Candidate QTL Research Articles

Identification of candidate genes and genomic prediction for early heifer pregnancy in Nelore beef cattle

This study aimed to assess the influence of differential weighting in genomic regions harboring candidate causal loci on the prediction accuracy and inflation for early heifer pregnancy (heifers that calved up to 30 months of age) in Nellore (Bos indicus) heifers using the single step genomic BLUP model (ssGBLUP). Phenotypic records of 102,294 Nellore heifers born between 2010 and 2017 were used in this study. The pedigree dataset harbored information from 176,107 animals born between 1998 and 2017, including 5,145 sires and 35,705 dams. Seven different models for genomic prediction were defined by combining the SNP weights obtained in the iterations (1st and 2nd) of the weighted single step GWAS (ssw1GBLUP and ssw2GBLUP) or candidate QTLs reported in the literature. Hence, the lambda (λ) values estimated in the WssGWAS were used to weight the SNPs adjacent to the candidate regions or QTL previously reported in the literature. To estimate the genetic parameters and perform the WssGWAS and WssGBLUP for early heifer pregnancy, a single-trait Bayesian analysis considering a threshold animal model was used. Accuracy, bias, and inflation parameters were evaluated in the validation subset based on the linear regression (LR) method. Genomic windows of ten consecutive SNPs that explained more than 0.5% of the additive genetic variance were selected to explore and determine possible candidate genes. Among the identified genes, we can highlight the PGRMC2, TENM3, GRIP1, TMEM45A, and KLF3, given their roles in endocrine fertility, expression of contractile proteins, average daily gain, dry matter intake, and fat deposition. Several genomic regions associated with QTL related to early heifer pregnancy were identified. The identification of such regions and the respective candidate genes associated with sexual precocity and fertility would contribute to improve the genetic knowledge regarding early sexual precocity of Nellore cattle. The prediction accuracy increased roughly 25.6% using the ssGBLUP compared to BLUP models. The prediction accuracy with the WssGBLUP when incorporating weighted SNPs with the λ values obtained in the 1st (ssw1GBLUP) and 2nd iteration (ssw2GBLUP) of the WssGWAS was higher (∼18%) than that described for the ssGBLUP model. The inflation also increased the weighting of the most relevant SNPs obtained with the GWAS, most likely overestimating the GEBV. The models that weighted SNPs close to QTLs reported in the literature yielded to less biased and deflated predictions compared to ssw1GBLUP and ssw2GBLUP models. Genomic selection is a feasible alternative for genomic evaluation of early heifer pregnancy in Nellore beef cattle by increasing the prediction accuracy of young animals. In addition, the use of information obtained from the WssGWAS is an alternative to increase reliability and reduce genomic prediction bias. Therefore, the results obtained herein indicate that it is possible to improve the prediction accuracy and reduce the bias of genomic prediction by using genomic information and differentially weighted genomic regions harboring candidate QTLs previously reported in the literature for early heifer pregnancy.

A QTL on chromosome 17 identified by Genome-Wide Association Mapping controls postharvest cold tolerance of Cucurbita pepo L.

The worldwide cultivated Cucurbita pepo L. is one of the most diverse species in the plant kingdom. In this study, chilling tolerance over a wide range of cultivars was characterized to discover the allelic variants to improving the postharvest quality of the immature fruit during cold storage. For this purpose, fruits from 126 accessions of worldwide origin have been evaluated for weight loss and chilling injury after 3, 7 and 14 days of cold storage, classifying them into tolerant, partially tolerant, and sensitive accessions. To verify this classification, antioxidant capacity and lipid peroxidation (MDA) of contrasting accessions (tolerant vs. sensitive) were assessed. The antioxidant capacity significantly decreased during cold storage in the sensitive accessions, while it was maintained in tolerant accessions. Additionally, the sensitive accessions presented a higher accumulation of MDA during this period. Finally, a GWAS analysis using GBS data available in CuGenDBv2, combined with weight loss percentage data, led to the identification of a candidate QTL located on chromosome 17 that regulates postharvest cold tolerance in zucchini. The region contains four SNPs whose alternative alleles were significantly associated with lower weight loss percentage and chilling injury indices during cold storage. Two SNPs are located in the 3' UTR region of the gene CpERS1, a gene involved in ethylene perception. The other two SNPs generate missense mutations in the coding region of a Pectin methyl esterase inhibitor gene (CpPMI). The role of this QTL and these variants in chilling tolerance is discussed.

GWAS as a tool for detecting SNPs in cattle to study their relationship to reproduction, productivity, growth, behavior, diseases

The fundamental goal of animal husbandry is the cost-effective production of human food from healthy animals, which includes production, reproduction. The method of whole-genome association study (GWAS) is actively used in various fields, including agriculture. Genome-wide association analyzes were generated as an identifier for genomic variations associated with economically significant traits in different livestock species. This method of genomic selection provides new priorities for improving the productive and reproductive qualities of livestock.The purpose of this review article is a comprehensive analysis of the current state of GWAS in cattle, focusing on the identification of SNPs associated with reproduction, productivity, growth, behavior and genetically determined diseases. The scope of the article covers the study of GWAS results worldwide, both in dairy and beef cattle, with special emphasis on the identification of candidate genes, QTL and genome regions related to the direction of productivity. Additionally, the organization of this review will include a classification of GWAS results based on the specific traits studied, providing a comprehensive overview of the genetic determinants of reproduction, growth, behavior, and disease traits in cattle.

Weighted genomic prediction for growth and carcass-related traits in Nelore cattle.

This study aimed to assess the impact of differential weighting in genomic regions harboring candidate causal loci on the genomic prediction accuracy and dispersion for growth and carcass-related traits in Nelore cattle. The dataset contained 168 793 phenotypic records for adjusted weight at 450 days of age (W450), 83 624 for rib eye area (REA), 24 480 for marbling (MAR) and 82 981 for subcutaneous backfat thickness (BFT) and rump fat thickness (RFT). The pedigree harbored information from 244 254 animals born between 1977 and 2016, including 6283 sires and 50 742 dams. Animals (n=7769) were genotyped with the low-density panel (Clarifide® Nelore 3.0), and the genotypes were imputed to a panel containing 735 044 markers. A linear animal model was applied to estimate the genetic parameters and to perform the weighted single-step genome-wide association study (WssGWAS). A total of seven models for genomic prediction were evaluated combining the SNP weights obtained in the iterations of the WssGWAS and the candidate QTL. The heritability estimated for W450 (0.35) was moderate, and for carcass-related traits, the estimates were moderate for REA (0.27), MAR (0.28) and RFT (0.28), and low for BFT (0.18). The prediction accuracy for W450 incorporating reported QTL previously described in the literature along with different SNPs weights was like those described for the default ssGBLUP model. The use of the ssGWAS to weight the SNP effects displayed limited advantages for the REA prediction accuracy. Comparing the ssGBLUP with the BLUP model, a meaningful improvement in the prediction accuracy from 0.09 to 0.63 (700%) was observed for MAR. The highest prediction accuracy was obtained for BFT and RFT in all evaluated models. The application of information obtained from the WssGWAS is an alternative to reduce the genomic prediction dispersion for growth and carcass-related traits, except for MAR. Furthermore, the results obtained herein pointed out that is possible to improve the prediction accuracy and reduce the genomic prediction dispersion for growth and carcass-related traits in young animals.

Whole-genome SNP markers reveal runs of homozygosity in indigenous cattle breeds of Pakistan

The runs of homozygosity (ROH) were identified in 14 Pakistani cattle breeds (n = 105) by genotyping with the Illumina 50 K SNP BeadChip. These breeds were categorized into Dairy, Dual, and Draft breeds based on their utility and production performance. We identified a total of 10,936 ROHs which mainly consisted of a high number of shorter segments (1–4 Mb). Dairy group exhibited the highest level of inbreeding (FROH: 0.078 ± 0.028) while the lowest (FROH: 0.002 ± 0.008) was observed in Dual group. In 48 genomic regions identified with a high frequency of ROH, 207 genes were detected in the three breed groups. A substantially higher number of ROH islands detected in dairy breeds indicated the impact of the positive selection pressure over the years. Important candidate genes and QTL were detected in the ROH islands associated with economic traits like milk production, reproduction, meat, carcass, and health traits in dairy cattle.

Mapping of dwarfing QTL of Ari1327, a semi-dwarf mutant of upland cotton

BackgroundUpland Cotton (Gossypium hirsutum L.) has few cotton varieties suitable for mechanical harvesting. The plant height of the cultivar is one of the key features that need to modify. Hence, this study was planned to locate the QTL for plant height in a 60Co γ treated upland cotton semi-dwarf mutant Ari1327.ResultsInterestingly, bulk segregant analysis (BSA) and genotyping by sequencing (GBS) methods exhibited that candidate QTL was co-located in the region of 5.80–9.66 Mb at D01 chromosome in two F2 populations. Using three InDel markers to genotype a population of 1241 individuals confirmed that the offspring’s phenotype is consistent with the genotype. Comparative analysis of RNA-seq between the mutant and wild variety exhibited that Gh_D01G0592 was identified as the source of dwarfness from 200 genes. In addition, it was also revealed that the appropriate use of partial separation markers in QTL mapping can escalate linkage information.ConclusionsOverwhelmingly, the results will provide the basis to reveal the function of candidate genes and the utilization of excellent dwarf genetic resources in the future.

Multi-Environmental Genetic Analysis of Grain Size Traits Based on Chromosome Segment Substitution Line in Rice (Oryza sativa L.)

Grain size traits are critical agronomic traits which directly determine grain yield, but the genetic bases of these traits are still not well understood. In this study, a total of 154 chromosome segment substitution lines (CSSLs) population derived from a cross between a japonica variety Koshihikari and an indica variety Nona Bokra was used to investigate grain length (GL), grain width (GW), length-width ratio (LWR), grain perimeter (GP), grain area (GA), and thousand grain weight (TGW) under four environments. QTL mapping analysis of six grain size traits was performed by QTL IciMapping 4.2 with an inclusive composite interval mapping (ICIM) model. A total of 64 QTLs were identified for these traits, which mapped to chromosomes 1, 2, 3, 4, 6, 7, 8, 10, 11, and 12 and accounted for 1.6%–27.1% of the total phenotypic variations. Among these QTLs, thirty-six loci were novel and seven QTLs were identified under four environments. One locus containing the known grain size gene, qGL3/GL3.1/OsPPKL1, also have been found. Moreover, five pairs of digenic epistatic interactions were identified except for GL and GP. These findings will facilitate fine mapping of the candidate gene and QTL pyramiding to genetically improve grain yield in rice.

Association analysis in a germplasm panel of watermelon (Citrullus lanatus (Thunb.) Matsum and Nakai) predominantly from India confirms a QTL conferring resistance to Fusarium oxysporum f. sp. niveum race 1

ABSTRACT An experiment was conducted to screen a germplasm panel comprising 331 Citrullus sp. accessions against Fon race 1 for identification of resistance and to validate a candidate QTL (Fon-1) region conferring resistance. The accessions were screened through artificial inoculation of spore suspension during the rainy season of 2017. A genotype with ≥67% survival at 28 days post-inoculation (DPI) was categorised as resistant. A total of 41 accessions were identified to possess resistance to Fon race 1. Of these, nine accessions were highly resistant (mean survival ≥90% at 28 DPI). The reactions of these accessions were also confirmed in another bioassay. These accessions need deployment in future watermelon breeding programmes. Six markers, located within the previously reported Fon-1 locus, were tested for their association with resistance and among them, InDel-4, located in the 5ʹ promoter region of an ABC Transporter protein, was found to be most significantly associated and explained maximum phenotypic variation, proving its relevance for marker-assisted introgression of resistance to Fon race 1 into elite commercial backgrounds of watermelon.

Single-step genome-wide association study uncovers known and novel candidate genomic regions for endocrine and classical fertility traits in Swedish Red and Holstein dairy cows

• In this study, the novel endocrine and classical fertility traits of Swedish dairy herds were evaluated using in-line milk progesterone profiles in ssGWAS approach. • The genomic regions associated with the endocrine and classical traits were identified. • A total of 20 QTLs, of which 18 of them are novel, were detected. The novel QTL regions embedded candidate genes ( ATG7, GPI, GJA1, PPP3CA, ECT2, ARHGAP20, PHLDB1, CACNA1D, CCNE1, CDH13, PLD1, FBN2, KIF3A, FGF12, KCNMB2, MAN1A1, KCNN2, SMAD6, MAPK8IP1, PHF21A, LPXN, MMRN1, KCNIP4, NID2, PCDHGA8, GRIA1, PCDHGB4, PHLDB2, STXBP5L, PTPRR, SRGAP1, SNX27, SPTA1, S100A10, TBC1D20 and ITCH ) that are associated with endocrine and classical fertility traits. • The candidate QTL regions provide insights into the genetic basis of endocrine and classical fertility in SR and Holstein dairy breeds. In a study aiming to identify candidate genomic regions associated with endocrine and classical fertility traits in Swedish Red (SR) and Holstein cows, data on 3955 lactations in 1164 SR and 1672 Holstein cows were examined. The dataset comprised milk progesterone (P4) levels ( n = 341,212) in 14 Swedish herds, automatically collected and analyzed in-line using the DeLaval Herd Navigator™. Endocrine traits studied were: days from calving to commencement of luteal activity (C-LA), first luteal phase length (LPL), length of first inter-luteal interval, length of first inter-ovulatory interval (IOI), luteal activity during the first 60 DIM, and proportion of samples with luteal activity during the first 60 DIM. Classical fertility traits based on insemination data were also investigated, such as days from calving to last insemination and calving interval. A total of 180 SR and 312 Holstein cows were genotyped with a low-density SNP chip and imputed to 50 K. Single-step genome-wide association (ssGWAS) was used to explore candidate genomic regions associated with fertility traits. A mixed linear single-trait animal model was fitted, considering season and parity as fixed effects and animal and permanent environment as random effects. The results revealed 990 and 415 SNPs above the threshold (-log ( p -value) ≥4) for SR and Holstein cows, respectively. The breeds shared only eight SNPs significantly associated with fertility traits. Annotation analysis revealed 281 SNPs located in 241 genes. Functional enrichment analysis using DAVID tools reduced the number to 80 genes, which were mediated in various biological processes and KEGG pathways in multiple functions, including folliculogenesis, embryogenesis, uterine growth and development, immune response, and ovarian cysts. Of the 80 genes, 67 were associated with fertility traits in SR cows and 13 in Holstein. Most genes were associated with LPL and IOI in SR cows, but in Holstein the only association with an endocrine trait was with C-LA. Twenty QTL regions that embedded 40 genes were associated with fertility traits in both breeds. All the QTLs detected, except at BTA2 and BTA19 are novel QTL regions that were not reported previously. These novel QTL regions embedded the candidate genes that include ARHGAP20, PHLDB1, CACNA1D, ATG7, CCNE1, GPI, CDH13, ECT2, PLD1, FBN2, KIF3A, FGF12, KCNMB2, GJA1, MAN1A1, KCNN2, SMAD6, MAPK8IP1, PHF21A, LPXN, MMRN1, KCNIP4, NID2, PCDHGA8, GRIA1, PCDHGB4, PHLDB2, STXBP5L, PPP3CA, PTPRR, SRGAP1, SNX27, SPTA1, S100A10, TBC1D20 and ITCH. The candidate regions may help to improve genetic progress in female fertility if used in selection decisions. A challenge for future research is to determine why different regions seem relevant for different traits and breeds, and the practical implications for genomic selection.

Mapping resistance to WBNV through CIM, JICIM and QTL-meta analysis approaches in watermelon

Since past three decades, tospoviruses have been impacting severe economic losses causing heavy yield reduction in many crops. Watermelon bud necrosis disease caused by watermelon bud necrosis orthotospovirus (WBNV) is one such tospovirus that has emerged as a devastating disease of watermelon in India. Efforts are underway to develop resistant varieties at ICAR-IIHR and in the current study, efforts have been made to integrate WBNV resistance QTLs from wide range of populations and screening experiments for a comprehensive analysis through QTL-meta-analysis and joint inclusive composite interval mapping (JICIM). Comprehensive analyses for QTLs suggested MQTL_WBNV_2.1 on Chromosome 2, MQTL_WBNV_3.2 on Chromosome 3 and MQTL_WBNV_7.2 on Chromosome 7 as potential candidate QTL regions for WBNV resistance. The genes identified in these QTLs viz., Cla020173 encoding PR-1, Cla001733 encoding pathogenesis-related transcriptional factor and ethylene-responsive transcription (ERF), Cla001590 and Cla001591 genes encoding acid phosphatase, Cla019476 encoding TIR-NBS and Cla011771 belonging to TIR-NBS-LRR gene family may be investigated as putative candidates for WBNV resistance in watermelon. The results obtained in present study may provide way for functional validation of putative candidates and development of suitable markers for marker assisted selection (MAS) of WBNV resistance in watermelon.

SNP markers associated with resistance to frosty pod and black pod rot diseases in an F1 population of Theobroma cacao L.

Economically, cacao (Theobroma cacao L.) is a major tropical commodity for the Americas; however, severe losses due to Moniliophthora roreri (Cif. and Par.), which causes frosty pod rot (FPR), and Phytophthora spp., which causes black pod rot (BPR), have reduced cacao production in the Americas. The objectives of this study are to (i) re-confirm the QTL using different marker set; (ii) discover new QTL associated with FPR and BPR resistance using SNP markers; and (iii) find genes in the candidate QTL regions. At CATIE in Turrialba, Costa Rica, an F1 mapping population of cacao was obtained by crossing “POUND 7,” a clone moderately susceptible to FPR and resistant to BPR, with “UF 273,” resistant to FPR and highly susceptible to BPR. A total of 179 F1 progeny were fingerprinted with 5149 SNP markers and a dense linkage map composed of 10 linkage groups was developed using 2910 polymorphic SNP markers. Also segregating F1 trees were screened for resistance to FPR and BPR diseases. Seven QTL previously reported on chromosomes 2, 7, and 8 for FPR resistance and on chromosomes 4, 8, and 10 for BPR resistance were confirmed. Additionally, eight QTL were identified for FPR resistance (chromosomes 4, 9, and 10) and BPR resistance (chromosome 2). The expression of genes commonly associated with plant defense and disease resistance that are located within the identified QTL was confirmed.

QTL identification for salt tolerance related traits at the seedling stage in indica rice using a multi-parent advanced generation intercross (MAGIC) population

Salinity is one of the most important abiotic stresses, which seriously affects rice production. In this study, a multiparent advanced generation intercross population, DC1, derived from intercrossing four elite indica varieties, was used to identify QTLs conferring salt tolerance. The whole population and four parents were genotyped with a 55K rice SNP array. A total of 7 QTLs delineated from 186 significant marker-trait associations were detected on chromosomes 1, 2, 5 and 9, which accounted for 7.42–9.38% of the total phenotypic variations. Among these QTLs, one novel QTL (qRRL2) on chromosome 2 for relative root length and one multi-trait QTL (qSLST1/qRDSW1/qRB1) on chromosome 1 affecting shoot length, root dry weight and root biomass under salt treatment were detected. Gene expression analysis revealed that a transcription factor gene (LOC_Os01g66280) within the multi-trait QTL is the potential candidate gene for salt tolerance. Interestingly, we identified one significant nonsynonymous SNP in the coding region of this candidate gene. These results will facilitate fine mapping of the candidate gene and QTL pyramiding to genetically improve salt tolerance in rice.

Functional annotation and Bayesian fine-mapping reveals candidate genes for important agronomic traits in Holstein bulls

A hundred years of data collection in dairy cattle can facilitate powerful studies of complex traits. Cattle GWAS have identified many associated genomic regions. With increasing numbers of cattle sequenced, fine-mapping of causal variants is becoming possible. Here we imputed selected sequence variants to 27,214 Holstein bulls that have highly reliable phenotypes for 35 production, reproduction, and body conformation traits. We performed single-marker scans for the 35 traits and multi-trait tests of the three trait groups, revealing 282 candidate QTL for fine-mapping. We developed a Bayesian Fine-MAPping approach (BFMAP) to integrate fine-mapping with functional enrichment analysis. Our fine-mapping identified 69 promising candidate genes, including ABCC9, VPS13B, MGST1, SCD, MKL1, CSN1S1 for production, CHEK2, GC, KALRN for reproduction, and TMTC2, ARRDC3, ZNF613, CCND2, FGF6 for conformation traits. Collectively, these results demonstrated the utility of BFMAP, identified candidate genes, and enhanced our understanding of the genetic basis of cattle complex traits.

Disentangling Genetic Variation for Resistance and Endurance to Scuticociliatosis in Turbot Using Pedigree and Genomic Information.

Selective breeding for improving host responses to infectious pathogens is a promising option for disease control. In fact, disease resilience, the ability of a host to survive or cope with infectious challenge, has become a highly desirable breeding goal. However, resilience is a complex trait composed of two different host defence mechanisms, namely resistance (the ability of a host to avoid becoming infected or diseased) and endurance (the ability of an infected host to survive the infection). While both could be targeted for genetic improvement, it is currently unknown how they contribute to survival, as reliable estimates of genetic parameters for both traits obtained simultaneously are scarce. A difficulty lies in obtaining endurance phenotypes for genetic analyses. In this study, we present the results from an innovative challenge test carried out in turbot whose design allowed disentangling the genetic basis of resistance and endurance to Philasterides dicentrarchi, a parasite causing scuticociliatosis that leads to substantial economic losses in the aquaculture industry. A noticeable characteristic of the parasite is that it causes visual signs that can be used for disentangling resistance and endurance. Our results showed the existence of genetic variation for both traits (heritability = 0.26 and 0.12 for resistance and endurance, respectively) and for the composite trait resilience (heritability = 0.15). The genetic correlation between resistance and resilience was very high (0.90) indicating that both are at a large extent the same trait, but no significant genetic correlation was found between resistance and endurance. A total of 18,125 SNPs obtained from 2b-RAD sequencing enabled genome-wide association analyses for detecting QTLs controlling the three traits. A candidate QTL region on linkage group 19 that explains 33% of the additive genetic variance was identified for resilience. The region contains relevant genes related to immune response and defence mechanisms. Although no significant associations were found for resistance, the pattern of association was the same as for resilience. For endurance, one significant association was found on linkage group 2. The accuracy of genomic breeding values was also explored for resilience, showing that it increased by 12% when compared with the accuracy of pedigree-based breeding values. To our knowledge, this is the first study in turbot disentangling the genetic basis of resistance and endurance to scuticociliatosis.

Identification of Early Tomato Fruit Ripening Loci by QTL-seq

QTL-seq has been successfully studied in identifying major QTLs, markers, and candidate genes associated with traits that are important for crop improvement. Tomato earliness is an economically important trait and is a major current research focus recently. This paper reports the identification of tomato early ripening fruit locus facilitated by QTL-seq using a novel next-generation sequencing technology. Two DNA pools of phenotypes of F2 offspring from crosses between the Bone ММ (early ripening fruit, P1) and 071-440 (late ripening fruit, P2) cultivars of (Solanum lycopersicum) were bulked for sequencing and alignment analysis. Sequencing results revealed 434 SNP markers on chromosome 11, a candidate QTL at position 52,048,208 bp (named er-fruit) and a candidate gene, Solyc11g071510.1.1. The “er-fruit” as confirmed by the traditional QTL method was related to the early fruit ripening trait in tomato. Additionally, BLAST analysis to known homologies for Solyc11g071510.1.1 gene encodes glycoside hydrolases (GHs). GHs are functionally associated with cell wall degradation, fruit softening and ripening. Thus, GHs may be important in fruit softening, stimulating early fruit ripening in tomato. Our results confirmed that QTL-seq is effective method to identify candidate QTL loci, candidate genes and candidate markers.

QTL mapping of male sterility and transmission pattern in progeny of Satsuma mandarin.

Seedlessness is one of the important traits in citrus breeding. Male sterility derived from Satsuma mandarin (Citrus unshiu) has been used in Japanese citrus breeding programs to obtain seedless cultivars. The efficiency of seedless cultivar breeding would be improved by developing a selection marker linked to seedlessness. In this study, we performed QTL mapping in ‘Okitsu No. 46’ × ‘Okitsu No. 56’ (O46-O56) crosses for the number of pollen grains per anther (NPG) and apparent pollen fertility (APF), two traits used as an index of male sterility, and detected a candidate QTL for NPG (MS-P1) on linkage group 8 with a significant LOD score (7.31) and 47% of variance explained. The QTL for APF (MS-F1) was detected on linkage group 6 with a significant LOD score (5.71) and 63.6% of variance explained. The role of both MS-P1 in reducing NPG and MS-F1 in decreasing APF were confirmed with the ‘Okitsu No.46’ × ‘Kara’ (O46-K) cross. Pedigree analysis inferred that both MS-P1 and MS-F1 in ‘Okitsu No. 46’ were derived from kunenbo (Citrus nobilis) through hassaku (C. hassaku) and ‘Sweet Spring’. Cytoplasm analysis revealed that both male-sterile ‘Sweet Spring’ and ‘Okitsu No. 46’ have cytoplasm derived from Kishu (C. kinokuni hort. ex Tanaka), but the cytoplasm of male-sterile kunenbo and hassaku were derived from other varieties rather than Kishu. These results suggest that MS-P1 and MS-F1 primarily reduce the NPG and decrease APF, but their expression requires a cytoplasm derived from Kishu. These findings will improve our understanding of the molecular mechanism of male sterility in citrus and help to develop a DNA marker for seedless breeding in citrus.

An ultra-high density map allowed for mapping QTL and candidate genes controlling dry latex yield in rubber tree

Rubber tree (Hevea brasiliensis) is the most important commercial producer of high-quality natural rubber in the world. Here, we identified 571,267 SNPs and 134,184 indels, and constructed the first ultra-high density genetic linkage map in rubber tree population. This map consisted of 6940 markers, with average marker density and recombination rate 0.30 cM and 0.97 cM/Mb, respectively. Then the whole genome QTL scanning for dry latex yield (DLY) trait were performed and seventeen repQTLs were obtained, among which qDFY-10 and qDFY-18-4 could explain up to 38.3% and 33.3% phenotypic variability, respectively. Numerous highly-promising QTL candidate genes were identified and then verified significant associated with DLY trait, including some first found in rubber tree, such as thioredoxin h, plastin-like protein, calmodulin binding protein, cytochrome-c oxidase and methylglutaconyl-CoA hydratase. This ultra-high density linkage map supplies a base for important traits QTL mapping and will be useful in the improvement of the assembly of genome sequences in rubber tree. The verification of candidate QTL genes showed great importance for understanding the possible function of mapped QTLs and the comprehensive relations between phenotypes and genotypes. Accurate QTL mapping will be enhance genetic studies that can be applied in rubber tree breeding programs.

Interval mapping for red/green skin color in Asian pears using a modified QTL-seq method

Pears with red skin are attractive to consumers and provide additional health benefits. Identification of the gene(s) responsible for skin coloration can benefit cultivar selection and breeding. The use of QTL-seq, a bulked segregant analysis method, can be problematic when heterozygous parents are involved. The present study modified the QTL-seq method by introducing a |Δ(SNP-index)| parameter to improve the accuracy of mapping the red skin trait in a group of highly heterozygous Asian pears. The analyses were based on mixed DNA pools composed of 28 red-skinned and 27 green-skinned pear lines derived from a cross between the ‘Mantianhong’ and ‘Hongxiangsu’ red-skinned cultivars. The ‘Dangshansuli’ cultivar genome was used as reference for sequence alignment. An average single-nucleotide polymorphism (SNP) index was calculated using a sliding window approach (200-kb windows, 20-kb increments). Nine scaffolds within the candidate QTL interval were in the fifth linkage group from 111.9 to 177.1 cM. There was a significant linkage between the insertions/deletions and simple sequence repeat markers designed from the candidate intervals and the red/green skin (R/G) locus, which was in a 582.5-kb candidate interval that contained 81 predicted protein-coding gene models and was composed of two subintervals at the bottom of the fifth chromosome. The ZFRI 130-16, In2130-12 and In2130-16 markers located near the R/G locus could potentially be used to identify the red skin trait in Asian pear populations. This study provides new insights into the genetics controlling the red skin phenotype in this fruit.

Rice Flowering Locus T 1 plays an important role in heading date influencing yield traits in rice

Important role of flowering genes in enhancing grain productivity in rice has become well recognized for a number of key genes regulating the florigen production, but little has been known for the two florigen genes themselves. In this study, pleiotropism of Rice Flowering Locus T 1 (RFT1), one of the two florigen genes in rice, was firstly evaluated using near isogenic lines (NILs) carrying RFT1 alleles from the indica rice cultivars Zhenshan 97 (ZS97) and Milyang 46, respectively, and then determined by transformation of the RFT1ZS97 allele into a japonica rice variety, Zhonghua 11. The RFT1ZS97 allele was shown to delay heading and increase plant height, grain weight, grain number and grain yield, indicating that RFT1 plays an important role in the growth and development of rice. This study has also validated the potential of using a new type of genetic resource, sequential residual heterozygotes (SeqRHs), for QTL fine-mapping. A step-by-step approach was employed for SeqRHs identification, NIL development and QTL fine-mapping. The heterozygous segments and candidate QTL regions were gradually narrowed down. Eventually, the QTL region was delimited to a 1.7 kb region containing a single gene.

Genome-wide association study for performance traits in chickens using genotype by sequencing approach

Performance traits are economically important and are targets for selection in breeding programs, especially in the poultry industry. To identify regions on the chicken genome associated with performance traits, different genomic approaches have been applied in the last years. The aim of this study was the application of CornellGBS approach (134,528 SNPs generated from a PstI restriction enzyme) on Genome-Wide Association Studies (GWAS) in an outbred F2 chicken population. We have validated 91.7% of these 134,528 SNPs after imputation of missed genotypes. Out of those, 20 SNPs were associated with feed conversion, one was associated with body weight at 35 days of age (P < 7.86E-07) and 93 were suggestively associated with a variety of performance traits (P < 1.57E-05). The majority of these SNPs (86.2%) overlapped with previously mapped QTL for the same performance traits and some of the SNPs also showed novel potential QTL regions. The results obtained in this study suggests future searches for candidate genes and QTL refinements as well as potential use of the SNPs described here in breeding programs.