凡纳滨对虾α-淀粉酶基因的SNPs检测

利用PCR产物直接测序法,对凡纳滨对虾SPR和SPF 2个品种共40个样本的α-淀粉酶基因(GenBank序列号:AJ133526)的单核苷酸多态性(SNP)进行了研究。找到9个SNPs,分别为:C127T、A138G、T951C、T1083C、C1457T、A2147C、A2226G、A2297G和T2306C。其中1个SNP是第4外显子中的错义突变,2个SNPs分别是第5和第7外显子中的同义突变,6个SNPs是内含子中的突变。这些结果可为凡纳滨对虾的研究提供遗传学依据。     

小麦抗穗发芽机理研究现状

为了及时了解小麦抗穗发芽机制,加快我国小麦抗穗发芽品种选育进程。以影响穗发芽的主要因素α-淀粉酶为切入点,对影响α-淀粉酶活性的调控因子α-淀粉酶抑制剂和硫氧还蛋白h在小麦中的分布、作用性质、生物学功能及抗穗发芽解决途径进行了介绍。并结合自己的研究工作,认为利用基因工程技术导入反义Trxs基因是解决穗发芽的一条行之有效的途径。     

Chemical Constituents of Lagenaria siceraria Mesocarp and Its Xanthine Oxidase and Alpha-Amylase Inhibitory Activities

The present study was conducted to identify natural products in the bioactive fractions of Lagenaria siceraria mesocarp. Column chromatography and gas chromatography-mass spectrometry (GC-MS) was used for separation and identification of phytochemicals. Methanolic extract and its fractions displayed alpha-amylase and xanthine oxidase inhibitory activities comparable to standard drugs. GC-MS analysis of the fractions and column eluates revealed the presence of a variety of chemical compounds, including 4-(methoxymethyl)phenol, 2,2?-methylenebis{6(1,1-dimethylethyl)-4-ethyl}phenol, methyl 2-hydroxy-3-phenylpropanoate, oxacyclododecane-2,8-dione, N-(2-(4-hydroxyphenyl)ethyl] acetamide, 2,3-Epoxycarane, 4-(propan-2-yl)benzaldehyde, octadec-1-ene, 4a-methyl-1-methylidene-1,2,3,4,4a,9,10,10a-octahydrophenanthrene, 2,6-di(propan-2-yl)naphthalene, methyl 3-(4-hydroxy-3,5-dimethylphenyl)propanoate, 4,8a-dimethyl-6-(prop-1-en-2-yl)-3,5,6,7,8,8a-hexahydronaphthalen-2(1H)-one 1,3-dihydro-2-benzofuran, methyl N-hydroxybenzenecarboximidoate, linoleic acid, and palmitic acid. Many of these compounds are being reported for the first time from this plant, the presence of which may explain, at least partly, its medicinal efficacy. The fruit possesses significant anti-enzymatic properties against alpha-amylase and xanthine oxidase, and the most active compounds of mesocarp of the fruit appear in ethyl acetate fraction of its methanolic extract.     

乳酸菌发酵驼乳工艺优化及抑制α-淀粉酶和α-葡萄糖苷酶活性的研究

[目的]以实验室前期从驼乳中分离得到的乳明串珠菌、副干酪乳杆菌为对象,并通过体外实验对发酵驼乳抑制α-淀粉酶和α-葡萄糖苷酶活性以及抗氧化能力进行测定。[方法]以α-淀粉酶抑制率和α-葡萄糖苷酶抑制率为考察指标,在单因素试验的基础上,采用正交试验优化发酵驼乳的参数条件。测定在不同储藏时间下试验组乳明串珠菌、副干酪乳杆菌发酵驼乳、对照组乳明串珠菌、副干酪乳杆菌发酵牛乳和嗜热链球菌、保加利亚乳杆菌发酵驼乳pH值、酸度、活菌数、α-淀粉酶抑制率、α-葡萄糖苷酶抑制率、羟自由基和超氧阴离子自由基清除率的变化。[结果]驼乳发酵的最佳工艺条件为乳明串珠菌∶副干酪乳杆菌比例2∶3、接种量2%、发酵温度39 ℃,发酵时间24 h,此条件下测得对α-淀粉酶和α-葡萄糖苷酶的抑制率分别达到87.20%和20.85%,与未优化前相比,α-淀粉酶的抑制率提高14.10%,α-葡萄糖苷酶抑制率提高4.4%。在4 ℃储藏期间发酵驼乳α-淀粉酶抑制率在第6d时最高为90.39%,对α-葡萄糖苷酶的抑制率最高为36.32%,抑制率显著高于对照组。各组对羟自由基的清除率达到70%以上,最高为94.91%,超氧阴离子自由基清除率最高达到78.10%,整个测定过程中试验组的抗氧化活性均显著高于对照组。[结论]经乳酸菌发酵驼乳抑制α-淀粉酶和α-葡萄糖苷酶活性提高,在储藏期间抗氧化活性显著高于对照组（P<0.05）。     

Pyramiding genes affecting sprouting resistance in Rye by means of marker assisted selection

The 541 × Ot1-3 intercross population and bulked segregant analysis (BSA) were used to search for molecular markers linked to genes underlying sprouting and alpha-amylase activity. Six RAPD markers showing association with studied traits were tested for their potential effectiveness in selecting sprouting resistant genotypes. It was shown that although individual effects of markers were not high, their accumulation in one genotype gives substantial increase in sprouting resistance.     

鹰嘴豆α-淀粉酶抑制剂基因CL-AI的原核表达、纯化及生物学特性

为研究鹰嘴豆种子中新贮藏蛋白类型α-淀粉酶抑制剂CL-AI的抑制活性、生物活性以及三级结构,本研究利用前期克隆的α-淀粉酶抑制剂基因CL-AI及其2个亚基CL-AI-α、CL-AI-β分别构建原核表达载体p E-SUMO3,经诱导表达获得了CL-AI、CL-AI-α的可溶性融合蛋白和CL-AI-β包涵体融合蛋白,其中CL-AI-β通过构建含不同标签的表达载体,最终实现了可溶性表达。人唾液淀粉酶(HSA)抑制活性试验结果表明,CL-AI,CL-AI-α,CL-AI-β3个蛋白对HAS都有一定的抑制作用。发芽试验表明,CL-AI蛋白对小麦、玉米、水稻种子发芽过程中的发芽势、根长和芽长影响显著,与加入无菌水的处理相比,种子的简化活力指数显著下降。本研究结果为CL-AI蛋白的后续研究与应用奠定了基础。     

Effects of cold temperature on starch molecular structure and gelatinization of late-maturity alpha-amylase affected wheat

The synthesis of late-maturity alpha-amylase (LMA) occurs when wheat experiences a cold-temperature ‘shock’ during the post-anthesis, grain-filling period. This markedly increases the high isoelectric point (pI) alpha-amylase in wheat grains in susceptible varieties. The affected grain has low Falling Numbers (FN) and is rejected at receival point or downgraded to feed grade, as low FN is associated with inferior end-product quality. However, several studies have reported the lack of correlation between low FN-LMA and end-product quality. Here, we characterize, for the first time, starch molecular structure and gelatinization properties of cold-treated wheat grains; starch structure has significant influence on flour functionality and end-product quality. Results show that the cold-treatment during post-anthesis has minimal effect on starch structure. While there was a small decrease in the gelatinization temperature for every wholemeal flour samples from cold-treated wheat grains, this is unlikely to cause any undesirable effect on end-product. The present findings suggest that the supposition that LMA is a major contributor to inferior end-product quality should be reconsidered.     

盐度和胁迫时间对九孔鲍α-淀粉酶基因表达量的联合效应分析

采用中心复合设计和响应曲面法研究了盐度和胁迫时间对九孔鲍(Haliotis diversicolor supertexta)?-淀粉酶基因表达量影响的联合效应,旨在为九孔鲍盐度驯化及海区推广提供指导。实验设定盐度范围为22~40,时间范围为0~72 h,建立了盐度、胁迫时间与?-淀粉酶基因表达量之间关系的定量模型,并通过分析明确了盐度和胁迫时间的最优组合。结果表明,盐度对九孔鲍?-淀粉酶基因表达量影响的一次效应不显著(P0.05),而二次效应极显著(P0.01),说明盐度与九孔鲍?-淀粉酶基因表达量之间为非线性关系。随盐度升高,淀粉酶基因表达量呈先上升后下降的变化趋势,当盐度为31时淀粉酶基因表达量最大,此时九孔鲍消化能力最强。胁迫时间对淀粉酶基因表达量影响的一次效应显著(P0.05),二次效应不显著(P0.05),表明胁迫时间与九孔鲍?-淀粉酶基因表达量之间呈线性关系,即随着胁迫时间的延长,淀粉酶基因从开始低水平表达逐渐升高并恢复至正常表达水平。盐度和胁迫时间之间不存在交互作用。对实验结果建立模型可知,?-淀粉酶基因表达回归模型达极显著水平(P0.05),且失拟项不显著(P0.05),表明实验拟合出的方程有效。回归方程的决定系数为80.05%,校正系数65.80%,预测系数50.18%,表明方程拟合度较好,模型选择较为恰当,可以为不同盐度和胁迫时间诱导下的淀粉酶基因表达量变化提供参考依据。     

小麦抗穗发芽机理研究现状

为了及时了解小麦抗穗发芽机制，加快我国小麦抗穗发芽品种选育进程。以影响穗发芽的主要因素α-淀粉酶为切入点，对影响α-淀粉酶活性的调控因子α-淀粉酶抑制剂和硫氧还蛋白h在小麦中的分布、作用性质、生物学功能及抗穗发芽解决途径进行了介绍。并结合自己的研究工作，认为利用基因工程技术导入反义Trxs基因是解决穗发芽的一条行之有效的途径。