Biochemical data bearing on the relationship between the genome of Triticum urartu and the A and B genomes of the polyploid wheats

To determine whether the Triticum urartu genome is more closely related to the A or B genome of the polyploid wheats, the amino acid sequence of its purothionin was compared to the amino acid sequences of the purothionins in Triticum monococcum, Triticum turgidum, and Triticum aestivum. The residue sequence of the purothionin from T. urartu differs by five and six amino acid substitutions respectively from the alpha 1 and alpha 2 forms coded for by genes in the B and D genomes, and is identical to the beta form specified by a gene in the A genome. Therefore, the T. urartu purothionin is either coded by a gene in the A genome or a chromosome set highly homologous to it. The results demonstrate that at least a portion of the T. urartu and T. monococcum genomes is homologous and probably identical. A variety of other studies have also shown that T. urartu is very closely related to T. monococcum and, in all likelihood, also possesses the A genome. Therefore, it could be argued that either T. urartu and T. monococcum are the same species or that T. urartu rather than T. monococcum is the source of the A genome in T. turgidum and T. aestivum. Except for Johnson's results, our data and that of others suggest a revised origin of polyploid wheats. Specifically, the list of six putative B genome donor species is reduced to five, all members of the Sitopsis section of the genus Aegilops.     

Molecular characterization of a diagnostic DNA marker for domesticated tetraploid wheat provides evidence for gene flow from wild tetraploid wheat to hexaploid wheat

All forms of domesticated tetraploid wheat (Triticum turgidum, genomes AABB) are nearly monomorphic for restriction fragment length polymorphism (RFLP) haplotype a at the Xpsr920 locus on chromosome 4A (Xpsr920-A1a), and wild tetraploid wheat is monomorphic for haplotype b. The Xpsr920-A1a/b dimorphism provides a molecular marker for domesticated and wild tetraploid wheat, respectively. Hexaploid wheat (Triticum aestivum, genomes AABBDD) is polymorphic for the 2 haplotypes. Bacterial artificial chromosome (BAC) clones hybridizing with PSR920 were isolated from Triticum urartu (genomes AA), Triticum monococcum (genomes AmAm), and T. turgidum ssp. durum (genomes AABB) and sequenced. PSR920 is a fragment of a putative ATP binding cassette (ABC) transporter gene (designated ABCT-1). The wheat ABCT-1 gene is more similar to the T. urartu gene than to the T. monococcum gene and diverged from the T. urartu gene about 0.7 MYA. The comparison of the sequence of the wheat A genome BAC clone with that of the T. urartu BAC clone provides the first insight into the microsynteny of the wheat A genome with that of T. urartu. Within 103 kb of orthologous intergenic space, 37 kb of new DNA has been inserted and 36 kb deleted leaving 49.7% of the region syntenic between the clones. The nucleotide substitution rate in the syntenic intergenic space has been 1.6 x 10(-8) nt(-1) year(-1), which is, respectively, 4 and 3 times as great as nucleotide substitution rates in the introns and the third codon positions of the juxtaposed gene. The RFLP is caused by a miniature inverted transposable element (MITE) insertion into intron 18 of the ABCT-A1 gene. Polymerase chain reaction primers were developed for the amplification of the MITE insertion site and its sequencing. The T. aestivum ABCT-A1a haplotype is identical to the haplotype of domesticated tetraploid wheat, and the ABCT-A1b haplotype is identical to that of wild tetraploid wheat. This finding shows for the first time that wild tetraploid wheat participated in the evolution of hexaploid wheat. A cline of the 2 haplotype frequencies exists across Euro-Asia in T. aestivum. It is suggested that T. aestivum in eastern Asia conserved the gene pool of the original T. aestivum more than wheat elsewhere.     

Microheterogeneity of the primary structure of the short repeated 5S DNA unit in diploid wheat Triticum monococcum L

Nucleotide sequences of two 5S rRNA genes located in repeated 327 bp long units were determined in diploid wheat Triticum monococcum. They were compared with sequences of 5S rRNA genes of Tr. monococcum and Tr. aestivum which were earlier determined. The differences were revealed in two localizations of the nucleotide sequence in 5S DNA coding regions of Tr. monococcum and - in nine localizations in nontranscribed spacer. It was established that the nucleotide sequence of 5S rRNA gene cloned in pTm5S9 plasmid and 5S DNA coding region in Tr. aestivum have significant homology. Diploid wheat Tr. monococcum was supposed to have 5S rRNA genes with different functional activity within one multigene family.     

Ribosomal RNA Multigene Loci: Nomads of the Triticeae Genomes

The nucleolus organizing regions (NORs) on the short arms of chromosomes 1A(m) and 5A(m) of diploid wheat, Triticum monococcum L., are at the most distal loci in the linkage maps of these two chromosome arms. This distal location differs from the interstitial location of the Nor loci on chromosome arms 1BS of tetraploid Triticum turgidum L. and hexaploid T. aestivum L., 5DS of T. aestivum and diploid Ae. tauschii Coss., and 5HS of barley. Moreover, the barley 5HS locus is at a different location than the 5DS locus. However, other markers, including the centromeres, are colinear. These findings showed that the major Nor loci have repeatedly changed position in the chromosome arms during the radiation of species in the tribe Triticeae without rearrangements of the linkage groups. It is suggested that Nor loci may change position via dispersion of minor loci, that are shown here to exist in the T. monococcum genome, magnification of gene copy numbers in these minor loci, and subsequent deletion of the original major loci. Implications of these findings for the use of rRNA nucleotide sequences in phylogenetic reconstructions are pointed out.     

Sequence Variations and Haplotype Identification of Wheat Dimeric α-Amylase Inhibitor Genes in Einkorn Wheats

This study characterizes 80 dimeric alpha-amylase inhibitor genes from 68 accessions of the einkorn wheats Triticum urartu, T. boeoticum, and T. monococcum. The mature protein coding sequences of WDAI genes were analyzed. Nucleotide sequence variations in these regions resulted from base substitution and/or indel mutations. Most of the WDAI gene sequences from T. boeoticum and all sequences from T. monococcum had one nucleotide insertion in the coding region, such that these alpha-amylase inhibitor sequences could not encode the correct mature proteins. We identified 21 distinct haplotypes from the diploid wheat WDAI gene sequences. A main haplotype was found in 15 gene samples from the A(u) genome and 35 gene samples from the A(m) genome. The T. monococcum and T. boeoticum accessions shared the same main haplotype, with 25 samples from T. monococcum and 10 from T. boeoticum. The WDAI gene sequences from the A(u) and A(m) genomes could be obviously clustered into two clades, but the sequences from the A(m) genome of T. boeoticum and T. monococcum could not be clearly distinguished. The phylogenetic analysis revealed that the WDAI gene sequences from the A(m) genome had accumulated fewer variations and evolved at a slower rate than the sequences from the A(u) genome. Although some accessions from only one or two areas had unique mutations at the same position, the diversity of WDAI gene sequences in diploid wheat showed little relationship to the origin of the accessions.     

Identification of variation in adaptively important traits and genome-wide analysis of trait-marker associations in Triticum monococcum

Einkorn wheat Triticum monococcum (2n=2x=14, A(m)A(m)) is one of the earliest domesticated crops. However, it was abandoned for cultivation before the Bronze Age and has infrequently been used in wheat breeding. Little is known about the genetic variation in adaptively important biological traits in T. monococcum. A collection of 30 accessions of diverse geographic origins were characterized for phenotypic variation in various agro-morphological traits including grain storage proteins and endosperm texture, nucleotide-binding site (NBS) domain profiles of resistance (R) genes and resistance gene analogues (RGAs), and germination under salt and drought stresses. Forty-six SSR (single sequence repeat) markers from bread wheat (T. aestivum, 2n=6x=42, AABBDD) A genome were used to establish trait-marker associations using linear mixed models. Multiple significant associations were identified, some of which were on chromosomal regions containing previously known genetic loci. It is concluded that T. monococcum possesses large genetic diversity in multiple traits. The findings also indicate that the efficiency of association mapping is much higher in T. monococcum than in other plant species. The use of T. monococcum as a reference species for wheat functional genomics is discussed.     

The specific isolation of complete 5S rDNA units from chromosome 1A of hexaploid, tetraploid, and diploid wheat species using PCR with head-to-head oriented primers.

The presence of 5S rDNA units on chromosome 1A of Triticum aestivum was shown by the development of a specific PCR test, using head-to-head oriented primers. This primer set allowed the amplification of complete 5S DNA units and was used to isolate SS-Rrna-A1 sequences from polyploid and diploid wheat species. Multiple-alignment and parsimony analyses of the 132 sequences divided the sequences into four types. The isolates from T. aestivum and the tetraploid species (T. dicoccoides, T. dicoccum, T durum, T. araraticum, and T timopheevi) were all of one type, which was shown to be closely related to the type mainly characteristic for T. urartu. The other two types were isolated exclusively from the diploid species T. monococcum, T aegilopoides, T. thaoudar, and T. sinskajae and the hexaploid species T. zhukovski. Triticum monococcum was the only species for which representatives of each of the four sequence types were found to be present. Further, we discuss the possible multicluster arrangement of the 5S-Rrna-A1 array.     

Isolation of a cDNA encoding a granule-bound 152-kilodalton starch-branching enzyme in wheat

Screening of a wheat (Triticum aestivum) cDNA library for starch-branching enzyme I (SBEI) genes combined with 5'-rapid amplification of cDNA ends resulted in isolation of a 4,563-bp composite cDNA, Sbe1c. Based on sequence alignment to characterized SBEI cDNA clones isolated from plants, the SBEIc predicted from the cDNA sequence was produced with a transit peptide directing the polypeptide into plastids. Furthermore, the predicted mature form of SBEIc was much larger (152 kD) than previously characterized plant SBEI (80-100 kD) and contained a partial duplication of SBEI sequences. The first SBEI domain showed high amino acid similarity to a 74-kD wheat SBEI-like protein that is inactive as a branching enzyme when expressed in Escherichia coli. The second SBEI domain on SBEIc was identical in sequence to a functional 87-kD SBEI produced in the wheat endosperm. Immunoblot analysis of proteins produced in developing wheat kernels demonstrated that the 152-kD SBEIc was, in contrast to the 87- to 88-kD SBEI, preferentially associated with the starch granules. Proteins similar in size and recognized by wheat SBEI antibodies were also present in Triticum monococcum, Triticum tauschii, and Triticum turgidum subsp. durum.     

Genome analysis at different ploidy levels allows cloning of the powdery mildew resistance gene Pm3b from hexaploid wheat

In wheat, race-specific resistance to the fungal pathogen powdery mildew (Blumeria graminis f. sp. tritici) is controlled by the Pm genes. There are 10 alleles conferring resistance at the Pm3 locus (Pm3a to Pm3j) on chromosome 1AS of hexaploid bread wheat (Triticum aestivum L.). The genome of hexaploid wheat has a size of 1.6 x 1010 bp and contains more than 80% of repetitive sequences, making positional cloning difficult. Here, we demonstrate that the combined analysis of genomes from wheat species with different ploidy levels can be exploited for positional cloning in bread wheat. We have mapped the Pm3b gene in hexaploid wheat to a genetic interval of 0.97 centimorgan (cM). The diploid T. monococcum and the tetraploid T. turgidum ssp. durum provided models for the A genome of hexaploid wheat and allowed to establish a physical contig spanning the Pm3 locus. Although the haplotypes at the Pm3 locus differed markedly between the three species, a large resistance gene-like family specific to wheat group 1 chromosomes was consistently found at the Pm3 locus. A candidate gene for Pm3b was identified using partial sequence conservation between resistant line Chul and T. monococcum cv. DV92. A susceptible Pm3b mutant, carrying a single-base pair deletion in the coding region of the candidate gene was isolated. When tested in a single cell transformation assay, the Pm3b candidate gene conferred race-specific resistance to powdery mildew. These results demonstrate that the candidate gene, a member of the coiled-coil nucleotide binding site leucine-rich repeat (NBS-LRR) type of disease resistance genes, is the Pm3b gene.     

小麦BBC1基因cDNA的克隆与分析

从小麦(Triticum aestivum L．)中克隆了一个BBC1基因的cDNA。分析结果表明,该基因编码一亲水多肽,富含丙氨酸、赖氨酸、精氮酸和谷氦酸。该基因的转录受低温调控。在小麦基因组中,BBC1基因以一个小家族的形式存在。     

小麦BBC1基因cDNA的克隆与分析

从小麦(Triticum aestivum L.)中克隆了一个BBC1基因的cDNA.分析结果表明,该基因编码一亲水多肽,富含丙氨酸、赖氨酸、精氨酸和谷氨酸.该基因的转录受低温调控.在小麦基因组中,BBC1基因以一个小家族的形式存在.     

The evolution of polyploid wheats: identification of the A genome donor species.

Cytogenetic work has shown that the tetraploid wheats, Triticum turgidum and T. timopheevii, and the hexaploid wheat T. aestivum have one pair of A genomes, whereas hexaploid T. zhukovskyi has two. Variation in 16 repeated nucleotide sequences was used to identify sources of the A genomes. The A genomes of T. turgidum, T. timopheevii, and T. aestivum were shown to be contributed by T. urartu. Little divergence in the repeated nucleotide sequences was detected in the A genomes of these species from the genome of T. urartu. In T. zhukovskyi one A genome was contributed by T. urartu and the other was contributed by T. monococcum. It is concluded that T. zhukovskyi originated from hybridization of T. timopheevii with T. monococcum. The repeated nucleotide sequence profiles in the A genomes of T. zhukovskyi showed reduced correspondence with those in the genomes of both ancestral species, T. urartu and T. monococcum. This differentiation is attributed to heterogenetic chromosome pairing and segregation among chromosomes of the two A genomes in T. zhukovskyi.     

小麦A，B和D基因组同源DNA序列在进化过程中的演变研究

选取已定位的大麦1H染色体的STS标记NWG913为引物,在普通小麦（Tritium aestivum L.）及其4个可能的起源种乌拉尔图小麦（T.urartu T.)、栽培一粒小麦（T.monococcum.L)栽培二粒小麦（T.dicoccum S.）、方穗山羊草（Ae.squarrosa L.)上特异性扩增。扩增产物克隆测序后对其进行序列分析,由序列差异的程度来确定这几个物种之间的亲缘关系。实验结果表明,普通小麦（Tritium aestivum L.)的A基因组此段序列与乌拉尔图小麦（T.urartu T.)、栽培一粒小麦（T.monococcum L.)、栽培二粒小麦（T.dicoccum S.)A基因组此段序列完全相同;普通小麦的D基因组此段序列与方穗山羊草（Ae.squarrosa L.)也完全相同;普通小麦的B基因组此段序列和栽培二粒小麦B基因组此段序列有0．61％的差异。研究结果一方面对现有的普通小麦A、B、D基因组起源和进化理论给予了分子水平上的证明,同时也揭示了同一物种不同的基因组化进化速度存在差异。     

Inheritance of dense spike in diploid wheat and Aegilops squarrosa

The individuals of diploid wheat Triticum boeoticum, T. monococcum and T. sinskajae and goatgrass Aegilops squarrosa were picked out with screening the dense spike characteristics. The dense-spike accessions were discovered in diploid wheat (T. sinskajae) and Ae. squarrosa. Inheritance of the dense spike was studied. The trait was found to be controlled by a recessive gene in T. sinskajae and by an incomplete dominant gene in Ae. squarrosa. The dosage effect of dominant gene C was detected in interspecific pentaploid F1 hybrid plants T. compactum x T. palmovae (2n =35, A(u)A(b)BDD genome). The spike of pentaploid hybrid was not so dense as compared to hexaploid wheat T. compactum. This is the first report showing similarity of the expression of dominant gene C on D genome of the hexaploid wheat to that of dense spike gene in Ae. squarrosa. The existence of dense-spike accessions of Ae. squarrosa allows us to hypothesize that the origin of T. compactum is independent from that of common wheat.     

Genetic Map of Diploid Wheat, Triticum Monococcum L., and Its Comparison with Maps of Hordeum Vulgare L

A genetic map of diploid wheat, Triticum monococcum L., involving 335 markers, including RFLP DNA markers, isozymes, seed storage proteins, rRNA, and morphological loci, is reported. T. monococcum and barley linkage groups are remarkably conserved. They differ by a reciprocal translocation involving the long arms of chromosomes 4 and 5, and paracentric inversions in the long arm of chromosomes 1 and 4; the latter is in a segment of chromosome arm 4L translocated to 5L in T. monococcum. The order of the markers in the inverted segments in the T. monococcum genome is the same as in the B and D genomes of T. aestivum L. The T. monococcum map differs from the barley maps in the distribution of recombination within chromosomes. The major 5S rRNA loci were mapped on the short arms of T. monococcum chromosomes 1 and 5 and the long arms of barley chromosomes 2 and 3. Since these chromosome arms are colinear, the major 5S rRNA loci must be subjected to positional changes in the evolving Triticeae genome that do not perturb chromosome colinearity. The positional changes of the major 5S rRNA loci in Triticeae genomes are analogous to those of the 18S-5.8S-26S rRNA loci.     

Rapid genome divergence at orthologous low molecular weight glutenin loci of the A and Am genomes of wheat

To study genome evolution in wheat, we have sequenced and compared two large physical contigs of 285 and 142 kb covering orthologous low molecular weight (LMW) glutenin loci on chromosome 1AS of a diploid wheat species (Triticum monococcum subsp monococcum) and a tetraploid wheat species (Triticum turgidum subsp durum). Sequence conservation between the two species was restricted to small regions containing the orthologous LMW glutenin genes, whereas >90% of the compared sequences were not conserved. Dramatic sequence rearrangements occurred in the regions rich in repetitive elements. Dating of long terminal repeat retrotransposon insertions revealed different insertion events occurring during the last 5.5 million years in both species. These insertions are partially responsible for the lack of homology between the intergenic regions. In addition, the gene space was conserved only partially, because different predicted genes were identified on both contigs. Duplications and deletions of large fragments that might be attributable to illegitimate recombination also have contributed to the differentiation of this region in both species. The striking differences in the intergenic landscape between the A and A(m) genomes that diverged 1 to 3 million years ago provide evidence for a dynamic and rapid genome evolution in wheat species.     

灌浆期水分亏缺条件下二、四、六倍体小麦收获指数和水分利用效率的演化

为揭示不同倍性小麦生长发育、产量性状及水分利用对灌浆期水分亏缺响应的差异,选用二倍体野生一粒小麦(Triticum boeoticum)、栽培一粒小麦(T.monococcum),四倍体野生二粒小麦(T.dicoccoides)、栽培二粒小麦(T.dicoccon),和两个普通六倍体小麦(T.aestivum)品种‘长武134’和‘陕253’6个小麦品种作为供试材料。采用盆栽控水的方法,测定和分析了不同灌浆期土壤水分条件下小麦株高、旗叶叶面积、穗长、根干重、地上生物量、根冠比、千粒重、粒数、产量、收获指数、蒸腾耗水量和水分利用效率等性状的变化。在小麦染色体倍体由二倍体向六倍体进化的过程中,小麦地上生物量、千粒重、穗粒数、产量、收获指数和水分利用效率都显著增加。随着土壤水分从正常→中度亏缺→重度亏缺的减少,收获指数先增大后减小,分别为41.26%、42.48%和38.19%;生物量水分利用效率逐渐增大,分别为2.39、2.43和2.53g·kg–1;产量水分利用效率分别为1.05、1.10和1.04g·kg–1。在灌浆期水分条件是影响收获指数和水分利用效率的关键因素之一。灌浆期的水分亏缺有利于六倍体小麦的收获指数和四倍体的生物量水分利用效率的提高。中度的水分亏缺有利于四倍体和六倍体产量水分利用效率的提高。