ζ (zeta)-carotene desaturase (ZDS) is a key enzyme for carotenoid biosynthesis, demonstrating high association with the yellow pigment (YP) content in wheat grain. Cloning ZDS gene and developing functional mar...ζ (zeta)-carotene desaturase (ZDS) is a key enzyme for carotenoid biosynthesis, demonstrating high association with the yellow pigment (YP) content in wheat grain. Cloning ZDS gene and developing functional markers are important for marker-assisted selection in wheat breeding. In the present study, the full-length DNA sequence of a ZDS gene on wheat chromosome 2A, designated TaZds-A1, was cloned, with 14 exons and 13 introns, and it has an open reading frame (ORF) of 1 707 bp, encoding 568 amino acid residues. A co-dominant functional marker, YP2A-1, was designed based on the polymorphisms of two alleles at the locus, TaZds-Ala and TaZds-Alb, yielding 183- and 179-bp fragments in TaZds-Ala and TaZds-Alb genotypes, respectively. A new QTL for YP content was detected on chromosome 2A, co-segregating with the functional marker YP2A-1 and TaZds-A1; it explained 11.3% of the phenotypic variance for YP content in a doubled haploid (DH) population from Zhongyou 9507/CA9632. Among 217 Chinese wheat cultivars and advanced lines, the average grain YP content of 126 cultivars with TaZds-Alb allele was 7.8% higher than that of 91 cultivars with TaZds- Ala allele.展开更多
Considering the advantages of single nucleotide polymorphisms(SNP) in genotyping and variety identification, the first set public SNP markers at Cotton Marker Database(http://www.cottonmarker.org/) were validated...Considering the advantages of single nucleotide polymorphisms(SNP) in genotyping and variety identification, the first set public SNP markers at Cotton Marker Database(http://www.cottonmarker.org/) were validated and screened across standard varieties of cotton distinctness, uniformity and stability(DUS) test, aiming to obtain an appropriate set of core SNP markers suitable for upland cotton cultivars in China. A total of 399 out of 1 005 SNPs from 270 loci including 170 insertions-deletions(In Dels) were evaluated for their polymorphisms among 30 standard varieties using Sanger sequencing. As a result, 147 loci were sequenced successfully, 377 SNPs and 49 In Dels markers were obtained. Among the 377 SNP markers, 333 markers(88.3%) were polymorphic between Gossypium hirsutum and G. barbadense, while 164 markers(43.5%) were polymorphic within upland cotton. As for In Del markers, the polymorphic rate is relatively lower than that of SNP both between species and within species. The homozygous DNA locus ratio of 121 SNPs was higher than 86.2% while that of other 43 SNPs was less than 70%. Only 64 SNPs displayed completely homozygous genotypes among all of the detected upland cotton varieties with 100% homozygous DNA locus ratio. At last, a set of 23 pairs of core SNPs were achieved in view of avoidance of linkage, with polymorphism information content(PIC) values varying from 0.21 to 0.38 with an average of 0.28. Genotype characteristics and genetic diversity were analyzed based on the set of core markers, while 40 pairs of core simple-sequence repeats(SSR) primers comprised of 10 sets of four multiplex PCR combinations were also used for analysis based on fluorescence detection system. Comparison results indicated that the genetic diversity level was almost equal, while various varieties were significantly different from each other. Genetic relationship revealed by SSR markers is related to geographic source to a certain extent. Meanwhile clustering results analyzed by SNP markers are more consistent with kinship, which demonstrated that the screen strategy for core SNP marker is effective.展开更多
基金supported by the National Natural Science Foundation of China (30871522 and 30830072)the National Basic Research Program of China(2009CB118300)an Earmarked Fund for the Modern Agro-Industry Technology Research System, China (CARS-3-1-3)
文摘ζ (zeta)-carotene desaturase (ZDS) is a key enzyme for carotenoid biosynthesis, demonstrating high association with the yellow pigment (YP) content in wheat grain. Cloning ZDS gene and developing functional markers are important for marker-assisted selection in wheat breeding. In the present study, the full-length DNA sequence of a ZDS gene on wheat chromosome 2A, designated TaZds-A1, was cloned, with 14 exons and 13 introns, and it has an open reading frame (ORF) of 1 707 bp, encoding 568 amino acid residues. A co-dominant functional marker, YP2A-1, was designed based on the polymorphisms of two alleles at the locus, TaZds-Ala and TaZds-Alb, yielding 183- and 179-bp fragments in TaZds-Ala and TaZds-Alb genotypes, respectively. A new QTL for YP content was detected on chromosome 2A, co-segregating with the functional marker YP2A-1 and TaZds-A1; it explained 11.3% of the phenotypic variance for YP content in a doubled haploid (DH) population from Zhongyou 9507/CA9632. Among 217 Chinese wheat cultivars and advanced lines, the average grain YP content of 126 cultivars with TaZds-Alb allele was 7.8% higher than that of 91 cultivars with TaZds- Ala allele.
基金supported by the the Basic R&D Operation Special Fund for the Central Level, Non-Profit, Scientific Research Institutes, Ministry of Agriculture, China (1610162015A08)the National Key Technologies R&D Program of China during the 12th Five-Year Plan period (2011BAD35B09)
文摘Considering the advantages of single nucleotide polymorphisms(SNP) in genotyping and variety identification, the first set public SNP markers at Cotton Marker Database(http://www.cottonmarker.org/) were validated and screened across standard varieties of cotton distinctness, uniformity and stability(DUS) test, aiming to obtain an appropriate set of core SNP markers suitable for upland cotton cultivars in China. A total of 399 out of 1 005 SNPs from 270 loci including 170 insertions-deletions(In Dels) were evaluated for their polymorphisms among 30 standard varieties using Sanger sequencing. As a result, 147 loci were sequenced successfully, 377 SNPs and 49 In Dels markers were obtained. Among the 377 SNP markers, 333 markers(88.3%) were polymorphic between Gossypium hirsutum and G. barbadense, while 164 markers(43.5%) were polymorphic within upland cotton. As for In Del markers, the polymorphic rate is relatively lower than that of SNP both between species and within species. The homozygous DNA locus ratio of 121 SNPs was higher than 86.2% while that of other 43 SNPs was less than 70%. Only 64 SNPs displayed completely homozygous genotypes among all of the detected upland cotton varieties with 100% homozygous DNA locus ratio. At last, a set of 23 pairs of core SNPs were achieved in view of avoidance of linkage, with polymorphism information content(PIC) values varying from 0.21 to 0.38 with an average of 0.28. Genotype characteristics and genetic diversity were analyzed based on the set of core markers, while 40 pairs of core simple-sequence repeats(SSR) primers comprised of 10 sets of four multiplex PCR combinations were also used for analysis based on fluorescence detection system. Comparison results indicated that the genetic diversity level was almost equal, while various varieties were significantly different from each other. Genetic relationship revealed by SSR markers is related to geographic source to a certain extent. Meanwhile clustering results analyzed by SNP markers are more consistent with kinship, which demonstrated that the screen strategy for core SNP marker is effective.