Yinguang Bao

Conditional QTL mapping for plant height with respect to the length of the spike and internode in two mapping populations of wheat.

Plant height (PH) in wheat is a complex trait; its components include spike length (SL) and internode lengths. To precisely analyze the factors affecting PH, two F8:9 recombinant inbred line (RIL) populations comprising 485 and 229 lines were generated. Crosses were performed between Weimai 8 and Jimai 20 (WJ) and between Weimai 8 and Yannong 19 (WY). Possible genetic relationships between PH and PH components (PHC) were evaluated at the quantitative trait locus (QTL) level. PH and PHC (including SL and internode lengths from the first to the fourth counted from the top, abbreviated as FIITL, SITL, TITL, and FOITL, respectively) were measured in four environments. Individual and the pooled values from four trials were used in the present analysis. A QTL for PH was mapped using data on PH and on PH conditioned by PHC using IciMapping V2.2. All 21 chromosomes in wheat were shown to harbor factors affecting PH in two populations, by both conditional and unconditional QTL mapping methods. At least 11 pairwise congruent QTL were identified in the two populations. In total, ten unconditional QTL and five conditional QTL that could be detected in the conditional analysis only have been verified in no less than three trials in WJ and WY. In addition, three QTL on the short arms of chromosomes 4B, 4D, and 7B were mapped to positions similar to those of the semi-dwarfing genes Rht-B1, Rht-D1 and Rht13, respectively. Conditional QTL mapping analysis in WJ and WY proved that, at the QTL level, SL contributed the least to PH, followed by FIITL; TITL had the strongest influence on PH, followed by SITL and FOITL. The results above indicated that the conditional QTL mapping method can be used to evaluate possible genetic relationships between PH and PHC, and it can efficiently and precisely reveal counteracting QTL, which will enhance the understanding of the genetic basis of PH in wheat. The combination of two related populations with a large/moderate population size made the results authentic and accurate.

Conditional and unconditional QTL mapping of drought-tolerance-related traits of wheat seedling using two related RIL populations

For discovering the quantitative trait loci (QTLs) contributing to early seedling growth and drought tolerance during germination, conditional and unconditional analyses of 12 traits of wheat seedlings: coleoptile length, seedling height, longest root length, root number, seedling fresh weight, stem and leaves fresh weight, root fresh weight, seedling dry weight, stem and leaves dry weight, root dry weight, root to shoot fresh weight ratio, root-to-shoot dry weight ratio, were conducted under two water conditions using two F8:9 recombinant inbred line (RIL) populations. The results of unconditional analysis are as follows: 88 QTLs accounting for 3.33–77.01% of the phenotypic variations were detected on chromosomes 1A, 1B, 1D, 2A, 2B, 2D, 3A, 3B, 4A, 4B, 4D, 5A, 5B, 5D, 6A, 6B, 6D, 7A, 7B and 7D. Among these QTLs, 19 were main-effect QTLs with a contribution rate greater than 10%. The results of the conditional QTL analysis of 12 traits under osmotic stress on normal water conditions were as follows: altogether 22 QTLs concerned with drought tolerance were detected on chromosomes 1B, 2A, 2B, 3B, 4A, 5D, 6A, 6D, 7B, and 7D. Of these QTLs, six were main-effect QTLs. These 22 QTLs were all special loci directly concerned with drought tolerance and most of them could not be detected by unconditional analysis. The finding of these QTLs has an important significance for fine-mapping technique, map-based cloning, and molecular marker-assisted selection of early seedling traits, such as growth and drought tolerance.

Molecular cytogenetic identification of a novel dwarf wheat line with introgressed Thinopyrum ponticum chromatin

Novel dwarfing germplasms and dwarfing genes are valuable for the wheat breeding. A novel semi-dwarf line, 31505-1, with reduced height compared with its common wheat parent, was derived from a cross between common wheat and Thinopyrum ponticum. Cytological studies demonstrated that 31505-1 contained 42 chromosomes and formed 21 bivalents at meiotic metaphase I. Genomic in situ hybridization (GISH) analysis showed that 31505-1 had no large Th. ponticum chromosome fragments. Fluorescence in situ hybridization (FISH) results revealed the absence of a pAs1 hybridization band on 2DL chromosome of 31505-1. Two SSR markers (Xwmc41 and Xcfd168) and two STS markers (Xmag4059 and Xmag3596), which were located on 2D chromosome, amplified unique bands of Th. Ponticum in 31505-1. These revealed presence of an introgressed Th. ponticum segment in 2DL chromosome of dwarf line 31505-1, although the alien segment could not be detected by GISH.

Kernel weight per spike: what contributes to it at the individual QTL level?

Spike length (SL), spikelet number (SPN) per spike, kernel number per spike (KNPS), and thousand-kernel weight (TKW) have strong genetic associations with kernel weight per spike (KWPS) in wheat. To investigate their genetic relationships at the individual quantitative trait locus (QTL) level, both unconditional and conditional QTL mapping for KWPS with respect to SL, SPN, KNPS, and TKW were conducted. Two related F8:9 recombinant inbred line populations, comprising 485 and 229 lines, respectively, were used. The trait phenotypic performances of each population were evaluated in four different environments. Unconditional QTL mapping analysis identified 22 putative additive QTL for KWPS, five of which were stable QTL, and only QKwps-WJ-1B.2 showed significant additive-by-environment interaction effects. In comparison with unconditional QTL mapping analysis, conditional QTL mapping analysis indicated that, at the QTL level, KNPS and TKW contributed more to KWPS than did SL and SPN. Any unconditional QTL for KWPS detected in this study was associated with at least one of its four related traits. The present study will provide assistance in the understanding of the genetic relationships between KWPS and its related traits.

Molecular cytogenetic characterization of a new wheat-Thinopyrum intermedium partial amphiploid resistant to powdery mildew and stripe rust.

A partial amphiploid, TE253, derived from crosses between the common wheat (Triticum aestivum L.) cultivar Yannong 15 and Thinopyrum intermedium (Host) Barkworth & D.R. Dewey was characterized by cytogenetic observations, disease resistance tests and genomic in situ hybridization (GISH). Mitotic observations showed that most plants of TE253 had 56 chromosomes, but a few had 54 or 55 chromosomes. The chromosomes in most pollen mother cells of plants with 2n = 56 formed 28 bivalents. Univalents (0.89 per cell) and tetravalents (0.087 per cell) occasionally occurred at meiotic metaphase I, showing a high degree of cytogenetic stability. After inoculation with the powdery mildew and stripe rust pathogens, Yannong 15 was highly susceptible, whereas TE253 and Th. intermedium were immune to both diseases. This indicated that the resistance of TE253 to powdery mildew and stripe rust was derived from Th. intermedium. GISH analysis using St-genomic DNA from Pseudoroegneria strigosa (M. Bieb) Á. Löve as a probe and ABD-genomic DNA from Chinese Spring wheat as a blocker demonstrated that TE253 consisted of 2 St-genome chromosomes, 8 JS-genome chromosomes, 2 SAT J chromosomes and 2 J-St translocated chromosomes. Line TE253 is a new partial amphiploid with resistance to both powdery mildew and stripe rust and can be used as a source of resistance genes in wheat improvement.

QTL detection of seven quality traits in wheat using two related recombinant inbred line populations

Grain protein content (GPC) and gluten quality are the most important factors determining the end-use quality of wheat for pasta-making. Both GPC and gluten quality are considered to be polygenic traits influenced by environmental factors and other agricultural practices. Two related F8:9 recombinant inbred line (RIL) populations were generated to localise genetic factors controlling seven quality traits: GPC, wet gluten content (WGC), flour whiteness (FW), kernel hardness (KH), water absorption (Abs), dough development time (DDT) and dough stability time (DST). These lines were derived by crossing Weimai 8 and Jimai 20 (WJ) and by crossing Weimai 8 and Yannong 19 (WY). In total, WJ comprised 485 lines, while WY comprised 229 lines. Data on these seven quality traits were collected from each line in five different environments. Up to 85 putative QTLs for the seven traits were detected in WJ and 65 putative QTLs were detected in WY. Of these QTLs, 31 QTLs (36.47%) were detected in at least two trials in WJ, while 24 QTLs (36.92%) were detected in at least two trials in WY. Three QTLs from WJ and 25 from WY accounted for more than 10% of the phenotypic variance. The total 150 QTLs were spread throughout all 21 wheat chromosomes. Of these, at least thirteen pairwise were common to both populations, accounting for 20.00 and 15.29% of the total QTLs in WJ and WY, respectively. A major QTL for GPC, accounting for 53.04% of the phenotypic variation, was detected on chromosome 5A. A major QTL for WGC also shared this interval, explained more than 36% of the phenotypic variation, and was significant in two environments. Though co-located QTLs were common, every trait had its unique control mechanism, even for two closely related traits. Due to the different sizes of the two line populations, we also assessed the effects of population size on the efficiency and precision of QTL detection. In sum, this study will enhance our understanding of the genetic basis of these seven pivotal quality traits and facilitate the breeding of improved wheat varieties.

Transmission of the Chromosome 1R in Winter Wheat Germplasm Aimengniu and Its Derivatives Revealed by Molecular Markers

In order to clarify the transmission of the rye chromosome 1R in winter wheat germplasm Aimengniu and its derivatives, 17 derivatives and 7 types of Aimengniu were examined through molecular-marker technology. The results showed that the chromosome arm 1RS of Neuzucht was transmitted to 5 of the 7 types of Aimengniu, i.e., Aimengniu II and Aimengniu IV-Aimengniu VII, no segment of 1RS was identified in Aimengniu I or Aimengniu III. As for the 17 derivatives, the 1RS chromosome arm of Aimengniu was transmitted to 11 derivatives, part segments of 1RS were found in 1 derivative, while no segment was found in the remaining 5 ones. The results provided the evidence that molecular-marker technology was an efficient approach and suitable for analysis of the transmission of chromosome 1R.

Molecular cytogenetic identification of a wheat (Triticum aestivum)-American dune grass (Leymus mollis) translocation line resistant to stripe rust.

Leymus mollis, a perennial allotetraploid (2n = 4x = 28), known as American dune grass, is a wild relative of wheat that could be useful for cultivar improvement. Shannong0096, developed from interspecific hybridization between common wheat cv. Yannong15 and L. mollis, was analyzed with cytological procedures, genomic in situ hybridization, stripe-rust resistance screening and molecular marker analysis. We found that Shannong0096 has 42 chromosomes in the root-tip cells at mitotic metaphase and 21 bivalents in the pollen mother cells at meiotic metaphase I, demonstrating cytogenetic stability. Genomic in situ hybridization probed with total genomic DNA from L. mollis gave strong hybridization signals in the distal region of two wheat chromosome arms. A single dominant Yr gene, derived from L. mollis and temporarily designated as YrSn0096, was found on the long arm of chromosome 4A of Shannong0096. YrSn0096 should be a novel Yr gene because none of the previously reported Yr genes on chromosome 4A are related to L. mollis. This gene was found to be closely linked to the loci Xbarc236 and Xksum134 with genetic distances of 5.0 and 4.8 cM, respectively. Based on data from 267 F(2) plants of Yannong15/Huixianhong, the linkage map of YrSn0096, using the two molecular markers, was established in the order Xbarc236-YrSn0096-Xksum134. Shannong0096 appeared to be a unique wheat-L. mollis translocation with cryptic alien introgression. Cytogenetic stability, a high level of stripe-rust resistance, the common wheat background, and other positive agronomic traits make it a desirable donor for introducing novel alien resistance genes in wheat breeding programs, with the advantage of molecular markers that can be used to confirm introgression.

Inhibitory effect of dexamethasone on Lewis mice lung cancer cells.

Recent studies have found that glucocorticoids are closely associated with oncogenesis and the development of many types of tumors. The aim of this study was to observe the effect of dexamethasone on the growth and angiogenesis of transplanted Lewis lung carcinoma in mice. Lewis lung carcinoma cells were inoculated subcutaneously into the right axilla of C57BL/6 mice, and the mice were randomly divided into 3 groups: the control group, cisplatin group, and dexamethasone group. From day 7 after inoculation, all the mice were given different treatments for 10 days, and changes in xenograft tumor volumes were monitored. All mice were sacrificed on day 17, and the tumors were obtained and weighed and the tumor inhibitory rate was calculated. The expression levels of hypoxia inducible factor 1α (HIF-1α) and vascular endothelial growth factor (VEGF), as well as the microvessel density (MVD) in the tumor mass, were measured by immunohistochemistry. Tumor growth was suppressed in the cisplatin group and dexamethasone group. The weights of tumors were markedly decreased in the cisplatin group and dexamethasone group compared with the control group (P < 0.05). The expression levels of HIF-1α and VEGF and the MVD were significantly lower in the cisplatin group and dexamethasone group than in the control group (P < 0.05). However, these levels were not significantly different between the cisplatin group and dexamethasone group (P > 0.05). Dexamethasone can effectively inhibit the growth and angiogenesis of Lewis lung carcinoma by inhibiting the expression of HIF-1α and VEGF.

Comparative Transcriptome Analyses of Resistant and Susceptible Near-Isogenic Wheat Lines following Inoculation with Blumeria graminis f. sp. tritici.

Powdery mildew is one of the most important diseases of wheat. In this study, the leaf RNA samples of wheat NILs carrying powdery mildew resistant and susceptible Pm2 alleles (L031 and Chancellor) and its F1 hybrid at two time points (16 h and 96 h postinoculation) were used for RNA-seq analysis. We carry comparison between similar materials at different times and between different materials at same times. The overlapping DEGs between the dominant phenotypes (L031 and F1 hybrid) and the recessive phenotype (Chancellor) were 1028 and 2214 DEGs, which were clearly lower than those between the dominant and recessive parents and thus could provide relatively accurate and valuable information. GO and KEGG enrichment analysis of DEGs revealed that other than the expected defense-related genes, differential up- and downregulation of genes from many other signaling networks were also involved. Comparative transcriptome analysis also revealed that early-stage postinoculation is important and suitable time points to study expression profiles and signaling pathways of resistance-related genes following fungal inoculation. qRT-PCR analyses showed highly consistent expression patterns of genes with RNA-seq data. The results will aid in the identification of genes and signaling pathways involved in powdery mildew response in wheat.