Cultivar identification using 18S–28S rDNA intergenic spacer-RFLP in pomegranate (Punica granatum L.)

Since pomegranate is a fruit tree species showing high plant diversity, molecular techniques are required to quickly and precisely characterize and certify different cultivated varieties. The study evaluates a genetic method to identify pomegranate cultivars. The procedure is based on the Restriction Fragment Length Polymorphisms (RFLP) and Polymerase Chain Reaction (PCR) techniques. Ten pomegranate accessions from the varietal groups Mollar de Elche, Mollar de Albatera, Mollar de Orihuela, Valencianas and Bordes were evaluated. The results prove the appropriateness of the PCR–RFLP technique for identifying pomegranate cultivars. All evaluated cultivars were differentiated according to their genetic profiles. There is a low correlation between pomegranate morphological and genetic traits on this study.     

Effects of foliar application of calcium chloride and urea on quantitative and qualitative characteristics of pomegranate fruits

The effects of urea and calcium chloride and their combinations on qualitative and quantitative characteristics of pomegranate fruits were studied over 2 years. Aqueous solutions of urea at 0%, 0.5%, 1% and 2% and calcium chloride at 0%, 2% and 4% were applied on two branches of pomegranate (Punica granatum L. cv Malase-Yazdi) at full bloom (FB) and 1 month after full bloom (1MAFB) stages. Urea at concentrations of 1% and 2% significantly increased aril size, fruit length and diameter. Calcium chloride at concentrations of 2% and 4% significantly increased average fruit weight and ascorbic acid (AA) content. Both urea and calcium chloride increased soluble solid content (SSC) at all of the used concentrations. In the first time of spraying, higher level of treatments reduced titratable acidity (TA), but the second time of spraying did not significantly affect the TA.     

Evaluation of genetic diversity among Iranian soft-seed pomegranate accessions by fruit characteristics and RAPD markers

Soft-seedness in pomegranate is a desirable trait for fresh consumption of this valuable fruit. At the main Iran pomegranate collection, 21 pomegranate accessions gathered from different parts of Iran are registered as soft-seed genotypes. The aim of this research was to study these soft-seed pomegranate accessions using fruit morphopomological traits and DNA markers to reveal their relatedness. Thirty-six fruit characteristics were measured in these accessions together with applying 29 random decamer primers already reported to be polymorphic on pomegranate. Factor analysis on mean values of fruit characteristics determined 10 main factors and applied for grouping of the accessions using Ward's method. Also 14 of the random primers showed good amplification and polymorphism on these samples, and a total of 43 RAPD markers were produced. Estimates of genetic similarity, using Jaccard's similarity coefficient, ranged from 0.13 to 1.0 using the RAPD data. Grouping based on the fruit traits compared with that based on RAPD data did not produce a significant correlation (r = −0.36). Morphometric measurements and sensory evaluation confirmed that some accessions are hard or semi-hard seeded. This study showed that information based on fruit characteristics and RAPD markers are complementary for genetic discrimination in soft-seed pomegranate accessions. This might be due to the high level of similarity between soft-seed pomegranate accessions.     

石榴不同枝梢完全花比例及坐果率调查

以会理青皮软籽石榴为试验材料,研究石榴不同枝梢完全花比例及坐果率。结果表明：①不同枝梢完全花占开花量的比例：极短枝（33.49%）＞短枝(21.51%)＞中枝(8.81%)＞长枝(7.32%);②不同枝梢坐果率：极短枝(6.60%）＞短枝(5.28%）＞中枝(1.55%）＞长枝(0.24%）。结论：①极短枝、短枝是石榴坐果的关键枝;②石榴头茬花的完全花比例高、坐果率高。     

4种改良CTAB法提取石榴成熟叶片DNA的比较研究

【目的】研究从成熟石榴叶片中提取高质量基因组DNA的方法。【方法】根据成熟石榴叶片组织中富含多酚、多糖的特点,以峄城中国石榴种质资源圃20种石榴成熟叶片为材料,分别采用改良CTAB法(Ⅰ、Ⅱ、Ⅲ、Ⅳ)4种方法提取成熟石榴基因组DNA,通过琼脂糖凝胶、紫外分光光度计分析、ISSR扩增等方法检测所提取DNA的质量。【结果】改良的CTAB Ⅳ法能从各个石榴品种的成熟叶片中提取出基因组DNA,琼脂糖电泳胶孔干净,无拖带,且DNA的纯度和完整性都较好,OD260 nm/OD280 nm的比值均在1.8~2.0之间,无降解现象,其质量、产量都高于其他改良CTAB法。经ISSR- PCR扩增结果表明,此方法提取的基因组总DNA完全适于ISSR分析。其主要的步骤是：在石榴叶片的研磨中添加Vc、PVP等抗氧化物质,加入CTAB提取液后65 ℃水浴30 min后冷却至室温,离心条件为15℃,10 000 r/min,10 min,DNA粗提液用氯仿抽提3次,可获得高质量的DNA。【结论】改良的CTAB Ⅳ法能有效地从石榴成熟叶片中提取到高质量的基因组DNA。     

不同石榴品种果实不同部位酚类物质含量和抗氧化能力比较

为分析不同石榴品种果实不同部位的酚类特性,以枣庄地区13个石榴品种成熟期果实为材料,利用比色法测定果皮、果汁和种子总酚、单宁、总类黄酮、原花色素、黄烷醇等的含量,采用DPPH清除法分析抗氧化能力。结果表明,各石榴品种间且同一品种不同部位的酚类物质存在明显差异,但均以果皮中酚类物质含量最高。‘软籽谢花甜’果皮总酚、单宁、总类黄酮含量均最高,种子总酚、原花色素、黄烷醇含量也最高,而‘小青皮酸’果汁中总酚、单宁含量最高。各石榴品种果皮、种子抗氧化能力和总酚、单宁、总类黄酮含量之间均为显著正相关,含量高的品种抗氧化能力也强。主成分分析将果皮、种子和果汁中5~6个酚类物质指标简化为2个主成分值来衡量酚类物质含量,不同石榴品种不同部位的主成分值不同。主成分值排在前5名的‘软籽谢花甜’是值得大力推广和深入开发利用的优良品种。可对酚类物质含量较高的石榴皮进行综合利用,开发相关产品。     

石榴皮和石榴渣总黄酮含量的测定

黄酮类化合物是自然界广泛分布于植物中的一大类天然产物,其结构类型的多样性和生物活性的光谱性,已成为国内外研究开发的热点。目前多以芦丁、槲皮素为标准品,进行植物中黄酮含量的测定。本试验以芦丁为标准品,采用分光光度法,对南疆石榴皮、石榴渣中总黄酮的含量进行了测定。结果表明,石榴皮中总黄酮含量为6.65%,石榴渣中总黄酮含量为2.04%。这对南疆石榴资源的合理利用提供了参考依据。     

云南蒙自石榴叶霉病病原菌鉴定

[目的]鉴定云南蒙自石榴（Phomopsis punicae Linn.）霉病病原菌,为进一步研究蒙自石榴叶霉病有效防治方法奠定基础。[方法]病原分离采用组织分离法,通过分生孢子悬浮液接种,用柯赫氏法则对其致病性进行验证。显微镜下观察菌丝形态及产孢结构,测量孢子大小,根据形态特征进行鉴定。[结果]显微镜下,分生孢子梗单生,直立,顶部微弯或曲膝状,深褐色,有隔膜,（129.3～352.5）μm×（3.5～4.7）μm;枝孢短链生,褐色,有或无隔膜,具油滴,（9.0～36.0）μm×（4.0～5.0）μm;分生孢子链生,椭圆形,近球形,褐色,无隔膜,具油滴,（4.0～8.0）μm×（3.0～4.0）μm。[结论]叶霉病发生在石榴生长后期,一般危害石榴老叶,形成近圆形或不规则形黑褐色病斑,病斑上长橄榄绿色霉层,为病原菌的子实体。造成石榴提早落叶。其病原菌鉴定为极细枝孢（Cladosporium tenuissimum Cooke）。     

石榴皮有效抑菌部位萃取工艺的比较分析

[目的]优化石榴皮有效抑菌部位的萃取工艺。[方法]采用超声辅助萃取法,选取纯水、甲醇、乙醇、丙酮、乙酸乙酯、正己烷6种溶剂萃取石榴皮抑菌药液。通过琼脂平板法测定石榴皮抑菌药液对9株水产常见致病菌的最低抑菌浓度,正交试验方法考察溶剂浓度、料液比、温度、超声时间对抑菌率的影响,进一步优化石榴皮抑菌药液的萃取工艺。[结果]比较不同溶剂抑菌效果,表明乙醇为最佳萃取溶剂,其萃取部位平均最低抑菌浓度为0.517 mg/m L。在乙醇体积分数70%、料液比1∶20 g/m L、萃取温度60℃、超声时间40 min的最佳萃取工艺条件下,萃取药液平均最低抑菌浓度为0.442 mg/m L。[结论]研究可为石榴皮在水产养殖细菌性疾病防控中的应用和石榴皮抗菌剂的研制及工业化生产提供理论依据。     

怀远石榴生长的气候与土壤条件分析

地处亚热带至暖温带过渡的怀远县，兼有南北气候特征的生物气候带为石榴生长提供了充裕的光、温、水资源；麻石棕壤上生长的石榴产量高、品质优。本文分析了怀远石榴产量、品质与气候、土壤条件的关系，为石榴的因土种植提供了科学依据。