不同干燥方式对无核紫葡萄干品质的影响

采用自然干燥和太阳能干燥2种方式,对无核紫葡萄进行干燥研究,每种干燥方式又分别采用无处理和促干剂浸泡处理。测定在2种干燥方式下所得无核紫葡萄干的水分、总糖、总酸、矿物质元素、花青素和沙尘含量。结果表明,无核紫葡萄干水分含量为15%～19%;总糖含量约74%;总酸含量为1.4%～1.7%;VC含量采用太阳能干燥的比自然干燥的高;花青素含量是经促干剂处理的比未经促干剂处理的含量高,而经太阳能干燥的比经自然干燥的含量低;太阳能干燥比自然干燥的葡萄干中含沙尘量低。     

气体射流冲击干燥无核紫葡萄的数学模型

针对气体射流冲击干燥技术,试验研究了不同风温、风速和预处理下无核紫葡萄的4种薄层干燥模型。在对无核紫葡萄进行L9(34)正交干燥试验的基础上,通过试验数据分析,对不同风温下的干燥曲线进行模型比较,经拟合得出适合无核紫的数学模型为Page模型。     

农产品中白藜芦醇的高效液相色谱标准化测定方法研究

旨在建立同时测定花生衣、葡萄皮、苜蓿、虎杖、藜芦等农产品中白藜芦醇含量的高效液相色谱标准化测定方法。色谱分析条件如下:以Waters Atlantis T3-C18色谱柱(4.6×150mm,3μm)分离,柱温40℃;以乙腈:0.1%乙酸溶液(体积比为26:74)为流动相,流速1.0 mL/min;采用二极管阵列检测器,检测波长306 nm。花生衣、葡萄皮、苜蓿中白藜芦醇含量的检出限均为0.6μg/g,定量限均为2.0μg/g;虎杖、藜芦中白藜芦醇含量的检出限均为7.5μg/g,定量限均为25μg/g。方法学考察结果显示符合含量测定要求。该方法操作简便、快速,结果可靠,重现性好,可用来研究农产品中白藜芦醇的含量。     

RP-HPLC测定干姜中6-姜酚的含量

采用RP-HPLC法,使用依利特ODS色谱柱(Hypersil,4.6 mm×150 mm,5μm),以乙腈—0.1%醋酸水溶液(55∶45)为流动相,流速1.0 mL/min,检测波长280 nm,柱温35℃,检测干姜中6-姜酚的含量。结果表明,标准曲线线性相关系数R为0.999 8;连续进样5次,测定的峰面积RSD为0.47%;同一样品3次检测结果的最大相对偏差为4.4%;适合于作为干姜中6-姜酚含量的测定方法。     

高速逆流色谱纯化白藜芦醇的研究

利用高速逆流色谱分离纯化虎杖中的白藜芦醇.实验结果表明,选择溶剂体系为,氯仿:甲醇:水=4:3:2,均为转速850 r/min,流速2 mL/min,紫外检测波长280 nm.所得产品经高效液相色谱测定,白藜芦醇得率为0.186%,纯度大于96%.     

高速逆流色谱纯化白藜芦醇的研究

利用高速逆流色谱分离纯化虎杖中的白藜芦醇.实验结果表明,选择溶剂体系为,氯仿：甲醇：水=4：3：2,均为转速850 r/min,流速2 mL/min,紫外检测波长280 nm.所得产品经高效液相色谱测定,白藜芦醇得率为0.186%,纯度大于96%.     

鸡蛋中唾液酸含量的测定

采用高效液相色谱法检测鸡蛋中唾液酸的含量。鸡蛋中的唾液酸在浓度0.5mol/L的NaHSO4溶液中水解30min后,以邻苯二胺为衍生化试剂,在温度80℃水浴中衍生40min。采用AlltimaC18柱(250mm×4.6mm,5μm),流动相为1.0%四氢呋喃水溶液(含0.2%磷酸)-乙腈(92∶8),流速为1.0mL/min,柱温为35℃,紫外检测波长230nm。此方法具有灵敏度高、重复性好等特点,可以准确地测定鸡蛋中的唾液酸含量和蛋制品中鸡蛋的含量。     

利用SCAR标记SCF27对葡萄无核性状的鉴定研究

本研究以14份葡萄种质资源和火州黑玉葡萄3个杂交组合共116株胚挽救后代为试材,研究葡萄无核性状的鉴定方法,以期在苗期实现杂交单株无核性状鉴定,提高无核葡萄育种效率。利用SCAR标记SCF27对材料进行无核性状分子检测,通过田间鉴定对检测结果进行验证和评价。结果表明,SCF27标记在‘无核白’、‘火州黑玉’、‘火焰无核’、‘红宝石无核’、‘美丽无核’,‘火州红玉’、‘火州紫玉’、‘无核紫’、‘无核白鸡心’、‘昆香无核’、‘底来特’等11个无核品种植株中均检测到目的无核性状基因片段,而在‘红地球’、‘贵妃玫瑰’和‘早艳’等3个有核品种植株中均未检测到,在116株后代中检测出84株携带目的基因片段,其中81株与田间鉴定结果吻合,吻合率为96.4%。本研究认为,SCF27标记是一个显性或主效基因标记,其检测结果与植株表现型的吻合率很高,可以用来进行葡萄杂交后代无核单株的早期筛选,但鉴定结果可能存在一定误差。     

高效液相色谱法快速测定淫羊藿中6种主要黄酮成分

建立了RP-HPLC法快速测定淫羊藿中6种主要黄酮成分的方法。色谱条件为:色谱柱:WelchromTMC18(200mm×4.6mm,5μm);流动相:乙腈—水,梯度洗脱;柱温:35℃;流速:1mL/min;检测波长:270nm。结果表明,6种黄酮在27min内达到基线分离。该方法分析时间短、稳定性和准确度良好,对淫羊藿药材的质量控制具有一定的参考价值。     

免疫亲和柱净化高效液相色谱法测定坚果中黄曲霉毒素

建立了免疫亲和柱净化高效液相色谱法测定坚果中黄曲霉毒素B1,B2,G1和G2总量。样品用甲醇—水提取,经免疫亲和色谱柱纯化,应用液相色谱法检测。试验结果表明:空白样品分别按照5.2,26,52μg/kg添加黄曲霉毒素,回收率为81.3%～96.0%,精密度<10%,黄曲霉毒素B1,B2,G1和G2的检测灵敏度分别为0.10,0.05,0.10,0.15μg/kg。免疫亲和柱净化高效液相色谱法测定坚果中黄曲霉毒素B1,B2,G1,G2总量,是一种简单、快速和准确的方法。     

大孔吸附树脂对虎杖中白藜芦醇的分离纯化研究

旨在筛选出分离纯化虎杖中白藜芦醇的最佳大孔吸附树脂以及工艺条件。静态吸附与解吸、动态吸附与解吸通相结合的方法,以树脂的最大吸附量、解析率为考察指标,确定最佳的纯化工艺。H103树脂对白藜芦醇有较好的吸附与解吸效果,其最佳的工艺条件为粗提液中白藜芦醇的质量浓度为0.72 mg/mL、上样流速2 BV/h,吸附饱和后先用4 BV的蒸馏水进行洗涤,然后用8 BV、75%的乙醇溶液以1.5 BV/h的流速进行洗脱,白藜芦醇的含量可由纯化前的12.8%提升至53.5%。应用H103树脂对虎杖中的白藜芦醇进行纯化,其工艺稳定可行,具有吸附快、解吸容易、解吸液安全低毒且回收容易,具有较高的应用价值。     

Improving in vitro induction of autopolyploidy in grapevine seedless cultivars

The efficiency of in vitro polyploidization depends on several variables associated to the plant, the antimicrotubule agent and the interactions between them. In the present work, we have used response-surface methodology to determine the best operating conditions for plant recovery in polyploidization assays for shoot apices and somatic embryos of two seedless grape cultivars, employing colchicine and oryzalin. Explant type, tubulin-interfering compound and concentration were the critical factors determining plant recovery. Linear reduction in viable plant regeneration via organogenesis and somatic embryogenesis was obtained by increasing oryzalin concentrations and treatment time, whereas the effects of colchicine were better described by a quadratic design for both explants types. The conditions promoting higher rates of plant recovery were used in chromosome doubling experiments with oryzalin and colchicine for shoot apices and somatic embryos of ‘Crimson seedless’ and ‘BRS Clara’. The established protocols allowed the recovery of non-chimerical autotetraploid plants at rates higher than 30 % for both cultivars. Stomata size parameters statistically correlate to the ploidy level of the regenerants and were effective for preliminary polyploidy screening. Autotetraploid lines of seedless grapes were incorporated into the Vitis germplasm bank for further agronomical evaluations. To our knowledge, this is the first report of in vitro oryzalin induced polyploidization of grapevine and of the use of mathematical modeling to optimize chromosome doubling in plants.     

静态顶空气相色谱法快速测定葡萄中乙醇和乙醛含量的研究

介绍了建立静态顶空气相色谱法测定贮藏后玫瑰香葡萄中乙醇、乙醛含量的方法。对顶空进样条件进行了优化,顶空温度65℃,恒温加热时间为35min最优。此方法的乙醇、乙醛检测限分别达到0.02mg/kg和0.03mg/kg,其回收率分别为98.7%和95.6%,与其它测定方法相比具有操作简单,分析快速,结果准确等优点。     

A study on quality loss of minimally processed grapes as affected by film packaging

The influence of film barrier properties on the quality loss of minimally processed grapes stored at 5 °C was addressed. Table grapes (Vitis vinifera cv. Italia) differing in quantity and frequency of irrigation, were tested with five different packaging films. Two commercially available films were used: a multilayer film obtained by laminating nylon and a polyolefin layer (NP), an oriented polypropylene film (OPP), along with three biodegradable polyester-based films (NVT-100, NVT-50 and NVT-35). The packed grape quality during storage was determined by monitoring the headspace oxygen and carbon dioxide concentration, the grape sensory qualities, and the viable cell concentration of the following spoilage microorganisms: total viable bacterial count, lactic acid bacteria, yeasts and molds. All the investigated films successfully preserved the quality of packed produce for the entire observation period (35 d). However, the best results were obtained using high barrier films such as NP and NVT-100. Slight differences were recorded between the two sets of table grapes in terms of respiratory activity and sensory quality.     

Rachis browning in four table grape cultivars as affected by growth regulators or packaging

Plant growth regulators (PGRs) are used to increase berry size in table grapes. The objective in this study was to determine if the levels of PGRs used commercially affected rachis postharvest quality. Three seedless table grape cultivars, ‘Mystery’, ‘Superior’, and ‘Crimson’ were treated with gibberellin (GA), cytokinin (CPPU), or both at berry diameter of 6–8 mm, while a seeded cultivar, ‘Redglobe’ was treated at 13 mm. The fruits were harvested at commercial maturity and held for 7 days shelf-life either immediately after harvest or after 2 weeks storage at 0 °C. The combination of GA and CPPU increased berry weight and diameter, and rachis diameter in all three seedless cultivars, but had minor or no effect on TSS and acidity. Rachis quality was measured by a browning scale derived from image analysis. Browning was lower after storage for rachis of ‘Mystery’ treated with GA, while the other three cultivars showed no difference between PGR treated and control rachis. Reduction of water loss by covering punnets of control clusters with microperforated film had a significant positive effect on decreasing weight loss in all cases and a positive effect on slowing rachis browning. Calculation of the shelf-life required to reach 50% rachis browning found that shelf-life was longer for the microperforated packaging as compared to open packages in all the cultivars, with stronger influence during shelf-life after storage than shelf-life after harvest. However, the data also indicated that rachis browning cannot only be attributed to weight loss, either from the whole cluster or from the rachis.     

无核葡萄SRAP-PCR反应体系的建立和优化

以大粒无核葡萄‘皇家秋天’的基因组DNA为模板,对无核葡萄SRAP-PCR反应体系的主要成分及反应退火温度进行优化。获得的最优SRAP-PCR反应程序为94℃预变性5 min,94℃变性1 min,38℃复性1 min,72℃延伸1 min,5个循环;94℃变性1 min,56℃复性1 min,72℃延伸1 min,35个循环;72℃终延伸10 min。25 μL体系中,模板DNA 40 ng,Mg2+ 1.5 mmol/L,dNTPs 0.2 mmol/L,引物0.2 μmol/L,Taq DNA聚合酶0.6 U。实验结果表明,优化后的SRAP-PCR反应体系扩增多态性高,带型清晰,稳定性好。     

无核葡萄种质亲缘关系的SRAP研究

为充分了解及更合理地利用无核葡萄的种质资源,辅助无核葡萄育种,本研究采用SRAP分子标记技术,对23 个国内外主流无核葡萄品种的种质亲缘关系进行分析。结果表明：从45 对SRAP引物中筛选出38 对多态性引物,共扩增出236 条谱带,其中171 条为多态性谱带,多态性谱带百分率为72.4%。遗传相似分析显示,供试材料间遗传相似系数的变化范围为0.58~0.90,遗传距离在1.0313~4.9643 范围内变化。在遗传相似系数为0.61 处,可将供试材料分为2 个大类群,在相似系数为0.66 处,Ⅰ类群和Ⅱ类群均可分为2个亚群。说明23个品种总体来讲亲缘关系较远,特别欧美杂种具有较高的遗传多样性,部分欧亚种品种间亲缘关系较近,杂交育种时应注意亲本选择。     

无籽罗汉果组培无菌系的构建

【研究目的】研究无籽罗汉果组培快繁技术要点,为无籽罗汉果再生体系建立和工厂化生产提供技术支持。【方法】以幼嫩茎段为试验材料,探讨不同灭菌时间,不同激素组合以及浓度对启动培养、增殖培养、生根培养等的影响。【结果】无籽罗汉果带芽茎段最佳消毒方法为1% HgCl2溶液消毒8 min。最适宜的无籽罗汉果茎段启动培养基为MS+1.0 mg/L 6-BA+1.5 mg/L KT,无籽罗汉果继代增殖培养基为MS+1.0 mg/L 6-BA+0.5 mg/L IBA,增殖系数可达6.8;较适合生根的培养基为1/2MS+1.0 mg/L IBA+0.5 mg/LNAA,新稍平均生根数达7.4条,生根率可达95%。【结论】可为无籽罗汉果大规模组织培养和快速繁殖提供理论基础,并为组织培养高效体系的建立及外源基因导入提供参考。     

胚挽救技术创造无核抗病葡萄新种质

为提高无核抗病、优质葡萄新种质的选育效率,以无核欧洲葡萄品种作母本,中国野生刺葡萄‘塘尾’、‘雪峰’、山葡萄‘双优’及欧山杂交品种‘北醇’做父本,利用胚挽救技术对取样时间和培养基进行优化;用分子标记筛选后代的无核与抗病性状。结果表明：（1）15个杂交组合中共获得杂交果实13341个,接种幼胚珠8589枚,幼胚发育胚1338枚,平均胚发育率15.59%,正常成苗234株,平均成苗率2.73%,‘红宝石无核’ב塘尾’组合胚挽救效果最好,胚发育率为40.17%、成苗率7.76%和成苗植株数178株。（2）‘火焰无核’×北醇’授粉后42天取样,胚发育率(11.37%)和萌发率(3.52%)为最高。‘京可晶’ב北醇’授粉后38天取样,胚发育率(4.3%)和萌发率(1.08%)为最高。（3）以MM3为基础培养基,外源添加3 mmol/L的腐胺,‘红宝石无核’ב塘尾’获得胚发育率(55.83%)和成苗率(20%)为最高。（4）4个自然授粉的品种（‘昆香无核’、‘美丽无核’、‘红宝石无核’、‘赫什无核’）,其胚珠在E20A培养基中更有利于幼胚发育率提升。（5）使用葡萄无核标记SCF27-2000和GSLP1-569、抗霜霉病标记S294-369和S382-615及抗白粉病标记OPY13-661进行筛选,杂种后代中有16株同时携带无核且抗霜霉病与抗白粉病特异性条带。该试验为无核葡萄抗病育种提供了新材料,将为进一步开展中国野生葡萄育种相关研究奠定基础。