【目的】深化柑橘减数分裂粗线期染色体结构组成和配对分析。【方法】首次利用柑橘染色体全长特异寡核苷酸荧光原位杂交(oligo-fluorescence in situ hybridization,oligo-FISH)涂染探针,精准示踪鉴定沙田柚粗线期1号染色体的结构特征,...【目的】深化柑橘减数分裂粗线期染色体结构组成和配对分析。【方法】首次利用柑橘染色体全长特异寡核苷酸荧光原位杂交(oligo-fluorescence in situ hybridization,oligo-FISH)涂染探针,精准示踪鉴定沙田柚粗线期1号染色体的结构特征,比较其与有丝分裂间期细胞核、有丝分裂前中期和中期染色体的结构差异,并揭示粗线期全长染色体的同源配对。【结果】观测到同源配对的全长粗线期与有丝分裂中期较大染色质结构差异。经测定,柚粗线期1号染色体平均全长、长臂长、短臂长和臂比分别为中期的13.93倍、16.54倍、10.44倍和1.54倍,表现出更高FISH信号分辨率。检测到柚粗线期1号染色体着丝粒附近约3.01 Mb的探针信号缺失区。【结论】首次实现柑橘特定全长粗线期染色体的跟踪研究,为深入开展其结构和配对遗传利用的精准研究提供了新方法。展开更多
Sugarcane has recently attracted increasing attention for its potential as a source of sugar and bioethanol,so increasing its yield is essential to ensure the sugar security and bioenergy production.Intergeneric hybri...Sugarcane has recently attracted increasing attention for its potential as a source of sugar and bioethanol,so increasing its yield is essential to ensure the sugar security and bioenergy production.Intergeneric hybridization is a highly efficient method to produce new genetic variants of crop plants,particularly those species with high ploidy such as sugarcane(Saccharum spp.).Tripidium arundinaceum exhibits many desirable agronomic traits,and has been widely studied to produce hybrids with improved stress tolerance and other characteristics in sugarcane breeding.However,the genetic relationship between T.arundinaceum and Saccharum species,and the individual T.arundinaceum chromosomal compositions in sugarcane hybrids are still elusive.Here we used whole-genome single-nucleotide polymorphisms(SNPs)to ascertain the phylogenetic relationships between these species and found that T.arundinaceum is more closely related to Saccharum than Sorghum,in contrast to the previous narrow genetic analyses using chloroplast DNA.Additionally,oligonucleotide(oligo)-based chromosome-specific painting derived from Saccharum officinarum was able to distinctly identify the chromosomes of T.arundinaceum.We developed the oligo-genomic in situ hybridization(GISH)system for the first time,to unveil the novel chromosome translocations and the transmission of individual T.arundinaceum chromosomes in sugarcane progeny.Notably,we discovered that the chromosomal transmission of T.arundinaceum exhibited several different inheritance modes,including n,2n,and over 2n in the BC1 progenies.Such inheritance patterns may have resulted from first division restitution(FDR)or FDR+nondisjunction of a chromosome with the sister chromatids in the second meiosis division/second division restitution(FDR+NSC/SDR)model during meiosis.These results will be of substantial benefit for the further selection of T.arundinaceum chromosomes for sugarcane genetic improvement.展开更多
The wild decaploid species Thinopyrum ponticum(Podp.)Barkworth&D.R.Dewey is an important source of genes against biotic and abiotic stresses affecting wheat.The wheat–Th.ponticum partial amphiploid AUS6770 shows ...The wild decaploid species Thinopyrum ponticum(Podp.)Barkworth&D.R.Dewey is an important source of genes against biotic and abiotic stresses affecting wheat.The wheat–Th.ponticum partial amphiploid AUS6770 shows resistance to multiple diseases,including stripe rust,stem rust,and powdery mildew.Mitotic chromosomes of AUS6770 were characterized by non-denaturing-fluorescence in situ hybridization(ND-FISH),and the individual Th.ponticum chromosomes 1Ae to 7Ae were karyotypically distinguished by Oligo-FISH painting using bulked oligo pools based on wheat-barley collinear regions.A novel stripe rust resistant line A155,derived from AUS6770,was found to have 44 chromosomes,including a pair of 2Ae chromosomes and a pair of 6B-6Ae translocations.To detect plants with transfer of resistance genes from A155 to wheat chromosomes,1770 plants were developed from F2–F5 progenies of A155 crossed with the susceptible wheat cultivar MY11 and characterized with ND-FISH using multiple probes.A high frequency of transmission of chromosome 2Ae was observed,and 31 types of 2Ae chromosomal aberrations were identified using ND-FISH.Ten chromosomal bins on the 2Ae chromosome were determined from the deletion and translocation lines based on genome-based PCR markers.In combination with the evaluation of disease resistance,the gene(s)for stripe rust resistance was located on the FL0.79–1.00 of 2AeS and covers the corresponding region of 0–58.26 Mb in the reference genome of Th.elongatum.The newly identified wheat-Th.ponticum 2Ae translocation lines can be exploited as potential germplasm in wheat breeding for stripe rust resistance.展开更多
荧光原位杂交技术(fluorescence in situ hybridization,FISH)是植物分子细胞遗传学研究最为重要的手段之一。近些年,基于参考基因组设计的低拷贝寡聚核苷酸探针在FISH中应用得越来越广泛。然而,由于植物基因组中分布大量的重复序列,这...荧光原位杂交技术(fluorescence in situ hybridization,FISH)是植物分子细胞遗传学研究最为重要的手段之一。近些年,基于参考基因组设计的低拷贝寡聚核苷酸探针在FISH中应用得越来越广泛。然而,由于植物基因组中分布大量的重复序列,这使得oligo-FISH的分辨率存在一定局限性。利用包含多个荧光基团的荧光PCR引物,扩增出甘蔗染色体特异oligo探针,并进一步优化甘蔗的荧光原位杂交体系,提高了甘蔗oligo探针识别近缘物种染色体的效率。通过开发多荧光标记的甘蔗oligo探针以及甘蔗荧光杂交体系的优化,有效拓宽荧光信号的最小分辨率,提高信噪比(signal-to-noise ratio,SNR),并成功基于甘蔗oligo探针对高粱1-10号染色体分型。多荧光标记引物增强oligo探针信号的新方法及FISH体系的优化为今后在其他物种中提高oligo-FISH鉴定染色体及捕捉微弱的荧光信号提供了参考。展开更多
The Saccharum genus comprises species with large and variable chromosome numbers, leading to challenges in genomic studies and breeding improvement. Cytogenetics, including classical and molecular approaches, has play...The Saccharum genus comprises species with large and variable chromosome numbers, leading to challenges in genomic studies and breeding improvement. Cytogenetics, including classical and molecular approaches, has played a central role in deciphering the genome structure, classification, and evolution of the genus Saccharum. The application of fluorescence in situ hybridization using oligonucleotide probes significantly improved our understanding of the complex genomes of Saccharum species. This paper reviews the application and progress of cytogenetic techniques in Saccharum. Future applications of cytogenetics are discussed, as they could benefit both genomic studies and breeding of sugarcane as well as other plants with complex genomes.展开更多
单染色体识别为植物遗传学、生物多样性和进化研究的重要方向,也是大型基因组多倍体植物研究的难题之一。文章综述了单拷贝序列染色体涂染(single-copy sequence based chromosome painting)和寡核苷酸荧光原位杂交(oligonucleotide-FIS...单染色体识别为植物遗传学、生物多样性和进化研究的重要方向,也是大型基因组多倍体植物研究的难题之一。文章综述了单拷贝序列染色体涂染(single-copy sequence based chromosome painting)和寡核苷酸荧光原位杂交(oligonucleotide-FISH,Oligo-FISH)技术合成新型探针池的方法,总结其在比较基因组学研究中的应用案例,提出开发近缘物种通用寡核苷酸探针库的研究趋势,并对植物染色体分子核型技术在植物系统发育重建和植物育种中的应用进行了展望。展开更多
文摘【目的】深化柑橘减数分裂粗线期染色体结构组成和配对分析。【方法】首次利用柑橘染色体全长特异寡核苷酸荧光原位杂交(oligo-fluorescence in situ hybridization,oligo-FISH)涂染探针,精准示踪鉴定沙田柚粗线期1号染色体的结构特征,比较其与有丝分裂间期细胞核、有丝分裂前中期和中期染色体的结构差异,并揭示粗线期全长染色体的同源配对。【结果】观测到同源配对的全长粗线期与有丝分裂中期较大染色质结构差异。经测定,柚粗线期1号染色体平均全长、长臂长、短臂长和臂比分别为中期的13.93倍、16.54倍、10.44倍和1.54倍,表现出更高FISH信号分辨率。检测到柚粗线期1号染色体着丝粒附近约3.01 Mb的探针信号缺失区。【结论】首次实现柑橘特定全长粗线期染色体的跟踪研究,为深入开展其结构和配对遗传利用的精准研究提供了新方法。
基金funded by the Central Government and Local Science and Technology Development Special Project,China(2022L3086)the Sugarcane Research Foundation of Guangxi University,China(2022GZB006)+3 种基金supported by the National Natural Science Foundation of China(31771863)the Academy of Sugarcane and Sugar Industry,Guangxi University,China(ASSI-2023009)an independent fund of Guangxi Key Laboratory of Sugarcane Biology,China(GXKLSCB-20190201)the China Agriculture Research System of MOF and MARA(CARS-20-1-5)。
文摘Sugarcane has recently attracted increasing attention for its potential as a source of sugar and bioethanol,so increasing its yield is essential to ensure the sugar security and bioenergy production.Intergeneric hybridization is a highly efficient method to produce new genetic variants of crop plants,particularly those species with high ploidy such as sugarcane(Saccharum spp.).Tripidium arundinaceum exhibits many desirable agronomic traits,and has been widely studied to produce hybrids with improved stress tolerance and other characteristics in sugarcane breeding.However,the genetic relationship between T.arundinaceum and Saccharum species,and the individual T.arundinaceum chromosomal compositions in sugarcane hybrids are still elusive.Here we used whole-genome single-nucleotide polymorphisms(SNPs)to ascertain the phylogenetic relationships between these species and found that T.arundinaceum is more closely related to Saccharum than Sorghum,in contrast to the previous narrow genetic analyses using chloroplast DNA.Additionally,oligonucleotide(oligo)-based chromosome-specific painting derived from Saccharum officinarum was able to distinctly identify the chromosomes of T.arundinaceum.We developed the oligo-genomic in situ hybridization(GISH)system for the first time,to unveil the novel chromosome translocations and the transmission of individual T.arundinaceum chromosomes in sugarcane progeny.Notably,we discovered that the chromosomal transmission of T.arundinaceum exhibited several different inheritance modes,including n,2n,and over 2n in the BC1 progenies.Such inheritance patterns may have resulted from first division restitution(FDR)or FDR+nondisjunction of a chromosome with the sister chromatids in the second meiosis division/second division restitution(FDR+NSC/SDR)model during meiosis.These results will be of substantial benefit for the further selection of T.arundinaceum chromosomes for sugarcane genetic improvement.
基金supported by the National Natural Science Foundation of China (31971886)International Cooperation Program Program (2022YFH0012)of the Science and Technology Department of Sichuan.
文摘The wild decaploid species Thinopyrum ponticum(Podp.)Barkworth&D.R.Dewey is an important source of genes against biotic and abiotic stresses affecting wheat.The wheat–Th.ponticum partial amphiploid AUS6770 shows resistance to multiple diseases,including stripe rust,stem rust,and powdery mildew.Mitotic chromosomes of AUS6770 were characterized by non-denaturing-fluorescence in situ hybridization(ND-FISH),and the individual Th.ponticum chromosomes 1Ae to 7Ae were karyotypically distinguished by Oligo-FISH painting using bulked oligo pools based on wheat-barley collinear regions.A novel stripe rust resistant line A155,derived from AUS6770,was found to have 44 chromosomes,including a pair of 2Ae chromosomes and a pair of 6B-6Ae translocations.To detect plants with transfer of resistance genes from A155 to wheat chromosomes,1770 plants were developed from F2–F5 progenies of A155 crossed with the susceptible wheat cultivar MY11 and characterized with ND-FISH using multiple probes.A high frequency of transmission of chromosome 2Ae was observed,and 31 types of 2Ae chromosomal aberrations were identified using ND-FISH.Ten chromosomal bins on the 2Ae chromosome were determined from the deletion and translocation lines based on genome-based PCR markers.In combination with the evaluation of disease resistance,the gene(s)for stripe rust resistance was located on the FL0.79–1.00 of 2AeS and covers the corresponding region of 0–58.26 Mb in the reference genome of Th.elongatum.The newly identified wheat-Th.ponticum 2Ae translocation lines can be exploited as potential germplasm in wheat breeding for stripe rust resistance.
文摘荧光原位杂交技术(fluorescence in situ hybridization,FISH)是植物分子细胞遗传学研究最为重要的手段之一。近些年,基于参考基因组设计的低拷贝寡聚核苷酸探针在FISH中应用得越来越广泛。然而,由于植物基因组中分布大量的重复序列,这使得oligo-FISH的分辨率存在一定局限性。利用包含多个荧光基团的荧光PCR引物,扩增出甘蔗染色体特异oligo探针,并进一步优化甘蔗的荧光原位杂交体系,提高了甘蔗oligo探针识别近缘物种染色体的效率。通过开发多荧光标记的甘蔗oligo探针以及甘蔗荧光杂交体系的优化,有效拓宽荧光信号的最小分辨率,提高信噪比(signal-to-noise ratio,SNR),并成功基于甘蔗oligo探针对高粱1-10号染色体分型。多荧光标记引物增强oligo探针信号的新方法及FISH体系的优化为今后在其他物种中提高oligo-FISH鉴定染色体及捕捉微弱的荧光信号提供了参考。
基金supported by the Startup Foundation from Nantong University (03083074)the National Natural Science Foundation of China (32070544)。
文摘The Saccharum genus comprises species with large and variable chromosome numbers, leading to challenges in genomic studies and breeding improvement. Cytogenetics, including classical and molecular approaches, has played a central role in deciphering the genome structure, classification, and evolution of the genus Saccharum. The application of fluorescence in situ hybridization using oligonucleotide probes significantly improved our understanding of the complex genomes of Saccharum species. This paper reviews the application and progress of cytogenetic techniques in Saccharum. Future applications of cytogenetics are discussed, as they could benefit both genomic studies and breeding of sugarcane as well as other plants with complex genomes.
文摘单染色体识别为植物遗传学、生物多样性和进化研究的重要方向,也是大型基因组多倍体植物研究的难题之一。文章综述了单拷贝序列染色体涂染(single-copy sequence based chromosome painting)和寡核苷酸荧光原位杂交(oligonucleotide-FISH,Oligo-FISH)技术合成新型探针池的方法,总结其在比较基因组学研究中的应用案例,提出开发近缘物种通用寡核苷酸探针库的研究趋势,并对植物染色体分子核型技术在植物系统发育重建和植物育种中的应用进行了展望。
