大麦DNA导入小麦诱导抗白粉病变异的研究

本试验利用外源DNA导入技术,研究了大麦DNA导入小麦品种后,D_1,D_2和D_3代的抗白粉病变异株(系)在不同条件下的抗性表现及产量构成、籽粒蛋白质和氨基酸含量的变化。试验结果表明,导入外源DNA的D_1代小麦檀株出。现了抗白粉病等多种变异类型,其中免疫和高抗白粉病变异株古2.77％,且抗性能够向子代传递,其D_2代在大田自然发病和温室接菌条件下,有5个株系抗性保持稳定,8个株系有分离,其中一个株系在D_2和D_3代抗性均稳定。在田间D_2代有2个稳定株系(D_2-20,D_2-29)的籽粒粗蛋白质含量,比受体分别高20.3％和15.76％,17种氨基酸总量分别高23.4％和27.5％。在温室这些性,状的数值也明显高于受体。     

离子束介导大豆DNA的小麦变异株系苗期酸性蛋白水解酶种类和活性分析

采用复性电泳技术对离子柬介导大豆DNA小麦变异株系苗期蛋白水解酶进行研究。结果表明：在1叶1心期,两变异株系都检测到了差异酶带,分别为267和214kD;2叶1心期,变异株系的酶带型与对照没有差异,只是活性的变化;3叶1心期,变异株系间、变异株系与对照的酶带出现很大的差异,酶带表现的活性也不同,个别酶带存在的时期发生了变化。     

离子束介导大豆DNA的小麦变异株系蛋白水解酶谱分析

为研究离子束介导大豆DNA的小麦变异株系在返青期、起身期和拔节期的蛋白水解酶变化．采用复性电泳对新乡9号和筛选出的高蛋白变异株系05—10-1和低蛋白变异株系05—6-1,做不同酸碱条件下的蛋白水解酶分析。结果显示．与对照相比。变异株系05-6—1在返青期酸性条件下少检测到一条29kD酶带;在起身期中性和碱性条件下少检测到一条49kD酶带：在拔节期碱性条件下多检测到一条154kD新酶带．而少检测到158kD的酶带：而变异株系05～10—1在这3个生理时期和对照的蛋白水解酶酶谱带型基本一致．主要在某些酶带的酶活上存在差异。     

离子束介导大豆DNA的小麦变异株系蛋自水解酶谮分析

为研究离子束介导大豆DNA的小麦变异株系在返青期、起身期和拔节期的蛋白水解酶变化,采用复性电泳对新乡9号和筛选出的高蛋白变异株系05-10-1和低蛋白变异株系05-6-1,做不同酸碱条件下的蛋白水解酶分析.结果显示.与对照相比.变异株系05-6-1在返青期酸性条件下少检测到一条29kD酶带:在起身期中性和碱性条件下少检测到一条49kD酶带:在拔节期碱性条件下多检测到一条154kD新酶带.而少检测到158kD的酶带:而变异株系05-10-1在这3个生理时期和对照的蛋白水解酶酶谱带型基本一致.主要在某些酶带的酶活上存在差异.     

抗白粉病莆大麦5号的选育与浅识

抗白粉病莆大麦５号的选育与浅识陈炳坤，黄金堂，郭媛贞，蒋文广，陈德录（福建省莆田市农业科学研究所３５１１４４）陈桂煌，王元泉（福建省莆田市种子公司，黄石镇农技站）１选育经过与特征特性我所１９８３年冬以抗病多棱大麦木石港３号为母本，与综合性状优良的二棱...     

小麦×玉米杂交后代的农艺性状变异

从５个组合的小麦×玉米杂交后代中筛选出４２个变异株系与其小麦亲本作比较，这些株系在株高、有效穗数、穗长、穗粒数、粒重、成熟期及抗病性等性状产生了不同程度的变异，且这些变异是双向性的；从这些变异株系中筛选出了一些植株较矮、高抗白粉病、农艺性状好的株系，可以作为优良的育种亲本材料。证明小麦×玉米杂交途径在小麦单倍体育种、品种改良和提高育种效率方面具有重要实用价值。     

中国大麦酯酶同工酶的多样性及其地理分布研究

采用等电聚焦凝胶电泳，分析了５０２份栽培大麦和２３份西藏野生大麦，结果如下：（１）栽培大麦共出现４４条谱带，分１５组４５种带型，西藏野生大麦共７种带型；（２）酯酶同工酶的地理分布表明，具有相同或相近带型的品种分布与大麦生态区划和基因传播关系密切；（３）大多数西藏栽培大麦品种都有与西藏野生大麦相同的谱带，这条带除了在云南栽培大麦中发现外，其他栽培大麦品种中没有发现     

茄子抗冷细胞变异体的RAPD分析及自交一代株系的抗冷性鉴定

以茄子品系E－9903花药低温胁迫培养获得的抗冷细菌变异体为试材，用SDS方法从茄子幼嫩叶片提取DNA，选用40个长度为10个碱基的寡聚核苷酸（OPK01－OPK20，OPJ01－OPJ20）作为RAPD引物进行PCR扩增反应，并测定抗冷细胞变异体自交一代株系的冷害指数。结果表明：以OPK12为PCR反应引物时RAPD扩增谱带在抗冷性植株与同品系的对照株之间存在多态性差异，抗冷细胞变异体自交一代株系的冷害指数显著低于同品系的对照。     

小麦白粉病抗性QTL分析

本研究以国际小麦作图组织提供的(W7984×Opata85)重组近交群体为材料,2001－2002年对其亲本和114个株系进行了人工接种条件下的抗白粉病鉴定,并利用基于混合线性模型的复合区间作图软件QTLMAPER,进行了抗白粉病QTL分析,共检测到3个与小麦白粉病抗性相关的加性QTL,分别位于3B、5D、7D染色体上,可以解释42.8%的表型变异     

小麦抗白粉病生化标记——过氧化物酶pI6.1酶带

采用等电聚焦凝电泳比较了抗、感白现任小麦品种的过氧化物酶同工酶酶谱,发现抗、感品种之间的叶片过氧化物酶曲酶ＰＩ６．１酶带在拔节期间至白粉病发生期表现水平有很大差别。抗病品种中编码ＰＩ６．１酶带基因终生正常表达,该酶带在小麦全生育期均有较强的活性,染色后着色深,多属一级酶带。而该基因的表达在感病品种按节之后受到抑制,酶带变得模糊不清甚至消失踪迹,多为三级、零级酶带。感染白粉病之后感活性才回升,再度表     

Development of RFLP Markers for Barley

A PstI-based genomic library from barley DNA was screened for RFLPs in the three relatively-distant cultivars ‘Alexis’ (2-row spring type), ‘Igri’ (2-row winter type) and ‘Mammut’ (6-row winter type), digested with BamHI, EcoRI and HindIII. 50 % of the 108 DNA fragments studied represented single-copy sequences, 29 % low-copy and 21 % repetitive sequences. The DNA probes were assigned to discrete barley chromosomes with the aid of wheat/barley addition lines. 80 % of the single- and low-copy sequences hybridized with both barley and wheat DNA, whereas most repetitive sequences gave signals only with barley DNA.     

普通小麦品种“豫麦66”抗白粉病基因的鉴定与分子标记

豫麦66是对小麦白粉病(Blumeria graminis f. sp. tritici)具有良好抗性的小黑麦后代品种。本试验通过抗病鉴定与遗传分析, 明确了豫麦66携带1个抗白粉病显性单基因, 暂命名为PmYm66。采用2个以豫麦66为抗病亲本的杂交组合(豫麦66/铭贤169和豫麦66/ND3509)F2代抗、感病分离群体和F3代家系, 利用集群分离分析法(BSA)找到了与PmYm66连锁的分子标记XKsum193、EST48、EST83和EST84, 抗病基因和分子标记的顺序为EST48—EST83 (EST84)—Xksum193—PmYm66, 并通过中国春缺体-四体、双端体和缺失系将PmYm66基因及其连锁的分子标记定位在2AL染色体臂末端。多小种鉴定结果表明PmYm66(豫麦66)与2AL染色体臂上已有的Pm4a(Khapli/8Cc)和Pm4b(Armada)基因存在致病反应型差异。     

抗穗发芽2D/2H小大麦易位系的鉴定和应用研究

大麦2H染色体长臂上的Isa-H基因控制α-淀粉酶抑制蛋白的合成，减轻高α-淀粉酶活性对小麦穗发芽的不利影响。为了检测小大麦杂交后代中有无大麦Isa-H基因导入，利用染色体C-分带和原位杂交技术相结合对所创制的2H小大麦异附加系WBA9812和易位系WBT371进行了鉴定，以完整麦穗吸水保湿发芽法测定穗发芽的抗性，结合农艺性状观测，选育出具有抗穗发芽等优异特性的小大麦新种质。分析结果表明：WBT371是2D／2H易位系，抗小麦穗发芽。WBT371为进一步培育抗穗发芽小麦新品种创造了宝贵的种质资源。     

小麦背景下冰草6P染色体特异EST标记的开发

小麦-冰草[Agropyron cristatum (L. ) Gaertn.] 6P附加系普冰4844-12具有多粒等可用于小麦改良的优异基因。为高效率检测由小麦-冰草6P附加系衍生的易位系和渐渗系,以普冰4844-12及其亲本普通小麦Fukuhokomugi和冰草Z559 (2n=4x=28, PPPP)为材料,通过对冰草转录组测序获得的EST序列设计的P基因组EST分子标记引物,筛选在普通小麦背景下的6P染色体特异分子标记。结果从1 453对P基因组EST引物中筛选出130对小麦-冰草6P附加系特异标记引物。进而将这些特异标记与NCBI nr蛋白质数据库及小麦EST序列进行了比对,发现4条冰草EST序列的功能注释与抗病、抗逆相关;36条冰草EST与已经定位的小麦EST具有较高的相似性,其中33条(91.67%)位于小麦第6部分同源群染色体。为了进一步验证这些标记的特异性,分别对其中4个具有功能注释的EST标记在中国春等7个普通小麦背景下和随机选择的5个标记在小麦-冰草6P易位系背景下进行了检测,结果证明其确实可用于检测6P染色体。这些冰草6P染色体特异标记的开发为大规模地鉴定小麦-冰草衍生后代中P染色质成分奠定了基础。     

稀土元素对小麦芽鞘生长及内源吲哚乙酸含量的影响

本文研究了稀土元素对小麦芽鞘生长,吲哚乙酸氧化酶活性,过氧化物酶同工酶,内源吲哚乙酸水平的影响.结果表明:100ppm—1000ppm的硝酸稀土可促进小麦芽鞘的生长,尤以1000ppm浓度明显.这种生长的促进伴随着吲哚乙酸氧化酶活性的增加,过氧化物酶同工酶带数目的增多及内源吲哚乙酸水平的下降.稀土元素也可加强外源吲哚乙酸对小麦芽鞘生长的促进,抵消外源脱落酸对芽鞘生长的抑制.     

Cloning, genetic and physical mapping of resistance gene analogs in barley (Hordeum vulgare L.)

The majority of verified plant disease resistance genes isolated to date belong to the NBS‐LRR class, encoding proteins with a predicted nucleotide binding site (NBS) and a leucine‐rich repeat (LRR) region. Using degenerate primers, designed from the conserved motifs of the NBS region in tobacco N and Arabidopsis RPS2 genes, we isolated 190 resistance gene analogs (RGA) clones from barley genomic DNA. A total of 13 single‐ and low‐copy RGAs were genetically mapped onto chromosomes 1H–7H (except 5H) using three barley double haploid (DH) mapping populations: Steptoe × Morex, Harrington × TR306 and LUGC × Bowman. Sequence analysis of the RGAs showed that they are members of a diverse group. As a result of BLAST searches, one RGA proved unique as it did not detect any significant hit. Another RGA is putatively functional, because it detected several barley expressed sequence tag (EST) matches. To physically map the RGAs, 13 sequences were used to screen a 6.3 × cv. ‘Morex’ bacterial artificial chromosome (BAC) library. After fingerprint analysis, eight contigs were constructed incorporating 62 BAC clones. These BAC contigs are of great value for positional cloning of disease resistance genes, because they span the regions where various barley R genes have been genetically mapped.     

青稞HvBADH1基因的克隆及其转化烟草的初步研究

应用RT-PCR结合RACE技术, 从青稞总RNA中扩增得长度为1 512 bp的甜菜碱醛脱氢酶(BADH)基因cDNA全长编码序列。通过氨基酸同源性比对, 发现该序列的推定表达产物与大麦BADH同工酶BBD2的同源性为98.4%, 而与小麦、玉米和水稻等禾本科作物的BADH同源性分别为97.0%、84.7%和85.1%。将克隆到的青稞cDNA序列命名为HvBADH1, 投递到GenBank, 获得收录号EF492983。HvBADH1可以在原核表达系统TB1-pMAL c2x中正常表达出分子量为54.2 kD的多肽链。将HvBADH1的编码ORF, 插入到添加了植物表达CaMV 35S启动子和Nos polyA终止子调控元件的植物表达载体pCAMBIA1301质粒相应的克隆位点中, 构建了HvBADH1基因的农杆菌植物转化系统LBA4404(pCAM-ba)。进而采用农杆菌介导法, 将HvBADH1基因导入烟草中, 对所获得的潮霉素抗性烟草株系进行PCR、Southern blot和RT-PCR等分子生物学检测, 结果表明, 在得到的2个转基因株系中, HvBADH1基因已整合到受体植物基因组中, 并且可以在mRNA水平上进行转录。     

Identification of wheat–Thinopyrum intermedium 2Ai-2 ditelosomic addition and substitution lines with resistance to barley yellow dwarf virus

Among the regenerated plants derived from immature hybrid embryos of wheat–Thinopyrum intermedium disomic addition line Z6 × common wheat variety ‘Zhong8601’, a plant with a telocentric chromosome and barley yellow dwarf virus (BYDV) resistance was obtained. The telocentric chromosome paired with an entire Thinopyrum chromosome to form a heteromorphic bivalent at meiotic metaphase I. Genomic in situ hybridization showed that the telosome originated from Th. intermedium. Two ditelosomic additions and one disomic substitution were identified among the offspring of the plant. Two random amplified polymorphic DNA molecular markers were identified among 150 random primers used to detect the different arms of the alien chromosome. These might be useful for developing translocation lines with BYDV resistance.     

Comparison among available marker systems for cereal introgression breeding: A practical perspective

Summary There is an increasing amount of public sequence information for the main cultivated cereals, such as wheat and barley. It is not foreseeable that comparable efforts or resources could be devoted to related wild species. However, wild species are interesting sources of genetic variation through introgression breeding. Comparative genomics can be a helpful approach to make use of the available genomic resources. In this context, the potential of the wild barley species Hordeum chilense has been explored in recent years. It exhibits great levels of polymorphism and high crossability with different cereal genera. In addition, interesting biotic and abiotic stress resistance genes, and important quality traits like carotene content and seed storage protein variability shown in the species are also expressed in wheat backgrounds, and are the basis of a breeding program. Different approaches have been undertaken for tagging H. chilense genomic regions in a wheat background. The search for the most suitable DNA marker system started with the development of RAPD and SCAR markers due to a lack of sequence information from the wild species. Transferability of markers from wheat and barley (like STSs or SSRs) have also been useful approximations. More recently, SNP development is being accomplished for the species. In this work, the situation and prospects with the available molecular tools are considered from a practical point of view.