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Homologous analysis on centromeric region sequences of A and B chromosomes from rye by FISH

Chinese Science Bulletin 1999年第8期44卷 704-707页

作者： Rongxin Zhang Chengbin Chen Wenqin Song Xiulan Li Ruiyang Chen Nankai Univ Dept Biol Tianjin 300071 Peoples R China

Four centromeric segments from B chromosomes (Bs) of rye have been microdis-sected and amplified by linker adapter PCR (LA-PCR). The PCR products ranged from 100 to 2 000 bp. Fluorescence in situ hybridization (FISH) experiment has been carried out using PCR products as the probe, which was labeled by DIG-11-dUTP. The result confirms that these PCR products from Bs centromeric region are homologous with that of A chromosomes (As) in rye. It also proves that Bs are originated from As.

关键词： B chromosome of rye centromeric region microdissection homology FISH.

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Determination of copy number for 5S rDNA and centromeric sequence RCS2 in rice by Fiber-FISH

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Chinese Science Bulletin 2002年第3期47卷 214-217页

作者： Zongyun Li Siluo Huang Weiwei Jin Shunbin Ning Yunchun Song Lijia Li Wuhan Univ Key Lab Minist Educ Plant Dev Biol Wuhan 430072 Peoples R China

The copy number of 5S rDNA and centromerie sequence RCS2 was determined by extended DNA fiber based fluorescence in situ hybridization (Fiber-FISH) in rice (Oryza sativa ssp. indica cv. Guangluai No. 4) genome. In order to determine the copy number, it is necessary to know the basepair number that a given length DNA fiber contains under a microscope. Therefore, the length of two DNA frag-ments, in which the basepair number had been already known, was measured. The insert sequence of the tested BAC 38D17 was 136 kb and its extended DNA was 56.4 μm long, 2.41 kb/μm on average, while that of the tested BAC 44B4 was 144.5 kb in total and 55.7 μm long, 2.60 kb/μm on average under the microscope. They were very close to the theoretical value of B-DNA in the Watson-Crick DNA model, which is 2.97 kb/μm. According to the average value of basepair number per μm of the two samples mentioned above, that is, 2.51 kb/μm, it could be estimated that the copy number was about 686 for 5S rDNA and 286-1121 for the

关键词： centromeric DNA sequence 5S rDNA copy number fiber based fluorescence in situ hybridization (Fiber-FISH) rice.

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Telomere‑to‑telomere and haplotype‑resolved genome of the kiwifruit Actinidia eriantha

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Molecular Horticulture 2023年第1期3卷 361-371页

作者： Yingzhen Wang Minhui Dong Ying Wu Feng Zhang Wangmei Ren Yunzhi Lin Qinyao Chen Sijia Zhang Junyang Yue Yongsheng Liu School of Horticulture Anhui Agricultural UniversityHefei 230036China School of Forestry Science and Technology Lishui Vocational and Technical CollegeLishui 323000China Ministry of Education Key Laboratory for Bio‑Resource and Eco‑Environment College of Life ScienceState Key Laboratory of Hydraulics and Mountain River EngineeringSichuan UniversityChengdu 610064China

Actinidia eriantha is a characteristic fruit tree featuring with great potential for its abundant vitamin C and strong disease *** has been used in a wide range of breeding programs and functional genomics *** published genome assemblies of *** are quite fragmented and not highly *** multiple sequencing strategies,we get the haplotype-resolved and gap-free genomes of an elite breeding line“Midao 31”(MD),termed MDHAPA and *** new assemblies anchored to 29 pseudochromosome pairs with a length of 619.3 Mb and 611.7 Mb,as well as resolved 27 and 28 gap-close chromosomes in a telomere-to-telomere(T2T)*** on the haplotype-resolved genome,we found that most alleles experienced purifying selection and coordinately *** to the high continuity of assemblies,we defined the centromeric regions of ***,and identified the major repeating monomer,which is designated as *** resource lays a solid foundation for further functional genomics study and horticultural traits improvement in kiwifruit.

关键词： Kiwifruit Actinidia eriantha Genome Telomere-to-telomere Haplotype centromeric

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Synthesis of a Wheat/Maize Hybrid CENH3 Gene, the Genetic Transformation of Wheat, Its Chromosomal Localization and Effects on Chromosome Behaviors in Wheat/Maize Somatic Hybrids

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Agricultural Sciences 2019年第7期10卷 985-1014页

作者： Xiaoyu Yang Jianhui Li Weichang Yu Guangdong Provincial Key Laboratory for Plant Epigenetics College of Life Sciences and Oceanography Shenzhen University Shen-zhen China Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province College of Optoelectronic Engineering Shenzhen University Shenzhen China School of Life Sciences State Key Laboratory of Agrobiotechnology The Chinese University of Hong Kong Hong Kong China

Centromere-specific histone H3 (CENH3) replaces the canonical histone H3 in nucleosomes of functional centromeres, and plays important roles in faithful chromosome segregation during cell division. CENH3 is also important in the recognition of alien centromeres and determines the accommodation or elimination of alien chromosomes in interspecific or intergenic hybridization. In this study, a maize full length CENH3 with a yellow fluorescent protein (YFP) tag at C-terminus (ZmCENH3-YFP) and a synthetic hybrid wmCENH3 with the N-terminus from wheat CENH3 and the histone fold domain (HFD) from maize tagged with a red fluorescent protein (RFP) at the C-terminus (wmCENH3-RFP) were transformed to wheat by biolistics transformation. Transgenic wheat plants with both ZmCNEH3-YFP and wmCENH3-RFP genes were identified by PCR. The expression of ZmCENH3-YFP was not observed, while the expression of wmCENH3-RFP could be detected by RT-PCR, direct fluorescence microscopy, and immunostaining with anti-RFP antibody. The expressed wmCENH3-RFP was localized to nuclei as dotted patterns, indicating its targeting to wheat centromeres. Somatic hybridization was performed between wmCENH3-RFP transgenic wheat and transgenic maize that expressed a ZmCENH3-YFP gene to investigate chromosome behaviors in somatic hybrids. Cytological and FISH analyses of somatic hybrid cells showed the formation of micronuclei and lagging chromatin in both somatic hybridizations with or without the wmCENH3-RFP transgene, indicating that ectopically expressed wmCENH3 could not overcome chromosome elimination in wheat/maize somatic hybrids. Immunostaining of wmCENH3-RFP and ZmCENH3-YFP in early stage somatic hybrid cells indicated that both wmCENH3-RFP and ZmCENH3-YFP proteins were expressed, but their binding patterns changed from the commonly observed dotted patterns to diffused ones, suggesting that the inactivation of CENH3 might be a factor for chromosome elimination in wheat/maize somatic hybridization.

关键词： centromeric Histone H3 Genetic Transformation Chromosome Elimination Somatic Hybridization Wheat Maize

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