Antimicrobial effects of black rice extract on Helicobacter pylori infection in Mongolian gerbil

It has been reported that cyanidin 3-O-glucoside (C3G) is an inhibitor of Helicobacter pylori toxin secretion. C3G is classified as an anthocyanin and is a major component of black rice extract (BRE). The present study aimed to identify a new functional food material to prevent H. pylori infection in Mongolian gerbil model. Toxicity in the liver and kidney were not detected after BRE administration (10 or 50 mg/kg). BRE treatment reduced bacterial colonization in animal gastric tissue, as well as infection signs as observed on the analysis of the hematological data. It was also found that the relative mRNA levels of the inflammatory cytokines were reduced in BRE-treated groups. These findings suggest that BRE acts as a potent inhibitor of H. pylori infection and pathogenesis in Mongolian gerbils. We propose that BRE may be used to manage gastroduodenal diseases caused by H. pylori infection.     

Wx-1基因变异和优质HMW-GS组合对中筋小麦品质的影响

高分子量谷蛋白亚基(HMW-GS)和Wx蛋白是两个影响小麦理化品质和制成品品质的重要因素。为研究HMW-GS和Wx基因变异对小麦理化品质、面条感官评分和质构特性的影响,以镇麦9号和扬糯麦1号为亲本的2个高代系为试验材料,研究了优质HMW-GS 5+10及Wx基因缺失对小麦理化品质及面条加工品质的效应。结果表明,品系1与品系2的湿面筋含量和SDS沉淀值与镇麦9号相近,但显著好于扬糯麦1号。相较于镇麦9号,品系1与品系2的直链淀粉含量分别降低了1.5%和2.5%,其峰值黏度和稀懈值均显著高于镇麦9号,膨胀势也高于镇麦9号。5+10亚基显著提高了面条质构参数中的硬度和咀嚼性,而Wx基因缺失显著提高了面条的软硬度评分和光滑性评分。综合来看,具有5+10亚基和 Wx-D1缺失型的品系2具有较高的SDS沉淀值、面团形成时间和稳定时间、较低的直链淀粉含量、较高的峰值黏度、稀懈值和膨胀势,蛋白质和淀粉综合品质表现较好。品系2面条总评分最高,显著高于镇麦9号和扬糯麦1号,其主要体现在适中的面条软硬度和较好的光滑性。推测Wx基因缺失和优质HMW-GS聚合可显著提高小麦淀粉品质和蛋白质品质,有效改善面条的蒸煮品质和感官评分。     

晋豆24~(60)Coγ射线诱变后代种子贮藏蛋白的研究

选用晋豆24及其60Coγ射线辐照诱变后代为试验材料,利用SDS-PAGE聚丙烯酰胺凝胶电泳研究了诱变后代大豆贮藏蛋白亚基11S和7S的组分含量、亚基组成和各亚基的含量及其变异规律。结果表明:大豆贮藏蛋白种质资源中,同一品种的不同亚基和不同品种的同一亚基间均有较大变异;11S球蛋白与7S球蛋白呈极显著负相关,与11S/7S比值呈极显著正相关;聚类分析根据11S/7S比值将诱变后代群体分为比值较高(平均为1.70)、居中(平均为1.25)和较低(平均为0.79)3类,为诱变育种选育优质专用大豆新品种提供了依据。     

Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.     

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.     

小麦Glu-3位点基因拷贝数的变异分析

【目的】基因拷贝数变异是一种常见又重要的基因结构变异,往往影响个体表型。低分子量麦谷蛋白（low-molecular-weight glutenin subunit,LMW-GS）是小麦贮藏蛋白的主要组成部分,位于Glu-3位点。小麦作为异源六倍体,其庞大且复杂的基因组结构导致难以利用传统方法检测目的基因的拷贝数,针对小麦基因组,筛选可靠稳定的内参基因和体系,探索适合复杂基因组的拷贝数变异测定技术,测定Glu-3位点LWM-GS基因拷贝数。【方法】以Acc1为内参基因,根据基因序列设计内参引物和探针,通过定性和定量PCR测定内参基因在12个普通小麦品种中的拷贝数,分析该基因拷贝数在不同品种间的稳定性;又以小麦品种篙优2018的5个稀释浓度的基因组DNA为模板,利用qRT-PCR验证Acc1内参系统的重复性和准确性;根据Glu-A3位点LMW-GS基因序列设计特异性引物及探针,利用qRT-PCR和ddPCR 2种方法检测8个小麦品种Glu-A3位点基因拷贝数,比较后选择更优的高通量基因拷贝数检测方法;再根据Glu-B3和Glu-D3位点LMW-GS基因序列设计相应的特异性引物及探针,并利用ddPCR技术检测和分析了231份小麦品种的Glu-A3、Glu-B3和Glu-D3位点上LMW-GS基因拷贝数。【结果】Acc1在12个普通小麦品种间、同一品种5个DNA稀释浓度间的拷贝数测定结果一致,技术重复间的变异系数仅为0.07%—0.77%,所构建的Acc1内参系统稳定;比较qRT-PCR和ddPCR 2种拷贝数检测方法,8个品种所测的Glu-A3位点拷贝数结果一致,分别为3、5、3、4、3、3、3和3;且ddPCR检测重复间的变异系数为0.30%—1.67%,远低于qRT-PCR的3.14%—12.72%,更加可靠;利用ddPCR对231份普通小麦品种的Glu-A3、Glu-B3和Glu-D3位点上LMW-GS基因拷贝检测后分析发现,大多数小麦品种在3个位点上的拷贝数为4,所占频率分别为51.95%、32.03%和28.57%,Glu-3位点总拷贝数变异范围为10—21,变异系数为16.12%。【结论】Acc1内参系统具有良好的稳定性和重复性,可以用作小麦Glu-3位点和其他目的基因拷贝数检测的内参;qRT-PCR和ddPCR均可用于小麦基因拷贝数的检测,但后者更稳定、可靠,且操作简单、检测通量高。     

牦牛LHB基因的分子克隆与序列分析

采用PCR法成功克隆出牦牛LHB基因的序列,在NCBI上的登录号为DQ508150。牦牛LHB基因的cds长426bp,编码141个氨基酸的产物。同源性分析,多个物种间LHB基因编码区的序列相似性在80％以上。通过牦牛与奶牛在LH的氨基酸序列上的比对发现,存在三处残基的差异,精氨酸-谷氨酰胺、甲硫氨酸-缬氨酸、苏氨酸-丙氨酸。     

家蚕G蛋白γ1亚基(BmGγ1)的克隆及其谷胱甘肽硫转移酶融合蛋白的表达与纯化

异三元G蛋白是真核细胞感知外界信号后将信号传递到胞内的重要分子,参与生物体广泛的信号转导。为了研究家蚕体内G蛋白的生理功能及其作用机制,运用生物信息学方法预测了家蚕G蛋白γ1亚基(Gγ1)的序列,设计引物验证预测序列后,克隆了家蚕Gγ1的序列,再通过酶切克隆至表达载体pET-41b(+)后,导入E.coliBL21宿主菌中,经异丙基β-D-硫代半乳糖苷(IPTG)诱导表达重组谷胱甘肽硫转移酶(glutathione s-transferase,GST)融合蛋白,并亲和层析纯化表达产物。家蚕Gγ1重组GST融合蛋白经SDS-PAGE电泳和Western blot分析,在分子质量约36 kD处出现特异性蛋白条带,重组蛋白经GST亲和层析柱纯化后,得到了高纯度的融合蛋白,说明已经成功克隆到家蚕Gγ1基因,并在E.coliBL21中高效表达。     

Metabolite profiling of the response to high-nitrogen fertilizer during grain development of bread wheat (Triticum aestivum L.)

Wheat yield and quality are dependent largely on nitrogen (N) availability. In this study, we performed the first metabolomic analysis of the response to high-N fertilizer during wheat grain development using non-targeted gas chromatography-mass spectrometry (GC–MS). Quality parameter analyses demonstrated that high-N fertilizer application led to a significant increase in grain protein content and improvement in starch and bread-making quality. Comparative metabolomic profiling of six grain developmental stages resulted in identification of 74 metabolites, including amino acids, carbohydrates, organic acids and lipids/alcohol, which are primarily involved in carbon and N metabolism. Under high-N fertilizer treatment, numerous metabolites accumulated significantly during grain development. Principal component analysis revealed two principal components as being responsible for the variances resulting from N-fertilizer treatments. Metabolite–metabolite correlation analysis demonstrated that the high-N treatment group had a greater number of positive correlations among metabolites, suggesting that high-N fertilizer treatment induced a concerted metabolic change that resulted in improved grain development. Particularly, the high-N treatment-mediated significant accumulation of metabolites involved in the TCA cycle, starch and storage protein synthesis could be responsible for the improvement of grain yield and quality. Our results provide new insight into the molecular mechanisms of wheat grain development and yield and quality.     

Allelic variation in high-molecular-weight glutenin subunit Loci of Glu-1 in Japanese common wheats

Seed storage proteins of 131 Japanese Norin wheat (Triticum aestivum) varieties were fractionated by sodium dodecyl sulfate polyacrylamide gel electrophoresis to determine allelic make-up in varieties at each of three loci that control high-molecular-weight (HMW) glutenin subunits. Three alleles were identified at the Glu-A1 locus, six at the Glu-B1 locus and five at the Glu-D1 locus. Twenty-four different, major glutenin HMW subunits were identified and each contained three to five subunits and seventeen different glutenin subunit patterns were observed for 19 subunits in the 131 Japanese Norin varieties. Fourteen alleles were identified by comparison of subunit mobility with that previously found in hexaploid wheat. Japanese Norin varieties showed a specific pattern of allelic variation in glutenin HMW subunits, different from that of Chinese and other country common wheats in allelic frequency at Glu-1 loci. This revised version was published online in July 2006 with corrections to the Cover Date.