抗大麦黄矮病毒小麦——中间偃麦草二体异代换系的选育和细胞,生？…

结合花药培养和常规选育,从７７－５４３３＊中５杂交组合后代中选育出６个抗大麦黄矮病毒（ｂａｒｌｅｙ ｙｅｌｌｏｗ ｄｗａｒｒｆ ｖｉｒｕｓ,ＢＹＤＶ）、染色体数目为４２的小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍＬ．）新种质,并用减数分裂配对分析、单体小麦测交法、基因组原位杂交、Ｃ分带、同工酶等电聚焦和ＲＡＰＤ对上述材料进行了综合鉴定;表明这些材料都是小麦－中间偃麦草（Ａｇｒｏｐｙｒｏｎ ｉｎｔｅｒ     

小麦-中间偃麦草双体异附加系的选育和鉴定

在小麦-中间偃麦草59个杂交后代种质系中,筛选出6个小麦-中间偃麦草双体异附加系(line 0605,line 0607,line 0609,line 0610,line 06ll,line 0625),并对其进行了形态学、白粉病抗性、细胞学和RAPD鉴定。形态学结果表明：6个双体异附加系农艺性状较好地结合了双亲的优良特点;细胞学结果表明：6个双体异附加系具有高度的细胞学稳定性,花粉母细胞减数分裂中期I(PMCMI)的染色体构型为2n=22II;RAPD分析表明：在供试的209个随机引物中有5个引物分别能在6个异附加系中稳定地扩增出不同的特异带型,可以作为各个异附加系所附加染色体的特异分子标记;白粉病抗性鉴定结果表明：line 0605表现免疫,line 0610和line 0625表现高抗,line 0607表现中抗,line 0609和line 06ll表现中感。     

一个多抗小麦—中间偃麦草新种质的选育和细胞分子生物学鉴定

经过多年田间和温室接种抗病性鉴定,从(77-5433×中5)杂交组合花药培养后代中选育出一个兼抗大麦黄矮病、条锈、叶锈和秆锈4种小麦主要病害的新种质遗4212。遗4212的体细胞染色体数为42,在减数分裂中期Ⅰ,在几乎所有的花粉母细胞中都可以观察到21个二价体,这说明遗4212是一个在遗传上业已稳定的整倍体材料。对(遗4212×77-5433)F_1代花粉母细胞的观察表明,遗4212可能是含1对外源中间偃麦草染色体的代换系或具较大中间偃麦草染色体片段的易位系。用基因组原位杂交(genomic in situ hybridization,GISH)对遗4212的有丝分裂中期相、减数分裂后期Ⅰ相和(遗4212×77-5433)F_1代花粉母细胞减数分裂中期Ⅰ、后期Ⅰ进行了检测,确证遗4212含1对外源中间偃麦草染色体。这些结果表明,遗4212是一个小麦一中间偃麦草代换系,其抗病性来自其携带的1对中间偃麦草。     

小麦-中间偃麦草双体异附加系的选育和鉴定

在小麦-中间偃麦草59个杂交后代种质系中,筛选出6个小麦-中间偃麦草双体异附加系(line0605,line0607,line0609,line0610,line0611,line0625),并对其进行了形态学、白粉病抗性、细胞学和RAPD鉴定。形态学结果表明:6个双体异附加系农艺性状较好地结合了双亲的优良特点;细胞学结果表明:6个双体异附加系具有高度的细胞学稳定性,花粉母细胞减数分裂中期Ⅰ(PMCMI)的染色体构型为2n=22Ⅱ;RAPD分析表明:在供试的209个随机引物中有5个引物分别能在6个异附加系中稳定地扩增出不同的特异带型,可以作为各个异附加系所附加染色体的特异分子标记;白粉病抗性鉴定结果表明:line0605表现免疫,line0610和line0625表现高抗,line0607表现中抗,line0609和line0611表现中感。     

普通小麦-百萨偃麦草(Thinopyrum bessarabicum)二体异附加系的选育与鉴定

为转移与利用百萨偃麦草耐盐、抗病等优良基因,用普通小麦中国春-百萨偃麦草双倍体与中国春杂交,通过染色体C-分带、分子原位杂交并结合减数分裂中期I的染色体配对分析,从回交后代中选育出一套小麦-百萨偃麦草二体异附加系。对这套异附加系进行的鉴定与分析表明,各附加系除添加了一对百萨偃麦草染色体外,小麦的21对染色体未见明显变化。各附加系所添加的百萨偃麦草染色体在减数分裂中期I配对基本正常,仅有少量单价体,其自交后代中外源染色体亦能正常传递。这说明所培育的这套二体异附加系在细胞学上已相对稳定,暂分别编号为DAJ1、DAJ2、DAJ3、DAJ4、DAJ5、DAJ6和DAJ7。各异附加系中百萨偃麦草染色体在小麦族中的部分同源群归属和百萨偃麦草耐盐抗病基因在染色体上的定位研究正在进行之中。     

小麦—中间偃麦草双体异附加系的选育及形态学和细胞学鉴定研究

应用在小麦品种烟农15与中间偃麦草杂交的五个世代群体中直接筛选2n=22Ⅱ植株的方法,获得11个双体异附加株,分别命名为DAL1、DAL2、……、DAL11。双体异附加株的细胞学稳定性较强,外源染色体传递频率高。形态学和细胞学鉴定结果表明：DAL1、3、5、6、8、9、10、11等8个异附加系中附加的可能是中间偃麦草第2部分同源群的染色体。 其中,DAL5、9、10、11是同一种异附加系,DAL6和DAL8为同一种异附加系,DAL3可能与之相同;DAL1与上述7个异附加系均不同。DAL2和DAL4可能分别附加了中间偃麦草第5部分同源群的1对染色体,但二者在形态上存在差异。DAL7可能附加了中间偃麦草第7部分同源群的1对染色体。旗叶卷曲是异附加系DAL、3、5、6、8、9、10、11共有的形态标记。11个异附加系可作为进一步研究和转移中间偃麦草有益基因的良好中间材料。     

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 .     

小麦-黑麦二体异附加系分子细胞遗传学鉴定

用形态学、细胞学、生化及分子标记方法,对小麦-黑麦衍生系N9436-0-2-29-9进行分析鉴定,以明确N9436-0-2-29-9中具体的外源染色体。农艺性状调查表明,N9436-0-2-29-9衍生系在形态上整齐一致,对白粉病高抗;细胞学分析表明,染色体构型稳定,2n=44=22Ⅱ;Giemsa C-分带和SSR分析表明,该衍生系为普通小麦-黑麦6R二体异附加系;高分子量麦谷蛋白亚基分析(HMW-GS)表明,外源染色体的导入使该衍生系的高分子量麦谷蛋白亚基表达受到影响。     

Creation and Cytological,Biochemical, Molecular Identification of Alien Disomic Substitution Lines with BYDV-resistance from Triticum aestivum-Agropyron intermedium Hybrids

By the method of combined anther culture and conventional selection, six new BYI)V (barley yellow dwarf vims)-resistant wheat germplasms (2n = 42) were obtained from the crosses between common wheat cv. 77-5433 and Zhong 5, a partial Triticum aestivum-Agrotryron intermedium amphiploid. The six germplasms were comprehensively identified by meiosis pairing analysis, testing cross analysis using monosomic analysis, C-banding, isoelectrofocusing (IEF) of isoenzyme and RAPD analyses. The results indicated that all the germplasms were alien substitution lines carrying resistant genes originated from Zhong 5.     

一个小麦-中间偃麦草异代换系的形态学和细胞学鉴定

中间偃麦草含有丰富的优良基因,在小麦的遗传改良中具有重要利用价值。对从中间偃麦草与小麦品种烟农15杂种后代(BC2F4)中选育的小麦种质系山农0095进行形态学和细胞学鉴定,结果表明：山农0095株高78cm,穗长17．3cm,旗叶长36．3cm,旗叶宽3．03cm,茎杆粗壮,繁茂性好,既长又宽的旗叶、长圆锥型穗是其显著的形态学特征;其根尖细胞染色体数日为2n=42,花粉母细胞减数分裂中期Ⅰ(PMC M Ⅰ)染色体构型为2n=21Ⅱ;它与普通小麦的杂种FⅠPMC M Ⅰ绝大多数细胞出现2个单价体,没有观察到多价体,平均染色体构型为2n=20．08Ⅱ 1．84Ⅰ。以上结果表明,山农0095是一个小麦-中间偃麦草的双体异代换系。     

小麦-大麦异代换系的创制及鉴定研究

利用"缺体回交法"以小大麦二体异附加系WBA9816作父本与阿勃缺体小麦杂交,创制小大麦异代换系.F1再用该异附加系回交,回交后代通过细胞学鉴定,筛选2n=43的双单体植株套袋自交,从自交后代群体培育出WBS02126;用原位杂交GISH和染色体C-分带技术鉴定表明,WBS02126为2D/2H异代换系;田间试验结果显示,WBS02126生长发育良好,育性基本正常,表现弱春性,叶片宽厚上挺,叶色淡绿,棒状穗,小穗排列紧密,长芒,早熟,综合抗病性好.     

抗白粉病小麦-中间偃麦草二体异附加系与杀配子材料杂交后代的细胞遗传学研究

利用已选育的抗白粉病烟农15-中间偃麦草二体异附加系与农林26-离果山羊草3C染色体附加系杂交．对其F1、F2、F3的细胞遗传学进行研究．结果表明：F1花粉母细胞减数分裂中期Ⅰ染色体构型紊乱,在39．48％的细胞中发现染色体断片、单价体,后期Ⅰ、后期Ⅱ出现落后染色体、染色体桥．四分体期微核出现频率达48．65％,说明杀配子染色体可有效诱导染色体发生断裂等结构变异;F2代在细胞学方面仍不稳定,表现为染色体数目发生变异,花粉母细胞减数分裂染色体构型紊乱。多价体、落后染色体、染色体桥及微核的普遍出现．说明染色体问可能发生断裂、重接、交换或易位等现象,F2代白粉病抗性也出现分离;F3代虽然染色体数日和白粉病抗性仍在分离,但花粉母细胞减数分裂中期Ⅰ染色体构型较F2稳定,相对紊乱系数下降．从F3代鉴定出了染色体数目为42、构型稳定且对白粉病表现免疫的单株。     

Genomic in situ hybridization (GISH) analyses of Thinopyrum intermedium, its partial amphiploid Zhong 5, and disease-resistant derivatives in wheat

Genomic in situhybridization (GISH) to root-tip cells at mitotic metaphase, using genomic DNA probes from Thinopyrum intermedium and Pseudoroegneria strigosa, was used to examine the genomic constitution of Th. intermedium, the 56-chromosome partial amphiploid to wheat called Zhong 5 and disease-resistant derivatives of Zhong 5, in a wheat background. Evidence from GISH indicated that Th. intermedium contained seven pairs of St, seven J^S and 21 J chromosomes; three pairs of Th. intermedium chromosomes with satellites in their short arms belonging to the St, J, J genomes and homoeologous groups 1, 1, and 5 respectively. GISH results using different materials and different probes showed that seven pairs of added Th. intermedium chromosomes in Zhong 5 included three pairs of St chromosomes, two pairs of J^S chromosomes and two pairs of St-J^S reciprocal tanslocation chromosomes. A pair of chromosomes, which substituted a pair of wheat chromosomes in Yi 4212 and in HG 295 and was added to 21 pairs of wheat chromosomes in the disomic additions Z1, Z2 and Z6, conferred BYDV-resistance and was identical to a pair of St-J^S tanslocation chromosomes (StJ^S) in Zhong 5. The StJ^S chromosome had a special GISH signal pattern and could be easily distinguished from other added chromosomes in Zhong 5; it has not yet been possible to locate the BYDV-resistant gene(s) of this translocated chromosome either in the St chromosome portion belonging to homoeologous group 2 or in the J^S chromosome portion whose homoeologous group relationship is still uncertain. Among 22 chromosome pairs in disomic addition line Z3, the added chromosome pair had satellites and belonged to the St genome and homoeologous group 1. Disomic addition line Z4 carried a pair of added chromosomes which was composed of a group-7 J^S chromosome translocated with a wheat chromosome; this chromosome was different to 7 Ai-1, but was identical to 7 Ai-2. The leaf rust and stem rust resistance genes were located in the distal region of the long arm, whereas the stripe rust resistance gene(s) was located in the short arm or in the proximal region of the long arm of 7 Ai-2. A pair of J^S-wheat translocation chromosomes, which originated from the WJ^S chromosomes in Z4, was added to the disomic addition line Z5; the added chromosomes of Z5 carried leaf and stem rust resistance but not stripe rust resistance; Z5 is a potentially useful source for rust resistance genes in wheat breeding and for cloning these novel rust-resistant genes. GISH analysis using the St genome as a probe has proved advantageous in identifying alien Th. intermedium in wheat. Received: 17 May 1999 / Accepted: 22 June 1999     

OsBTF3过表达转基因水稻株系的创制及其分子鉴定

本研究旨在创制OsBTF3过表达转基因水稻株系,为验证OsBTF3基因在水稻抗性和生长发育中的功能、评价其在水稻农艺性状遗传改良中的应用价值提供试验材料。通过基因过表达载体构建、水稻愈伤组织诱导、农杆菌介导愈伤组织转化、植株再生、潮霉素抗性(Hyg^R)筛选及PCR验证、基因过表达RT-Q-PCR检测等方法,成功地获得了97个T₀代和20个T₁代过表达转基因株系,并分别得到分子验证。与野生型对照株相比,5个T₁代过表达株系中的OsBTF3基因表达水平显著提高,平均高达3.58倍。因此,由组成型表达的35S启动子驱动的OsBTF3基因在转基因水稻株系中成功地得到了增量表达,并对水稻生长发育、抗病性和抗逆性具有调控作用。     

两株滇产广义美味牛肝菌的分离培养及其分子鉴定

采用组织分离法从广义美味牛肝菌子实体分离获得2株稳定的菌株,初步研究了两菌株的分离和培养条件。用ITS序列分析,对分离菌株进行了分子鉴定。基于ITS序列构建的系统树表明两株菌属于美味牛肝菌复合群,并与夏生牛肝菌Boletus aestivalis（Paul.）Fr.有较近的亲缘关系。     

Development of Chromosome Segment Substitution Lines Derived from Backcross between Two Sequenced Rice Cultivars, <Emphasis Type="Italic">Indica</Emphasis> Recipient 93-11 and <Emphasis Type="Italic">Japonica</Emphasis> Donor Nipponbare

Chromosome segment substitution lines (CSSLs) are powerful tools for detecting and precisely mapping quantitative trait loci (QTLs) and evaluating gene action as a single factor. In this study, 103 CSSLs were produced using two sequenced rice cultivars: 93-11, an elite restorer indica cultivar as recipient, and Nipponbare, a japonica cultivar, as donor. Each CSSL carried a single chromosome substituted segment. The total length of the substituted segments in the CSSLs was 2,590.6 cM, which was 1.7 times of the rice genome. To evaluate the potential application of these CSSLs for QTL detection, phenotypic variations of seed shattering, grain length and grain width in 10 CSSLs were observed. Two QTLs for seed shattering and three for grain length and grain width were identified and mapped on rice chromosomes. The results demonstrate that CSSLs are excellent genetic materials for dissecting complex traits into a set of monogenic loci. These CSSLs are of great potential value for QTL mapping and plant marker-assisted breeding (MAB).