应用原位杂交及RAPD技术标记抗黄矮病小麦—中间偃麦草染色体异附加系

以生物素标记中间偃麦草基因组ＤＮＡ为探针,与抗黄矮病小麦－中间偃麦草染色体异附加系Ｚ６进行原位杂交,鉴定出附加的１对中间偃麦草染色体。对异附加系Ｚ６和Ｌ１及它们的小麦亲本进行了ＲＡＰＤ分析,从１２０个随机引物中,筛选出２个引物可以扩增出附加染色体的特异ＤＮＡ片段,可作为鉴定导入小麦的中间偃麦草染色质的分子标记。     

用分子标记定位源于中间偃麦草的小麦抗黄矮病基因

利用生物素标记的中间偃麦草基因组总DNA作为探针 ,对以L1为抗源的抗黄矮病小麦新品系H960 642的有丝分裂中期染色体进行原位杂交 ,结果表明 :H960 642是纯合的小麦 中间偃麦草易位系 ,携有抗黄矮病基因的中间偃麦草染色体片段易位到小麦染色体端部 .采用小麦第 7部分同源群上的 8个RFLP探针进行Southern分析 ,结果表明 :H960 642的小麦 7D染色体长臂末端片段被中间偃麦草染色体 7X长臂末端片段所取代 ,即该染色体为T7DS·7DL -7XL ,易位断点位于Xpsr680和Xpsr965之间 ,距着丝点的遗传距离约 90～ 99cM .筛选出了与 7X上抗黄矮病基因紧密连锁的RFLP标记psr680和psr687.将该抗黄矮病基因定位于 7XL端部、RFLP遗传图Xpsr680与Xpsr687位点附近 .Ep 1同工酶分析结果佐证了RFLP分析结果 .     

中间偃麦草染色体2Ai-2特异PCR新标记的建立和St基因组特异序列的克隆

中间偃麦草麦、小麦和小麦－中间偃麦草2Ai－2附加系Z1、Z2、X6,代换系ZD28等进行RAPD分析,从320个RAPD引物中,鉴定出2Ai－2染色体特异的2个RAPD标记OPO05650和OPMO414000。利用这2个特异OPO05和OPM04,PCR扩增普通小麦CS（ABD）及其近缘植物中间偃麦草（E1E2St）、拟鹅冠草（St）,长穗偃麦草（E）、簇毛麦（V）、黑麦（R）、大麦（H）粗山羊草（D）等基因组DNA。结果表明,OPO05650和OPO41400均是2Ai－2染色体上St基因组区域的特异标记。将上棕2个特异片段分离回收、克隆、测序,根据测序结果重新设计、合成特异引物,成功地转换RAPD标记为SCAR（sequence characterizked amplifed region）标记SC－05和SC－M4。利用SCAR标记对不同材料进行分析的结果表明,凡含有2Ai－2染色体的抗黄矮病材料及拟鹅冠草均产生一条扩增带,不含2Ai－2染色体的材料,包括小麦、长穗麦草、簇毛麦、黑麦、在麦、粗山羊草以有含有其他他中间偃麦草染色休的附加系,均没有扩增产物,说明上棕2个SCAR标记是中间偃麦草2Ai－2染色体的特异性PCR标记,且是2Ai－2染色体上St基因组区域的特异性标记。克隆与鉴定中间偃麦草的2个SCAR扩增片段TiSCO5和TiSCM4。结果表明,克隆的中间偃麦草TiSCO5和TiSCM4特异片段,分别是St基因组特异性的寡拷贝序列有多拷贝重复序列,为St基因组遗传研究的新探针。     

用克隆池PCR法筛选小麦-簇毛麦易位系6VS/6AL基因组TAC文库获得抗病基因候选克隆

用根据抗病基因保守区设计的一对简并性引物,从小麦－簇毛麦易位系6VS／6AL cDNA中PCR扩增获得一个具有抗病基因核苷酸结合位点（Nucleotide binding site,NBS）结构特点的DNA片段克隆N7。从小麦－簇毛麦易位系6VS／6AL基因组TAC（Transformation-competent artificial chromosome,TAC）文库的22块96孔板提取所有2112个克隆池（每个池含约1000个克隆）的质粒,再根据N7的核苷酸序列设计一对特异引物,用克隆池PCR（pooled PCR)法经分级筛选从文库中获得一个阳性克隆。以N7为探针,通过Southern杂交证实了该TAC克隆为真正含有抗病候选基因的克隆。研究结果表明克隆池PCR法对克隆数目巨大的基因组文库的筛选很有效。     

用克隆池PCR法筛选小麦 簇毛麦易位系6VS/6AL基因组TAC文库获得抗病基因候选克隆

用根据抗病基因保守区设计的一对简并性引物,从小麦簇毛麦易位系6VS/6AL cDNA中PCR扩增获得一个具有抗病基因核苷酸结合位点（Nucleotide binding site, NBS）结构特点的DNA片段克隆N7。从小麦簇毛麦易位系6VS/6AL基因组TAC（Transformationcompetent artificial chromosome, TAC）文库的22块96孔板提取所有2112个克隆池（每个池含约1000个克隆）的质粒,再根据N7的核苷酸序列设计一对特异引物,用克隆池PCR（pooled PCR）法经分级筛选从文库中获得一个阳性克隆。以N7为探针,通过Southern 杂交证实了该TAC克隆为真正含有抗病候选基因的克隆。研究结果表明克隆池PCR法对克隆数目巨大的基因组文库的筛选很有效。     

抗黄矮病普通小麦偃麦草异附加系、异代换系的选育和鉴定

以偃麦草 (Ag .pulcherrimum)为抗源 ,以圆锥小麦 (T .turgidum)×偃麦草的双二倍体———CPI113 5 0 0 ( 2n =70 )与普通小麦杂交、回交、自交得到的衍生系为基础材料 ,利用黄矮病抗性追踪、形态学标记、细胞遗传学分析 ,筛选到 2个抗黄矮病新种质 96S16 11,96W 14 9.通过测交分析、原位杂交和同工酶电泳分析等技术 ,对以上材料进行鉴定 ,结果表明 :96S16 11为普通小麦 偃麦草二体异附加系 ,96W 14 9为普通小麦 偃麦草 1D( 1Ap)二体异代换系 .     

应用原位杂交及RAPD技术标记抗黄矮病小麦一中间偃麦草染色体异附加系

以生物素（Ｂｉｏｔｉｎ－１６－ｄＵＴＰ）标记中间偃麦草基因组 ＤＮＡ为探针,与抗黄矮病小麦－中间偃麦草染色体异附加系Ｚ６进行原位杂交,鉴定出附加的１对中间偃麦草染色体。对异附加系 Ｚ６和 Ｌ１及它们的小麦亲本进行了 ＲＡＰＤ分析,从 １２０个随机引物中,筛选出 ２个引物可以扩增出附加染色体的特异ＤＮＡ片段,可作为鉴定寻人小麦的中间偃麦草染色质的分子标记。     

普通小麦-中间偃麦草TAI-27中附加染色体的显微切割及特异性探针的筛选

试验以长穗偃麦草基因组DNA为探针 ,与普通小麦 中间偃麦草TAI 2 7进行染色体原位杂交 ,表明有 4条与长穗偃麦草同源的染色体 ;以P .stipifolia (St)基因组DNA为探针 ,有 4条与St同源的染色体 .这说明TAI 2 7中有 4条St染色体 .TAI 2 7是异代换 附加系 .对TAI 2 7中附加的中间偃麦草染色体进行显微切割 ,并建立其微克隆库 ,从中筛选获得了中间偃麦草的特异性探针 ,同源性分析表明该序列为一新序列 .这为进一步筛选抗病、抗逆和优质基因打下基础 .     

利用花药培养快速创制小麦条锈病和黄矮病抗性新种质

利用中间偃麦草与普通小麦杂交育成的部分双二倍体－中５为外源抗性基因供体,通过花药培养途径,在它与几个冬小麦品种的５个杂交组合中,诱导出试管花粉绿苗１４４株,移栽加倍后,形成了４９个加倍单倍体株系。通过小麦条锈病和黄矮病抗性鉴定获得了几个高抗黄矮病或者对条锈病近免疫的新材料。其中ＤＨ７２８对条锈病菌近免疫,２ｎ＝４４,减数分裂构型为０．７０１Ｖ＋０．０３５ＩＩＩ＋２１．５７９ＩＩ＋０．４５６Ｉ,为双     

小麦-簇毛麦6VS/6AL易位系可转化人工染色体(TAC)文库的构建

可转化人工染色体(Transformation\|competent Artificial Chromosome,TAC)是具有克隆和转移大片段基因能力的新型载体,是植物基因克隆和转化的有效工具。为了克隆小麦抗白粉病基因和其它基因,本研究用TAC载体pYLTAC17构建了带有抗白粉病基因Pm21的小麦簇毛麦6VS/6AL易位系的基因组DNA文库。该文库包含210万个克隆,平均插入片段35kb,库容相当于普通小麦基因组的49倍。本文库以约1000个克隆组成1个混合池的方式保存在22块96孔平板,可用PCR方法进行文库的筛选。     

Tuning of electronic properties of one-dimensional cyano-bridged Cu-Ni, Cu-Pd, and Cu-Pt bimetallic assemblies by stereochemistry of ligands

Preparations, XPS and electronic spectroscopy, and magnetism of seven new one-dimensional cyano-bridged coordination polymers, chiral [Cu(RR-chxn)₂][Pd(CN)₄] · 2H₂O (1), [Cu(trans-chxn)₂][M(CN)₄] · 2H₂O (2, 4, and 6 for M = Pd, Ni, and Pt), and [Cu(cis-chxn)₂][M(CN)₄] · 2H₂O (3, 5, and 7 for M = Pd, Ni, and Pt) (RR-chxn = cyclohexane-(1R,2R)-diamine, trans-chxn = racemic trans-cyclohexane-(1,2)-diamine, and cis-chxn = racemic cis-cyclohexane-(1,2)-diamine) have been reported in view of tuning of their electronic properties by stereochemistry of chxn ligands and metal-substitution. Comparison of Cu 2p_1/2 and 2p_3/2 peaks of XPS and broad d-d bands around 18 000 cm⁻¹ of electronic spectra are described systematically for 1-7. Variable-temperature magnetic measurement shows that complexes 1-7 indicate weak antiferromagnetic interactions via cyano-bridges. Because of semi-coordination coupled with pseudo Jahn-Teller elongation and electrostatic interaction for 1, the axial Cu-N coordination bond distances of 2.330(7) and 3.092(8) Å are considerably longer than those of equatorial ones in the range from 2.016(6) to 2.030(6) Å. The former bond distances of 1 are intermediate values among the related Ni (2.324(6) and 3.120(8) Å) and Pt (2.34(1) and 3.09(1) Å) complexes.     

Structure and function of S-adenosylhomocysteine hydrolase

In mammals, S-adenosylhomocysteine hydrolase (AdoHcyase) is the only known enzyme to catalyze the breakdown of S-adenosylhomocysteine (AdoHcy) to homocysteine and adenosine. AdoHcy is the product of all adenosylmethionine (AdoMet)-dependent biological transmethylations. These reactions have a wide range of products, and are common in all facets of biometabolism. As a product inhibitor, elevated levels of AdoHcy suppress AdoMet-dependent transmethylations. Thus, AdoHcyase is a regulator of biological transmethylation in general. The three-dimensional structure of AdoHcyase complexed with reduced nicotinamide adenine dinucleotide phosphate (NADH) and the inhibitor (1′R, 2′S, 3′R)-9-(2′,3′-dihyroxycyclopenten-1-yl)adenine (DHCeA) was solved by a combination of the crystallographic direct methods program, SnB, to determine the selenium atom substructure and by treating the multiwavelength anomalous diffraction data as a special case of multiple isomorphous replacement. The enzyme architecture resembles that observed for NAD-dependent dehydrogenases, with the catalytic domain and the cofactor binding domain each containing a modified Rossmann fold. The two domains form a deep active site cleft containing the cofactor and bound inhibitor molecule. A comparison of the inhibitor complex of the human enzyme and the structure of the rat enzyme, solved without inhibitor, suggests that a 17° rigid body movement of the catalytic domain occurs upon inhibitor/substrate binding.     

Influence of Fluoride on Rat Kidney Antioxidant System: Effects of Methionine and Vitamin E

The aim of the study has been to determine and compare the influence upon the kidney antioxidative system, exercised by administration of vitamin E, and vitamin E in combination with methionine, under conditions of oxidative stress induced by sodium fluoride. The experiment was carried out on Wistar FL rats (adult males) that, for 35 days, were administered water, NaF, NaF with vitamin E, or vitamin E with methionine (doses: 10 mg NaF/kg of body mass/24 h, 3 mg vitamin E per 10 μl per rat for 24 h, 2 mg methionine per rat for 24 h). The influence of administered sodium fluoride and antioxidants upon the antioxidative system in kidney was examined by analyzing the concentration of malondialdehyde (MDA) and the activity of the most important antioxidative enzymes (SOD, total and both its isoenzymes, GPX, GST, GR, and CAT). The studies carried out confirmed the disadvantageous effect of the administered dose of NaF upon the antixodiative system in rats (increase in the concentration MDA, decrease activity of all antioxidative enzymes). The administration of vitamin E increased the activity of studied enzymes with the exception of glutathione reductase GR; it also reduced the procesess of lipid peroxidation. It has been found that combined doses of vitamin E and methionine were most effective in inhibiting lipid peroxidation processes. The results confirmed the antioxidative properties of methionine.     

Reorientation of Microfibrils and Microtubules at the Outer Epidermal Wall of Maize Coleoptiles During Auxin-Mediated Growth

Auxin-mediated elongation growth of isolated subapical coleoptile segments of maize (Zea mays L.) is controlled by the extensibility of the outer cell wall of the outer epidermis (Kutschera et al., 1987). Here we investigate the hypothesis that auxin controls the extensibility of this wall by changing the orientation of newly deposited microfibrils through a corresponding change in the orientation of cortical microtubules. On the basis of electron micrographs it is shown that cessation of growth after removal of the endogenous source of auxin is correlated with a relative increase of longitudinally orientated microfibrils and microtubules at the inner wall surface. Conversely, reinduction of growth by exogenous auxin is correlated with a relative increase of transversely orientated microfibrils and microtubules at the inner wall surface. These changes can be detected 30–60 min after the removal and addition of auxin, respectively. The functional significance of directional changes of newly desposited wall microfibrils for the control of elongation growth is discussed.     

Land,livestock, and livelihoods: Changing dynamics of gender,caste, and ethnicity in a Nepalese Village

Over the past 10 years, Ghusel VDC, Lalitpur District has moved from primarily subsistence agriculture into the wider cash economy aided by the Small Farmers' Development Program (SFDP), which provides credit to farmers mainly for the purchase of buffalo for milk production, and by the National Dairy Corporation, which supports local dairy cooperatives. Analysis reveals that buffalo-keeping and milk sales are increasing the well-being of many households, while at the same time creating new inequalities in gender roles and responsibilities, greater inequities between Brahmin and Tamang residents in Ghusel, and placing pressures on the ecosystem for increased supplies of fodder and fuelwood. Evidence suggests that there is critical, need for attention to the social, and particularly gender-based, implications of maintaining livestock for milk sales and to the ecological underpinnings of this livelihood system.     

Chiral Separation of G‐type Chemical Warfare Nerve Agents via Analytical Supercritical Fluid Chromatography

Chemical warfare nerve agents (CWNAs) are extremely toxic organophosphorus compounds that contain a chiral phosphorus center. Undirected synthesis of G‐type CWNAs produces stereoisomers of tabun, sarin, soman, and cyclosarin (GA, GB, GD, and GF, respectively). Analytical‐scale methods were developed using a supercritical fluid chromatography (SFC) system in tandem with a mass spectrometer for the separation, quantitation, and isolation of individual stereoisomers of GA, GB, GD, and GF. Screening various chiral stationary phases (CSPs) for the capacity to provide full baseline separation of the CWNAs revealed that a Regis WhelkO1 (SS) column was capable of separating the enantiomers of GA, GB, and GF, with elution of the P(+) enantiomer preceding elution of the corresponding P(–) enantiomer; two WhelkO1 (SS) columns had to be connected in series to achieve complete baseline resolution. The four diastereomers of GD were also resolved using two tandem WhelkO1 (SS) columns, with complete baseline separation of the two P(+) epimers. A single WhelkO1 (RR) column with inverse stereochemistry resulted in baseline separation of the GD P(–) epimers. The analytical methods described can be scaled to allow isolation of individual stereoisomers to assist in screening and development of countermeasures to organophosphorus nerve agents. Chirality 26:817–824, 2014. © 2014 The Authors. Chirality published by John Wiley Periodicals, Inc.     

In vitro studies of the rabbit immune system. IV. Differential mitogen responses of isolated T and B cells.

The mitogenic responses of separated rabbit lymphocyte populations functionally analogous to mouse T and B cells have been tested in vitro. Purified T cells were prepared by passage over nylon wool (NW) and purified B cells prepared by treatment with antithymocyte serum and complement (ATS + C). ATS + C kills 70% of peripheral blood lymphocytes (PBL's) and 50% of the spleen cells while passage over NW yields 40% of the applied PBL's and 5–23% of the applied spleen cells. NW-purified T cells from the spleen or PBL's respond fully to concanavalin A (Con A) but have a reduced response to phytohemaglutinin (PHA) and little or no response to goat anti-rabbit immunoglobulin (anti-Ig). PBL's that survive ATS + C (B cells) are stimulated by anti-Ig but not by Con A or PHA. B cells purified from spleen do not respond to Con A or PHA but will respond to anti-Ig under appropriate conditions. A full spleen B-cell response to anti-Ig required removal of Ig produced by the cultures that blocked anti-Ig stimulation. It is concluded that, for rabbit lymphocytes, Con A and PHA are primarily T-cell mitogens and that anti-Ig is primarily a B-cell mitogen. However, the mitogen response of unfractionated PBL or spleen cell populations indicates an overlap in reactivity. This could be due to cells sharing T and B properties, alteration of cell populations by the fractionation procedures used, or recruitment of one population in the presence of a mitogenic response of the other population.     

邻苯二甲酸二丁酯对翡翠贻贝抗氧化酶及脂质过氧化水平的影响

实验室条件下,研究了不同浓度邻苯二甲酸二丁酯(DBP)长期胁迫(15 d)对翡翠贻贝内脏团和外套膜抗氧化酶(超氧化物歧化酶SOD、过氧化氢酶CAT)及脂质过氧化(LPO)水平(以MDA含量表示)的影响,以及受胁迫翡翠贻贝在清洁海水中恢复阶段上述生化指标的变化特征.结果表明:胁迫阶段,0.5和2.5 mg.L-1DBP下翡翠贻贝内脏团SOD活性表现为先抑制后逐渐恢复,12.5和62.5 mg.L-1下则持续受到显著抑制;不同浓度组CAT活性均明显被抑制.LPO水平明显升高.外套膜中,2.5 mg.L-1下SOD活性受到持续诱导,其他浓度组则先被抑制,后随曝露时间延长逐渐被诱导;各浓度组CAT的变化波动较大,没有明显规律;而LPO水平明显升高.净化恢复阶段,12.5和62.5 mg.L-1DBP胁迫下的内脏团SOD和CAT活性恢复较慢,其LPO水平随时间延长逐渐恢复至对照组水平;外套膜中SOD活性呈持续升高趋势,CAT活性和LPO水平则随时间延长恢复到对照组水平.     

多马胺能药物对鲇鱼促性腺激素（GtH）分泌活动的影响

以珠江流域鲇鱼（silurus asotus)为实验材料,研究了多巴胺（DA）能药物（DA及其D－2型受体拮抗物 ,DOM）对鲇鱼促性腺激素（GtH)释放的影响,结果表明,在性腺发育的各个时期,单独注射DOM（5ug/g)均不能显著提高鲇鱼血液基础GtH水平,当DOM与LHRH－A联合注射时能显著增强LHRH－A刺激GtH释放的作用;DA只能抑制GnRH诱导的GtH释放,对基础GtH释放无抑制作用,这种生殖内分泌调节方式与鲇形目的革胡子鲇（Clarias gariepinus)和大鳍Hu（Mystus macropterus)相似,而与鲤形目的鲁科（Cyrpindiae)鱼类不同。     

X-ray induction of dominant lethals in late spermatids and mature spermatozoa of Drosophila melanogaster: the role of oxygenation

The role of oxygenation in determining the sensitivity to the induction of dominant lethals was examined in late spermatids and mature spermatozoa of Drosophila melanogaster. 0–2-h-old or 7-day-old Oregon-K males were irradiated with different X-ray exposures in nitrogen, air or in oxygen and the frequencies of dominant lethals induced in these stages were studied. The results obtained confirm and extend Sobels' earlier observations and the interpretation derived therefrom namely, that under normal conditions in air, mature spermatozoa are characterised by a higher degree of oxygenation than late spermatids and this difference is sufficient to explain the differential response of these stages. Similar Oxygen-Enhancement-Ratios(OERs) (of about 2) were obtained for both the cell stages. The present data also revealed that late spermatids are significantly less sensitive than mature spermatozoa to the X-ray-induction of dominant lethals in a nitrogen atmosphere. A plausible mechanism is suggested to explain this observation.