柑桔溃疡病菌的PCR检测方法研究

通过优化PCR反应条件,拟建立快速、特异、准确的检测柑桔溃疡病的方法。试验结果表明,25μL优化反应体系:10×PCR缓冲液2.5μL,10 mM/L dNTPs 0.5μL(终浓度为0.2 mM/L),2 U/μLTaq DNA聚合酶0.5μL(终浓度为0.04 U/μL),5 mM/L引物各0.5μL(终浓度为0.1 mM/L),DNA模板1μL(终浓度为6.0×10~6cfu/mL/mL),灭菌ddH_20 19.5μL;PCR扩增退火温度为59℃;柑桔溃疡病菌病茵浓度检测下限为6.0×10~3cfu/mL,可以获得柑桔溃疡病菌的特异性片段,能够满足生产中对柑桔溃疡病检测的要求。     

花椰菜最适ISSR反应体系的建立及优化

以花椰菜9901A×9908杂交种为材料,用高盐低pH值法提取的基因组DNA为模板,对ISSR-PCR反应体系中的镁离子浓度、dNTP浓度、Taq DNA聚合酶量、引物用量以及最适退火温度等影响因子进行了优化和筛选,建立了适合花椰菜的ISSR反应体系:20 μL PCR反应体系,含10×PCR反应缓冲液2 μL,2.0 mmol/L MgCl2,4×dNTP混合物0.2 mmol/L,引物0.5 μmol/L,Taq酶1.5 U,基因组DNA约30 ng,最适退火温度为52.8℃.     

利用正交设计建立洋桔梗RAPD最佳反应体系

利用正交设计法对影响洋桔梗RAPD反应的Mg2 、dNTPs、Taq DNA 聚合酶、引物和模板等主要因素的作用进行了比较分析,建立了洋桔梗RAPD的最佳反应体系:即在25 μL反应体积中,10×PCR buffer为2.5 μL ,Mg2 浓度为2.0 mmol/L,dNTPs浓度为0.4 mmol/L,引物浓度为0.2 μmol/L,模板浓度为50 ng/μL,Taq DNA聚合酶用量为1.0 U;在此基础上,比较了不同退火温度对RAPD扩增产物的影响,退火温度经筛选选定为41℃;最后再将该反应体系应用于不同引物,结果表明该优化结果具有较好的稳定性和通用性.     

百合三种病毒的多重RT-PCR检测

根据基因库中黄瓜花叶病毒（Cucumber mosaic virus,CMV）、百合斑驳病毒（Lily mottle virus,LMoV）、百合无症病毒（Lily symptomless virus, LSV）的外壳蛋白基因序列,设计了3对特异引物,通过对扩增条件进行优化,建立了同时检测CMV、LMoV和LSV的多重RT-PCR检测方法。该方法可以从带有CMV、LMoV和LSV的样品中同时扩增出3条大小与试验设计相符的657 bp (CMV) 、428 bp（LMoV）、171 bp (LSV)的特异性多重RT-PCR扩增带。扩增产物测序表明,CMV、LMoV和LSV 3种病毒与GenBank中登录的亚洲和荷兰多数分离物核苷酸同源性在90%以上,地域差异不明显,外壳蛋白序列高度保守。     

山梨SSR反应体系的建立及其稳定性验证

以山梨为试材,提取基因组DNA,运用5因素5水平正交实验设计,对SSR反应体系中的DNA模板、Mg2+、Taq酶、dNTPs和引物用量进行优化试验,确立适宜的SSR反应体系。结果表明:20μL反应体系中,模板DNA 10ng、Mg2+(20mmol/L)2.5μL、Taq酶1.25U、dNTPs(10mmol/L)1.0μL、引物(10μmol/L)1.5μL、10×PCR buffer 2.0μL;应用该SSR体系,在引物筛选试验及山梨×"幸水"后代群体中进行扩增,扩增效果较好,证实了该体系的适用性和稳定性。     

利用正交设计优化辣椒SRAP反应体系

为快速建立优化的辣椒SRAP反应体系,利用L9(34)正交表,探讨了Mg2+、dNTPs、引物和模板DNA4种主要反应成分的浓度变化对SRAP扩增结果的影响。结果表明:通过正交设计实验可高效建立优化稳定的辣椒SRAP反应体系;用该法建立的辣椒SRAP优化反应体系为:20μL反应体系中含10×PCRBuffer 2.0μL、Mg2+2.5 mmol/L、dNTPs 0.2 mmol/L,上下游引物各0.4μmol/L、模板DNA10 ng,TaqDNA聚合酶1 U。     

苹果茎沟病毒实时荧光RT-PCR反应体系的优化

以健康昆诺阿藜叶片和受ASGV侵染的昆诺阿藜叶片为试材,利用Trizol试剂提取植物总RNA,将实时荧光RT-PCR体系的主要成分设定梯度,根据每个成分的变化引起的(Rn值和Ct值的差异,探讨苹果茎沟病毒实时荧光RT-PCR技术中反应体系的主要成分对扩增结果的影响.最终确定了RT-PCR反应体系的最佳条件为:25 μL体系中,Mg2+浓度为3.5 mM,引物浓度为0.6 μM,探针浓度为0.6 μM,总RNA含量为20 ng.利用此反应体系进行扩增,△Rn值比未优化时高出一个数量级,表明该体系适合ASGV实时荧光RT-PCR检测分析.     

枇杷iPBS分子标记的PCR体系优化与引物筛选

以枇杷的DNA为模板进行iPBS-PCR扩增,采用预备试验中效果较好的iPBS分子标记引物2241,对枇杷iPBS-PCR反应体系中的dNTP、Mg2+、引物、模板用量进行优化。根据检测结果确定枇杷iPBS-PCR使用10×Taq buffer 2μL,25mM Mg2+1.6μL,2.5mM dNTP 1.6μL,10μmol/L Primer 1μL,5U/μL Taq酶0.2μL,模板DNA 5ng的20μL反应体系。反应程序中的退火温度可以参考iPBS引物理论Tm值。对33条iPBS引物扩增测试的结果显示在该反应体系下可以筛选到谱带清晰、多态性好的引物22条,可用于枇杷的分子标记分析。     

百合病毒检测芯片的杂交条件优化

分别以感染百合无症病毒、黄瓜花叶病毒和百合斑驳病毒的百合及感染烟草花叶病毒和马铃薯病毒Y的烟草为试材,根据5种植物病毒外壳蛋白基因保守序列设计引物和寡核苷酸探针,并制备基因芯片。用Trizol试剂盒提取感染病毒的植物总RNA,荧光RT-PCR产物与芯片杂交,研究PCR产物是否进行变性处理、杂交时间、杂交温度、杂交液组分SSC和SDS浓度及PCR体系中非荧光引物和荧光引物比例对芯片杂交的影响。结果表明:杂交适宜条件为6×SSC、0.2%SDS的杂交液、42℃杂交60min,PCR体系中非荧光与荧光引物比例为1∶10,PCR产物要进行变性处理。经过整体条件优化后的基因芯片在杂交检测上具有较高的特异性,适于检测百合病毒病。     

岩虱子RAPD反应体系的研究

利用RAPD（随机扩增多态DNA）技术对濒危植物岩虱子进行了RAPD反应体系的研究.以岩虱子叶片为材料,采用改良的CTAB法提取其基因组DNA,进行RAPD反应条件的优化试验.通过单因子试验分别研究了模板DNA用量、Mg^2＋浓度、dNTPS浓度、Taq酶的用量和引物浓度对RAPD反应的影响,建立了适于岩虱子RAPD分析的有效的稳定的反应体系,即在25μL总反应体系中,模板DNA的最适用量为10 ng、Mg^2＋的适宜浓度为2.0 mmol/L、dNTPs的适宜浓度2.2 mmol/L、Taq酶的用量以1 U、随机引物的浓度以2.5 μmol/L为佳.     

Effect of salinity and water table on growth and tolerance of plum and peach

The effects of combinations of salinity (no salt, 2000 p.p.m. or 4000 p.p.m. of CaCl₂ and NaCl, 1:1) and water table (30 cm, 60 cm or 90 cm from the soil surface), on the vegetative growth and tolerance of ‘Golden Japanese’ plum and ‘Mit Ghamre’ and ‘Balady’ peaches were studied. The plants were grown in lysimeters. The growth of the trunk, total shoot length, the increase in shoot length per cm and the fresh weight of top, root and total plant were reduced with increasing salinity of the irrigation water. The effect was accentuated when the plants were maintained at high water table level. The salinity treatments resulted in the death of 43%, 73% and 76% of the plants in the plum, and the ‘Mit Ghamre’ and ‘Balady’ peaches, respectively, indicating that the plum is more tolerant to salinity than the peach. The plants of the salinity treatments showed various symptoms of salt injury, such as leaf burn, defoliation, shoot die-back and finally death. In the peaches, salt injury started to occur in the first growing-season, whereas the salt injury appeared in the plum in the second growing-season. The symptoms were more pronounced in the 4000 p.p.m. treatment than in the 2000 p.p.m. treatment and were more pronounced at the high water table level. The salinity level was the predominant factor and the effect of the water table on the vegetative growth diminished with increase in the salinity level of the irrigation water.     

洞庭1号无籽西瓜套种棉花高产高效模式的研究与应用

根据西瓜、棉花两者生长发育特点进行套种试验,研究其高产高效栽培模式。试验示范结果表明,以西瓜行株距4.5m×0.65m,棉花行株距1.5m×0.45m的种植密度,西瓜与棉花之间互颉作用最小,西瓜和棉花的产量和产值在3个套种模式中位居第1,在5个处理中经济效益最高。     

Primary culture and identification of mouse PASMCs and effects of hypoxia on proliferation and apoptosis of PASMCs

AIM：To establish a fast, accurate and economical technique for culturing mouse pulmonary arteriolar smooth muscle cells (PASMCs), and to explore the effects of hypoxia on the proliferation and apoptosis of the PASMCs. METHODS：In sterile condition, the pulmonary artery was isolated from the male BALB/c mice by digesting with collagenase I, and the cells were cultured in fetal bovine serum-coated flask. Centrifugal procedure was not used during the cell passage. The cell morphology was observed under an inverted phase-contrast microscope. α-Smooth muscle actin was identified by immunocytochemistry and immunofluorescence. The effects of hypoxia on the proliferation and apoptosis of the PASMCs were detected by CCK-8 assay and TUNEL assay. RESULTS：PASMCs were identified by the methods of immunocytochemistry, immunofluorescence staining and observation of morphology. Unlike the rat PASMCs with typical subcultured peak-vally pattern, the mouse PASMCs showed a lot different without a peak-vally pattern. The cells could be subcultured after 5 d to 7 d and there was 3 to 5 generations depending on the activity of the cells. CCK-8 assay demonstrated that the A values of PASMCs exposed to hypoxia increased after 24 h (P＜0.05) as compared with normoxia. TUNEL result showed that the apoptotic index of the PASMCs in hypoxia decreased after 24 h (P<0.05). CONCLUSION：This technique for obtaining cultured mouse PASMCs is simple, fast, accurate and economical. The digestion time is easy to control. Hypoxia promotes the proliferation and inhibits the apoptosis of PASMCs.     

小檗碱和壳聚糖抗蔬菜病原真菌活性测定及复合膜制备

研究了小檗碱和壳聚糖对几种常见蔬菜病原真菌的抑制活性,以及以小檗碱和壳聚糖为主料制备复合膜的方法,并测定了该膜的药物释放效果。试验表明:低浓度(0.234 mg/mL)小檗碱即可抑制辣椒炭疽病菌(Vermicularia capsici)等5种蔬菜病原真菌的生长。20 mg/mL浓度壳聚糖对番茄灰霉病菌(Botrytis cinerea)的抑制率高达65%,而对其余4种果蔬病原真菌也有一定的抑制作用。为了集成这2种天然化合物的优点,制备了小檗碱-壳聚糖复合膜,该膜具有缓释功能,在模拟外部环境(磷酸缓冲液,pH 6.8)条件下,20 d小檗碱累计释放率接近25%,提示其在果蔬贮藏抗菌中的应用价值。     

西瓜与粮油菜套种栽培试验

近年来,我省西瓜发展速度较快,到2004年全省西瓜栽培面积已达3万hm^2.由于我省耕地面积有限,大幅度扩大西瓜单种面积,不仅造成我省西瓜与粮油菜争地的矛盾,而且土地也未能得到充分有效的利用.为了解决这一生产中存在的实际问题,我们从2002年开始,进行了西瓜与粮油菜套种的栽培试验.历经4年的试验、示范,取得了显著的经济效益和社会效益.现将试验结果报告如下.     

钾肥在小白菜和萝卜上的施用效果

采用盆栽试验方法研究了施用钾肥对小白菜、萝卜生长发育、产量、品质及钾素含量的影响。结果表明 ,施用钾肥对小白菜和水萝卜的株高、叶片数无显著的影响 ,但可提高水萝卜块根的产量 ,提高幅度为 31.0 4 %～ 97.93%。施用钾肥可提高小白菜和萝卜的蛋白质和VC含量 ,降低硝酸盐含量 ,增加植株的钾素含量     

世界及中国鳄梨产销概况与展望

依据联合国粮食及农业组织(FAO)数据库、联合国商品贸易统计数据库(UN Comtrade)中的数据,围绕鳄梨种植面积、产量、迚出口市场、主产国生产情况、主要迚出口国贸易情况等斱面,对世界及中国鳄梨的生产与贸易迚行回顾、分析与展望,幵在此基础上,对中国鳄梨产业的发展提出建议。     

Storability of onion seeds and effects of packaging and storage conditions on viability and vigour

Fresh onion seeds desiccated to 6.0% seed moisture content (SMC) were stored in various packaging materials under different storage conditions. Seeds packed in aluminum-laminated pouches beside those stored with silica gel at 25 °C maintained satisfactory germinability and vigour after 12 months. Desiccated seeds stored in moisture impervious containers produced more vigourous seedlings. Germination potential of onion seeds increased with reduced SMC besides storage in moisture impervious packets along with desiccants as physiological and biochemical attributes are regulated. Seed viability and vigour decreased with accelerated ageing due to increased lipid peroxidation, decreased activities of several free radical and peroxide scavenging enzymes. Electrical conductance of seed leachates also increased with ageing. Thus, adoption of appropriate storage temperature and moisture control techniques would significantly affect onion seed quality, which was due to minimum accumulation of free peroxide radicals and enhanced activity of free radical scavenging enzymes.     

遮阴及地膜覆盖对黄皮果实表面温湿度变化及果实品质的影响

以金鸡心黄皮为试材,研究了遮阴及地膜覆盖处理对黄皮果实品质与果实表面温湿度变化的影响。结果表明：阴雨天气下,遮阴及遮阴＋地膜覆盖处理的果实表面日平均湿度低;晴天天气下,地膜覆盖处理及对照果实表面的日平均温度高,昼夜温差大,阴雨及晴天天气下温度变化、湿度变化趋势基本一致;地膜覆盖提高了果实品质,遮阴及遮阴＋地膜覆盖降低了果实好果率。     

热处理对果蔬采后品质及病虫害的影响

采后热处理是近年来快速发展的果蔬采后处理技术,广泛应用于改善果蔬贮藏期间的品质、增强抗逆性等 方面。从果蔬贮藏期间的生理生化、采后品质及防治病虫害等方面阐述了热处理的机理,讨论了热处理对果蔬衰老 方面的生理特征如色泽、风味、硬度(软化)、失重、呼吸、乙烯释放、酶活性等变化及与抗逆性有关的诱导蛋白质合成 及基因表达的影响,较全面阐述了热处理防治果蔬贮藏期间病虫害的机理如直接作用于病菌、害虫与提高果蔬的抗 病虫性等。