小麦及其近缘种中基因组特异性DNA重复序列的研究进展

本文对小麦族植物中基因组特异性DNA重复序列的分类、基本特征、分离和鉴定方法、在小麦遗传改良中的应用以及未来研究的发展趋势进行了简述。综合已有的研究结果可以看出基因组特异性DNA重复序列是小麦族植物基因组特异性形成的重要构成部分。对基因组特异性DNA重复序列的研究是认识小麦族植物基因组的有效途径之一,基因组特异性DNA重复序列的应用将进一步促进小麦族植物分子细胞遗传学和普通小麦遗传改良研究的进展。 Advances in Studies of Genome-Specific Repetitive DNA Sequences in Wheat and Related Species BAI Jian-rong1,2,JIA Xu1,WANG Dao-wen1 1.The State Key Laboratory of Plant Cell and Chromosome Engineering,Institute of Genetics and Developmental Biology,The Chinese Academy of Sciences,Beijing 100101,China; 2.Crop Genetics Institute,Shanxi Academy of Agricultural Sciences,Taiyuan 030031,China Abstract:In this paper we review recent advances in studies of several aspects of genome specific repetitive DNA sequences in wheat and related species.The available results demonstrate that genome specific repetitive DNA sequences are important components of genome specificity in wheat and related species.Research on genome specific repetitive DNA sequences is essential to the elucidation of genome function.The application of genome specific repetitive DNA sequences will aid molecular cytogenetic studies in wheat and related species and contributes to genetic improvement of common wheat. Key words：wheat;genome specific repetitive DNA sequence;chromosome     

Synthetic hexaploid wheat and its utilization for wheat genetic improvement in China

Synthetic hexaploid wheat （Triticum turgidum x Aegilops tauschii） was created to explore for novel genes from T. turgidum and Ae. tauschii that can be used for common wheat improvement. In the present paper, research advances on the utilization of synthetic hexaploid wheat for wheat genetic improvement in China are reviewed. Over 200 synthetic hexaploid wheat （SHW） accessions from the International Maize and Wheat Improvement Centre （CIMMYT） were introduced into China since 1995. Four cultivars derived from these, Chuanmai 38, Chuanmai 42, Chuanmai 43 and Chuanmai 47, have been released in China. Of these, Chuanmai 42, with large kernels and resistance to stripe rust, had the highest average yield （〉 6 t/ha） among all cultivars over two years in Sichuan provincial yield trials, outyielding the commercial check cultivar Chuanmai 107 by 22,7%. Meanwhile, by either artificial chromosome doubling via colchicine treatment or spontaneous chromosome doubling via a union of unreduced gametes （2n） from T. turgidum-Ae, tauschii hybrids, new SHW lines were produced in China. Mitotic-like meiosis might be the cytological mechanism of spontaneous chromosome doubling. SHW lines with genes for spontaneous chromosome doubling may be useful for producing new SHW-alien amphidiploids and double haploid in wheat genetic improvement.     

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.     

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.     

Acta Genetica Sinica and HEREDITAS（Beijing）

Acta Genetica Sinica （Yichuan Xuebao） （IS SN 0379-4172） （CN 11-1914/R） and HEREDITAS （BeiJing）/ （Yichuan） （ISSN 0253-9772, CN11-1913/R） are sponsored by the Genetics Society of China, and the Institute of Genetics and Developmental Biology of Chinese Academy of Sciences,     

Development of Triticum aestivum-Leymus racemosus translocation lines using gametocidal chromosomes

Specific chromosomes of certain Aegilops species introduced into wheat genome background may often facilitate chromosome breakage and refusion, and finally result in a variety of chromosome restructuring. Such a phenomenon is commonly called gametocidal effect of the chromosomes. The chromosome 2C of Ae. cylindrica is one of such chromosomes. In the present study, scab resistant wheat-L. racemosus addition lines involving chromosomes Lr.2 and Lr.7 were crossed to wheat-Ae. cylindrica disomic addition line Add2C. Then F₁ hybrids were subsequently backcrossed with wheat cv “Chinese Spring”. BC₁ plants with chromosome structural aberration were identified by C-banding. In the self-pollinated progenies of these plants, three translocation lines were developed and characterized by mitotic and meiotic analysis combined with C-banding and fluorescent in situ hybridization (FISH) using biotin-labeled genomic DNA of L. racemosus as probe. Some other putative translocation lines to be further characterized were also found. The practicability and efficiency of the translocation between wheat and alien chromosomes induced by gametocidal chromosomes, as well as the potential use of the developed alien translocation lines were also discussed.     

Induction of small-segment-translocation between wheat and rye chromosomes

A new approach to produce wheat-rye translocation, based on the genetic instability caused by monosomic addition of rye chromosome in wheat, is described. 1 283 plants from the selfed progenies of monosomic addition lines with single chromosome of inbred rye line R12 and complete chromosome complement of wheat cultivar Mianyang 11 were cytologically analyzed on a plant-by-plant basis by the improved C-banding technique. 63 of the plants, with 2n = 42, were found containing wheat-rye translocation or substitution, with a frequency of 4. 91% . Compared with the wheat parent, other 32 plants with 2n = 42 exhibited obvious phenotypic variation, but their com-ponent of rye chromosome could not be detected using the C-banding technique. In situ hybridization with a biotin-la-beled DNA probe was used to detect rye chromatin and to determine the insertion sites of rye segments in the wheat chromosomes. In 20 out of the 32 variant wheat plants, small segments of rye chromosomes were found being inserted into dif     

Molecular Cytogenetic Characterization of a Wheat - Leymus mollis 3D（3Ns） Substitution Line with Resistance to Leaf Rust

Leymus mollis （Trin.） Pilger （NsNsXmXm, 2n = 28）, a wild relative of common wheat, possesses many potentially valuable traits that could be transferred to common wheat during breeding programs. In this study, the karyotypic constitution of a wheat - L. mollis 3D（3Ns#1） disomic substitution line isolated from the F5 progeny of octoploid Tritileymus M842-16 x Triticum durum cv. D4286, which was designated as 10DM57, was determined using genomic in situ hybridization （GISH）, fluorescent in situ hybridization （FISH）, SSR markers, and EST- STS markers. Screening of mitosis and meiosis showed that 10DM57 had a chromosome karyotype of 2n = 42 =21Ⅱ. GISH indicated that 10DM57 was a line with 40 chromosomes from wheat and two of the Ns chromosomes from L. mollis, which formed a ring bivalent in pollen mother cells at metaphase I. FISH analysis showed that the chromosome 3D may be replaced by 3Ns#1 in 10DM57. DNA markers, including SSR and EST-STS primers, showed that the pair of wheat chromosome 3D in 10DM57 was substituted by the pair of chromosome 3Ns#t from L. mollis. Evaluation of the agronomic traits showed that, compared with its common wheat relative 7182, 10DM57 was resistant to leaf rust while the spike length and number of spikes per plant were improved significantly, which correlated with a higher wheat yield. The new germplasm, 10DM57, could be exploited as an intermediate material in wheat genetic and breeding programs.     

Physical mapping of three fruit ripening genes：Endopolygalacturonase,ACC oxidase and ACC synthase from apple（Malus x domestica） in an apple rootstock A106（Malus sieboldii

The apple rootstock,A106(Malus sieboldii),had 17 bivalents in pollen mother cells at meiotic metaphase 1,and 17 chromosomes in a haploid pollen cell.Karyotypes were prepared from root-tip cells with 2n=34 chromosomes,Seven out of 82 karyotypes(8.5%) showed one pari of satellites at the end of the short arm of chromosome 3.C-bands were shown on 6 pairs of chromosomes 2,4,6,8,14,and 16 near the telomeric regions of short arms.Probes for three ripening-related genes from Malus x domestica:endopolygalacturonase(EPG,0.6kb),ACC oxidase(1.2kb),and ACC synthase(2kb)were hybridized in situ to metaphase chromosomes of A106.Hybridization sites for the EPG gene were observed on the long arm of chromosome 14 in 15 out of 16 replicate spreads and proximal to the centromere of chromosomes 6 and 11.For the ACC oxidase gene,hylridization sites were observed in the telomeric region of the short arm of chromosomes 5 and 11 in 87% and 81% of 16 spreads respectively,proxiaml to the centromere of chromosome 1 in 81% of the spreads,and on the long arm of chromosome 13 in 50% of the spreads. Physical mapping of three fruit ripening genes in an apple rootstock A106.Twenty five spreads were studied for the ACC synthase gene and hybridization sites were observed in the telomeric region of the short arm of chromosome 12 in 96% of the spreads.chromosomes 9 and 10 in 76% of the spreads,and chromosome 17 in 56% of the spreads.     

Wide hybridization: engineering the next leap in wheat yield

Wide hybridization and chromosome engineering constitute an active and important area of plant biology research. The insights obtained in this field have contributed significantly to the understanding of the structure and function of plant chromosomes and genomes, and to the genetic improvement of many agricultural crops. Owing to well defined genomic relationships, clearly visible metaphase chromosomes, and relatively high interspecific and intergeneric crossability, wheat and related Triticeae species form ideal experimental systems for conducting wide hybridization and chromosome engineering studies （Sears, 1972, 1981; Singh and Jauhar, 2006）. Pioneering investigations have demonstrated that the entire genome or chromatin from Triticeae relatives can be transferred into wheat genome through chromosome engineering （re- viewed by Qi et al., 2007）.     

Insecticide resistance status and detoxification enzymes of wheat aphids Sitobion avenae and Rhopalosiphum padi

正Dear Editor,Wheat aphids are serious pests in wheat growing areas of China,and can lead to from 10%to over 30%reduction in wheat production(Wang et al.,2006).The main wheat aphids are Sitobion avenae(Fabricius),Rhopalosiphum padi(Linnaeus),Metopolophium dirhodum(Walker),and Schizaphis graminum(Rondani).In the Huang-Huai area of China,the dominant wheat aphid species are S.avenae and R.padi.For many years,farmers have applied chemical pesticides to control wheat aphids.The number of wheat aphids in north     

Development of Triticum aestivum-Leymus racemosus translocation lines using gametocidal chromosomes

Maan[1] and Endo[2] et al. first reported that some chromosomes from Ae. longgissima, Ae. sharonensis and Ae. triuncialis showed preferential transmission when introduced into wheat background. The mechanism for this phenomenon rests with the fact that contrary to the normal fertility of gametes with these chromosomes, chromosome structural aberrations occur seriously in the gametes without these chromosomes, causing less compatibility in selective fertilization and resulting in semi-sterilit…     

对水稻第9和第12染色体编号分歧的细胞学考证

在水稻细胞遗传研究中, 对于染色体编号有着较多的争议,这在几条长度较短的染色体上显得尤为突出。为有比较地研究这几条染色体在水稻染色体组中的正确编号,本研究以涉及两条较短染色体相互易位的易位杂合体RT9-12为材料,分析了易位系与普通品种日本晴减数分裂粗线期染色体的形态特征。结果表明,该易位系的易位染色体并非第9和第12染色体,而是第10和第11染色体,从而认为目前国际上统一编号的第9、12染色体,根据染色体的实际长度可能分别为第10、11染色体。 Abstract:Rice chromosomes in mitosis are usually too small to be identified clearly one from others.In recent years,pachytene chromosomes in meiosis have been in vestigated intensively for establishing unified numbering system.However,divergence in numbering system is still existing especially for some short chromosomes such as chromosome 9 and 12.In order to verify these chromosomes,a translocation line RT9-12 and a japonica variety Nipponbare were carefully investigated for all the chromosomes morphologically in late pachytene stage.It was found that the chromosomes involved in translocation were chromosome 10 and 11 in stead of chromosome 9 and 12 as being compared with the karyotype of Nipponbare.So we consider that the chromosome 9 and 12 in the present rice chromosome numbering system could be chromosome 10 and 11 according to their length,arm ratio and the relationship with nucleolus.     

Variation of B Chromosome Associated with Tissue Culture in Wheat-rye Cross

In vitro variation of B chromosomes was studied by examining the callus cells derived from the immature embryos from a cross of Chinese Spring wheat （Triticum aestivum L.） and Fin 7416 rye （Secale cereale L.） carrying two B chromosomes. In 40-d-old callus cells, the numbers of B chromosomes ranged from one to four in 65.6% of the cells observed. The distribution of B chromosome numbers was associated with the ploidy levels of the normal chromosomes （A chromosomes）. The frequency of the cells with high numbers of B chromosomes （i.e., three or four B chromosomes） in the amphiploid cells with 56 A chromosomes was greater than those in the haploid cells with 28 A chromosomes. Although structural changes in the rye A chromosomes were observed, cytological observation and genomic in situ hybridization demonstrated that the rye B chromosomes were conserved in morphological appearance following tissue culture.     

中国特有植物金铁锁的细胞学研究

In this paper, Psammosilene tunicoides, an endemic species to China, was cytologically studied for the first time. The morphology of the nuclei at resting stage was categorized to be simple chromocentre type. The morphology of mitofic-pmphase chromosomes was categorized to be the interstitial type.28 chromosomes were observed at the mitotic metaphase, and 14 bivalent chromosomes were observed at diakinesis. So, the basic chromosome number was confirmed to be x = 14. Psomzr~silene tunicoides is different from Silene rubicunda in the basic chromosome number and the morphology of the nuclei at resting stage and mitotic-prophase chromosomes, because Sgene rub/cund has the basic chromosome number of x= 10 and 12, and its nuclei at resting stage and mitotic-prophase chromosomes is sparsely diffuse type and continuous type respectivrly.     

小麦基因组研究进展

本文从小麦遗传图谱、物理图谱、比较基因组、基因组测序和EST 5个方面,介绍国内外小麦基因组的研究进展。我们利用W7984×Opata重组近交系的RFLP作图群体,对33个与小麦水分利用效率有关的性状进行了QTL遗传图谱比较分析,结果显明：在第一部分同源群染色体（1A,1B）上的着丝粒周围,分布有控制光合作用和根系特性的基因簇。在第二部分同源染色体上,有控制单株水分利用效率、根冠形态和生长发育的基因簇存在。在第六部分同源染色体上,6A和6B上都分别有由控制根系多个QTL组成的基因簇,6D染色体着丝粒周围有一个大的基因簇,由7个控制叶片和单株水分利用效率的QTL组成,说明第六部分同源染色体在小麦水分利用效率遗传方面起重要作用。 Abstract:Research development of genetic mapping,physics mapping,genome sequencing and expressed sequence tags in wheat have been reviewed in this paper.RFLP genetic linkage map of wheat recombinant inbred lines derived from W7984×Opata,was used to study QTL of 33 traits associated with water use efficiency.Compared with QTL map of 7 group homeologues chromosomes,the results were showed as follows:nearby the centromeric region of 1A and 1B chromosome,the gene cluster of controlling photosynthetic and root traits were located.The gene clusters of controlling water use efficiency per plant,root and plant height and growth rate were located on the 2 group chromosomes.The gene clusters of controlling root traits were located on the 6A an 6B chromosome,there was a big gene cluster mad up by 7 QTLs controlling water use efficiency of wheat leaf and per plant nearby the centromeric region of 6D chromosome.It showed that 6th homeologous chromosomes play an important role in controlling water use efficiency in wheat.     

Thinopyrum ponticum and Th. intermedium:the promising source of resistance to fungal and viral diseases of wheat

Thinopyrum ponticum and Th. intermedium provide superior resistance against various diseases in wheat （Ttricum aestivum）. Because of their readily crossing with wheat, many genes for disease resistance have been introduced from the wheatgrasses into wheat. Genes for resistance to leaf rust, stem rust, powdery mildew, Barley yellow dwarf virus, Wheat streak mosaic virus, and its vector, the wheat curl mite, have been transferred into wheat by producing chromosome translocations. These genes offer an opportunity to improve resistance of wheat to the diseases; some of them have been extensively used in protecting wheat from damage of the diseases. Moreover, new resistance to diseases is continuously detected in the progenies of wheat-Thinopyrum derivatives. The present article summaries characterization and application of the genes for fungal and viral disease-resistance derived from Th. ponticum and Th. intermedium.     

Identification and Validation of a Major Quantitative Trait Locus for Slow-rusting Resistance to Stripe Rust in Wheat

Stripe (yellow) rust,caused by Puccinia striiformis Westend.f.sp.tritici Eriks (Pst),is one of the most important wheat (Triticum aestivum L.) diseases and causes significant yield losses.A recombinant inbred (RI) population derived from a cross between Yanzhan 1 and Xichang 76-9 cultivars was evaluated for resistance to wheat stripe rust strain CYR32 at both the seedling and adult plant stages.Four resistance quantitative trait loci (QTLs) were detected in this population,in which the major one,designated as Yrq1,was mapped on chromosome 2DS.The strategy of using the Brachypodium distachyon genome,wheat expressed sequence tags and a draft DNA sequences (scaffolds) of the D-genome (Aegilops tauschii Coss.) for the development of simple sequence repeat (SSR) markers was successfully used to identify 147 SSRs in hexaploid wheat.Of the 19 polymorphic SSRs in the RI population,17 SSRs were mapped in the homeologous group 2 chromosomes near Yrq1 region and eight SSRs were genetically mapped in the 2.7 cM region of Yrq1,providing abundant DNA markers for fine-mapping of Yrq1 and marker-assisted selection in wheat breeding program.The effectiveness of Yrq1 was validated in an independent population,indicating that this resistance QTL can be successfully transferred into a susceptible cultivar for improvement of stripe rust resistance.     

Mapping of a BYDV resistance gene from Thinopyrum intermedium in wheat background by molecular markers

The wheat line H960642 is a homozygous wheat-Thinopyrum intermedium translocation line with resistance to BYDV by genomie in situ hybridization (GISH) and RFLP analysis. The genomie DNA of Th. intermedium was used as a probe, and eonunon wheat genomie DNA as a blocking in GISH experiment. The results showed that the chromosome segments of Th. intermedium were transferred to the distal end of a pair of wheat chromosomes. RFLP analysis indicated that the transloeation line H960642 is a T7DS·7DL-7XL translocation by using 8 probes mapped on the homoeologous group 7 in wheat. The tranalocation breakpoint is located between Xpsr680 and Xpsr965 about 90—99 cM from the centromere. The RFLP markers psr680 and psr687 were closoly linked with the BYDV resistance gene. The gene is located on the distal end of 7XL around Xpsr680 and Xpsr687.     

Microdissection of additional chromosome in common wheat-Th. intermedium TAI-27 and screening of its special probe

It was identified that there were 4 St chromosomes derived from Th, intermedium in common wheat-Th. intermedium alien additional line TAI-27 by in situ hybridization. Two St chromosomes added to wheat chromosome in TAI-27 as well as two of them replaced two of 42 in wheat chromosomes. This indicates that TAI-27 is not only an alien additional line, but also a replacing line. The additional chromosomes in TAI-27 were microdissected and a microcloning library was constructed. A special probe of Th. intermedium was obtained from a microcloning library. The sequence analysis indicated that there were no homology with Genebank data. This makes it possible to screen genes with the disease-resistance, adversity-tolerance and fine character from Th.intermedium.