Shou-Fen Dai

Characterization of two HMW glutenin subunit genes from Taenitherum Nevski

The compositions of high molecular weight (HMW) glutenin subunits from three species of Taenitherum Nevski (TaTa, 2n = 2x = 14), Ta. caput-medusae, Ta. crinitum and Ta. asperum, were investigated by SDS-PAGE analysis. The electrophoresis mobility of the x-type HMW glutenin subunits were slower or equal to that of wheat HMW glutenin subunit Dx2, and the electrophoresis mobility of the y-type subunits were faster than that of wheat HMW glutenin subunit Dy12. Two HMW glutenin genes, designated as Tax and Tay, were isolated from Ta. crinitum, and their complete nucleotide coding sequences were determined. Sequencing and multiple sequences alignment suggested that the HMW glutenin subunits derived from Ta. crinitum had the similar structures to the HMW glutenin subunits from wheat and related species with a signal peptide, and N- and C-conservative domains flanking by a repetitive domain consisted of the repeated short peptide motifs. However, the encoding sequences of Tax and Tay had some novel modification compared with the HMW glutenin genes reported so far: (1) A short peptide with the consensus sequences of KGGSFYP, which was observed in the N-terminal of all known HMW glutenin genes, was absent in Tax; (2) There is a specified short peptide tandem of tripeptide, hexapeptide and nonapeptide and three tandem of tripeptide in the repetitive domain of Tax; (3) The amino acid residues number is 105 (an extra Q presented) but not 104 in the N-terminal of Tay, which was similar to most of y-type HMW glutenin genes from Elytrigia elongata and Crithopsis delileana. Phylogenetic analysis indicated that Tax subunit was mostly related to Ax1, Cx, Ux and Dx5, and Tay was more related to Ay, Cy and Ry.

Quantitative Trait Loci Associated with Micronutrient Concentrations in Two Recombinant Inbred Wheat Lines

Micronutrient malnutrition affects over three billion people worldwide, especially women and children in developing countries. Increasing the bioavailable concentrations of essential elements in the edible portions of crops is an effective resolution to address this issue. To determine the genetic factors controlling micronutrient concentration in wheat, the quantitative trait locus (QTL) analysis for iron, zinc, copper, manganese, and selenium concentrations in two recombinant inbred line populations was performed. In all, 39 QTLs for five micronutrient concentrations were identified in this study. Of these, 22 alleles from synthetic wheat SHW-L1 and seven alleles from the progeny line of the synthetic wheat Chuanmai 42 showed an increase in micronutrient concentrations. Five QTLs on chromosomes 2A, 3D, 4D, and 5B found in both the populations showed significant phenotypic variation for 2–3 micronutrient concentrations. Our results might help understand the genetic control of micronutrient concentration and allow the utilization of genetic resources of synthetic hexaploid wheat for improving micronutrient efficiency of cultivated wheat by using molecular marker-assisted selection.

Formation of unreduced gametes is impeded by homologous chromosome pairing in tetraploid Triticum turgidum × Aegilops tauschii hybrids

It is believed that unreduced gametes with somatic chromosome numbers play a predominant role in natural polyploidization. Allohexaploid bread wheat originated from spontaneous hybridization of Triticum turgidum L. with Aegilops tauschii Coss. Unreduced gametes originating via meiotic restitution, including first-division restitution (FDR) and single-division meiosis (SDM), are well documented in triploid F1 hybrids of T. turgidum with diploid Ae. tauschii (genomic constitution ABD, usually with 21 univalents in meiotic metaphase I). In this study, two T. turgidum lines known to carry genes for meiotic restitution were crossed to tetraploid Ae. tauschii. The resulting F1 hybrids (genomes ABDD), had seven pairs of homologous chromosomes and regularly formed 14 univalents and seven bivalents at metaphase I. Neither FDR nor SDM were observed. The distribution of chromosome numbers among progeny obtained by self pollination and a backcross to T. turgidum showed the absence of unreduced gametes. These results suggest that high homologous pairing interfered with meiotic restitution and the formation of unreduced gametes. This may be related to asynchronous movement during meiosis between paired and unpaired chromosomes or to uneven distribution of chromosomes in anaphases, resulting in nonviable gametes.

The Crossability of Triticum turgidum with Aegilops tauschii

Two experiments to investigate the crossability of Triticum turgidum with Aegilops tauschii are described. In the first experiment, 372 wide hybridization combinations were done by crossing 196 T. turgidum lines belonging to seven subspecies with 13 Ae. tauschii accessions. Without embryo rescue and hormone treatment, from the 66220 florets pollinated, 3713 seeds were obtained, with a mean crossability percentages of 5.61% which ranged from 0 to 75%. A lot of hybrid seeds could germinate and produce plants. Out of 372 combinations, 73.12% showed a very low crossability less than 5%, 23.39% showed the crossability of 5–30%, 2.69% showed the crossability of 30–50%, 0.81% showed high crossability more than 50%, respectively. Among the seven T. turgidum subspecies, there were significant differences in crossability. The ssp. dicoccoides and dicoccon showed the highest crossability, while polonicum the lowest. All the crossability percentages more than 30% were obtained from the crossing of ssp. dicoccoides or...

Phylogenetic inferences in Avena based on analysis of FL intron2 sequences

The development and application of molecular methods in oats has been relatively slow compared with other crops. Results from the previous analyses have left many questions concerning species evolutionary relationships unanswered, especially regarding the origins of the B and D genomes, which are only known to be present in polyploid oat species. To investigate the species and genome relationships in genus Avena, among 13 diploid (A and C genomes), we used the second intron of the nuclear gene FLORICAULA/LEAFY (FL int2) in seven tetraploid (AB and AC genomes), and five hexaploid (ACD genome) species. The Avena FL int2 is rather long, and high levels of variation in length and sequence composition were found. Evidence for more than one copy of the FL int2 sequence was obtained for both the A and C genome groups, and the degree of divergence of the A genome copies was greater than that observed within the C genome sequences. Phylogenetic analysis of the FL int2 sequences resulted in topologies that contained four major groups; these groups reemphasize the major genomic divergence between the A and C genomes, and the close relationship among the A, B, and D genomes. However, the D genome in hexaploids more likely originated from a C genome diploid rather than the generally believed A genome, and the C genome diploid A. clauda may have played an important role in the origination of both the C and D genome in polyploids.

Genome-Wide Quantitative Trait Locus Mapping Identifies Multiple Major Loci for Brittle Rachis and Threshability in Tibetan Semi-Wild Wheat (Triticum aestivum ssp. tibetanum Shao)

Tibetan semi-wild wheat (Triticum aestivum ssp. tibetanum Shao) is a semi-wild hexaploid wheat resource that is only naturally distributed in the Qinghai-Tibet Plateau. Brittle rachis and hard threshing are two important characters of Tibetan semi-wild wheat. A whole-genome linkage map of T. aestivum ssp. tibetanum was constructed using a recombinant inbred line population (Q1028×ZM9023) with 186 lines, 564 diversity array technology markers, and 117 simple sequence repeat markers. Phenotypic data on brittle rachis and threshability, as two quantitative traits, were evaluated on the basis of the number of average spike rachis fragments per spike and percent threshability in 2012 and 2013, respectively. Quantitative trait locus (QTL) mapping performed using inclusive composite interval mapping analysis clearly identified four QTLs for brittle rachis and three QTLs for threshability. However, three loci on 2DS, 2DL, and 5AL showed pleiotropism for brittle rachis and threshability; they respectively explained 5.3%, 18.6%, and 18.6% of phenotypic variation for brittle rachis and 17.4%, 13.2%, and 35.2% of phenotypic variation for threshability. A locus on 3DS showed an independent effect on brittle rachis, which explained 38.7% of the phenotypic variation. The loci on 2DS and 3DS probably represented the effect of Tg and Br1, respectively. The locus on 5AL was in very close proximity to the Q gene, but was different from the predicted q in Tibetan semi-wild wheat. To our knowledge, the locus on 2DL has never been reported in common wheat but was prominent in T. aestivum ssp. tibetanum accession Q1028. It remarkably interacted with the locus on 5AL to affect brittle rachis. Several major loci for brittle rachis and threshability were identified in Tibetan semi-wild wheat, improving the understanding of these two characters and suggesting the occurrence of special evolution in Tibetan semi-wild wheat.

Evaluation on Chinese Bread Wheat Landraces for Low pH and Aluminum Tolerance Using Hydroponic Screening

Abstract Aluminum (Al) toxicity often takes place in acidic soils with a pH of 5.5 or lower. Breeding and cultivation of Al tolerance wheat can partially protect wheat escaping from Al toxicity. The scarcity of the tolerant sources impedes the wheat breeding. In order to find new Al tolerance sources, we screened 173 bread wheat landraces from Tibet of China using hydroponic screening. It was indicated that: (1) There were diversities on the root regenerate length (RRL). The RRL of a large of landraces were longer than 7.00 cm in pH 7 (58.38%) and pH 4.5 (66.47%), but shorter than 5.00 cm in pH 4.5 + 50 μM Al 3+ (80.93%). The low pH showed either promotion or restraining effects depend on landraces, but Al toxicity under low pH only showed restraining effects on the root elongation. (2) There were also diversities on root tolerance index of low pH (RTI1) or root aluminum tolerance index (RTI2) among cultivars. The RTI1 varied from a narrow range but with relatively high value (0.8722-1.2953) in comparison with that of RTI2 (0.3829-1.0058), and the RTI1 of approximately 60% landraces was higher than 1.0000, the RTI2 of only 19.07% landraces was higher than 0.7000, suggesting that Al toxicity acted as an important factor for the reduction of the root elongation under acidic soils. (3) The RTI1 of many wheats was higher than 1.0000, and As2256 and As2295 were the most tolerant for low pH, with RTI1 1.2953 and 1.2925, respectively. (4) Based on RTI2, seven wheats showed similar or higher tolerance to Al toxicity than Chinese Spring (CS), a known tolerance wheat. Much better tolerance existed in landraces of As1543 and As1242, which can be used as the new parents for Al tolerant breeding.

Characterization of novel HMW-GS in two diploid species of Eremopyrum.

Three HMW-GS and the respective ORFs from diploid species Eremopyrum distans and Eremopyrum triticeum were characterized. Compared to homologous proteins, they showed novel modifications in all domains. In the N-terminals, the y subunit from Er. triticeum (Xey) had 98 aa residues. A short G/IIFWGTS peptide deletion was responsible for the reduced number of aa residues. The end peptide in the y subunit from Er. distans (Fy) was IPTLLR. This unique structure was involved in a replacement between x types with IPA/TLLK/R and y types with R/TSSQTVQ. Both y subunits share the same short peptide LAAQLPAMCRL as x types in the C-terminals. Phylogenic relationships among orthologous genes from Triticeae species revealed that Fy and Xey were neither purely x type nor purely y type based on the N and C terminal residues. Divergence times indicated that Glu-Xe1 and Glu-F1 were separated from each other and that Glu-Xe1 separated from orthologous loci of wild wheat relatives earlier than Glu-F1. Based on the divergence times among Glu-F1, Glu-Xe1, Glu-O1, Glu-St1, and Glu-Ta1, it is possible that genome F separation from O, St, and Ta in species of Henrardia persica, Pseudoroegneria stipifolia, and Taeniatherum crinitum was more recent than the separation of F and Xe.

Characterization of HMW-GS genes Dx5 t and Dy12 t from Aegilops tauschii accession with subunit combination Dx5 t + Dy12 t

Aegilops tauschii is the generally accepted D genome diploid donor of hexaploid wheat. The significance of Ae. tauschii HMW-GS genes on bread-making properties of bread wheat has been well documented. Among them, Ae. tauschii HMW-GS Dx5 t +Dy12 t was thought as the pair with potentially value in endowing synthetic hexaploid wheat with good end-use qualities. In this paper, we isolated and sequenced genes Dx5 t and Dy12 t from Ae. tauschii accession As63. Amino acid sequence comparison indicated that Dy12 t from Ae. tauschii is more similar to Dy10 rather than Dy12 of bread wheat. The sequence of Dx5 t in Ae. tauschii accession As63 showed higher similarity to that of Dx5 in bread wheat than others. However, it is notable that Dx5 t lacked the additional cysteine residue in Dx5, which is responsible for good bread-making quality in common wheat. Moreover, compared to Dx5, Dx5 t has an extra hexpeptide repetitive motif unit (SGQGQQ) as well as five amino acid substitutions.

Analysis of high-molecular-weight glutenin subunits in five amphidiploids and their parental diploid species Aegilops umbellulata and Aegilops uniaristata

Amphidiploids serve as a bridge for transferring genes from wild species into wheat. In this study, five amphidiploids with AABBUU and AABBNN genomes were produced by spontaneous chromosome doubling of unreduced triploid F-1 gametes from crosses between diploid Aegilops (A. umbellulata accessions CIae 29 and PI 226500, and A. uniaristata accession PI 554419) and tetraploid Triticum turgidum (ssp. durum cultivar Langdon and ssp. dicoccum accessions PI 94668 and PI 349045) species. The composition of high-molecular-weight glutenin subunits (HMW-GS) in these amphidiploids and in their parental A. umbellulata and A. uniaristata species was analysed. As expected, the amphidiploids from T. turgidum ssp. dicoccum accession PI 944668 or PI 349045 and A. umbellulata accession CIae 29 or PI 226500 and A. uniaristata accession PI 554419 showed the same HMW-GS patterns as those of their Aegilops parents, because HMW-GS genes were all silenced in the T. turgidum ssp. dicoccum parents. The amphidiploids from CIae 29 and Langdon inherited all of the HMW-GS genes from their parents except for the Uy type. Using 10 and 15% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and 10% urea/SDS-PAGE, 11 Ux and ten Uy types in 16 combinations were observed in 48 A. umbellulata accessions, and two Nx and two Ny types in two combinations were detected in six A. uniaristata accessions. These novel HMW-GS variants may provide new genetic resources for improving the quality of wheat.