六倍体山羊草与普通小麦杂种F1细胞学研究

粗厚山羊草［Ａｅｇｉｌｏｐｓｃｒａｓｓａ Ｂｏｉｓｓ．（２ｎ＝ ６ｘ＝ ４２,ＤＤＤ２ Ｄ２ＭＣｒＭＣｒ）］、叙利亚山羊草［Ａｅ．ｖａｖｉｌｏｖｉｉ （Ｚｈｕｋ．）Ｃｈｅｎｎ．（２ｎ＝ ６ｘ＝ ４２, ＤＤ ＭＣｒＭＣｒＳＰ ＳＰ）］与普通小麦［Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．（２ｎ＝ ６ｘ＝ ４２, ＡＡＢＢＤＤ）］杂种Ｆ１ 植株形态大多偏向山羊草亲本。４ 个叙利亚山羊草×普通小麦和１ 个粗厚山羊草×普通小麦杂种Ｆ１ 自交结实,结实率在０．１％ —６．５％ 之间。这些种子胚乳很少,生活力较弱,播种后,只有少数种子出苗。杂种Ｆ１ ＰＭＣ减数分裂染色体配对水平较低,出现大量的单价体,二价体低于理论值,并且大多为棒形,说明两种山羊草的Ｄ组染色体已经过很大的修饰。在各杂种Ｆ１ 中还观察到少量的三价体,有些杂种还可见频率很低的四价体和五价体。以山羊草为母本的杂种Ｆ１ 染色体交叉频率优于反交。Ｆ１ 染色体分离极不正常,产生大量的多分孢子和微核。在叙利亚山羊草×冀麦３０ 号中还发现１ 株体细胞染色体为２１ 条的植株,其原因尚需进一步研究确定     

普通小麦三个基因组之间的遗传关系及原位杂交分析

以普通小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍＬ．）的３个可能的二倍体供体种（乌拉尔图小麦（Ｔ．ｕｒａｒｔｕＴｈｕｍ．）拟斯卑尔脱山羊草（ＡｅｇｉｌｏｐｓｓｐｅｌｔｏｉｄｅｓＴａｕｓｃｈ）和粗山羊草（Ａｅ．ｔａｕｓｃｈｉｉＣｏｓｓ．）的基因组ＤＮＡ为研究对象,通过它们之间的相互杂交,比较杂交强度以及泳道中带纹的不同,并结合部分ＤＮＡ重复序列在基因组间含量差异的数据,得出结论：Ａ＾ｕ和Ｄ基因组的关系     

异源细胞质小麦耐盐性的研究

以粗厚山羊草（Ａｅｇｉｌｏｐｓ ｃｒａｓｓａ）细胞质小麦为材料,采用组织培养,水培、模拟盐池等方法,研究细胞质对小麦耐盐性的遗传效应,结果表明：粗厚山羊草细胞质可以不同程度地诱发小麦耐盐性产生变异。不同核质组合细胞质效应有一定差异,表现出特定的核质互作关系。部分异质系细胞水平与植株水平的耐盐性表现一致,特别是（Ａｅ．ｃｒａｓｓａ）－鉴２６的愈伤组织和幼苗的耐盐性表现均最突出。返青期和成熟期的鉴定结     

普通小麦基因组最可能的4个供体的ITS1和ITS2序列及其亲缘关系

对普通小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）基因组（ＡＡＢＢＤＤ）最可能的供体－Ｔ．ｕｒａｔｒｔｕＴｈｕｍ．（ＡＡ）、Ｔ．ｍｏｎｏｃｃｕｍｖａｒ．ｂｏｅｏｔｉｃｕｍ（Ｂｏｉｓｓ．）ＭＫ（ＡＡ）、ＡｅｇｉｌｏｐｓｓｐｅｌｔｏｉｄｅｓＴａｕｓｃｈ．和Ａｅ．ｔａｕｓｃｈｉｉ（Ｃｏｓｓ．（ＤＤ）的核糖体ＲＮＡ基因ＩＴＳ区进行了ＰＣＲ扩增和克隆,并测定了ＩＴＳ１和ＩＴＳ２的ＤＮＡ序列,讨论和纠正了前人     

山羊草属五个基本基因组系统发育的RAPD分析

利用ＲＡＰＤ技术,从ＯＰＥ、ＯＰＦ、ＯＰＧ、ＯＰＵ、ＯＰＸ、ＯＰＹ和ＯＰＺ共７组随机引物中筛选出２８个能产生基因组特异带的稳定引物,对山羊草属（ＡｅｇｉｌｏｐｓＬ．）的５个基本基因组及普通小麦“中国春”的ＤＮＡ进行随机扩增,根据扩增的４８８条ＤＮＡ片段绘制出系统发育图。普通小麦ＡＢＤ基因组与Ｓ基因组亲缘关系最近,Ｃ与Ｕ基因组具有比较近的亲缘关系,Ｄ基因组与其它基因组的亲缘关系比较远     

普通小麦与粗山羊草属间杂种的染色体构型及其后代的育性特征

利用３个推广品种（莱州９５３、山农辐６３、陕７８５９）分别与原产地不同的抗白粉病的６份粗山羊草［Ａｅｇｉｌｏｐｓｔａｕｓｃｈｉｉ（Ｃｏｓｓ．）Ｓｃｈｍａｌ．］杂交,得到６３个无胚乳的种子,将５６枚幼胚接种到Ｎ６＋０．５ｍｇ／ＬＩＢＡ＋０．２ｍｇ／ＬＮＡＡ的培养基上进行褓姆培养,得到３７个植株。其中莱州９５３与粗山羊草的杂交结实率和成苗率较高,分别平均为８．５８％和４．８２％。粗山羊草对白粉病的抗性     

D^2型小麦细胞质雄性不育系及其保持系和恢复系线粒体DNA的比较研究

ｍｓＤ２－ＣＡ８０５７是新育成的具有粗厚山羊草（Ａｅ．ｃｒａｓａ,６ｘ）胞质的Ｄ２型小麦细胞质雄性不育系。采用ＲＦＬＰ和ＲＡＰＤ方法对该不育系及其具有普通小麦（Ｔ．ａｅｓｔｉｖｕｍ）胞质的保持系ＣＡ８０５７和恢复系保－７６９－２２－６的线粒体ＤＮＡ进行分析和比较,发现该不育系的线粒体ＤＮＡ组织结构明显不同于其保持系,也不同于其恢复系。Ｓｏｕｔｈｅｒｎ结果表明,该不育系线粒体基因组在ａｔｐＡ、ａｔｐ９、ｃｏｂ和ｃｏｘⅡ基因上或附近具有显著的组织结构差异。ＲＡＰＤ分析证实了这一点。相反,ＲＦＬＰ和ＲＡＰＤ结果都表明保持系与恢复系之间线粒体基因组结构非常相似。这支持了该不育系的胞质遗传特点来源于与普通小麦胞质差异较大的野生型胞质的事实。推测这种胞质差异与育性有关     

异细胞质信息八倍体小偃麦（Trititrigia 8x）的选育及其性状与细胞遗传

本实验用普通小麦“中国春”和八种异细胞质“中国春”（Ａｅｇｉｌｏｐｓ ｖａｖｉｌｏｖｉｉ）ＣＳ,（Ａｅ．ｊｕｖｅｎａｌｉｓ）ＣＳ,（Ａｅ．ｃｒａｓｓａ）ＣＳ,（Ａｅ．ｃｏｍｏｓａ）ＣＳ,（Ａｅ．ｕｎｉａｒｉｓｔａｔａ）ＣＳ,（Ａｅ．ｓｐｅｌｔｏｉｄｅｓ．Ｍ．）ＣＳ,（Ａｅ．ｋｏｔｓｃｈｙｉ）ＣＳ．（Ｔ．ｔｉｍｏｐｈｅｅｖｉ）ＣＳ分别与八倍体小偃麦（Ｔｒｉｔｉｔｒｉｇｉａ ８ｘ）“远中２”、“远中４     

Bt叶绿体转基因植株的抗虫性及后代表型分析

将Ｂｔ ＣｒｙＩＡ（ｃ） 基因与水稻（ Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．） 叶绿体ｐｓｂＡ 基因的启动子和终止子构建成表达盒,连同烟草（ Ｎｉｃｏｔｉａｎａｔａｂａｃｕｍ Ｌ．） 叶绿体基因组同源片段ｒｐｌ２＿ｔｒｎＨ＿ｐｓｂＡ和ｔｒｎＫ＿ＯＲＦ５０９Ａ 以及选择标记基因ａａｄＡ一起构建成烟草叶绿体转化载体ｐＴＲＳ８。基因枪法转化烟草叶片,经壮观霉素筛选获得转化再生植株。有些转基因植株对３龄棉铃虫（ Ｈｅｌｉｃｏｖｅｒｐａ ｚｅａ） 具有较强的毒杀作用,并能显著抑制昆虫蜕皮和生长发育。对高抗虫性植株的子一代（Ｔ１） 和子二代（Ｔ２） 进行的遗传学和分子生物学分析表明,Ｂｔ 基因已稳定地遗传给子代叶绿体,且抗生素抗性遗传遵循非孟德尔的母系遗传规律     

导入小麦的外源染色体片段的准确鉴定及外源抗性基因的稳定性分析

用抗白粉病的普通小麦一簇毛麦６ＶＳ／６ＡＬ易位系与普通小麦品种扬麦５号、普通小麦一簇毛麦６Ｖ代换系和中国春６Ａ双端二体以及６Ｖ代换系和６Ａ双端二体配制了４个测交组合,分析了这４个杂交组合Ｆ１ＰＭＣ’ｓＭＩＣ－分带的减数分裂构型。在（６ＶＳ／６ＡＬ易位系×扬麦５号）和（６ＶＳ／６ＡＬ易位系×６Ｖ代换系）的Ｆ１ＰＭＣ’ｓＭＩ,分别观察到由易位染色体与６Ａ染色体和６Ｖ染色体配对形成的具有特定Ｃ－分带带型的棒状二价体,杂种中的棒状二价体数目高于各自亲本中的棒状二价体数。在（易位系×Ｃ．Ｓ．ｄ．ｄ．ｔ６Ａ）Ｆ１中,在８７．９％的ＰＭＣ中观察到由易位染色体长臂与６ＡＬ端体配对形成的异形二价体（ｔｌ”）。而在（６Ｖ代换系×Ｃ．Ｓ．ｄ．ｄ．ｔ６Ａ）Ｆ１中,９６．６８％的ＰＭＣ具有两个单价端体（ｔ’,ｔ’）。该结果进一步证实易位涉及簇毛麦染色体６ＶＳ和小麦染色体６ＡＬ,易位断点靠近着丝粒。在减数分裂中,易位染色体的正常配对和分离,保证了６ＶＳ上白粉病抗性基因Ｐｍ２１的正常传递,为这一新抗源在小麦育种中的应用奠定了细胞学基础。     

Agronomic Characteristics and Cytogenetic Observation of the Hybrid F1 from Triticum aestivum and Aegilops triuncialis

In order to efficiently introduce the genes of Aegilops triuncialisL. for resistance to powdery mildew into Triticum aestivum L., it is of importance to understand the genetic mechanism of their F 1 hybrid. It was shown that the bivalent frequency was higher than that of the theoretical value. It resulted from the combination of the wheat inhibitors of 5B Ph gene which located respectively on C and U genome of Aegilops triuncialis L. The results of chromosome in situ hybridization with the C genome-specific repetitive sequence, pAeca212, as the probe further indicated that some chromosomes of the C genome of Ae. caudata L. paired with the chromosomes of the other genomes.     

Multiple origins of allopolyploid <Emphasis Type="Italic">Aegilops triuncialis</Emphasis>

Polyploidization is a key component of plant evolution. The number of independent origins of polyploid species traditionally has been underestimated. The objective of this study was to ascertain the number of origins of a tetraploid Aegilops species. We screened 84 primer sets to identify genome-specific primer sets for the tetraploid wheat relative [Aegilops triuncialis (UUCC genome)] and its diploid progenitors [Ae. umbellulata (UU genome) and Ae. caudata (CC genome)]. Primer sets G12 and G43 were U genome-specific and D21 was a C genome-specific primer. DNA sequence comparison of the G43 locus was used to estimate the number of polyploidization events in the formation of Ae. triuncialis. Parsimony analysis of G43 data revealed at least two independent formations of Ae. triuncialis. In the chloroplast hotspot region, located between genes rbcL and petA, sequence analysis suggested that at least three polyploidization origins might have occurred independently. Ae. triuncialis appears to be a tetraploid derived from multiple origins with minimal genome change after its formation.     

杀配子染色体特异RAPD 标记及山羊草属与普通小麦间分子多态性的研究

以普通小麦"中国春"、三个"中国春"具杀配子染色体的二体异附加系及作为三个杀配子基因种源的三种山羊草为材料, 用46 个10 nt 随机引物对基因组DNA 进行扩增, 以筛选杀配子染色体特有的RAPD 标记, 并检测普通小麦与三种山羊草之间的RAPD 多态性。结果表明:46 个引物中有35 个扩增出比较稳定的RAPD 产物。其中引物OPF-14 和OPQ-09 分别在普通小麦"中国春"-山羊草附加系间扩增出多态性产物;因而认定OPF-14₁₃₀₀ 与OPQ-09₈₀₀、OPF-14₁₁₆₀ 与OPQ-09₇₇₀、OPF-14₁₂₈₀分别为三个杀配子染色体3C、Gcl 和2C 的特异性RAPD 标记, 可以用于快速跟踪鉴定3C、Gcl、2C 杀配子染色体。对三种山羊草与普通小麦"中国春"进行了基因组DNA多态性分析, 结果表明, 35 个引物在"中国春"中共扩增出162 个产物, 在离果山羊草、拟斯卑尔脱山羊草、柱穗山羊草中分别扩增出140、154 和155 个产物;三种山羊草与"中国春"之间的共有扩增产物分别为69、87、96 个, 占总扩增产物的29.61%、37.70%和43.44%。上述结果, 从分子水平揭示了山羊草属与普通小麦之间存在着较近的亲缘关系。     

山羊草属核型分析及其与小麦属的进化关系

作者研究了山羊草属(Aegilops)中的新疆节节麦(Ae.squarrosa)、拟斯卑尔脱山羊草(Ae.speltoides)、沙融山羊草(Ae.sharonensis)、尾状山羊草(Ae.caudata)、卵圆山羊草(Ae.ovata)、偏凸山车草(Ae.ventricosa),钩状山羊草(Ae.triuncialis)、三芒山羊草(Me.triaristata)、欧山羊草(Ae.biuncialis)、柱穗山羊草(Ae.cylindrica)、可兹山羊草(Ae.kotschyi)和肥厚山羊草(Ae.crassa)的核型和部分材料的Giemsa N-带,结果表明山羊草属的C组核型为:4sm+3st;D组核型为:6m+1sm;S组的核型为:6m+1sm;M组的核型为:4m+1sm+2t。在四倍体、六倍体中,各染色体组保持着相对稳定。山羊草属S、D染色体组的核型与带型表明它们是小麦B、D染色体组的可能供体,C、M染色体组的一部分染色体带型亦与小麦B组带型相似。     

Molecular-genetic analysis of wheat (T. aestivum L.) genome with introgression of Ae. cylindrica Host genetic elements

Wheat-aegilops hybrid plants Triticum aestivum L. (2n = 42) x Aegilops cylindrica Host (2n = 28) were investigated with using microsatellite markers. In two BC1F9 lines some genome modifications connected with losing DNA fragments of initial variety or appearing of Aegilops genome elements were detected. In some investigated hybrids new amplicons lacking in parental plants were found. Substitution of wheat chromosomes for aegilops chromosomes was not revealed. Analysis of microsatellite loci in BC2F5 plants showed stable introgression of aegilops genetic elements into wheat; elimination of some transferred aegilops DNA fragments in the course of backcrossing; decreasing size of introgressive elements after backcrossing. Introgressive lines were classified according to genome changes.     

AFLP-based analysis to study genetic variability and relationships in the Spanish species of the genus Aegilops

Amplified fragment length polymorphism (AFLP) DNA markers were used to characterize the genetic diversity and relationships in wild species of the genus Aegilops. Fifty populations, which included the species Aegilops biuncialis (UUMM), Ae. neglecta (UUMMNN), Ae. ovata (UUMM), Ae. ventricosa (DDNN) and Ae. triuncialis (UUCC) were selected. These populations are distributed in the Iberian peninsula and Balearic islands. Five AFLP selective primer combinations generated a total of 527 amplification products of which 517 (98.10%) detected polymorphisms. Aegilops neglecta showed the least variation in contrast with Ae. biuncialis that presented the highest degree of polymorphism. Genetic relationships within the populations were evaluated by generating a similarity matrix based on the Jaccard index. In the resulting phenogram Ae. ventricosa appears segregated from the other species, probably owing to the influence of the D genome. The species sharing the U genome are located in the main cluster. The branching pattern of the U genome group reflects the proximity of the species sharing the M genome. Ae. biuncialis and Ae. ovata are clearly separated suggesting that the super index system should be used to differentiate the M genomes of both species. The variation among populations within species in relation to their geographical origin and results previously obtained by the authors using biochemical and molecular markers are discussed.     

Chromosome pairing in the allotetraploid Aegilops biuncialis and a triploid intergeneric hybrid.

Chromosome pairing behaviour of the natural allotetraploid Aegilops biuncialis (genome UUMM) and a triploid hybrid Ae. biuncialis x Secale cereale (genome UMR) was analyzed by electron microscopy in surface-spread prophase I nuclei. Synaptonemal-complex analysis at zygotene and pachytene revealed that synapsis in the allotetraploid was mostly between homologous chromosomes, although a few quadrivalents were also formed. Only homologous bivalents were observed at metaphase I. In contrast, homoeologous and heterologous chromosome associations were common at prophase I and metaphase I of the triploid hybrid. It is concluded that the mechanism controlling bivalent formation in Ae. biuncialis acts mainly at zygotene by restricting pairing to homologous chromosomes, but also acts at pachytene by preventing chiasma formation in the homoeologous associations. In the hybrid the mechanism fails at both stages. Key words : Aegilops biuncialis, allotetraploid, intergeneric hybrid, pairing control, synaptonemal complex.     

Heat shock of cultivated and wild wheats during early seedling stage: growth, cell viability and heat shock proteins

Growth, cell viability and heat shock proteins (HSPs) in Bezostaya-1, Cukurova-86 and Diyarbakir-81 cultivated wheat cultivars and three Aegilops species were investigated. Etiolated seedlings were exposed to 23 degrees C, 32 degrees C, 35 degrees C, 37 degrees C and 38 degrees C for 24 h, and 35 degrees C (24 h) --> 50 degrees C (1 h) and 37 degrees C (24 h) --> (50 degrees C (1 h). At the end of recovery growth periods, the shoot lengths of the genotypes generally decreased significantly at 35, 37 and 38 degrees C. The acquired thermal tolerance (ATT) in intact seedlings was over 50% at 35 degrees C --> 50 degrees C and 37 degrees C --> 50 degrees C, but in cell viability test it ranged from 2.75% (Ae. triuncialis) to 32.87 (Bezostaya-1) at 35 degrees C, and from 2.82% (Ae. triuncialis) to 37.82 (Bezostaya-1) at 37 degrees C. Ae. triuncialis was most sensitive genotype in both ATT determination. In electrophoretic profiles of proteins, while some HSPs were newly synthesized, some normal cellular proteins disappeared at 37 degrees C and 37 degrees C -->50 degrees C compared to 23 degrees C. The number of low molecular weight (LMW) HSPs were more than intermediate- (IMW) and high- (HMW) HSPs. The genotypes had both common (12 HSPs between at least two genotypes) and genotype-specific (33 HSPs) LMW HSPs. The common HSP of 19.8 kDa (pI 6.5) was synthesized in Bezostaya-1, Cukurova-86, Diyarbakir-81, Ae. biuncialis and Ae. umbellulata. Bezostaya-1 is the only genotype that synthesized 12 IMW and 2 HMW HSPs at 37 degrees C --> 50 degrees C. Ae. triuncialis had only two common LMW HSPs [22.1 (pI 7.1) and 24.2 kDa (pI 6.5)].     

Multiple origins promote the ecological amplitude of allopolyploid Aegilops (Poaceae)

Polyploidy has been ubiquitous in plant evolution and is thought to be an important engine of biodiversity that facilitates speciation, adaptation, and range expansion. Polyploid species can exhibit higher ecological tolerance than their progenitor species. For allotetraploid species, this higher tolerance is often attributed to the existence of heterosis resulting from entire genome duplication. However, multiple origins of allopolyploid species may further promote their ecological success by providing genetic variability in ecological traits underlying local adaptation and range expansion. Here we show in a group of allopolyploid species in the genus Aegilops that range size and abundance are correlated with the number of inferred origins. We found that allopolyploid Aegilops spp. contain multiple chloroplast haplotypes, each identical to haplotypes of the diploid progenitor species, indicating multiple origins as the major source of variation. The number of inferred origins in each allopolyploid species was correlated to the total area occupied by the allopolyploid and the tendency for the species to be common. Additionally, we found differences in ecological tolerance among independent origins in Aegilops triuncialis. These results strongly support the hypothesis that the introduction of genetic variability by multiple origins can increase the ecological amplitude and evolutionary success of allopolyploid species.     

Genome differentiation in Aegilops. 1. Distribution of highly repetitive DNA sequences on chromosomes of diploid species.

Genome differentiation in 12 diploid Aegilops species was analyzed using in situ hybridization with the highly repetitive DNA sequences pSc119 and pAs1 and C-banding. Chromosomes of all these diploid Aegilops species hybridized with the pSc119 probe; however, the level of hybridization and labeling patterns differed among genomes. Only four species (Ae. squarrosa, Ae. comosa, Ae. heldreichii, and Ae. uniaristata) showed distinct hybridization with pAs1. The labeling patterns were species-specific and chromosome-specific. Differences in in situ hybridization (ISH) patterns, also observed by C-banding, exist between the karyotypes of Ae. comosa and Ae. heldreichii, suggesting that they are separate, although closely related, subspecies. The S genome of Ae. spelioides was most similar to the B and G genomes of polyploid wheats on the basis of both C-banding and ISH patterns, but was different from other species of section Sitopsis. These species had different C-banding patterns but they were similar to each other and to Ae. mutica in the distribution of pSc119 hybridization sites. Two types of labeling were detected in Ae. squarrosa with the pAs1 probe. The first resembled that of the D-genome chromosomes of bread wheat, Triticum aestivum L. em. Thell., while the second was similar to the D genome of some of the polyploid Aegilops species. Relationships among diploid Aegilops species and the possible mechanisms of genome differentiation are discussed. Key words : wheat, Triticum, Aegilops, in situ hybridization, C-banding, evolution.