Introgression Of Alleles Research Articles

Registration of ten soft red winter waxy wheat germplasm lines

AbstractWaxy wheat carries null alleles at the three Wx loci encoding the granule‐bound starch synthase I and produces endosperm starch composed of mostly amylopectin with less than 5% amylose. The USDA‐ARS developed and released 10 soft red winter (SRW) waxy wheat germplasm lines, SWQL‐K‐ABD‐1, SWQL‐K‐ABD‐2, SWQL‐K‐ABD‐3, SWQL‐K‐ABD‐4, SWQL‐K‐ABD‐5, SWQL‐W‐ABD‐1, SWQL‐W‐ABD‐2, SWQL‐W‐ABD‐3, SWQL‐W‐ABD‐4, SWQL‐W‐ABD‐5 (Reg. no. GP‐1061 to Reg. no. GP‐1070, PI 698615 to PI 698624), by the introgression of the null alleles at the Wx loci from a soft spring waxy wheat germplasm to two SRW wheat cultivars adapted to the eastern region of the United States. Ten waxy wheat germplasm lines were selected from BC4 or BC5 derivatives of the SRW wheat cultivars ‘Kristy’ and ‘Wilson’ (99ID490/5*Kristy and 99ID490/6*Wilson). The kernel hardness and starch amylose content of 10 SRW waxy wheat germplasm lines ranged from 34.1 to 63.1 and 2.5–7.2%, respectively, while those of wild types and SRW wheat parents carrying the functional alleles at the three Wx loci ranged from 33.9 to 60.4 and 26.8–27.0%, respectively. The starch of the waxy wheat germplasm lines exhibited a lower pasting temperature, higher peak viscosity, and lower setback value than the corresponding wild‐type wheat starch. These waxy wheat germplasm lines would be useful for the development of SRW wheat cultivars carrying different null alleles at the Wx loci that are adapted to the eastern region of the United States.

A signature of Neanderthal introgression on molecular mechanisms of environmental responses.

Ancient human migrations led to the settlement of population groups in varied environmental contexts worldwide. The extent to which adaptation to local environments has shaped human genetic diversity is a longstanding question in human evolution. Recent studies have suggested that introgression of archaic alleles in the genome of modern humans may have contributed to adaptation to environmental pressures such as pathogen exposure. Functional genomic studies have demonstrated that variation in gene expression across individuals and in response to environmental perturbations is a main mechanism underlying complex trait variation. We considered gene expression response to in vitro treatments as a molecular phenotype to identify genes and regulatory variants that may have played an important role in adaptations to local environments. We investigated if Neanderthal introgression in the human genome may contribute to the transcriptional response to environmental perturbations. To this end we used eQTLs for genes differentially expressed in a panel of 52 cellular environments, resulting from 5 cell types and 26 treatments, including hormones, vitamins, drugs, and environmental contaminants. We found that SNPs with introgressed Neanderthal alleles (N-SNPs) disrupt binding of transcription factors important for environmental responses, including ionizing radiation and hypoxia, and for glucose metabolism. We identified an enrichment for N-SNPs among eQTLs for genes differentially expressed in response to 8 treatments, including glucocorticoids, caffeine, and vitamin D. Using Massively Parallel Reporter Assays (MPRA) data, we validated the regulatory function of 21 introgressed Neanderthal variants in the human genome, corresponding to 8 eQTLs regulating 15 genes that respond to environmental perturbations. These findings expand the set of environments where archaic introgression may have contributed to adaptations to local environments in modern humans and provide experimental validation for the regulatory function of introgressed variants.

Whole-genome resequencing reveals genetic structure and introgression in Pudong White pigs

Pudong White (PDW) pigs, historically originating from Shanghai, are the only Chinese indigenous pigs characterised by their completely white coats, with the exception of Rongchang pigs. However, there is limited information concerning their overall genetic structure or relationship with other breeds, especially the East Chinese (ECN) and European pigs. To uncover the genetic structure, selection signatures, and potential exotic introgression in PDW pigs, we sampled 15 PDW pigs using whole-genome sequencing (~20×). We then conducted in-depth population genetic analyses in 320 pigs from 27 global pig groups, namely, European wild boars, Chinese wild boars, and outgroup. Neighbour-joining tree and principal component analysis confirmed that PDW pigs belonged to the ecotype of ECN pigs. Both f3, D-statistics, and structure analysis showed that PDW pigs shared apparent alleles with Large White (LW) pigs. Three statistics, rIBD, a haplotype heat map and copy number variation, further indicated that PDW pigs shared apparent alleles with LW pigs at the KIT Proto-Oncogene, Receptor Tyrosine Kinase (KIT) and PARG-MARCHF8 loci, suggesting that the lineage of European pigs in PDW originated from LW pigs. After further detecting the KIT mutations in different pig breeds, PDW was confirmed to have the same duplication region 1, duplication region 2, and the splicing mutation on intron 17 of KIT as LW pigs that determine the white coat colour phenotype in European white pigs. We hypothesised that LW pigs were imported to China ∼110–160 years ago according to the admixture time estimate and then crossed with ECN pigs, resulting in the introgression of the KIT alleles that produce the white coat colour phenotype in the PDW pig breed. To our knowledge, this study presents the first thorough description of the genetic structure of PDW pigs via whole-genome resequencing data; moreover, the results provide a basis for the national project for the conservation of this unique Chinese local population.

Impact of selection regime and introgression on deltamethrin resistance in the arbovirus vector Aedes aegypti - a comparative study between contrasted situations in New Caledonia and French Guiana.

Pyrethroid insecticides such as deltamethrin have been massively used against Aedes aegypti leading to the spread of resistance alleles worldwide. In an insecticide resistance management context, we evaluated the temporal dynamics of deltamethrin resistance using two distinct populations carrying resistant alleles at different frequencies. Three different scenarios were followed: a continuous selection, a full release of selection, or a repeated introgression with susceptible individuals. The responses of each population to these selection regimes were measured across five generations by bioassays and by monitoring the frequency of knockdown resistance (kdr) mutations and the transcription levels and copy number variations of key detoxification enzymes. Knockdown resistance mutations, overexpression and copy number variations of detoxification enzymes as a mechanism of metabolic resistance to deltamethrin was found and maintained under selection across generations. On comparison, the release of insecticide pressure for five generations did not affect resistance levels and resistance marker frequencies. However, introgressing susceptible alleles drastically reduced deltamethrin resistance in only three generations. The present study confirmed that strategies consisting to stop deltamethrin spraying are likely to fail when the frequencies of resistant alleles are too high and the fitness cost associated to resistance is low. In dead-end situations like in French Guiana where alternative insecticides are not available, alternative control strategies may provide a high benefit for vector control, particularly if they favor the introgression of susceptible alleles in natural populations. © 2021 Society of Chemical Industry.

Drought and crop yield.

Episodes of water shortage occur in most agricultural regions of the world. Their durations and intensities increase, and their seasonal timing alters with changing climate. During the ontogenic cycle of crop plants, each development stage, such as seed germination, seedling establishment, vegetative root and shoot growth, flowering, pollination and seed and fruit development, is specifically sensitive to dehydration. Desiccation threatens yield and leads to specific patterns, depending on the type of crop plant and the harvested plant parts, e.g. leafy vegetables, tubers, tap roots or fruits. This review summarizes the effects of drought stress on crop plants and relates the dehydration-dependent yield penalty to the harvested organ and tissue. The control of shoot transpiration and the reorganization of root architecture are of core importance for maintaining proper plant water relationships. Upon dehydration, the provision and partitioning of assimilates and the uptake and distribution of nutrients define remaining growth activity. Domestication of crops by selection for high yield under high input has restricted the genetic repertoire for achieving drought stress tolerance. Introgression of suitable alleles from wild relatives into commercial cultivars might improve the ability to grow with less water. Future research activities should focus more on field studies in order to generate more realistic improvements to crops. Robotic field phenotyping should be integrated into genetic mapping for the identification of relevant traits.

Single nucleotide replacement in the Atlantic salmon genome using CRISPR/Cas9 and asymmetrical oligonucleotide donors

BackgroundNew breeding technologies (NBT) using CRISPR/Cas9-induced homology directed repair (HDR) has the potential to expedite genetic improvement in aquaculture. The long generation time in Atlantic salmon makes breeding an unattractive solution to obtain homozygous mutants and improving the rates of perfect HDR in founder (F0) fish is thus required. Genome editing can represent small DNA changes down to single nucleotide replacements (SNR). This enables edits such as premature stop codons or single amino acid changes and may be used to obtain fish with traits favorable to aquaculture, e.g. disease resistance. A method for SNR has not yet been demonstrated in salmon.ResultsUsing CRISPR/Cas9 and asymmetrical ODNs, we were able to perform precise SNR and introduce a premature stop codon in dnd in F0 salmon. Deep sequencing demonstrated up to 59.2% efficiency in single embryos. In addition, using the same asymmetrical ODN design, we inserted a FLAG element into slc45a2 and dnd, showing high individual perfect HDR efficiencies (up to 36.7 and 32.7%, respectively).ConclusionsIn this work, we demonstrate that precise SNR and knock-in (KI) can be performed in F0 salmon embryos using asymmetrical oligonucleotide (ODN) donors. We suggest that HDR-induced SNR can be applied as a powerful NBT, allowing efficient introgression of favorable alleles and bypassing challenges associated with traditional selective breeding.

Screening of CRISPR/Cas9 gRNA for mimicking Powdery Mildew resistant MLO ol-2 mutant.

Powdery Mildew (PM) caused by fungal pathogen Oidium neolycopersici (O. neolycopersici) affects both greenhouse and field-grown tomato production. Resistance to PM disease can be achieved by selective inactivation of Mildew Resistance Locus O (MLO) genes encoding heptahelical transmembrane domains, which confer susceptibility to fungal pathogens. Natural loss-of-function mutation is a 19 base pair (bp) deletion in the SlMLO1 gene locus responsible for fungal resistance in S. lycopersicum var. cerasiforme. Introgression of these resistance alleles through breeding into elite varieties is possible. However, this is a long and labour-intensive process and has limitations due to linkage drag. Nonetheless, recent developments in the field of genome editing technology particularly CRISPR/Cas9 systems allows quick, effective and accurate genome modification at the target gene locus. Therefore, it is of interest to determine the efficacy and exact deletion that mimics the natural ol-2 (Slmlo1) mutation present in wild tomatoes using CRISPR/Cas9. 947 putative guide RNAs (gRNAs) were designed using Cas9 variants to broaden Protospacer Adjacent Motif (PAM) compatibility and to enhance DNA specificity against the SlMLO1 locus. 60 out of 947 gRNAs were selected based on the recognition of the PAM sequence, the MIT specificity ranking, the off-target sites, their distance from the 19bp natural ol-2 mutation, the secondary structure of the gRNAs, and their minimum free energy. In depth analysis of these 60 gRNAs helped in the selection of the top five gRNAs based on the above-mentioned criteria. These gRNAs are useful for introducing deletions identical to natural ol-2 mutants and impart resistance against fungal pathogen O. neolycopersici in cultivated tomato crops.

Improving grain appearance of erect-panicle japonica rice cultivars by introgression of the null gs9 allele

The panicle architecture and grain size of rice affect not only grain yield but also grain quality, especially grain appearance. The erect-panicle (EP) trait controlled by the qpe9-1/dep1 allele has been widely used in high-yielding japonica rice breeding, but usually accompanied with moderate appearance of milled rice. The null gs9 allele shows a good potential for improving grain shape and appearance. However, GS9 and qPE9-1/DEP1 loci are tightly linked, and their interaction is unclear, which obviously restricts their utilization in modern rice breeding. In the present study, comparative analyses of protein and mRNA levels revealed that GS9 and qPE9-1 function independently. Three near-isogenic lines (NILs) carrying various allelic combinations of these two loci, NIL (gs9/qpe9-1), NIL (GS9/qPE9-1) and NIL (gs9/qPE9-1), in the EP japonica cultivar 2661 (GS9/qpe9-1) background were developed for genetic interaction analysis. GS9 and qPE9-1 had additive effects on determining grain size, and the null gs9 allele could decrease grain chalkiness and improve grain appearance without affecting plant and panicle architecture in EP japonica cultivars. Additionally, introgression lines (ILs) developed in another released EP japonica cultivar Wuyujing 27 (WYJ27) background showed the same additive effect and the feasibility of utilizing the gs9 allele to improve grain appearance quality in high-yielding EP cultivars. This study provides an effective strategy for rice breeders to improve rice grain appearance in EP japonica and related cultivars.

Introgression of ASIP and TYRP1 Alleles Explains Coat Color Variation in Valais Goats.

The Valais Blackneck goat is a Swiss goat breed with a characteristic coat color phenotype. Before the revision of the breed standard in 1938, 4 different color varieties of Valais goats were known. Besides Blackneck animals resembling the modern breed standard, the brown and white Copperneck goat, the white Capra Sempione, and the greyish Grüenochte comprised the historic Valais goats. The brown pigmentation of Copperneck goats had previously been traced back to an introgression of a mutant TYRP1 allele from Toggenburg goats. In the present study, we identified additional introgression events of distinct ASIP alleles causing the remaining 2 rare coat color patterns within the Valais Blackneck goat breed. We identified the introgression of the AWt allele from Appenzell or Saanen goats in white Capra Sempione goats. Similarly, introgression of the Apc allele from Peacock goats resulted in the greyish Grüenochte phenotype. These results demonstrate past hybridization events between breeds that are separated today. A perfect genotype-phenotype association in 393 Valais goats supported the causality of the genotyped variants for the different coat color phenotypes. Our study gives insights into the introgression of functionally relevant copy number variant (CNV) alleles controlling pigmentation between goat breeds with strikingly different coat color patterns.

<strong>Population structure and association studies for reproductive stage salinity tolerance in rice (<em>Oryza sativa</em> L.)</strong>

Salinity is a major abiotic stress responsible for yield loss in rice as it severely affects various yield contributing traits. Rice is categorised as salt sensitive crop and it is important to identify genomic regions associated to salinity tolerance. In the present study, association mapping was performed to investigate the functional relationship between microsatellite markers and salinity related traits in a set of 180 diverse rice accessions. Association analysis was carried out by employing mixed linear model (MLM) approach. Population structure analysis revealed four subgroups in entire study panel while the admixture level ranged from 0.7-57.2%. A total of 22 marker-trait associations were discovered and four marker-trait associations explained phenotypic variation (R2) greater than 10%. Furthermore, 7 markers were found close to the candidate genes loci. Several markers were significantly associated with more than one trait, suggesting pleiotropic effects. The phenotypic variation explained by associated markers ranged from 2.92 to 18.50%. Comparative genomic search revealed that associated markers were close to candidate genes which play significant role in signal transduction, metabolic pathways and transcription factor activity. The significant associations identified in the present study could be used to improve salt tolerance in rice with introgression of favourable alleles through marker assisted breeding.

Genic introgression from an invasive exotic fungal forest pathogen increases the establishment potential of a sibling native pathogen

Significant hybridization between the invasive North American fungal plant pathogen Heterobasidion irregulare and its Eurasian sister species H. annosum is ongoing in Italy. Whole genomes of nine natural hybrids were sequenced, assembled and compared with those of three genotypes each of the two parental species. Genetic relationships among hybrids and their level of admixture were determined. A multi-approach pipeline was used to assign introgressed genomic blocks to each of the two species. Alleles that introgressed from H. irregulare to H. annosum were associated with pathways putatively related to saprobic processes, while alleles that introgressed from the native to the invasive species were mainly linked to gene regulation. There was no overlap of allele categories introgressed in the two directions. Phenotypic experiments documented a fitness increase in H. annosum genotypes characterized by introgression of alleles from the invasive species, supporting the hypothesis that hybridization results in putatively adaptive introgression. Conversely, introgression from the native into the exotic species appeared to be driven by selection on genes favoring genome stability. Since the introgression of specific alleles from the exotic H. irregulare into the native H. annosum increased the invasiveness of the latter species, we propose that two invasions may be co-occurring: the first one by genotypes of the exotic species, and the second one by alleles belonging to the exotic species. Given that H. irregulare represents a threat to European forests, monitoring programs need to track not only exotic genotypes in native forest stands, but also exotic alleles introgressed in native genotypes.

Four Species Linked by Three Hybrid Zones: Two Instances of Repeated Hybridization in One Species Group (Genus Liolaemus)

Hybridization is an evolutionary process that can generate diverse outcomes, such as reinforcing species boundaries, generating new species, or facilitating the introgression of locally-adapted alleles into new genomic backgrounds.Liolaemusis a highly diverse clade of South American lizards with ~260 species and as many as ten new species are described each year. PreviousLiolaemusstudies have detected gene flow and introgression among species using phylogenetic network methods and/or through comparisons of nuclear and mitochondrial DNA patterns, yet no study has systematically studied hybrid zones betweenLiolaemusspecies. Here, we compared three hybrid zones between four species in theLiolaemus fitzingeriigroup of lizards in Central Argentina where two species,L. melanopsandL. xanthoviridis, each hybridize with two other species (L. shehuenandL. fitzingerii). We sampled three transects that were each ~120 km in length and sequenced both mitochondrial and genome-wide SNP data for 267 individuals. In our analyses of nuclear DNA, we also compared bi-allelic SNPs to phased alleles (50 bp RAD loci). Population structure analyses confirmed that boundaries separating species are sharp, and all clines are <65 km wide. Cline center estimates were consistent between SNPs and phased alleles, but cline width estimates were significantly different with the SNPs producing wider estimates. The mitochondrial clines are narrower and shifted 4–20 km southward in comparison to the nuclear clines in all three hybrid zones, indicating that either each of the species has sex-biased dispersal (males northward or females southward), the population densities are unequal, or that the hybrid zones are moving north over time. These comparisons indicate that some patterns of hybridization are similar across hybrid zones (mtDNA clines all narrower and shifted to the south), whereas cline width is variable. Hybridization in theL. fitzingeriigroup is common and geographically localized; further studies are needed to investigate whether hybrid zones act as hard species boundaries or promoters of speciation through processes such as reinforcement. Nonetheless, this study provides insights into both biotic and abiotic mechanisms helping to maintain species boundaries within the specioseLiolaemussystem.

Genome-Wide Association Study Identified Novel Candidate Loci/Genes Affecting Lodging Resistance in Rice.

Lodging reduces rice yield, but increasing lodging resistance (LR) usually limits yield potential. Stem strength and leaf type are major traits related to LR and yield, respectively. Hence, understanding the genetic basis of stem strength and leaf type is of help to reduce lodging and increase yield in LR breeding. Here, we carried out an association analysis to identify quantitative trait locus (QTLs) affecting stem strength-related traits (internode length/IL, stem wall thickness/SWT, stem outer diameter/SOD, and stem inner diameter/SID) and leaf type-associated traits (Flag leaf length/FLL, Flag leaf angle/FLA, Flag leaf width/FLW, leaf-rolling/LFR and SPAD/Soil, and plant analyzer development) using a diverse panel of 550 accessions and evaluated over two years. Genome-wide association study (GWAS) using 4,076,837 high-quality single-nucleotide polymorphisms (SNPs) identified 89 QTLs for the nine traits. Next, through “gene-based association analysis, haplotype analysis, and functional annotation”, the scope was narrowed down step by step. Finally, we identified 21 candidate genes in 9 important QTLs that included four reported genes (TUT1, OsCCC1, CFL1, and ACL-D), and seventeen novel candidate genes. Introgression of alleles, which are beneficial for both stem strength and leaf type, or pyramiding stem strength alleles and leaf type alleles, can be employed for LR breeding. All in all, the experimental data and the identified candidate genes in this study provide a useful reference for the genetic improvement of rice LR.

Unique venom proteins from Solenopsis invicta x Solenopsis richteri hybrid fire ants.

The Solenopsis venom protein 2 transcript was amplified, sequenced, probed, and analyzed from Solenopsis invicta x Solenopsis richteri hybrid ant colonies (hybrids) collected from across Tennessee to determine the extent of introgression of each parent allele (Solenopsis invicta venom protein 2 [Soli2] and Solenopsis richteri venom protein 2 [Solr2]). Chemotaxonomic analyses of venom alkaloids and cuticular hydrocarbons were used to categorize hybrid colonies and their relative relatedness to each parent species. Hybrid colonies were chosen randomly from each chemotaxonomic hybridization category, including “very near S. richteri,” “near S. richteri,” “near S. invicta,” and “very near S. invicta.” Lateral flow immunoassays for detection of the Soli2 and Solr2 venom proteins were largely in agreement with the chemotaxonomic analyses for the very near S. richteri (100% Solr2) and very near S. invicta (80% Soli2, 20% Soli2 + Solr2 detected in the sample) groups, while Soli2 and Solr2 were reported in 60% and 40% in the near S. invicta and near S. richteri chemotaxonomic groups. Analysis of transcripts from the hybrid colonies revealed a sequence with 100% identity to Soli2 (GenBank Accession L09560) and three unique sequences, which we identify as Solenopsis hybrid venom protein 2 (Solh2; GenBank Accession MT150127), Solenopsis hybrid truncated venom protein 2 (Solh2Tr97; Genbank Accession MT150129), and Solenopsis richteri venom protein 2, D to A change at position 69 (Solr2A69; GenBank Accession MT150128). The predicted open reading frame for Solh2 and Solh2Tr97 revealed sequences unique to hybrid ants, with Solh2Tr97an alternatively spliced form. A third unique sequence, Solr2A69, is likely the correct sequence for Solr2, which appears to have been published previously with a sequencing error (GenBank Accession P35776).

Adaptive introgression of the beta-globin cluster in two Andean waterfowl.

Introgression of beneficial alleles has emerged as an important avenue for genetic adaptation in both plant and animal populations. In vertebrates, adaptation to hypoxic high-altitude environments involves the coordination of multiple molecular and cellular mechanisms, including selection on the hypoxia-inducible factor (HIF) pathway and the blood-O2 transport protein hemoglobin (Hb). In two Andean duck species, a striking DNA sequence similarity reflecting identity by descent is present across the ~20 kb β-globin cluster including both embryonic (HBE) and adult (HBB) paralogs, though it was yet untested whether this is due to independent parallel evolution or adaptive introgression. In this study, we find that identical amino acid substitutions in the β-globin cluster that increase Hb-O2 affinity have likely resulted from historical interbreeding between high-altitude populations of two different distantly-related species. We examined the direction of introgression and discovered that the species with a deeper mtDNA divergence that colonized high altitude earlier in history (Anas flavirostris) transferred adaptive genetic variation to the species with a shallower divergence (A. georgica) that likely colonized high altitude more recently possibly following a range shift into a novel environment. As a consequence, the species that received these β-globin variants through hybridization might have adapted to hypoxic conditions in the high-altitude environment more quickly through acquiring beneficial alleles from the standing, hybrid-origin variation, leading to faster evolution.

Evaluation and Selection of Interspecific Lines of Groundnut (Arachis hypogaea L.) for Resistance to Leaf Spot Disease and for Yield Improvement.

Early and late leaf spot are two devastating diseases of peanut (Arachis hypogaea L.) worldwide. The development of a fertile, cross-compatible synthetic amphidiploid, TxAG-6 ([A. batizocoi × (A. cardenasii × A. diogoi)]4x), opened novel opportunities for the introgression of wild alleles for disease and pest resistance into commercial cultivars. Twenty-seven interspecific lines selected from prior evaluation of an advanced backcross population were evaluated for resistance to early and late leaf spot, and for yield in two locations in Ghana in 2006 and 2007. Several interspecific lines had early leaf spot scores significantly lower than the susceptible parent, indicating that resistance to leaf spot had been successfully introgressed and retained after three cycles of backcrossing. Time to appearance of early leaf spot symptoms was less in the introgression lines than in susceptible check cultivars, but the opposite was true for late leaf spot. Selected lines from families 43-08, 43-09, 50-04, and 60-02 had significantly reduced leaf spot scores, while lines from families 43-09, 44-10, and 63-06 had high pod yields. One line combined both resistance to leaf spot and high pod yield, and several other useful lines were also identified. Results suggest that it is possible to break linkage drag for low yield that accompanies resistance. However, results also suggest that resistance was diluted in many of the breeding lines, likely a result of the multigenic nature of resistance. Future QTL analysis may be useful to identify alleles for resistance and allow recombination and pyramiding of resistance alleles while reducing linkage drag.

An advanced lentil backcross population developed from a cross between Lens culinaris × L. ervoides for future disease resistance and genomic studies

AbstractGenetically accessible variation to some of the abiotic and biotic stresses are limited in the cultivated lentil (Lens culinaris Medik.) germplasm. Introgression of novel alleles from its wild relative species will be useful for enhancing the genetic improvement of the crop. L. ervoides, one of the wild relatives of lentil, is a proven source of disease resistance for the crop. Here we introduce a lentil advanced backcross (LABC-01) population developed in cultivar ‘CDC Redberry’ background, based on L. ervoides alleles derived from an interspecific recombinant inbred population, LR-59-81. Two-hundred and seventeen individuals of the LABC-01 population at BC2F3:4 generation were screened for the race 0 of anthracnose (Colletotrichum lentis) and stemphylium blight (Stemphylium botryosum) under controlled conditions. The population showed significant variations for both diseases and the transfer of resistance alleles into the elite cultivar was evident. It also segregated for other traits such as days to flowering, seed coat colour, seed coat pattern and flower colour. Overall, we showed that LABC-01 population can be used in breeding programmes worldwide to improve disease resistance and will be available as a valuable genetic resource for future genetic analysis of desired loci introgressed from L. ervoides.

Marker-assisted stacking of null Kunitz trypsin inhibitor and off-flavour generating lipoxygenase-2 in soybean

AbstractSoybean varieties genetically free from Kunitz trypsin inhibitor (KTI) and lipoxygenase-2 (Lox2) are desirable to increase human consumption, as the former is an antinutritional factor that affects protein digestibility while the latter is a principal contributor to off-flavour. In the present investigation, soybean genotypes free from both these undesirable components were developed by introgression of null allele of Lox2 from NRC109 (lox2lox2) into two KTI-free soybean genotypes derived from genotypes JS97-52 and NRC7. Foreground selection of plants in F1, F2, BC1F1, BC1F2, BC2F1 and BC2F2 generations developed from two cross combinations i.e. NRC7-derived KTI-free genotype (N7KTIF)×NRC109 (parental combination 1) and JS97-52 derived KTI free genotype (JKTIF)×NRC109 (parental combination 2) was performed using null allele specific markers and tightly linked simple sequence repeat (SSR) markers for both KTI and Lox2 genes for the identification of homozygous recessive (titilox2lox2) plants. Background selection was performed using 239 and 241 polymorphic SSR markers across the genome. This resulted in the development of 9 and 13 soybean lines stacked for null alleles of both KTI and Lox2 (titilox2lox2) exhibiting recurrent parent genome content more than 97 and 96%, respectively. Days-to-flowering, days-to-maturity, 100-seed weight and yield per plant of the stacked lines developed from both the parental combinations were at par with the respective recurrent parents.

Introgression, admixture, and selection facilitate genetic adaptation to high-altitude environments in cattle

Domestication and subsequent selection of cattle to form breeds and biological types that can adapt to different environments partitioned ancestral genetic diversity into distinct modern lineages. Genome-wide selection particularly for adaptation to extreme environments left detectable signatures genome-wide. We used high-density genotype data for 42 cattle breeds and identified the influence of Bos grunniens and Bos javanicus on the formation of Chinese indicine breeds that led to their divergence from India-origin zebu. We also found evidence for introgression, admixture, and migration in most of the Chinese breeds. Selection signature analyses between high-altitude (≥1800 m) and low-altitude adapted breeds (<1500 m) revealed candidate genes (ACSS2, ALDOC, EPAS1, EGLN1, NUCB2) and pathways that are putatively involved in hypoxia adaptation. Immunohistochemical, real-time PCR and CRISPR/cas9 ACSS2-knockout analyses suggest that the up-regulation of ACSS2 expression in the liver promotes the metabolic adaptation of cells to hypoxia via the hypoxia-inducible factor pathway. High altitude adaptation involved the introgression of alleles from high-altitude adapted yaks into Chinese Bos taurus taurus prior to their formation into recognized breeds and followed by selection. In addition to selection, adaptation to high altitude environments has been facilitated by admixture and introgression with locally adapted cattle populations.

Development of tetra-primer amplicon refractory mutation system (T-ARMS) strategy for identification of the dark green mutant allele in tomato.

The high pigment (hp) tomato mutants increase the fruit pigment content in tomato. The dark green (dg) tomato mutant belonging to this category contains the hp-2 dg recessive allele, resulting from a single nucleotide polymorphism (SNP) in the DEETIOLATED1 (DET1) gene. Naturally, identification of this functional SNP is crucial for introgression of this important mutant allele in suitable tomato cultivar(s). Hence, we developed a functional co-dominant marker for detecting the hp-2 dg mutant allele using the tetra-primer amplicon refractory mutation system (T-ARMS) strategy. Although the other available techniques for identification of SNPs require specialized costly equipments and reagents, our method allows the reliable identification of this functional SNP in time-, labour- and cost-effective manner, using standard molecular biology equipments and reagents. To the best of our knowledge, this is the first report of T-ARMS strategy for the detection of the dg mutant allele in tomato, which will definitely help in identification and introgression of this allele by the breeders, working even under limited fund and resources.