Dengcai Liu

mRNA and Small RNA Transcriptomes Reveal Insights into Dynamic Homoeolog Regulation of Allopolyploid Heterosis in Nascent Hexaploid Wheat

Newly synthesized allohexaploid wheat lines, which exhibit hybrid vigor or heterosis, may recapitulate the initial genetic state of common wheat. This work shows that, in addition to nonadditive expression, parental expression-level dominance of subgenomes in nascent allohexaploid wheat may contribute distinctively to heterosis through dynamic small RNA–mediated homoeolog regulations. Nascent allohexaploid wheat may represent the initial genetic state of common wheat (Triticum aestivum), which arose as a hybrid between Triticum turgidum (AABB) and Aegilops tauschii (DD) and by chromosome doubling and outcompeted its parents in growth vigor and adaptability. To better understand the molecular basis for this success, we performed mRNA and small RNA transcriptome analyses in nascent allohexaploid wheat and its following generations, their progenitors, and the natural allohexaploid cultivar Chinese Spring, with the assistance of recently published A and D genome sequences. We found that nonadditively expressed protein-coding genes were rare but relevant to growth vigor. Moreover, a high proportion of protein-coding genes exhibited parental expression level dominance, with genes for which the total homoeolog expression level in the progeny was similar to that in T. turgidum potentially participating in development and those with similar expression to that in Ae. tauschii involved in adaptation. In addition, a high proportion of microRNAs showed nonadditive expression upon polyploidization, potentially leading to differential expression of important target genes. Furthermore, increased small interfering RNA density was observed for transposable element–associated D homoeologs in the allohexaploid progeny, which may account for biased repression of D homoeologs. Together, our data provide insights into small RNA–mediated dynamic homoeolog regulation mechanisms that may contribute to heterosis in nascent hexaploid wheat.

Meiotic restriction in emmer wheat is controlled by one or more nuclear genes that continue to function in derived lines

Highly fertile F1 hybrids were made between Triticum turgidum L. ssp. turgidum (2nxa0=xa028, AABB) and Aegilops tauschii Coss. (2nxa0=xa014, DD) without embryo rescue and hormone treatment. The F1 plants had an average seed set of 25%. Approximately 96% of the F2 seeds were able to germinate normally and about 67% of the F2 plants were spontaneous amphidiploid (2nxa0=xa042, AABBDD). Cytological analysis of male gametogenesis of the F1 plants showed that meiotic restitution is responsible for the high fertility. A mitosis-like meiosis led to meiotic restitution at either of the two meiotic divisions resulting in unreduced gametes. Test crosses of the T. t. turgidum–Ae. tauschii amphidiploid with Ae. variabilis and rye suggested that the mitosis-like meiosis is controlled by one or more nuclear genes that continue to function in derived lines. This discovery indicates a potential application of such genes in producing double haploids.

Rapid changes of microsatellite flanking sequence in the allopolyploidization of new synthesized hexaploid wheat

It was suggested that the rapid changes of DNA sequence and gene expression occurred at the early stages of allopolyploid formation. In this study, we revealed the microsatellite (SSR) differences between newly formed allopolyploids and their donor parents by using 21 primer sets specific for D genome of wheat. It was indicated that rapid changes had occurred in the “shock” process of the allopolyploid formation between tetraploid wheat and Aegilops tauschii. The changes of SSR flanking sequence resulted in appearance of novel bands or disappearance of parental bands. The disappearance of the parental bands showed much higher frequencies in comparison with that of appearance of novel bands. Disappearance of the parental bands was not random. The frequency of disappearance in tetraploid wheat was much higher than in Ae. tauschii, i. e. the disappearance frequency in AABB genome was much higher than in D genome. Changes of SSR flanking sequence occurred at the early stage of F1 hybrid or just after chromosome doubling. From the above results, it can be inferred that SSR flanking sequence region was very active and was amenable to change in the process of polyploidization. This suggested that SSR flanking sequence probably had special biological function at the early stage of ployploidization. The rapid and directional changes at the early stage of polyploidization might contribute to the rapid evolution of the newly formed allopolyploid and allow the divergent genomes to act in harmony.

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.

Production of aneuhaploid and euhaploid sporocytes by meiotic restitution in fertile hybrids between durum wheat Langdon chromosome substitution lines and Aegilops tauschii

Fertile F(1) hybrids were obtained between durum wheat (Triticum durum Desf.) Langdon (LDN) and its 10 disomic substitution (LDN DS) lines with Aegilops tauschii accession AS60 without embryo rescue. Selfed seedset rates for hybrids of LDN with AS60 were 36.87% and 49.45% in 2005 and 2006, respectively. Similar or higher selfed seedset rates were observed in the hybrids of 1D (1A), 1D (1B), 3D (3A), 4D (4B), 7D (7A), and 2D (2B) with AS60, while lower in hybrids of 3D (3B) + 3BL, 5D (5A) + 5AL, 5D (5B) + 5B and 6D (6B) + 6BS with AS60 compared with the hybrids of LDN with AS60. Observation of male gametogenesis showed that meiotic restitution, both first-division restitution (FDR) and single-division meiosis (SDM) resulted in the formation of functional unreduced gametes, which in turn produced seeds. Both euhaploid and aneuhaploid gametes were produced in F(1) hybrids. This suggested a strategy to simultaneously transfer and locate major genes from the ancestral species T. turgidum or Ae. tauschii. Moreover, there was no significant difference in the aneuhaploid rates between the F(1) hybrids of LDN and LDN DS lines with AS60, suggesting that meiotic pairing between the two D chromosomes in the hybrids of LDN DS lines with AS60 did not promote the formation of aneuhaploid gametes.

Making the Bread: Insights from Newly Synthesized Allohexaploid Wheat

Bread wheat (or common wheat, Triticum aestivum) is an allohexaploid (AABBDD, 2n = 6x = 42) that arose by hybridization between a cultivated tetraploid wheat T.xa0turgidum (AABB, 2n = 4x = 28) and the wild goatgrass Aegilops tauschii (DD, 2n = 2x = 14). Polyploidization provided niches for rigorous genome modification at cytogenetic, genetic, and epigenetic levels, rendering a broader spread than its progenitors. This review summarizes the latest advances in understanding gene regulation mechanisms in newly synthesized allohexaploid wheat and possible correlation with polyploid growth vigor and adaptation. Cytogenetic studies reveal persistent association of whole-chromosome aneuploidy with nascent allopolyploids, in contrast to the genetic stability in common wheat. Transcriptome analysis of the euploid wheat shows that small RNAs are driving forces for homoeo-allele expression regulation via genetic and epigenetic mechanisms. The ensuing non-additively expressed genes and those with expression level dominance to the respective progenitor may play distinct functions in growth vigor and adaptation in nascent allohexaploid wheat. Further genetic diploidization of allohexaploid wheat is not random. Regional asymmetrical gene distribution, rather than subgenome dominance, is observed in both synthetic and natural allohexaploid wheats. The combinatorial effects of diverged genomes, subsequent selection of specific gene categories, and subgenome-specific traits are essential for the successful establishment of common wheat.

HMW-glutenin and gliadin variations in Tibetan weedrace, Xinjiang rice wheat and Yunnan hulled wheat

Nine Tibetan weedrace, 9 Xingjiang rice wheat and 14 Yunnan hulled wheat accessions were evaluated for the variability of HMW-glutenins and gliadins. Higher variability was observed for both HMW-glutenins and gliadins in Tibetan weedrace and Xingjiang rice wheat, while lower variability was observed in Yunan hulled wheat. There were 4 HMW-glutenin and 9 gliadin patterns in 9 Tibetan weedrace accessions, 5 HMW-glutenin and 8 gliadin patterns in 9 Xingjiang rice wheat accessions, and 3 HMW-glutenin and 8 gliadin patterns in 14 Yunnan hulled wheat accessions. In Xinjiang rice wheat, one accession (i.e. Daomai 2) carried subunits 2.1 + 10.1 encoded by Glu-D1, which is very rare in common wheat.

QTL Mapping for Important Agronomic Traits in Synthetic Hexaploid Wheat Derived from Aegiliops tauschii ssp. tauschii

Aegiliops tauschii is classified into two subspecies: Ae. tauschii ssp. tauschii and Ae. tauschii ssp. strangulata. Novel genetic variations exist in Ae. tauschii ssp. tauschii that can be utilized in wheat improvement. We synthesized a hexaploid wheat genotype (SHW-L1) by crossing an Ae. tauschii ssp. tauschii accession (AS60) with a tetraploid wheat genotype (AS2255). A population consisting of 171 F8 recombinant inbred lines was developed from SHW-L1 and Chuanmai 32 to identify QTLs associated with agronomic traits. A new genetic map with high density was constructed and used to detect the QTLs for heading date, kernel width, spike length, spikelet number, and thousand kernel weight. A total of 30 putative QTLs were identified for five investigated traits. Thirteen QTLs were located on D genomes of SHW-L1, six of them showed positive effect on agronomic traits. Chromosome region flanked by wPt-6133–wPt-8134 on 2D carried five environment-independent QTLs. Each QTL accounted for more than 10% phenotypic variance. These QTLs were highly consistent across environments and should be used in wheat breeding.

Mapping stripe rust resistance gene YrSph derived from Tritium sphaerococcum Perc. with SSR, SRAP, and TRAP markers

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most devastating foliar diseases of common wheat (Triticum aestivum L.) worldwide. Growing resistant cultivars is the most effective approach to control the disease. To determine inheritance of stripe rust resistance and map the resistance gene in a common wheat line D31, developed from Triticum sphaerococcum Perc. (accession number AS348), F1, F2, and BC1 progenies derived from the Taichung 29xa0×xa0D31 cross were firstly inoculated with Chinese PST race CYR32 during whole growth stages under the field conditions. Genetic analysis indicated that the resistance to CYR32 in the line D31 was conferred by one recessive gene, temporarily designated as YrSph. A total of 400 simple sequence repeat (SSR), 315 pairs of sequence-related amplified polymorphism and 42 pairs of target region amplified polymorphism markers were screened, and four SSR markers and three TRAP markers were found to be polymorphic between the resistant and susceptible DNA bulks as well as their parents. Genetic linkage was tested on segregating F2 population and indicated that all of the ten markers were linked to the resistance gene, two of which flanked the locus at 8.5 and 6.9xa0cM, respectively. The SSR markers mapped the resistance gene on chromosome arm 2AS. The results of chromosome location and pedigree analysis indicate that YrSph was probably a novel stripe rust resistance gene.

Molecular Characterization of Two Silenced y‐type Genes for Glu‐B1 in Triticum aestivum ssp. yunnanese and ssp. tibetanum

The high molecular weight glutenin subunits (HMW-GSs) are a major class of common wheat storage proteins. The bread-making quality of common wheat flour is influenced by the composition of HMW-GSs. In the present study, two unexpressed 1By genes from Triticum aesitvum L.ssp.yunnanese AS332 and T. aesitvum ssp.tibetanum AS908 were respectively cloned and characterized. The results indicated that both of the silenced 1By genes in AS332 and AS908 were 1By9. In contrast to previously reported mechanisms for silenced genes 1Ax and 1Ay, which was due to the insertion of transposon elements or the presence of premature stop codon via base substitution of C-->T transition in trinucleotides CAA or CAG, the silence of 1By9 genes was caused by premature stop codons via the deletion of base A in trinucleotide CAA, which lead to frameshift mutation and indirectly produced several premature stop codons (TAG) downstream of the coding sequence.