Functional Genetic Diversity Analysis and Identification of Associated Simple Sequence Repeats and Amplified Fragment Length Polymorphism Markers to Drought Tolerance in Lentil (<Emphasis Type="Italic">Lens culinaris</Emphasis> ssp. <Emphasis Type="Italic">culinaris</Emphasis> Medicus) Landraces

Genetic diversity of 70 Mediterranean lentil (Lens culinaris ssp. culinaris Medicus) landraces was assessed using simple sequence repeats (SSRs) and amplified fragment length polymorphisms (AFLPs). These landraces were also assessed for variation in root and shoot traits and drought tolerance as estimated by relative water content (RWC), water losing rate (WLR) and wilting score (WS). Genetic diversity and clear differentiation of Moroccan landraces from those from northern Mediterranean regions (Italy, Turkey and Greece) were found. High genetic variation in root and shoot traits and traits related to drought tolerance was also observed. No relationship was found between drought tolerance of landraces and their geographic origin. Landraces with higher dry root biomass, chlorophyll content and root–shoot ratio were drought tolerant as evidenced by higher RWC and lower WLR and wilting severity. Kruskal–Wallis non-parametric test (K-W) was used to find SSRs and AFLPs associated with RWC, WLR and WS. Regression analysis showed six SSR and AFLP alleles explaining the highest phenotypic variation of RWC, WLR and WS (ranging from 21 to 50 % for SSRs and from 14 to 33 % for AFLPs). Functional genetic diversity analysis showed relationships between drought response of landraces and linked SSR and AFLP alleles to RWC, WLR and WS according to K-W test using canonical discriminant analysis. Our results confirm the feasibility of using association mapping to find DNA markers associated with drought tolerance in larger numbers of lentil landraces.     

Population structure analysis and association mapping of bacterial blight resistance in <Emphasis Type="Italic">indica</Emphasis> rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) accessions

Bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo), is a serious disease in rice production worldwide. To understand the genetic diversity of bacterial blight resistance a population consisting of 175 indica accessions from nine countries was collected and detected their association between SSR (Simple Sequence Repeat) markers and resistance to six bacterial races. The resistance phenotypes of various rice accessions were evaluated through artificial inoculation under controlled conditions in 2013 and 2014. Association analysis showed that 17 SSR markers were significantly associated with resistance to four bacterial races and the phenotypic variations explained (PVE) ranged from 7.43 to 15.05%. Among the 17 associated SSR markers, two SSR markers located in previously reported genes regions, and 15 SSR markers were newly identified in this study. These results validated a new approach to map resistance genes of rice to bacterial blight. These markers could be used for marker-assisted selection (MAS) in rice bacterial blight resistance breeding programs.     

Quantitative trait loci (QTL) mapping of blush skin and flowering time in a European pear (<Emphasis Type="Italic">Pyrus communis</Emphasis>) progeny of ‘Flamingo’ × ‘Abate Fetel’

Blush skin and flowering time are agronomic traits of interest to the Agricultural Research Council (ARC) Infruitec-Nietvoorbij pear breeding programme. The genetic control of these traits was investigated in the pear progeny derived from ‘Flamingo’ (blush cultivar) × ‘Abate Fetel’ (slightly blush) made up of 121 seedlings. Blush skin was scored phenotypically over three seasons and flowering time was scored over two seasons. A total of 160 loci from 137 simple sequence repeat (SSR) markers were scored in the progeny and used to construct parental genetic linkage maps. Quantitative trait loci (QTL) analysis revealed two QTLs for blush skin, a major QTL on linkage group (LG) 5 in ‘Flamingo’, and a major QTL on LG9 in ‘Abate Fetel’. Two SSR markers, NB101a and SAmsCO865954, were closely linked with the major QTL on LG5 in ‘Flamingo’, with alleles 139 bp and 462 bp in coupling, respectively. These markers were present in approximately 90% of the seedlings scored as good blush (class 4) based on the average data set. These two markers were used to genotype other pear accessions to validate the QTL on LG5 with the view of marker-assisted selection. Two candidate genes, MYB86 and UDP-glucosyl transferase, were associated with the QTL on LG5 and MYB21 and MYB39 were associated with the QTL on LG9. QTL analysis for flowering time revealed a major QTL located on LG9 in both parents. Marker GD142 with allele 161 bp from ‘Flamingo’ was present in approximately 88% of the seedlings that flowered earlier than either parent, based on the average data set. The QTLs and linked markers will facilitate marker-assisted selection for the improvement of these complex traits.     

Genetic variation of a global germplasm collection of chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.) including Italian accessions at risk of genetic erosion

Chickpea (Cicer arietinum L.) is one of the most important legumes worldwide. We addressed this study to the genetic characterization of a germplasm collection from main chickpea growing countries. Several Italian traditional landraces at risk of genetic erosion were included in the analysis. Twenty-two simple sequence repeat (SSR) markers, widely used to explore genetic variation in plants, were selected and yielded 218 different alleles. Structure analysis and hierarchical clustering indicated that a model with three distinct subpopulations best fits the data. The composition of two subpopulations, named K1 and K2, broadly reflects the commercial classification of chickpea in the two types desi and kabuli, respectively. The third subpopulation (K3) is composed by both desi and kabuli genotypes. Italian accessions group both in K2 and K3. Interestingly, this study highlights genetic distance between desi genotypes cultivated in Asia and Ethiopia, which respectively represent the chickpea primary and the secondary centres of diversity. Moreover, European desi are closer to the Ethiopian gene pool. Overall, this study will be of importance for chickpea conservation genetics and breeding, which is limited by the poor characterization of germplasm collection.     

Genetic diversity changes in Indian lentils over the times

The genetic diversity of 96 genotypes of lentil comprising 34 cultivars, 46 advanced breeding lines, and 16 germplasm lines were studied using 260 SSR markers. These markers generated a total of 749 alleles. The alleles/locus ranged from 2 to 16 with an average value of 2.87. Polymorphic information content varied from 0.02 to 0.91 with a mean of 0.30. Major allelic frequency ranged from 0.14 to 0.99 with a mean of 0.77. Studied genotypes were clustered into two groups according to their breeding history. Advanced breeding lines derived from exotic lines were clustered in one group, while another group accommodated most of the cultivars and advanced breeding lines with common cultivars in parentage. The germplasm lines were sub-clustered within first group. Cumulatively, first three principal components contributed 21.2% to the total variability. Advanced breeding lines showed higher number of alleles/locus and gene diversity (He) than other sets of genetic materials. In present study, no significant differences were observed between cultivars developed in different decadal groups for both NA and He. Moreover, genetic diversity changes between small and large seeded lentil cultivars were also found non-significant in this study. These findings showed that the use of alien genes can help to diversify active gene pool for developing improved new cultivars in lentil.     

A novel set of 223 EST-SSR markers in <Emphasis Type="Italic">Casuarina</Emphasis> L. ex Adans.: polymorphisms,cross-species transferability,and utility for commercial clone genotyping

Simple sequence repeat (SSR) markers are very useful for genetic applications in plants, but SSR resource for the important tree genus Casuarina L. ex Adans. is still limited. In this study, we report a novel set of 223 SSR markers in Casuarina developed from expressed sequence tag (EST) resource of GenBank. The 223 EST-SSR markers were polymorphic among 10 unrelated individuals of C. equisetifolia L. Johnson, with the number of alleles per locus (N_a), observed heterozygosity (H_o), expected heterozygosity (H_e), and polymorphic information content (PIC) averaging at 5.5, 0.72, 0.86, and 0.63, respectively. The rates of cross-species transferability ranged from 96.9% (C. glauca Sieber ex Sprengel) through 97.8% (C. cunninghamiana Miquel) to 99.1% (C. junghuhniana Miquel). Fifty-five C. equisetifolia clones widely planted in China were successfully genotyped with a subset of 20 EST-SSRs. These newly developed markers will have a great potential for genetic and breeding applications in Casuarina species and related taxa.     

Association mapping to discover significant marker-trait associations for resistance against fusarium wilt variant 2 in pigeonpea [<Emphasis Type="Italic">Cajanus cajan</Emphasis> (L.) Millspaugh] using SSR markers

Pigeonpea production is severely constrained by wilt disease caused by Fusarium udum. In the current study, we discover the putative genomic regions that control resistance response to variant 2 of fusarium wilt using association mapping approach. The association panel comprised of 89 diverse pigeonpea genotypes including seven varieties, three landraces and 79 germplasm lines. The panel was screened rigorously for 3 consecutive years (2013–14, 2014–15 and 2015–2016) against variant 2 in a wilt-sick field. A total of 65 pigeonpea specific hypervariable SSR markers (HASSRs) were screened representing seven linkage groups and 29 scaffolds of the pigeonpea genome. A total of 181 alleles were detected, with average values of gene diversity and polymorphism information content (PIC) of 0.55 and 0.47, respectively. Further analysis using model based (STRUCTURE) and distance based (clustering) approaches separated the entire pigeonpea collection into two distinct subgroups (K = 2). The marker trait associations (MTAs) were established based on three-year wilt incidence data and SSR dataset using a unified mixed linear model. Consequently, six SSR markers were identified, which were significantly associated with wilt resistance and explained up to 6% phenotypic variance (PV) across the years. Among these SSRs, HASSR18 was found to be the most stable and significant, accounting for 5–6% PV across the years. To the best of our knowledge, this is the first report of identification of favourable alleles for resistance to variant 2 of Fusarium udum in pigeonpea using association mapping. The SSR markers identified here will greatly facilitate marker assisted resistance breeding against fusarium wilt in pigeonpea.     

Genome-wide association study of heading and flowering dates and construction of its prediction equation in Chinese common wheat

Heading date is one of the most important traits in wheat breeding as it affects adaptation and yield potential. A genome-wide association study (GWAS) using the 90 K iSelect SNP genotyping assay indicated that a total of 306 loci were significantly associated with heading and flowering dates in 13 environments in Chinese common wheat from the Yellow and Huai wheat region. Of these, 105 loci were significantly correlated with both heading and flowering dates and were found in clusters on chromosomes 2, 5, 6, and 7. Based on differences in distribution of the vernalization and photoperiod genes among chromosomes, arms, or block regions, 13 novel, environmentally stable genetic loci were associated with heading and flowering dates, including RAC875_c41145_189 on 1DS, RAC875_c50422_299 on 2BL, and RAC875_c48703_148 on 2DS, that accounted for more than 20% phenotypic variance explained (PVE) of the heading/flowering date in at least four environments. GWAS and t test of a combination of SNPs and vernalization and photoperiod alleles indicated that the Vrn-B1, Vrn-D1, and Ppd-D1 genes significantly affect heading and flowering dates in Chinese common wheat. Based on the association of heading and flowering dates with the vernalization and photoperiod alleles at seven loci and three significant SNPs, optimal linear regression equations were established, which show that of the seven loci, the Ppd-D1 gene plays the most important role in modulating heading and flowering dates in Chinese wheat, followed by Vrn-B1 and Vrn-D1. Additionally, three novel genetic loci (RAC875_c41145_189, Excalibur_c60164_137, and RAC875_c50422_299) also show important effect on heading and flowering dates. Therefore, Ppd-D1, Vrn-B1, Vrn-D1, and the novel genetic loci should be further investigated in terms of improving heading and flowering dates in Chinese wheat. Further quantitative analysis of an F₁₀ recombinant inbred lines population identified a major QTL that controls heading and flowering dates within the Ppd-D1 locus with PVEs of 28.4% and 34.0%, respectively; this QTL was also significantly associated with spike length, peduncle length, fertile spikelets number, cold resistance, and tiller number.     

Genetic structure of the Russian populations of <Emphasis Type="Italic">Pyrenophora tritici-repentis</Emphasis>, determined by using microsatellite markers

The population genetic structure of plant pathogenic fungus Pyrenophora tritici-repentis was examined using microsatellite (SSR) markers. According to the geographical origin of the pathogen populations, they were designated as North Caucasian (S, 33 isolates), northwest (Nw, 39), and Omsk (Om, 43). The populations were analyzed at the nine most polymorphic SSR loci, at which 75 alleles were identified. To characterize the genetic variation within and between populations, the AMOVA algorithm as implemented in the Arlequin v. 3.5 software program was used. The number of alleles per locus ranged from 5 to 12 and their sizes varied within the range from 180 to 400 bp. The mean gene diversity at SSR loci was high for all populations (H = 0.58–0.75). The populations were considerably different in the frequencies of individual alleles of the SSR loci. Most isolates in the populations were represented by unique haplotypes. The within-population variation of the isolates at molecular markers was 86.4%; among the populations, 13.6%. Substantial interpopulation differences were found between the Om and S (F_st = 0.16) and between the Om and Nw (F_st = 0.20) populations, whereas between the S and Nw populations, these differences were small (F_st = 0.05). Thus, it was demonstrated that the population of P. tritici-repentis from Omsk oblast had the independent status of the geographical population; northwest and North Caucasian populations differed in the allelic diversity of SSR loci, and despite the low F_st value (0.05), they also belonged to independent geographical populations.     

Genetic analysis and localization of loci controlling leaf rust resistance of <Emphasis Type="Italic">Triticum aestivum</Emphasis> × <Emphasis Type="Italic">Triticum timopheevii</Emphasis> introgression lines

Introgressive lines resulting from crossing common wheat Triticum aestivum with the tetraploid T. timopheevii are characterized by effective resistance to leaf rust caused by Puccinia triticina Eriks. Molecular analysis using 350 specific simple sequence repeat (SSR) markers determined localization of the T. timopheevii genome in chromosomes 1A, 2A, 2B, 5A, 5B, and 6B. A population of F₂ offspring of crossing hybrid line 842-2 with common wheat cultivar Skala was obtained for mapping the loci controlling leaf rust resistance. Analysis of association of phenotypic and genotypic data by means of simple interval mapping (SIM) and composite interval mapping (CIM) has shown that the resistance of adult plants is determined by two loci in chromosomes 5B and 2A. The major locus QLr.icg-5B, transferred from T. timopheevii chromosome 5G mapped to the interval of microsatellite loci Xgwm408-Xgwm1257 controls 72% of the phenotypic variance of the trait. The other, minor locus QLr.icg-2A located to chromosome 2A at a distance of 10 cM from Xgwm312 accounts for 7% of the trait expression. Microsatellite markers located near these loci may be used for controlling the transfer of agronomically valuable loci when new lines and cultivars are created.     

A major QTL (<Emphasis Type="Italic">qFT12.1</Emphasis>) allele from wild soybean delays flowering time

Soybean is highly sensitive to photoperiod. To improve the adaptability and productivity of soybean, it is essential to understand the molecular mechanisms regulating flowering time. To identify new flowering time QTLs, we evaluated a BC₃F₅ population consisting of 120 chromosome segment substitution lines (CSSLs) over 2 years under field conditions. CSSLs were derived from a cross between the cultivated soybean cultivar Jackson and the wild soybean accession JWS156-1, followed by continuous backcrossing using Jackson as the recurrent parent. Four QTLs (qFT07.1, qFT12.1, qFT12.2, and qFT19.1) were detected on three chromosomes. Of these, qFT12.1 showed the highest effect, accounting for 36.37–38.27% of the total phenotypic variation over 2 years. This QTL was further confirmed in the F₇ recombinant inbred line population (n?=?94) derived from the same cross (Jackson × JWS156-1). Analysis of the qFT12.1 BC₃F₅ residual heterozygous line RHL509 validated the allele effect of qFT12.1 and revealed that the recessive allele of qFT12.1 resulted in delayed flowering. Evaluating the qFT12.1 near-isogenic lines (NILs) under different growth conditions showed that NILs with the wild soybean genotype always showed later flowering than those with the cultivated soybean genotype. qFT12.1 was delimited to a 2703-kb interval between the markers BARCSOYSSR_12_0220 and BARCSOYSSR_12_0368 on chromosome 12. qFT12.1 may be a new flowering time gene locus in soybean.     

A new <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> deletion mutant <Emphasis Type="Italic">apetala1-20</Emphasis>

A new deletion allele of the APETALA1 (AP1) gene encoding a type II MADS-box protein with the key role in the initiation of flowering and development of perianth organs has been identified in A. thaliana. The deletion of seven amino acids in the conserved region of the K domain in the ap1-20 mutant considerably delayed flowering and led to a less pronounced abnormality in the corolla development compared to the weak ap1-3 and intermediate ap1-6 alleles. At the same time, a considerable stamen reduction has been revealed in ap1-20 as distinct from ap1-3 and ap1-6 alleles. These data indicate that the K domain of AP1 can be crucial for the initiation of flowering and expression regulation of B-class genes controlling stamen development.     

Patterns of subspecies genetic diversity among oilseed <Emphasis Type="Italic">Brassica rapa</Emphasis> as revealed by agro-morphological traits and SSR markers

Brassica rapa (2n = 20, AA genome) is an important oil yielding species of the family Brassicaceae and characterized by wide range of genetic and morphological subtypes suitable for cultivation under diverse agro-climatic regions of India. In this study, genetic diversity among three subspecies of B. rapa including yellow sarson, toria and outlier brown sarson was estimated using various agro-morphological traits and simple sequence repeat (SSR) markers. Maximum variability was recorded for siliqua angle (Coefficient of variation = 30.9%), followed by seeds/siliqua (CV = 18.8%), leaf length (CV = 10%) and plant height (CV = 16.8%). Principal component analysis explained more than 50% of the total observed morphological variability for first two components. Of the 107 SSR markers tested, 80 generated reproducible, clear and distinct amplicons of which, 65 (81.25%) were found polymorphic. The number of alleles at each locus ranged from 2 to 7, with an average of 3.03 alleles per marker. A total of 197 alleles were detected at 65 SSR loci with average PIC value of 0.457 and a mean resolving power of 3.04. Neighbor-Joining cluster analysis based on morphological traits and SSR markers separately classified all the 28 genotypes into five major groups. The population structure analysis resulted into three sub-populations with certain extent of admixture among the earlier established taxonomic sub-groups. Twenty-three unique alleles were detected in thirteen B. rapa varieties. The clustering analysis and principal coordinate analysis outlined the genetic relationships among different varieties belonging to the three subspecies of B. rapa. Genetically diverse genotypes as illustrated by score plots and from the clustering patterns brought out the wide range of diversity present among B. rapa genotypes and the underlying options available for selecting parental genotypes for hybridization and developing high yielding cultivars suitable for Indian conditions.     

Development of SSR markers and identification of major quantitative trait loci controlling shelling percentage in cultivated peanut (<Emphasis Type="Italic">Arachis hypogaea</Emphasis> L.)

Key message

A total of 204,439 SSR markers were developed in diploid genomes, and 25 QTLs for shelling percentage were identified in a RIL population across 4 years including five consistent QTLs.

Abstract

Cultivated peanut (Arachis hypogaea L.) is an important grain legume providing edible oil and protein for human nutrition. Genome sequences of its diploid ancestors, Arachis duranensis and A. ipaensis, were reported, but their SSRs have not been well exploited and utilized hitherto. Shelling percentage is an important economic trait and its improvement has been one of the major objectives in peanut breeding programs. In this study, the genome sequences of A. duranensis and A. ipaensis were used to develop SSR markers, and a mapping population (Yuanza 9102 × Xuzhou 68-4) with 195 recombinant inbred lines was used to map QTLs controlling shelling percentage. The numbers of newly developed SSR markers were 84,383 and 120,056 in the A. duranensis and A. ipaensis genomes, respectively. Genotyping of the mapping population was conducted with both newly developed and previously reported markers. QTL analysis using the phenotyping data generated in Wuhan across four consecutive years and genotyping data of 830 mapped loci identified 25 QTLs with 4.46–17.01% of phenotypic variance explained in the four environments. Meta-analysis revealed five consistent QTLs that could be detected in at least two environments. Notably, the consistent QTL cqSPA09 was detected in all four environments and explained 10.47–17.01% of the phenotypic variance. The segregation in the progeny of a residual heterozygous line confirmed that the cpSPA09 locus had additive effect in increasing shelling percentage. These consistent and major QTL regions provide opportunity not only for further gene discovery, but also for the development of functional markers for breeding.

     

Inheritance and QTL mapping of resistance to gummy stem blight in cucumber stem

Gummy stem blight (GSB), a common disease of all major cucurbits, is caused by the fungus Didymella bryoniae. It results in serious losses in fruit production, which in cucumber can be up to 80% or more. Because the severity of the disease varies from season to season and also because of the harm to the environment caused by using pesticides to control the disease, the best method for overcoming GSB in cucumber is to develop more resistant cultivars by molecular breeding. There are no reports on molecular markers for use in breeding GSB resistance and no studies on chromosomal mapping of resistance. In this paper, a set of 160 F₉ recombinant inbred lines (RILs) were derived from the cross between the wild-type GSB-resistant cucumber accession PI 183967 and the cultivated GSB-susceptible accession 931. A total of 2112 pairs of SSR primers were used to study the inheritance of GSB resistance and to detect quantitative trait loci (QTLs) conferring resistance in the cucumber stem. Genetic analysis indicated that resistance to GSB in PI 183967 was quantitative and mainly governed by three pairs of additive epistatic major genes. Five QTLs, gsb-s1.1, gsb-s2.1, gsb-s6.1, gsb-s6.2, and gsb-s6.3, for resistance to GSB in cucumber stems were detected. The loci gsb-s1.1 and gsb-s2.1 with phenotypic variations of 8.7 and 6.7% were mapped to chromosomes (Chr.) 1 and 2, respectively. The loci gsb-s6.1, gsb-s6.2, and gsb-s6.3 were linked on Chr.6. Locus gsb-s6.2 accounted for the highest phenotypic variation of 22.7% and was flanked by markers SSR04083 and SSR02940 with genetic distances of 5.0 and 1.8 cM, respectively. There were 117 candidate genes predicted between SSR04083 and SSR02940, of which 14 were related to disease resistance.     

Development of a robust marker for <Emphasis Type="Italic">Psy-1</Emphasis> homoeologs and its application in improvement of yellow pigment content in durum wheat

Phytoene synthase-1 (Psy-1) homoeologs are associated with yellow pigment content (YPC) in endosperm of durum and bread wheat. In the present study, microsatellite variation in promoter region of Psy-A1 was identified in durum wheat and marker Psy-1SSR, targeting the microsatellite variation was developed which amplifies variation in Psy-A1 and Psy-B1 loci simultaneously. Psy-A1SSR was mapped within QYp.macs-7A, a major QTL for YPC identified earlier in PDW 233/Bhalegaon 4 population. Marker Psy-A1SSR was further validated in two different RIL populations and a set of 222 tetraploid wheat accessions including less cultivated tetraploid wheat species. Eight alleles of Psy-A1SSR were identified in 222 wheat accessions, while seven alleles were observed for Psy-B1SSR. Variation at Psy-A1SSR showed significant association with YPC, whereas no association was observed with Psy-B1SSR. Marker-assisted introgression of Psy-A1SSRe allele from PDW 233, to durum wheat cultivars MACS 3125 and HI 8498 resulted in improvement of YPC. Backcrossed BC₃F_2:4 and BC₂F_2:3 lines selected using Psy-A1SSR showed 89 to 98% gain in YPC over recurrent parents indicating robustness of marker. The marker can thus be utilized in marker-assisted improvement of YPC in durum wheat cultivars.     

Genetic characterization of <Emphasis Type="Italic">Vitis</Emphasis> germplasm collected from the southwestern US and Mexico to expedite Pierce’s disease-resistance breeding

Pierce’s disease (PD) limits the cultivation of Vitis vinifera grape cultivars in California, across the southern United States and into South America. Resistance has been well characterized in V. arizonica, and one resistance locus has been identified (PdR1). However, resistance is poorly characterized in most other grape species. We tested a wide range of Vitis species from the southwestern United States for resistance to PD and used nuclear and chloroplast markers to phenotypically and genetically select a diverse set of resistant accessions. Chloroplast SSR markers identified 11 maternal lineage lines within the set of 17 (14 new and three previously identified) PD resistant accessions. A total of 19 breeding populations (F1 and pseudo-BC1) were developed with the 14 PD resistant accessions, and a total of 705 seedlings were analyzed for PD resistance. Using a limited mapping approach, 12 SSR markers, linked to the PdR1 locus, were used to genotype the breeding populations and phenotypic data were analyzed. Nine accessions had a major resistance quantitative trait locus (QTL) within the genomic region containing PdR1. The phenotypic data for these three resistant accessions, ANU67, b41-13, and T03-16, did not associate with PdR1 linked markers, indicating that their resistance is located in other regions of the genome. These three accessions were identified as candidates for use in the development of framework maps with larger populations capable of detecting additional and unique loci for PD resistance breeding and the stacking of PD resistance genes.     

QTL analysis and the development of closely linked markers for days to flowering in spring oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

Days to flowering (DTF) is an important trait impacting cultivar performance in oilseed rape (Brassica napus L.), but the interaction of all loci controlling this trait in spring-type oilseed rape is not fully understood. We identified quantitative trait loci (QTL) for variation in DTF in a doubled haploid (DH) population from the Qinghai–Tibet Plateau that includes 217 lines derived from a cross between spring-type oilseed rape (B. napus L.) line No. 5246 and line No. 4512, the latter of which is responsive to the effective accumulated temperature (EAT). A linkage map was constructed for the DH population, using 202 SSR and 293 AFLP markers. At least 22 DTF QTL were found in multiple environments. Four major QTL were located on linkage groups A7, C2, C8 and C8. Among these QTL, cqDTFA7a and cqDTFC2a were identified in five environments and individually explained 10.4 and 23.0 % of the trait variation, respectively. cqDTFC8, a major QTL observed in spring environments, and a unique winter environment QTL, qDTFC8-3, were identified; these QTL explained 10.0 and 46.5 % of the phenotypic variation, respectively. Minor QTL (for example, cqDTFC2c) and epistatic interactions seemed evident in this population. Two closely linked SSR markers for cqDTFA7a and cqDTFC8 were developed (G1803 and S034). BnAP1, a B. napus gene with homology to Arabidopsis thaliana that was identified as a cqDTFA7a candidate gene, played a major role in this study. The allelic effects of the major and minor QTL on DTF were further validated in the DH population and in 93 breeding genotypes.     

Phylogeographic structure of <Emphasis Type="Italic">Spondias tuberosa</Emphasis> Arruda Câmara (Anacardiaceae): seasonally dry tropical forest as a large and continuous refuge

Spondias tuberosa occurs in the Caatinga domain (seasonally dry tropical forest biome) of north-eastern Brazil, a large biome with ecogeographic regions that may have modelled the population structure of the species. Here we studied the phylogeographic pattern of S. tuberosa using sequences of the accD-psaI plastid region and six SSR markers in individuals distributed across 20 localities. The results for accD-psaI demonstrated nine haplotypes: some of which were exclusive to Caatinga ecoregions, whereas others were found in all localities. Spatial analysis of molecular variance revealed two groups (F_ct?=?0.34, P?<?0.0039) with 33.91% variation between them. The SSR analyses displayed 2–5 alleles at each locus, some of which were unique to certain localities. As in the accD-psaI region, the population structure obtained using SSR markers fell into two groups: (1) a large group containing the majority of the geographic region of Caatinga and (2) a small group near the Atlantic forest. We demonstrate the population structure of S. tuberosa, identifying the Caatinga as large, continuous refuge and the region near the interface between the Caatinga and the Atlantic forest as second refuge.