Study on the Mechanism of Browning of Pomegranate (Punica granatum L.cv.Ganesh) Peel in Different Storage Conditions

Peel browning of pomegranate fruit is the key problem during storage. The mechanism of it during storage in relation to the technology of the browning control was studied in this paper. The effects of storage temperature, gas component and pH value on peel browning of sweet pomegranate fruit in Lintong, Shaanxi Province, China, were investigated. Change in tannin content was measured and the relationship between the activities of ascorbic acid oxidase （AAO）, polypenoloxidase （PPO）, peroxidase （POD）, catalase （CAT） and peel browning was analysed. The results showed tannin was the basic substance of pomegranate peel browning. The activities of the browning index of peel were correlated positively with AAO, PPO and POD, correlated negatively with CAT activity. Application of intermittent warming for 24 h at （15 ± 0.5）℃ every 5 days for pomegranate fruit storage under the conditions of pH 4.0, （5.0± 0.5）℃ and 5.0% CO2 ＋ 8.0% O2 ＋ 87.0% N2 gas component could delay browning, with the browning index being 0.15 after 120 days of storage. Enzymatic tannin denaturation was the main cause that led to peel browning of pomegranate fruit during storage. The browning control of pomegranate fruit peel was effective using intermittent warming storage with suitable pH, temperature and gas component.

Comprehensive evaluation of 20 pomegranate(Punica granatum L.) cultivars in China

Recent investigations on pomegranate products have significantly increased and successfully drawn consumers’ attention to nutritional and medicinal values, promoting the pomegranate industry’s development worldwide. However, little information on pomegranates grown in China is available. Morphological and chemical characterizations of fruits and arils from 20 pomegranate cultivars in six regions of China were investigated. Combined with overall scores by principal component analysis, ‘Yushiliu No. 1’, ‘Taishanhong No. 2’, ‘Tunisia’ and ‘Mollar’ were promising cultivars, and Chinese researchers bred the first two. It was surprising that ‘Mollar’ had bigger fruit size and more aril moisture grown in China than in Spain. Cultivars with higher anthocyanin content in arils were ‘Turkey’, ‘Moyu’ and ‘Red Angel’, which might be used as the source of natural red food colourants. While red husk ‘Hongruyi’ and ‘Hongshuangxi’ with higher vitamin C, aril moisture and lower titratable acid in arils, might also be promising cultivars for further various utilization. Furthermore, the comparison of ‘Tunisia’ fruits from four regions revealed that cultivation locations had more influence on fruit traits than genotypes. Maturity index classification was established for Chinese pomegranate cultivars. Therefore, the results would provide a valuable guide for agricultural cultivation, industrial utilization, and breeding.

Antioxidant Activity of Pomegranate (<i>Punica granatum</i>L.) Fruit Peels

The antioxidant activity of pomegranate fruit peels was evaluated using in vitro tests. 80% methanolic extracts (ME) of peels had higher yield (45.4%) and total phenolics (27.4%) than water (WE) or ether extracts (EE). The reducing power of ME was more potent (P α-catechin. Pomegranate peels contained phenolics, exhibited DPPH scavenging activity and reducing power.

Antinociceptive Effects of Pomegranate (<i>Punica granatum</i>L.) Juice and Seed Extracts on Acute Corneal Pain in Mice

In the present study, the antinociceptive effects of acute (2, 4 and 6 ml/kg) and chronic (1, 2 and 3 ml/kg for 14 days)oral administration of?pomegranate (Punica granatum?L.) juice and seed?extract with or without morphine and naloxane were investigated?on?hypertonic saline-induced acute corneal pain perception in mice. The number of eye wipes with a forelimb was counted for a period of 30 seconds as the criterion for pain assessment. Acute oral administration of the extract?(at 6 ml/kg dose, once) and chronic oral administration?(at 2 and 3 ml/kg for 14 days each) significantly decreased the number of eye wipes after subcutaneous injection of morphine (2 mg/kg, sc), naloxone (2 mg/kg, sc) and normal saline (2 mg/kg, sc) compared with control (p < 0.05). The morphine-induced antinociception was significantly improved by both acute and chronic oral administrations of pomegranate extract (p < 0.05). Naloxone (2 mg/kg, sc) did not reverse the antinociceptive effects of acute (at 6 ml/kg dose, once, oral) and chronic (at 2 and 3 ml/kg for 14 days each) treatments. These findings demonstrate that acute high-dose and long-term lower-dose of pomegranate juice and seed extract can decrease acute corneal pain and improve morphine-induced antinociception in mice.

Correlation of the Cytotoxic and Antioxidant Activities of Moroccan Pomegranate （Punica Granatum） with Phenolic and Flavonoid Contents

The Pomegranate （Punica Granatum）, which belongs tothe Lythraceae family, has been used for centuries in traditional Greco-Arab and Islamic medicine of its vermifuge properties and also to treat various diseases. The aim of this research was to investigate the cytotoxicity and antioxidant activities of Moroccan Pomegranate （peel, leaves, branches, flowers and corolla）. Further, the biological activities were correlated with phytochemical contents of the plant extracts. Methanolic extract from different parts of Punica Granatum was assessed for its antiproliferative activity in two human cancer （breast and colon） cells lines （MBA-MD 231 and HT-29）, through MTT （3-（4,5-dimethyl-2- thiazolyl）-2,5-diphenyl-2H-tetrazolium bromide） bioassay using cell viability and cytotoxicity indices. DPPH （2,2-diphenyl-l-picrylhydrazyl） assay was conducted to screen the antioxidant property of the extracts together with its phenolic and flavonoids content were evaluated, as well. The methanolic extract ofPunica Granatum （peel, leaves, branches, flowers and corolla） showed the highest antiproliferative activity on MBA-MD 231 （IC50 was 133.53-233.32 μg/mL） and HT-29 （IC50 was 127.58-203.24 μg/mL） cells. Antioxidants contents are distributed as follows： peel 〉 leaf 〉 flower 〉 corolla 〉 branches. The inhibitory activities required for decreasing initial DPPH by 50% are 8.27, 9.9, 10.06, 11.67 and 13.28 μg/mL, respectively. These results are in correlation with polyphenols content from corolla, peel, leaves, flower and branches are 120.7, 115, 96.65, 90.73 and 64.67 mg GAE/g dw （mg gallic acid equivalents per g dry weight） and flavonoids are 188.8, 221.7, 180.2, 193.7 and 158.5 mg QE/g dw （mg quercetin equivalents per g dry weight）. Our results show that the peel, flowers, corolla, leaves and branches of Moroccan Pomegranate may contain a lot of bioactive compounds which are responsible for strong antioxidant and cytotoxicity activities observed here. Our finding indicates the possibility of using the extracts of this plant as source of natural antioxidant and anticancer mainly for its abundant phenolic and flavonoid contents.

Determination and Comparison of Pigment Contents in Leaves of Punica granatum L. Hybrids with Different Leaf Colors

[Objective] This study aimed to investigate the reason for the different leaf colors of pomegranate hybrid progeny. [Method] Spectrophotometry was used to determine the contents of three pigments, including chlorophyll, carotenoid and anthocyanin, in the leaves of eight pomegranate individuals and their parents at growth stage. [Result] The results showed that during the early growth stage, the anthocyanin contents of four pomegranate individuals with red leaves were higher than the control, to be specific, the anthocyanin content of No.5 pomegranate individual reached the highest of 4.12 mg/g; the chlorophyll and carotenoid contents of four pomegranate individuals with yellow-green leaves showed consistent dynamic trends with the control, to be specific, the chlorophyll content and chlorophyll/carotenoid ratio of No.4 pomegranate individual reached the highest, while the chlorophyll a/carotenoid b ratio of No.3 pomegranate individual was the lowest. During vigorous growth stage, the contents of chlorophyll, carotenoid and anthocyanin of eight pomegranate individuals showed consistent dynamic trends, with no significant difference with the control. Analysis of variance indicated that there were significant differences in the contents of chlorophyll and carotenoid among different pomegranate individuals at different growth stages and in the content of anthocyanin among different pomegranate individuals, while no significant difference was observed in the chlorophyll/carotenoid ratio among different pomegranate individuals at different growth stages. [Conclusion] The differences in leaf colors among pomegranate hybrid progeny are caused by different contents and ratios of anthocyanin and chlorophyll at growth stage.

Effect of Punica granatum peel extract on learning and memory in rats

Objective:To evaluate potential memory enhancing effect of Punica granatum peel extract on rats.Methods:Healthy adult male albino rats of Wistar strain were used.Each group of 6 rats were administered either distilled water or 50 mg/kg of extract or 100 mg/kg of extract for 15 days and subjected to passive avoidance test or T-maze test.In the next phase rats were administered distilled water or 100 mg/kg of extract for 15 days and the rats were given injection diazepam before subjecting them to the tests.Results:The overall performance was better in test groups compared to control groups.Among the test groups,100 mg/kg rats performed better than 50 mg/kg.The effect on spatial learning parameters like mean number of alternations and mean percentage bias was more marked compared to retention testing parameters like latency. 100 mg/kg Punica extract treated group also improved performance of diazepam treated rats. Conclusions:There is a definite trend of memory improvement by Punica granatum peel with effects being more marked on spatial learning tendency and long term memory than on retention capacity.

Ameliorative properties of Iranian Trigonella foenum-graecum L. seeds and Punica granatum L. peel extracts in streptozotocin-induced experimental diabetic guinea pigs

Objective: To assess the ameliorative properties of Iranian Trigonella foenum-graecum L.(T. foenum-graecum) seeds and Punica granatum L.(P. granatum) peel extracts against streptozotocin-induced diabetes in male guinea pigs.Methods: Sixty guinea pigs were divided into six groups(10 guinea pigs per group).Group 1 consisted of normal animals. Groups 2 and 3 were treated with Iranian T. foenum-graecum seeds and P. granatum peel extract alone, respectively. Group 4 was treated with streptozotocin only; whereas Groups 5 and 6 receiving streptozotocin were treated with Iranian T. foenum-graecum seeds and P. granatum peel extract, respectively.All animals were treated for 30 days, and the body weight, blood and liver biochemical parameters were measured.Results: Guinea pigs exposed to streptozotocin showed an alteration in body weight gain, fasting glucose level, kidney function parameters(blood urea nitrogen and creatinine) as well as decreased serum and hepatic total protein level. In addition, it increased the cholesterol and triglyceride level, while decreasing the hepatic glucose-6 phosphate dehydrogenase activity, glycogen, glutathione content and hepatic catalase activity. Oral treatment with T. foenum-graecum seeds and P. granatum peel extracts revealed significant protective properties with respect to body weight gain and other biochemical parameters studied.Conclusions: The Iranian T. foenum-graecum seeds and P. granatum peel extracts are significantly potent in ameliorating diabetic condition induced by streptozotocin and improving various biochemical parameters in serum and liver of guinea pigs.

Removal of Fluoride from Groundwater by Carbonised <i>Punica granatum</i>Carbon (“CPGC”) Bio-Adsorbent

This study applies the development and application of low cost, Punica granatum bio-adsorbent for the removal of fluoride in groundwater. The batch adsorption study was carried out to analyze the defluoridation by contact time variation, adsorbent dose, adsorbate concentration, adsorbent particle size and presence of co-anions at neutral pH. The analysis of the isotherm equilibrium data using the Langmuir and Freundlich equations by linear methods showed that the data fitted better with Freundlich model (R2 > 0.980). Prepared adsorbent showed enhanced removal of fluoride concentration by 78.1% at equilibrium contact time of 75 minutes. Carbonised Punica granatum Carbon (CPGC) seeds showed a high affinity for fluoride ions compared with other conventional adsorbents. Therefore, it can be considered as a potentially “good”, low-cost bio-adsorbent for de-fluoridation of water compared to other bio-adsorbent.

Phytochemical Determination and Antibacterial Activity of Punica granatum Peel Extracts against Plant Pathogenic Bacteria

Plant pathogenic bacteria are recognized to be harmful microbes able to decrease the quantity and quality of crop production in the world. Punica granatum peel was screened for its potential use as biological control agent for plant pathogenic bacteria. P. granatum peel was successfully extract using n-hexane, methanol and ethyl acetate by maceration. The highest yield obtained by ethyl acetate showed that ethyl acetate extracted more compounds that readily soluble to methanol and n-hexane. For in-vitro antibacterial activity, three different species of plant pathogenic bacteria were used namely Erwinia carotovorum subsp. Carotovorum, Ralstonia solanacearum, and Xanthomonas gardneri. For all crude extracts, four different concentrations 25, 50, 100 and 200 mg/ml were used in cup-plate agar diffusion method. Streptomycin sulfate at concentration 30 μg/ml was used as positive control while each respective solvent used for peel extraction was used as negative control. The results obtained from in vitro studies showed only ethyl acetate extract possessed antibacterial activity tested on the plant pathogenic bacteria. Methanol and n-hexane did not show any antibacterial activity against plant pathogenic bacteria selected where no inhibition zones were recorded. R. solanacearum recorded the highest diameter of inhibition zones for all range of concentrations introduced followed by E. carotovorum subsp. Carotovorum and X. gardneri. For the minimum inhbitory concentration (MIC) and minimum bactericidal concentration (MBC), only the ethyl acetate extract was subjected to the assay as only ethyl acetate extract exhibited antibacterial activity. The minimum concentration of ethyl acetate extract that was able to inhibit plant pathogenic bacteria was recorded at a concentration of 3.12 mg/ml which inhibited R. solancearum and E. carotovorum subsp. Carotovorum, followed by X. gardneri at concentration 6.25 mg/ml. For the minimum bactericidal concentration (MBC), the results showed that at the concentration of 12.5 mg/ml, the extract was still capable of killing the pathogenic bacteria, R. solanacearum, and P. caratovora sub.sp. caratovora while for the bacteria X. gardneri, the concentration that was able to kill the bacteria was 25 mg/ml. The qualitative estimation of phytochemical constituents within P. granatum L. ethyl acetate peel extracts had revealed the presence of tannins, flavonoids, phenols alkaloid, Saponins, and terpenoids. This study has demonstrated that Ethyl Acetate peel extracts of P. granatum has significant antibacterial activity against pathogenic plant bacterial, and it could be of high agricultural value.

<i>In-Vitro</i>Comparison of Antifungal Activity of Herbs (Darehald and Pomegranate) with Azoles

Aim: Candida, an opportunistic organism is one of the commonest causes of hospital acquired infections among fungi. Currently available antifungal drugs have numerous adverse effects and drug-drug interactions (DDIs) along with increase in resistance over the time. Therefore, it is highly emergent to consider alternative treatments for candidal infections, having fewer adverse effects and is cost-effective. The current in-vitro study is undertaken to assess and compare the antifungal effects of the herbs, Berberis aristata (B. aristata, Darehald/Darhald) and Punica granatum (P. granatum, Pomegranate) with fluconazole and voriconazole, based on culture and sensitivity of candidal isolates. Materials and Methods: Ethanolic extracts of herbs (Berberis aristata and Punica granatum) and concentrations were formulated as per standard procedure. 130 samples were obtained for the study from in and out patients reported in clinical subsets of Ziauddin Hospital, Karachi from March to May, 2018. Samples were collected and grown according to the standard procedures like, wet mount test and gram’s staining. Species were identified by CHROM agar candida and API 20 C AUX methods. Sensitivity tests were performed by Kirby Bauer’s disc diffusion method according to CLSI guide lines M-44 A2, 2009. Data analysis was done by one-way ANOVA to compare the antifungal activities of drugs and herbs. Results: Mean inhibitory zones of herbs, B. aristata and P. granatum were highly significant against clinical candidal isolates with respective p-values of 0.00 and 0.02. Both of the herbs, B. aristata and P. granatum were found to be more sensitive, 98.5% and 97.7% respectively in comparison to fluconazole showing 42.3% and voriconazole showing 29.2% sensitivity against candidal isolates. The most resistant candidal specie was C. tropicalis that showed resistance against both fluconazole and voriconazole, contrary to that, this specie was highly sensitive to both of the herbs, showing sensitivity of 100% respective for Darehald and Pomegranate. Conclusion: In comparison to azoles: Culture sensitivity of both herbs (B. aristata and P. granatum) displayed more sensitivity against candidal isolates of patients having non-invasive and invasive candidiasis. These herbs can be considered as substitute or alternative antifungal agents to the conventional antifungal therapy, particularly in cases of treating candidemia patients, which is a life threating condition.

石榴蔗糖代谢相关酶SPS和INV基因家族鉴定与表达分析

蔗糖磷酸合成酶(SPS)和蔗糖转化酶(INV)是蔗糖代谢的关键调控酶,在植物生长发育过程中起重要作用。本研究利用生物信息学和荧光定量PCR等分子手段,鉴定石榴SPS和INV基因家族成员,分析其理化性质、保守结构域、保守基序、二级结构、亚细胞定位、系统进化关系和表达模式。结果表明,从石榴基因组中鉴定出4个SPS基因和11个INV基因,其编码蛋白均为不稳定蛋白,具有典型的保守结构域,家族成员间特征motif数量和种类大致相同,蛋白结构高度保守;这些蛋白不均匀地分布在染色体上,均定位于叶绿体中,二级结构主要由α-螺旋和无规则卷曲组成。石榴SPS和INV基因家族成员间存在不同程度的亲缘关系,与巨桉同源性较高;不同SPS和INV基因在石榴果实不同发育时期的表达模式存在差异,PgINV3在9月15日(果实增大期)表达水平最高,显著高于其他时期。本研究结果对解析石榴果实中蔗糖代谢的分子机理具有重要意义。

纤维素酶辅助提取石榴籽饼粕多糖工艺优化及其动力学、热力学研究

目的优化纤维素酶辅助提取石榴籽饼粕多糖工艺,并考察其动力学、热力学。方法在单因素试验基础上,以加酶量、介质pH、提取温度、提取时间为影响因素,多糖得率为评价指标,Box-Behnken响应面法优化提取工艺,计算动力学、热力学参数。结果最佳条件为加酶量0.9%,介质pH 4,提取温度49℃(322 K),提取时间2.6 h,多糖得率为5.89%。温度为322 K时,实验组k为0.0191 min^(-1),D s为4.843×10^(-6)cm^(2)/min,高于对照组。在307~317 K的提取过程中,焓ΔH为156.378 kJ/mol,熵ΔS为520.1904 J/(mol·K),为吸热熵增加;在327~337 K的提取过程中,焓ΔH为-147.931 kJ/mol,熵ΔS为-428.5784 J/(mol·K),为放热熵减小;自由能ΔG值均小于0,为自发过程。结论该方法稳定可靠,可用于纤维素酶辅助提取石榴籽饼粕多糖的工程放大和优化控制。

新疆石榴表型性状分析及分子身份证构建

为快速鉴别石榴种质资源,解决石榴“同名异物”、“同物异名”现象,促进新疆石榴产业的发展,本研究在采集新疆石榴主产区24份种质的基础上,分析新疆石榴种质叶片及果实表型性状的变异特征,选择多态性较好的18条引物对石榴种质进行遗传多样性分析及亲缘关系鉴定,并从中选出可将24份石榴种质鉴别出来的核心引物构建新疆石榴的分子身份证。结果表明:新疆石榴的果实、叶片形状变异较大,遗传多样性较为丰富,但仅从表型性状很难将石榴种质进行精准鉴别。18条引物共扩增出240条条带,其中多态性条带199条,占比为82.92%。24份种质的平均等位基因数为1.812 5,有效等位基因数为1.287 0,Nei’s基因多样性指数为0.177 0,Shannon’s多样性指数为0.282 0;和田地区石榴种质的Nei’s基因多样性指数及Shannon’s多样性指数高于喀什地区石榴种质。在遗传相似系数为0.85时,24份石榴种质被分为6大类群,且石榴的分类结果与果实风味和地理位置的关联性不大。5条核心引物UBC73、UBC880、UBC810、UBC841、UBC35可实现24份石榴种质的鉴别,以其扩增的电泳图谱所构建的新疆石榴的分子身份证也互不相同,表明24份种质不存在“同名异物”、“同物异名”现象。本研究结果为石榴种质鉴定提供了一种新方法。

响应面法优化甜石榴皮生物碱提取工艺及其抗氧化活性评价

以甜石榴皮为原料,采用超声波辅助法提取生物碱。以生物碱得率为指标,在单因素试验的基础上,通过响应面法优化甜石榴皮生物碱的提取工艺,并评价其抗氧化活性。结果表明:最佳提取工艺为乙醇体积分数32%、超声时间51 min、超声温度39℃、料液比1∶17(g/mL),在此条件下生物碱得率为2.35%。抗氧化试验表明,在一定质量浓度范围内,甜石榴皮生物碱对DPPH自由基和羟基自由基的清除率最高分别为82.32%、64.36%,IC50分别为0.368、0.590 mg/mL,具有较好的抗氧化活性。

基于网络药理学预测石榴皮抗甲型流感机制及初步药效学验证

目的:基于网络药理学预测石榴皮抗甲型流感的潜在有效靶点及作用机制,并通过动物实验进行验证。方法:通过TCMSP技术平台检索石榴皮的有效成分,筛选其作用的靶标。甲型流感的靶标通过TTD、DRUGBANK等数据库筛选。利用Cytoscape绘制石榴皮抗甲型流感网络图,通过STRING数据库构建蛋白互作网络,并对关键靶标的生物功能进行注释与通路分析。甲型流感病毒感染小鼠,使用酶联免疫吸附试验(ELISA)、蛋白质印迹法(Western blot)检测白细胞介素17(interleukin 17,IL-17)通路中关键蛋白、炎症因子的表达,观测小鼠体质量、肺指数变化。结果:石榴皮中有7个潜在有效成分抗流感病毒、96个有效靶点,其中RELA、MAPK1、JUN、HIF1A、ESR1、NFKBIA、TP53可能为核心靶点,其作用机制可能与IL-17信号通路、化学致癌-受体激活、磷脂酰肌醇3-激酶等通路有关。初步的动物实验显示,石榴皮提取物参与调控IL-17通路,抑制炎症因子白细胞介素6(interleukin 6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素1β(interleukin 1β,IL-1β)、TRAF6(TNF receptor associated factor 6)及核因子κB(nuclear factor kappa-B,NF-κB)p65蛋白表达(P<0.05)。结论:通过预测得出石榴皮中所含的6个成分与不同靶点、通路交互作用,抑制多个炎症因子相关蛋白的表达,进而防治甲型流感。

山东石榴品种遗传多样性与亲缘关系的荧光AFLP分析

利用荧光AFLP标记技术,采用8对EcoR I/Mse I引物对山东省25个石榴品种进行AFLP-PCR分析,结果表明:25个石榴品种扩增的多态性位点数从70条到126条不等,平均90.25条,平均多态性位点41.78%;所有检出位点的平均观测等位基因数、平均有效等位基因数、平均Nei's基因多样度和平均香农信息指数分别为1.4178、1.1874、0.1133和0.1752;方差分析显示各位点的Nei's基因多样度和香农信息指数具有不同程度的显著性差异;基于Lei&Li相似系数利用UPGMA法进行聚类分析,在相似系数0.786处将25个品种划分为4大类。根据AFLP结果探讨了山东石榴品种的亲缘关系与新品种选育的前景。

云南25份石榴资源的RAPD分析

用RAPD标记技术分析了25份云南地方石榴材料(品种或类型)的亲缘关系,从128个随机引物中筛选出12个重复性好、条带清晰的多态性引物,并用这些引物对25份云南石榴材料进行RAPD扩增,共扩增出110条谱带,平均每个引物9.17条。多态性程度达到71.8%。用Statistic分析软件计算出的遗传距离为0.027～0.342,对25份材料进行了UPGA法聚类,亲缘关系树状图显示,25份云南石榴材料的遗传背景较复杂,难以划分类群。研究结果与生产上根据风味、花色、皮色、子粒颜色、核软硬程度等某一性状及栽培目的的分类方法不一致。

石榴离体培养再生体系的研究

对石榴休眠枝段、成熟叶片及当年生新梢离体培养 ,建立其再生体系。结果表明 ,以休眠枝段、成熟叶片为外植体均能形成愈伤组织并分化出不定芽 ;当年生新梢茎段培养 ,以MS +BA 2 0mg·L-1+NAA0 3mg·L-1对茎段腋芽的增殖最适宜。诱导生根 ,用 1/2MS为基本培养基附加NAA 0 5mg·L-1+活性炭0 1mg·L-1+蔗糖 2 0g·L-1,生根率达 95 8%。培养基中附加 0 1%活性炭 ,对促进生根均有显著效果。

中药石榴皮鞣质成分抗生殖器疱疹病毒作用

采用细胞培养技术，证实鞣质是石榴皮抗生殖器疱疹病毒的有效成分。其特点不仅能够抑制病毒在细胞内的增殖，更为重要的是显示较强的直接杀灭病毒和阻止其吸附细胞的作用。