玉米品质基因聚合对子粒氨基酸组分及醇溶蛋白的影响

以玉米o2、sh2、su1、wx这4个品质基因聚合的单基因、双基因、三基因近等基因系为研究对象,对子粒氨基酸组分与醇溶蛋白SDS-PAGE带谱进行检测分析。结果表明,与郑58相比较,su1、wx、su1wx氨基酸组分变化较小,o2、sh2单基因以及含有o2、sh2的双基因、三基因近等基因系中氨基酸组分变化较大,o2sh2、o2su1、o2sh2su1赖氨酸含量较高。SDS-PAGE显示,o2与sh2、su1聚合导致19 kD和22 kDα-醇溶蛋白合成受到显著抑制。     

玉米子粒粉质突变体3901l的图位克隆

粉质胚乳突变体3901l来源于美国种质中心,该突变体胚乳呈现完全不透明表型。生化分析发现,该突变体α-和β-醇溶蛋白剧烈下降,同时伴随着非醇溶蛋白的大幅度升高。通过玉米SNP3072芯片进行基因分型分析发现,3901l基因定位于7号染色体约33 M的区间。通过与o2突变体进行等位测试,确认3901l是o2的等位突变体。进一步研究表明,该突变体的O2转录本缺失了一个61-bp的外显子,造成开放阅读框移码和翻译提前终止。利用O2特异抗体检测子粒总蛋白,发现突变体中O2蛋白完全缺失,不能检测到提前终止的O2蛋白,说明错误翻译的O2蛋白被机体快速识别并降解。     

Physical properties of zein films containing salicylic acid and acetyl salicylic acid

Zein films containing salicylic acid (SA) and acetyl salicylic acid (ASA) between 2 and 10% (initial zein weight basis) with or without glycerol were evaluated for structure, mechanical and dissolution properties. The random coils, α helices and β sheets mainly governed the secondary structure of zein, depending on glycerol and level of model molecules. Adding ASA resulted in an increase in α helices whereas β sheets increased at the expense of α helices when SA was used. Including SA or ASA decreased the tensile strength and the stiffness of films containing glycerol indicating the synergistic effect of SA and ASA. The strain at failure decreased with increasing content of SA but increased with increasing level of ASA. The dissolution properties were glycerol and drug dependent. ASA release in comparison to SA was quite low. The release was only observed above 10% ASA whereas it was detected in all films containing SA. The possible interactions between active components and proteins are discussed together with their implications on the physical properties of zein films.     

非常规饲料原料饲喂肉牛育肥的试验研究

[目的]本研究旨在分析全混合日粮中添加苹果渣、玉米纤维及玉米酒精糟三种非常规饲料原料对育肥肉牛采食量、增重效果、产肉性能及经济效益的影响。[方法]选用体重（455±10）kg、健康年龄相近的西门塔尔杂交品种育肥牛14头,随机分为2组,每组7头,设对照组和试验组。分别饲喂未添加非常规饲料原料和添加非常规饲料原料的饲料。[结果]显示：试验组较对照组日增重、屠宰率、净肉率、胴体产肉率、眼肌面肌分别提高了1.1%、2.1%、2.1%、3.3 cm2。出售活牛试验组头均盈利比对照组多出205.2元、牛副产品头均盈利值多出586.8元。[结论]表明在肉牛育肥中,应用推广苹果渣、玉米酒精糟及玉米纤维等非常规饲料原料,具有扩大饲料来源、降低饲养成本等优点,能进一步提高肉牛的增重速度与产肉性能,增加养殖的经济效益。     

玉米醇溶蛋白基因Zein启动子克隆与序列分析

启动子是细胞内控制基因转录与表达的时空特异性的重要因子。玉米醇溶蛋白基因Zein启动子控制Zein基因在胚乳中特异性表达,分离Zein启动子将为外源基因在胚乳中特异性表达等遗传操作提供基础。以玉米自交系鲁2548基因组DNA为模版,PCR扩增得到401 bp的15 kuβ-Zein启动子,PCR产物T-A克隆并测序。序列分析结果显示,作者克隆的Zein启动子序列与NCBI已报道的15 ku-Zein基因(GenBank M72708)启动子序列同源性为96%。启动子功能预测及顺式元件分析表明,所克隆的启动子序列可能具有胚乳特异性启动子功能。有关功能验证试验正在开展之中。本研究为外源基因胚乳特异性表达基因工程奠定了基础。     

Characterization of zein modified with a mild cross-linking agent

Zein, a predominant corn protein, is an alcohol-soluble protein extracted from corn and is an excellent film former. The characteristic brittleness of zein diminishes its usefulness as a film. It is well known that zein has a propensity for forming aggregates in solution. When zein molecules were cross-linked with 1-[3-dimethylaminopropyl]-3-ethyl-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), it was found that the film-forming property was improved and the aggregation phenomenon in solution was suppressed. At the air/water interface, native zein forms brittle film with rough surface, whereas cross-linked zein forms rigid film with very smooth and even surface. Tensile strengths of the films were shown to be greatly increased by cross-linking. Objectives of this study are to determine the cross-linking mechanism of zein, the optimum reaction conditions, characteristics of the reaction product, and mechanical properties of cross-linked zein film. Through viscosity and dynamic light scattering, the cross-linking reaction was monitored. Optimum amount of EDC and NHS was determined to be 30 mg each per gram of zein. The cross-linking of zein with EDC and NHS seemed to be self-terminating because the cross-linking reaction did not proceed to precipitation.     

玉米opaque2突变体子粒表型及营养成分分析

通过回交转育构建o2突变型的近等基因系CAL58/o2,对其子粒进行扫描电镜和透射电镜观察,发现与CAL58相比,CAL58/o2中淀粉颗粒排列松散,形状更加规则,CAL58/o2子粒中蛋白体体积变小。分析子粒的营养成分,CAL58/o2赖氨酸含量极显著提高了55.88%,氨基酸总量显著减少9.79%,粗蛋白显著减少8.86%,粗脂肪显著增加30.96%,粗淀粉显著增加5.87%。同时,提取成熟子粒醇溶蛋白进行比较,CAL58/o2子粒的α、β-醇溶蛋白含量明显降低,γ-醇溶蛋白含量增加。     

Removal of surface lipids improves the functionality of commercial zein in viscoelastic zein-starch dough for gluten-free breadmaking

Maize prolamin (zein), together with starch, hydroxypropyl methylcellulose, sugar, salt, yeast and water can form wheat-like cohesive, extensible, viscoelastic dough when mixed above room temperature (e.g. 40 °C). This dough is capable of holding gas. However, it is excessively extensible, and when used for hearth-type rolls, it tends to become flat. Bench-scale defatting of zein with chloroform at room temperature significantly (P < 0.05) improved specific volume (4.5 ml/g vs. 3.3 ml/g) and shape of the rolls (width-to-height 2.0 vs. 3.9). The total lipid content determined by accelerated solvent extraction (100 °C, 69 bar, chloroform), however, only decreased from 8.0 to 6.6% due to this bench-scale defatting. Staining experiments with Naphthol Blue Black suggested that bench-scale defatting removed surface lipids from the zein particles, and thus facilitated their aggregation. Aggregation experiments with zein and water at 40 °C, and laser scanning confocal microscopy with zein-starch dough confirmed that zein particles aggregated more easily when surface-defatted. Dynamic oscillatory temperature sweeps demonstrated that surface-defatting lowered the temperature at which protein cross-linking occurred by 2 °C. This research can help to produce superior gluten-free bread and could also possibly contribute to the better understanding of wheat dough.     

Impact of different isolation procedures on the functionality of zein and kafirin

Commercial corn prolamin (zein) aggregates in water at elevated temperatures into an extensible, viscoelastic gluten-like substance. This specific functionality of zein can be used in the production of gluten-free bread from true dough systems and not from batters. The present study examined laboratory-scale isolation of such functional zein from dry milled corn. RP-HPLC indicated that successful isolation procedures resulted in relatively pure α-zeins, with a maximum ratio of (β + γ)/α-zeins of about 10%. In the present study, such functional zeins were obtained by using 70% ethanol as the extractant, without added alkali or reducing agent in the main extraction step. In contrast, films could be cast from a wider range of zein isolates, also with higher ratios of (β + γ)/α-zeins. Isolation of the analogous prolamin (kafirin) from dry milled sorghum required a more hydrophobic extractant such as 83% isopropanol to achieve partial functionality. Such kafirin was able to aggregate in warm water, preferably when a reducing agent was added; however, it quickly became firm and lost its extensibility. The present study suggests that hydrophobic interactions rather than disulfide bonds are the key to gluten-like functionality of zein and kafirin.     

Melt-processed blends of zein with polyvinylpyrrolidone

Melt-processed blends of zein and polyvinylpyrrolidone (PVP) of varying molecular weights (55K, 360K and 1.3M) were compared based on mechanical and thermal properties. Generally, all samples stored at 50% RH exhibited a slight improvement in tensile strength, with the PVP360K samples showing the greatest improvement. At the higher levels of PVP, samples stored at 70% RH showed a decrease in tensile strength. Elongation was also more significantly impacted at higher humidity, with the higher levels of PVP causing greater elongation increases. Differential scanning calorimetry data for the blends showed single T_g values intermediate between the zein and PVP controls. Kinetic thermogravimetric data suggested a multi-step degradation interaction for the zein/PVP blends. Scanning electron microscope imaging of compression molded samples showed homogeneous surface contours for even the 20% PVP1.3M blend. Melt-processed blends of zein with polyvinylpyrrolidone of various molecular weights appear to be compatible. This work represents the first melt-processed blend of zein with PVP to generate a compatible blend.