Backcross Populations Research Articles

Inheritance of aluminium tolerance in sunflower (Helianthus annuus L.)

In acidic soil, aluminium (Al) toxicity is one of the foremost restraining factors which inhibit crop performance. Breeding for Al tolerance seems to be a vital approach for enhancing productivity of field crops including sunflower in acidic soil regions. In order to understand the inheritance pattern, tolerant genotypes NDLR-06 and EC601861 were crossed with susceptible genotypes MR-06 and RHA-298. P1, P2, F1, F2 and backcross populations were raised in nutrient solution and classified as tolerant/susceptible by observing root re-growth after staining with eriochrome cyanine R. Dominance nature of Al tolerance over susceptible was confirmed by the responses of F1’s which were similar to the tolerant parents. The segregation ratio for Al tolerance vs. susceptible in the F2 and backcross generations were 3:1 and 1:1, respectively. These outcomes indicated that inheritance of Al tolerance is governed by single dominant gene which can be transferred easily to desirable genotypes via backcross breedingmethod. This is the first report on inheritance of Al tolerance in sunflower.

Mapping of Rf20(t), a minor fertility restorer gene for rice wild abortive cytoplasmic male sterility in the maintainer line 'Zhenshan97B'.

Three-line hybrid rice has primarily been developed on wild abortive (WA)-type cytoplasmic male sterility (CMS) and has helped increase the yield of rice globally. The development of WA-type CMS lines and hybrids was expedited through the identification and mapping of the fertility restorer gene (Rf) in maintainers. This study observed fertile plants in WA-TianfengA/Zhenshan97B//TianfengB population, indicating that the maintainer line 'Zhenshan97B' should carry Rfs for WA-type CMS. Several advanced backcross populations were generated with the genetic background of the 'WA-TianfengA,' and the pollen fertility levels of the backcrossed individuals in BC3F1, BC4F1 and BC4F2 populations are governed by a new gene, Rf20(t), from 'Zhenshan97B.' Employing bulk segregant analysis of fertile and sterile pools from the BC4F1 population, Rf20(t) was genetically mapped to a candidate region on chromosome 10. Subsequently, Rf20(t) was located between RM24883 and RM24919 through recombination analysis of molecular markers using the BC4F2 population. Implementing a substitution mapping strategy, Rf20(t) was ultimately mapped to a 245-kb region between the molecular markers STS10-122 and STS10-126 and obtained the most likely candidate gene LOC_Os10g02650, which is predicted to encode pentatricopeptide repeat-containing (PPR) protein. These results enhance our understanding of the fertility restoration of WA-type CMS lines, facilitating the development of high-quality pairs of WA-type CMS and maintainer lines.

Identification and Functional Characterization of Two Major Loci Associated with Resistance against Brown Planthoppers (Nilaparvata lugens (Stål)) Derived from Oryza nivara.

The brown planthopper (BPH) is a highly destructive pest of rice, causing significant economic losses in various regions of South and Southeast Asia. Researchers have made promising strides in developing resistance against BPH in rice. Introgression line RPBio4918-230S, derived from Oryza nivara, has shown consistent resistance to BPH at both the seedling and adult stages of rice plants. Segregation analysis has revealed that this resistance is governed by two recessive loci, known as bph39(t) and bph40(t), contributing to 21% and 22% of the phenotypic variance, respectively. We later mapped the genes using a backcross population derived from a cross between Swarna and RPBio4918-230S. We identified specific marker loci, namely RM8213, RM5953, and R4M17, on chromosome 4, flanking the bph39(t) and bph40(t) loci. Furthermore, quantitative expression analysis of candidate genes situated between the RM8213 and R4M17 markers was conducted. It was observed that eight genes exhibited up-regulation in RPBio4918-230S and down-regulation in Swarna after BPH infestation. One gene of particular interest, a serine/threonine-protein kinase receptor (STPKR), showed significant up-regulation in RPBio4918-230S. In-depth sequencing of the susceptible and resistant alleles of STPKR from Swarna and RPBio4918-230S, respectively, revealed numerous single nucleotide polymorphisms (SNPs) and insertion-deletion (InDel) mutations, both in the coding and regulatory regions of the gene. Notably, six of these mutations resulted in amino acid substitutions in the coding region of STPKR (R5K, I38L, S120N, T319A, T320S, and F348S) when compared to Swarna and the reference sequence of Nipponbare. Further validation of these mutations in a set of highly resistant and susceptible backcross inbred lines confirmed the candidacy of the STPKR gene with respect to BPH resistance controlled by bph39(t) and bph40(t). Functional markers specific for STPKR have been developed and validated and can be used for accelerated transfer of the resistant locus to elite rice cultivars.

Correlation and variability analysis for yield and related traits of sweet corn in backcross populations

The 14 biometrical traits of sweet corn were studied in BC2F2 and BC2F3 generations of SCM-Se-Y-1 x UMI 1230β+ to analyse the mean performance and frequency distribution patterns to select potential individuals with high yielding agronomic traits. The mean, GCV, PCV, heritability, and GAM were calculated for all the recorded traits. The Pearson correlation coefficient revealed the strong and positive association of cob weight with cob length, number of kernels in a row and number of kernel rows per cob. Similarly, single plant yield shows positive correlation with number of kernels per row, number of kernel rows per cob, cob length and cob weight. The populations exhibit high PCV than the GCV, which reflects the direct influence of environments on trait performance. The BC2F2 generation exhibits greater trait variability, while BC2F3 shows signs of stabilization. Both generations display high heritability, indicating strong genetic influences for yield related traits. Heritability and GAM for the trait cob weight and single plant yield were higher, which gives added advantage for isolating the superior individual. The results of this study suggest that the selection of agronomical traits to enhance yield in sweet corn is of utmost importance due to its far-reaching economic, nutritional, and environmental implications.

Wild barley genomic resources for drought adaptability and quality improvement

Wild species of crop plants may contain genes for adaptation to environmental stresses. Wild barley, with higher yield stability and better adaptation to climate change, is a valuable novel gene source for improving drought tolerance, grain quality, and commercial applications. In this research, nested backcross populations (NBP) were developed by crossing 21 wild barley accessions to the Ryhan03 cultivar, and the F1 progenies were backcrossed to Ryhan03 once. Screening of the 443 NBP developed lines was performed based on stress tolerance score (STS) and grain yield during two BC1F3 and BC1F4 generations, then 95 drought-tolerant and susceptible lines were selected for further evaluation in the BC1F5 generation under two water conditions. Efficient NBP strategy might be valuable for developing improved barley cultivars for nutritional quality and drought tolerance. Reasonable diversity among the BC1F5 families demonstrated that strict selection for beta-glucan content, drought tolerance, and yield is feasible and successful. The considerable variation observed for quality-related traits was significant for achieving the desired advanced barley lines to meet various consumption preferences. Wild donors contributed several favorable alleles to quality traits and drought tolerance. Progeny lines T42, T5, T60, and T35 were introduced as high-yielding, drought-tolerant lines with high beta-glucan content.

Genetic analysis of basal stalk rot resistance introgressed from wild Helianthus petiolaris into cultivated sunflower (Helianthus annuus L.) using an advanced backcross population.

Sclerotinia sclerotiorum is a serious pathogen causing severe basal stalk rot (BSR) disease on cultivated sunflower (Helianthus annuus L.) that leads to significant yield losses due to insufficient resistance. The wild annual sunflower species H. petiolaris, commonly known as prairie sunflower is known for its resistance against this pathogen. Sunflower resistance to BSR is quantitative and determined by many genes with small effects on the resistance phenotype. The objective of this study was to identify loci governing BSR resistance derived from H. petiolaris using a quantitative trait loci (QTL) mapping approach. BSR resistance among lines of an advanced backcross population (AB-QTL) with 174 lines developed from a cross of inbred line HA 89 with H. petiolaris PI 435843 was determined in the field during 2017-2019, and in the greenhouse in 2019. AB-QTL lines and the HA 89 parent were genotyped using genotyping-by-sequencing and a genetic linkage map was developed spanning 997.51 cM and using 1,150 SNP markers mapped on 17 sunflower chromosomes. Highly significant differences (p<0.001) for BSR response among AB-QTL lines were observed disease incidence (DI) in all field seasons, as well as disease rating (DR) and area under the disease progress curve (AUDPC) in the greenhouse with a moderately high broad-sense heritability (H 2) of 0.61 for the tested resistance parameters. A total of 14 QTL associated with BSR resistance were identified on nine chromosomes, each explaining a proportion of the phenotypic variation ranging from 3.5% to 28.1%. Of the 14 QTL, eight were detected for BSR resistance in the field and six were detected under greenhouse conditions. Alleles conferring increased BSR resistance were contributed by the H. petiolaris parent at 11 of the 14 QTL.

Variability and selection of tomato Sletr1-2 mutants backcross population to improve yield and fruit quality

Enhancing fruit quality and yield is the goal of creating superior tomato varieties. One area of concern for breeders is fruit shelf life. To produce better offspring, hybridization attempts to combine the traits of two parents, but the results can be unpredictable. Backcrossing can create stable offspring with desired traits and eliminate undesirable characteristics from the parent plants. This study aimed to assess the diversity among advanced backcross strains and to obtain backcross genotypes carrying mutant alleles that have a high level of similarity with the superior parental lines. The study was carried out from April to September 2019 at the Indonesian Vegetable Research Institute’s screen house. In this experiment, two populations of backcrossing comprised of commercial parents of ‘Intan’ and ‘Mutiara’ varieties as well as backcross genotypes including MBC3F1 32, MBC3F1 32.2, MBC3F1 32.3, MBC3F1 37.2, MBC3F1 45.3, MBC3F1 37.1, IBC3F1 34.2, IBC3F1 37.2, and IBC3F1 34 were utilized. A variance analysis was used to determine genotype diversity. The genetic similarity between parents and offspring was determined using cluster analysis, and a t-test was used to compare mean performance values. According to the results, the BC3F1 populations of ‘Intan’, and ‘Mutiara’ exhibited a high degree of phenotypic variability. Cluster analysis and a t-test revealed that the IBC3F1.37.2 and IBC3F1.45.3 genotypes resembled ‘Intan’, furthermore MBC3F1.34 and MBC3F1.34.2 had strikingly similar characteristics to ‘Mutiara’. Therefore, several individual plants carrying the Sletr1-2 mutant allele in these four genotypes can further assemble longer tomato fruit shelf life and high-yielding varieties.

Advanced backcross QTL analysis and comparative mapping with RIL QTL studies and GWAS provide an overview of QTL and marker haplotype diversity for resistance to Aphanomyces root rot in pea (Pisum sativum).

Aphanomyces euteiches is the most damaging soilborne pea pathogen in France. Breeding of pea resistant varieties combining a diversity of quantitative trait loci (QTL) is a promising strategy considering previous research achievements in dissecting polygenic resistance to A. euteiches. The objective of this study was to provide an overview of the diversity of QTL and marker haplotypes for resistance to A. euteiches, by integrating a novel QTL mapping study in advanced backcross (AB) populations with previous QTL analyses and genome-wide association study (GWAS) using common markers. QTL analysis was performed in two AB populations derived from the cross between the susceptible spring pea variety "Eden" and the two new sources of partial resistance "E11" and "LISA". The two AB populations were genotyped using 993 and 478 single nucleotide polymorphism (SNP) markers, respectively, and phenotyped for resistance to A. euteiches in controlled conditions and in infested fields at two locations. GWAS and QTL mapping previously reported in the pea-Aphanomyces collection and from four recombinant inbred line (RIL) populations, respectively, were updated using a total of 1,850 additional markers, including the markers used in the Eden x E11 and Eden x LISA populations analysis. A total of 29 resistance-associated SNPs and 171 resistance QTL were identified by GWAS and RIL or AB QTL analyses, respectively, which highlighted 10 consistent genetic regions confirming the previously reported QTL. No new consistent resistance QTL was detected from both Eden x E11 and Eden x LISA AB populations. However, a high diversity of resistance haplotypes was identified at 11 linkage disequilibrium (LD) blocks underlying consistent genetic regions, especially in 14 new sources of resistance from the pea-Aphanomyces collection. An accumulation of favorable haplotypes at these 11 blocks was confirmed in the most resistant pea lines of the collection. This study provides new SNP markers and rare haplotypes associated with the diversity of Aphanomyces root rot resistance QTL investigated, which will be useful for QTL pyramiding strategies to increase resistance levels in future pea varieties.

Ultra-High-Density Genetic Maps of Jatropha curcas × Jatropha integerrima and Anchoring Jatropha curcas Genome Assembly Scaffolds

Genetic maps facilitate an understanding of genome organization and the mapping of genes and QTLs for traits of interest. Our objective was to develop a high-density genetic map of Jatropha and anchoring scaffolds from genome assemblies. We developed two ultra-high-density genetic linkage maps of Jatropha curcas × Jatropha intergerrima using a backcross (BC1) population using SNP, AFLP and SSR markers. First, SNPs were identified through genotyping-by-sequencing (GBS). The polymorphic SNPs were mapped to 3267 Jat_r4.5 scaffolds and 484 Wu_JatCur_1.0 scaffolds, and then these genomic scaffolds were mapped/anchored to the genetic linkage groups along with the AFLP and SSR markers for each genome assembly separately. We successfully mapped 7284 polymorphic SNPs, and 54 AFLP and SSR markers on 11 linkage groups using the Jat_r4.5 genomic scaffolds, resulting in a genome length of 1088 cM and an average marker interval of 0.71 cM. We mapped 7698 polymorphic SNPs, and 99 AFLP and SSR markers on 11 linkage groups using the Wu_JatCur_1.0 genomic scaffolds, resulting in a genome length of 870 cM and an average marker interval of 1.67 cM. The mapped SNPs were annotated to various regions of the genome, including exon, intron and intergenic regions. We developed two ultra-high-density linkage maps anchoring a high number of genome scaffolds to linkage groups, which provide an important resource for the structural and functional genomics as well as for molecular breeding of Jatropha while also serving as a framework for assembling and ordering whole genome scaffolds.

Genetic variability and correlation studies of yield and yield related attributes in BC2F2 population of sweet corn

The present study was carried out to study the genetic variability, heritability, genetic advance as percent of mean, correlation, skewness, and kurtosis of the backcross population BC2F2 derived from the cross SCM-Se-Sh-2 × DBT 17. High heritability coupled with high genetic advance was observed for cob girth, number of kernels per row, cob weight, 100 grain weight and single plant yield, suggesting additive gene action and selection of progenies based on this trait will result in higher genetic gain. Association studies showed that single plant yield was positively correlated with all yield attributes, viz., cob weight, number of kernels per row, 100 grain weight and plant height. These results depicted that cob weight, numbers of kernels per row and 100 grain weight may be used as reliable criteria for improving grain yield. Single plant yield, 100 grain weight, cob weight and plant height showed positive skewness and platykurtic distribution. This indicates that traits are governed by a large number of genes, and most of the genes exhibited complementary gene action, suggesting rigorous selection to achieve genetic gain. Overall, it is seen that this population has potential for yield improvement with enriched beta-carotene content.

Genetic Dissection of Morphological Variation Between Cauliflower and a Rapid Cycling Brassica oleracea line.

To improve resolution to small genomic regions and sensitivity to small-effect loci in the identification of genetic factors conferring the enlarged inflorescence and other traits of cauliflower while also expediting further genetic dissection, 104 near-isogenic introgression lines (NIILs) covering 78.56% of the cauliflower genome, were selected from an advanced backcross population using cauliflower [B. oleracea var. botrytis L., mutant for Orange gene (ORG)] as the donor parent and a rapid cycling line (TO1434) as recurrent parent. Subsets of the advanced backcross population and NIILs were planted in the field for eight seasons, finding 141 marker-trait associations for 15 leaf-, stem- and flower-traits. Exemplifying the usefulness of these lines, we delineated the previously-known flower color gene to a 4.5 MB interval on C3; a gene for small plant size to a 3.4 MB region on C8; and a gene for large plant size and flowering time to a 6.1 MB region on C9. This approach unmasked closely linked QTL alleles with opposing effects (on chr. 8), and revealed both alleles with expected phenotypic effects and effects opposite the parental phenotypes. Selected B. oleracea NIILs with short generation time add new value to widely used research and teaching materials.

Introgression of Stay Green Quantitative Trait Locus (QTLS) into Elite Sorghum Variety by MABC

Sorghum is a major staple food crop for the people in semi-arid areas of Asia. Post-flowering drought is a global constraint of sorghum production. The study aimed to improve stay-green characteristics of GS-23 sorghum variety by transferring stg3A and stg3B QTL`s respectively from donor genotypes K260 and K359w of ICRISAT by using marker assisted backcrossing. The experimental material comprised of six basic generations P1, P2, F1, F2, BC1 and BC2 developed from crossing GS-23 × K260 and GS-23 × K359 were genotyped using a set of 133 SNPs and 79 SSR markers. Whereas, 53 polymorphic SNPs among parents and backcross F1s for stay green trait at maturity were used to track introgression of stay green trait. Similarly,10 SSR markers were found to be polymorphic were used to track introgression of stay green trait i.e., stay green trait QTL`s stg3A and stg3B from donor parent K260 and K359w respectively in GS-23 background across backcross population and to identify plants that were homozygous for the desired allele. In which 02 SNPs SnpSB0039 and SnpSB0093 were identified as polymorphic for both K260 and K359w. Whereas 17 SNPs identified polymorphic for K260 and 34 SNPs for K359w. These SNPs were validated in both F1 and BC1F1 populations of both the crosses. Similarly, out of 10 SSR markers utilized, 02 SSRs were identified polymorphic to K260 and 02 SSRs were identified for K359w. Whereas 06 SSRs were polymorphic to both the parents K260 and K359w. Prominent 02 SSRs viz., Xtxp 141_Fam and Xgap84_Vic found to be more reliable and polymorphic to both the parents. These SSRs were validated in both F1 and BC1F1 populations of both the crosses. The genotypic analysis revealed the presence of favorable alleles in homozygous conditions at markers loci associated with stg3A and stg3B QTL`s in BC populations, suggesting successful introgression of stay green QTLs from the donor parents to the recurrent parent. Therefore, our study demonstrated the utility of marker-assisted backcrossing for drought tolerance improvement of locally adapted sorghum variety.

QTL Mapping of Tiller Number in Korean Japonica Rice Varieties.

Tiller number is an important trait associated with yield in rice. Tiller number in Korean japonica rice was analyzed under greenhouse conditions in 160 recombinant inbred lines (RILs) derived from a cross between the temperate japonica varieties Odae and Unbong40 to identify quantitative trait loci (QTLs). A genetic map comprising 239 kompetitive allele-specific PCR (KASP) and 57 cleaved amplified polymorphic sequence markers was constructed. qTN3, a major QTL for tiller number, was identified at 132.4 cm on chromosome 3. This QTL was also detected under field conditions in a backcross population; thus, qTN3 was stable across generations and environments. qTN3 co-located with QTLs associated with panicle number per plant and culm diameter, indicating it had pleiotropic effects. The qTN3 regions of Odae and Unbong40 differed in a known functional variant (4 bp TGTG insertion/deletion) in the 5' UTR of OsTB1, a gene underlying variation in tiller number and culm strength. Investigation of variation in genotype and tiller number revealed that varieties with the insertion genotype had lower tiller numbers than those with the reference genotype. A high-resolution melting marker was developed to enable efficient marker-assisted selection. The QTL qTN3 will therefore be useful in breeding programs developing japonica varieties with optimal tiller numbers for increased yield.

Linkage drag analysis in three Aegilops speltoides introgressions carrying Sr47 in modern durum and hard red spring wheat germplasm.

Yield and quality tests of wheat lines derived from RWG35 show they carry little, or no linkage drag and are the preferred source of Sr47 for stem rust resistance. Three durum wheat (Triticum turgidum L. subsp. durum) lines, RWG35, RWG36, and RWG37 carrying slightly different Aegilops speltoides introgressions, but each carrying the Sr47 stem rust resistance gene, were backcrossed to three durum and three hard red spring (HRS) wheat (Triticum aestivum L.) cultivars to produce 18 backcross populations. Each population was backcrossed to the recurrent parent six times and prepared for yield trials to test for linkage drag. Lines carrying the introgression (S-lines) were compared to euploid sibling lines (W-lines) and their parent. Yield trials were conducted from 2018 to 2021 at three locations. Three agronomic and several quality traits were studied. In durum, lines derived from RWG35 had little or no linkage drag. Lines derived from RWG36 and RWG37 still retained linkage drag, most notably involving yield and thousand kernel weight, but also test weight, falling number, kernel hardness index, semolina extract, semolina protein content, semolina brightness, and peak height. In HRS wheat, the results were more complex, though the general result of RWG35 lines having little or no linkage drag and RWG36 and RWG37 lines retaining linkage drag still applied. But there was heterogeneity in the Glenn35S lines, and Linkert lines had problems combining with the Ae. speltoides introgressions. We concluded that introgressions derived from RWG35 either had eliminated linkage drag or any negative effects were minor in nature. We recommend that breeders who wish to incorporate Sr47 into their cultivars should work exclusively with germplasm derived from RWG35.

Identification of new resistance source for Sclerotinia stem rot in backcross population of B. juncea + S. alba allohexaploids: key to manage disease through host resistance

Identification of new resistance source for Sclerotinia stem rot in backcross population of B. juncea + S. alba allohexaploids: key to manage disease through host resistance

Variation in the Root System Architecture of Peach × (Peach × Almond) Backcrosses.

The spatial arrangement and growth pattern of root systems, defined by the root system architecture (RSA), influences plant productivity and adaptation to soil environments, playing an important role in sustainable horticulture. Florida's peach production area covers contrasting soil types, making it necessary to identify rootstocks that exhibit soil-type-specific advantageous root traits. In this sense, the wide genetic diversity of the Prunus genus allows the breeding of rootstock genotypes with contrasting root traits. The evaluation of root traits expressed in young seedlings and plantlets facilitates the early selection of desirable phenotypes in rootstock breeding. Plantlets from three peach × (peach × almond) backcross populations were vegetatively propagated and grown in rhizoboxes. These backcross populations were identified as BC1251, BC1256, and BC1260 and studied in a completely randomized design. Scanned images of the entire root systems of the plantlets were analyzed for total root length distribution by diameter classes, root dry weight by depth horizons, root morphological components, structural root parameters, and root spreading angles. The BC1260 progeny presented a shallower root system and lower root growth. Backcross BC1251 progeny exhibited a more vigorous and deeper root system at narrower root angles, potentially allowing it to explore and exploit water and nutrients in deep sandy entisols from the Florida central ridge.

Genetic dissection of N use efficiency using maize inbred lines and testcrosses

Although the use of heterosis in maize breeding has increased crop productivity, the genetic causes underlying heterosis for nitrogen (N) use efficiency (NUE) have been insufficiently investigated. In this study, five N-response traits and five low-N-tolerance traits were investigated using two inbred line populations (ILs) consisting of recombinant inbred lines (RIL) and advanced backcross (ABL) populations, derived from crossing Ye478 with Wu312. Both populations were crossed with P178 to construct two testcross populations. IL populations, their testcross populations, and the midparent heterosis (MPH) for NUE were investigated. Kernel weight, kernel number, and kernel number per row were sensitive to N level and ILs showed higher N response than did the testcross populations. Based on a high-density linkage map, 138 quantitative trait loci (QTL) were mapped, each explaining 5.6%–38.8% of genetic variation. There were 52, 34 and 52 QTL for IL populations, MPH, and testcross populations, respectively. The finding that 7.6% of QTL were common to the ILs and their testcross populations and that 11.7% were common to the MPH and testcross population indicated that heterosis for NUE traits was regulated by non-additive and non-dominant loci. A QTL on chromosome 5 explained 27% of genetic variation in all of the traits and Gln1-3 was identified as a candidate gene for this QTL. Genome-wide prediction of NUE traits in the testcross populations showed 14%–51% accuracy. Our results may be useful for clarifying the genetic basis of heterosis for NUE traits and the candidate gene may be used for genetic improvement of maize NUE.

Fine mapping and gene silencing pinpoint Capana10g002229 as a strong candidate gene regulating the deciduous character of ripe pepper fruit (Capsicum spp.).

The pepper S locus, which controls the deciduous character of ripe fruit, was first fine mapped into an interval with a physical length of ~ 38.03kb on chromosome P10. Capana10g002229, encoding a polygalacturonase, was proposed as a strong candidate gene based on sequence comparison, expression pattern analysis and virus-induced gene silencing (VIGS). The deciduous character of ripe fruit, which is controlled by the dominant S locus, is a domesticated trait with potential value in the pepper processing industry (Capsicum spp.). However, the gene associated with the S locus has not been identified. Here, one major QTL designated S10.1 was detected by using the F2 population (n = 155) derived from BA3 (Capsicum annuum) × YNXML (Capsicum frutescens) and was further verified in an intraspecific backcross population (n = 254) derived from the cross between BB3 (C. annuum) and its wild relative Chiltepin (C. annuum var. glabriusculum) with BB3 as the recurrent parent. Then, a large BC1F2 population derived from the self-pollination of BB3 × (BB3 × Chiltepin) individuals and comprising 4217 individuals was used to screen the recombinants, and the S locus was ultimately delimited into a 38.03-kb region on chromosome P10 harbouring four annotated genes. Capana10g002229, encoding a polygalacturonase (PG), was proposed as the best candidate gene for S based on sequence comparison and expression pattern analyses. Downregulation of Capana10g002229 in fruits through VIGS significantly delayed fruit softening and abscission from the fruit-receptacle junction. Taken together, the results show that Capana10g002229 could be regarded as a strong candidate gene associated with the S locus in pepper. These findings not only lay a foundation for deciphering the molecular mechanisms underlying pepper domestication but also provide a strategy for genetic improvement of the deciduous character of ripe fruit using a marker-assisted selection approach.

Fine mapping of QTL conferring resistance to calcareous soil in mungbean reveals VrYSL3 as candidate gene for the resistance

Iron is a crucial nutrient for biological functions in plants. High-pH and calcareous soil is a major stress causing iron deficiency chlorosis (IDC) symptoms and yield losses in crops. Use of calcareous soil-tolerance genetic resources is the most effective preventative method to combat the effects of high-pH and calcareous soils. A previous study using a mungbean recombinant inbred line (RIL) population of the cross Kamphaeg Saen 2 (KPS2; IDC susceptible) × NM-10–12 identified a major quantitative trait locus (QTL), qIDC3.1, which controls resistance and explains more than 40% of IDC variation. In this study, we fine-mapped qIDC3.1 and identified an underlying candidate gene. A genome wide association analysis (GWAS) using 162 mungbean accessions identified single nucleotide polymorphisms (SNPs) on chromosome 6; several SNPs were associated with soil plant analysis development (SPAD) values and IDC visual scores of mungbeans planted on calcareous soil, respectively. These SNPs corresponded to qIDC3.1. Using the same RIL population as in the previous study and an advanced backcross population developed from KPS2 and IDC-resistant inbred line RIL82, qIDC3.1 was further confirmed and fine-mapped to an interval of 217 kilobases harboring five predicted genes, including LOC106764181 (VrYSL3), which encodes a yellow stripe1-like-3 (YSL3) protein, YSL3 is involved in iron deficiency resistance. Gene expression analysis revealed that VrYSL3 was highly expressed in mungbean roots. In calcareous soil, expression of VrYSL3 was significantly up-regulated, and it was more obviously upregulated in the roots of RIL82, than in those of KPS2. Sequence comparison of VrYSL3 between the RIL82 and KPS2 revealed four SNPs that result in amino acid changes in the VrYSL3 protein and a 20-bp insertion/deletion in the promoter where a cis-regulatory element resides. Transgenic Arabidopsis thaliana plants overexpressing VrYSL3 showed enhanced iron and zinc contents in the leaves. Taken together, these results indicate that VrYSL3 is a strong candidate gene responsible for calcareous soil resistance in mungbean.

Combined QTL mapping on bi-parental immortalized heterozygous populations to detect the genetic architecture on heterosis.

From bi-parental pure-inbred lines (PIL), immortalized backcross (i.e., IB1 and IB2, representing the two directions of backcrossing) and F2 (i.e., IF2) populations can be developed. These populations are suitable for genetic studies on heterosis, due to the present of both homozygous and heterozygous genotypes, and in the meantime allow repeated phenotyping trials across multiple locations and years. In this study, we developed a combined approach of quantitative trait locus (QTL) mapping, when some or all of the four immortalized populations (i.e., PIL, IB1, IB2, and IF2) are available. To estimate the additive and dominant effects simultaneously and accurately, suitable transformations are made on phenotypic values from different populations. When IB1 and IB2 are present, summation and subtraction are used. When IF2 and PIL are available, mid-parental values and mid-parental heterosis are used. One-dimensional genomic scanning is performed to detect the additive and dominant QTLs, based on the algorithm of inclusive composite interval mapping (ICIM). The proposed approach was applied to one IF2 population together with PIL in maize, and identified ten QTLs on ear length, showing varied degrees of dominance. Simulation studies indicated the proposed approach is similar to or better than individual population mapping by QTL detection power, false discovery rate (FDR), and estimated QTL position and effects.