To break the narrow diversity bottleneck of the wheat D genome, a set of Aegilops tauschii-wheat introgression(A-WI) lines was developed by crossing Ae. tauschii accession T015 with common wheat elite cultivar Zhoumai...To break the narrow diversity bottleneck of the wheat D genome, a set of Aegilops tauschii-wheat introgression(A-WI) lines was developed by crossing Ae. tauschii accession T015 with common wheat elite cultivar Zhoumai 18(Zhou18). A high-density genetic map was constructed based on Single Nucleotide Polymorphism(SNP) markers and 15 yield-related traits were evaluated in 11 environments for detecting quantitative trait loci(QTL). A total of 27 environmentally stable QTL were identified in at least five environments, 20 of which were derived from Ae. tauschii T015, explaining up to 24.27% of the phenotypic variations. The major QTL for kernel length(KL), QKl-2D.5, was delimited to a physical interval of approximately 2.6 Mb harboring 52 candidate genes. Three Kompetitive Allele Specific PCR(KASP)markers were successfully developed based on nonsynonymous nucleotide mutations of candidate gene AetT093_2Dv1G100900.1 and showed that A-WI lines with the T015 haplotype had significantly longer KL than the Zhou18 haplotype across all 11 environments. Four primary valuable A-WIs with good trait performance and carrying yield-related QTL were selected for breeding improvement. The results will facilitate the efficient transfer of beneficial genes from Ae. tauschii into wheat cultivars to improve wheat yield and other traits.展开更多
A total of 43 unique clones(Z4A-1 to Z4A-43 with GenBank accession numbers of HM120221, HM120222, JX828270, JN831402 to JN831406, and KC715889 to KC715923, respectively) were amplified and cloned from common wheat cul...A total of 43 unique clones(Z4A-1 to Z4A-43 with GenBank accession numbers of HM120221, HM120222, JX828270, JN831402 to JN831406, and KC715889 to KC715923, respectively) were amplified and cloned from common wheat cultivar Zhengmai 004 using a PCR-based strategy. They included 22 full-ORF α-gliadin genes and 21 pseudogenes containing at least one in-frame stop codon. Comparative analysis of the deduced amino acid sequences showed that all the isolated genes displayed the typical structural features of α-gliadin genes and that the putative proteins of Z4A-7, Z4A-14, Z4A-17 and Z4A-20 had an extra cysteine residue in the unique domain II, while Z4A-15 lacked the second conserved cysteine residue in the unique domain I. The two fusion proteins of Z4A-15 and Z4A-20 were successfully detected by SDS-PAGE and Western blotting, although the protein level was relatively low. Based on the occurrence of the four major epitopes, as well as the lengths of the two glutamine repeats, 8, 6, and 8 genes were assigned to the Gli-2 loci on the respective chromosomes 6A, 6B, and 6D and a total of respectively 16, 0 and 23 immunogenic peptides were identified. In addition, 4 of the 5 genes with odd numbers of cysteine residues were assigned to chromosome 6D, suggesting that common wheat cultivar Zhengmai 004 has the potential to induce celiac disease(CD) and that the D genome exerts the most influence on gluten quality, but is the most deleterious for CD patients. By phylogenetic analysis, 11 exceptional α-gliadins with few or no immunogenic peptides from Triticum monococcum and Aegilops tauschii were detected, a finding that further supports the prospect of CD prevention. Finally, secondary structure prediction showed that most(98.48%) of the α-gliadins invariably contained five conserved α-helices(H1 to H5) in the two glutamine repeats and unique domains and 67.68% of the α-gliadins also contained a β-strand(S) in the C-terminal unique domains. An absent α-helix H2, 1–2 extra α-helices, or an additional β-strand(SE) also probably occurred in some cases. Of the 22 cloned genes in this work, 10 putative proteins contained 1–2 extra α-helices in addition to the five conserved α-helices or the additional β-strand. The observation that most of the α-helices and β-strands were present in the two unique domains and that an extra α-helix also probably occurred in the two glutamine repeats in some desirable genes strongly suggested that these two uniquedomains are the most important regions for the function of α-gliadins, although the glutamine repeats would also contribute in some cases.展开更多
Horizontal gene transfer(HGT) refers to the flow of genetic materials to non-offspring,and occasionally HGT in plants can improve the adaptation of organisms in new niches due to expanded metabolic capability.Anthocya...Horizontal gene transfer(HGT) refers to the flow of genetic materials to non-offspring,and occasionally HGT in plants can improve the adaptation of organisms in new niches due to expanded metabolic capability.Anthocyanins are an important group of water-soluble red,purple,or blue secondary metabolites,whose diversity results from modification after the main skeleton biosynthesis.Cuscuta is a stem holoparasitic genus,whose members form direct connection with hosts to withdraw water,nutrients,and macromolecules.Such intimate association is thought to increase the frequency of HGT.By transcriptome screening for foreign genes in Cuscuta australis,we discovered that one gene encoding a putative anthocyanin acyltransferase gene of the BAHD family,which is likely to be involved in anthocyanin modification,was acquired by C.australis from Fabaceae through HGT.The anthocyanin acyltransferase-like(AT-like) gene was confirmed to be present in the genome assembly of C.australis and the transcriptomes of Cuscuta pentagona.The higher transcriptional level in old stems is consistent with its putative function in secondary metabolism by stabilizing anthocyanin at neutral pH and thus HGT of this AT-like gene may have improved biotic and abiotic resistance of Cuscuta.展开更多
基金financially supported by the National Natural Science Foundation of China (32230079, 32001492, 31871615, and31901547)Natural Science Foundation of Henan Province(222301420102)。
文摘To break the narrow diversity bottleneck of the wheat D genome, a set of Aegilops tauschii-wheat introgression(A-WI) lines was developed by crossing Ae. tauschii accession T015 with common wheat elite cultivar Zhoumai 18(Zhou18). A high-density genetic map was constructed based on Single Nucleotide Polymorphism(SNP) markers and 15 yield-related traits were evaluated in 11 environments for detecting quantitative trait loci(QTL). A total of 27 environmentally stable QTL were identified in at least five environments, 20 of which were derived from Ae. tauschii T015, explaining up to 24.27% of the phenotypic variations. The major QTL for kernel length(KL), QKl-2D.5, was delimited to a physical interval of approximately 2.6 Mb harboring 52 candidate genes. Three Kompetitive Allele Specific PCR(KASP)markers were successfully developed based on nonsynonymous nucleotide mutations of candidate gene AetT093_2Dv1G100900.1 and showed that A-WI lines with the T015 haplotype had significantly longer KL than the Zhou18 haplotype across all 11 environments. Four primary valuable A-WIs with good trait performance and carrying yield-related QTL were selected for breeding improvement. The results will facilitate the efficient transfer of beneficial genes from Ae. tauschii into wheat cultivars to improve wheat yield and other traits.
基金supported by the National Natural Science Foundation of China (31271713)the "Twelfth Five-Year-Plan" in National Science and Technology for Rural Development in China (2011BAD07B01 and 2012AA101105)
文摘A total of 43 unique clones(Z4A-1 to Z4A-43 with GenBank accession numbers of HM120221, HM120222, JX828270, JN831402 to JN831406, and KC715889 to KC715923, respectively) were amplified and cloned from common wheat cultivar Zhengmai 004 using a PCR-based strategy. They included 22 full-ORF α-gliadin genes and 21 pseudogenes containing at least one in-frame stop codon. Comparative analysis of the deduced amino acid sequences showed that all the isolated genes displayed the typical structural features of α-gliadin genes and that the putative proteins of Z4A-7, Z4A-14, Z4A-17 and Z4A-20 had an extra cysteine residue in the unique domain II, while Z4A-15 lacked the second conserved cysteine residue in the unique domain I. The two fusion proteins of Z4A-15 and Z4A-20 were successfully detected by SDS-PAGE and Western blotting, although the protein level was relatively low. Based on the occurrence of the four major epitopes, as well as the lengths of the two glutamine repeats, 8, 6, and 8 genes were assigned to the Gli-2 loci on the respective chromosomes 6A, 6B, and 6D and a total of respectively 16, 0 and 23 immunogenic peptides were identified. In addition, 4 of the 5 genes with odd numbers of cysteine residues were assigned to chromosome 6D, suggesting that common wheat cultivar Zhengmai 004 has the potential to induce celiac disease(CD) and that the D genome exerts the most influence on gluten quality, but is the most deleterious for CD patients. By phylogenetic analysis, 11 exceptional α-gliadins with few or no immunogenic peptides from Triticum monococcum and Aegilops tauschii were detected, a finding that further supports the prospect of CD prevention. Finally, secondary structure prediction showed that most(98.48%) of the α-gliadins invariably contained five conserved α-helices(H1 to H5) in the two glutamine repeats and unique domains and 67.68% of the α-gliadins also contained a β-strand(S) in the C-terminal unique domains. An absent α-helix H2, 1–2 extra α-helices, or an additional β-strand(SE) also probably occurred in some cases. Of the 22 cloned genes in this work, 10 putative proteins contained 1–2 extra α-helices in addition to the five conserved α-helices or the additional β-strand. The observation that most of the α-helices and β-strands were present in the two unique domains and that an extra α-helix also probably occurred in the two glutamine repeats in some desirable genes strongly suggested that these two uniquedomains are the most important regions for the function of α-gliadins, although the glutamine repeats would also contribute in some cases.
基金Acknowledgments This work was supported by the National Natural Science Foundation of China (No. 31301037 and 31470012 to G.S.), the Natural Science Foundation of Yunnan Province of China (No. 2013FB068 to G.S.), the Young Academic and Technical Leader Raising Foundation of Yunnan Province (No. 2014HB046, to G.S.), the Western Light Talent Culture Project of the Chinese Academy of Sciences (to G.S.), and the Yunnan Recruitment Program of Experts in Sciences (2012HA016 to J.W.).
文摘Horizontal gene transfer(HGT) refers to the flow of genetic materials to non-offspring,and occasionally HGT in plants can improve the adaptation of organisms in new niches due to expanded metabolic capability.Anthocyanins are an important group of water-soluble red,purple,or blue secondary metabolites,whose diversity results from modification after the main skeleton biosynthesis.Cuscuta is a stem holoparasitic genus,whose members form direct connection with hosts to withdraw water,nutrients,and macromolecules.Such intimate association is thought to increase the frequency of HGT.By transcriptome screening for foreign genes in Cuscuta australis,we discovered that one gene encoding a putative anthocyanin acyltransferase gene of the BAHD family,which is likely to be involved in anthocyanin modification,was acquired by C.australis from Fabaceae through HGT.The anthocyanin acyltransferase-like(AT-like) gene was confirmed to be present in the genome assembly of C.australis and the transcriptomes of Cuscuta pentagona.The higher transcriptional level in old stems is consistent with its putative function in secondary metabolism by stabilizing anthocyanin at neutral pH and thus HGT of this AT-like gene may have improved biotic and abiotic resistance of Cuscuta.