来自易变山羊草抗禾谷孢囊线虫基因的克隆与序列分析

禾谷孢囊线虫（Heterodera avenae）是严重危害禾谷类作物的病原线虫之一,它广泛分布于澳大利亚、欧洲、北美、印度和中国等世界主要小麦产区,使作物严重减产,造成巨大的经济损失。目前最有效的防治措施之一是将外源抗性基因导入栽培小麦(Triticum aestivum L.)，培育抗禾谷孢囊线虫的新品种。但迄今为止抗禾谷孢囊线虫基因克隆研究的相关报道却很少。 本实验根据此前从抗禾谷孢囊线虫材料E-10扩增得到的与来自节节麦（Aegilops tauschii）的抗禾谷孢囊线虫基因Cre3高度同源的序列Rccn4,设计出三条嵌套引物,采用SON-PCR(single oligonucleotide nested PCR)方法,从E-10基因组DNA中得到一个长为1264 bp的扩增产物(命名为Rccn-L),测序比对结果显示,这一序列将Rccn4的3’端延伸了1209 bp,与抗禾谷孢囊线虫Cre3基因核苷酸同源性为86﹪，核苷酸编码区长1026 bp，含一个不完整的开放阅读框,一个终止密码子，没有起始密码子和内含子结构,编码一个342个氨基酸残基的蛋白质。该蛋白质等电点为5.19，分子量为38112.6Da。从序列的第113位开始到第332位是NBS-LRR类抗病性基因LRR区,呈现XXLXXLXXL重复。LRR编码区内亮氨酸残基的含量达17﹪，与抗禾谷孢囊线虫Cre3基因LRR编码区的核苷酸和氨基酸同源性分别为89﹪和78﹪。本实验首次将SON-PCR成功地运用于植物基因克隆，为植物基因克隆提供了又一有效方法。 此外,还根据Cre3基因及其他的NBS-LRR类植物抗性基因的NBS和LRR区保守序列设计了两对特异性引物，从禾谷孢囊线虫抗性材料易变山羊草基因组DNA中扩增到两个相应的目标条带。测序分析结果表明，它们的长度分别为532bp和1175bp，构成了一个有32bp的共同序列的NBS-LRR编码区。其序列总长为1675bp（命名为RCCN），含有一个不完整的开放阅读框，没有起始密码子、终止密码子和内含子结构。其中编码序列为1673bp，可编码一个557个氨基酸的蛋白质，等电点（pI）为5.39，分子量为63537.5Da。与Cre3的核苷酸和氨基酸同源性分别为87.8﹪和77﹪。RCCN氨基酸序列中含有已知抗病基因NBS区域的几个保守模体：kinase2区的ILDD、kinase3的（ⅰ）ESKILVTTRSK，（ⅱ）KGSPLAARTVGG，（ⅲ）RRCFAYCS及EGF。RCCN NBS区与Cre3 NBS区的核苷酸和氨基酸的同源性分别为96.4﹪和94﹪。从氨基酸序列的274位到548位为LRR保守区，呈现不规则的aXXLXXLXXL（其中a代表I，V，L，F或M）重复，其中亮氨酸的含量为15.6﹪。该区域与Cre3的LRR区的核苷酸和氨基酸同源性分别为80.8﹪和74﹪。推测该序列可能为一个抗禾谷孢囊线虫的新基因。 本文对抗禾谷孢囊线虫基因的克隆研究，为进一步克隆基因全序列，探索其结构与功能，和研究该基因表达与调控提供了关键信息。同时也为通过基因工程途径将抗性基因向优良小麦品种高效、定向转移，最终培育出小麦抗禾谷孢囊线虫新品种奠定了基础。 Cereal cyst nematode (CCN) is a damaging pathogen of broad acre cereal crops in Australia, Europe, North America, India and China. It affects wheat, barley, oat and triticale and causes yield loss of up to 80%. At present, Transferring resistance genes against CCN into wheat cultivars and breeding varieties are considered one of the most effective methods for controlling the CCN. However, there are very limited reports concerning the cloning studies of resistance genes against the cereal cyst nematode. According to the sequence of Rccn4 which had high similarity to the nucleotide binding site (NBS) coding region of cereal cyst nematode resistance gene, Cre3， We designed three 3’ nested primers. Using single oligonucleotide nested PCR (SON-PCR) we successfully amplified one band, Rccn-L, of 1264bp from E-10 which is the wheat-Ae.variabilis translocation line containing the cereal cyst nematode resistance gene of Ae.variabilis. We found that this band of interesting is the 3’ flanking sequence of 1209bp in size of Rccn4. The coding region was 1026bp, which contained an incomplete open reading frame and a terminator codon, without initiation codon and intron, encoding a peptide of 342 amino acid residues, and shared 86﹪nucleotide sequence identity with Cre3. This peptide had a conserved LRR domain, containing the imperfect repeats,XXLXXLXXL, which contains 17﹪ leucine residues and shares, respectively, 89﹪ nucleotide sequence and 78﹪ amino acid sequence identity with the LRR sequence of Cre3 locus. This research firstly used SON-PCR in the research of plant genome successfully, which indicated that SON-PCR is another method of cloning plant gene. At the same time, According to the conversed motif of NBS and LRR region of cereal cyst nematode resistance gene Cre3 from wild wheat (Triticum tauschlii L.) and the known NBS-LRR group resistance genes, we designed two pairs of specific primers for NBS and LRR region respectively. One band of approximately 530bp was amplified using the specific primers for conversed NBS region and one band of approximately 1200bp was amplified with the specific primers for conversed LRR region. After sequencing, we found that these two sequences included 32bp common nucleotide sequence and have 1675 bp in total, which was registered as RCCN in the Genbank. RCCN contained a NBS-LRR domain and an incomplete open reading frame without initiation codon, terminator codon and inxon. Its exon encodes a peptide of 557 amino acid residues. The molecular weight of the protein from the amino acid was 63.537 KDa. The amino acid sequence of RCCN contained conserved motif: ILDD, ESKILVTTRSK, KGSPLAARTVGG, RRCFAYCS, EGF，LRR. RCCN shares 87.8﹪ nucleotide sequence and 77﹪ amino acid sequence identity with cereal cyst nematode gene Cre3. It might be a novel cereal cyst nematode resistance gene. These research results of cloning the resistance genes against cereal cyst nematode bring a great promise for transferring resistance genes into wheat cultivars and breeding new wheat varieties against cereal cyst nematode by gene engineering. And these results also lay the hard foundation for the expressing researches of these genes.

eta-production on the proton via electromagnetic and hadronic probes

The reactions pi(-)p -> eta n and gamma p -> eta p are investigated within a dynamical coupled-channels model of meson production reactions in the nucleon resonance region The meson-baryon channels included are pi N, pi Delta, sigma N, and rho N The direct eta-photoproduction process is studied within a formalism based on a chiral constituent quark model approach, complemented with a one-gluon-exchange mechanism, to take into account the breakdown of the SU(6)circle times O(3) symmetry In the models search, the following known nucleon resonances are embodied S-11(1535), S-11(1650), P-11(1440), P-11(1710), P-13(1720), D-13(1520), D-13(1700), D-15(1675), and F-15(1680). Data for the pi(-)p -> eta n reaction from threshold up to a total center-of-mass energy of W approximate to 2 GeV are satisfactorily reproduced For the photoproduction channel: two additional higher mass known resonances, P-13(1900) and F-15(2000), are also considered However, reproducing the data for gamma p -> eta p requires, within our approach, two new nucleon resonances, for which we extract, mass and width

Preliminary study of a niobium quarter-wave prototype cavity for a heavy-ion superconducting linac

Superconducting quarter-wave resonators, due to their compactness and their convenient shape for tuning and coupling, are very attractive for low-beta beam acceleration. In this paper, two types of cavities with different geometry have been numerically simulated: the first type with larger capacitive load in the beam line and the second type of lollipop-shape for 100 MHz, beta=0.06 beams; then the relative electromagnetic parameters and geometric sizes have been compared. It is found that the second type, whose structural design is optimized with the conical stem and shaping drift-tube, can support the better accelerating performance. At the end of the paper, some structural deformation effects on frequency shifts and appropriate solutions have been discussed.

番茄专用肥肥效初步研究

番茄专用肥肥效初步研究刘子江，崔桂霞，武冠云（中国科学院沈阳应用生态研究所）赵彩霞，张玉兴王述麟，王阳（沈阳市于洪区畜牧副食局）（沈阳市东陵区畜牧副食局）随着农业生产的发展和科学施肥技术的实施，专用肥的应用愈来愈广泛。专用肥的使用效果，是肥料研究领域...

黄土高原南部3种农田土壤剖面坚实度的变化规律

为了揭示黄土高原南部地区不同质地类型土壤剖面坚实度的变化及其与土壤含水率的定量关系,以黄墡土、土娄土、裸露在地表的粘化层耕作剖面为研究对象,定位观测其0~45 cm土壤坚实度与含水率的变化。结果表明,黄墡土、土娄土、裸露在地表粘化层耕作剖面的犁底层平均坚实度均大于耕层,犁底层平均坚实度较耕层分别高194.8%,87.3%,10.4%;剖面土壤质地越粘其平均坚实度越大;土壤坚实度与含水率呈负相关关系;土壤坚实度变化速率为0时,以上3种土壤剖面临界含水率分别为0.1712,0.1757,0.1835;质地不同的土壤剖面坚实度时空变化特征有差异,其中黄墡土剖面0~20 cm土层土壤坚实度为350~500 kPa,受土壤含水率变化的影响较小;20~30 cm土层土壤的坚实度为500~1400 kPa,不易受外界环境影响;30 cm以下土层土壤坚实度为700~1600 kPa,受土壤含水率变化影响较大。土娄土剖面0~40 cm土层土壤坚实度为600~1200 kPa,受含水率变化影响较大;40 cm以下土层土壤坚实度稳定在1 800 kPa左右。粘化层剖面0~15 cm土层土壤坚实度在2000 kPa左右,受环境影响较...

维生素C“二步发酵法”第二步发酵混合菌关系的研究

维生素Ｃ“二步发酵法”第二步发酵混合菌关系的研究马成新，杨凤玫，焦鹏，薛德林，王书锦（中国科学院沈阳应用生态研究所，１１００１５）维生素Ｃ作为一种重要的医药产品，不但用于治疗多种疾病，而且已被广泛用于食品、化妆品、饲料产品中，随着维生素Ｃ应用范围的增...

Synthesis and Upconversion Luminescence Properties of GdF3:Er3+, Yb3+

GdF3:Er3+,Yb-3 with Er3+ ion of 3% and Yb3+ ion concentration of 10%, 20% have been prepared by a hydrothermal method. The results of XRD show that all the samples are of an orthorhombic structure. The average crystallite sizes estimated by Scherrer formula are 28 and 26 nm for Gd0.87Yb0.10Er0.03F3 and Gd0.77Yb0.20Er0.03F3, respectively. The Upconversion luminescence spectra of the samples have been studied under 980 run laser excitation. The results show that the green and red upconversion emission can be attributed to the H-2(11/2),S-4(3/2) -> 4I(15/2) and 4F(9/2) -> 4I(15/2) transitions of Er3+, respectively.

7-Amino actinomycin D bound to single-stranded hairpin DNA enhanced by loop sequence-dependent luminescent Eu3+ and Tb3+ binding

Lanthanide Eu3+ and Tb3+ ions have been widely used in luminescent resonance energy transfer (LRET) for bioassays to study metal binding microenvironments. We report here that Eu3+ or Tb3+ can increase the binding affinity of antitumor antibiotic drug agent, 7-amino actinomycin D (7AACTD), binding to 5'-GT/TG-5' or 5'-GA/AG-5' mismatched stem region of the single-stranded hairpin DNA. Further studies indicate that the effect of Eu3+ or Tb3+ on 7AACTD binding is related to DNA loop sequence. Our results will provide new insights into how metal ions can enhance antitumor agents binding to their targets.

Synthesis and characterization of white-light-emitting polyfluorenes containing orange phosphorescent moieties in the side chain

Two orange phosphorescent iridium complex monomers, 9-hexyl-9-(iridium (III)bis(2-(4'-fluorophenyl)-4-phenylquinoline-N, C-2')(tetradecanedionate-11,13))-2,7-dibromofluorene (Br-PIr) and 9-hexyl-9-(iridium(III)bis(2-(4'-fluorophenyl)-4-methylquinoline-N, C-2')(tetradecanedionate-11,13))-2,7-dibromofluorene (Br-MIr), were successfully synthesized. The Suzuki polycondensation of 2,7-bis(trimethylene boronate)-9,9-dioctylfluorene with 2,7-dibromo-9,9-dioetylfluorene and Br-Plr or Br-MIr afforded two series of copolymers, PIrPFs and MIrPFs, in good yields, in which the concentrations of the phosphorescent moieties were kept small (0.5-3 mol % feed ratio) to realize incomplete energy transfer. The photoluminescence (PL) of the copolymers showed blue- and orange-emission peaks. A white-light-emitting diode with a configuration of indium tin oxide/poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate)/PIr05PF (0.5 mol % feed ratio of Br-PIr)/Ca/Al exhibited a luminous efficiency of 4.49 cd/A and a power efficiency of 2.35 lm/W at 6.0 V with Commission Internationale de L'Eclairage (CIE) coordinates of (0.46, 0.33). The CIE coordinates were improved to (0.34, 0.33) when copolymer MIr10PF (1.0 mol % feed ratio of Br-MIr) was employed as the white-emissive layer. The strong orange emission in the electroluminescence spectra in comparison with PL for these kinds of polymers was attributed to the additional contribution of charge trapping in the phosphorescent dopants.