Hydrolytic Enzyme Inhibitors and Necrotic Reactions in Potato Leaves Reduce Reproductive Success of Colorado Potato Beetle

In field and laboratory conditions the reproductive potential （survival at different stages of development） of the Colorado potato beetle has been measured, the dependence of the parameters of insects fertility on varietal features of the feed has been investigated, the food activity of the representatives of insects from four areas of the Southern Urals has been measured, the intensity and time of hypersensitivity reactions on potato leaves has been defined. In laboratory experiments, the level of activity of hydrolytic enzymes in insects and the level of activity of these enzymes inhibitors in the leaves of three potato varieties differing in their resistance to the Colorado potato beetle have been determined. The results show that the insects-representatives of different samples have different food activity in relation to different potato varieties. High mortality rates were revealed at feeding with leaves of potato of the Bashkir variety characterized by a high level of hydrolases inhibitors and relatively rapid development of necrotic reactions after the appearance of eggs laying on the leaves. We suppose that the neerogenetic barrier appearing in the form of necrosis is the major cause of the fetal mortality of insects. The level of the content of inhibitors inactivating larvae digestive enzymes affects the survival of insects at postembryonic stages. The speed of the necrosis emergence, the level of activity of digestive enzymes inhibitors in potato can be considered as criteria for the selection of breeding material for resistance to the Colorado potato beetle.     

胰蛋白酶基因的系统发育分析

下载NCBI数据库中的动物胰蛋白酶基因的氨基酸序列,用CLUSTAL X2程序进行序列比对,用MEGA3.1软件中的Neighbor-joining法构建系统发育树.序列分析表明,胰蛋白酶氨基酸有多处保守基序,且保守性较高,最典型的保守序列有催化三联体结构:His(组氨酸)-Asp(天冬氨酸)-Ser(丝氨酸)以及金有6个保守的半胱氨酸形成3对肽内二硫键.另外,此序列还具有特异性的底物结合位点,这就决定了胰蛋白酶的专一性.系统发育分析结果显示,脊椎动物与无脊椎动物被分成2个大的分支,而脊椎动物中trypsinX3物种被单独分为一支,这表明typsinX3物种与脊椎动物的同源性较低.     

酶法提取猕猴桃皮和渣中果胶的工艺研究

[目的]去除猕猴桃皮和渣中的淀粉和蛋白质,提取并制备膳食食用果胶。[方法]通过单因素和正交试验设计,分别确定0.4%淀粉酶和0.4%胰蛋白酶提取猕猴桃皮和渣中果胶的适宜工艺条件。[结果]0.4%淀粉酶可有效去除猕猴桃皮和渣中的淀粉,其适宜工艺条件为：料液比1∶10.0,50℃下酶解60 min;0.4%胰蛋白酶可有效去除猕猴桃皮和渣中的蛋白质,其适宜工艺条件为：料液比1∶10.0,35℃下酶解60 min。在最佳工艺条件下,经离心、浓缩得到果胶,其中猕猴桃皮和渣中的果胶得率分别为3.10%和1.39%。[结论]通过改良酶法,建立了猕猴桃皮和渣中果胶提取的适宜工艺。     

不同热处理对黑豆干物质消化率,蛋白质消化率的影响

为了研究热处理的温度和时间对黑豆干物质消化率(DMD)、蛋白质消化率(PD)及脲酶活性的影响规律,对经过110℃和121℃下分别处理3,9和15min的饲用黑豆进行了离体干物质消化率和离体蛋白质消化率及脲酶活性的测定。结果表明:在121℃处理15min的样本干物质消化率和蛋白质消化率最高,分别为75.13%,90.91%,脲酶活性最低,为0.04mg/min·g。作者认为:在本研究范围内,湿热处理最适宜的温度是121℃,时间是15min。     

Ontogenetic development of digestive enzymes and effect of starvation in miiuy croaker <Emphasis Type="Italic">Miichthys miiuy</Emphasis> larvae

The ontogenetic development of the digestive enzymes amylase, lipase, trypsin, and alkaline phosphatase and the effect of starvation in miiuy croaker Miichthys miiuy larvae were studied. The activities of these enzymes were detected prior to exogenous feeding, but their developmental patterns differed remarkably. Trypsin activity continuously increased from 2 days after hatching (dah), peaked on 20 dah, and decreased to 25 dah at weaning. Alkaline phosphatase activity oscillated at low levels within a small range after the first feeding on 3 dah. In contrast, amylase and lipase activities followed the general developmental pattern that has been characterized in fish larvae, with a succession of increases or decreases. Amylase, lipase, and trypsin activities generally started to increase or decrease at transitions from endogenous to exogenous feeding or diet changes, suggesting that these enzymatic activities can be modulated by feeding modes. The activities of all the enzymes remained stable from 25 dah onwards, coinciding with the formation of gastric glands and pyloric caecum. These results imply that specific activities of these enzymes underwent changes due to morphological and physiological modifications or diet shift during larval development but that they became stable after the development of the digestive organs and associated glands was fully completed and the organs/glands functioned. Trypsin and alkaline phosphatase were more sensitive to starvation than amylase and lipase because delayed feeding up to 2 days after mouth opening was able to adversely affect their activities. Enzyme activities did not significantly differ among feeding groups during endogenous feeding; however, all activities were remarkably reduced when delayed feeding was within 3 days after mouth opening. Initiation of larvae feeding should occur within 2 days after mouth opening so that good growth and survival can be obtained in the culture.     

水稻抽穗后喷施多胺合成抑制剂对水稻籽粒多胺含量与组成的影响

以高多胺水稻品种金陵香糯为材料,在抽穗期、开花期和灌浆期用不同浓度的多胺合成抑制剂D 精氨酸（D Arg）、甲基乙二醛双（鸟苷腙）（MGBG）和环己胺（CHA）喷施穗部和剑叶,研究它们对水稻籽粒中游离态和结合态多胺含量和组成的影响。结果表明,D Arg对3个时期水稻籽粒中游离态和结合态各多胺含量及总量均有显著抑制作用,其中开花期和灌浆期处理的效果较明显。MGBG和CHA主要影响籽粒多胺的组成,对多胺总量的影响仅MGBG在开花期、CHA在抽穗期和开花期处理的抑制作用达显著水平。     

改性尿素硝酸铵溶液调控氮素挥发和淋溶的研究

为了提高肥料的利用率,以尿素硝酸铵溶液为原料、聚氨酸为保护剂,复合抑制剂NBPT(N-丁基硫代磷酰三胺)和DMPP(3,4-二甲基吡唑磷酸盐)为材料,开发出改性尿素硝酸铵溶液(YUL1和YUL2),研究其对华北平原夏玉米追肥过程中的氨挥发和淋溶损失的调控效果。田间试验设置6个处理:不施氮肥(CK)、农民习惯追施尿素(CN)、优化追施尿素(CNU)、优化追施尿素硝酸铵溶液(UAN)、优化追施改性尿素硝酸铵溶液(YUL1)和优化追施改性尿素硝酸铵溶液(YUL2)。采用扫描电镜和能谱仪分析相关指标变化,在夏玉米喇叭口期追施氮肥后15d内进行田间原位连续动态观测氨挥发和土壤铵态氮和硝态氮变化,并在玉米成熟期测定产量,计算经济效益。结果表明,改性尿素硝酸铵溶液清澈无杂质,流延后成膜表面光滑、致密,抑制剂在膜表面分布均匀;能谱测试膜层表面磷硫含量增高,证明复合抑制剂与尿素硝酸铵溶液达到有效融合。在同等优化施氮量下:与CNU相比, YUL1氨挥发总量显著降低19.3%, YUL2增加9.6%;与UAN相比, YUL1、YUL2分别显著降低57.3%和42.0%。与其他施氮处理相比, YUL1和YUL2夏玉米季生长中后期0～20 cm土层依然保持相对较高的氮素含量水平,夏玉米收获后土壤硝态氮含量分别比CNU高46.0%和43.4%,比UAN高45.6%和44.7%;180～200cm土层硝态氮含量显著低于其他处理。在保证产量和净收益的同时,改性尿素硝酸铵肥料显著降低了氮素的氨挥发和淋溶损失浓度,尿酶抑制剂含量相对较高的YUL1抑制氨挥发的效果更好,硝化抑制剂含量相对高的YUL2硝态氮向下淋失的风险更小。     

昆虫取食和剪叶刺激对油松针叶内部分防御物质的诱导效应

为了研究自然条件下昆虫取食及剪叶刺激对油松诱导抗性的影响,本试验应用香草醛—盐酸法,亚硝酸钠—硝酸铝比色法以及紫外分光光度法,分析了剪叶和不同程度油松毛虫取食处理后,混交林和纯林中油松针叶内部分次生代谢物和蛋白酶抑制剂活性的动态变化。结果表明:剪叶及不同程度油松毛虫取食能够诱导缩合单宁、黄酮含量和胰蛋白酶抑制剂、胰凝乳蛋白酶抑制剂活性增加。与对照相比,胰凝乳蛋白酶抑制剂在混交林中的活性比纯林更为明显。说明剪叶及昆虫取食可诱导增加油松针叶内的缩合单宁和黄酮含量、提高胰蛋白酶抑制剂和胰凝乳蛋白酶抑制剂的活性,进而增强油松的抗虫性,林分类型可在一定程度上影响油松诱导防御物质的变化。     

Aggressiveness of Xanthomonas axonopodis pv. glycines isolates to soybean and hypersensitivity responses by other plants

Xanthomonas axonopodis pv. glycines ( Xag ) causes bacterial pustule disease which can significantly reduce the production of soybean. A collection of 26 isolates of Xag from different soybean-production areas of Thailand was shown to differ with regard to aggressiveness on soybean. They also differed in their ability to induce a hypersensitive response (HR) on four cultivars of tobacco and on other plant species including pepper, tomato, cucumber, pea and sesame. Tomato was most sensitive to HR induction by Xag . Isolate KU-P-34017 caused an HR on all the plant species tested. The minimal concentration of KU-P-34017 needed to induce HR on tobacco was approximately 5 × 10⁸ CFU mL⁻¹. A bacterium–plant interaction period of at least 2·5 h was necessary for HR, and different temperatures, relative humidity and light periods did not affect HR development. Inhibitors of eukaryotic metabolism, including cobalt chloride, lanthanum chloride and sodium orthovanadate (completely), and cycloheximide (partially) blocked the HR on tobacco, indicating the association of an active plant response. In contrast, the HR on tomato was inhibited only by cobalt chloride.     

Genetic Control of Resistance to the Piperidine Fungicide Piperalin in Ustilago maydis

Mutants of Ustilago maydis resistant to the piperidine fungicide piperalin were isolated in a mutation frequency of 2.4 × 10^–5 after UV-irradiation and selection on media containing 50gml^–1 piperalin. Genetic analysis with 15 such mutant isolates resulted in the identifications of two unlinked chromosomal loci, the U/ppl-1 locus with two allelic genes (U/ppl-1A and U/ppl-1B) and the U/ppl-2 locus. The U/ppl-2 and U/ppl-1A mutations are responsible for two levels of moderate and high resistance to piperalin (resistance factor, Rf: 54 and 135, respectively, based on effective concentration causing a 50% reduction in the growth rate, EC₅₀), while the U/ppl-1B mutation gives only a small reduction (approximately 8-fold) in piperalin sensitivity. Cross-resistance studies with other SBIs shows that the major gene (U/ppl-2 and U/ppl-1A) mutants are resistant to fenpropidin (Rf: 43 and 68), fenpropimorph (Rf: 261 and 283) and tridemorph (Rf: 9 and 10), but not to the inhibitors of C-14 demethylase (DMIs) and squalene epoxidase. The minor gene mutation U/ppl-1B codes a low-level of resistance (approximately 5—12-fold) to the above morpholine-type fungicides, but in contrast with the major gene mutations it increases 2–10 times the sensitivity to triazoles: triadimefon, triadimenol, propiconazole and flusilazole. Crosses between mutants carrying the U/ppl-genes with compatible isolates carrying the U/fpd, U/fpm or U/tdm mutations, which have been identified in previous genetic works for resistance to morpholine-type fungicides, yielded, with the exception of U/ppl-2 ×U/fpm-2 cross, a large number of recombinants with wild-type sensitivity, indicating that the mutant genes involved in these crosses, were not allelic. An additive gene effect was observed only between nonallelic minor genes U/ppl-1B and U/fpm-1B or U/tdm-1,2. Studies of the fitness of piperalin-resistant isolates showed that the reduced sensitivity of major gene mutants was not associated with changes on the phytopathogenic fitness determining characteristics, such as growth in liquid culture and pathogenicity on young corn plants. Conversely, the minor gene mutation U/ppl-1B appeared to be pleiotropic, having significantly adverse effects on the phytopathogenic fitness.