Changes in the glutathione thiol-disulfide status in wheat grain by foliar sulphur fertilization: consequences for the rheological properties of dough

Storage proteins and glutathione in wheat play an important role in gluten network formation and can be modified by supplementation of nitrogen (N) and sulphur (S) in wheat plants. The glutathione thiol-disulfide status and its relationship to the molecular weight distribution wheat polymeric protein and dough rheological properties have been examined after different foliar S fertilizations (S derived from micronized elemental S and NS, a mixture of N urea and elemental S) applied at the post-anthesis stage. Changes in levels of reduced glutathione (GSH), glutathione disulfide (GSSG), polymeric protein-glutathione mixed disulfide (PPSSG) were analysed by reversed phase high performance liquid chromatography, during grain development using the wheat cultivars, Soissons and Trémie. During the grain desiccation phase, S supplementation (i) increased the GSSG/GSH ratio by 23–25% (ii) induced PPSSG accumulation, and (iii) decreased the formation of SDS-unextractable polymeric protein (UPP) and its molecular mass distribution. However, simultaneous N and S supplementation results in: (i) a decrease in PPSSG formation by 20–30% and (ii) an increase of UPP by 7–18% by enhancing both the branching of the aggregated proteins and their molecular weight. The mixograph parameters show that all forms of endogenous glutathione are linked to dough weakening and are negatively correlated with dough mixing tolerance, dough strength and consistency, while UPP is positively correlated with dough strength and consistency. These findings indicate that S nutrition influences dynamics of the glutathione forms in the grain and results in modification the degree of polymerization of storage protein. Thus both the changes in the form of glutathione and protein polymerization influence the rheological properties of dough.     

表达大肠杆菌K88ac-ST1-LTB融合蛋白工程菌株的免疫原性研究

已构建的能表达大肠杆菌K88ac-ST1-LTB融合蛋白的工程菌株BL21（DE3）（pXKST3LT5)及其表达产物经动物试验证实没有毒性反应。用从IPTG诱导的工程菌中提取的包涵体或经甲醛灭活的工程菌制成抗原，免疫小鼠，结果免疫小鼠至少能抵抗2MLD的大肠杆菌强毒株C83902（K88ac,ST^ ,L^ )的攻击，用提取的包涵体免疫家兔后，采集的血清能够中和天然ST1的毒性，这表明构建的工程菌株BL21（DE3）（pXKST3LT5)可以作为预防幼畜大肠杆菌性腹泻基因工程菌苗的候选株。     

小麦远缘杂交后代节燕98-2分离类型贮藏蛋白的研究

采用A-PAGE和SDS-PAGE聚丙烯酰胺凝胶电泳方法,对远缘组合分离出来的节燕98-2类型入选11个遗传性基本稳定的具有高产、多抗的小麦株系的醇溶蛋白和高分子量谷蛋白亚基进行了分析.结果表明,在A-PAGE电泳分析中,11个供试株系具有11种不同的醇溶蛋白带型.在SDS-FAGE电泳分析中,出现了7种不同的高分子量谷蛋白亚基(HMW-GS)及6种亚基组合类型,优质亚基及亚基组合所占的比例较少,品质评分偏低,其变幅为5～8分,平均为6.36分.但在所分析的材料中,出现了一个少见的特殊亚基:2 10 12.并研究了这些HMW-GS和组合频率及特点.11个株系中7个具有45 10优质亚基和2个具有2*亚基,它们可供小麦优质育种利用.研究表明,通过远缘杂交能够选育出具有高产、抗病和优质的小麦新材料.     

高分子量麦谷蛋白亚基的品质效应研究进展

介绍了小麦高分子量谷蛋白亚基(HMW—GS)的遗传，不同基因位点及位点内各等位亚基对品质的效应等研究进展，提出了通过回交转育创建几套不同高分子量谷蛋白亚基的近等基因系群，然后在同一遗传背景和不同遗传背景条件下分析高分子量谷蛋白亚基及其组合的品质效应及与遗传基础的关系。     

Molecular characterization of Ascaridia galli from Bangladesh and development of a PCR method for distinguishing A. galli from Heterakis spp.

We analyzed the nuclear ribosomal internal transcribed spacer (ITS) 1 and ITS2 sequences for Bangladesh isolates of Ascaridia galli, and we determined that the sequences were unreliable as molecular markers for distinguishing A. galli from other Ascaridia species, because the sequences showed high identity with that of A. columbae. However, the ITS1 sequences were available for designing PCR primers distinguishable between Ascaridia galli and Heterakis spp. Bangladesh isolates of A. galli constituted a monophyletic clade along with other geographical isolates in the cytochrome c oxidase subunit I (COI) phylogenetic tree, however, we could not clarify the phylogenetic relationships between A. galli and other Ascaridia spp., because their available sequences in GenBank were very few. The developed PCR method using DNA from A. galli and Heterakis spp. eggs would enable differential diagnosis of the individual infections in the future.     

拟南芥VHA-c基因在非生物胁迫响应中的作用

利用RT-PCR技术检测VHA-c基因对高盐、水和ABA胁迫的响应,结果表明所有的VHA-c基因都参与了V-ATPase的抗盐胁迫反应;在响应水胁迫的反应中VHA-c2、VHA-c4和VHA-c5基因的表达量增加;在ABA胁迫反应中VHA-c1,VHA-c2,VHA-c3和VHA-c4基因的表达水平均提高,但是VHA-c1和VHA-c3基因只在早期的ABA胁迫中增加其表达量。     

大肠杆菌k88ac菌毛蛋白亚基因的克隆、表达及亲和层析纯化

利用PCR技术,从E.coli C83902中扩增出不含信号肽序列的K88ac菌毛蛋白亚基基因片段,将其克隆到表达载体pQE-30中,构建了原核表达载体pQE30-K88ac,并转入E.coli XL1-Blue中。经IPTG诱导后,由T5启动子调控表达了氨基端带6个连续组氨酸残基的以包涵体形式存在的K88ac蛋白,在变性条件下对目的蛋白进行纯化,并获得了高纯度的融合蛋白。     

小麦低分子量谷蛋白一级结构分析

用数学统计方法和蛋白质序列分析软件对部分已测序的17种低分子量走谷蛋白亚基含氮量、氨基酸含量及一级结构序列进行比较分析。以期从氨基酸层次上揭示其结构和功能及进化上的关系．结果表明：(1)其平均含氮量为17．8％;(2)氨基酸含量基本恒定。以谷氨酰胺的含量最多。且常连接在一起;(3)存在119个保守性氨基酸残基扣两个保守性片段。一个是“QQQ-PPFSQQ”。一个是“IP-VHPSILQQLNPCKVFLQQ—C—P—AM-Q-LARSQM-QS-CHVMQQQCCQQL—QIPQQSRYEAI-AI-YSIILQEQQ”(“-”表示各序列中不同的氨基酸);(4)X84960、X84961、Y14104;X13306、U86025、U86027、U86029两组亚基序列中各具有一定程度的同源性。     

Metabarcoding of feces and intestinal contents to determine carnivorous diets in red-crowned cranes in eastern Hokkaido,Japan

The red-crowned crane Grus japonensis in Hokkaido, Japan forms a closed population as a residence that is independent of the mainland population. Based on observations of a limited number of individuals as well as cranes in captivity, red-crowned cranes are omnivores and eat fish, worms, insects and plants in their own territories except in winter, when they are fed with dent corn that is supplied in eastern Hokkaido. DNA metabarcoding based on high throughput sequencing was carried out using universal primer sets for cytochrome oxidase subunit I gene. Feces from 27 chicks collected in June and July in the period from 2016 to 2018 and intestinal contents from 33 adult and subadult cranes that were found dead almost throughout year in 2006–2013 in the field in eastern Hokkaido were used. Although compositions varied considerably in the cranes, both insects and fish were found in adults and subadults to the same extents, while insects were predominant in chicks. Both insects and fish were detected in all seasons for adults and subadults. Horse flies, scarab beetles and weevils accounted for the most of the insects regardless of the life stage. Dace, stickleback, flatfish and sculpin were the major fish species in adults, while chicks ate almost only stickleback. The results provide the first comprehensive data on carnivorous diets in wild red-crowned cranes in eastern Hokkaido as basis for conservation of red-crowned cranes, for which the life style and area continue to change.