套袋与未套袋苹果果实表皮及心部真菌种群结构与变化动态

2011年在山东海阳,2013年在蓬莱和栖霞选取当地常规管理的苹果园,采用传统组织分离法对套袋苹果、未套袋苹果的果实表皮及心部组织进行真菌分离并进行形态学和分子生物学鉴定。对不同时期采集的套袋及未套袋果实表皮与心部的真菌种类、菌落数量、组织分离率、多样性、相似性系数、相对分离频率等指标进行统计分析。结果表明,两个年度内从果实表皮共分离获得真菌43属,从心部获得真菌31属,相对分离频率较高的真菌包括：链格孢Alternaria spp.、枝状枝孢Cladosporium cladosporioides、镰孢菌Fusarium spp.、青霉菌Penicillium spp.等。研究结果表明：果实套袋后,在果实表皮定殖真菌的种类减少,多样性降低,而数量增加,组织带菌率升高;随果实生长,套袋果实表皮上的真菌种类与未套袋果实表皮真菌种类的相似度逐渐降低;7、8月份,套袋果实表皮上真菌种类明显减少,至8月底,套袋果实表面只有链格孢等少数几种优势真菌定殖。果实套袋后,定殖于果实心部的真菌种类和数量有所波动,套袋果和未套袋果上所分离真菌的相似度有所降低;5月底自果实心部分离真菌的种类和数量与9月底所分离真菌的种类和数量差异不大。     

渭北旱塬苹果园地产量和深层土壤水分效应模拟

为了研究实时气象条件下渭北旱塬不同生长年限苹果园地产量变化趋势和深层土壤水分变化规律,在模型适用性与模拟精度验证基础上,应用WinEPIC模型模拟研究了1962—2001年期间洛川旱塬苹果园地产量演变动态和深层土壤水分效应。结果表明：(1) 在模拟研究期间,洛川旱塬4—40年生苹果园产量整体上呈波动性下降趋势,初期产量逐渐增加,11—23年生达到最大值(平均为28.8 t/hm2),之后随降水量年际波动呈现出明显的波动性降低趋势。(2) 40年间苹果园地遭受的干旱胁迫日数呈波动性上升趋势,与年降水量波动趋势相反。(3) 1—15年生期间苹果园地平均年耗水量高于同期年降水量,导致苹果园地0—10 m土层土壤强烈干燥化,逐月土壤有效含水量波动性降低,1—10年生、11—20年生和21—40年生期间发生土壤干燥化并且程度逐渐加剧,但干燥化速率逐渐减缓,土壤干燥化速率分别为95.4 mm/a、12 mm/a和1.5 mm/a。(4) 随生长年限的延长,苹果园地0—10 m土层土壤湿度逐渐降低、土壤干层分布深度逐渐加大,在14年生时超过了10 m,20年生以后2—10 m 土层形成稳定的土壤干层。因此,基于土壤水分利用的苹果生长与果园利用的合理年限为20 a,最长不宜超过23 a。     

苹果柠檬酸合酶3基因家族成员鉴定及表达分析

柠檬酸合酶(citrate synthase 3, CS3)是细胞代谢途径中的关键酶之一,其活性调节着生物体的物质和能量代谢过程。本研究旨在从苹果全基因组中鉴定CS3基因家族成员,并进行生物信息学和表达模式分析,为研究苹果CS3基因的潜在功能提供理论基础。利用BLASTp基于GDR数据库鉴定苹果CS3家族成员,通过Pfam、SMART、MEGA5.0、clustalx.exe、ExPASy Proteomics Server、MEGAX、SOPMA、MEME和WoLF PSORT等软件分析CS3蛋白序列基本信息、亚细胞定位情况、结构域组成、系统进化关系以及染色体定位情况。利用酸含量的测定和实时荧光定量PCR (real-time fluorescence quantitative polymerase chain reaction, qRT-PCR)技术检测苹果6个CS3的组织表达和诱导表达特性。苹果CS3基因家族包含6个成员,这些CS3蛋白包括473−608个不等的氨基酸残基,等电点分布在7.21−8.82。亚细胞定位结果显示CS3蛋白分别定位在线粒体和叶绿体。系统进化分析可将其分为3类,各亚家族基因数量分别为2个。染色体定位结果显示,CS3基因分布在苹果不同的染色体上。蛋白二级结构以a-螺旋为主,其次是无规则卷曲,b-转角所占比例最小。筛选的6个家族成员在不同苹果组织中均有表达,整体表达趋势从高到低依次为MdCS3.4相对表达含量最高,MdCS3.6次之,其他家族成员相对表达量依次为MdCS3.3>MdCS3.2>MdCS3.1>MdCS3.5。qRT-PCR结果显示,MdCS3.1和MdCS3.3基因在酸含量较低的‘成纪1号’果肉中相对表达量最高,酸含量较高的‘艾斯达’果肉中MdCS3.2和MdCS3.3基因相对表达量最高。因此,本研究对不同苹果品种中CS3基因相对表达量进行了检测,并分析了其在苹果果实酸合成过程中的作用。结果表明,CS3基因在不同苹果品种中的相对表达量存在差异,为后续研究苹果品质形成机制提供了参考。     

苹果园中凹唇壁蜂和紫壁蜂的生态位比较研究

本文对凹唇壁蜂Osmia excavata Alfken和紫壁蜂O.jacoti Cockerell的时间、空间、营养和筑巢生态位进行了研究。结果表明,在苹果园内人工释放和管理条件下,两种壁蜂在不同的资源序列上具有不同的生态位宽度、生态位重叠和不同程度的竞争关系。其中在营养上的竞争最为激烈,生态位重叠值和种间竞争系数分别高达0.9690和0.9994。日活动时间生态位竞争次之,其重叠值和种间竞争系数分别为0.7960和0.9350;时间生态位(季节)和空间生态位重叠值则分别为0.6500和0.6710,种间竞争系数为0.8213和0.8234;并且在人工巢管的选择上竞争性最小,生态位重叠值仅为0.4930,种间竞争系数为0.6052。凹唇壁蜂在营养生态位和时间生态位上较高的专一性均表现出其比紫壁蜂对苹果传粉作用更为显著。紫壁蜂在苹果开花后期的传粉作用较强。     

聚丙烯酰胺对坡地苹果园水土流失和土壤养分流失的影响

为了改善坡地苹果园的土壤环境,遏制水土流失和土壤养分流失,探寻聚丙烯酰胺适宜的干撒施用量,2010—2012年在陕北丘陵沟壑区的坡地苹果园,以不施聚丙烯酰胺为对照,分别撒施0.6、0.8、1.0、1.2、1.4和1.6 g·m^-2的聚丙烯酰胺,监测果园地表产流、土壤流失、养分流失和苹果植株生长状况.结果表明: 坡地果园的径流量和5—7月的产流次数均随聚丙烯酰胺撒施量的增加呈“V”型变化,其中撒施量为1.0 g·m^-2时最低,但果园侵蚀泥沙量则随聚丙烯酰胺撒施量的增加而降低.地表径流和侵蚀泥沙中的铵态氮、速效磷和速效钾的浓度均随聚丙烯酰胺撒施量的增加而降低;聚丙烯酰胺可显著降低地表径流中的硝态氮含量,对侵蚀泥沙中的硝态氮含量则无显著影响;侵蚀泥沙中的有机质、全氮、全磷和全钾含量均随聚丙烯酰胺撒施量的增加而降低.聚丙烯酰胺提高了苹果单果质量及产量,但对苹果植株生长及果实风味品质无显著影响.聚丙烯酰胺在坡地苹果园中的适宜撒施量应为1.0 g·m^-2.     

三种拮抗酵母菌对苹果采后青霉病的抑制效果

从苹果果实上分离获得的50余种酵母菌中筛选出了能够有效地抑制苹果青霉病（Peniclium expansum Link)的丝孢酵母（Trichosporon pullulans(Lindner.)Diddens and Lodder)。罗伦隐球酵母（Cryptococcus laurentii(Kuffer.)skin-ner)和粘红酵母（Rhodotorula glutinis(Fresen.)F.C.Harrison)。其中,抑病效果最好的T.pullulans是一种用于采后果实生物防治的新型拮抗菌,研究了这三种拮抗菌不同浓度处理和外加营养物质以及与钙配合使用对苹果青霉病的抑病效果。实验结果表明;酵母菌浓度越高,抑病作用越强;外源营养物质的加入削弱了酵母菌的拮抗效果;在C.laurentii的细胞悬浮液中加入0.18mol/L的CaCl2能显著提高其抑病能力。但增加CaCl2对T.pullulans和R.glutinis的抑病效果却没有明显作用。     

Post-translational inhibitory regulation of acid invertase induced by fructose and glucose in developing apple fruit

Acid invertase (EC 3.2.1.26) is one of the key enzymes involved in the carbohydrate sink-organ development and the sink strength modulation in crops. The experiment conducted with 'Starkrimson' apple (Malus domestica Borkh) fruit showed that, during the fruit development, the activity of acid invertase gradually declined concomitantly with the progressive accumulation of fructose, glucose and sucrose, while Western blotting assay of acid invertase detected a 30 ku peptide of which the immuno-signal intensity increased during the fruit development. The im-muno-localization via immunogold electron microscopy showed that, on the one hand, acid invertase was mainly located on the flesh cell wall with numbers of the immunosignals present in the vacuole at the late stage of fruit development; and on the other hand, the amount of acid invertase increased during fruit development, which was consistent with the results of Western blotting. The in vivo pre-incubation of fruit discs with soluble sugars showed that     

Epidemiology of apple proliferation (AP) in northwestern Italy: evaluation of the frequency of AP-positive psyllids in naturally infected populations of Cacopsylla melanoneura (Homoptera: Psyllidae)

Adults of Cacopsylla melanoneura, vector of the apple proliferation (AP) phytoplasma, were collected every 2 weeks from January until May in 2000 and 2001 by the beating tray method in eight apple orchards of the Aosta Valley (northwestern Italy). Total DNA was extracted from batches of five insects and amplified with the universal phytoplasma primers P1/P7 in direct PCR. A nested PCR assay was then performed on P1/P7 amplicons using the primers fO1/rO1, specific for the AP‐ phytoplasma group. The digestion of fO1/rO1 amplicons with Ssp I restriction endonuclease confirmed that C. melanoneura adults harboured the AP phytoplasma. The data obtained with PCR were used to estimate the proportion of AP‐positive insects in over wintered and offspring adults. Percentages of AP‐positive insects of 3.6% and 0.8% were estimated in 2000 among over wintered and offspring psyllids respectively. In 2001 only the over wintered insects were found infected, with an estimated proportion of 2.8%. The seasonal abundance of the vector was measured using yellow sticky traps. C. melanoneura was always present at a low population level, and the highest density was recorded from mid‐February until mid‐March in both years. The results show that the overwintered population is higher and spends a longer period in apple orchards, suggesting the crucial role of the overwintered adults in vectoring AP.     

酿酒葡萄皮渣中白藜芦醇的抗氧化活性研究

本文利用甲醇溶剂对酿酒葡萄皮渣进行浸提,得到白藜芦醇粗提物.粗提物通过硅胶柱层析进行纯化.然后,研究了白藜芦醇的氧自由基、超氧阴离子自由基、羟自由基、DPPH自由基的清除能力.结果显示:白藜芦醇具有很强的自由基清除能力和抗氧化能力,并且随着浓度的增加,抗氧化能力增强.     

Isolation and pathogenicity of some South African entomopathogenic fungi (Ascomycota) against eggs and larvae of Cydia pomonella (Lepidoptera: Tortricidae)

Virulent entomopathogenic fungal strains against Cydia pomonella (Lepidoptera: Tortricidae) were isolated and identified in the Western Cape Province of South Africa. Thirty-nine isolates belonging to six species were obtained using the insect bait method. Generally, Metarhizium robertsii (Ascomycota: Hypocreales) was the most frequently encountered species representing 51% of the total number of isolates collected from the soil samples. This is the first report of M. robertsii from southern Africa. Mortality data from an immersion bioassay indicated that the 39 fungal isolates were pathogenic against fifth instar larvae of C. pomonella inducing 47–85% insect mortalities. Two M. robertsii strains, MTL151 and GW461, induced 85% larval mortality and were selected for further evaluations. The exposure of freshly laid eggs to wax papers that were pre-treated with fungal spores ranging from 10³ to 10⁸ spores ml^?1 of MTL151 and GW461 resulted in a significant reduction of egg hatchabilities, from 95 to 66% and 93 to 71%, respectively as spore concentration increased. First instar larval neonates were exposed to apples that were pre-sprayed topically with varied conidia suspensions (10³?10⁸ spores ml^?1). The mean percentage of participating apples with larvae in their cores/flesh significantly reduced from 53 to 10% (MTL151) and 76 to 10% (GW461) of 10 apples, and a concurrent decrease in the incidence of apple fruit rot was observed as conidia concentration increased. Up to 90% of apples treated with 1 × 10⁸ spores ml^?1 had no larvae present in their cores and this result compared favourably with the commercial pesticide Fruitfly®, containing the active ingredient cypermethrin (20 g/l) used at a recommended dose of 0.25 ml/250 ml of water.