PVY侵染后烟草营养成分的变化及其对介体烟蚜生长发育的影响

【目的】明确马铃薯Y病毒(potato virus Y,PVY)侵染后诱导的烟草营养成分的变化及其对烟蚜Myzus persicae生命特性的影响,旨在进一步解析PVY-烟草-烟蚜三者间的互作机制。【方法】通过蒽酮比色法和氨基酸自动分析仪测定了PVY不同侵染时期烟株体内的可溶性糖和游离氨基酸含量的变化;测定和比较了感病与健康烟草植株上烟蚜种群生长发育、成虫寿命、繁殖力和有翅蚜产生量的差异性。【结果】PVY侵染前、中、后期(分别为侵染后5,12和20 d)的烟草叶片中游离氨基酸的总量均显著高于健康烟草叶片。相较于健康烟草叶片,在PVY侵染前期的烟草叶片中,谷氨酸、脯氨酸、天冬氨酸、色氨酸、缬氨酸、赖氨酸和组氨酸的含量显著增加;PVY侵染中期,感病叶片中丝氨酸含量显著下降,谷氨酸、天冬氨酸、色氨酸、缬氨酸、亮氨酸、苯丙氨酸、精氨酸和组氨酸含量显著提高;PVY侵染后期,感病叶片中甘氨酸含量显著下降,谷氨酸、脯氨酸、天冬氨酸、苏氨酸、缬氨酸、亮氨酸、丙氨酸、苯丙氨酸、组氨酸、酪氨酸和精氨酸含量显著提高。在PVY侵染的前期和中期,感病叶片中的可溶性糖含量显著高于健康烟叶,而在侵染后期感病叶片中可溶性糖含量显著低于健康烟草叶片的。PVY侵染前期和中期的烟草叶片中总糖和总游离氨基酸的含量比值显著高于健康烟草叶片中的。在PVY侵染的烟草植株和健康烟草植株上取食的烟蚜其发育历期、若蚜历期、成蚜繁殖期、繁殖后期、寿命、烟蚜种群的内禀增长率、周限增长率和平均世代周期均无显著差异,但在感病烟草植株上取食的烟蚜成蚜繁殖前期显著缩短,其繁殖力和净生殖率显著提高。相较于健康烟草植株,在PVY侵染烟草植株上定殖的烟蚜种群有翅蚜发生的高峰期提前。【结论】PVY侵染前期和中期提高了寄主烟草的营养品质,从而提高了烟蚜的繁殖力。侵染后期烟草营养品质的下降,促使烟蚜种群有翅蚜的产生和扩散,从而有利于PVY自身的传播。     

硒肥对马铃薯硒素吸收、转化及产量、品质的影响

通过设对照(CK)、保水缓释硒肥(W)、生物炭基硒肥(C)、硒酸钠硒肥(S)4个处理来研究不同硒肥对马铃薯(品种为早大白)硒素吸收、转化及产量、品质的影响。结果表明:各处理马铃薯各器官硒含量在生育期内总体上呈下降趋势,马铃薯各器官的硒含量呈现:苗期根茎叶片;成熟期叶片茎块茎的特点;随着硒肥用量的增加,W处理下的总硒、无机硒、有机硒含量呈增大趋势,产量、有机硒转化率、粗蛋白、还原糖和Vc呈先升高后降低的趋势;C处理和S处理下,马铃薯以上各指标均呈先升高后降低的趋势,在低施硒量(0.126 kg/hm2)时,3种硒肥显著降低了马铃薯块茎淀粉含量,之后随着施硒量的增加淀粉含量变化不显著;与对照相比,3种硒肥在适宜施硒量(0.379 kg/hm2)时,马铃薯产量提高了4.87%—5.44%,粗蛋白含量增加了12.18%—20.03%,还原糖提高了6.45%—12.90%,Vc含量提高-0.54%—3.11%,有机硒转化率增加13.00%—15.10%,淀粉含量增加了-0.73%—1.12%;综合考虑3种硒肥对马铃薯含硒量、产量、品质的影响,W处理最佳,C处理次之,S处理最差。     

低温条件下转AcInv反义基因马铃薯品系的干物质、淀粉和还原糖含量变化

检测低温（4℃）贮藏条件下转AcInv反义基因与否的马铃薯品系块茎中干物质、淀粉和还原糖含量变化的结果表明：随着低温贮藏时间的延长,各转基因品系块茎中干物质含量变化不大,淀粉含量虽有下降但较为平缓;转基因品系中的还原糖含量增加幅度比受体品种的明显降低,转Aclnv反义基因的‘甘衣薯1号’和‘大西洋’块茎抗低温糖化能力明显高于未转AcInv反义基因的品系。     

反义酸性转化酶基因低温贮藏马铃薯块茎还原糖和淀粉含量的影响

对2个含有酸性转化酶(AcInv)反义基因的转基因马铃薯品系及对照品种进行低温贮藏(4℃)及室温还暖处理.随低温贮藏时间的延长,供试材料均表现出还原糖含量升高,总淀粉含量下降的趋势.低温处理40 d时,"Ac转Atlantic"和"Ac转甘农薯2号"的还原糖含量比未转基因品种低23%和18%.总淀粉含量分别比未处理前下降约1.0%和1.3%,支链淀粉含量分别下降约1.4%和1.7%,淀粉直/支比明显低于对照,分别为0.29和0.38.块茎的石蜡切片显示,转基因块茎中深蓝色淀粉颗粒明显少于未转基因对照.另外,对低温贮藏的块茎室温还暖后,2个转基因品系的还原糖含量仍低于对照品种.实验结果证明反义AcInv基因对低温贮藏下块茎还原糖和淀粉含量具有下向调节作用.     

番茄环纹斑点病毒与马铃薯Y病毒复合侵染烟草的细胞病理特征

在自然情况下, 番茄环纹斑点病毒(TZSV)和马铃薯Y病毒(PVY)通常复合侵染同一植株。该文首次报道了TZSV和PVY复合侵染烟草(Nicotiana sp.)植株的细胞病理特征, 并与单独侵染进行了比较分析。复合侵染的烟草植株细胞中, TZSV病毒颗粒明显增多, 并聚集于囊膜内形成包涵体, 与PVY的风轮状及片层状内含体和病毒颗粒聚集体交叉分布于细胞质(内含线粒体明显增多)中, 线粒体、叶绿体和细胞核结构较完整; 两种病毒的颗粒、包涵体和内含体数量均较单一侵染增多, 且对寄主亚细胞结构的破坏较单一侵染为轻, TZSV和PVY及其与寄主的互作可能存在协生作用。     

一种适合于马铃薯病毒ssRNA快速提取技术

针对马铃薯病毒ssRNA制样过程中产生色素、多酚物质抑制RT PCR反应的问题 ,通过确定多酚氧化酶抑制剂Na2 SO3 的最适浓度 ,筛选核酸快速浸提的表面活性剂 ,研制出马铃薯病毒简易浸提制样技术 ,可以从植物组织中快速释放ssRNA并减少PCR反应抑制物质的析出。此法简便有效 ,可以从马铃薯叶片 ,叶柄 ,茎和块茎中快速提取PVY、PVX ,PLRV、PVA、PVS等多种马铃薯病毒ssRNA ,适合于同步验证马铃薯原原种和原种的脱毒效果以及田间疑似病症的快速诊断和不同马铃薯病毒的准确鉴定。     

表达PVY和PLRV双价外壳蛋白基因马铃薯的抗病性研究

表达马铃薯Y病毒(PVY)和马铃薯卷叶病毒(PLRV)双价外壳蛋白基因的马铃薯(Solanum tubero-sum L.)栽培品种“Favorita”和“虎头”,经摩擦接种PVY和用桃蚜接种PLRV后,观察症状并用ELISA测定病毒滴度。结果表明,两个品种转双价CP基因的各株系,接种病毒后表现无症状或症状轻微,其中PVY和PLRV平均滴度均较不转基因对照植株低。不同品种对PVY和PLRV的抗性比较表明,转双价CP基因的“Favorita”对PVY抗性较明显,而转双价CP基因的“虎头”则对PLRV抗性较对PVY抗性明显。不同转基因株系抗病毒水平不同。“Favorita”9个转双价CP基因株系中有6个株系PVY滴度较未转基因对照降低52.5％～90.0％,而“虎头”7个转双价CP基因株系中有4个株系PLRV含量较对照降低53.0％～98.0％。在抗性株系中还出现一些抗1种病毒或抗2种病毒的抗性较强的单株。     

马铃薯AGPase活力反馈调控光合速率定量分析

为了揭示ADP-葡萄糖焦磷酸化酶(AGPase)改变对叶片光合作用的反馈调控,以改变AGPase活力的马铃薯转化株系为材料,测定了其AGPase活力(AA)、块茎淀粉积累量(SC)、叶片光合速率(PR)、叶绿素(CC)和蔗糖含量(SUC)等;测定结果分析显示,各株系的AGPase活力(AA)、块茎淀粉积累量(SC)和叶片光合速率(PR)间存在显著差异,且均显著高于对照;株系间AGPase活力、块茎淀粉含量和叶片光合速率呈显著正相关性;每单位AGPase活力上升可贡献贮藏器官淀粉积累增加2.5%,且是影响叶片光合速率的重要因素;结果表明,AGPase不仅是下游淀粉积累的限速酶,而且可向上游反馈调控光合速率。     

纳米硒对紫色马铃薯生长及其矿质元素含量和品质特性的影响

以紫色马铃薯品种‘黑美人’为试材,采用水培法,分别用不同浓度(0、0.38、0.19和0.095 mmol/L)纳米硒溶液对紫色马铃薯进行叶面喷施处理,研究纳米硒对紫色马铃薯生长、矿质元素含量及品质特性的影响。结果表明:(1)与对照相比,各硒处理马铃薯的生物量与单株结薯数均显著增加,其中喷施0.095 mmol/L硒处理的生物量最高且增幅达1.5倍,喷施0.19 mmol/L硒处理的单株结薯数显著增加2.2倍。(2)纳米硒能够显著提高紫色马铃薯叶片、根系、块茎硒含量,各器官硒含量大小呈现:根系>叶片>块茎的特点,且喷施0.095 mmol/L硒处理块茎总硒含量达0.106 mg/kg,较对照显著提高0.65倍,达到了马铃薯块茎的富硒标准;同时纳米硒可在不同程度上调控K、Ca、Mg、Mn、Zn在马铃薯各器官中的分配。(3)随施硒浓度的增加,紫色马铃薯块茎中淀粉、可溶性蛋白及游离氨基酸含量呈先增加后降低的变化趋势,均在喷施0.19 mmol/L硒处理下达到最大值,且较对照分别显著增加56.33%、26.91%和27.89%;块茎中花青素、可溶性糖含量呈下降趋势,均在喷施0.095 mmol/L硒处理下达到最大值,且较对照分别显著提高24.73%、25.33%;而块茎中硝态氮含量呈上升趋势,在喷施0.095 mmol/L硒处理下最低并较对照显著降低34.82%。研究表明,叶面喷施0.095~0.19 mmol/L纳米硒溶液能够显著促进紫色马铃薯生长和单株结薯数,提高硒元素含量,调控矿质元素含量在器官中分配,有效改善其块茎品质特性。     

连作马铃薯植株库源关系及其对块茎产量的调节机理

甘肃中部沿黄灌区是国内重要的加工型马铃薯生产基地和种薯繁殖基地,但集约化生产带来的连作障碍已严重影响到产业的健康发展.为揭示马铃薯连作障碍机理,本研究通过田间试验,设置马铃薯连作年限为0～5年的处理,研究了马铃薯植株库容量、库活性和源活性对连作的响应特征及其对块茎产量的调节机制.结果表明: 短期连作(1～2年)条件下块茎产量较非连作(0年)无显著变化,长期连作(3～5年)显著下降28.6%～32.8%,单薯质量降低是导致块茎产量下降的直接原因.长期连作马铃薯库容量较非连作显著降低38.4%～53.0%.长期连作导致块茎形成推迟,同时单薯干物质积累量降低.长期连作也显著降低马铃薯源活性,与非连作相比,株高、主茎分枝数、叶绿素含量和叶片干物质量均显著下降,根系形态发育受到抑制,根系活力显著下降28.6%～63.1%,叶片RuBP羧化酶和蔗糖磷酸合成酶活性分别显著下降52.6%～64.6%和26.3%～53.4%.长期连作条件下马铃薯源端生产性能降低导致同化产物减少,花后阶段向块茎的输入不足,降低块茎产量.库源关系失衡是甘肃中部沿黄灌区长期连作马铃薯产量大幅降低的原因.     

壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

国内外蝗害治理技术现状与展望

本文首先概述了国内外蝗虫发生与为害的态势,总结了现阶段我国蝗虫发生与为害的主要特点:即农田飞蝗暴发频繁而且严重,草原土蝗的发生时常造成严重的经济损失,而且侵入城市干扰市民生活,我国与周边国家之间蝗虫过境迁移频繁,使用化学农药污染环境和农产品;分析了国内外蝗虫防治对策与技术的发展现状,重点介绍了应急防治和可持续治理对策、...     

Synthesis and turnover of cerebrosides and phosphatidylserine of myelin and microsomal fractions of adult and developing rat brain.

The synthesis and turnover of cerebrosides and phospholipids was followed in microsomal and myelin fractions of developing and adult rat brains after an intracerebral injection of [U-14C]serine. The kinetics of incorporation of radioactivity into microsomal and myelin cerebrosides indicate the possibility of a precursor-product relationship between cerebrosides of these membranes. The specific radioactivity of myelin cerebrosides was corrected for the deposition of newly formed cerebrosides in myelin. Multiphasic curves were obtained for the decline in specific radioactivity of myelin and microsomal cerebrosides, suggesting different cerebroside pools in these membranes. The half-life of the fast turning-over pool of cerebrosides of myelin was 7 and 22 days for the developing and adult rat brain respectively. The half-life of the slowly turning-over pool of myelin cerebrosides was about 145 days for both groups of animals. The half-life of the rapidly turning-over microsomal cerebrosides was calculated to be 20 and 40 h for the developing and adult animals respectively. The half-life of the intermediate and slowly turning-over microsomal cerebrosides was 11 and 60 days respectively, for both groups of animals. The amount of incorporation of radioactivity into microsomal cerebrosides from L-serine was greatly decreased in the adult animals, and greater amounts of the precursor were directed towards the synthesis of phosphatidylserine. In the developing animals, considerable amounts of cerebrosides were synthesized from L-serine, besides phosphatidylserine. The time-course of incorporation indicated that a precursor-product relationship exists between microsomal and myelin phosphatidylserine. The half-life of microsomal phosphatidylserine was calculated to be about 8 h for the fast turning-over pool in both groups of animals.     

Identification and composition of the tonsillar and anal enterococcal and streptococcal flora of dogs and cats

Enterococcus faecalis was the most frequently isolated enterococcal species from anal swabs and tonsils of dogs and cats, although in the anal samples from dogs Ent. hirae was found almost as often as Ent. faecalis. Most Ent.faecium strains from dog tonsils differed from those associated with humans and other animals in that they fermented sorbitol. Typical Ent. avium as well as atypical Ent. avium -like strains were seen in dogs, while the related species Ent. raffinosus was associated with cat tonsils. Enterococcus cecorum also occurred mainly in cats. Certain atypical strains, presumptively identified as Ent. cecorum , shared characteristics with Ent. columbae.
The most frequent streptococcal species in tonsils of cats and dogs were Streptococcus suis and Strep. canis. Streptococcus canis and Strep. bovis predominated in anal swabs. The canine Strep. suis differed from the common porcine strains in fermenting mannitol.
Forty-seven of the 288 isolates examined could not be identified or related to known species. The characteristics of two groups of these bacteria, provisionally called 'Ton 31 group' and 'O7 group' are described.     

Molecular characterisation of species and genotypes of Cryptosporidium and Giardia and assessment of zoonotic transmission

The molecular characterisation of species and genotypes of Cryptosporidium and Giardia is essential for accurately identifying organisms and assessing zoonotic transmission. Results of recent molecular epidemiological studies strongly suggest that zoonotic transmission plays an important role in cryptosporidiosis epidemiology. In such cases the most prevalent zoonotic species is Cryptosporidium parvum. Genotyping and subtyping data suggest that zoonotic transmission is not as prevalent in the epidemiology of giardiasis. Molecular characterisation of Cryptosporidium and Giardia is a relatively recent application that is evolving as new genes are found that increase the accuracy of identification while discovering a greater diversity of species and yet unnamed taxa within these two important genera. As molecular data accumulate, our understanding of the role of zoonotic transmission in epidemiology and clinical manifestations is becoming clearer.     

白术同源四倍体的诱导和鉴定及其与二倍体过氧化物酶的比较

以白术（Atractylodes macrooephala Koidz.）二倍体组培苗为材料,对其四倍体诱导方法进行研究,共获得45个白术同源四倍体株系,为优良株系的选育提供了材料。此外,还分析比较了其中8个白术四倍体株系与二倍体的过氧化物酶同工酶（POD）的酶谱差异,发现四倍体各株系过氧化物酶同工酶谱比二倍体的均多了Rf0.310的谱带,且总过氧化物酶比活力也发生了很大改变,对探讨白术四倍体优良株系的生理生化机理具有一定的参考价值。     

Kinetics and thermodynamics of peroxidase- and laccase-catalyzed oxidation of N-substituted phenothiazines and phenoxazines

N -substituted phenothiazines (PTs) and phenoxazines (POs) catalyzed by fungal Coprinus cinereus peroxidase and Polyporus pinsitus laccase were investigated at pH 4–10. In the case of peroxidase, an apparent bimolecular rate constant (expressed as k _cat/K _m) varied from 1 ×10⁷M⁻¹s⁻¹to 2.6×10⁸M⁻¹s⁻¹ at pH 7.0. The constants for PO oxidation were higher in comparison to PT. pH dependence revealed two or three ionizable groups with pK _a values of 4.9–5.7 and 7.7–9.7 that significantly affected the activity of peroxidase. Single-turnover experiments showed that the limiting step of PT oxidation was reduction of compound II and second-order rate constants were obtained which were consistent with the constants at steady-state conditions. Laccase-catalyzed PT and PO oxidation rates were lower; apparent bimolecular rate constants varied from 1.8×10⁵M⁻¹s⁻¹ to 2.0×10⁷M⁻¹s⁻¹ at pH 5.3. PO constants were higher in comparison to PT, as was the case with peroxidase. The dependence of the apparent bimolecular constants of compound II or copper type 1 reduction, in the case of peroxidase or laccase, respectively, was analyzed in the framework of the Marcus outer-sphere electron-transfer theory. Peroxidase-catalyzed reactions with PT, as well as PO, fitted the same hyperbolic dependence with a maximal oxidation rate of 1.6×10⁸M⁻¹s⁻¹ and a reorganization energy of 0.30 eV. The respective parameters for laccase were 5.0×10⁷M⁻¹s⁻¹ and 0.29 eV. Received: 20 September 1999 / Accepted: 24 February 2000