乙醇沉淀法提取石榴皮中多糖类研究

以石榴皮为原料，在进行单因素试验的基础上，对料液比、沉淀时间做了均匀设计试验，得到乙醇沉淀法提取石榴皮中多糖类化合物的非线性回归方程为：y=1.650—0．295x1-0．006x2＋0．103x1^2，进而获得最佳提取条件为：料液比1：5，沉淀时间2h，离心时间10min。     

红花石榴二氢黄酮醇还原酶(DFR)基因cDNA片段克隆及序列分析

二氢黄酮醇还原酶(DFR)是植物花色苷生物合成途径中的关键酶。以红花石榴"泰山红"的花瓣总RNA为模板,利用简并引物,采用RT-PCR方法,扩增出一条长446 bp的cDNA片段,序列分析结果表明,该cDNA片段与GenBank中的甜樱桃、葡萄等物种的DFR氨基酸同源性为78%～81%,说明该二氢黄酮醇还原酶片段为石榴花DFR基因的cDNA片段,GenBank登录号为JN316028。     

Effect of feeding pomegranate seed oil as a source of conjugated linolenic acid on Arabian stallion semen quality in cooled and postthawed condition

The objective was to assess the influence of pomegranate seed oil supplementation on the quality of fresh, cooled and frozen–thawed Arabian breed stallion semen. Eight stallions (n = 4 per group) received their normal diet (control group) or normal diet top dressed with 200 ml of pomegranate seed oil (PSO group). Semen was collected every fifteen days for 90 days. Stallions were reversed across the treatments after a sixty‐day interval. In cooled and stored condition (2, 12 and 24 hr), spermatozoa motion characteristics, membrane integrity, viability, morphology and lipid peroxidation were analysed. In frozen–thawed semen, sperm dynamic characteristics were analysed by CASA, acrosome status and mitochondrial activity (evaluated by Flow cytometry) determined. The effects of treatment, time, semen type and their interactions were submitted to PROCMIX (SAS^®), and means compared by the Tukey test. Also, collected semen samples were artificially inseminated to evaluate fertility and pregnancy rate after day 60 of the experiment. The results from fresh condition showed that semen volume, sperm concentration, abnormality and live sperm were not affected by dietary treatment (p > 0.05). In cooled condition, the higher value for sperm plasma membrane integrity and viability was observed in PSO group compared to control after 24 hr cooled and stored in 5°C. In postthawed condition, the higher value for CASA total motility and acrosome status was observed in PSO group compared to control group (p < 0.05). One hundred and twenty‐six mares were artificially inseminated for fertility trial using control and PSO groups’ fresh semen. The average pregnancy rates were not significantly different between control and treated group (62.88% and 65.90%, respectively) (p > 0.05). We concluded that under the conditions of this study, dietary supplementation of 200 ml pomegranate seed oil seems to relatively improved Arabian horse sperm quality during storage in cooled and frozen condition via increasing plasma membrane integrity, viability and acrosome status, but did not improve the pregnancy rates.     

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。     

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。     

不同激素和黑暗处理对泰山红石榴离体叶片再生的影响

以泰山红石榴为试材,研究了不同激素配比、黑暗处理时间等关键因素对泰山红石榴离体叶片愈伤组织诱导和不定芽再生的影响。结果表明,细胞分裂素是离体叶片再生不定芽所必需的。生长素2,4-D与IBA组合诱导出的愈伤组织生长量大,但不定芽再生率低。黑暗处理时间以2周为宜。适宜的再生培养基为MS+KT1.0mg·L-1+BA2.0mg·L-1+GA30.5mg·L-1,不定芽再生率达57.0%,每片叶再生芽数为3.0个。     

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.     

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。