普通小麦-簇毛麦6VS/6AL易位系抗病性在不同小麦遗传背景中的传递

小麦条锈病和小麦白粉病是中国小麦的两大病害，由南京农业大学细胞遗传研究所培育的小麦-簇毛麦6VS／6AI。易位系高抗白粉病，并对中国当前新优势条锈菌生理小种表现高抗。用6VS／6AL易位系与中国不同小麦生产区的栽培品种宁春4号、扬麦5号、扬麦158、申32109、豫麦13、豫麦18等进行杂交，对其杂种后代进行田间抗条锈病和抗白粉病鉴定。从各杂交组合中均选出对白粉病和条锈病高抗的单株和株系，其抗病性在小麦不同遗传背景中可以正常表达。对从F3～F8代中选出的抗病材料进行根尖染色体C-分带和分子原位杂交鉴定，在所鉴定的高抗单株和株系中均含有一对或一条6VS／6AL易位染色体。     

Structural Changes of 2V Chromosome of Haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

Haynaldia villosa （2n=2X= 14, VV）, a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetic Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge （a remarkable morphology） mapped on 2V of Haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V（2D） and the hybrid F1 was then self-crossed. Chromosome C-banding, genomic in situ hybridization （GISH）, and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids Fz and F3. To date, four translocations including one small segmental translocation T6BS·6BL-2VS, two whole arm translocations （preliminarily designed as T3DS·2VL and T2VS.7DL） and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one isochromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25.120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 （2BL） tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS-7DL, Mt2VS, 2VS-2VS as well as the parent DS2V（2D）, whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C ofAegilops triuncialis could successfully induce chromosome 2V structural changes.     

iTRAQ-based quantitative proteomics analysis of defense responses triggered by the pathogen Rhizoctonia solani infection in rice

The soil-borne necrotrophic fungus Rhizoctonia solani is one of destructive fungi causing severe yield losses in various important crops. However, the host defense mechanisms against the invasion of this pathogen are poorly understood. In this study, we employed an i TRAQ-based quantitative proteomic approach to investigate host proteins responsive to R. solani using the resistant rice cultivar YSBR1. As a whole, we identified 319 differentially accumulated proteins(DAPs) after inoculation of ri...     

小麦—簇毛麦6VS／6AL易位系叶片cDNA文库构建及鉴定

用表达型质粒载体p^SPORT1构建了经白粉菌（Erysiphe graminis)诱导48h和未经诱导的小麦－簇毛麦6VS／6AL易位系（Triticum aestivum-Haynaldia villosa translocation lin)叶片cDNA库各一个，库宿主菌为E.coli DH10B，非诱导库包含约50万个重组cDNA克隆，平均插入征段为1．25kb，主要分布在400bp-2.2kb。诱导民含约30万个重组cDNA克隆，平均插入片段1．2kb。用克隆的病程相关基因（pathogenesis related gene)，＝－小麦类甜蛋白基因pWIR作探针，进行杂交筛库，杂交信号明显，重复性好。     

小麦—簇毛麦6VS/6AL易位材料幼胚一步成苗培养技术集成

 基因型差异是影响小麦幼胚一步成苗培养的主要因素。比较了小麦—簇毛麦易位材料与扬麦5号和中国春的一步成苗培养的污染率、成苗率、再生植株生根培养效率等主要技术指标,建立了适用于小麦—簇毛麦易位材料幼胚的—步成苗培养的有效技术体系,为培养纯合的普通小麦—簇毛麦6VS/6AL,小片段易位体系提供了技术支持。     

小麦一簇毛麦6VS／6AL易位材料幼胚一步成苗培养技术集成

基因型差异是影响小麦幼胚一步成苗培养的主要因素。比较了小麦一簇毛麦易位材料与扬麦5号和中国春的一步成苗培养的污染率、成苗率、再生植株生根培养效率等主要技术指标，建立了适用于小麦一簇毛麦易位材料幼胚的一步成苗培养的有效技术体系，为培养纯合的普通小麦一簇毛麦6VS／6AL，小片段易位体系提供了技术支持。     

普通小麦-簇毛麦6VS/6AL易位系cyclophilin基因的克隆与序列分析

利用RT-PCR的方法从普通小麦-簇毛麦6VS/6AL易位系克隆到1个cyp基因,命名为Ta-Hv-cyp。该基因全长为599 bp,编码1条171个氨基酸残基的多肽,具有保守的功能位点W128和胞质型CyP蛋白特有的插入序列KSGKPLH48~54。BLAST分析表明,推导的Ta-Hv-CyP蛋白分别与小麦、水稻、玉米、拟南芥的胞质型CyP具有98%,88%,85%和80%的氨基酸序列一致性。构建的进化树显示,推导的Ta-Hv-CyP蛋白与单子叶植物胞质型CyPs的亲缘关系较近。     

普通小麦-簇毛麦T5VS·5DL易位染色体对小麦主要农艺性状、品质和白粉病抗性的遗传效应分析

【目的】分析普通小麦-簇毛麦T5VS·5DL易位染色体对主要农艺性状、品质性状及白粉病抗性的遗传效应,了解T5VS·5DL易位染色体通过雌雄配子的传递情况,筛选便于育种利用的共显性分子标记,进一步明确T5VS·5DL易位系的高代回交品系在育种改良中的利用价值。【方法】分别以扬麦13、扬麦15为轮回亲本的高代T5VS·5DL易位系回交品系(BC4F4)及其分离群体(BC5F2)为材料,利用GISH及5VS特异分子标记对这些材料进行了鉴定;在大棚及大田2种环境下调查了这些材料的株高、穗长、小穗数、穗粒数、千粒重等主要农艺性状,并对这些材料的水溶剂保持力、碳酸钠溶剂保持力、蔗糖溶剂保持力、乳酸溶剂保持力、蛋白和籽粒硬度等品质性状进行了分析;还利用白粉菌混合生理小种对这些材料的苗期(二叶期)和成株期(抽穗期)进行了接种鉴定。【结果】含有T5VS·5DL易位染色体高代回交品系的株高、穗长、小穗数、穗粒数、千粒重等主要农艺性状与其轮回亲本相比,2种环境下的差异均不显著,表明簇毛麦5VS染色体臂代替普通小麦5DS染色体臂后,对产量性状的补偿性较好,没有显著的不利影响。品质性状的分析结果表明,含有T5VS·5DL易位染色体高代回交品系的籽粒硬度值(SKCS)均显著低于其轮回亲本,表明簇毛麦Dina/Dinb基因型比普通小麦的Pina/Pinb基因型具有更软质胚乳特性;另外,含有T5VS·5DL易位染色体高代回交品系的水溶剂保持力与碳酸钠溶剂保持力也显著低于轮回亲本,而蔗糖溶剂保持力、乳酸溶剂保持力及蛋白质含量的差异不显著,表明T5VS·5DL易位染色体对弱筋小麦品质指标可能有一定的正向效应。白粉菌混合生理小种接种鉴定的结果表明,在二叶期,含有T5VS·5DL易位染色体高代回交品系及其轮回亲本均高感白粉病,但在成株期含有T5VS·5DL易位染色体高代回交品系高抗白粉病,而其轮回亲本仍高感白粉病,表明含有T5VS·5DL易位染色体高代回交品系携带白粉病成株抗性基因。对BC5F2群体单株的白粉病及T5VS·5DL易位染色体鉴定的结果表明,所有含有T5VS·5DL易位染色体的单株均高抗白粉病,无T5VS·5DL易位染色体的单株均高感白粉病,推测一个显性的白粉病成株抗性基因与T5VS·5DL易位染色体共分离。同时在分离群体中,纯合易位染色体单株数、杂合单株数及无易位染色体单株数之比,经χ2测验符合1﹕2﹕1分离比例,表明T5VS·5DL易位染色体在小麦背景中遗传传递正常。通过对5VS特异分子标记的扩增条件及带型分析发现,共显性EST分子标记5EST-237、Pinb-1,扩增条件简单,特异带型易于识别,可以利用这些标记对T5VS·5DL易位染色体进行分子标记辅助选择。【结论】鉴于T5VS·5DL易位系良好的品质及抗病特性,建议在弱筋小麦的育种项目中加以利用。     

Development of EST-PCR Markers for the Chromosome 4V of Haynaldia villosa and Their Application in Identification of 4V Chromosome Structural Aberrants

EST-PCR based molecular markers specific for alien chromosomes are not only useful for the detection of the introgressed alien chromatin in the wheat background, but also provide evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers on chromosome 4V of Haynaldia villosa, a total of 607 primer pairs were designed according to the EST sequences, which were previously located in 23 different bins of wheat chromosomes 4A, 4B and 4D. By using the Triticum durum-H, villosa amphiploid and T. aestivum-H, villosa alien chromosome lines involving chromosome 4V, it was found that 9.23% of the tested primers could amplify specific bands for chromosome 4V. Thirty and twenty-six specific markers could be assigned to chromosome arms 4VS and 4VL, respectively. These 4V specific markers provided efficient tools for the characterization of structural variation involving the chromosome 4V as well as for the selection of useful genes located on chromosome 4V in breeding programs.     

小麦TaDREB6转录因子互作蛋白的筛选

 【目的】利用酵母双杂交技术从小麦cDNA文库中筛选TaDREB6互作蛋白,进一步研究DREB介导的小麦抗逆机制。【方法】以小麦的cDNA为模板扩增得到TaDREB6,构建小麦cDNA文库和pGBKT7-TaDREB6诱饵载体,将诱饵载体pGBKT7-TaDREB6以及pGADT7和cDNA文库混匀转入AH109酵母感受态细胞,在SD/-Ade/-His/-Leu/-Trp营养缺陷型培养基上,30℃培养3—5 d,挑取平板上直径大于2 mm的菌落,进行显蓝筛选。【结果】对筛选到的102个候选阳性克隆进行测序,在NCBI上进行BLAST比对分析,获得与能量代谢、抗逆和防御、转运、转录和翻译、信号转导以及生长发育等相关的候选蛋白。【结论】筛选得到候选蛋白的功能预测表明TaDREB6可能参与了多条信号转导途径,在抗逆境调控中具有重要作用。     

钾素水平对小麦氮素积累和运转及籽粒蛋白质形成的影响

 【目的】试图阐明施钾对小麦植株氮素积累、运转和籽粒蛋白质形成的影响机理。【方法】在池栽条件下，以宁麦9号（低蛋白）和扬麦10号（中蛋白）两个蛋白质含量不同的冬小麦品种为材料，研究了不同施钾水平下植株氮素积累、运转和开花期叶片含钾量的特征及其与籽粒蛋白质和各组分含量的关系。【结果】与不施钾相比，施钾提高了籽粒蛋白质含量，极显著提高了球蛋白和醇溶蛋白含量，由于对谷蛋白含量的作用甚微，因而显著降低了谷/醇比。施钾提高了开花期叶片含钾量进而显著促进了小麦植株花前氮素的积累和贮存氮素的运转，较高的开花期叶片钾营养水平显著提高了扬麦10号的花后氮积累，但对宁麦9号花后氮积累的促进作用较小。两种类型小麦的籽粒蛋白质含量对施钾的响应程度不同，扬麦10号大于宁麦9号。【结论】施钾条件下，因较高开花期叶片含钾量而显著提高的宁麦9号花前贮存氮素运转量和扬麦10号花后氮积累量，分别是这两种类型小麦籽粒蛋白质含量增加的重要生理原因。在本试验条件下，宁麦9号和扬麦10号的适宜施钾水平分别为K2O 150、225 kg·ha-1。     

簇毛麦抗白粉病基因向四川小麦转移的分子标记育种研究

利用小麦 6VS/6AL易位系的抗白粉病基因Pm2 1的SCAR标记 ,对不同簇毛麦来源的小麦抗白粉病材料及其杂交后代进行了分子鉴定和标记辅助选择研究 ,结果表明利用该标记可以鉴定小麦遗传背景下的簇毛麦 6VS染色体臂 ,而且引物对小麦背景中的多年生簇毛麦也同样得到扩增。对簇毛麦染色体易位系 6VS/6DL与四川小麦品种杂交的后代群体 ,可以利用SCAR标记进行辅助抗性选择。本文还对四川小麦分子标记辅助育种的策略进行了讨论     

普通小麦-簇毛麦易位系T6BS·6BL-2VS的选育(英文)

[Objective] The aim of experiment was to provide a new germplasm for wheat breeding by further using desirable genes in 2V chromosome of Haynaldia villosa.[Method] Through hybridization between common wheat(Triticum aestivum)-Haynaldia villosa disomic substitution line and common wheat Nonglin26-3C chromosome of Aegilops triuncialis disomic addition line,the analysis methods such as chromosome C-banding,genomic in situ hybridization and molecular marker technique were comprehensively applied and combined characters investigation.[Result] The wheat-Haynaldia villosa translocation line(T6BS·6BL-2VS)was selected from hybrid progenies to conduct characters investigation,which found some bristles on glume ridge of T6BS·6BL-2VS.[Conclusion] The translocation line induced by gametocidal chromosome was a small segment translocation line and the gene of bristle on glume ridge of Haynaldia villosa was located between the middle and the terminal of 2VS.     

玉米早期发育阶段粒位效应的蛋白质组学分析

【目的】从蛋白质组学的角度研究早期发育阶段玉米上、中部籽粒差异表达的蛋白质,分析其功能,探明玉米粒位发育差异的分子机理。【方法】在大田条件下,以粒位效应显著的玉米品种登海661（DH661）为供试材料,90 000株/hm²密度下种植,在开花期人工饱和授粉后0、3、6、12 d取果穗上部与中部籽粒。采用TCA-丙酮沉淀法提取籽粒总蛋白,双向电泳分离后获得蛋白质图谱。分别以0、3、6、12 d中部籽粒的凝胶作为参考胶,将上部籽粒凝胶与其进行比对,利用Image master 2D 7.0软件分析籽粒早期发育不同阶段上、中粒位蛋白质表达的差异。通过 MALDI-TOF/TOF MS 质谱分析及 NCBI数据库搜索,对差异表达蛋白质进行鉴定并分析其涉及的生物学功能。【结果】较高密度种植后,果穗籽粒早期发育阶段共检测到超过1000个清晰蛋白质点。通过图像处理软件成对匹配分析,果穗上、中部籽粒早期发育阶段差异蛋白质点为66个,其中52个蛋白质点与NCBI数据库匹配,鉴定率为78.8%。差异蛋白质涉及籽粒呼吸与能量代谢（10个蛋白质点,19%）、胁迫与防御（9个蛋白质点,17%）、蛋白质代谢（9个蛋白质点,17%）、氮代谢（6个蛋白质点,11%）、细胞分化与增殖（5个蛋白质点,10%）、转录与翻译（5个蛋白质点,10%）、次生物质代谢（3个蛋白质点,6%）等功能范畴。对相关的差异蛋白质表达丰度分析,与中部籽粒相比,上部籽粒涉及细胞分化与增殖、呼吸与能量代谢的大部分蛋白质在一个或多个时间段内均显著下调,说明上部籽粒胚乳细胞增殖及呼吸能量代谢能力显著降低。同时,上部籽粒涉及胁迫与防御的多种抗氧化酶系、乙二醛酶1以及涉及蛋白质代谢的4个分子伴侣蛋白质在发育早期也处于低水平表达,说明上部籽粒应对逆境条件防御能力较弱且蛋白质结构不稳定。另外,与中部籽粒相比,上部籽粒氮代谢中丙氨酸转氨酶以及S-腺苷甲硫氨酸合成酶1在授粉后6-12 d内均下调表达,说明上部籽粒氮同化能力较弱,影响后续的氨基酸合成与蛋白质代谢过程。【结论】与果穗中部籽粒相比,上部籽粒细胞分化与增殖相关蛋白质的表达水平较低,呼吸与能量代谢能力较弱,导致上部籽粒库容与库活性降低。另外,面对氧化应激等逆境时,上部籽粒相关的抗氧化酶以及分子伴侣蛋白表达水平较低,致使其防御能力低于中部籽粒。丙氨酸转氨酶、S-腺苷甲硫氨酸合成酶1（SAMS）的差异表达也可能是导致粒位效应的重要原因。     

小麦-簇毛麦端体附加系中6VS的细胞遗传学行为及其抗性遗传研究

小麦 -簇毛麦 6VS端体附加系AD134抗白粉病特性受 6VS上基因的控制 ,在不同小麦背景下都能充分表达。单体附加的 6VS抑制小麦染色体正常配对 ,造成单价体数量增加 ,但其影响程度还与小麦遗传背景的互作有关。观察到多种减数分裂异常现象 ,但染色体断裂的频率较低 ,表明使用端体附加系选育易位系应增大杂交后代的群体。AD134细胞学稳定性较差。在杂合状态下 ,6VS通过雌、雄配子的传递率均显著下降 ,但其通过雌配子的传递率 (平均 2 3 8% )仍显著高于通过雄配子的传递率 (平均 7 4% ) ,也高于 6V通过雌配子的传递率 ,文中对这一结果可能的原因进行了讨论     

Physiology and defense responses of wheat to the infestation of different cereal aphids

Cereal aphids are major insect pests of wheat, which cause significant damages to wheat production. Previous studies mainly focused on the resistance of different wheat varieties to one specific aphid species. However, reports on the physiology and defense responses of wheat to different cereal aphids are basically lacking. In this work, we studied the feeding behavior of three cereal aphids: the grain aphid, Sitobion avenae(Fabricius), the greenbugs, Schizaphis graminum(Rondani), and the bird cherry-oat aphid, Rhopalosiphum padi(Linnaeus) on winter wheat, and the physiology and defense responses of wheat to the infestation of these cereal aphids with focus on how these cereal aphids utilize divergent strategies to optimize their nutrition requirement from wheat leaves. Our results indicated that S. graminum and R. padi were better adapted to penetrating phloem tissue and to collect more nutrition than S. avenae. The harm on wheat physiology committed by S. graminum and R. padi was severer than that by S. avenae, through reducing chlorophyll concentration and interfering metabolism genes. Furthermore, cereal aphids manipulated the plant nutrition metabolism by increasing the relative concentration of major amino acids and percentage of essential amino acids. In addition, different cereal aphids triggered specific defense response in wheat. All of these results suggested that different cereal aphids utilize divergent strategies to change the physiological and defense responses of their host plants in order to optimize their nutrition absorption and requirement. These findings not only extend our current knowledge on the insect–plant interactions but also provide useful clues to develop novel biotechnological strategies for enhancing the resistance and tolerance of crop plants against phloem-feeding insects.     

抗白粉病小麦-簇毛麦易位系92R149的利用研究

以携带有抗白粉病基因Pm21的小麦-簇毛麦6VS/6AL易位系92R149为母本、大穗高感白粉病小麦品系忻96-74为父本,通过杂交进行抗病、丰产基因的重组,调查了亲本及F2,F3的几个主要农艺性状,并进行了变异系数分析。结果表明,92R149所特有的抗白粉病和忻96-74的大穗特性可直接传递给后代,且杂种后代的变异极为丰富,可选育出抗白粉病且具大穗的优异单株。     

3个小麦新种质的选育与鉴定

针对目前山西省小麦抗病性差的现状,以携带有抗病基因的小麦-簇毛麦6VS/6AL易位系92R137和92R149为亲本,与感病小麦品种忻96-74、矮多1号、黑粒小麦76杂交,进行抗病、丰产、优质和矮秆基因的重组,选育出3个抗病新种质G10,S06和S25,并综合应用农艺性状、细胞学、分子标记等方法对其进行鉴定。结果表明,G10为半矮秆、特大穗,对小麦白粉病和条锈病免疫;S06为矮秆,对白粉病免疫;S25具有5+10优质亚基、紫粒,对白粉病和条锈病免疫。G10,S06和S25是小麦抗病、优质育种的新种质材料。     

滁辐麦6号品种特性及栽培要点

[目的]介绍早熟、高产、抗病的小麦品种滁辐麦6号。[方法]对滁辐麦6号的产量、农艺性状、经济性状、抗逆性、品质及其栽培技术要点进行综述。[结果]滁辐麦6号是滁州市原子能利用研究所以麦优1号为母本、扬麦4号为父本进行杂交,杂交后的种子用Co60-γ辐照处理,再经系统选育而成。2005～2006年安徽省区试中,滁辐麦6号平均产量为6 493.5 kg/hm2,比对照扬麦158增产9.9%,2006～2007年安徽省生产试验中,滁辐麦6号平均产量为6 633.0 kg/hm2,比对照增产7.1%,是一个熟期适中、抗逆性强、适应性广、品质优良的品种,适宜在滁州市、淮南及长江中下游地区种植。[结论]该研究为滁辐麦6号的推广应用提供了理论依据。