应用GISH与STS标记鉴定小麦-中间偃麦草抗黄矮病端体系

由大麦黄矮病毒引起的小麦黄矮病毒病是一个严重病害,至今在小麦属内还没有发现抗源。中间偃麦草2Ai-2染色体携带一个高抗黄矮病基因,对该基因的染色体臂定位将为制定抗病基因向小麦转移策略,筛选、开发特定的、与抗性连锁的分子标记的研究提供重要信息。本文对由小麦-中间偃麦草二体附加系Z6衍生的3个抗黄矮病端体系进行鉴定,通过分析端体的遗传构成、筛选与端体共分离的STS标记以确定端体在遗传上的染色体臂归属,从而明确BYDV抗病基因的染色体位置。以拟鹅冠草基因组[Pseudoroegneria strigosa (M. Bieb.) Löve,St]DNA为探针,中国春基因组(Triticum aestivum L., ABD) DNA作封阻分别对抗病亲本Z6及抗病端体系N530的根尖体细胞染色体进行原位杂交,结果表明,N530体细胞中有2个端体显示出与Z6中外源染色体2Ai-2短臂相似, 而与长臂不同的杂交信号。以小麦第2同源群的5个RFLP探针的DNA序列为基础,设计了6对PCR引物,对小麦-中间偃麦草二体异附加系、二体代换系和端体系进行扩增,结果表明,基于短臂探针psr126,psr131序列设计的2对引物,可在含有2Ai-2染色体及端体的抗黄矮病材料中特异扩增,而基于长臂探针psr112序列设计的1对引物,可在含有2Ai-2染色体的抗黄矮病材料中特异扩增,但不能在端体系进行特异扩增,证明外源端体为2Ai-2染色体的短臂。本研究不仅将黄矮病抗性基因定位于2Ai-2染色体的短臂上, 而且由RFLP探针psr126、psr131和psr112序列转化的标记STS₁₂₆ (sequence tagged site) STS₁₃₁和STS₁₁₂还可分别作为追踪2Ai-2染色体短臂和长臂的分子标记,用于抗病易位系辅助选择。     

用RFLP鉴定普通小麦—纤毛鹅观草二体附加系中外源染色体的同源转化性

用小麦族7个部分同源群的40个RFLP探针对小麦——纤毛鹅观草二体附加系进行分析,在证实了原有细胞学鉴定结果的基础上,又进一步提供了纤毛鹅观草染色体部分同源群的分子证据。即96K025,96K026中添加的一对纤毛鹅观草染色体B属于第2部分同源群;96K012, 96K013中添加的一对染色体E属于第5部分同源群。对以上株系的衍生株系     

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.     

Genetic variation in barley of crossability with wheat and its quantitative trait loci analysis

To study genetic variation in crossability, 80 barley accessions of diverse geographic origin consisting of 50 wild barleys (H. vulgare ssp. spontaneum or ssp. agriocrithon) and 30 cultivated barleys (H. vulgare ssp. vulgare) were crossed as the male parent with a highly crossable wheat variety, Shinchunaga. Crossabilities, expressed as the percentage of pollinated florets giving embryo-containing caryopses, ranged from 0% to 68.6%. Barley accessions from East Asia had generally a low crossability, while barley accessions from other regions exhibited a wider range of crossability including highly crossable genotypes. No significant difference in mean crossability was found between wild and cultivated barleys. To estimate the number and location of barley genes controlling the crossability, doubled haploid lines derived from the cross between the barley varieties Steptoe and Morex were crossed as the male parent with wheat. Quantitative trait loci (QTL) analysis using molecular markers identified four QTL. These were mapped to the centromeric regions of chromosomes 2H, 3H and 5H and the short arm of chromosome 7H. The QTL on chromosomes 3H and 5H had larger effects than those on chromosomes 2H and 7H. The four QTL collectively explained 35.4% of the total variance under a multiple QTL model. Relationships of the QTL identified in the present study with previously reported crossability genes of barley and wheat are discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Development and identification of wheat–Haynaldia villosa T6DL.6VS chromosome translocation lines conferring resistance to powdery mildew

Three lines conferring resistance to powdery mildew, Pm97033, Pm97034 and Pm97035, were developed from the cross of Triticum durum-Haynaldia villosa amphidiploid TH3 and wheat cv.'Wan7107' via backcrosses, immature embryo and anther culture. Genomic in situ hybridization analysis showed that these lines were disomic translocation lines. Cytogenetic analysis indicated that the F1 plants of crosses between the three translocation lines and 'Wan7107' and crosses between the three translocation lines and substitution line 6V(6D) formed 21 bivalents at meiotic metaphase I. Aneuploid analysis with 'Chinese Spring' double ditelocentric stocks indicated that the translocated chromosomes were related to chromosome 6D. Biochemical and restriction fragment-length polymorphism (RFLP) analyses showed that the translocation lines lacked a specific band of 6VL of H. villosa compared with the substitution and addition lines but possessed specific markers on the short arm of the 6V chromosome of H. villosa. The three translocation lines lacked specific biochemical loci and RFLP markers located on chromosome 6DS. The results confirmed that Pm97033, Pm97034 and Pm97035 were T6DL.6VS translocation lines.     

Allocation of a powdery mildew resistance locus to the chromosome arm 6RL of <Emphasis Type="Italic">Secale cereale</Emphasis> L. cv. ‘Jingzhouheimai’

Chinese cultivar Jingzhouheimai of Secale cereale L. confers resistance to several important wheat diseases, making it a useful resource for wheat improvement. Octoploid amphiploid (2n = 8x = 56; AABBDDRR) was synthesized by hybridization of Jingzhouheimai with hexaploid wheat landrace Huixianhong and doubling of the F1 chromosome number. This amphiploid was backcrossed to the wheat parent to produce wheat-rye chromosome addition lines. The existing six disomic addition lines (all except 6R) were screened with 16 rye-specific DNA markers and three putative markers for 6R were identified. These markers were used in selection of chromosome 6R addition lines, confirmed by the genomic in situ hybridization (GISH). In this manner, seven monosomic 6R and five disomic addition were selected, as well as one mono-telosomic 6RS addition, one mono-telosomic and one ditelosomic 6RL additions. In turn these 6R addition lines were used to develop additional 15 6R-specific markers of which six were allocated to the arm 6RS and nine to 6RL. Screening for the powdery mildew resistance identified chromosome arm 6RL as the carrier of the resistance locus. Therefore, DNA markers identified as specific to the 6RL arm can be used to monitor the introgression of the resistance locus into wheat.     

Effects of barley chromosome on heading characters in wheat-barley chromosome addition lines

Heading time in cereals is a composite character determined by vernalization requirement, photoperiodic sensitivity and narrow-sense earliness. To study the effects of added barley chromosomes on the heading characters in wheat, two sets of wheat-barley chromosome addition lines, i.e., ‘Betzes’ barley chromosomes 2H to 7H added to ’Chinese Spring‘ wheat (CS-Be2H to CS-Be7H) and ‘New Golden’ barley chromosomes 5H and 6H added to ‘Shinchunaga’ wheat (Shi-NG5H, Shi-NG6H), were examined for their heading characters. All barley chromosomes except Be6H affected vernalization requirement and/or narrow-sense earliness in CS or Shi. Be5H chromosome also slightly increased the photoperiodic sensitivity of CS. Shi-NG5H addition line showed significantly decreased vernalization requirement in comparison with Shi, whereas CS-Be5H did not show any difference from CS. The F₁ hybrid of the cross, Shi-NG5H × CS-Be5H, exhibited the same level of vernalization insensitivity as the Shi-NG5H addition line, and plants with and without a vernalization requirement segregated in a 1 : 3 ratio in the F₂ generation. These observations, together with previous reports, suggest that the decreased vernalization requirement in the Shi-NG5H addition line was caused by the presence of a major dominant gene for spring habit, Sh₂, located on the NG5H barley chromosome. Furthermore, this study revealed that the Sh₂ gene in barley has a similar but weaker effect than the wheat vernalization insensitive gene, Vrn1, on the vernalization response in wheat. This revised version was published online in July 2006 with corrections to the Cover Date.     

Isolation of a fertile wheat-barley addition line carrying the entire barley chromosome 1H

The isolation of six of the seven possible additions of barley chromosomes to the wheat genome reported 18 years ago has made an important contribution to gene mapping in barley, first with genes controlling isozymes and more recently DNA (molecular) markers. A fertile addition line involving barley chromosome 1H,which carries genes controlling several characters of economic importance, could not be isolated at that time because it caused extreme meiotic abnormalities leading to complete sterility when added to wheat. Later the short arm of barley chromosome 1H was added to wheat as a fertile ditelosomic addition, but the non-availability of the entire barley chromosome 1H addition line has hampered the location of barley genes to the long arm of this chromosome. This problem has now been overcome cytogenetically as described herein. The resultant self-fertile disomic-monotelodisomic addition line carrying a pair of barley chromosome 6H and a heteromorphic 1H/1HS pair is more stable, and makes the wheat-barley addition line series complete for gene mapping work and will provide a vehicle for the possible transfer of useful genes from this barley chromosome to wheat.     

Effect of added barley chromosomes on the flowering time of new wheat/winter barley addition lines in various environments

The heading characters and morphological traits of two partial sets of wheat–barley disomic addition lines, namely Mv9kr1/Igri and Asakaze/Manas, were evaluated under controlled environmental conditions in a phytotron under long-day, short-day and non-vernalised conditions and in field-sown experiments. The winter barley chromosome additions significantly influenced the flowering time of wheat both in the controlled environment test and under field-sown conditions. Of all the barley addition lines, the effect of the 4H and 7H additions was the most characteristic. The 7H addition lines were the earliest in both cultivar combinations in each treatment. In the Mv9kr1/Igri combination the 4H addition was the latest under all the environmental conditions. In the Asakaze/Manas combination 4H addition was the latest under short-day and long-day illumination in the phytotron but the 6H addition was the latest without vernalisation and in the field in 2012. There was 12 and 11 days difference between the flowering times of the 7H and 4H Mv9kr1/Igri and Asakaze/Manas addition lines in the field in 2012, which increased to 52 and 44 days under short-day illumination in the phytotron. In the winter wheat background, the addition of 2H carrying the photoperiod sensitivity gene Ppd-H1 decreased the flowering time under the short photoperiod regime, but had a very strong delaying effect under field-grown conditions. Considering the yield components under field conditions, 4H was the most fertile of the addition lines, while 7H showed the highest tillering capacity, and Igri 3H had good tillering capacity and the highest number of seeds per plant.     

硬粒小麦-长穗偃麦草附加系、代换系和易位系的创制

通过小麦与长穗偃麦草远缘杂交创制附加系、代换系及易位系是小麦遗传改良中利用长穗偃麦草优良基因的重要途径。本研究将长穗偃麦草特异分子标记、染色体计数、基因组原位杂交(GISH)及非变性原位杂交(ND-FISH)等多种方法相结合,对硬粒小麦Langdon(AABB)与小偃麦8801(AABBEE)的杂交后代群体进行分子细胞学鉴定,创制出硬粒小麦-长穗偃麦草3E、6E染色体双体附加系Du-DA3E和Du-DA6E,硬粒小麦-长穗偃麦草1E(1B)染色体双体代换系Du-DS1E(1B)以及硬粒小麦-长穗偃麦草1AS-1EL染色体易位系Du-T1AS·1EL。创制的4个种质中长穗偃麦草染色体均能稳定遗传,这不仅增加了硬粒小麦-长穗偃麦草附加系和代换系的类型,还为后续利用长穂偃麦草优良基因改良小麦提供了特殊种质资源。     

Substitution of Hordeum marinum ssp. gussoneanum chromosome 7HL into wheat homoeologous group-7

Ditelosomic (Dt) 7HL^mar(7D) and monotelosomic (Mt) 7HL^mar(7A) and 7HL^mar(7B) wheat–barley substitution lines were developed by crossing monosomic 7A, 7B and 7D lines of common wheat cv. Saratovskaya 29 with disomic wheat–barley addition lines (2n = 44) that carry telocentric chromosomes 7HL^mar from Hordeum marinum ssp. gussoneanum 4×. Genomic in situ hybridisation confirmed the presence of barley chromosomes in the wheat genome. The compensating ability of the telosome in each combination was assessed by its transmission rate to progenies of plants with 2n = 41 + t chromosomes. Seed set and transmission rates of the telosome depended on the identity of the competing wheat homoeologue. Of the three chromosomes wheat, the telosome 7HL^mar compensated better for chromosome 7D and poorly for 7B. These and other data are discussed with respect to the phylogenetic relationships between the wheat chromosomes of group 7 and the chromosome of H. marinum, and the practical utility of these lines for wheat improvement is evaluated.     

普通小麦辉县红-荆州黑麦异染色体系的选育及其梭条花叶病抗性鉴定

为发掘和利用荆州黑麦所携抗梭条花叶病基因，综合利用分子细胞遗传学与分子标记技术结合多年抗性鉴定，从高感梭条花叶病小麦地方品种辉县红与荆州黑麦杂交后代（F₇~F₉）中选育出二体异附加系5个(分别添加1R、2R、R3、5R和R7)、5RS端二体异附加系1个和多重异附加代换系2个（染色体组成分别为20’’+2R(2D)’’+4R’’和19’’+1R（1B）’’+2R（2B）’’+4R’’）。鉴定表明，双二倍体荆辉1号高抗梭条花叶病，表明黑麦抗性基因可在小麦背景中稳定表达，2R、R7二体异附加系及2个含2R的多重异附加代换系均表现高抗，推测2R和R7上可能携带抗病基因。这些材料是研究荆州黑麦抗性基因遗传及小麦抗病育种的新种质。     

A New Source of Resistance to Pseudocercosporella herpotrichoides, Cause of Eyespot Disease of Wheat, Located on Chromosome 4V of Dasypyrum villosum

Resistance to Pseudocercosporella herpotrichoides in five wheat cultivars, accession W6 7283 of Dasypyrum villosum, and ‘Chinese Spring’ disomic addition lines of the D. villosum chromosomes IV, 2V, 4V, 5V, 6V and 7V, was evaluated in seedlings by measuring disease progress 6 weeks after inoculation with a β—glucuronidase—transformed strain of the pathogen and by visual estimates of disease severity. D. villosum and the disomic addition line of chromosome 4V were as resistant as wheat cultivars ‘VPM—1’ and ‘Cappelle Desprez’, but less resistant than ‘Rendezvous’. Resistance of the chromosome 4V disomic addition line was equivalent to that of D. villosum.‘Chinese Spring’ and disomic addition lines of IV, 2V, 5V, 6V and 7V were all susceptible. These results confirm Sparaguee's (1936) report of resistance in D. villosum to P. herpotrichoides and establish the chromosomal location for the genes controlling resistance. The presence of chromosome 4V in the addition line and its homocology to chromosome 4 in wheat were confirmed by Southern analysis of genomic DNA using chromosome group 4-specific clones. This genetic locus is not homoeologous with other known genes for resistance to P. herpotrichoides located on chromosome group 7, and thus represents a new source of resistance to this pathogen.     

Chromosome Locations of Additional Barley Enzyme Loci Identified Using Wheat-Barley Addition Lines

Disomic wheat-barley addition lines carrying a full hexaploid set of chromosomes of the wheat cultivar Chinese Spring, plus a pair of either chromosome 1, 2, 3, 4, 6, or 7 from the barley cultivar Betzes, were assayed electrophoretically. Comparisons of zymograms of each addition line with the Chinese Spring wheat and Betzes barky parents of the addition lines gave direct (presumptive in the case of chromosome 5) evidence of the chromosomal locations of nine enzyme loci which have been useful genetic markers in studies of the population genetics, ecogeneties and breeding of barley. Previously reported chromosome locations of several loci were confirmed and evidence was presented for the existence and probable chromosome locations for additional enzyme loci.     

利用SSR鉴定普通小麦-多枝赖草二体异附加系 Line24中外源染色体同源群的归属

用227对小麦微卫星引物进行PCR扩增，76对可在多枝赖草和耐黄矮病的普通小麦-多枝赖草二体异附加系Line24的小麦亲本中国春、丰抗13间检测到多态性。和多枝赖草相同而与Line24其他小麦亲本不同的扩增带。在这76对引物中，发现有4对引物能从Line24中扩增出进一步用Line24和普通小麦杂交得到的7个不同的单体异附加系进行验证，也得到同样的结果，说明这4对微卫星引物扩增出的特异带可以作为Line24中多枝赖草染色体的分子标记。根据这4对引物各自对应的微卫星标记位点在小麦染色体上的位置，说明Line24中附加的一对多枝赖草染色体是第3，5，6和7部分同源群多枝赖草染色体相互易位形成的。     

多小穗小麦品系88F2185抽穗期的染色体效应

采用中国春单体系列对多小穗小麦品系“88F2185”的抽穗期进行了遗传分析。 “88F2185”的早抽穗性状是其3B、 5B和7D染色体的效应。 其中, 7D染色体的效应最强, 5B染色体具早抽穗的新基因。 3B、 5B和7D F2群体中单体株与二体株比较的结果表明, 早抽穗基因是半合、 有效的。     

普通小麦中国春-百萨偃麦草异染色体系的分子标记分析

综合利用HMW-Glu亚基、STS、SSR和RFLP等分子标记对普通小麦中国春、百萨偃麦草、中国春-百萨偃麦草双二倍体和11个中国春-百萨偃麦草异染色体系进行了分析。结果表明，14对SSR、10对STS引物和6个RFLP标记可以特异追踪百萨偃麦草染色质。C7-17及其后代株系C7-17-2等编码百萨偃麦草特异HMW-Glu亚基，添加染色体涉及与小麦第１部分同源群染色体部分同源的1J；1对STS、3对SSR和1个RFLP探针可以特异追踪二体附加系CH05中的百萨偃麦草染色体，并揭示最初根据分带核型确定的J3与小麦第2部分同源群染色体具有较高的部分同源性；2对STS、1个RFLP探针和1对SSR可以追踪CH09的外源染色体，并揭示最初确定的J7与小麦第3部分同源群染色体具有较高的部分同源性；1对STS和1个RFLP探针在CH03、CH04和CH34中具有相同的多态，3个附加系可能添加了相同染色体，最初确定的J₁、J₂和J_?与小麦第7部分同源群染色体具有较高的部分同源性；3对SSR引物可以特异追踪CH12中附加的大片段易位染色体和CH11中的小片段易位染色体，推测易位可能涉及同一条百萨偃麦草染色体。发现13个标记（5个STS、3个RFLP探针和5个SSR）可以追踪未涉及到的4J和5J等染色体。     

Characterization of wheat-alien translocations conferring resistance to diseases and pests: current status

Summary Wild relatives of common wheat, Triticum aestivum, and related species are an important source of disease and pest resistance and several useful traits have been transferred from these species to wheat. C-banding and in situ hybridization analyses are powerful cytological techniques allowing the detection of alien chromatin in wheat. C-banding permits identification of the wheat and alien chromosomes involved in wheat-alien translocations, whereas genomic in situ hybridization analysis allows determination of their size and breakpoint positions. The present review summarizes the available data on wheat-alien transfers conferring resistance to diseases and pests. Ten of the 57 spontaneous and induced wheat-alien translocations were identified as whole arm translocations with the breakpoints within the centromeric regions. The majority of transfers (45) were identified as terminal translocations with distal alien segments translocated to wheat chromosome arms. Only two intercalary wheat-alien transloctions were identified, one induced by radiation treatment with a small segment of rye chromosome 6RL (H25) inserted into the long arm of wheat chromosome 4A, and the other probably induced by homoeologous recombination with a segment derived from the long arm of a group 7 Agropyron elongatum chromosome with Lr19 inserted into the long arm of 7D. The presented information should be useful for further directed chromosome engineering aimed at producing superior germplasm.Contribution No. 96-55-J from the Kansas Experimental Station, Kansas State University, Manhattan, KS 66506-5502, USA.     

Chromosomal locations and genetic relationships of tiller and spike characters in wheat

Number of tillers per plant, plant growth habit in seedling and adult stages, and spike and spikelet characters are agronomically important features of the gross morphology of wheat. To localize to wheat chromosomes the genes for these traits, we scored them in a set of wheat recombinant-inbred mapping lines already well genotyped with molecular markers. Quantitative-trait analysis revealed a region near Gli-A2 (Xpsr10) on the short arm of chromosome 6A strongly affecting tiller number and the correlated trait of seedling growth habit. Genes with opposing effects on adult plant type were localized on the short arms of chromosomes2A and 3A, while genes affecting spike development were assigned to several A- and B-genome chromosomes. None of these genes showed synteny with counterpart QTLs reported to affect the same traits in rice. In the chromosome 2D region containing the photoperiod-insensitivity gene Ppd-D1, the major determinant of heading date in these autumn-sown lines, earliness alleles reduced tiller and spikelet numbers and increased erect seedling growth habit, but showed no influence on adult plant type or spike length. Though several of these morphological traits are generally considered to be associated with winter hardiness and their phenotypic intercorrelations were consistent with the genetic mapping evidence, no association was found between newly identified loci and known vernalization-response or frost-resistance loci. This revised version was published online in August 2006 with corrections to the Cover Date.     

Substituting Ability of Individual Barley Chromosomes for Wheat Chromosomes 1. Substitutions Involving Barley Chromosomes 1, 3 and 6

In order to determine the genetic relatedness of individual barley chromosomes to wheat chromosomes, ‘Betzes’ barley chromosomes 1, 3 and 6 were substituted for individual ‘Chinese Spring’ wheat chromosomes of homoeologous groups 7, 3 and 6, respectively. The substitution status of these lines has been confirmed using isozyme selective markers, chromosome pairing behaviour in F₁ hybrids between the substitution lines and the appropriate double ditelocentric stocks of wheat, and hybridization of cDNA probes to the genomic DNA digests of these substitution lines. Each of the three barley chromosomes provided genetic compensation for the wheat chromosomes they replaced in the substitution plants. From the basis of this compensation with respect to plant vigour and fertility, barley chromosomes 1, 3 and 6 have been designated 7H, 3H and 6H.