The effect of aging of milk samples on the accuracy of infrared analysis of milk composition]

The effect of a decrease (and/or fermentation) in the lactose content during milk storage under different conditions was investigated on the accuracy of the results obtained on a Milko-Scan apparatus to contribute to the present knowledge of this problem. The results were in agreement with some results cited in the literature. These wavelengths are used for infrared spectrophotometry on the above apparatus: for fat 3.48 microns, for proteins 6.46 microns and for lactose 9.60 microns. Bulk milk samples used for the tests were untreated or treated with potassium dichromate, bronopol, sodium azide and Milkofix at the temperatures of storage in darkness 20 degrees C and 4 degrees C. The differences against the reference values (measured on the first day) were determined and evaluated in milk composition and characteristics as arising during milk storage. These differences were used in form of either cumulative means of differences (Figs. 1 to 5) or individual differences (Fig. 8). In the first part significant correlation coefficients (P less than 0.001) were calculated for the relationship between the variations of lactose content and the fat and protein contents: r = -0.59 and/or -0.73 (Figs. 6 and 7). This suggests that the decrease in the lactose content by 0.10% recorded by the infrared analysis and caused by lactose decomposition is accompanied by a "seeming" increase in the fat and protein content by about 0.04%. In the second part the correlation coefficients for the fat and protein contents r = -0.96 and -0.96 (P less than 0.001; Figs. 9 and 10; Tab. II) were calculated on the basis of an observation of the lactose decrease in an untreated milk sample (20 degrees C for 28 hours). These coefficients are somewhat different from the preceding ones; this is due to the lower homogeneity of the first set where the milk samples were treated in a different way, but the coefficients confirm the same conclusions. The values of the correlation coefficients for the dependence between the development of the acquired titratable acidity (SH) and the variations of fat (F), protein (P) and lactose (L) contents were as follows: r = 0.95; 0.95; -0.99 (P less than 0.001; Figs. 12, 13; Tab. II). Thus the above-mentioned "seeming" increase in the F and P contents can be explained to the extent of 92.2% from the decrease in the L content, which also causes the increase in titratable acidity to the extent of 98.0%.(ABSTRACT TRUNCATED AT 400 WORDS)     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

结球莴苣“烧边”成因及其调控措施的研究进展

结球莴苣烧边病是生理性病害，严重影响其品质，制约生产。国内外多数研究者认为钙缺乏导致结球莴苣烧边病的发生，但是现在许多研究表明其他因素也能导致烧边病的出现，例如环境因素、生长速率、内源激素等，所以研究烧边病发生的机制及其与环境因素的关系是防御结球莴苣烧边病发生的关键。     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001     

Induction of an antioxidant enzyme system and other oxidative stress markers associated with compatible and incompatible interactions between chickpea (Cicer arietinum L.) and Fusarium oxysporum f. sp.ciceris

To ascertain if active oxygen species play a role in fusarium wilt of chickpea caused by Fusarium oxysporum f. sp. ciceris, the degree of lipid peroxidation (malondialdehyde formation) and the activity levels of diamine oxidase (DAO), an apoplastic H₂O₂-forming oxidase, and several antioxidant enzymes, namely ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guaiacol-dependent peroxidase (GPX) and superoxide dismutase (SOD), were determined spectrophotometrically in roots and stems of ‘WR315’ (resistant) and ‘JG62’ (susceptible) chickpea cultivars inoculated with the highly virulent race 5 of the pathogen. Moreover, APX, CAT, GPX and SOD were also analysed in roots and stems by gel electrophoresis and activity staining; and the protein levels of APX and SOD in roots were determined by Western blotting. In roots, infection by the pathogen increased lipid peroxidation and CAT and SOD activities, although such responses occurred earlier in the incompatible compared with the compatible interactions. APX, GPX and GR activities were also increased in infected roots, but only in the compatible interaction. In stems, infection by the pathogen increased lipid peroxidation and APX, CAT, SOD and GPX activities only in the compatible interaction, and DAO activity only in the incompatible one. In general, electrophoregrams agreed with the activity levels determined spectrophotometrically and did not reveal any differences in isoenzyme patterns between cultivars or between infected and non-infected plants. Further, Western blots revealed an increase in the root protein levels of APX in the compatible interaction and in those of SOD in both compatible and incompatible interactions. In conclusion, whereas enhanced DAO activity in stems, and earlier increases in lipid peroxidation and CAT and SOD activities in roots, can be associated with resistance to fusarium wilt in chickpea, the induction of the latter three parameters in roots and stems along with that of APX, GR (only in roots) and GPX (only in stems) activities are rather more associated with the establishment of the compatible interaction.     

细胞骨架解聚药物对小麦与叶锈菌互作诱发的细胞过敏性反应的影响

 小麦(Triticum aestivum)品种洛夫林10和叶锈菌小种366组成不亲和组合,小麦叶片发生过敏性坏死反应(HR)是小麦抵抗叶锈菌侵染的重要因素。在接种前给小麦叶片分别预注射微管解聚药物磺草硝(oryzalin)和微丝解聚药物细胞松弛素D (cytochalasin D,CD),结果表明2种药物注射使得寄主因叶锈菌侵染诱导的细胞过敏性坏死数目明显减少,并且注射药物的浓度越大,寄主细胞发生HR的数量越少。说明肌动蛋白和微管蛋白的聚合状态是诱发小麦叶片发生HR防卫反应所必需的,细胞骨架在小麦抵抗叶锈菌侵染过程中可能起着重要作用。     

红肉脐橙果肉中主要色素的定性及色素含量的变化

 运用薄层层析法及二极管阵列检测器与高效液相色谱联用的方法, 确定红肉脐橙果肉中含有番茄红素和β- 胡萝卜素两种主要色素, 它们在液相色谱图中占总峰面积(计溶剂峰) 的73. 57 %～77. 78 % , 番茄红素占55. 36 %～58. 49 % , β- 胡萝卜素占18. 21 %～19. 93 %。利用反相高效液相色谱仪以及番茄红素和β- 胡萝卜素标样制作标准曲线, 测定果实发育及室温贮存过程中色素的变化动态, 结果表明, 该脐橙果肉中的番茄红素在成熟期含量最高, 达109. 67μg·g^-1DM, β- 胡萝卜素此时也达到最高值(15. 52μg·g^-1DM) ; 番茄红素的含量在采收后基本呈下降趋势(但采后30 d 有显著升高) , 说明果肉中存在类胡萝卜素采后合成的现象; 贮存4 个月后, 番茄红素含量急剧降低至最高时的1/ 10。     

红肉脐橙(Citrus sinensis L.)果肉中特征色素提取方法探索

红肉脐橙是目前为止惟一果肉呈粉红色或红色的脐橙,其呈色色素尚未见报道。为深入研究其果肉中特征色素的构成及在果实生长发育和贮存过程中色素含量的变化,本研究在对该色素初步定性的基础上对提取方法进行了探索。在分析红肉脐橙果肉中色素萃取液的紫外可见吸收光谱时发现,该色素有类胡萝卜素的特征吸收峰,将萃取液进行薄层层析则进一步证实其内含有番茄红素。在色素提取的前处理方式上,本研究认为冷冻干燥样品的浸提效果较真空干燥的样品稳定(变异系数小),且浸提率相差无几,所以选择冷冻干燥作为合适的样品前处理方式。色素浸提条件的正交实验结果表明,最佳浸提条件为50℃、1.5h内浸提4次,浸提的料液比为1:60。此外,浸提温度、浸提时间、浸提料液比和浸提次数对浸提率都有显著影响,影响最大的因素是浸提温度,最佳温度为50℃,随温度降低浸提率显著降低。其次是侵提时间的长短,最佳浸提时间为1.5h,延长或缩短0.5h都会使浸提率下降。浸提次数对浸提率的影响是最小的。     

柑橘类果实汁胞的红色现象及其呈色色素

柑橘类果实中不乏汁胞为红色类型者,如红肉葡萄柚(Citrus paradisii Macf.)的粉红色是番茄红素所致,血橙(C.sinensis L.)的血红色来源于花青素,此外,还有红色的红江橙(C.sinensis L.)、橙红色的元红(C.reticulata Blan-co)和紫红色的五布红心柚(C.grandis L.)等,它们因色泽鲜艳而具备特殊的市场潜力,但呈色色素尚未见报道。在符合审美观的同时,红色汁胞品种还具有特殊的营养价值,如类黄酮、类胡萝卜素等色素富集可软化血管、抗癌、抗氧化及消除体内自由基等。综合有关文献,首次将国内外主要红色汁胞柑橘品种依宽皮柑橘、甜橙、柚、葡萄柚及杂柑等分类进行综述,分析可能的呈色色素,阐明导致红色性状产生的两类色素—花青素、类胡萝卜素生物合成途径