Parallel domestication, convergent evolution and duplicated gene recruitment in allopolyploid cotton

A putative advantage of allopolyploidy is the possibility of differential selection of duplicated (homeologous) genes originating from two different progenitor genomes. In this note we explore this hypothesis using a high throughput, SNP-specific microarray technology applied to seed trichomes (cotton) harvested from three developmental time points in wild and modern accessions of two independently domesticated cotton species, Gossypium hirsutum and G. barbadense. We show that homeolog expression ratios are dynamic both developmentally and over the several-thousand-year period encompassed by domestication and crop improvement, and that domestication increased the modulation of homeologous gene expression. In both species, D-genome expression was preferentially enhanced under human selection pressure, but for nonoverlapping sets of genes for the two independent domestication events. Our data suggest that human selection may have operated on different components of the fiber developmental genetic program in G. hirsutum and G. barbadense, leading to convergent rather than parallel genetic alterations and resulting morphology.     

A genetic bottleneck in the ’evolution under domestication’ of upland cotton Gossypium hirsutum L. examined using DNA fingerprinting

Reliable information about the evolutionary and genetic relationships of various germplasm resources is essential to the establishment of rational strategies for crop improvement. We used AFLPs to study the genetic relationships of 43 cultivars of Gossypium hirsutum representative of the genomic composition of modern ’Upland’ cotton. The study also included representatives of the related tetraploid species Gossypium barbadense, as well as the diploid species Gossypium raimondii, Gossypium incanum, Gossypium herbaceum and Gossypium arboreum. We tested 20 primer combinations that resulted in a total of 3,178 fragments. At the species level, and above, genetic similarities based on AFLPs were in agreement with the known taxonomic relationships. Similarity indices ranged from 0.25 to 0.99. Representatives of the G. hirsutum germplasm resources utilized in North America, including secondary accessions collected by breeders in Central America (’Acala’, ’Tuxtla’, ’Kekchi’) and the southwestern US (’Hopi Moencopi’), formed a single cluster with exceedingly limited genetic diversity (with many pairwise similarity indices >0.96) We concluded that these accessions were derived from the same genetic pool. The early maturing or ’latifolium’ or ’Mexican Highlands’ cultigens from which these cultivars were derived appear to have had extremely limited genetic diversity, perhaps as a result of a severe genetic bottleneck resulting from the selection pressures of domestication. Outside of the major G. hirsutum cluster, well-supported phylogenies were inferred. Inside this cluster, phylogenies were obscured by limited diversity, reticulation and lineage sorting. The implications of these findings for cotton improvement are discussed. Received: 23 May 2000 / Accepted: 23 January 2001     

Genetic fingerprinting of Australian cotton cultivars with RAPD markers.

RAPD (random amplified polymorphic DNA) markers generated by 30 random decamer primers were used to fingerprint 12 released cultivars and a breeding line of Gossypium hirsutum and 1 cultivar of G. barbadense presently under cultivation in Australia. Among a total of 453 developed markers, 69 (15.2%) were only present (unique) in the G. barbadense cultivar Pima S-7. Of the remaining markers, 128 (33.3%) were fixed in all 13 G. hirsutum cultivars. In pairwise comparisons of the degree of band sharing, nine closely-related cultivars showed 92.1-98.9% genetic similarity. Cluster analysis of genetic distance estimates between each of the cultivars revealed phylogenetic relationships in broad agreement with the known lineage of the cultivars. Ten of the G. hirsutum cultivars can be characterized individually based upon cultivar-specific RAPD markers, thus making it possible to differentiate closely related cultivars by molecular markers.     

Assessing genetic diversity in <Emphasis Type="Italic">Gossypium arboreum</Emphasis> L. cultivars using genomic and EST-derived microsatellites

The cultivated diploid, Gossypium arboreum L., (A genome) is an invaluable genetic resource for improving modern tetraploid cotton (G. hirsutum L. and G. barbadense L.) cultivars. The objective of this research is to select a set of informative and robust microsatellites for studying genetic relationships among accessions of geographically diverse G. arboreum cultivars. From more than 1,500 previously developed simple sequence repeat (SSR) markers, 115 genomic (BNL) and EST-derived (MUCS and MUSS) markers were used to evaluate the allelic diversity of a core panel of G. arboreum accessions. These SSR data enabled advanced genome analyses. A set of 25 SSRs were selected based both upon their high level of informativeness (PIC ≥ 0.50) and the production of clear PCR bands on agarose gels. Subsequently, 96 accessions representing a wide spectrum of diversity of G. arboreum cultivars were analyzed with these markers. The 25 SSR loci revealed 75 allelic variants (polymorphisms) ranging from 2 to 4 alleles per locus. The Neighborjoining (NJ) method, based on genetic dissimilarities, revealed that cultivars from geographically adjacent countries tend to cluster together. Outcomes of this research should be useful in decreasing redundancy of effort and in constructing a core collection of G. arboreum, important for efficient use of this genetic resource in cotton breeding.     

Resequencing core accessions of a pedigree identifies derivation of genomic segments and key agronomic trait loci during cotton improvement

Upland cotton (Gossypium hirsutum) is the world's largest source of natural fibre and dominates the global textile industry. Hybrid cotton varieties exhibit strong heterosis that confers high fibre yields, yet the genome‐wide effects of artificial selection that have influenced Upland cotton during its breeding history are poorly understood. Here, we resequenced Upland cotton genomes and constructed a variation map of an intact breeding pedigree comprising seven elite and 19 backbone parents. Compared to wild accessions, the 26 pedigree accessions underwent strong artificial selection during domestication that has resulted in reduced genetic diversity but stronger linkage disequilibrium and higher extents of selective sweeps. In contrast to the backbone parents, the elite parents have acquired significantly improved agronomic traits, with an especially pronounced increase in the lint percentage. Notably, identify by descent (IBD) tracking revealed that the elite parents inherited abundant beneficial trait segments and loci from the backbone parents and our combined analyses led to the identification of a core genomic segment which was inherited in the elite lines from the parents Zhong 7263 and Ejing 1 and that was strongly associated with lint percentage. Additionally, SNP correlation analysis of this core segment showed that a non‐synonymous SNP (A‐to‐G) site in a gene encoding the cell wall‐associated receptor‐like kinase 3 (GhWAKL3) protein was highly correlated with increased lint percentage. Our results substantially increase the valuable genomics resources available for future genetic and functional genomics studies of cotton and reveal insights that will facilitate yield increases in the molecular breeding of cotton.     

Reevaluating the origin of domesticated cotton (Gossypium hirsutum; Malvaceae) using nuclear restriction fragment length polymorphisms (RFLPs)

The origin of domestication in Mesoamerican G. hirsutum populations is obscured by several factors, including the absence of a clearly identified wild progenitor, a complex population genetic structure, and many centuries of human-mediated dispersal and gene flow. Phenetic and phylogenetic analyses of allelic variation at 205 restriction fragment length polymorphism (RFLP) loci were conducted in an effort to unravel this complicated history. The RFLP data, in conjunction with previously published molecular, morphological, and anthropological information, suggest that coastal Yucatan populations are truly wild rather than reestablished feral derivatives. The geographical proximity of these wild coastal populations to agronomically primitive forms of G. hirsutum implicates the Yucatan peninsula as the primary site for the earliest stages of domestication. Agronomically advanced cultigens developed in southern Mexico and Guatemala appear to have been derived from introduced Yucatan peninsular forms, thereby creating the secondary center of diversity that has traditionally been interpreted as the geographical point of origin of domesticated G. hirsutum. The gene pool of modern, improved (Upland) cultivars derives from Mexican highland populations that, in turn, trace their origins to southern Mexico and Guatemala. Gossypium hirsutum is the first tetraploid perennial surveyed for RFLP variation. Levels of RFLP variation in G. hirsutum (H_T = 0.048, A = 1.24, and P = 22%) are low relative to other plant taxa, and, in contrast to the few comparable studies, levels of allozyme variation are higher than levels of RFLP variation. Despite assaying 205 loci, only six of the 23 Upland cultivars were found to have unique multilocus genotypes.     

Population genomics reveals a fine‐scale recombination landscape for genetic improvement of cotton

Recombination breaks up ancestral linkage disequilibrium, creates combinations of alleles, affects the efficiency of natural selection, and plays a major role in crop domestication and improvement. However, there is little knowledge regarding the variation in the population‐scaled recombination rate in cotton. We constructed recombination maps and characterized the difference in the genomic landscape of the population‐scaled recombination rate between Gossypium hirsutum and G. arboreum and sub‐genomes based on the 381 sequenced G. hirsutum and 215 G. arboreum accessions. Comparative genomics identified large structural variations and syntenic genes in the recombination regions, suggesting that recombination was related to structural variation and occurred preferentially in the distal chromosomal regions. Correlation analysis indicated that recombination was only slightly affected by geographical distribution and breeding period. A genome‐wide association study (GWAS) was performed with 15 agronomic traits using 267 cotton accessions and identified 163 quantitative trait loci (QTL) and an important candidate gene (Ghir_COL2) for early maturity traits. Comparative analysis of recombination and a GWAS revealed that the QTL of fibre quality traits tended to be more common in high‐recombination regions than were those of yield and early maturity traits. These results provide insights into the population‐scaled recombination landscape, suggesting that recombination contributed to the domestication and improvement of cotton, which provides a useful reference for studying recombination in other species.     

Allozyme evidence for the origin and diversification of Gossypium barbadense L.

Summary Gossypium barbadense L. is a commercially important cotton species of tropical South American origin presently grownin many regions of the world. The species is morphologically diverse, consisting of a wide range of wild (or feral), commensal, landrace, and highly improvedcommercial forms. We performed allozyme analysis on 153 accessions representing the spectrum of G. barbadense diversityto ascertain the geographic origin of the species, its patterns of diffusion subsequent to domestication, and to reveal infraspecific relationships. Levels ofgenetic variation in G. barbadense are moderate. Of 59 loci scored, 24 were polymorphic, with a mean number of alleles perlocus of 1.69 and an average panmictic heterozygosity of 0.062. Principal component analysis revealed geographic clustering of accessions into six relativelydiscrete regions. Gene frequencies at many loci are significantly heterogeneous among these regions, with an average G _STof 0.272. Northwestern South America contains the greatest genetic variability; we suggest that this region is the ancestral home of the species. The data indicate separate diffusion pathways from this region into Argentina-Paraguay and into eastern and northern South America east of the Andes. Caribbean Island and Central American forms appear to be derived from the latter. These diffusion pathways are in accordance with morphological evidence and historical record. In contrast to expectations based on geographic proximity, Pacific Island forms have their closest affinity to accessions from eastern South America. Advanced cultivated stocks seem largely derived from western Andean material, but also contain introgressed G. hirsutum germ plasm. Introgression was relatively high (22%–50% of accessions) in commercial stocks and in forms from Argentina-Paraguay and various Pacific Islands, but was conspicuously low or absent in material from Central America and the Caribbean, where commensal and commercial forms of both species are sympatric.     

Genomic divergence in cotton germplasm related to maturity and heterosis

Commercial varieties of upland cotton(Gossypium hirsutum) have undergone extensive breeding for agronomic traits, such as fiber quality, disease resistance,and yield. Cotton breeding programs have widely used Chinese upland cotton source germplasm(CUCSG) with excellent agronomic traits. A better understanding of the genetic diversity and genomic characteristics of these accessions could accelerate the identification of desirable alleles. Here, we analyzed 10,522 high-quality singlenucleotide polymorphisms(SNP) with the CottonSNP63 K microarray in 137 cotton accessions(including 12 hybrids of upland cotton). These data were used to investigate the genetic diversity, population structure,and genomic characteristics of each population and the contribution of these loci to heterosis. Three subgroups were identified, in agreement with their knownpedigrees, geographical distributions, and times since introduction. For each group, we identified lineagespecific genomic divergence regions, which potentially harbor key alleles that determine the characteristics of each group, such as early maturity-related loci. Investigation of the distribution of heterozygous loci, among 12 commercial cotton hybrids, revealed a potential role for these regions in heterosis. Our study provides insight into the population structure of upland cotton germplasm. Furthermore, the overlap between lineagespecific regions and heterozygous loci, in the high-yield hybrids, suggests a role for these regions in cotton heterosis.     

Frequency and fidelity of alien chromosome transmission in Gossypium hexaploid bridging populations.

The Australian diploid Gossypium species possess traits of potential agronomical value, such as gossypol-free seeds and Fusarium wilt resistance. However, they belong to the tertiary germplasm pool, which is the most difficult group of species from which to introgress genes into G. barbadense L. and G. hirsutum L. Interspecific triploid hybrids can be generated but they are sterile. The sterility barrier can be overcome using synthetic polyploids as introgression bridges, but whether there is sufficient homoeologous chromosome interaction at meiosis to allow recombination is still an open question. To ascertain, genetically, observable levels of homoeologous introgression, 2 synthetic hexaploid lines (2x G. hirsutum x G. australe and 2x G. hirsutum x G. sturtianum) were crossed to G. hirsutum to generate pentaploid F1 plants that, in turn, were backcrossed to G. hirsutum to generate BC1 and BC2 multiple alien chromosome addition lines (MACALs). Gossypium australe F. Muell. and G. sturtianum Willis chromosome-specific markers were used to track the frequency and fidelity of chromosome transmission to the BC1 and BC2 MACALs. The chromosomal location of the AFLP markers was determined by their distribution among the MACALs and confirmed in parental F2 families. Roughly half the available chromosomes were transmitted to the G. hirsutum x G. australe (54%) and G. hirsutum x G. sturtianum (52%) BC1 MACALs. The BC2 MACAL families again inherited about half of the available chromosomes. There were, however, notable exceptions for specific chromosomes. Some chromosomes were preferentially eliminated, while others were preferentially transmitted. Consistent with the genomic stability of Gossypium synthetic polyploids, the de novo loss or gain of AFLP fragments was rarely observed. While restructuring of the donor G. australe and G. sturtianum chromosomes was observed, this is more likely the result of chromatin loss, and no clear cases of introgression of donor chromatin into the recipient G. hirsutum genome were observed.     

Low levels of nucleotide diversity at homoeologous Adh loci in allotetraploid cotton (Gossypium L.).

Levels of genetic diversity within and among populations and species are shaped by both external (population-level) and internal (genomic and genic) evolutionary forces. To address the effect of internal pressures, we estimated nucleotide diversity for a pair of homoeologous Adh loci in an allotetraploid species, Gossypium hirsutum. These data represent the first such estimates for a pair of homoeologous nuclear loci in plants. Estimates of nucleotide diversity for AdhA in Gossypium are lower than those for any plant nuclear gene yet described. This low diversity appears to reflect primarily a history of repeated, severe genetic bottlenecks associated with both speciation and recent domestication, supplemented by an unusually slow nucleotide substitution rate and an autogamous breeding system. While not statistically supportable, the sum of the observations also suggest differential evolutionary dynamics at each of the homoeologous loci.     

Towards new sources of resistance to the currant-lettuce aphid (<Emphasis Type="Italic">Nasonovia ribisnigri</Emphasis>)

Domesticated lettuce varieties encompass much morphological variation across a range of crop type groups, with large collections of cultivars and landrace accessions maintained in genebanks. Additional variation not captured during domestication, present in ancestral wild relatives, represents a potentially rich source of alleles that can deliver to sustainable crop production. However, these large collections are difficult and costly to screen for many agronomically important traits. In this paper, we describe the generation of a diversity collection of 96 lettuce and wild species accessions that are amenable to routine phenotypic analysis and their genotypic characterization with a panel of 682 newly developed expressed sequence tag (EST)-linked KASP? single nucleotide polymorphism (SNP) markers that are anchored to the draft Lactuca sativa genome assembly. To exemplify the utility of these resources, we screened the collection for putative sources of resistance to currant-lettuce aphid (Nasonovia ribisnigri) and carried out association analyses to look for potential SNPs linked to resistance.     

Analysis of xyloglucan endotransglycosylase/hydrolase (XTH) genes from allotetraploid (Gossypium hirsutum) cotton and its diploid progenitors expressed during fiber elongation

Multiple cellular pathways have been shown to be involved during fiber initiation and elongation stages in the cultivated allotetraploid cotton (Gossypium hirsutum). The cell wall enzymes xyloglucan endotransglycosylase/hydrolases (XTH) have been reported to be associated with the biosynthesis of the cell wall and the growth of cotton fibers, probably regulating the plasticity of the primary cell wall. Among various cotton fiber cDNAs found to be preferentially expressed in cotton fibers, a xyloglucan endotransglycosylase (XTH) cDNA was significantly up-regulated during the elongation stage of cotton fiber development. In the present study, we isolated and characterized genomic clones encoding cotton XTH from cultivated cotton (Gossypium hirsutum) and its diploid progenitors (Gossypium arboreum and Gossypium raimondii), designated GhXTH1-1, GhXTH1-2, GaXTH1 and GrXTH, respectively. In addition, we isolated and characterized, by in silico methods, the putative promoter of XTH1 from Gossypium hirsutum. Sequence analysis revealed more than 50% homology to XTH's at the protein level. DNA gel blot hybridization indicated that at least two copies of GhXTH1 are present in Gossypium hirsutum whereas the diploid progenitor species Gossypium arboreum and Gossypium raimondii has only a single copy. Quantitative real-time PCR and high-resolution melting experiments indicated that in Gossypium hirsutum cultivars, in cotton fibers during early stages of fiber elongation specifically expressing only the GhXTH1-1 gene and expression levels of GhXTH1-1 in fibers varies among cultivars differing in fiber percentage and fiber length.     

Esterase Isozyme analyses of the Species and Their Varieties of the Genus Gossypium

The isozyme make up of esterases of the seeds from fifteen species and twenty-three cultivar of Gossypium was analyzed by isoelectrofocusing. The experimental results are summarized as follows: 1. Differeces were observed in the number of esterase isozyme bands among the species of different genome groups. The cultivated species, G. hirsutum (AD)1 gave rise to 46 isozyme bands, the most among the species of the genus Gossypium. G. barba- dense (AD)2, G. arboreum (A2) and G. herbaceum (A1) gave rise to 42, 40 and 38 bands, respectively. In wild species, G. australe (C3) had 20 esterase bands, the least in all species of Gossypium. The bands given rise from other wild species ranged from 26 to 40. 2. Each species of every genome groups had its marker bands. The results were in agreement with the traditional classification and provided some biochemical evidence for modern classification of Gossypium. 3. It was clear that all cotton species of different genome groups contain 5 main isozyme bands, viz. PI=3.85, 4.61, 5.48, 5.73 and 5.91 in the zymograms. In other words, these zymograms are common characters of Gossypium. 4. The esterase of 23 cultivers in four cultivated species studied showed that no variation in isozyme patterns existed within one species, except the disease-resistant variety Hea-7124 which differs from other 4 cultivars of G. barbadense.     

Infraspecific DNA methylation polymorphism in cotton (Gossypium hirsutum L.)

Cytosine methylation is important in the epigenetic regulation of gene expression and development in plants and has been implicated in silencing duplicate genes after polyploid formation in several plant groups. Relatively little information exists, however, on levels and patterns of methylation polymorphism (MP) at homologous loci within species. Here we explored the levels and patterns of methylation-polymorphism diversity at CCGG sites within allotetraploid cotton, Gossypium hirsutum, using a methylation-sensitive amplified fragment length polymorphism screen and a selected set of 20 G. hirsutum accessions for which we have information on genetic polymorphism levels and relationships. Methylation and MP exist at high levels within G. hirsutum: of 150 HpaII/MspI sites surveyed, 48 were methylated at the inner cytosine (32%) and 32 of these were polymorphic (67%). Both these values are higher than comparable measures of genetic diversity using restriction fragment length polymorphisms. The high percentage of methylation-polymorphic sites and potential relationship to gene expression underscore the potential significance of MP within and among populations. We speculate that biased correlation of methylation-polymorphic sites and genes in cotton may be a consequence of polyploidy and the attendant doubling of all genes.     

Simple sequence repeat (SSR) analysis for assessment of genetic variability in apricot germplasm

Thirty SSR primer combinations, developed from peach SSR-enriched genomic libraries and BAC libraries of peach [ Prunus persica (L.) Batsch.], were tested for cross amplification with 74 apricot ( Prunus armeniaca L.) germplasm accessions. Twelve primer pairs amplified 14 polymorphic SSR loci useful for discriminating most apricot cultivars, as well as for investigating patterns of variation in apricot germplasm. Levels of polymorphism were higher than the levels described using other codominant marker systems (i.e., isozymes, RFLP markers). Overall, 107 alleles were identified, and all but 11 accessions were unambiguously discriminated. Genetic differentiation of native germplasm into traditional ecogeographical groups was low, with a high level of genetic identity (> 0.75) between the groups. However, neighbor joining cluster analysis of marker distances between cultivars reflected the complex history of apricot domestication, producing groupings not evidently based on the geographical origin of the cultivars. Distant positioning of Chinese cultivars on UPGMA and neighbor joining dendrograms supports the authors' consideration of Chinese apricots as subspecies, Prunus armeniaca var. ansu Maxim., rather than a separate species.     

Inheritance of Resistance to Meloidoygne incognita in Primitive Cotton Accessions from Mexico

Few sources of resistance to root-knot nematodes (Meloidogyne incognita) in upland cotton (Gossypium hirsutum) have been utilized to develop resistant cultivars, making this resistance vulnerable to virulence in the pathogen population. The objectives of this study were to determine the inheritance of resistance in five primitive accessions of G. hirsutum (TX1174, TX1440, TX2076, TX2079, and TX2107) and to determine allelic relations with the genes for resistance in the genotypes Clevewilt-6 (CW) and Wild Mexico Jack Jones (WMJJ). A half-diallel experimental design was used to create 28 populations from crosses among these seven sources of resistance and the susceptible cultivar DeltaPine 90 (DP90). Resistance to M. incognita was measured as eggs per g roots in the parents, F(1) and F(2) generations of each cross. The resistance in CW and WMJJ was inherited as recessive traits, as reported previously for CW, whereas the resistance in the TX accessions was inherited as a dominant trait. Chi square analysis of segregation of resistance in the F(2) was used to estimate the numbers of genes that conditioned resistance. Resistance in CW and WMJJ appeared to be a multigenic trait whereas the resistance in the TX accessions best fit either a one or two gene model. The TX accessions were screened with nine SSR markers linked to resistance loci in other cotton genotypes. The TX accessions lacked the allele amplified by SSR marker CR316 and linked to resistance in CW and other resistant genotypes derived from this source. Four of five TX genotypes lacked the amplification products from the marker BNL1231 that is also associated with the resistant allele on Chromosome 11 in WMJJ, CW, NemX, M120 RNR and Auburn 634 RNR. However, all five TX genotypes produced the same amplification products from three SSR markers linked to the resistant allele on Chromosome 14 in M120 RNR and M240 RNR. The TX accessions have unique resistance genes that are likely to be useful in efforts to develop resistant cotton cultivars with increased durability.     

Comparative effect of allopolyploidy on transposable element composition and gene expression between Gossypium hirsutum and its two diploid progenitors

An allopolyploidization event formed allotetraploid Gossypium species from an A-genome diploid species and a D-genome diploid species. To explore the responses of transposable elements(TEs) to allopolyploidy, we assembled parallel TE datasets from G. hirsutum, G. arboreum and G. raimondii and analyzed the TE types and the effects of TEs on orthologous gene expression in the three Gossypium genomes.Gypsy was the most abundant TE type and most TEs were located $500 bp from genes in all three genomes. In G. hirsutum, 35.6% of genes harbored TE insertions, whereas insertions were more frequent in G. arboreum and G. raimondii. G. hirsutum had the highest proportion of uniquely matching 24-nt small interfering RNAs(siRNAs) that targeted TEs. TEs,particularly those targeted by 24-nt siRNAs, were associated with reduced gene expression, but the effect of TEs on orthologous gene expression varied substantially among species. Orthologous gene expression levels in G. hirsutum were intermediate between those of G. arboreum and G. raimondii, which did not experience TE expansion or reduction resulting from allopolyploidization. This study underscores the diversity of TEs co-opted by host genes and provides insights into the roles of TEs in regulating gene expression in Gossypium.