Mingshan You

High-Density Genetic Linkage Map Construction and QTL Mapping of Grain Shape and Size in the Wheat Population Yanda1817 × Beinong6

High-density genetic linkage maps are necessary for precisely mapping quantitative trait loci (QTLs) controlling grain shape and size in wheat. By applying the Infinium iSelect 9K SNP assay, we have constructed a high-density genetic linkage map with 269 F 8 recombinant inbred lines (RILs) developed between a Chinese cornerstone wheat breeding parental line Yanda1817 and a high-yielding line Beinong6. The map contains 2431 SNPs and 128 SSR & EST-SSR markers in a total coverage of 3213.2 cM with an average interval of 1.26 cM per marker. Eighty-eight QTLs for thousand-grain weight (TGW), grain length (GL), grain width (GW) and grain thickness (GT) were detected in nine ecological environments (Beijing, Shijiazhuang and Kaifeng) during five years between 2010–2014 by inclusive composite interval mapping (ICIM) (LOD≥2.5). Among which, 17 QTLs for TGW were mapped on chromosomes 1A, 1B, 2A, 2B, 3A, 3B, 3D, 4A, 4D, 5A, 5B and 6B with phenotypic variations ranging from 2.62% to 12.08%. Four stable QTLs for TGW could be detected in five and seven environments, respectively. Thirty-two QTLs for GL were mapped on chromosomes 1B, 1D, 2A, 2B, 2D, 3B, 3D, 4A, 4B, 4D, 5A, 5B, 6B, 7A and 7B, with phenotypic variations ranging from 2.62% to 44.39%. QGl.cau-2A.2 can be detected in all the environments with the largest phenotypic variations, indicating that it is a major and stable QTL. For GW, 12 QTLs were identified with phenotypic variations range from 3.69% to 12.30%. We found 27 QTLs for GT with phenotypic variations ranged from 2.55% to 36.42%. In particular, QTL QGt.cau-5A.1 with phenotypic variations of 6.82–23.59% was detected in all the nine environments. Moreover, pleiotropic effects were detected for several QTL loci responsible for grain shape and size that could serve as target regions for fine mapping and marker assisted selection in wheat breeding programs.

Proteomic Analysis of Wheat Seed in Response to Drought Stress

Abstract Drought stress is one of the major factors affecting in wheat yield and grain quality. In order to investigate how drought stress might influence wheat quality during grain filling, three wheat cultivars Gaocheng 8901, Jagger and Nongda 3406 were subjected to drought stress during the grain filling stage. Neither globulin and glutenin, nor the relative percentage of amylose significantly changed following drought treatments, whereas albumin and gliadin concentrations did. The SDS-sedimentation, which has a strong linear correlation with wheat baking quality was markedly decreased following drought stress. These results indicated that drought had an adverse effect on wheat quality. In order to investigate the protein complexes in the wheat flour, the data from native PAGE and SDS-PAGE were combined and a total of 14 spots were successfully identified, and of these eight protein types were determined to be potential complex forming proteins.

Differential effects of a post-anthesis heat stress on wheat ( Triticum aestivum L.) grain proteome determined by iTRAQ

Heat stress, a major abiotic stressor of wheat (Triticum aestivum L.), often results in reduced yield and decreased quality. In this study, a proteomic method, Tags for Relative and Absolute Quantitation Isobaric (iTRAQ), was adopted to analyze the protein expression profile changes among wheat cultivar Jing411 under heat stress. Results indicated that there were 256 different proteins expressed in Jing411 under heat stress. According to the result of gene annotation and functional classification, 239 proteins were annotated by 856 GO function entries, including growth and metabolism proteins, energy metabolism proteins, processing and storage proteins, defense-related proteins, signal transduction, unknown function proteins and hypothetical proteins. GO enrichment analysis suggested that the differentially expressed proteins in Jing411 under heat stress were mainly involved in stimulus response (67), abiotic stress response (26) and stress response (58), kinase activity (12), and transferase activity (12). Among the differentially expressed proteins in Jing411, 115 were attributed to 119 KEGG signaling/metabolic pathways. KEGG pathway enrichment analysis in Jing411 showed that heat stress mainly affected the starch and sucrose metabolism as well as protein synthesis pathway in the endoplasmic reticulum. The protein interaction network indicated that there were 8 differentially expressed proteins that could form an interaction network in Jing411.

QTL mapping revealed TaVp-1A conferred pre-harvest sprouting resistance in wheat population Yanda 1817×Beinong 6

Abstract Pre-harvest sprouting (PHS) occurs frequently in most of the wheat cultivation area worldwide, which severely reduces yield and end-use quality, resulting in substantial economic loss. In this study, quantitative trait loci (QTL) for PHS resistance were mapped using an available high-density single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) genetic linkage map developed from a 269 recombinant inbred lines (RILs) population of Yanda 1817×Beinong 6. Using phenotypic data on two locations (Beijing and Shijiazhuang, China) in two years (2012 and 2013 harvesting seasons), five QTLs, designated as QPhs.cau-3A.1, QPhs.cau-3A.2, QPhs.cau-5B, QPhs.cau-4A, and QPhs.cau-6A, for PHS (GP) were detected by inclusive composite interval mapping (ICIM) (LOD≥2.5). Two major QTLs, QPhs.cau-3A.2 and QPhs.cau-5B, were mapped on 3AL and 5BS chromosome arms, explaining 6.29–21.65% and 4.36–5.94% of the phenotypic variance, respectively. Precise mapping and comparative genomic analysis revealed that the TaVp-1A flanking region on 3AL is responsible for QPhs.cau-3A.2. SNP markers flanking QPhs.cau-3A.2 genomic region were developed and could be used for introgression of PHS tolerance into high yielding wheat varieties through marker-assisted selection (MAS).

Transcriptome analysis of wheat grain using RNA-Seq

With the increase in consumer demand, wheat grain quality improvement has become a focus in China and worldwide. Transcriptome analysis is a powerful approach to research grain traits and elucidate their genetic regulation. In this study, two cDNA libraries from the developing grain and leaf-stem components of bread wheat cultivar, Nongda211, were sequenced using Roche/454 technology. There were 1061274 and 1516564 clean reads generated from grain and leaf-stem, respectively. A total of 61393 high-quality unigenes were obtained with an average length of 1456 bp after de novo assembly. The analysis of the 61393 unigenes involved in the biological processes of the grain showed that there were 7355 differentially expressed genes upregulated in the grain library. Gene ontology enrichment and the Kyoto Ency- clopedia of Genes and Genomes pathway enrichment analysis showed that many transcription products and transcription factors associated with carbohydrate and protein metabolism were abundantly expressed in the grain. These results contribute to excavate genes associated with wheat quality and further study how they interact.

Plant-mediated RNAi of grain aphid CHS1 gene confers common wheat resistance against aphids: RNAi of grain aphid CHS1

BACKGROUND Chitin is an important component of the insect exoskeleton and peritrophic membrane. Chitin synthase 1 (CHS1) is a key enzyme in the chitin synthesis pathway, and has a role in insect molting and growth. Plant-mediated RNA interference (RNAi) has been used as a more target-specific and environmentally safe approach to prevent and control agricultural insects. The aims of this study were to use grain aphid (Sitobion avanae) CHS1 as the target gene and to produce transgenic wheat lines for aphid control via plant-mediated RNAi. RESULTS Expression levels of CHS1 changed at different developmental stages. After feeding on the representative T3 transgenic lines Tb5-2 and Tb10-3, CHS1 expression levels in grain aphid decreased by 50.29% and 45.32%, respectively; and total and molting aphid numbers reduced significantly, compared with controls. Consistent with this, aphid numbers in mixed natural populations reduced significantly in the respective T4 and T5 transgenic lines under field conditions, and T5 transgenic lines had higher grain weight compared with the unsprayed insecticide wild-type and insecticide-sprayed wild-type. CONCLUSION These results indicate that plant-mediated RNAi of the grain aphid CHS1 gene confers common wheat resistance against aphids.