【目的】利用大数据比较2条不同的簇毛麦6V(#2和#4)染色体及其与小麦6A、6D染色体间DNA水平上的差异,为小麦-簇毛麦靶向易位的精准设计育种提供依据。【方法】以6V#4(6D)异代换系RW15为父本和6V#2(6A)异代换系南87-88为母本进行杂交,获...【目的】利用大数据比较2条不同的簇毛麦6V(#2和#4)染色体及其与小麦6A、6D染色体间DNA水平上的差异,为小麦-簇毛麦靶向易位的精准设计育种提供依据。【方法】以6V#4(6D)异代换系RW15为父本和6V#2(6A)异代换系南87-88为母本进行杂交,获得F2分离群体,利用6V#4S/6V#2S/6AS/6DS/6VL特异分子标记检测F2植株,筛选新类型的代换系,并用分子标记结合基因组原位杂交(genomic in situ hybridization,GISH)对新类型代换系进行确认,再利用小麦55K芯片中的6A、6D探针,对新代换系及其双亲南87-88和RW15进行分析;结合660K芯片6A、6D探针对2份簇毛麦的SNP分析结果,筛选6V特异SNP。【结果】GISH分析表明,19EL124和19EL134的体细胞染色体数2n=42,分别携带2条完整的外源染色体;分子标记鉴定结果表明,19EL124含有6V#4S/6DS特异标记带,缺失了6V#2S/6AS特异带,而19EL134含有6V#2S/6AS特异标记带,缺失了6V#4S/6DS特异带;19EL124和19EL134都含有6VL的特异带,证明19EL124为6V#4(6A)异代换系,19EL134为6V#2(6D)异代换系。55K芯片检测结果表明,异代换系中关键染色体探针的检测效率显著低于其他染色体,且对同类型异代换不同系的检测效率也有所不同。1177个6A探针中,63.21%不能对6A代换系南87-88分型,68.90%不能对6A异代换系19EL124分型,22.51%检测到6V#2和6V#4间的多态性,其中88个只能检测到6V#4染色体,而155个只能检测到6V#2染色体;479个6D探针中,49.48%不能对6D异代换系RW15分型,53.44%不能对6D代换系19EL134分型,16.70%检测到6V#2和6V#4间的多态性,其中23个只能检测到6V#2染色体,42个只能检测到6V#4染色体。整合55K和660K芯片的共有探针,分别从395个6A、231个6D探针筛选获得簇毛麦6V特异的SNP标记22个和15个,其中3个可在6V#2和6V#4染色体间显示多态性。【结论】小麦染色体的缺失与外源染色体的替换,使相应染色体探针的检测效率大幅降低,NA分型比例极大增加,且多数NA分型在2条不同的外源染色体间显示多态;相同探针对2条外源染色体的检测效率不同,小麦6A探针可以更好地检测6V#2,而小麦6D探针可更好检测6V#4;在簇毛麦与异代换系6V染色体的一致性分型中,筛选获得簇毛麦6V特异的SNP标记37个。展开更多
Under stress conditions such as droughthigh-salinity and low-temperature, the transcription factorof DREB (dehydration responsive element binding proteins)improved efficiently stress resistance by regulating the ex-pr...Under stress conditions such as droughthigh-salinity and low-temperature, the transcription factorof DREB (dehydration responsive element binding proteins)improved efficiently stress resistance by regulating the ex-pression of its downstream genes with various environmentastress resistance in plants. GmDREB gene (GenBank Acces-sion No. AF514908) encoding a stress-inducible transcriptionfactor was cloned by screening a cDNA library of Glycinemax cv. Jinong 27 with yeast one-hybrid method. GmDREBgene was 910 bp in length and encoded 174 amino acids con-taining a conserved AP2/EREBP DNA-binding domain of 58amino acids. Two conserved functional amino acids, valineand glutamic acid, were located on the 14th and the 19thamino acid residues in the conserved structural domain. Analkaline amino acid region (KKR) related to a nuclear local-ization signal was at the N-terminal, while an acidic aminoacid region (DDD) related to trans-activation was at theC-terminal. Plant expression vectors were constructed andtransformed into wheat by bombardment. In total, 13 trans-genic plants with Ubi::GmDREB and 11 transgenic plantswith rd29A::GmDREB were identified from 103 regenerationplants by molecular analysis. The drought and salt tolerancesof T1 transgenic lines with Ubi::GmDREB orrd29A::GmDREB were demonstrated to be improved ascompared to wild type. The result also suggested that bothUbiquitin and rd29A promoters could effectively drive theexpression of the GmDREB gene and enhance drought andsalt tolerance of T1 plants.展开更多
文摘【目的】利用大数据比较2条不同的簇毛麦6V(#2和#4)染色体及其与小麦6A、6D染色体间DNA水平上的差异,为小麦-簇毛麦靶向易位的精准设计育种提供依据。【方法】以6V#4(6D)异代换系RW15为父本和6V#2(6A)异代换系南87-88为母本进行杂交,获得F2分离群体,利用6V#4S/6V#2S/6AS/6DS/6VL特异分子标记检测F2植株,筛选新类型的代换系,并用分子标记结合基因组原位杂交(genomic in situ hybridization,GISH)对新类型代换系进行确认,再利用小麦55K芯片中的6A、6D探针,对新代换系及其双亲南87-88和RW15进行分析;结合660K芯片6A、6D探针对2份簇毛麦的SNP分析结果,筛选6V特异SNP。【结果】GISH分析表明,19EL124和19EL134的体细胞染色体数2n=42,分别携带2条完整的外源染色体;分子标记鉴定结果表明,19EL124含有6V#4S/6DS特异标记带,缺失了6V#2S/6AS特异带,而19EL134含有6V#2S/6AS特异标记带,缺失了6V#4S/6DS特异带;19EL124和19EL134都含有6VL的特异带,证明19EL124为6V#4(6A)异代换系,19EL134为6V#2(6D)异代换系。55K芯片检测结果表明,异代换系中关键染色体探针的检测效率显著低于其他染色体,且对同类型异代换不同系的检测效率也有所不同。1177个6A探针中,63.21%不能对6A代换系南87-88分型,68.90%不能对6A异代换系19EL124分型,22.51%检测到6V#2和6V#4间的多态性,其中88个只能检测到6V#4染色体,而155个只能检测到6V#2染色体;479个6D探针中,49.48%不能对6D异代换系RW15分型,53.44%不能对6D代换系19EL134分型,16.70%检测到6V#2和6V#4间的多态性,其中23个只能检测到6V#2染色体,42个只能检测到6V#4染色体。整合55K和660K芯片的共有探针,分别从395个6A、231个6D探针筛选获得簇毛麦6V特异的SNP标记22个和15个,其中3个可在6V#2和6V#4染色体间显示多态性。【结论】小麦染色体的缺失与外源染色体的替换,使相应染色体探针的检测效率大幅降低,NA分型比例极大增加,且多数NA分型在2条不同的外源染色体间显示多态;相同探针对2条外源染色体的检测效率不同,小麦6A探针可以更好地检测6V#2,而小麦6D探针可更好检测6V#4;在簇毛麦与异代换系6V染色体的一致性分型中,筛选获得簇毛麦6V特异的SNP标记37个。
基金This work was supported by the National 863 Project(Grant No.2002AA224081)National Special Project for Plant Transgenic and Industry(Grant No.JY03-A-18).
文摘Under stress conditions such as droughthigh-salinity and low-temperature, the transcription factorof DREB (dehydration responsive element binding proteins)improved efficiently stress resistance by regulating the ex-pression of its downstream genes with various environmentastress resistance in plants. GmDREB gene (GenBank Acces-sion No. AF514908) encoding a stress-inducible transcriptionfactor was cloned by screening a cDNA library of Glycinemax cv. Jinong 27 with yeast one-hybrid method. GmDREBgene was 910 bp in length and encoded 174 amino acids con-taining a conserved AP2/EREBP DNA-binding domain of 58amino acids. Two conserved functional amino acids, valineand glutamic acid, were located on the 14th and the 19thamino acid residues in the conserved structural domain. Analkaline amino acid region (KKR) related to a nuclear local-ization signal was at the N-terminal, while an acidic aminoacid region (DDD) related to trans-activation was at theC-terminal. Plant expression vectors were constructed andtransformed into wheat by bombardment. In total, 13 trans-genic plants with Ubi::GmDREB and 11 transgenic plantswith rd29A::GmDREB were identified from 103 regenerationplants by molecular analysis. The drought and salt tolerancesof T1 transgenic lines with Ubi::GmDREB orrd29A::GmDREB were demonstrated to be improved ascompared to wild type. The result also suggested that bothUbiquitin and rd29A promoters could effectively drive theexpression of the GmDREB gene and enhance drought andsalt tolerance of T1 plants.