抗条锈病小偃麦双体异附加系山农87074-519的鉴定

综合利用抗性接种鉴定、细胞学分析、SSR分子标记和基因组原位杂交(GISH)技术相结合的方法,对从长穗偃麦草与小麦复合杂交后代中选育的抗条锈病种质系山农87074-519进行了鉴定。结果表明,山农87074-519的根尖细胞染色体数目2n=44,花粉母细胞减数分裂中期I(PMCMI)绝大多数细胞内可观察到22个二价体,平均染色体构型2n=44=21.82Ⅱ 0.36Ⅰ,它与普通小麦中国春杂种F1的多数花粉母细胞内染色体构型为2n=21Ⅱ 1Ⅰ,因此它是1个附加了1对长穗偃麦草染色体的双体异附加系;以假鹅冠草St基因组总DNA作探针进行原位杂交发现山农87074-519的44条染色体中有2条出现黄绿色杂交信号,且杂交信号遍布整条染色体,证明其附加的长穗偃麦草染色体为St基组;利用SSR分子标记技术,在170对SSR引物中筛选出特异引物BARC165,它能稳定地在山农87074-519中扩增出长穗偃麦草特异标记BARC165268;将长穗偃麦草中BARC165的特异扩增片段克隆测序后制备成探针进行原位杂交,可在山农87074-519的间期染色体和有丝分裂中期染色体检测到杂交信号。山农87074-519综合农艺性状较好,对条锈病免疫,其抗性基因为显性,且位于附加的长穗偃麦草St基组染色体上,暂将其表示为YrSt。该种质系在小麦的遗传改良中具有重要利用价值。     

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。     

兼抗白粉和条锈病小滨麦种质系山农6343的细胞学和SSR鉴定

利用抗性接种鉴定、细胞学和SSR分子标记技术相结合的方法,对从八倍体小滨麦和普通小麦烟农15杂种后代选育出的兼抗白粉病和条锈病的小滨麦种质系山农6343进行了鉴定.结果表明,山农6343的根尖细胞染色体数目2n=42,花粉母细胞减数分裂中期I(PMC MI)绝大多数细胞内可观察到21个二价体,平均染色体构型为2n=21Ⅱ,与普通小麦烟农15杂种F1的花粉母细胞内观察到2n=19Ⅱ+1Ⅳ的染色体构型,四价体出现频率为24.1%.利用SSR分子标记技术,在1283对SSR和EST-SSR引物中筛选出两对特异引物BARC236-4A和KSUM134,均能稳定地在山农6343中扩增出滨麦草的特异标记BARC236255和KSUM134245,且两个标记在小滨麦易位系山农0096中得到了验证.初步确定山农6343是一个小滨麦易位系.由于在目前已命名的小麦白粉病和条锈病抗性基因中尚未有来自滨麦草的,推测山农6343可能为新的白粉病和条锈病抗源,对小麦白粉病和条锈病的抗性遗传改良将具有重要的利用价值.     

抗白粉病耐盐小麦-中间偃麦草附加系‘山农120211’的鉴定

以中间偃麦草(Thinopyrum intermedium,2n=42)与普通小麦‘烟农15’杂交,从其杂种后代中选育出一个细胞学稳定的二体异附加系‘山农120211’,该研究对其细胞学和主要性状特点进行了鉴定。白粉病抗性鉴定结果表明,‘山农120211’成株期对白粉病的田间抗性为免疫,苗期对白粉病菌种E09表现为免疫。以耐盐品种‘山融3号’为对照进行苗期耐盐性鉴定表明,‘山农120211’耐盐级别为2级(较强)。细胞学鉴定表明:‘山农120211’根尖细胞染色体数目为2n=44,PMC MI染色体构型为2n=22Ⅱ,具有高度的细胞学稳定性。以拟鹅观草基因组DNA为探针,‘烟农15’DNA为封阻,在‘山农120211’的根尖有丝分裂细胞中检测到2条染色体具有明显的杂交信号,确定其为二体异附加系。利用该实验室筛选的71对E组染色体特异分子标记,对‘山农120211’分析显示,标记BE494262在中间偃麦草和‘山农120211’中可以稳定扩增出1条440bp特异带,而‘烟农15’中缺少此带,BE494262可作为‘山农120211’中附加中间偃麦草染色体的特异标记。利用二倍体长穗偃麦草和一套中国春-长穗偃麦草异附加系(1Ee~7Ee),进一步将BE494262定位在2Ee染色体,确定‘山农120211’所附加的中间偃麦草染色体为2Ee染色体。     

兼抗白粉、条锈病小偃麦渗入系CH7124抗性遗传及细胞学鉴定

CH7124是通过八倍体小偃麦TAI8335与感病小麦杂交、回交育成的兼抗白粉病、条锈病的小偃麦种质系。利用抗性接种鉴定、细胞学和基因组原位杂交(GISH)技术相结合的方法,对CH7124的抗性来源、遗传方式及细胞学特征进行了分析和鉴定。结果表明,CH7124在苗期和成株期对条锈菌系CYR29、CYR31、CYR32、CYR33和白粉菌系E09、E20、E21、E26表现为免疫或近免疫,其抗性来自中间偃麦草,受1对显性核基因控制;CH7124的根尖细胞染色体数目为2n=42,花粉母细胞减数分裂中期I(PMC MI)绝大多数细胞内可观察到21个二价体,平均配对构型为2n=0.30 I+20.79 II+0.04 III;与普通小麦中国春、绵阳11的杂种F1中,有80%以上的花粉母细胞可观察到2n=21Ⅱ的染色体构型,其平均配对构型均为2n=21II。说明CH7124具有与普通小麦相似的染色体结构和规则的配对构型。由于利用以中间偃麦草总DNA为标记探针的原位杂交未观察到可见的外源DNA杂交信号,进一步证明CH7124是一个小麦-中间偃麦草的隐形异源渗入系。     

普通小麦与长穗偃麦草杂种及其衍生材料的细胞遗传学特性

对十倍体长穗偃麦草(Thinopyrum ponticum)与普通小麦杂交F1及其与普通小麦回交BC1F1的形态学和细胞学特性进行了分析。结果表明,长穗偃麦草与普通小麦‘兰考矮早八’衍生F1(‘兰考小偃麦’)的根尖细胞染色体数为56条;花粉母细胞减数分裂中期Ⅰ染色体构型平均值为19.81Ⅰ+15.78Ⅱ+0.75Ⅲ+0.59Ⅳ;基因组荧光原位杂交(GISH)显示,兰考小偃麦中含有35条完整的长穗偃麦草和21条小麦染色体。‘兰考小偃麦’/‘科育818’和‘兰考小偃麦’/‘Cp02-3-5-5’杂交F1的根尖细胞染色体数及其所遗传的长穗偃麦草染色体数分别为50~52和16~22条,且存在染色体易位;花粉母细胞减数分裂中期Ⅰ平均染色体构型为14.54Ⅰ+17.40Ⅱ+0.55Ⅲ+0.14Ⅳ,平均49.4%的细胞出现多价体(三价体或四价体)。这些材料为创造小麦-长穗偃麦草新种质奠定了基础。     

偃麦草属三个种的染色体组研究

本文综合应用体细胞染色体分带、杂交一代 PMc MI 染色体构型统计和同工酶分析,研究了偃麦草属三个种的染色体组构成。结果表明:中间偃麦草不含与小麦同源的 B 组,有两个组部分同源,其染色体组可用 xE_1E_2表示。长穗偃麦草(10x)也不含 B 组,有四个组两两部分同源,是部分同源异源十倍体,染色体组公式为 xE_1E_1F_1F_2。四倍体偃麦草的两个染色体组间缺少同源性。     

偃麦草与小偃麦染色体组构成的细胞遗传学研究在Ⅴ.硬粒小麦与中间偃麦草杂交及回交的细胞遗传学研究

获得了硬粒小麦(2n=6x=28、AABB)与中间偃麦草(2n=6x=42、NNE_1E_1E_2E_2)杂种F_1及回交后代材料。统计分析杂种F_1及回交一代PMC MI染色体配对构型,认为中间偃麦草具较远缘的同亲关系(distant homologous)染色体组。由三价体出现频率分析,中间偃麦草不含小麦的B染色体组,建议用NE_1E_2为其染色体组公式。根据回交一代及其自交后代染色体数目,分析了六倍体小偃麦这一人工新物种的形成过程。     

一个异源胞质单体附加系小麦的遗传分析

以长穗偃麦草(Elytrigia elongata=Agropyron elongatum,2n=70)为母本,普通小麦为父本,进行核代换回交,在第九次回交的F_1(BC,F_1)代里,发现了一个异源胞质单体附加系小麦,代号E617。以E617做母本,普通小麦做父本进行杂交,杂种F_1出现正常株和弱株两种类型。正常株具有43条染色体,弱株具有42条染色体。反交,以普通小麦做母本,E617做父本,杂种F_1,无论是具有43条染色体还是具有42条染色体的植株,都是正常的。正反杂交结果表明,长穗偃麦草细胞质对普通小麦的细胞核产生了抑制作用,使核质杂种生长不正常,而附加一条特异的长穗偃麦草染色体(或端体),就能消除长穗偃麦草细胞质的抑制作用,使核质杂种恢复正常生长。     

小麦-簇毛麦种质系‘山农030713＇的细胞学和SSR鉴定

利用形态学、细胞学以及SSR标记技术对从硬簇麦和Am3的杂种后代中选育的种质系‘山农030713＇进行了鉴定,结果表明：种质系‘山农030713＇大田生长整齐一致,农艺性状较好,且对白粉病免疫;其根尖细胞染色体数目为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F1PMC MⅠ多数细胞中形成21个二价体,且常有四价体出现,可能伴有染色体的结构变异;SSR分析证明‘山农030713＇基本染色体组成为AABBDD,引物Xgwm99-1A在‘山农030713＇中扩增出簇毛麦的特异带,表明‘山农030713＇中有来自于簇毛麦的遗传物质,此特异带可作为识别‘山农030713＇的SSR标记.综合形态学、细胞学和SSR分析结果推测,‘山农030713＇可能是一个小麦-簇毛麦易位系.     

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。     

抗白粉病基因PmCH1302的遗传分析及染色体定位

CH1302是以来源于中间偃麦草的八倍体小偃麦TAI7047为桥梁亲本选育的高抗白粉病的小麦新品系,对白粉菌多个流行小种均表现出良好抗性。为了解其抗白粉病基因来源及其在染色体上的位置,对绵阳11×CH1302的F_1、F_2及F_(2∶3)家系进行了遗传分析,推断其抗白粉病基因可能来源于中间偃麦草,暂将其命名为PmCH1302。利用i Select 90K SNP芯片对抗、感病池进行扫描,发现位于2AL染色体上的多态性位点最多,为313个,占全部多态性位点的9.79%,且集中于2AL染色体100~105 c M和150~155 cM两个区域附近。在上述位点选取SSR标记,筛选出3对与Pm CH1302连锁的分子标记,Xwmc522、Xgwm356和Xgwm526,其中Xgwm356和Xgwm526位于Pm CH1302两侧,连锁距离分别为3.1 c M和7.8 cM。利用遗传图谱以及中国春缺体、双端体将PmCH1302定位于小麦2AL染色体上。进一步与位于2AL上的Pm4、Pm50比较发现,PmCH1302可能是位于2AL上的一个新基因或等位基因。     

Powdery Mildew Resistance in Barley Landraces from Morocco

Nineteen barley landraces collected from Morocco were screened for resistance to powdery mildew. The landraces originated from the collection at the Polish Gene Bank, IHAR Radzików, Poland. The fifteen landraces tested showed powdery mildew resistance reactions and 35 single plant lines were selected. Twenty-one of these lines were tested in the seedling stage with 30, four lines with 17 and another 10 lines with 23 differential isolates of powdery mildew, respectively. The isolates were chosen according to their virulence spectra observed on the Pallas isolines differential set. Nine lines (E 1029-1-1, E 1042-2-2, E 1050-1-1, E 1054-5-1, E 1056-2-5, E 1056-3-1, E 1061-1-1, E 1061-1-3 and E 1067-1-2) which originated from seven landraces showed resistance to all prevalent European powdery mildew virulence genes. The most frequent score was 2 and 16 lines showed this reaction for inoculation with most isolates used. The distribution of reaction type indicated that about 77% of all reaction types observed were classified as powdery mildew resistance (scores 0, 1 and 2). In all lines the presence of unknown genes alone or in combinations with specific ones was postulated. Four different resistance alleles ( Mlat , Mla6 , Mla14 and Mla12 ) were postulated to be present in 10 tested lines alone or in combination. Alleles Mlat , Mla6 and Mla14 were postulated to be present in four and Mla12 in two tested lines, respectively. The value of barley landraces for diversification of resistance genes for powdery mildew is discussed.     

RAPDs as molecular markers for the detection of Aegilops markgrafii chromatin in addition and euploid introgression lines of hexaploid wheat

 Aegilops markgrafii contains resistance genes to powdery mildew, leaf rust and stripe rust, and also has high crude protein and lysine contents, which can be useful for wheat improvement. These important traits are localized on different chromosomes. Disomic Triticum aestivum-Ae. markgrafii addition lines and euploid introgression lines showing leaf-rust and powdery mildew resistance were screened with RAPDs to detect chromosome-specific markers which can accelerate the breeding process. RAPD markers for all six available disomic addition lines were obtained. The additional chromosomes B, C, D, E, F and G were identified by three, three, three, two, one and seven primers, respectively. All three chromosome-B-specific RAPD markers demonstrated the presence of alien chromatin in the leaf-rust-resistant 42-chromosome introgression lines as well as in the segregating progeny. The three chromosome-C-identifying primers also demonstrated the presence of that chromosome in powdery mildew-resistant euploid introgression lines. The substitution lines (5A)5C and (5D)5C with different genetic backgrounds for both parents, in comparison to the lines mentioned above, showed the chromosome C-specific band with only two of the three primers. The chromosome F-specific primer and a primer evident on all the Ae. markgrafii chromosomes analysed did not generate the expected fragments on the chromosome F_del addition line, indicating that the markers are located on the deleted part of chromosome F. Received: 20 August 1996 / Accepted 17 January 1997     

Introgression of Resistance to Powdery Mildew Conferred by Chromosome 2R by Crossing Wheat Nullisomic 2D with Rye

Using the nulUsomic back-cross procedure, four wheat-rye chromosome substitution 2R （2D） lines with different agronomic performance, designated WR02-145-1, WR01-145-2, WR02-145-3, and WR02-145-4, were produced from a cross between 2D nullisomic wheat （Triticum aestivum L. cv. ＂Xiaoyan 6＂） and rye （Secale cereale L. cv. ＂German White＂）. The chromosomal constitution of 2n=42=21 in WR02-145 lines was confirmed by cytological and molecular cytogenetic methods. Using genomic in situ hybridization on root tip chromosome preparations, a pair of intact rye chromosomes was detected in the WR02-145 lines. PCR using chromosome-specific primers confirmed the presence of 2R chromosomes of rye in these wheat-rye lines, indicating that WR02o145 lines are disomic chromosome substitution lines 2R （2D）. The WR02-145 lines are resistant to the powdery mildew （Erysiphe graminis DC. f. sp. tritici E. Marchal） isolates prevalent in northern China and may possess gene（s） for resistance to powdery mildew, which differ from the previously identified Pm7gene located on chromosome 2RL. The newly developed ＂Xiaoyan 6＂- ＂German White＂ 2R （2D） chromosome substitution lines are genetically stable, show desirable agronomic traits, and are expected to be useful in wheat improvement.     

小麦白粉病抗性基因的导入及AFLP分析

本研究以簇毛麦为抗源,采用杂交与辐射、组织培养相结合的方法,将簇毛麦的抗白粉病基因导入小麦,选育出高产、抗白粉病的小麦新品种和农艺性状较好、抗白粉病的小麦新种质。经AFLP分析,确定4个抗白粉病种质均为含有一段簇毛麦DNA的易位系。并得到3个可能与抗性基因紧密连锁的标记。     

Molecular cytogenetic characterization of wheat--Thinopyrum elongatum addition, substitution and translocation lines with a novel source of resistance to wheat Fusarium Head Blight

Thinopyrum elongatum(2n = 2x = 14,EE),a wild relative of wheat,has been suggested as a potentially novel source of resistance to several major wheat diseases including Fusarium Head Blight(FHB).In this study,a series of wheat(cv.Chinese Spring,CS) substitution and ditelosomic lines,including Th.elongatum additions,were assessed for TypeⅡresistance to FHB.Results indicated that the lines containing chromosome 7E of Th.elongatum gave a high level of resistance to FHB,wherein the infection did not spread beyond the inoculated floret.Furthermore,it was determined that the novel resistance gene(s) of 7E was located on the short-arm(7ES) based on sharp difference in FHB resistance between the two 7E ditelosomic lines for each arm.On the other hand,Th.elongatum chromosomes 5E and 6E likely contain gene(s) for susceptibility to FHB because the disease spreads rapidly within the inoculated spikes of these lines. Genomic in situ hybridization(GISH) analysis revealed that the alien chromosomes in the addition and substitution lines were intact,and the lines did not contain discernible genomic aberrations.GISH and multicolor-GISH analyses were further performed on three translocation lines that also showed high levels of resistance to FHB.Lines TA3499 and TA3695 were shown to contain one pair of wheat-Th. elongatum translocated chromosomes involving fragments of 7D plus a segment of the 7E,while line TA3493 was found to contain one pair of wheat-Th.elongatum translocated chromosomes involving the D- and A-genome chromosomes of wheat.Thus,this study has established that the short-arm of chromosome 7E of Th.elongatum harbors gene(s) highly resistant to the spreading of FHB,and chromatin of 7E introgressed into wheat chromosomes largely retained the resistance,implicating the feasibility of using these lines as novel material for breeding FHB-resistant wheat cultivars.     

Identification, mapping, and application of polymorphic DNA associated with resistance gene Pm21 of wheat.

A new powdery mildew resistance gene designated Pm21, from Haynaldia villosa, a relative of wheat, has been identified and incorporated into wheat through an alien translocation line. Cytogenetic and biochemical analyses showed that chromosome arms 6VS and 6AL were involved in this translocation. Random amplified polymorphic DNA (RAPD) analysis was performed on recipient wheat cultivar Yangmai 5, the translocation line, and H. villosa with 180 random primers. Eight of the 180 primers amplified polymorphic DNA in the translocation line, and the same results were obtained in four replications. Furthermore, RAPD analysis was reported for substitution line 6V, seven addition lines (1V-7V), and the F1, as well as F2 plants of (translocation line x 'Yangmai 5'), using two of the eight random primers. One RAPD marker, specific to chromosome arm 6VS, OPH17-1900, could be used as a molecular marker for the detection of gene Pm21 in breeding materials with powdery mildew resistance introduced from H. villosa. Key words : RAPD analysis, 6VS-specific marker, Pm21, Erysiphe graminis f.sp. tritici, Triticum aestivum - Haynaldia villosa translocation.     

三个小黑麦花粉株系的染色体组成分析与抗白粉病鉴定

对来自小黑麦与小麦杂种的３个花粉株系,ＤＨ２２０－４,ＤＨ２２０－５和ＤＨ２２０－１４进行了形态性状观察,染色体组成分析和抗白粉病鉴定。经过染色体形态和数目观察、原位杂交、Ｃ－分带、同工酶等电聚焦和贮藏蛋白的ＳＤＳ－聚丙烯酰胺凝胶电泳分析,证明其中两个株系,ＤＨ２２０－４和ＤＨ２２０－５是６Ｒ／６Ｄ代换系,另一个株系ＤＨ２２０－１４是１Ｒ／１Ｄ代换系。经人工接种鉴定,两个６Ｒ／６Ｄ代换系高抗白粉病。从而进一步证明黑麦的６Ｒ染色体上存在抗白粉病的基因。同时还对小麦遗传背景下异源染色体的识别及６Ｒ染色体的利用价值等问题进行了讨论。