SCA effect for fruit yield, which indicated the higher yield. The estimates of mid parent heterosis ranged from &#8722;19.0 to 31.8 percent and the better parent heterosis ranged from &#8722;28.4 to 20.5 percent. Analysis for genetic components of variation suggested that additive components were more important in the inheritance of fruit yield. This character was observed being controlled by two to three pairs of genes or groups of genes. Narrow sense heritability was 55 percent indicating probability of selection in generations. The graphical analysis also indicated wide genetic diversity among the parents.

Antioxidant Activity and Anti-Hyperglycemic Effect of Lagenaria Siceraria Fruit Extract

Lagenaria siceraria （Molina） Standl., known as bottle gourd is widely available throughout Vietnam as an edible vegetable. Bottle gourd is a commonly fruit but there are little reports about its biological activities. The aim of the present study was to evaluate antioxidant activity and anti-hyperglycemic potential of ethanolic extract of bottle gourd. The ethanolic extract was tested for its antioxidant activity on chelating power, the ferric reducing/antioxidant power assay （FRAP） and the 1, 1-diphenyl-2-picryl-hydrazyl （DPPH） free radical scavenging assay. The results showed that chelating activity was 80.02%, FRAP value was 92.84 μmol Fe^2＋/g and active radical scavenging which showed an IC50 was 49.50 μg/ml. The five different doses of ethanolic extract were treated by oral tolerance to mice body before oral glucose tolerance and blood glucose level was measured using glucose oxidase method. The result indicated that inhibition percentage of ethanolic extract of bottle gourd at the dose of 400 mg/kg in glucose-induced hyperglycemic mice was effectively similar to standard drug glibenclamide. The results obtained indicate that ethanolic extract from bottle gourd is the potential antioxidant and anti-hyperglycemic agent.

瓠瓜(Lagenaria siceraria)白粉病抗性的全基因组关联分析

白粉病是瓜类作物重要病害之一。本研究采用苗期人工接种方法,研究了117份瓠瓜微核心种质对白粉病的抗性,并基于不同材料的病情指数和基因型数据,进行全基因组关联分析(genome-wide association analysis,GWAS)。利用2014年和2018年的抗病表型数据,分别鉴定到22个和13个与白粉病抗性相关的SNPs,其中2个SNPs位点在两年试验中同时被检测到。进一步研究发现不同瓠瓜材料的白粉病抗性与其携带有利等位变异数目成正相关。本研究为克隆瓠瓜抗白粉病相关基因、研究其抗病机制提供科学依据,促进了瓠瓜抗白粉病分子育种。

瓠瓜CAT家族成员鉴定及响应白粉病菌的表达分析

为探究瓠瓜(Lagenaria siceraria)CAT基因家族的特性及在白粉病菌侵染后的作用,基于瓠瓜基因组数据和转录组数据,对LsCATs家族成员进行了鉴定与分析,研究其表达模式。结果表明:在瓠瓜基因组中共鉴定到2个CAT家族同源基因,其编码蛋白氨基酸数分别为489、1019个,且均为亲水性蛋白;进化分析表明,瓠瓜CAT基因家族被分成2个亚组,推测在相同亚组的瓠瓜与黄瓜CAT转录因子可能具有相似的功能;利用MEME软件预测得到15个motif元件,不同CAT基因家族的motif数量及分布存在一定差异;顺式作用元件分析表明,LsCATs的启动子包含ABA、乙烯、MeJA、干旱、低温等多种与逆境胁迫有关的响应元件;瓠瓜接种白粉病菌后,LsCAT1表达量呈现出先升高后降低的趋势,LsCAT2则表现为先下降后升高的趋势,表明LsCAT1、LsCAT2可能与瓠瓜的白粉病抗性相关。本研究可为CAT基因在瓠瓜抗病机制的研究提供理论依据。

Inhibition of ADP-induced platelet aggregation and involvement of non-cellular blood chemical mediators are responsible for the antithrombotic potential of the fruits of Lagenaria siceraria

AIM: The fruits of Lagenaria siceraria(Molina) Standl.(Cucurbitaceae), a commonly used vegetable, are reported to possess various medicinal properties. In previous studies, the fibrinolytic potential of an ethanolic extract of fruits of Lagenaria siceraria was investigated in comparison with kaempferol isolated from it. The aim of the present study was to explore its mechanistic antithrombotic potential and antiplatelet activity using a wide dose range in different in vitro and in vivo models, and to quantify the total phenolic, flavonoid, and kaempferol contents using a colorimetric method. METHOD: The antithrombotic potential was investigated using tail bleeding time in mice, a plasma recalcification assay, and pulmonary thromboembolism in mice. The antiplatelet activity was studied using an in vitro model to investigate IC50 value. RESULTS: A significant amount of total phenols, flavonoids, and kaempferol was quantified in L. siceraria ethanolic extract. An ethanolic extract of the fruits of L. siceraria showed a significant increase in tail bleeding time and plasma recalcification time, significant protection against ADP induced pulmonary thromboembolism in mice, and also inhibited the platelet aggregation induced by ADP in vitro. The study suggested that the fruits of L. siceraria exhibit significant antithrombotic potential due to inhibition of ADP-mediated platelet aggregation and the involvement of various non-cellular chemical mediators of blood. CONCLUSION: This finding may be helpful in treating the serious consequences of the thrombus formed in blood vessels which include atherothrombotic diseases, such as myocardial or cerebral infarction. So, further investigation should be done for revealing exact mechanism of action behind these types of activities.

STUDY ON MECHANISM OF ETHYLENE CONTROL OF SEX DIFFERENTIATION IN Lagenaria siceraria

The sex expression of Lagenaria siceraria can be modified by phytohormones. This modification is related to "a certain bisexual stage" during floral bud differentiation. ACC (ethylene precursor) and STS (ethylene antagonist) can change the direction of sex differentiation of the floral buds in vitro. STS induces maleness by means of inhibiting ethylene action, directly promoting the development of the primordia of stamens, and then suppressing that of pistils. Hence, it can be deduced that ethylene induces femaleness by directly suppressing the development of the primordia of stamens and then promoting that of pistils. Such a regulation of the development of sexual organs is due to a change in the direction of assimilates to the pistils by ACC and to the stamens by STS. Therefore, ethylene induces femaleness by first limiting the sink strength of the stamens and then increasing that of the pistils.

Moisture dependent physical properties of Lagenaria siceraria seed

The investigation was done to evaluate the effect of moisture content on some physical properties of Lagenaria siceraria seed.The study was conducted at five moisture levels including 8.84%,10.10%,11.89%,12.60%and 15.10%(w.b.).Results showed that the mean value of principal dimensions,average diameters,surface area and 1000 seed mass increased linearly but aspect ratio decreased with increase in moisture content.The sphericity increased in the moisture range of 8.84%to 11.89%but decreased with further raise in the moisture up to 15.10%.Gravimetric properties like bulk density increased with increase in moisture content however true density and porosity decreased linearly with increase in moisture content.Angle of repose and terminal velocity increased linearly with moisture content of the seed.The coefficient of friction increased linearly with seed moisture content on five experimental surfaces(plywood,galvanized iron,glass and plastic).The information pertaining to moisture dependant physical properties of Lagenaria siceraria seeds may become an essential part in design of processing machines and its unit operations,design of dehulling,oil expression and other processing equipments.

不同盐浓度对观赏葫芦幼苗生长及生理特性的影响

以观赏葫芦幼苗为试材,试验研究了不同盐浓度(0、50、100、200、300 mmol/L NaCl)对观赏葫芦幼苗生长及生理特性的影响。结果表明:当盐浓度为50 mmol/L(即S 50处理)时,幼苗受到的危害较小;随着盐浓度的不断提高,当盐浓度为300 mmol/L,盐胁迫会抑制观赏葫芦幼苗的株高(下降17.70%),减少幼苗生物量的积累,叶绿素总含量降低26.54%,降低幼苗抗氧化酶(超氧化物歧化酶、过氧化物酶、过氧化氢酶和抗坏血酸过氧化物酶)活性,增强幼苗体内抗性指标(脯氨酸、丙二醛和可溶性糖)含量的积累以及植物细胞膜选择透过性。当NaCl溶液在200 mmol/L及以下范围,观赏葫芦幼苗会通过调控体内渗透物质等减轻胁迫伤害;当NaCl溶液在200 mmol/L以上范围,观赏葫芦幼苗的叶片细胞结构受损严重进而影响生长。为此,建议在盐溶液为200 mmol/L以下范围的区域试种及推广种植。

38份瓠瓜种质资源遗传多样性的ISSR分析

采用ISSR分子标记技术对源自中国7个省份的38份瓠瓜种质进行遗传多样性分析。12个ISSR引物共扩增出96条多态性带,平均每个引物扩增的多态性带数为8条,多态性比率平均为83.5%。聚类分析将供试的38份种质分为4个类群8组,主坐标分析将其分为4个类群10组。ISSR分子标记的分类结果与瓠瓜的农艺性状分类和地理分布有一定的相关性。

不同产地瓠瓜品种ITS序列的遗传多样性分析

对国产29个瓠瓜也Lagenariasiceraria（Molina）Standl.页品种的ITS序列进行了扩增及测序,并结合引自GenBank的国产9个瓠瓜品种以及国外6个瓠瓜品种和3个同属种类的ITS序列,对它们的ITS序列长度和GC含量以及变异位点进行比较,在此基础上构建系统发育树并对47个样本间的遗传关系进行研究。结果显示：供试47个样本的ITS序列均由ITS1、5.8SrDNA及ITS2组成,各样本间的ITS序列长度、GC含量以及变异位点差异明显。国产38个瓠瓜品种的ITS序列（包括ITS1、5.8SrDNA及ITS2）长度为619-627bp、GC含量为58.00%-63.32%；国外9个样本的ITS序列长度为591-626bp,GC含量为54.17%-63.26%。序列比对结果显示：国产38个瓠瓜品种的ITS序列同源率为84.6%-100.0%,包含221个变异位点；其中,来源于山东的品种‘砧木2’（‘ZhenmuNo.2’）的ITS序列包含的变异位点最多,与其他品种间的同源率也最低。在系统发育树上,国产38个瓠瓜品种可分为3个分支,来源于山东的品种‘砧木2’和来源于河南的品种‘西瓜砧木1’（‘XiguazhenmuNo.1’）各自聚为第1和第2分支；其余36个品种聚为第3分支。而供试的47个样本则可分为2个分支和5个亚组,第1分支可分为2个亚组,包括国产品种‘砧木2’和产自日本的2个品种；第2分支包含的44个样本则进一步分为3个亚组,国产品种‘西瓜砧木1’和产自法国的品种‘白花瓠瓜’（‘White-floweredgourd’）各自聚为第1和第2亚组,其余的42个样本聚为第3亚组。研究结果表明：供试的不同产地瓠瓜品种间存在丰富的遗传变异和地理分化现象,其ITS序列差异与地理分布有一定关系。

基于NBS-LRR类R基因保守结构域克隆瓠瓜抗病基因同源序列

【目的】分离鉴定瓠瓜Lagenaria siceraria抗病种质的抗病基因同源序列（Resistance gene analogs,RGAs）,为瓠瓜功能性抗病基因的克隆及分子标记辅助育种奠定基础.【方法】根据已知核苷酸结合位点和富亮氨酸重复（Nucleotide binding site and leucine rich repeat,NBS-LRR）类抗病基因保守区设计简并引物,从“大籽瓠”抗性材料基因组DNA中分离抗病基因同源序列,并利用生物信息学软件进行长度变异、保守结构域、同源比对与系统进化分析.【结果和结论】基于同源克隆策略,获得23条瓠瓜NBS抗病同源序列,命名为HNB1～HNB23,GenBank登录号为KJ908192～KJ908214.序列分析及同源比对结果表明,这些RGAs长度变异在242～261nt之间,18条序列具有连续开放阅读框（Open reading frame,ORF）,推导氨基酸序列具有P-loop、Kinase-2a典型NBS类R基因保守结构域;核苷酸序列相似性为41.5% ～98.8%,氨基酸序列相似性为21.5% ～100.0%;利用NCBIBlast同源搜索发现,与其他植物尤其是冬瓜、黄瓜、葫芦及丝瓜的已知R基因具有40% ～100%相似性;氨基酸序列聚类分析将其分为5个组;同源进化分析表明,23条瓠瓜RGAs均为non-TIR-NBS-LRR类R基因,与推导氨基酸序列多重比较结果一致.

十种植物生长调节剂处理对苦瓜和葫芦种子萌发的影响

以苦瓜和葫芦种子为试材,设定10种植物生长调节剂的不同浓度(0、1、2、5、10、50mg·L^(-1))梯度,采用纸床法,分别测定10种植物生长调节剂对苦瓜和葫芦种子萌发的影响,以期获得10种植物生长调节剂对苦瓜和葫芦种子萌发的最佳浓度。结果表明:复硝酚钠(CSN)、α-萘乙酸钠(NAA)、吲哚丁酸钾(IBA-K)、胺鲜脂(DA-6)、6-苄氨基嘌呤(6-BA)、赤霉素(GA_3)、6-糠氨基嘌呤(KT)、3-吲哚乙酸(IAA)、3-吲哚丁酸(IBA)处理浓度分别为5、2、10、10、2、5、5、5、5mg·L^(-1)时苦瓜和葫芦种子的发芽率、发芽势和发芽指数均最佳,而氯吡脲(CPPU)不同浓度处理均抑制苦瓜和葫芦的种子萌发。建议使用CSN、NAA、IBA-K、DA-6、6-BA、GA_3、KT、IAA、IBA处理苦瓜和葫芦种子时浓度分别设定为≤5、≤5、≤10、≤10、≤5、≤5、≤10、≤5、≤5mg·L^(-1)。