Xanthomonas axonopodis pv. punicae uses XopL effector to suppress pomegranate immunity

Xanthomonas axonopodis pv. punicae （Xap） causing bacterial blight is an important pathogen that incurs significant losses to the exportability of pomegranate. Xap uses the Xop TTSS-effector, via the type three secretion system, to suppress pomegranate immunity. Here, we investigate the role of XopL during blight pathogenesis. We observed that XopL is essential for its in planta growth and full virulence. Leaves inoculated with Xap AxopL produced restricted water-soaked lesions compared to those inoculated with wild-type Xap. XopL supports Xap for its sustained multiplication in pomegranate by suppress- ing the plant cell death （PCD） event. We further demon- strated that XopL suppresses immune responses, such as callose deposition and production of reactive oxygen species （ROS）. RT-qPCR analysis revealed that immune responsive genes were upregulated when challenged with Xap AxopL, whereas upregulation of such genes was compromised in the complemented strain containing the xopL gene. The transiently expressed XopL：：EYFP fusion protein was local- ized to the plasma membrane, indicating the possible site of its action. Altogether, this study highlights that XopL is an important TTSS-effector of Xap that suppresses plant immune responses, including PCD, presumably to support the multiplication of Xap for a sufficient time-period during blight disease development.

Antiparasitic and apoptotic modulatory activities of curcumin and extracts of Nigella sativa L,Zingiber officinale Rosc.,and Punica granatum L.in combination with spiramycin against chronic cerebral toxoplasmosis in immunocompromised mice

Objective:To examine the effect of coadministration of different medicinal plant extracts(curcumin and extracts of Nigella sativa L.[N.sativa,Jia Hei Zhong Cao],Zingiber officinale Rosc.[Z.officinale,Sheng Jiang],and Punica granatum L.[P.granatum,Shi Liu])with spiramycin(SP)against the cystogenic ME-49 Toxoplasma gondii(T.gondii)strain in immunocompromised mice.Methods:We utilized 68 mice categorized into 8 groups:2 non-infected controls(immunocompetent and immunocompromised),1 infected control,and 5 infected and treated groups.Following the experiment,the cerebral tissues of each mouse underwent parasitological,histopathological,and immunohistochemical evaluations.Results:Compared with the infected non-treated group,all infected treated groups showed significant reductions in brain cyst numbers(all P<0.05),with the highest reduction rate(77.4%)recorded for P.granatum and SP combination(G8).Only G8 showed a significant reduction in mouse deaths compared with the other groups.With regards to restoring histopathological changes and decreasing inflammation,the groups infected and treated with curcumin and P.granatum combined with SP showed the best results(P<0.05).Combinations of curcumin and extracts of Z.officinale and P.granatum with SP signif-icantly restored the cerebral expression of caspase-3 compared with the N.sativa extract combination(P<0.05).G8 showed non-significant expression compared with the local expression in the negative control groups.Conclusion:Our study revealed that the coadministration of P.granatum extract with SP was the most effective combination against chronic cerebral toxoplasmosis.

石榴皮中总黄酮超声辅助提取及抗氧化性分析

以石榴(Punica granatum L.)果皮为原料,采用单因素试验研究了乙醇体积分数、超声时间、料液比(m/V,g∶mL)和提取次数等因素对提取石榴皮中总黄酮的影响。结果表明,总黄酮的最适提取工艺条件为乙醇体积分数70%,超声时间20 min,料液比1∶20,提取两次。总黄酮浓度在0.20 mg/mL时清除羟自由基能力最强。

响应面法优化石榴皮中原花青素提取工艺

以原花青素提取液的吸光度(A550 nm)为指标,在单因素试验的基础上运用响应面试验考察乙醇溶液体积分数、料液比、提取时间和提取温度对石榴(Punica granatun)皮中原花青素提取率的影响。结果表明,石榴皮中原花青素的最佳提取工艺为乙醇溶液体积分数73%、料液比1∶28(m∶V,g/mL)、提取时间73min、提取温度80℃,采用高效液相色谱法测得此条件下所得石榴皮中原花青素的提取率为126.68 mg/g。

石榴花中多酚、黄酮及萜类物质同步提取工艺优化

为更有效地利用石榴（Punica granaum L.）花中多种活性成分,以石榴花为原料,利用乙醇溶剂浸提法,在单因素试验的基础上选择提取温度、提取时间、液固比、乙醇体积分数4个因素进行正交试验,优化了石榴花中3种活性物质同步提取工艺。结果表明,影响石榴花中多酚、黄酮及萜类物质同步提取的因素大小顺序为提取温度〉提取时间〉乙醇体积分数〉液固比,其最优的工艺条件为60%乙醇溶液作为提取溶剂,液固比20∶1（m L∶g）,85℃条件下提取2.0 h;在此条件下多酚、黄酮及三萜类物质的提取率分别是22.19%、6.53%、0.94%。石榴花中多酚、黄酮及萜类物质可以同步提取出来,可有效提高石榴花资源的利用率。

泰山红石榴果实生长发育动态研究

以泰山红石榴(Punica granatum L.cv.Taishanhong)为试材,于花后1周开始观测果实体积、果实直径、平均单果重、子粒百粒重、果实中总可滴定酸含量、总可溶性固形物含量等各项果实发育相关指标,寻求其动态变化规律,以全面了解石榴果实生长发育动态。结果表明,泰山红石榴果实生长发育约历时5个月,经历幼果期、果实膨大期、果实转色期和果实成熟期4个时期。随着果实体积的增加,果实出现3次迅速生长期,分别在花后4周、13周和19周。果实生长发育期内,果实体积、果实直径、平均单果重、子粒百粒重、总可滴定酸含量及总可溶性固形物含量两两均存在相关性,且均达显著以上水平,这些相关性可为生长发育期的判定提供参考。

响应面法优化超声波辅助提取河阴石榴多糖工艺研究

为了优化河阴石榴(Punica granatum L.)多糖的超声波提取工艺,研究了超声波提取时的提取温度、液料比、超声功率和超声时间单因素条件对河阴石榴多糖提取效果的影响,在此基础上,根据BoxBenhnken中心组合试验设计原理,采用三因素三水平响应面分析法建立二次回归模型,同时对各因素和因素交互作用进行方差分析,从而确定河阴石榴多糖超声提取的最佳工艺条件为提取温度48.5℃,提取时间36 min,超声波功率390 W。在此条件下,石榴多糖提取率的预期值为15.95%,试验验证值为15.68%。将优化后的试验条件与传统的水提法相比,不但提高了石榴多糖提取的效率,而且多糖得率提高了5.79个百分点。

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.

石榴3-脱氢奎尼酸合成酶基因的克隆及表达分析

利用RT-PCR技术从泰山红石榴(Punica granatum L.)果皮中获得3-脱氢奎尼酸合成酶(DHQS)基因cDNA序列,分析了其在不同石榴品种发育期果实中的表达情况。结果表明,DHQS基因cDNA序列长273 bp,编码91个氨基酸,编码蛋白具有典型的DHQ_Fe_ADH超家族保守结构域。该基因核苷酸序列与欧洲山毛榉、梅花和蒺藜苜蓿的相似性分别为91%、87%和88%,氨基酸序列与欧洲山毛榉、苹果、白梨和拟南芥的相似性分别为95%、92%、92%和88%。两个石榴品种发育期果皮中DHQS基因相对表达量逐渐降低,均在7月15日出现峰值。整个发育期,泰山红DHQS基因表达量高于泰山三白甜。

响应面法对超声波-螯合剂辅助提取石榴子多酚的优化

以石榴(Punica granatum L.)子为研究对象,以多酚得率为评价指标,通过单因素试验和响应面法分析,确定超声波-螯合剂辅助提取石榴子多酚的最佳工艺条件为乙醇体积分数50%、提取温度60℃、提取时间60 min、料液比1∶31(m:v)、螯合剂六偏磷酸钠添加量0.15%。在此条件下,石榴子多酚得率可达9.375 mg/g。通过响应面法优化得出的回归方程具有一定的实践指导意义,为石榴子资源的开发提供了参考。

不同保干促活处理对栾树和石榴移栽苗生长的影响

为加强造林质量管理,提高移栽苗的成活率和生长量,以中国常见绿化树种栾树(Koelreuteria paniculata)和经济林石榴(Punica granatum)为试验对象,采用一系列保干促活措施探索移栽后迟发苗木的挽救策略。在制定苗干发芽成活分级标准的基础上,对各处理组苗木栽后不同时期的发芽成活率、苗干成活指数、生长量、含水率及根系质量进行了调查。结果表明,处理L3和S3发芽成活率和苗干成活指数比对照组(L1和S1)分别提高了19.30、52.01个百分点和11.90、13.50个百分点,缓苗期也比L1和S1分别缩短了60和75 d。年生长量方面,L3和S3试验后平均株高和地(米)径均高于其他处理,比试验前分别增长了45.33%、57.84%和38.14%、81.82%。L3和S3的保水措施可同时有效提高苗干含水率,促进根系成活和新生根的萌发。该移栽苗保干促活挽救措施,为提升大面积生态林和经济林造林成活率及质量提供了技术参考。

石榴F3’H基因PCR反应体系的研究

从石榴(Punica granatum L.)泰山红品种的花瓣中提取总RNA并反转录得到cDNA。设计简并引物对石榴类黄酮3’-羟化酶(F3’H)基因进行PCR扩增,筛选合适的引物及退火温度。结果表明,适合石榴F3’H基因扩增的正反向引物分别为5′-CGTNGAYGTBGTBGTBGCSKCVTC-3′,5′-TCHCCDGCWATG-GCCCAHAYRTT-3′。PCR反应程序为94℃预变性5 min;94℃变性40 s,54℃退火45 s,72℃延伸60 s,共35个循环;72℃保温10 min。

不同品种石榴叶片制茶品质特性的比较

为加工优质石榴(Punica granatum L.)叶茶叶提供科学依据,以9个石榴品种的嫩叶为原料,采用绿茶传统制作工艺分别制得石榴叶茶叶;感官评定各茶样的品质,通过紫外分光光度法及高效液相色谱法对样品中的总酚、儿茶素、鞣花酸成分进行定量分析。结果显示,不同石榴品种所制茶叶品质存在较大差异,总酚、儿茶素、鞣花酸含量也各不相同,其中品种岗榴的总酚含量最高,为191.65 mg/g,大红袍含量最低,为75.53 mg/g;四川青皮的儿茶素含量最高,为7.52 mg/g,含量最低的是观赏石榴,仅为2.98 mg/g;突尼斯软子的鞣花酸含量最高,为4.76 mg/g,大红袍的含量最低,为1.88 mg/g。综合感官评定与化学成分分析结果,石榴品种三白的叶片制茶品质最佳,其次是枣庄青皮、四川青皮和甜绿子。

第一届中国石榴博览会暨第六届全国石榴生产与科研研讨会在河南荥阳举行

第一届中国石榴博览会暨第六届全国石榴生产与科研研讨会于2015年9月19日至9月21日在河南省荥阳市顺利召开。本届大会以“互联网＋石榴”为主题,由中国园艺学会石榴分会和中国农业科学院郑州果树研究所主办,郑州市休闲观光农业协会、荥阳市河阴石榴产业基地承办,河南农业大学协办。开幕式由中国农业科学院郑州果树研究所所长刘君璞主持。荥阳市政协副主席刘阳、中国园艺学会名誉理事长朱德蔚、中国经济林协会常务副会长兼秘书长伍步生分别致辞。

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.

石榴果皮粗提物的生物活性初步研究

利用70％（v／v）甲醇浸提石榴（Punica granatum L．）果皮，分别用石油醚、氯仿、乙酸乙酯、正丁醇萃取，得到4种萃取物漫膏。试验了4种萃取物浸膏对黄瓜和萝卜种子发芽和生长的影响。结果表明，除高浓度石油醚萃取物浸膏对黄瓜种子发芽有抑制作用外，其他萃取物浸膏对黄瓜种子发芽表现出促进作用；氯仿萃取物浸膏对萝卜发芽抑制作用较强；乙酸乙酯和正丁醇萃取物浸膏对黄瓜根长表现出促进作用，但根重的促进作用不明显；其他萃取物浸膏对种芽的生长和根的生长总体上表现为抑制作用。

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.

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.

石榴低温响应因子CBF基因家族鉴定及其表达分析

CBF(C-repeat binding factor)是AP2家族的一类转录激活因子,在植物响应低温胁迫和提高植物耐寒性方面具有重要作用。虽然多个石榴品种的基因组已经发布,但仍缺乏对其CBF基因家族的全面研究。为全面分析石榴(Punica granatum L.)CBF基因家族分子生物学特性,本文对鉴定到的PgCBFs成员进行了生物信息和表达分析。结果表明,石榴全基因组中有7个CBF基因家族成员,蛋白序列中除包含AP2结构域外,还具有CBF特征序列PKKPAGRxKFxETRHP和DSAWR;7个成员集中分布在染色体1和4上,编码蛋白质氨基酸长度为201~267aa,分子质量为21.69~29.81ku;进化分析显示PgCBFs基因家族成员分布在GroupⅡ~Ⅳ亚组中,与水稻CBF成员组成的GroupⅠ存在明显区分;除PgCBF2、PgCBF4、PgCBF5基因结构中含有1~2个内含子外,其余与其他物种类似,属内含子缺失型;编码的蛋白序列均含有保守基序Motif1~7;蛋白二级结构主要为不规则卷曲和α-螺旋,三级结构较为相似。PgCBFs基因家族扩增主要来源于串联重复,并与拟南芥、苹果、桃分别存在2对、5对和4对共线性关系;GO注释显示PgCBFs多与转录调控、低温胁迫或激素诱导相关;启动子区含有多种顺势作用元件,主要为胁迫刺激、激素诱导和光响应元件;基因表达分析发现除PgCBF6外,其余成员在根中高度表达,在低温胁迫处理下PgCBF在根和韧皮部表达量显著上调,其中PgCBF7能够快速响应并持续应答低温诱导。结合进化树、共线性、启动子以及表达分析,推测PgCBF7可能与石榴幼苗低温胁迫调控相关。本研究可为深入研究石榴CBF基因家族生物学功能提供理论基础。

石榴在南水北调中线核心水源区的引种试验报告

十堰市具有独特的自然生态环境,尤其是南水北调工程核心水源区的小气候条件特殊。为探索适宜该区域农业结构调整发展的种植新项目和适应水源区生态保护的种植新模式,培育本土化的优良石榴(Punica granatum L.)品种,从2009年开始,水源区由河南省荥阳市引进突尼斯软子石榴进行试种,结果综合表现明显超出了原引种地,其中优系的早实、丰产性好,果实经济性状优、商品性好,花果美艳、观赏价值高,生态经济效益十分可观,值得大力推广发展。