施氮水平对7S亚基缺失大豆根系形态和结瘤固氮的影响

为有效推广功能型大豆7S亚基缺失品种,以7S亚基缺失大豆品系东富2号为研究对象,设置4种施氮水平(纯N),N0(0 mg·kg~(-1))、N1(25 mg·kg~(-1))、N2(50 mg·kg~(-1))、N3(75 mg·kg~(-1)),采用桶栽法研究大豆根系形态和结瘤固氮对不同施氮水平的响应。结果表明:N1(25 mg·kg~(-1))水平下根系干重加大,根冠比增大,根瘤固氮潜力高,单株产量较高。N2(50 mg·kg~(-1))水平下根长、根表面积、根体积在生育后期增长较快,根系干重较大,根冠比低,固氮酶活性最高,单株籽粒产量最高。N3(75 mg·kg~(-1))水平下植株干重较大,无效生长较多,根瘤数少,固氮潜力和根冠比低,单株产量不高。综合籽粒产量和根系特性指标,功能型大豆7S亚基缺失品系东富2号的适宜施肥量为25~50 mg·kg~(-1)。     

Kernel vitreousness and protein content: Relationship,interaction and synergistic effects on durum wheat quality

Four sets of durum samples were used in this study to further understand the interrelationships among hard vitreous kernels (HVK), protein content, and pigment concentration, with a focus on the interaction and synergistic effects of protein content and vitreousness on durum quality. HVK level increases with higher protein content in the range of 9.5–12.5%, but this relationship is less evident in durum samples with high protein content (12.5–14.5%). Both protein content and kernel vitreousness can significantly affect durum milling quality. White starchy kernels (WSK) in low protein durum have a very detrimental impact on milling and pasta processing quality, but high protein content can mitigate the adverse impact of WSK on durum quality. Although protein content plays a dominant role, higher HVK might contribute positively to pasta firmness. There was no significant difference in yellow pigment content between HVK and WSK. However, pigment loss from semolina to dough was higher for WSK than HVK. Despite the difference in protein content, HVK and WSK have little difference in gluten strength. The monomeric protein was preferentially accumulated in HVK. The glutenin proteins of HVK and WSK were similar in the ratios of 1Bx/1By and HMW/LMW-GS.     

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。     

Survey and new insights in the application of PCR‐based molecular markers for identification of HMW‐GS at the Glu‐B1 locus in durum and bread wheat

Wheat, among all cereal grains, possesses unique characteristics conferred by gluten; in particular, high molecular weight glutenin subunits (HMW‐GS) are of considerable interest as they strictly relate to bread‐making quality and contribute to strengthening and stabilizing dough. Thus, the identification of allelic composition, in particular at the Glu‐B1 locus, is very important to wheat quality improvement. Several PCR‐based molecular markers to tag‐specific HMW glutenin genes encoding Bx and By subunits have been developed in recent years. This study provides a survey of the molecular markers developed for the HMW‐GS at the Glu‐B1 locus. In addition, a selection of molecular markers was tested on 31 durum and bread wheat cultivars containing the By8, By16, By9, Bx17, Bx6, Bx14 and Bx17 Glu‐B1 alleles, and a new assignation was defined for the ZSBy9_aF1/R3 molecular marker that was specific for the By20 allele. We believe the results constitute a practical guide for results that might be achieved by these molecular markers on populations and cultivars with high variability at the Glu‐B1 locus.     

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.     

猪传染性胸膜肺炎重组亚单位疫苗诱导小鼠免疫保护效果的观察

重组表达猪传染性胸膜肺炎放线杆菌6种主要毒力因子基因:apxⅠ、apxⅡ、apxⅢ、apxⅣ、apfa和omp,以重组蛋白rApxⅠ、rApxⅡ、rApxⅢ和rOMP组合免疫小鼠作为试验I组,重组蛋白rApxⅠ、rApxⅡ、rApxⅢr、OMPr、ApxⅣ和rApfa组合免疫小鼠作为试验Ⅱ组,PBS为对照组,分3次免疫小鼠,采用背部皮下多点注射,每次间隔2周,免疫剂量为0.2 mL/只,3免后1周分别以APP1型菌Shope 4074株(5×109cfu)和APP2型菌S1536株(5×1010cfu)进行攻毒试验。通过小鼠保护率与抗体效价的相关性研究、肺部病理变化及肺脏细菌的分布情况等指标进行综合评价。结果显示,试验Ⅰ组4种重组蛋白特异性抗体水平显著高于其他两组(P<0.05),对APP1型菌攻毒的保护率(9/10)明显高于试验Ⅱ组(5/10)和对照组(0/8),小鼠的免疫保护率与抗体效价之间存在显著正相关;且该组对APP2型菌攻毒的保护作用(无肺脏损伤)也明显优于其它两组(典型肺部损伤)。间接免疫荧光试验表明试验Ⅰ组对肺脏细菌的清除效果也明显优于其他两组。本试验揭示试验Ⅰ组对不同血清型APP攻击能够提供很好的交叉保护作用,从而为猪传染性胸膜肺炎新型疫苗的研制提供参考。