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)。     

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.     

猪链球菌噬菌体SMP末端酶大亚基单位的活性研究

末端酶(terminase)是dsDNA病毒包装过程中的必要功能蛋白,其大亚基单位在包装过程中往往行使全酶的作用,一般具有ATP酶活性和限制性内切酶活性。噬菌体SMP是猪链球菌2型的烈性噬菌体,本研究以SMP基因组为模板,扩增SMP末端酶大亚基单位基因(TlsSMP),构建pEASY-E1-Tls重组质粒,在大肠杆菌中诱导表达,并对TlsSMP融合蛋白进行活性检测。结果显示,末端酶大亚基单位在大肠杆菌中呈可溶性表达,具有ATP酶生物学活性,在pH7、23℃和Mg2+存在的条件下,其活性最强,其他金属离子如Ca2+、Mn2+和Co2+都对其有不同程度的抑制作用。本研究为进一步研究猪链球菌噬菌体的包装机制奠定基础。     

刺五加ATP合酶β亚基基因的克隆与序列分析

[目的]克隆刺五加叶绿体的ATP合酶β亚基cDNA并对其进行生物信息学分析。[方法]根据已知物种叶绿体ATP合酶β亚基基因的序列,设计1对同源引物,通过RT-PCR扩增得到刺五加ATP合酶β亚基cDNA,并对其进行序列比对及结构预测分析。[结果]RT-PCR扩增获得了长1099bp的刺五加ATP合酶β亚基cDNA,该基因编码366个氨基酸。序列比对及结构预测分析表明,刺五加ATP合酶β亚基基因编码的氨基酸与水稻的同源性最高,达96.41%。其二级结构中含有171个α螺旋（alpha helix）,占46.72%;53个延伸链（extended strand）,占14.48%;27个β折叠（beta turn）,占7.38%;115个无规则蜷曲（random coil）,占31.42%。第262～271位氨基酸为ATP合酶β亚基的标志性位点。整个多肽链无明显的疏水区域,初步认定为亲水性蛋白。[结论]该试验克隆得到的ATP合酶β亚基基因为叶绿体ATP合酶β亚基基因,为研究刺五加能量代谢对植物次生代谢的影响及了解植物ATP合酶的结构与功能提供了必要的信息。     

江苏省小麦品种的谷蛋白亚基组成分析

为了解江苏省小麦品种的谷蛋白亚基组成现状,为品质育种提供理论指导,选用77份来自江苏省淮南和淮北主要小麦育种单位的育成品种或高代品系,利用聚丙烯酰胺凝胶电泳法(SDS-PAGE)对Glu-1和Glu-3位点进行了鉴定。结果表明,参试品种(系)Glu-1和Glu-3位点的高分子量麦谷蛋白亚基(HMW-GS)和低分子量麦谷蛋白亚基(LMW-GS)变异丰富。共检测到12个HMW-GS等位变异,其中Glu-A1位点有3个,Glu-B1位点有5个,Glu-D1位点有4个。LMW-GS的Glu-A3和Glu-B3位点则分别检测到5个和6个等位变异。淮北小麦的HMW-GS多态性高于淮南小麦。Glu-1和Glu-3位点等位变异的分布频率差异较大,且淮北和淮南表现不同,优质亚基比例较低。Glu-1位点主要亚基类型为0(46.8%)、1(49.4%)、7+8(63.6%)和2+12(63.6%),Glu-A3位点主要亚基类型为a(23.4%)、c(57.1%)和d(15.6%),Glu-B3位点主要亚基类型为b(22.1%)、f(49.4%)和j(1B/1R易位)(16.9%)。淮北小麦的高分子量谷蛋白优质亚基及亚基组合较少,Glu-B3j分布频率高(54.5%),亚基质量有待提高;淮南小麦的优质亚基及组成亦较少,需根据中筋和弱筋小麦育种目标并结合蛋白质含量和亚基选择进行改良。     

河北省小麦品种高分子量谷蛋白亚基组成分析

高分子量麦谷蛋白(HMW-GS)亚基组成是决定小麦加工品质的重要蛋白组分,对小麦品质具有重要影响。本研究利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)方法分析了148份河北省小麦品种的高分子量麦谷蛋白亚基(HMW-GS)组成及其历史变化。结果表明:在148份小麦品种中共检测到16种HMW-GS亚基和43种亚基组合。在Glu-A1位点,有3种亚基类型,以null和1亚基为主;Glu-B1位点有7种亚基类型,以"7+9"和"7+8"亚基为主;Glu-D1位点有6种亚基类型,以"2+12"亚基类型为主。在43种亚基组合中,以"null、7+9、2+12"组合为主,其他亚基组合尤其是与品质有关的亚基组合相对较少。从河北省小麦品种HMW-GS的历史演变来看,优质蛋白亚基及其组合虽然近年来有所提高,但仍需进一步加强。     

陕西省部分小麦栽培品种高分子量麦谷蛋白亚基组成及其与品质关系

本研究以36个陕西省主要栽培品种为材料,利用SDS-PAGE方法分析了高分子量麦谷蛋白亚基组成,并对其SDS沉淀值进行了测定,分析了高分子量麦谷蛋白亚基组成与代表面包烘烤品质的SDS沉淀值间的关系.结果表明:Glu-1位点存在着广泛的等位基因变异,在所分析的36个品种中,Glu-A1位点亚基1出现频率最高,为63.9%;Glu-B1位点等位基因变异最丰富,其中亚基7 8和7 9出现频率最高,其次为亚基14 15;Glu-D1位点亚基组成以2 12为主;对各位点品质评分与SDS沉淀值的简单相关分析结果表明各位点品质评分与SDS沉淀值均存在着显著或极显著相关关系,各位点影响大小依次为Glu-D1＞Glu-A1＞Glu-B1;Glu-1品质评分与SDS沉淀值极显著正相关.     

重庆小麦品种(系)的高分子量麦谷蛋白亚基(HMW-GS)组成分析

为了解重庆小麦的品质遗传基础,采用SDS-PAGE技术对重庆小麦推广品种和主要育种亲本共35份材料的HMw-GS组成和变异进行了分析.结果表明:参试材料Glu-1位点具有较为丰富的遗传变异,并检测到14种亚基类型和19种亚基组合类型.Glu-A1位点检测到2种亚基(1,null),其中null占82.86%;Glu-B1位点检测到9种亚基(7+8,7+9,7,22,6+8,14+15,17+18,20,8),其中以7+8,7+9出现的频率最高;Glu-D1位点检测到3种亚基(2+12,5+10,4+12),其中5+10占57.14%;参试材料中检测到与烘烤品质相关的优质亚基.参试材料的品质评分在4～10分之间,平均分为6.49分.在重庆市的7个小麦推广品种中,检测到6种亚基组合类型,品质评分平均为6.86分.     

Combination of serum phosphorylated neurofilament heavy subunit and hyperintensity of intramedullary T2W on magnetic resonance imaging provides better prognostic value of canine thoracolumbar intervertebral disc herniation

The aim of this study was to evaluate the prognostic value of concurrent measurement of serum phosphorylated neurofilament heavy subunit (pNF-H) concentration and intramedullary T2W hyperintensity in paraplegic to paraplegic dogs. Our hypothesis was that concurrent measurement of these would provide a more accurate prediction of functional outcome in dogs with thoracolumbar intervertebral disc herniation (IVDH). A prospective case-control clinical study was designed using 94 dogs with acute onset of thoracolumbar IVDH. The association of serum pNF-H concentration, T2W hyperintensity on sagittal MRI (T2H/L2), deep pain perception and surgical outcome were evaluated with logistic regression analysis after three months for all 94 surgically treated dogs. Sensitivity to predict non-ambulatory outcome was compared among pNF-H and T2H/L2 and combination of both. Logistic regression analysis indicated that serum pNF-H concentration and T2H/L2 were significantly correlated with surgical outcome (P<0.05); however, deep pain perception was not (P=0.41). The results of logistic regression analysis indicated that the odds ratios of unsuccessful long-term outcome were 2.6 for serum pNF-H concentration, 1.9 for T2H/L2 and 2.3 for deep pain sensation. The sensitivity and specificity to predict non-ambulatory outcome for using serum parameter pNF-H>2.6 ng/ml, using T2H/L2 value of>0.84 and using both serum pNF-H and T2H/L2, were 95% and 75.7%, 65% and 86.5%, and 90.0% and 97.5%, respectively. Therefore, combined measurements of serum pNF-H and T2H/L2 might be useful for predicting long-term outcome in dogs with thoracolumbar IVDH.     

Downregulation of the expression of inhibin α subunit and betaglycan in porcine cystic follicles

Inhibins, as members of the transforming growth factor beta (TGF-β) superfamily, downregulate the synthesis and secretion of follicle-stimulating hormone (FSH) in an endocrine manner. The role of inhibin/betaglycan in the ovary regulation recently gained attention. To date, no data exist on the function of inhibin α subunit and betaglycan in cystic follicles. In this study, the expressions of inhibin α subunit and betaglycan in cystic follicles were investigated using immunohistochemistry, real-time PCR and Western blot analysis. Both inhibin α subunit and betaglycan immunoreactivities were mainly localized in the granulosa cells of follicles. Expression of inhibin α subunit and betaglycan was inferior in cystic follicles compared with that in normal large follicles. However, the result of enzyme-linked immunosorbent assay showed no significant difference in the decreasing in concentration of inhibin α subunit in cystic follicular fluid compared with the control (P>0.05). In this study, we explored the effects of FSH on betaglycan expression in granulosa cells in vitro. As expected, a significant increase in the expressions of betaglycan mRNA and protein in granulosa cells was observed in response to exogenous FSH (30 ng/ml) (P<0.05) compared with the control. Consequently, this study provides evidence that the expressions of inhibin α subunit and betaglycan are inferior in cystic follicles, and this may be caused by the decrease in FSH in the presence of a cystic follicle.