HaploCatcher: An R package for prediction of haplotypes

Abstract

Wheat (Triticum aestivum L.) is crucial to global food security but is often threatened by diseases, pests, and environmental stresses. Wheat-stem sawfly (Cephus cinctus Norton) poses a major threat to food security in the United States, and solid-stem varieties, which carry the stem-solidness locus (Sst1), are the main source of genetic resistance against sawfly. Marker-assisted selection uses molecular markers to identify lines possessing beneficial haplotypes, like that of the Sst1 locus. In this study, an R package titled “HaploCatcher” was developed to predict specific haplotypes of interest in genome-wide genotyped lines. A training population of 1056 lines genotyped for the Sst1 locus, known to confer stem solidness, and genome-wide markers was curated to make predictions of the Sst1 haplotypes for 292 lines from the Colorado State University wheat breeding program. Predicted Sst1 haplotypes were compared to marker-derived haplotypes. Our results indicated that the training set was substantially predictive, with kappa scores of 0.83 for k-nearest neighbors and 0.88 for random forest models. Forward validation on newly developed breeding lines demonstrated that a random forest model, trained on the total available training data, had comparable accuracy between forward and cross-validation. Estimated group means of lines classified by haplotypes from PCR-derived markers and predictive modeling did not significantly differ. The HaploCatcher package is freely available and may be utilized by breeding programs, using their own training populations, to predict haplotypes for whole-genome sequenced early generation material.

Genetic dissection of domestication traits in interspecific chickpea populations

Abstract

Chickpea (Cicer arietinum) is a pulse crop that provides an integral source of nutrition for human consumption. The close wild relatives Cicer reticulatum and Cicer echinospermum harbor untapped genetic diversity that can be exploited by chickpea breeders to improve domestic varieties. Knowledge of genomic loci that control important chickpea domestication traits will expedite the development of improved chickpea varieties derived from interspecific crosses. Therefore, we set out to identify genomic loci underlying key chickpea domestication traits by both association and quantitative trait locus (QTL) mapping using interspecific F2 populations. Diverse phenotypes were recorded for various agronomic traits. A total of 11 high-confidence markers were detected on chromosomes 1, 3, and 7 by both association and QTL mapping; these were associated with growth habit, flowering time, and seed traits. Furthermore, we identified candidate genes linked to these markers, which advanced our understanding of the genetic basis of domestication traits and validated known genes such as the FLOWERING LOCUS gene cluster that regulates flowering time. Collectively, this study has elucidated the genetic basis of chickpea domestication traits, which can facilitate the development of superior chickpea varieties.

Characterization of the CMS genetic regulation through comparative complete mitochondrial genome sequencing in Nicotiana tabacum

Abstract

Mitochondrial genomes (mitogenomes) of flowering plants vary greatly in structure and size, which can lead to frequent gene mutation, rearrangement, or recombination, then result in the cytoplasmic male sterile (CMS) mutants. In tobacco (Nicotiana tabacum), suaCMS lines are widely used in heterosis breeding; however, the related genetic regulations are not very clear. In this study, the cytological observation indicated that the pollen abortion of tobacco suaCMS(HD) occurred at the very early stage of the stamen primordia differentiation. In this study, the complete mitochondrial genomes of suaCMS(HD) and its maintainer HD were sequenced using the PacBio and Illumina Hiseq technology. The total length of the assembled mitogenomes of suaCMS(HD) and HD was 494,317 bp and 430,694 bp, respectively. Comparative analysis indicated that the expanded 64 K bases in suaCMS(HD) were mainly located in noncoding regions, and 23 and 21 big syntenic blocks (>5000 bp) were found in suaCMS(HD) and HD with a series of repeats. Electron transport chain-related genes were highly conserved in two mitogenomes, except five genes (ATP4, ATP6, COX2, CcmFC, and SDH3) with substantial substitutions. Three suaCMS(HD)-specific genes, orf261, orf291, and orf433, were screened. Sequence analysis and RT-PCR verification showed that they were unique to suaCMS(HD). Further gene location analysis and protein property prediction indicated that all the three genes were likely candidates for suaCMS(HD). This study provides new insight into understanding the suaCMS mechanism and is useful for improving tobacco breeding.

Accelerating genetic gains for quantitative resistance to verticillium wilt through predictive breeding in strawberry

Abstract

Verticillium wilt (VW), a devastating vascular wilt disease of strawberry (Fragaria × ananassa), has caused economic losses for nearly a century. This disease is caused by the soil-borne pathogen Verticillium dahliae, which occurs nearly worldwide and causes disease in numerous agriculturally important plants. The development of VW-resistant cultivars is critically important for the sustainability of strawberry production. We previously showed that a preponderance of the genetic resources (asexually propagated hybrid individuals) preserved in public germplasm collections were moderately to highly susceptible and that genetic gains for increased resistance to VW have been negligible over the last 60 years. To more fully understand the challenges associated with breeding for increased quantitative resistance to this pathogen, we developed and phenotyped a training population of hybrids (n=564$n = 564$) among elite parents with a wide range of resistance phenotypes. When these data were combined with training data from a population of elite and exotic hybrids (n=386$n = 386$), genomic prediction accuracies of 0.47–0.48 were achieved and were predicted to explain 70%–75% of the additive genetic variance for resistance. We concluded that breeding values for resistance to VW can be predicted with sufficient accuracy for effective genomic selection with routine updating of training populations.

Identification of sources of resistance to Striga gesnerioides Willd. Vatke among cowpea (Vigna unguiculata [L.] Walp) germplasm from Togo

Abstract

Striga gesnerioides is one of the major biotic constraints to cowpea (Vigna unguiculata [L.] Walp) production throughout West Africa. The best way to control and limit the spread of this weed remains genetic control, but recent works have shown that all cowpea varieties commonly grown in Togo are susceptible to S. gesnerioides. This study aims to investigate the resistance response of cowpea germplasm from Togo to S. gesnerioides. A field test was performed under natural infestation followed by a pots test under artificial infestation. Genotyping was carried out with three molecular markers SSR1, 61RM2 and C42-2B, known for their linkage to S. gesnerioides resistance genes. The results have revealed three cowpea accessions, TG20_66, TG20_82 and TG20_108, phenotypically resistant to S. gesnerioides. All three resistant accessions have shown the RSG3-301 resistance gene presence. Potential resistance gene sources to S. gesnerioides exist among cowpea accessions from Togo.

Genetic analysis and characterisation of Cmowf, a gene controlling the white petal colour phenotype in pumpkin (Cucurbita moschata D)

Abstract

Flower colour, as an important morphological marker, plays an essential role in improving the identification efficiency of the purity seed in hybrid production. However, the molecular mechanism of white-flower trait has not been reported in pumpkin (Cucurbita moschata D.). In this study, we obtained a white-flower mutant (wf) through the ethyl methane sulfonate (EMS) mutagenesis of inbred line N87 (yellow flower). F2 populations were then constructed by crossing wf mutant and N87 plant to fine map the genes controlling white-flower trait in pumpkin. Phenotypic identification revealed that carotenoid content significantly decreased in the petals of wf mutants compared with N87 plants. Genetic analysis indicated that the white flower mutant trait was controlled by a single recessive gene, Cmowf. Using bulked segregant analysis and KASP phenotyping, Cmowf was mapped to a 762 kb region on chromosome 14 containing three annotated genes. Among them, a nonsynonymous single-nucleotide polymorphisms mutation was identified only in CmoCh14G005820 gene, which encoded a DUF1997 family protein. Compared with CmoDUF1997 amino acid sequences between the wf mutants and N87 plants, the critical amino acid mutations (early termination of amino acids) occurred in wf mutants, so CmoCh14G005820 was predicted as a potential candidate for controlling the white-flower trait. RNA-sequencing analysis revealed that the expression of CmoCh14G005820 and most genes involved in carotenoid biosynthesis was significantly downregulated in wf mutants, whereas the expression of several genes responsible for carotenoid degradation was upregulated in wf mutants. This finding suggested that carotenoid metabolism may participate in the formation of flower colour in pumpkin. Overall, our results provided a theoretical basis for understanding the genetic mechanisms underlying white-flower formation in pumpkin.

QTL mapping of nitrogen use efficiency traits at the seedling and maturity stages under different nitrogen conditions in barley (Hordeum vulgare L.)

Abstract

Nitrogen (N) is an essential element for plant growth and development. The identification and utilization of N use efficiency (NUE) loci are essential for breeding high NUE cultivars. In this study, 15 NUE traits were measured in a recombinant inbred line population containing 121 lines derived from the cross between a cultivated barley (Baudin) and a wild barley (CN4027). The hydroponic culture was conducted with normal N and low N treatments in one-time frame, and field trials were conducted with N sufficiency and N deficiency treatments in two growing seasons. Twenty-two quantitative trait loci (QTLs) and four clusters were detected. Of them, the five stable QTLs Qgna.sau-3H for grain N concentration, Qtna.sau-3H for total N accumulation per plant, Qnhi.sau-3H for N harvest index, Qnutegy.sau-3H for N utilization efficiency for grain yield and Qanutedm.sau-3H.1 for N utilization efficiency for aboveground dry matter were co-located on chromosome 3H flanked by the markers bpb6282426 and bpb4786261. These two novel QTL clusters simultaneously controlled NUE traits at the seedling and maturity stages. Some genes related to NUE traits in intervals of the major QTLs were predicted. The significant relationships between NUE traits and agronomic and physiological traits were detected and discussed. In conclusion, this study uncovers the most promising genomic regions for the marker-assisted selection of NUE traits to improve NUE in barley.

New liguleless (lg2) maize stocks: Genetic resources for leaf architectural and haploid induction rate assessment studies

Abstract

Liguleless mutants produce defective ligules and auricles and, consequently, have more upright leaves than their ligulate counterparts, making them useful genetic material for plant architectural studies. Besides, owing to the recessive nature and amenability of the liguleless trait to phenotyping at the seedling stage, liguleless mutants are popularly used for ‘proof-of-concept’ demonstration and assessment of haploid induction rate (HIR) of haploid inducer lines (HILs) in maize. The commonly used liguleless testers in maize are of temperate origin and are challenging to use and maintain under tropical/sub-tropical conditions. In the present study, liguleless lines (V 601, V 602, V 603 and V 604) derived from crosses between agronomically superior locally adapted tropical ligulate lines (V 407 and CM 152) and liguleless donors of temperate origin (PDH-3 and PDH-8) were evaluated for different agro-morphological traits. Liguleless line V 602 was also used as a tester to assess the HIR of haploid inducer line EC937890 (CIM2GTAILP2). The results showed a mean HIR of 12.42% for EC937890, consistent with the HIR reported in other studies, thus demonstrating the efficacy of V 602 as a tester for determining HIR. The agronomically superior liguleless maize lines reported in this study will, therefore, be a valuable resource for leaf architectural studies, assessment of HIR of candidate HILs and maintenance of high HIR in the HILs presently in wide use in the doubled haploid (DH) programmes. Additionally, these genetic stocks carry the liguleless trait in genetic backgrounds with known heterotic affinity with early maturity Indian public maize germplasm and, therefore, can be used directly as parents in hybrid development programmes.

New Melastomataceae hosts of Chrysoporthe species in Brazil

Abstract

The angiosperm Melastomataceae family is one of the most abundant plant families worldwide and in the Brazilian cerrado, with significant environmental importance in regenerating degraded areas, especially those previously occupied by pastures. Recently, Chrysoporthe Gryzenhout & M. J. Wingf. species were reported in Brazil, causing canker, branch dieback, and mortality in native Melastomataceae. This leads to the demand for further investigation and understanding of these pathosystems. During field surveys, typical signs and symptoms associated with Chrysoporthe infection were found in Rhynchanthera grandiflora (Aubl.) D C. and Miconia theaezans (Bonpl.) Cogn. in southern Minas Gerais. Through phylogenetic analysis of the BT1 and BT2 fragments of the β-tubulin gene and morphological characterization of the isolates obtained, it was possible to identify C. doradensis Gryzenh. & M. J. Wingf. occurring in R. grandiflora and C. puriensis M. E. S. Oliv., T. P. F. Soar. & M. A. Ferr. occurring in R. grandiflora and M. theaezans. Furthermore, pathogenicity assays confirmed the pathogenicity of both species to their hosts.

Characterization of Alternaria alternata isolates from different citrus species grown in Tunisian Cap Bon peninsula

Abstract

The prospection of citrus trees affected by Alternaria alternata in Tunisian Cap Bon peninsula for 2 years, 2018 and 2019, showed a variation in the percentage of isolation frequency depending on the region, age of citrus trees and citrus species and varieties. Thirty isolates of A. alternata from citrus species were characterized and studied for their variability. The isolates were subjected to morphological identification using macroscopic and microscopic features and molecular characterization through PCR amplification of their internal transcribed spacer regions. A high morphological and molecular diversity within A. alternata isolates was detected. The molecular sequencing results precisely confirmed that these fungal isolates belong to A. alternata strains. Pathogenicity test showed that A. alternata T1 and T9 isolates were capable of causing disease symptoms only on young leaves of clementine plant (MA3 variety). These findings are useful in the development of sustainable strategies to manage Alternaria in citrus-growing areas in Tunisia.