将小麦近缘属植物黑麦中的优良基因导入小麦可以拓宽小麦的遗传基础,丰富小麦的遗传变异。本研究调查并分析了390份小麦-黑麦种质材料。在这390份种质材料中,6个主要农艺性状值均有较大的极差,说明其遗传多样性丰富。与10份小麦主栽品...将小麦近缘属植物黑麦中的优良基因导入小麦可以拓宽小麦的遗传基础,丰富小麦的遗传变异。本研究调查并分析了390份小麦-黑麦种质材料。在这390份种质材料中,6个主要农艺性状值均有较大的极差,说明其遗传多样性丰富。与10份小麦主栽品种相比,90%以上的材料具有穗长和分蘖数的显著优势,60%以上的材料具有小穗数优势,约30%的材料穗粒数和千粒重显著高于主栽品种。利用基因组原位杂交(genomic in situ hybridization,GISH)和多色荧光原位杂交(multicolor fluorescent in situ hybridization,mc-FISH)技术,对8份农艺性状优良的代表性材料进行染色体组成分析,发现3份为六倍体小黑麦(AABBRR),2份为八倍体小黑麦(AABBDDRR),1份为1RS?1BL易位系,其余2份不具有可见的黑麦染色体或染色体片段。值得指出的是,3份六倍体小黑麦与2份八倍体小黑麦所含的黑麦染色体不完全相同。八倍体小黑麦中有1对来源于黑麦的小染色体,而六倍体小黑麦中没有类似小染色体;并且,不同材料中黑麦4R染色体端部的GISH杂交带有明显差异。本研究结果为这些小麦-黑麦种质材料进一步应用于小麦育种提供了依据。展开更多
【目的】针对花生染色体较小,染色体细胞学标记少,细胞遗传研究相对滞后,染色体分类识别困难的问题,建立能够准确区分栽培花生(Arachis hypogaea L.,2n=4x=40,AABB)A、B染色体组的新核型,提高染色体识别准确率,以揭示栽培花生和野生供...【目的】针对花生染色体较小,染色体细胞学标记少,细胞遗传研究相对滞后,染色体分类识别困难的问题,建立能够准确区分栽培花生(Arachis hypogaea L.,2n=4x=40,AABB)A、B染色体组的新核型,提高染色体识别准确率,以揭示栽培花生和野生供体亲本的染色体对应关系,鉴定栽培种花生染色体结构变异体。【方法】以花生栽培种(Arachis hypogaea L.,2n=4x=40,AABB)的2个可能供体亲本即花生野生种Arachis duranensis(2n=2x=20,BB)和Arachis ipa?nsis(2n=2x=20,AA)全基因组DNA及5S r DNA和45S r DNA为探针,利用顺序基因组荧光原位杂交(GISH)和多色荧光原位杂交(Mc FISH)技术(简称顺序GISH-FISH)结合DAPI染色,在准确区分花生栽培种A、B染色体组的基础上,对花生栽培品种Z5163及其供体亲本染色体进行分析,建立花生栽培种新核型,并利用该核型对其他栽培品种的染色体进行分析,以探讨该核型的应用潜力和栽培花生染色体组成特点。【结果】以A.ipa?nsis和A.duranensis全基因组DNA为探针的GISH分析表明,以A.ipa?nsis为探针在花生栽培种20条B组染色体上能够产生清晰稳定的杂交信号,在A组染色体上没有信号,而以A.duranensis为探针,只在18条A组染色体能产生信号,但1对A组的小染色体"A染色体"不易被区分,因此,以A.ipa?nsis为探针可以准确区分花生栽培种A、B染色体组;综合5S r DNA和45S r DNA Mc-FISH和DAPI染色分析,发现花生栽培种A、B染色体组DAPI带纹、5S r DNA和45S r DNA的分布分别与A.duranensis和A.ipa?nsis一致,此结果支持A.duranensis和A.ipa?nsis是花生栽培种的供体亲本。DAPI染色结果显示,A.ipa?nsis及花生栽培种的B组染色体均有14条染色体显示着丝粒带纹,明显多于前人报道,表明仅利用DAPI染色来区分花生栽培种A、B组染色体的方法具有局限性。综合DAPI染色、r DNA、A.duranensis和A.ipa?nsis基因组探针进行顺序GISH-FISH分析,建立了可以准确识别花生栽培种A、B染色体组新核型。然后利用该核型对3个栽培种品种的染色体组成进行了分析,首次发现一个自发的花生染色体代换系MS B1(A1),揭示了栽培花生染色体B1与A1之间存在部分同源关系。【结论】野生花生A.duranensis和A.ipa?nsis分别与栽培花生A和B基因组染色体间具有很好的对应关系;研究建立的基于GISH-FISH和DAPI染色的栽培花生新核型,不但可以准确区分大部分A、B组染色体,而且还能识别栽培花生在多倍体化和人工进化过程中可能存在的自发的染色体变异,揭示A、B组染色体间的部分同源性。展开更多
Thinopyrum intermedium(2 n = 6 x = 42, JJJ^sJ^sStSt) has been hybridized extensively with common wheat and has proven to be a valuable germplasm source for improving disease resistance, quality attributes, and yield p...Thinopyrum intermedium(2 n = 6 x = 42, JJJ^sJ^sStSt) has been hybridized extensively with common wheat and has proven to be a valuable germplasm source for improving disease resistance, quality attributes, and yield potential in wheat. We characterized new disease resistant wheat-Th. intermedium derivatives A1082 and A5-5 using sequential multi-color fluorescence in situ hybridization(mc-FISH), genomic in situ hybridization(GISH), PCRbased landmark unique gene(PLUG) and intron targeting(IT) markers. A1082 was identified as a wheat-Th. intermedium 3J disomic addition line, and A5-5 was a T4BS·5J^sL homozygous Robertsonian translocation line. Seventy-one and 106 pairs of primers amplified Th.intermedium-specific bands allowing chromosomes 3J and 5J^s to be tracked, respectively. A new oligonucleotide probe, Oligo-6 H-2-100, was developed for FISH labeling of the subterminal region of the long arm of chromosome 5J^s. Both lines were highly resistant to stripe rust pathogen races prevalent in Chinese field screening nurseries. A5-5 also displayed a significant increase in tiller number compared to its wheat parent. The new lines can be exploited as useful germplasms for wheat improvement.展开更多
Hordeum californicum （2n = 2x = 14, HH） is resistant to several wheat diseases and tolerant to lower nitrogen. In this study, a molecular karyotype of H. californicum chromosomes in the Triticum aestivum L. cv. Chin...Hordeum californicum （2n = 2x = 14, HH） is resistant to several wheat diseases and tolerant to lower nitrogen. In this study, a molecular karyotype of H. californicum chromosomes in the Triticum aestivum L. cv. Chinese Spring （CS）-H. californicum amphidiploid （2n = 6x = 56, AABBDDHH） was established. By genomic in situ hybridization （GISH） and multicolor fluorescent in situ hybridization （FISH） using repetitive DNA clones （pTa71, pTa794 and pSc119.2） as probes, the H. californicum chromosomes could be differentiated from each other and from the wheat chromosomes unequivocally. Based on molecular karyotype and marker analyses, 12 wheat--alien chromosome lines, including four disomic addition lines （DAH1, DAH3, DAH5 and DAH6）, five telosomic addition lines （MtH7L, MtHIS, MtH1L, DtH6S and DtH6L）, one multiple addition line involving H. californicum chromosome H2, one disomic substitution line （DSH4） and one translocation line （TH7S/1BL）, were identified from the progenies derived from the crosses of CS-H. californicum amphidiploid with common wheat varieties. A total of 482 EST （expressed sequence tag） or SSR （simple sequence repeat） markers specific for individual H. californicum chromosomes were identified, and 47, 50, 45, 49, 21, 51 and 40 markers were assigned to chromosomes H1, H2, H3, H4, H5, H6 and H7, respectively. According to the chromosome allocation of these markers, chromosomes H2, H3, H4, H5, and H7 of H. californicum have relationship with wheat homoeologous groups 5, 2, 6, 3, and 1, and hence could be designated as 5Hc, 2He, 6Hc, 3Hc and 1Hc, respectively. The chromosomes H1 and H6 were designated as 7Hc and 4Hc, respectively, by referring to SSR markers located on rye chromosomes.展开更多
Rye(Secale cereale)is a valuable gene donor for wheat improvement,especially for its resistance to diseases.Developing rye-derived resistance sources is important for wheat breeding.In the present study,two wheat-rye ...Rye(Secale cereale)is a valuable gene donor for wheat improvement,especially for its resistance to diseases.Developing rye-derived resistance sources is important for wheat breeding.In the present study,two wheat-rye derivatives,designated JS016 and JS110,were produced by crossing common wheat cultivar Yangmai 23 with Pakistani rye accession W2A.Using sequential genomic in situ hybridization(GISH)and multicolor fluorescence in situ hybridization(mc-FISH),JS016 and JS110 were identified as a T6BS.6RL translocation line and a T6BS.6BL6RL translocation line,respectively.Ten newly 6RL chromosome arm-specific markers were developed and used to confirm the 6RL translocation.The wheat 55K single-nucleotide polymorphism(SNP)array further verified the molecular cytogenetic identification results above and clarified their breakpoints at 430.9 and 523.0 Mb of chromosome 6B in JS016 and JS110,respectively.Resistance spectrum and allelism test demonstrated that JS016 and JS110 possessed novel powdery mildew resistance gene(s)that was derived from the 6RL translocation but differed from Pm20.Moreover,JS016 and JS110 had better agronomic traits than the previously reported 6RL translocation line carrying Pm20.To efficiently transfer and detect the 6RL translocation from JS016 and JS110,one 6RL-specific Kompetitive allele specific PCR(KASP)marker was developed and validated in high throughput marker-assisted selection(MAS).展开更多
文摘将小麦近缘属植物黑麦中的优良基因导入小麦可以拓宽小麦的遗传基础,丰富小麦的遗传变异。本研究调查并分析了390份小麦-黑麦种质材料。在这390份种质材料中,6个主要农艺性状值均有较大的极差,说明其遗传多样性丰富。与10份小麦主栽品种相比,90%以上的材料具有穗长和分蘖数的显著优势,60%以上的材料具有小穗数优势,约30%的材料穗粒数和千粒重显著高于主栽品种。利用基因组原位杂交(genomic in situ hybridization,GISH)和多色荧光原位杂交(multicolor fluorescent in situ hybridization,mc-FISH)技术,对8份农艺性状优良的代表性材料进行染色体组成分析,发现3份为六倍体小黑麦(AABBRR),2份为八倍体小黑麦(AABBDDRR),1份为1RS?1BL易位系,其余2份不具有可见的黑麦染色体或染色体片段。值得指出的是,3份六倍体小黑麦与2份八倍体小黑麦所含的黑麦染色体不完全相同。八倍体小黑麦中有1对来源于黑麦的小染色体,而六倍体小黑麦中没有类似小染色体;并且,不同材料中黑麦4R染色体端部的GISH杂交带有明显差异。本研究结果为这些小麦-黑麦种质材料进一步应用于小麦育种提供了依据。
文摘【目的】针对花生染色体较小,染色体细胞学标记少,细胞遗传研究相对滞后,染色体分类识别困难的问题,建立能够准确区分栽培花生(Arachis hypogaea L.,2n=4x=40,AABB)A、B染色体组的新核型,提高染色体识别准确率,以揭示栽培花生和野生供体亲本的染色体对应关系,鉴定栽培种花生染色体结构变异体。【方法】以花生栽培种(Arachis hypogaea L.,2n=4x=40,AABB)的2个可能供体亲本即花生野生种Arachis duranensis(2n=2x=20,BB)和Arachis ipa?nsis(2n=2x=20,AA)全基因组DNA及5S r DNA和45S r DNA为探针,利用顺序基因组荧光原位杂交(GISH)和多色荧光原位杂交(Mc FISH)技术(简称顺序GISH-FISH)结合DAPI染色,在准确区分花生栽培种A、B染色体组的基础上,对花生栽培品种Z5163及其供体亲本染色体进行分析,建立花生栽培种新核型,并利用该核型对其他栽培品种的染色体进行分析,以探讨该核型的应用潜力和栽培花生染色体组成特点。【结果】以A.ipa?nsis和A.duranensis全基因组DNA为探针的GISH分析表明,以A.ipa?nsis为探针在花生栽培种20条B组染色体上能够产生清晰稳定的杂交信号,在A组染色体上没有信号,而以A.duranensis为探针,只在18条A组染色体能产生信号,但1对A组的小染色体"A染色体"不易被区分,因此,以A.ipa?nsis为探针可以准确区分花生栽培种A、B染色体组;综合5S r DNA和45S r DNA Mc-FISH和DAPI染色分析,发现花生栽培种A、B染色体组DAPI带纹、5S r DNA和45S r DNA的分布分别与A.duranensis和A.ipa?nsis一致,此结果支持A.duranensis和A.ipa?nsis是花生栽培种的供体亲本。DAPI染色结果显示,A.ipa?nsis及花生栽培种的B组染色体均有14条染色体显示着丝粒带纹,明显多于前人报道,表明仅利用DAPI染色来区分花生栽培种A、B组染色体的方法具有局限性。综合DAPI染色、r DNA、A.duranensis和A.ipa?nsis基因组探针进行顺序GISH-FISH分析,建立了可以准确识别花生栽培种A、B染色体组新核型。然后利用该核型对3个栽培种品种的染色体组成进行了分析,首次发现一个自发的花生染色体代换系MS B1(A1),揭示了栽培花生染色体B1与A1之间存在部分同源关系。【结论】野生花生A.duranensis和A.ipa?nsis分别与栽培花生A和B基因组染色体间具有很好的对应关系;研究建立的基于GISH-FISH和DAPI染色的栽培花生新核型,不但可以准确区分大部分A、B组染色体,而且还能识别栽培花生在多倍体化和人工进化过程中可能存在的自发的染色体变异,揭示A、B组染色体间的部分同源性。
基金funded by the National Key Research and Development Program of China (2016YFD0102000)Applied and Basic Project (2016JY0075) from Science and Technology Department of Sichuan Province, ChinaNational Natural Science Foundation of China (No. 31171542)
文摘Thinopyrum intermedium(2 n = 6 x = 42, JJJ^sJ^sStSt) has been hybridized extensively with common wheat and has proven to be a valuable germplasm source for improving disease resistance, quality attributes, and yield potential in wheat. We characterized new disease resistant wheat-Th. intermedium derivatives A1082 and A5-5 using sequential multi-color fluorescence in situ hybridization(mc-FISH), genomic in situ hybridization(GISH), PCRbased landmark unique gene(PLUG) and intron targeting(IT) markers. A1082 was identified as a wheat-Th. intermedium 3J disomic addition line, and A5-5 was a T4BS·5J^sL homozygous Robertsonian translocation line. Seventy-one and 106 pairs of primers amplified Th.intermedium-specific bands allowing chromosomes 3J and 5J^s to be tracked, respectively. A new oligonucleotide probe, Oligo-6 H-2-100, was developed for FISH labeling of the subterminal region of the long arm of chromosome 5J^s. Both lines were highly resistant to stripe rust pathogen races prevalent in Chinese field screening nurseries. A5-5 also displayed a significant increase in tiller number compared to its wheat parent. The new lines can be exploited as useful germplasms for wheat improvement.
基金supported by the Technology Support Program of Jiangsu Province (No. BE2012306)the Program of Introducing Talents of Discipline to Universities (No. B08025)+1 种基金the Project No.7 from Science and High-Tech Based Major Program of Agriculture Committee of Shanghai Municipal Administration (No.20127)the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)
文摘Hordeum californicum （2n = 2x = 14, HH） is resistant to several wheat diseases and tolerant to lower nitrogen. In this study, a molecular karyotype of H. californicum chromosomes in the Triticum aestivum L. cv. Chinese Spring （CS）-H. californicum amphidiploid （2n = 6x = 56, AABBDDHH） was established. By genomic in situ hybridization （GISH） and multicolor fluorescent in situ hybridization （FISH） using repetitive DNA clones （pTa71, pTa794 and pSc119.2） as probes, the H. californicum chromosomes could be differentiated from each other and from the wheat chromosomes unequivocally. Based on molecular karyotype and marker analyses, 12 wheat--alien chromosome lines, including four disomic addition lines （DAH1, DAH3, DAH5 and DAH6）, five telosomic addition lines （MtH7L, MtHIS, MtH1L, DtH6S and DtH6L）, one multiple addition line involving H. californicum chromosome H2, one disomic substitution line （DSH4） and one translocation line （TH7S/1BL）, were identified from the progenies derived from the crosses of CS-H. californicum amphidiploid with common wheat varieties. A total of 482 EST （expressed sequence tag） or SSR （simple sequence repeat） markers specific for individual H. californicum chromosomes were identified, and 47, 50, 45, 49, 21, 51 and 40 markers were assigned to chromosomes H1, H2, H3, H4, H5, H6 and H7, respectively. According to the chromosome allocation of these markers, chromosomes H2, H3, H4, H5, and H7 of H. californicum have relationship with wheat homoeologous groups 5, 2, 6, 3, and 1, and hence could be designated as 5Hc, 2He, 6Hc, 3Hc and 1Hc, respectively. The chromosomes H1 and H6 were designated as 7Hc and 4Hc, respectively, by referring to SSR markers located on rye chromosomes.
基金supported by the National Natural Science Foundation of China(32171990 and 32072053)Key Research and Development Program of Zhenjiang(NY2021001)+4 种基金State Key Laboratory of Plant Cell and Chromosome Engineering(PCCE-KF-2021-05 and PCCE-KF-2022-07)State Key Laboratory of Crop Biology in Shandong Agricultural University(2021KF01)Natural Science Foundation of the Jiangsu Higher Education institutions of China(21KJB210004)Open Project Funding of State Key Laboratory of Crop Stress Adaptation and Improvement(CX1130A0920014)Key Research and Development Program of Shandong Province(2020CXGC010805).
文摘Rye(Secale cereale)is a valuable gene donor for wheat improvement,especially for its resistance to diseases.Developing rye-derived resistance sources is important for wheat breeding.In the present study,two wheat-rye derivatives,designated JS016 and JS110,were produced by crossing common wheat cultivar Yangmai 23 with Pakistani rye accession W2A.Using sequential genomic in situ hybridization(GISH)and multicolor fluorescence in situ hybridization(mc-FISH),JS016 and JS110 were identified as a T6BS.6RL translocation line and a T6BS.6BL6RL translocation line,respectively.Ten newly 6RL chromosome arm-specific markers were developed and used to confirm the 6RL translocation.The wheat 55K single-nucleotide polymorphism(SNP)array further verified the molecular cytogenetic identification results above and clarified their breakpoints at 430.9 and 523.0 Mb of chromosome 6B in JS016 and JS110,respectively.Resistance spectrum and allelism test demonstrated that JS016 and JS110 possessed novel powdery mildew resistance gene(s)that was derived from the 6RL translocation but differed from Pm20.Moreover,JS016 and JS110 had better agronomic traits than the previously reported 6RL translocation line carrying Pm20.To efficiently transfer and detect the 6RL translocation from JS016 and JS110,one 6RL-specific Kompetitive allele specific PCR(KASP)marker was developed and validated in high throughput marker-assisted selection(MAS).
