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Integrating high-throughput phenotyping (HTP) based traits into phenomic and genomic selection (GS) can accelerate the breeding of high-yielding and climate-resilient wheat cultivars. In this study, we explored the applicability of Unmanned Aerial Vehicles (UAV)-assisted HTP combined with deep learning (DL) for the phenomic or multi-trait (MT) genomic prediction of grain yield (GY), test weight (TW), and grain protein content (GPC) in winter wheat. Significant correlations were observed between agronomic traits and HTP-based traits across different growth stages of winter wheat. Using a deep neural network (DNN) model, HTP-based phenomic predictions showed robust prediction accuracies for GY, TW, and GPC for a single location with R2 of 0.71, 0.62, and 0.49, respectively. Further prediction accuracies increased (R2 of 0.76, 0.64, and 0.75) for GY, TW, and GPC, respectively when advanced breeding lines from multi-locations were used in the DNN model. Prediction accuracies for GY varied across growth stages, with the highest accuracy at the Feekes 11 (Milky ripe) stage. Furthermore, forward prediction of GY in preliminary breeding lines using DNN trained on multi-location data from advanced breeding lines improved the prediction accuracy by 32% compared to single-location data. Next, we evaluated the potential of incorporating HTP-based traits in multi-trait genomic selection (MT-GS) models in the prediction of GY, TW, and GPC. MTGS, models including UAV data-based anthocyanin reflectance index (ARI), green chlorophyll index (GCI), and ratio vegetation index 2 (RVI_2) as covariates demonstrated higher predictive ability (0.40, 0.40, and 0.37, respectively) as compared to single-trait model (0.23) for GY. Overall, this study demonstrates the potential of integrating HTP traits into DL-based phenomic or MT-GS models for enhancing breeding efficiency. [ABSTRACT FROM AUTHOR]

In recent years, stripe rust has become an increasingly problematic disease affecting wheat (Triticum aestivum L.). Although stripe rust resistance genes Yr28 and YrAS2388R have been successfully transferred from Aegilops tauschii Coss. to hexaploid wheat, these genes have not been widely used for stripe rust resistance breeding largely due to a lack of publicly available and well‐adapted germplasm. Here we describe U6719‐004 (Entry 538 of D‐genome nested association mapping population, Reg. No. MP‐14, NSL 536301 MAP), a hexaploid wheat germplasm carrying YrAS2388R stripe rust resistance derived from Ae. tauschii accession TA1718 that is well adapted to U.S. wheat production environments. U6719‐004 (KS05HW14‐3/3/KS05HW14‐3/TA1718//KS05HW14‐3) was developed using a combination of direct hybridization of TA1718 with the hard white winter wheat line KS05HW14‐3 followed by two rounds of backcrossing and three generations of single seed descent. U6719‐004 stripe rust resistance was characterized under controlled and natural conditions. Using gene‐based markers, TA1718 and U6719‐004 were confirmed to be YrAS2388R carriers. Grain yield for U6719‐004 was evaluated in 11 year‐by‐location environments across the United States. Grain quality parameters were measured in four year‐by‐location environments. Compared with the recurrent wheat parent KS05HW14‐3 and locally adapted check cultivars, the Ae. tauschii introgression had little to no impact on U6719‐004 grain yield or quality. The availability of U6719‐004 will enable the use of YrAS2388R for stripe rust resistance breeding efforts. Core Ideas: The hexaploid wheat germplasm U6719‐004 carries stripe rust resistance gene YrAS2388R.U6719‐004 is broadly adapted to wheat production environments of the United States.U6719‐004 has acceptable grain yield and grain quality. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB), also known as scab, is a devastating fungal disease of wheat that causes significant losses in grain yield and quality. Quantitative inheritance and cumbersome phenotyping make FHB resistance a challenging trait for direct selection in wheat breeding. Genomic selection to predict FHB resistance traits has shown promise in several studies. Here, we used univariate and multivariate genomic prediction models to evaluate the prediction accuracy (PA) for different FHB traits using 476 elite and advanced breeding lines developed by South Dakota State University hard winter wheat breeding program. These breeding lines were assessed for FHB disease index (DIS), and percentage of Fusarium damaged kernels (FDK) in three FHB nurseries in 2018, 2019, and 2020 (TP18, TP19, and TP20) and were evaluated as training populations (TP) for genomic prediction (GP) of FHB traits. We observed a moderate PA using univariate models for DIS (0.39 and 0.35) and FDK (0.35 and 0.37) using TP19 and TP20, respectively, while slightly higher PA was observed (0.41 for DIS and 0.38 for FDK) when TP19 and TP20 (TP19 + 20) were combined to leverage the advantage of a large training population. Although GP with multivariate approach including plant height and days to heading as covariates did not significantly improve PA for DIS and FDK over univariate models, PA for DON increased by 20% using DIS, FDK, DTH as covariates using multi-trait model in 2020. Finally, we used TP19, TP20, and TP19 + 20 in forward prediction to calculate genomic-estimated breeding values (GEBVs) for DIS and FDK in preliminary breeding lines at an early stage of the breeding program. We observed moderate PA of up to 0.59 for DIS and 0.54 for FDK, demonstrating the promise in genomic prediction for FHB resistance in earlier stages using advanced lines. Our results suggest GP for expensive FHB traits like DON and FDK can facilitate the rejection of highly susceptible materials at an early stage in a breeding program. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB), caused by the fungus Fusarium graminearum Schwabe is an important disease of wheat that causes severe yield losses along with serious quality concerns. Incorporating the host resistance from either wild relatives, landraces, or exotic materials remains challenging and has shown limited success. Therefore, a better understanding of the genetic basis of native FHB resistance in hard winter wheat (HWW) and combining it with major quantitative trait loci (QTLs) can facilitate the development of FHB-resistant cultivars. In this study, we evaluated a set of 257 breeding lines from the South Dakota State University (SDSU) breeding program to uncover the genetic basis of native FHB resistance in the US hard winter wheat. We conducted a multi-locus genomewide association study (ML-GWAS) with 9,321 high-quality single-nucleotide polymorphisms (SNPs). A total of six distinct marker-trait associations (MTAs) were identified for the FHB disease index (DIS) on five different chromosomes including 2A, 2B, 3B, 4B, and 7A. Further, eight MTAs were identified for Fusarium-damaged kernels (FDK) on six chromosomes including 3B, 5A, 6B, 6D, 7A, and 7B. Out of the 14 significant MTAs, 10 were found in the proximity of previously reported regions for FHB resistance in different wheat classes and were validated in HWW, while four MTAs represent likely novel loci for FHB resistance. Accumulation of favorable alleles of reported MTAs resulted in significantly lower mean DIS and FDK score, demonstrating the additive effect of FHB resistance alleles. Candidate gene analysis for two important MTAs identified several genes with putative proteins of interest; however, further investigation of these regions is needed to identify genes conferring FHB resistance. The current study sheds light on the genetic basis of native FHB resistance in the US HWW germplasm and the resistant lines and MTAs identified in this study will be useful resources for FHB resistance breeding via marker-assisted selection. [ABSTRACT FROM AUTHOR]

Powdery mildew of wheat is a foliar disease that is spread worldwide. Cultivation of resistant varieties is the most effective, economical, and environmentally friendly strategy to curb this disease. Powdery mildew resistance genes (Pm) are the primary resources for resistance breeding, and new Pm genes are in constant demand. Previously, we identified Aegilops longissima chromosome 6Sl#3 as a carrier of powdery mildew resistance and designated the resistance gene as Pm6Sl. Here, we reported the design of 24 markers specific to 6Sl#3 on the basis of the full-length cDNA sequences of 6Sl#3 donor Ae. longissma accession TA1910, and the development of wheat- Ae. longissima 6Sl#3 introgression stocks by ph1b- induced homoeologous recombination. Further, 6Sl#3 introgression lines were identified and characterized by integration analysis of powdery mildew responses, in situ hybridization, and molecular markers and Pm6Sl was mapped to a distal interval of 42.80 Mb between markers Ael58410 and Ael57699 in the long arm of 6Sl#3. Two resistant recombinants, R43 (T6BS.6BL-6Sl#3L) and T27 (Ti6AS.6AL-6Sl#3L-6AL), contained segments harboring Pm6Sl with less than 8% of 6Sl#3 genomic length, and two markers were diagnostic for Pm6Sl. This study broadened powdery mildew resistance gene resources for wheat improvement and provided a fundamental basis for fine mapping and cloning of Pm6Sl to further understand its molecular mechanism of disease resistance. [ABSTRACT FROM AUTHOR]

Abstract Background Triticum monococcum L., an A genome diploid einkorn wheat, was the first domesticated crop. As a diploid, it is attractive genetic model for the study of gene structure and function of wheat-specific traits. Diploid wheat is currently not amenable to reverse genetics approaches such as insertion mutagenesis and post-transcriptional gene silencing strategies. However, TILLING offers a powerful functional genetics approach for wheat gene analysis. Results We developed a TILLING population of 1,532 M2 families using EMS as a mutagen. A total of 67 mutants were obtained for the four genes studied. Waxy gene mutation frequencies are known to be 1/17.6 - 34.4 kb DNA in polyploid wheat TILLING populations. The T. monococcum diploid wheat TILLING population had a mutation frequency of 1/90 kb for the same gene. Lignin biosynthesis pathway genes- COMT1, HCT2, and 4CL1 had mutation frequencies of 1/86 kb, 1/92 kb and 1/100 kb, respectively. The overall mutation frequency of the diploid wheat TILLING population was 1/92 kb. Conclusion The mutation frequency of a diploid wheat TILLING population was found to be higher than that reported for other diploid grasses. The rate, however, is lower than tetraploid and hexaploid wheat TILLING populations because of the higher tolerance of polyploids to mutations. Unlike polyploid wheat, most mutants in diploid wheat have a phenotype amenable to forward and reverse genetic analysis and establish diploid wheat as an attractive model to study gene function in wheat. We estimate that a TILLING population of 5, 520 will be needed to get a non-sense mutation for every wheat gene of interest with 95% probability.

Abstract Background Bread wheat, one of the world’s staple food crops, has the largest, highly repetitive and polyploid genome among the cereal crops. The wheat genome holds the key to crop genetic improvement against challenges such as climate change, environmental degradation, and water scarcity. To unravel the complex wheat genome, the International Wheat Genome Sequencing Consortium (IWGSC) is pursuing a chromosome- and chromosome arm-based approach to physical mapping and sequencing. Here we report on the use of a BAC library made from flow-sorted telosomic chromosome 3A short arm (t3AS) for marker development and analysis of sequence composition and comparative evolution of homoeologous genomes of hexaploid wheat. Results The end-sequencing of 9,984 random BACs from a chromosome arm 3AS-specific library (TaaCsp3AShA) generated 11,014,359 bp of high quality sequence from 17,591 BAC-ends with an average length of 626 bp. The sequence represents 3.2% of t3AS with an average DNA sequence read every 19 kb. Overall, 79% of the sequence consisted of repetitive elements, 1.38% as coding regions (estimated 2,850 genes) and another 19% of unknown origin. Comparative sequence analysis suggested that 70-77% of the genes present in both 3A and 3B were syntenic with model species. Among the transposable elements, gypsy/sabrina (12.4%) was the most abundant repeat and was significantly more frequent in 3A compared to homoeologous chromosome 3B. Twenty novel repetitive sequences were also identified using de novo repeat identification. BESs were screened to identify simple sequence repeats (SSR) and transposable element junctions. A total of 1,057 SSRs were identified with a density of one per 10.4 kb, and 7,928 junctions between transposable elements (TE) and other sequences were identified with a density of one per 1.39 kb. With the objective of enhancing the marker density of chromosome 3AS, oligonucleotide primers were successfully designed from 758 SSRs and 695 Insertion Site Based Polymorphisms (ISBPs). Of the 96 ISBP primer pairs tested, 28 (29%) were 3A-specific and compared to 17 (18%) for 96 SSRs. Conclusion This work reports on the use of wheat chromosome arm 3AS-specific BAC library for the targeted generation of sequence data from a particular region of the huge genome of wheat. A large quantity of sequences were generated from the A genome of hexaploid wheat for comparative genome analysis with homoeologous B and D genomes and other model grass genomes. Hundreds of molecular markers were developed from the 3AS arm-specific sequences; these and other sequences will be useful in gene discovery and physical mapping.

Abstract Background Bald cypress (Taxodium distichum var. distichum) is a coniferous tree of tremendous ecological and economic importance. It is a member of the family Cupressaceae which also includes cypresses, redwoods, sequoias, thujas, and junipers. While the bald cypress genome is more than three times the size of the human genome, its 1C DNA content is amongst the smallest of any conifer. To learn more about the genome of bald cypress and gain insight into the evolution of Cupressaceae genomes, we performed a Cot analysis and used Cot filtration to study Taxodium DNA. Additionally, we constructed a 6.7 genome-equivalent BAC library that we screened with known Taxodium genes and select repeats. Results The bald cypress genome is composed of 90% repetitive DNA with most sequences being found in low to mid copy numbers. The most abundant repeats are found in fewer than 25,000 copies per genome. Approximately 7.4% of the genome is single/low-copy DNA (i.e., sequences found in 1 to 5 copies). Sequencing of highly repetitive Cot clones indicates that most Taxodium repeats are highly diverged from previously characterized plant repeat sequences. The bald cypress BAC library consists of 606,336 clones (average insert size of 113 kb) and collectively provides 6.7-fold genome equivalent coverage of the bald cypress genome. Macroarray screening with known genes produced, on average, about 1.5 positive clones per probe per genome-equivalent. Library screening with Cot-1 DNA revealed that approximately 83% of BAC clones contain repetitive sequences iterated 103 to 104 times per genome. Conclusions The BAC library for bald cypress is the first to be generated for a conifer species outside of the family Pinaceae. The Taxodium BAC library was shown to be useful in gene isolation and genome characterization and should be an important tool in gymnosperm comparative genomics, physical mapping, genome sequencing, and gene/polymorphism discovery. The single/low-copy (SL) component of bald cypress is 4.6 times the size of the Arabidopsis genome. As suggested for other gymnosperms, the large amount of SL DNA in Taxodium is likely the result of divergence among ancient repeat copies and gene/pseudogene duplication.

Preharvest sprouting ( PHS) is one of the major constraints of wheat production in areas where prolonged rainfall occurs during harvest. Ta PHS1 is a gene that regulates PHS resistance on chromosome 3A of wheat, and two causal mutations in the positions +646 and +666 of the Ta PHS1 coding region result in wheat PHS susceptibility. Three competitive allele-specific PCR ( KASP) markers were developed based on the two mutations in the coding region and one in the promoter region and validated in 82 wheat cultivars with known genotypes. These markers can be used to transfer Ta PHS1 in breeding through marker-assisted selection., Screening of 327 accessions of wheat A genome progenitors using the three KASP markers identified different haplotypes in both diploid and tetraploid wheats. Only one Triticum monococcum accession, however, carries both causal mutations in the Ta PHS1 coding region and shows PHS susceptibility. Five of 249 common wheat landraces collected from the Fertile Crescent and surrounding areas carried the mutation (C) in the promoter (−222), and one landrace carries both the causal mutations in the Ta PHS1 coding region, indicating that the mis-splicing (+646) mutation occurred during common wheat domestication., PHS assay of wheat progenitor accessions demonstrated that the wild-types were highly PHS-resistant, whereas the domesticated type showed increased PHS susceptibility., The mis-splicing Ta PHS1 mutation for PHS susceptibility was involved in wheat domestication and might arise independently between T. monococcum and Triticum aestivum. [ABSTRACT FROM AUTHOR]