小麦-黑麦小片段易位系新种质的分子细胞遗传学鉴定

为创制有利用价值的小麦-黑麦易位系,选育小麦遗传改良的新种质,本研究运用形态学与分子细胞遗传学结合的研究手段,对小麦-黑麦代换系6R/6A×克珍杂种后代中选育的11个品系进行鉴定。结果表明,11个品系形态表现为大穗多花、多小穗、抗倒伏、籽粒饱满度好等优良性状。其中有4个品系2-13、2-18、2-20、2-21穗长超过14 cm。品系2-18、2-21,分蘖数为7个,2-17千粒重为35.24 g,品系2-19主穗粒数为54,均超过亲本。另有8个品系结实率也优于亲本。亲本克珍为小麦-黑麦小片段易位系,带有7R和3R染色体遗传成分,分子标记和原位杂交显示,11个品系均带有R组染色体成分,分别有10个、6个和3个品系带有黑麦6R、7R和3R染色体遗传成分。有9个品系可确定为小麦-黑麦小片段易位系,易位类型为端部易位和中间易位。本研究成果对小麦育种及种质改良具有重要意义。     

小麦EST-SSR标记的开发和染色体定位及其在追踪黑麦染色体中的应用

根据小麦盐胁迫诱导和茎秆组织相关EST序列开发了81对EST-SSR引物, 其中67、46、18和61对分别在小麦、黑麦、簇毛麦和大麦基因组中稳定扩增, 在不同小麦和大麦品种间具有多态性的引物分别有22和23对。利用小麦缺体-四体系共定位了43对引物的81个位点, 其中A、B和D染色体组上分别有29、30和22个位点, 涉及除4B、3D和6D外的18条染色体。此外30对引物在黑麦基因组中具有特异扩增, 其中8对分别在黑麦1R、4R、5R和R7染色体上具有特异扩增, 7对在多条黑麦染色体具有相同扩增。这些新标记可有效用于小麦及其近缘物种的遗传作图与比较遗传研究。     

异源转座子存在的证据

本研究选用130对小麦SSR引物对从5个小麦-黑麦易位系中选取的56个单株进行PCR扩增,中国春,绵阳11和黑麦作为对照。结果发现122-2A和636-2A引物对在小麦和黑麦间产生多态性。122-2A引物在黑麦中产生2个黑麦特异条带,一个条带大小为190bp,另一个为330bp。在56个单株中有4个单株产生相应的黑麦特异带。这可能是黑麦中1个可移动因子(异源转座子)导入了小麦所致。并且结合许多病理学、进化生物学及其遗传学的证据对异源转座子在原核和真核生物中广泛存在的可能性进行了讨论。     

小麦-黑麦二体代换系间杂交诱导染色体易位的研究

摘 要：小麦-黑麦二体代换系间杂交,是诱导小麦-黑麦染色体易位的重要途径,有理论与应用价值。本研究利用DS1R/1D与DS5R/5A、DS6R/6A杂交,结果表明,F1减数分裂有19个二价体、4个单价体,有部分同源染色体配对和染色体易位现象;F2 对带有黑麦性状的24株普通小麦类型植株进行原位杂交检测,共检测出10株有染色体易位,易位频率为41.6%。易位株系为：06-16-7、06-16-3、06-16-5、06-16-7、06-16-8、06-17-7、0617-11、06-17-15、06-17-16、06-18-5。由于亲本选配、检测植株有一定的目的性,易位植株均表现有一定的应用价值。     

杀配子染色体在小麦-黑麦异代换系中的作用研究

利用小麦-黑麦异代换系H-13(1R/1D), H-24(5R/5A),H-33(6R/6A)与中国春-杀配子染色体二体异附加系CS-DA2C(来自山羊草Ae. cylindrica),CS-DA3C(来自山羊草Ae. triuncialis)配置杂交组合,对杀配子染色体的作用进行了研究。结果表明,杀配子染色体3C对H-13和H-24都是致死的,即有着完全的杀配子作用,但对H-33的杀配子作用则是不完全的,有着接近正常的结实率,说明不同的小麦-黑麦异代换系遗传背景对杀配子作用有很大影响,可能在品系H-33中存在对2C染色体杀配子作用的抑制基因。在以品系H-33为母本时,2个杀配子附加系F1的自交结实率差异显著,说明在相同的小麦-黑麦异代换系遗传背景下,染色体2C与3C的杀配子作用是不同的,在这2个杀配子染色体上可能带有不同的杀配子基因?这些研究结果为进一步创制小麦-黑麦易位系奠定了基础。     

黑麦重复序列在检测小麦品种中外源染色体的应用

本研究根据RAPD引物OPH20在黑麦中扩增出的特异序列pSc20H.2设计一对PCR引物pSc20ht-23/24,以来源于黑麦的小麦抗叶锈近等基因系材料TcLr45及感病对照Thatcher为亲本进行PCR扩增。并对42个小麦抗叶锈近等基因系及103个小麦品种材料进行检测。引物pSc20ht23/24在TcLr45中扩增出一条约750bp的条带,而在Thatcher中无扩增条带。对该特异片段回收、克隆测序为734bp。42个小麦抗叶锈近等基因系检测在TcLr26中扩增出与TcLr45相同的条带,而在同样来源于黑麦的小麦抗叶锈近等基因系TcLr25中未扩增出该条带;中国春-Imperial黑麦附加系1R-7R中除5R外均扩增出该条带;13个1B/1R易位系小麦品种也扩增出该条带;90个地方小麦品种中有16个扩增出该条带,6个品种经系谱分析具有黑麦遗传背景,表明该标记可用于检测小麦中含有的除黑麦5R染色体之外的外源染色质。     

燕大1817/北农6号重组自交系群体穗部性状的QTL定位

小麦穗部性状与单株产量密切相关。本研究以小麦骨干亲本燕大1817与优良品系北农6号衍生的269个重组自交系为材料,通过在北京和河北石家庄的2年田间试验数据,利用本实验室已构建的高密度SNP和SSR遗传连锁图谱进行穗长、穗粒数和穗粒重QTL定位。采用完备复合区间作图法共检测到29个穗部性状加性效应QTL,其中10个穗长QTL分布于1B、2D、3A、3B、4A、5A、5B、6A和7D染色体上,解释的表型变异率为2.96%~9.63%,QSl.cau-4A.2在所有5个环境中均能被检测到,解释的表型变异为5.89%~9.62%,另有7个QTL能在2个或2个以上环境中被检测到;8个穗粒数相关QTL分布于1A、3A、3D、4A和5B染色体上,解释的表型变异为4.06%~11.17%,为单个环境QTL。11个与穗粒重相关QTL分布于1A、1B、2A、2D、3A、4D、5A、5B和6B染色体上,解释的表型变异为2.79%~16.12%,其中QGws.cau-1B、QGws.cau-3A和QGws.cau-6B.2在2个或者2个以上环境中能被检测到。另外,鉴定出6个分布于1A、2D、3A、4A和5B染色体上的QTL富集区段。     

小麦品种百农AK58与周麦18的遗传差异性分析

为解析百农AK58和周麦18的遗传差异,利用表型数据分布于小麦全基因组的305个SSR标记进行分析,以期为小麦大面积种植品种的培育提供参考。方差分析表明,百农AK58和周麦18在株高、千粒质量和单位面积穗数上存在极显著差异,其中百农AK58在株高和单位面积穗数上明显优于周麦18。分子标记比较发现,305对SSR引物中有130对引物在百农AK58与周麦18之间扩增出差异带,多态引物比例达到42.6%。2个品种在A、B、D基因组存在差异的引物数基本一致,分别有34,37,37个,多态引物比例表现为B基因组(43.9%)A基因组(37.1%)D基因组(33.5%)。百农AK58和周麦18的遗传差异在小麦7个部分同源群和21对染色体上也表现一定的不均衡性。在7个部分同源群之间,二者在第5部分同源群的遗传差异最大,多态引物比例达到49.3%,而在染色体水平上,它们在3B、5B、7B、4A的遗传差异相对较大,多态性引物比例均超过50%。百农AK58和周麦18所不同的SSR位点是遗传研究中需要重点关注的基因组区域。     

太谷核不育小麦衍生材料川6415在其后代中的遗传分析

川6415是利用太谷核不育小麦育成的重要小麦育种基因资源,解析川6415在其衍生后代的遗传,对利用川6415进行分子标记辅助选择育种具有重要意义.本研究选用617对SSR引物对川6415及其衍生品种(系)进行扫描,分析川6415在其一代、二代衍生品种(系)的遗传,结果表明,一代衍生品种川麦42继承了26.6％川6415的遗传物质;二代衍生品种(系)31区和R104更多的继承了川6415的遗传物质,分别为32.6％和37.2％.川麦42遗传背景中来源于川6415的SSR标记位点,在除3D、4D、7D外的18条染色体上都有分布,在2A、2B和4B染色体上形成染色体区段.4B上川麦42遗传背景中源于川6415的染色体区段有利于增加穗数/m2; 2B和7A上的川6415染色体区段有利于降低川麦42株高.因此,太谷核不育小麦衍生材料川6415可作为增加穗数/m2和降低株高的重要基因资源用于小麦高产育种.     

山农01-35×藁城9411重组自交系遗传图谱构建及粒重QTL分析

为利用分子遗传图谱进行小麦产量数量性状位点定位分析,以大粒高产小麦品系山农01-35和强筋小麦藁城9411为亲本,衍生了含182个家系的重组自交系(RIL)F₈群体,用442个DArT标记、59个SSR标记和1个TaGW2-CAPS标记,构建了包括29个连锁群的分子遗传图谱,总遗传长度为4 084.5 cM,标记间平均距离为8.13 cM。定位了54个新标记位点,包括44个DArT和10个SSR标记,分布于除4D、6B、7B外的其他18条染色体上。用该分子遗传图谱和4个环境粒重表型值,共检测到7个影响粒重的加性QTL,分别位于1B、4B、5B、6A染色体,其中QGW4B-7、QGW5B-20和QGW6A-29在单环境分别定位和4个环境共同定位两种方法中均能检测到。QGW4B-7、QGW5B-12和QGW6A-29对粒重的贡献率均超过10%,为主效QTL。本研究结果可为小麦高产优质性状的QTL分析及分子标记辅助选择提供参考。     

Chromosomal location of factors affecting content and composition of non-starch polysaccharides in wheat-rye addition lines

Summary Content of dietary fibre (DF) as well as content and composition of non-starch polysaccharides (NSP) were evaluated in disomic wheat-rye addition lines and octoploid triticale compared to their parental species — wheat (Grana) and rye (Dakowskie Zote). Large variation in the contents of NSP and DF were observed in the wheat-rye addition lines, especially in the soluble fractions. The double addition of rye chromosomes led to important transgression effects on content and composition of DF in wheat grain. Chromosomes 2R and 5R increased SDF content above the rye level, whereas the 3RS arm decreased SDF content below the wheat level. The octoploid triticale demonstrated the highest content of total arabinoxylan (AX), exceeding that of rye, while the double addition of chromosomes 4R, 6R and 6RL had an impact on high expression, comparable to that of rye content of total AX in wheat grain. Chromosomes 2R and 5R notably increased the proportion of soluble non-cellulosic glucose in the NSP fraction, in contrast to the rest of wheat-rye addition lines, octoploid triticale, wheat and rye, where AX was found to be predominant among NSP constituents. The effects of single chromosome pairs on content and composition of NSP proved to be higher than the effect of the whole rye genome in octoploid triticale. The most remarkable effect, especially considering the direction of changes, was of a 3R chromosome short arm addition. The obtained data might be of interest in breeding rye or triticale with lower viscous AX content as well as rich in soluble non-starch glucose polymers, regarded as a corrective factor in modern diseases.Abbreviations AX arabinoxylan - CV coefficient of variation - DF dietary fibre - IDF insoluble dietary fibre - NSP non-starch polysaccharides - SDF soluble dietary fibre - TDF total dietary fibre - TFA trifluoroacetic acid     

不同供体及不同回交次数对玉米自交系R08的改良效应

以R08为轮回亲本,18个优良自交系为供体亲本,不同回交和自交次数,选育出遗传背景与R08相近、但相互之间又存在一定差异的BC₁F₃和BC₂F₂各18个R08改良系。通过抗病性鉴定、配合力及SSR分子标记分析,探讨不同供体及不同回交次数对R08的改良效果。结果表明,36个改良系中,29个抗或高抗大斑病,大部分改良系的多数产量性状一般配合力(GCA)与R08相比并无下降或有所提高;相同供体不同回交次数选系的比较显示,对大斑病抗性的改良,回交1次自交2次(BC₁F₃)优于回交2次自交1次(BC₂F₂),且改良后代选系多数产量性状GCA大体相当;相同回交次数不同供体选系的比较表明,供体对回交后代的影响较大,供体不同回交后代选系大斑病抗性及多数产量性状GCA存在较大的差异;SSR分子标记研究结果在一定程度上揭示相同供体不同回交次数所创造的遗传变异无明显差异,相同回交次数不同供体选系在分子水平上存在较大差异;供体昌7-2和川321对改良R08的大斑病抗性和产量性状GCA作用较大,属优良供体亲本;w4-1和w10-1属回交改良优良选系。因此,利用回交法改良玉米自交系,在选准供体亲本的基础上,回交1次后,在自交过程中加强目标性状的鉴定选择及配合力测定,可提高回交改良的育种效率。     

Characterization of wheat-triticale doubled haploid lines by cytological and biochemical markers

Five wheat-triticale doubled haploid (DH) lines— M08, V209, DH220-14-2, DH696-3-4 and M16 —derived from anther culture of F₁s resulting from crosses involving hexaploid or octoploid triticale × hexaploid wheat, were characterized by cytological and biochemical markers. Cytological evidence from genomic in situ hybridization and C-banding indicated that DH lines M08 and V209 (2n= 42) each contained a pair of 1BL/1RS translocation chromosomes. DH220-14-2 (2n= 42) was also a translocated line with two pairs of chromosomes containing small fragments of rye. One of the translocation fragments carried the Sec-1R gene originating from the satellite region of 1RS; the origin of the other one remains unknown. DH696-3-4 (2n= 42) contained a 3D(3R) substitution. In M16 (2n= 44), three pairs of rye chromosomes, 3R, 4R and 6R, were present, 4R as an addition and 3D(3R) and 6D(6R) as substitutions. Biochemical, isozyme and storage protein markers confirmed the cytological conclusions. The advantages of transferring alien chromosomes or chromosome fragments into wheat and creating alien aneuploid lines by anther culture of hybrid F¹s are discussed.     

Effect of alien 5R(5A) chromosome substitution on ear-emergence time and winter hardiness in wheat-rye substitution lines

Ear emergence time and response to vernalization were investigated in 12 alien substitution lines in which a pair of chromosomes 5A of recipient spring wheat cultivars was replaced by a pair of chromosomes 5R of Siberian spring rye ‘Onokhoiskaya’. The recipients were 12 spring cultivars of common wheat, each carrying different Vrn genes. Spring rye ‘Onokhoiskaya’ had the Sp1 (now called Vrn-R1) gene for spring growth habit located on chromosome 5R, but its expression was weaker. The Vrn-R1 gene had no effect on growth habit, ear emergence time and response to vernalization in wheat-rye substitution lines. Ears emerged significantly later in the 5R(5A) alien substitution lines than in the recipient wheat cultivars with the Vrn-A1/Vrn-B1/vrn-D1 or Vrn-A1/vrn-B1/Vrn-D1 genotypes. No difference in ear emergence time was found between most of the 5R(5A) alien substitution lines and the cultivars carrying the recessive vrn-A1 gene. The presence of the Vrn2a and Vrn2b alleles at the Vrn2 (now called Vrn-B1) locus located on wheat chromosome 5B was confirmed.The replacement of chromosome 5A by chromosome 5R in wheat cultivars ‘Rang’ and ‘Mironovskaya Krupnozernaya’, which carries the single dominant gene Vrn-A1, converted them to winter growth habit. In field studies near Novosibirsk the winter hardiness of 5R(5A) wheat–rye substitution lines of ‘Rang’ and ‘Mironovskaya Krupnozernaya’ was increased by 20–47% and 27–34%, respectively, over the recurrent parents.     

The effect of single D-genome chromosomes on aluminum tolerance of triticale

Aluminum tolerance of two sets of hexaploidtriticale (×Triticosecale Wittmack)lines with disomic substitutions of theD-genome chromosomes from Triticumaestivum L. was analyzed by themodified-pulse method. Of the 20substitution lines in winter triticalePresto, and 18 lines of spring triticaleRhino, six and nine lines, respectively,showed improved tolerance relative to thatobserved in the control lines. The D-genomechromosomes in substitutions 1D(1B),3D(3A), 3D(3B), 4D(4A), 4D(4B) and 6D(6B)significantly (p<0.01) improved Altolerance in both sets of lines. Highpercentages of tolerant plants were alsoobserved in 2D(2B) and 5D(5A) substitutionsin Presto and in 2D(2A), 2D(2B), 5D(5B),6D(6A) and 7D(7A) substitution lines ofRhino. In no instance, the removal ofindividual rye chromosomes, bysubstitutions, improved Al tolerance of therecipient line. Moreover, the presence of acomplete rye genome, and especially ofchromosome 3R, was necessary for triticale'stolerance to aluminum. The results alsoindicated some effects of allelic variationpresent on both rye and wheat chromosomes,and a possibility of interactions ofvarious factors.     

苦荞全基因组SSR位点特征分析与分子标记开发

目前苦荞SSR多态性标记数量较少,根据已发表的苦荞基因组测序数据,利用MISA软件对1~6核苷酸重复的SSR位点进行了查找和序列特征分析,批量设计引物并对引物进行了有效性和多态性检测。结果表明,苦荞基因组中共检测到1 640个SSR位点,其中三核苷酸重复型SSR最多,占比63.29%,五核苷酸重复型最少,仅占0.12%。AT/TA、AAG/CTT、ACC/GGT和ATC/GAT为出现频率较高的重复基序。苦荞基因组SSR序列长度变化范围为12~476bp,平均长度23.14bp,长度12~19bp的占比71.71%,长度≥20bp的占比28.29%。根据不同类型SSR位点设计并合成引物479对,选择200对引物对5份苦荞资源和3份甜荞资源进行多态性检测,有56对扩增出多态性条带,17对在苦荞种质中产生多态性条带,48对在甜荞种质中产生多态性条带,9对同时在两种种质中产生多态性条带。利用苦荞全基因组序列可实现SSR标记的批量开发,可鉴定出适用于苦荞和甜荞遗传多样性分析、遗传图谱构建和品种鉴定等研究的SSR引物。