油茶籽中美拉德反应产物的抗氧化性及其含量分析

[目的]研究前处理对油茶籽的抗氧化性影响。[方法]采用红外加热对油茶籽进行不同时间与温度的处理,并测定处理前后油茶籽和油茶籽仁中美拉德反应产物含量及其抗氧化性。[结果]研究发现随加热时间的延长,美拉德反应产物5-羟甲基糠醛、丙酮醛、乙二醛、3-脱氧奥苏糖的生成量逐渐增多;在150℃下反应120 min后,油茶籽中5-羟甲基糠醛、丙酮醛、乙二醛、3-脱氧奥苏糖的含量分别为10.929、34.620、11.054和36.627μg·g~(-1),油茶籽仁中其含量分别为16.950、55.217、20.216和72.390μg·g~(-1),后者分别是前者的1.55、1.59、1.8和1.97倍,说明美拉德反应主要在油茶籽仁中发生。对美拉德反应产物氧化活性的测试数据表明,油茶籽中的美拉德反应产物均具有抗氧化活性,在150℃下反应120 min时,油茶籽仁和油茶籽清除DPPH自由基与抗氧化活性均达到最高,前者分别为99.18%和81.47%,后者分别为97.44%和74.97%。[结论]比较发生美拉德反应的油茶籽仁与油茶籽的DPPH清除率与抗氧化活性发现,油茶籽仁抗氧化活性强于油茶籽抗氧化活性。     

小麦种子规模化DNA提取及检测体系的优化

以小麦种子分子检测农业行业标准(NY/T 2859-2015)为基础,开发小麦种子快速、规模化提取DNA方法,优化反应体系中各组分含量,为小麦真实性快速执法提供技术支撑。通过对SDS、CTAB、高盐低p H、快速提取和试剂盒5种方法提取的DNA质量、浓度和PCR扩增效果进行比较,发现改进的高盐低p H方法提取种子的DNA质量、浓度能够满足小麦真实性鉴定中42对SSR引物重复鉴定的需求,是利用96孔深孔板和自动化移液工作站规模化、高通量提取DNA的较优方法。进一步优化结果显示该方法在65℃温浴条件下比室温的浓度高出25%,但两种温度条件下提取的DNA质量及浓度均能够满足品种鉴定的需求;沉淀时用0.5倍提取液体积的预冷异丙醇沉淀浓度最高,用室温的异丙醇沉淀的最佳体积是提取液体积的0.6倍。对标准中42对引物的最佳引物浓度和模板浓度均进行了优化,综合所有42对引物的优化结果发现,反应体系为20μL时,引物终浓度为0.437 5μmol/L,模板终浓度为10 ng/μL时扩增效率相对较高,扩增产物稳定,能够满足多重电泳的需求。     

Nutrient release pattern of organic and inorganic resources used in cassava production (Manihot esculenta Crantz)

The major constraint in the promotion of organic farming, a safe and sustainable alternative, is lack of availability of quality organic resources in sufficient quantities. The quality of resources can be ascertained by monitoring its nutrient release pattern, which has not yet been attempted. A controlled condition pot experiment was conducted at ICAR-Central Tuber Crops Research Institute, Thiruvananthapuram, India, in completely randomized design to study the nutrient dynamics of various organic and inorganic resources, commonly used in cassava production, at monthly intervals up to six months. The release of almost all nutrients and activity of soil enzymes were higher at the middle (3 or 4?months) of the incubation period. The pH showed an increasing trend and electrical conductivity, organic C and Fe content declined from initial. Averaging over stages, organic practice favored the activity of soil enzymes and release of almost all nutrients over conventional system significantly, except N.     

马铃薯卷叶病毒PLRV RT-LAMP检测方法优化

马铃薯卷叶病毒Potato leafroll virus(PLRV)是目前严重影响马铃薯产量与品质的主要病毒之一,给马铃薯产业造成巨大损失。本研究采用环介导等温核酸扩增(loop-mediated isothermal amplification, LAMP)技术建立PLRV的RT-LAMP检测方法。采取单因素变化试验,对RT-LAMP反应体系中多个因素包括引物组合、温度条件及Mg~(2+)、betaine、Bst 3.0 DNA聚合酶、dNTPs、UNG、SYBR GreenⅠ和引物组合的浓度进行一系列试验和优化。采用RT-PCR检测方法进行平行比对试验,对优化后的RT-LAMP反应体系进行了验证。结果表明,最佳引物组合为P3,最适反应温度62℃,25μL反应体系中,Mg~(2+)、betaine、Bst 3.0 DNA聚合酶和UNG的最佳终浓度分别为4 mmol/L、0 mmol/L、0.64 U/μL和0.08 U/μL,dNTPs的最佳用量为1μL(dATP、dGTP、dCTP各0.4 mmol/L,dUTP 1.2 mmol/L),SYBR GreenⅠ(20×)的最佳用量1μL,primer mix的最佳用量2.5μL(PLRV-FIP/BIP、PLRV-F3/B3和PLRV-LF/LB的浓度分别为0.8、0.2μmol/L和0.6μmol/L),RNA模板1μL(2 ng/μL),加DEPC-H_2O至25μL,反应时间50 min。优化后的RT-LAMP检测结果与RT-PCR一致,且可视化判读结果。因此,建立的PLRV RT-LAMP检测方法为进一步开发RT-LAMP检测试剂盒及其实际应用奠定了基础。     

根皮苷氧化物POP2的酶促反应体系及其结构表征

【目的】根皮苷氧化物POP2是苹果汁加工过程中由酶促氧化生成的一种多酚氧化物,对果汁的褐变和劣变有着重要影响。通过多酚氧化酶氧化根皮苷反应体系的建立,POP2的分离纯化与结构表征,探讨根皮苷氧化物POP2合成条件与反应历程,为苹果汁中POP2的检测分析,及弄清其对果汁褐变和劣变的影响奠定基础。【方法】通过p H、温度、底物浓度3个单因素试验建立根皮苷酶促氧化体系的基础上,研究乙醇溶液对酶促反应的促进作用和反应历程,通过高效液相色谱法检测根皮苷酶促氧化过程中根皮苷、中间产物及POP2含量的变化,通过乙酸乙酯萃取、无水乙醇除盐、重结晶等方法对氧化产物POP2进行分离纯化,并采用质谱(ESI-MS)和红外光谱(IR)对POP2进行结构表征。【结果】根皮苷酶促氧化反应的最佳条件为p H 6.5、35℃、底物浓度3 mg·m L~(-1),添加不同浓度的无水乙醇均能显著提高根皮苷酶促氧化速率,当添加浓度为20%时,可提高反应转化率,最大为2.4倍;根皮苷反应液随时间的变化颜色也发生改变,反应刚开始溶液颜色呈微黄色,随着反应的进行颜色逐渐从亮黄色变为橙黄色,最终产物的颜色为橙红色;高效液相色谱法分析根皮苷酶促氧化反应历程发现,当底物浓度为3 mg·m L~(-1),反应48 h时,根皮苷的转化率最大为90.2%,中间产物X1和POP1也基本接近痕量,POP2的积累量基本不再发生变化;通过乙酸乙酯萃取可以将反应液中残留的根皮苷除去,用无水乙醇除盐并经甲醇重结晶后,获得纯度为96.8%的POP2;经正离子模式ESI-MS分析,POP2的m/z为467,分子式为C_(21)H_(22)O_(12);IR结果发现POP2具有特征性的醚、氧化羧基、酮基及苯环官能团,与推测的根皮苷氧化物结构式吻合。【结论】根皮苷酶促氧化体系中的主要氧化物为POP2,可以根据POP2的含量高低建立苹果汁褐变和劣变的标志性检测指标。     

Production of Antioxidative Maillard Reaction Products from Gelatin Hydrolysate of Unicorn Leatherjacket Skin

Antioxidative activities of Maillard reaction products (MRPs) derived from gelatin hydrolysate (GH) of unicorn leatherjacket skin as affected by types of saccharides, GH to saccharide ratio, incubation temperatures, relative humidity (RH), and times were investigated. MRPs prepared using mixture of GH and galactose at 2:1 (w/w) ratio showed the highest antioxidative activity. At different incubation temperatures (50–70°C) and RHs (55–75%), optimal condition for preparing antioxidative MRPs included heating GH and galactose (2:1) powder at 70°C and 55% RH for 36 h. Electrophoresis and Fourier transform infrared spectroscopy revealed the formation of high molecular weight polymers, and the advanced glycation took place in MRPs.     

A comparison of changes in stable isotope ratios in the epidermal mucus and muscle tissue of slow‐growing adult catfish

Although stable isotope analysis is a powerful tool for determining diet, migration patterns and the structure of food webs in aquatic systems, the slow response of isotopic ratios in the widely used muscle tissue often hampers this approach, particularly in slow‐growing or adult fishes. We conducted a diet‐switch experiment to compare the changes in the stable carbon isotope ratio (δ¹³C values) in the epidermal mucus and muscle tissue of five‐year‐old catfish (Silurus asotus). The isotope ratios in noninvasively sampled mucus changed more rapidly than those in the muscle tissue. As isotopic change in mucus was relatively rapid, this technique can be used over a finer timescale than traditional isotopic analyses using the muscle tissue. The isotopic change half‐life in our experimental condition was 200 days, which would not be short enough for some research purposes. Examining mucus along with the muscle would enable food habits of slow‐growing fishes to be determined over different timescales. However, the rate of isotopic change in the mucus was negatively affected by the size of fish and was slower than the previously reported rates in juvenile steelhead. These findings suggest that mucus turnover rates need to be determined prior to the field data interpretation.     

Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.