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Analysis of the meiosis in the F_1 hybrids of Longiflorum × Asiatic(LA) of lilies(Lilium) using genomic in situ hybridization

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Journal of Genetics and genomics 2008年第11期35卷 687-695页

作者： Shujun Zhou Munikote S. Ramanna Richard G.E Visser Jaap M. van Tuyl Institute of Landscape College of Agriculture ＆ Biotechnology Zhejiang University Hangzhou 310029 China Plant Breeding Lab Wageningen University P.O. Box 386 6700 AJ Wageningen The Netherlands

Longiflorum and Asiatic lilies of the genus Lilium of the family Liliaceae are two important groups of modem lily cultivars. One of the main trends of lily breeding is to realize introgression between these groups. With cut style pollination and embryo rescue, distant hybrids between the two groups have been obtained. However, the FI hybrids are highly sterile or some of them could produce a small number of 2n gametes, and their BC1 progenies are usually triploids. Dutch lily breeders have selected many cultivars from these BC1 progenies based on their variation. It is presumably suggested that such variation could be caused by intergenomic recombination and abnormal meiosis during gamete formation in F1 hybrids of Longiflorum × Asiatic （LA） hybrids in Lilium. Therefore, the meiotic process of ten F1 LA hybrids was cytologically investigated using genomic in situ hybridization and traditional cytological methods in the present research. The results showed that： at metaphase I, the homoeologous chromosome pairing among different F1 hybrids ranged from 2.0 to 11.4 bivalents formed by homoeologous chromosomes per pollen mother cell （PMC）, and very few multivalents, and even very few bivalents were formed by two chromosomes within one genome rather than homoeologous chromosomes in some PMCs; at anaphase I, all biva- lents were disjoined and most univalents were divided. Both the disjoined bivalents （half-bivalents） and the divided univalents （sister chromatids） moved to the opposite poles, and then formed two groups of chromosomes; because the two resulting half-bivalents retained their axes in the cell undisturbed, many crossover types, including single crossovers, three strand double crossovers, four strand double crossovers, four strand triple crossovers, and four strand multiple crossovers between the non-sister chromatids in the tetrads of bivalents, were clearly inferred by analyzing the breakpoints on the disjoined bivalents. The present investigation not only ex

关键词： Lilium genomic in situ hybridization abnormal meiosis crossover 2n gamete

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Cytogenetic comparisons between A and G genomes in Oryza using genomic in situ hybridization

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Cell Research 2006年第3期16卷 260-266页

作者： Zhi Yong Xiong~(1,2) Guang Xuan Tan~1 Guang Yuan He~2 Guang Cun He~1 Yun Chun Song~11 Key Laboratoty of the Ministry of Education for Plant Developmental Biology,College of Life Sciences,Wuhan University,Wuhan 430072,China.2 China-UK HUST-R Res Genetic Engineering and genomics Joint Laboratory,Huazhong University of Science and Technology,Wuhan 430074,China Key Laboratory of the Ministry of Education for Plant Developmental Biology College of Life Sciences Wuhan University Wuhan China China-UK HUST-R Res Genetic Engineering and genomics Joint Laboratory Huazhong University of Science and Technology Wuhan China

The genomic structures ofOryza sativa （A genome） and *** （G genome） were comparatively studied usingbieolor genomic in situ hybridization （GISH）.GISH was clearly able to discriminate between the chromosomes of *** and *** in the interspecific F1 hybrids without blocking DNA,and co-hybridization was hardly *** average mitotic chromosome length of *** was found to be 1.69 times that of ***.A comparison of4,6-diamidino-2-phenylindole staining showed that the chromosomes of *** were more extensively labelled,suggesting that the G genome is amplified with more repetitive sequences than the A *** interphase nuclei,9-12chromocenters were normally detected and nearly all the chromocenters constituted the G genome-specific *** larger chromocenters formed by chromatin compaction corresponding to the G genome were detected in the hybridcompared with its *** pachytene of the F1 hybrid,most chromosomes of A and G did not synapse eachother except tot 1-2 chromosomes paired at the end of their *** meiotic metaphase I,three types of chromosomalassociations,i.e.O,sativa-O,sotiva（A-A）,***-O,meyeriana （A-G） and ***-O,meyeriana （G-G）,wereobserved in the F1 *** A-G chromosome pairing configurations included bivalcnts and *** resultsprovided a foundation toward studying genome organization and evolution of ***.

关键词： Oryza sativa Oryza meyeriana 45S rDNA genomic in situ hybridization

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genomic in situ hybridization analysis on intergeneric hybrids between Brassica species and Orychophragmus violaceus

Genomic in situ hybridization analysis on intergeneric hybri...

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第一届亚洲植物染色体学术讨论会

作者： M. Ceccarelli S. Minelli A.Contento P.G.Cionini Department of Cellldlar and Molecular Biology University of Perugia Via A . Pascoli I-06123 Perugia Italy Department of Cellldlar and Molecular Biology University of Perugia Via A . Pascoli I-06123 Perugia Italy Department of Cellldlar and Molecular Biology University of Perugia Via A . Pascoli I-06123 Perugia Italy Department of Cellldlar and Molecular Biology University of Perugia Via A . Pascoli I-06123 Perugia Italy

＜正＞ In sexual intergeneric hybrids between the six cultivated Brassica species and Orychophragmus violaceus , the possible occurrence of both complete and partial separation of the parental genomes during mitosis and meiosis has been proposed and the interrelationships among the cytogenetics of these hybrids have been shown. To confirm and further elucidate the findings

关键词： Brassica species Orychophragmus violaceus Intergeneric hybrids Genome separation genomic in situ hybridization

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Genetic Relationships Among Five Basic Genomes St, E, A, B and D in Triticeae Revealed by genomic Southern and in situ hybridization

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Journal of Integrative Plant Biology 2007年第7期49卷 1080-1086页

作者： Zhao Liu Dayong Li Xueyong Zhang Key Laboratory of Crop Germplasm and Biotechnology Institute of Crop Sciences Chinese Academy of Agricultural Sciences Beijing 100081 China Institute of Genetics and Developmental Biology the Chinese Academy of Sciences Beijing 100101 China.

The St and E are two important basic genomes in the perennial tribe Triticeae （Poaceae）. They exist in many perennial species and are very closely related to the A, B and D genomes of bread wheat （Triticum aestivum L.）. genomic Southern hybridization and genomic in situ hybridization （GISH） were used to analyze the genomic relationships between the two genomes （St and E） and the three basic genomes （A, B and D） of T. aestivum. The semi-quantitative analysis of the Southern hybridization suggested that both St and E genomes are most closely related to the D genome, then the A genome, and relatively distant to the B genome. GISH analysis using St and E genomic DNA as probes further confirmed the conclusion. St and E are the two basic genomes of Thinopyrum ponticum （StStE^eE^bE^x） and Th. intermedium （StE^eE^b）, two perennial species successfully used in wheat improvement. Therefore, this paper provides a possible answer as to why most of the spontaneous wheat-Thinopyrum translocations and substitutions usually happen in the D genome, some in the A genome and rarely in the B genome. This would develop further use of alien species for wheat improvement, especially those containing St or E in their genome components.

关键词： E genome fluorescent in situ hybridization genomic in situ hybridization perennial Triticeae St genome wheat,

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

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Journal of Integrative Plant Biology 2006年第7期48卷 838-847页

作者： Diao-Guo An Li-Hui Li Jun-Ming Li Hong-Jie Li Yong-Guan Zhu Centre of Agricultural Resources Institute of Genetics and Developmental Biology the Chinese Academy of Sciences Shijiazhuang 050021 China Institute of Crop Sciences the Chinese Academy of Agricultural Sciences Beijing 100081 China Research Center for Eco-environmental Sciences the Chinese Academy of Sciences Beijing 100085 China Graduate School of the Chinese Academy of Sciences Beijing 100039 China

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.

关键词： genomic in situ hybridization nullisomic back-cross procedure PCR powdery mildew resistance wheat-rye chromosome substitution line

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

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Journal of Integrative Plant Biology 2009年第9期51卷 834-839页

作者： Bohong Tian Hongjie Li Cangzhou Academy of Agricultural and Forestry Sciences Cangzhou 061001 China The National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI) Institute of Crop Sciences the Chinese Academy of Agricultural Sciences Beijing 100081 China

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.

关键词： amphiploid B chromosome genomic in situ hybridization Secale cereale tissue culture Triticum aestivum.

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Identification and cell wall analysis of interspecific hybrids between Oryza sativa and Oryza ridleyi

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Journal of Integrative Agriculture 2017年第8期16卷 1676-1681页

作者： YI Chuan-deng LI Wei WANG De-rong JIANG Wei HU Da-bang ZHOU Yong LIANG Guo-hua GUMing-hong Jiangsu Key Laboratory of Crop Genetics and Physiology/Co-Innovation Center for Modern Production Technology of Grain Crops Key Laboratory of Plant Functional Genomics of Ministry of EducationYangzhou UniversityYangzhou 225009P.R.China

Oryza ridleyi is an allotetraploid wild species with the HHJJ genome, and Oryza sativa is a diploid cultivated rice that has the AA genome. Although the wide hybrid between the two species is difficult to obtain, we overcome this difficulty by young embryo rescue. An obvious heterosis was primarily found for the plant height, tillering ability, vegetative vigor, etc. However, the hybrid panicle and culm traits were found to resemble that of the wild rice parent, O. ridleyi, for the long awns, exoteric purple stigma, grain shattering, dispersed panicles, and culm mechanical strength. genomic in situ hybridization （GISH） analysis was subsequently performed on the mitotic metaphase chromosome of the root tips, and we determined that the hybrid is an allotriploid with 36 chromosomes and its genomic constitution is AHJ. Chemical analyses conducted on the culm of O. sativa, O. ridleyi, and their interspecific hybrids showed that major changes occurred in the xylose, glucose, and arabinose concentrations, which are correlated with the specific hemicellulose polymer and cellulose components that are important in the primary cell walls of green plants. Meanwhile, the culm anatomical analyses indicated that additional large vascular bundles and an extra sclerenchyma cell layer were found in O. ridleyi. Additionally, further thickening of the secondary cell walls of the cortical fiber sclerenchyma cells and the phloem companion cells was discovered in O. ridleyi and in the interspecific hybrids. These results imply that there may be a potential link between culm mechanical strength and culm anatomical structure.

关键词： Oryza sativa Oryza ridleyi interspecific hybrid genomic in situ hybridization cell wall

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Induction and transmission of wheat-Haynaldia villosa chromosomal translocations

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Journal of Genetics and genomics 2009年第5期36卷 313-320页

作者： Yaping Cao Tongde Bie Xiue Wang Peidu Chen National Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics Institute. Nanjing Agricultural University Nanjing 210095 China Wheat Research Institute Shanxi Academy of Agricultural Sciences Linfen 041000 China Yangzhou Academy of Agricultural Sciences Yangzhou 225007 China

In order to develop more wheat-Haynaldia villosa translocations involving different chromosomes and chromosome segments of H. villosa, T. durum-H, villosa amphiploid was irradiated with ^60Co γ-rays at doses of 800, 1,200, and 1,600 rad. Pollen collected from the spikes 1, 2, and 3 days after irradiation were transferred to emasculated spikes of the common wheat cv. ‘Chinese Spring＇. genomic in situ hybridization was used to identify wheat-H, villosa chromosome translocations in the M1 generation. Transmission of the identified translocation chromosomes was analyzed in the BC1, BC2, and BC3 generations. The results indicated that all three irradiation doses were highly efficient for inducing wheat-alien translocations without affecting the viability of the M1 seeds. Within the range of 800-1,600 rad, both the efficiency of translocation induction and the frequency of interstitial chromosome breakage-fusion increased as the irradiation dosage increased. A higher translocation induction frequency was observed using pollen collected from the spikes 1 day after irradiation over that of 2 or 3 days after irradiation. More than 70% of the translocations detected in the M1 generation were transmitted to the BC1 through the female gametes. All translocations recovered in the BC1 generation were recovered in the following BC2, and BC3 generations. The transmission ability of different translocation types in different genetic backgrounds showed an order of ‘whole-arm translocation 〉 small alien segment translocation 〉 large alien segment translocation＇, through either male or female gametes, In general, the transmission ability through the female gametes was higher than that through the male gametes. By this approach, 14 translocation lines that involved different H. villosa chromosomes have been identified in the BC3 using EST-STS markers, and eight of them were homozygous.

关键词： Triticum durum-Haynaldia villosa amphiploid pollen irradiation chromosomal translocation translocation transmission genomic in situ hybridization

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Microdissection of Haynaldia villosa Telosome 6VS and Cloning of Species-specific DNA Sequences

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Acta Botanica Sinica 2002年第3期44卷 307-313页

作者：孔凡晶陈孝马有志辛志勇李连成张增艳林志姗中国农业科学院作物育种栽培研究所

The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6VS was microdissected with a needle and transferred into a 0.5 mL Ep tube. In the 'single tube', all the subsequence steps were conducted. After two round of LA (Linker adaptor)_PCR amplification, the size of PCR bands ranged from 100 to 3 000 bp, with predominate bands 600-1 500 bp. The products were confirmed by Southern blotting analysis using Haynaldia villosa (L.) Schur. genomic DNA labeled with 32 P as probe. The PCR products were purified and ligated into clone vector-pGEM_T easy vector. Then, the plasmids were transformed into competence E. coli JM109 with cool CaCl 2. It was estimated that there were more than 17 000 white clones in the library. The size of insert fragments distributed from 100-1 500 bp, with average of 600 bp. Using H. villosa genomic DNA as probe, dot blotting results showed that 37% clones displayed strong and medium positive signals, and 63% clones had faint or no signals. It is demonstrated that there were about 37% repeat sequence clones and 67% single/unique sequence clones in the library. Eight H. villosa_specific clones were screened from the library, and two clones pHVMK22 and pHVMK134 were used for RFLP analysis and sequencing. Both of them were H. villosa specific clones. The pHVMK22 was a unique sequence clone, and the pHVMK134 was a repeat sequence clone. When the pHVMK22 was used as a probe for Southern hybridization, all the powdery mildew resistance materials showed a special band of 2 kb, while all the susceptible ones not. The pHVMK22 may be applied to detect the existence of Pm21.

关键词： microdissection and microcloning of chromosome Haynaldia villosa genomic in situ hybridization alien substitution of telosome species_specific DNA sequences RFLP

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Structural Changes of 2V Chromosome of Haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

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Agricultural Sciences in China 2008年第7期7卷 804-811页

作者： CHEN Quan-zhan CAO Ai-zhong QI Zeng-jun ZHANG Wei CHEN Pei-du National Key Laboratory of Crop Genetics and Germplasm Enhancement/Nanjing Agricultural University Nanjing 210095 P.R. China Genetics Laboratory Life Science Department Nanjing Xiaozhuang University Nanjing 211171 P.R. China

Haynaldia villosa （2n=2X= 14, VV）, a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetic Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge （a remarkable morphology） mapped on 2V of Haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V（2D） and the hybrid F1 was then self-crossed. Chromosome C-banding, genomic in situ hybridization （GISH）, and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids Fz and F3. To date, four translocations including one small segmental translocation T6BS·6BL-2VS, two whole arm translocations （preliminarily designed as T3DS·2VL and T2VS.7DL） and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one isochromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25.120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 （2BL） tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS-7DL, Mt2VS, 2VS-2VS as well as the parent DS2V（2D）, whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C ofAegilops triuncialis could successfully induce chromosome 2V structural changes.

关键词： Triticum aestivum Haynaldia villosa gametocidal chromosome 3C chromosome aberration C-banding genomic in situ hybridization molecular marker

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