Sterility Barriers Research Articles

Reinstatement of the fission yeast species Schizosaccharomyces versatilis Wickerham et Duprat, a sibling species of Schizosaccharomyces japonicus.

Schizosaccharomyces japonicus Yukawa et Maki (1931) and Schizosaccharomyces versatilis Wickerham et Duprat (1945) have been treated as varieties of S. japonicus or as conspecific, based on various approaches including mating trials and nDNA/nDNA optical reassociation studies. However, the type strains of S. japonicus and S. versatilis differ by five substitutions (99.15% identity) and one 1-bp indel in the sequences of the D1/D2 domain of the 26S rRNA gene, and 23 substitutions (96.3% identity) and 31-bp indels in the sequences of internal transcribed spacer (ITS) of rRNA, suggesting that they may not be conspecific. To reassess their taxonomic status, we conducted mating trials and whole-genome analyses. Mating trials using the type strains showed a strong but incomplete prezygotic sterility barrier, yielding interspecies mating products at two orders of magnitude lower efficiency than intraspecies matings. These mating products, which were exclusively allodiploid hybrids, were unable to undergo the haplontic life cycle of the parents. We generated chromosome-level gap-less genome assemblies for both type strains. Whole genome sequences yielded an average nucleotide identity (ANI) of 86.4%, indicating clear separation of S. japonicus and S. versatilis. Based on these findings, we propose the reinstatement of S. versatilis as a distinct species (holotype strain: CBS 103T and ex-types: NRRL Y-1026, NBRC 1607, ATCC 9987, PYCC 7100; Mycobank no.: 847838).

Autotetraploid Rice Hybrids: Overcoming Sterility Barriers for Enhanced Heterosis

Autotetraploid Rice Hybrids: Overcoming Sterility Barriers for Enhanced Heterosis

Synthetic two-species allodiploid and three-species allotetraploid Saccharomyces hybrids with euploid (complete) parental subgenomes

Combination of the genomes of Saccharomyces species has great potential for the construction of new industrial strains as well as for the study of the process of speciation. However, these species are reproductively isolated by a double sterility barrier. The first barrier is mainly due to the failure of the chromosomes to pair in allodiploid meiosis. The second barrier ensures that the hybrid remains sterile even after genome duplication, an event that can restore fertility in plant interspecies hybrids. The latter is attributable to the autodiploidisation of the allotetraploid meiosis that results in sterile allodiploid spores (return to the first barrier). Occasionally, mating-competent alloaneuploid spores arise by malsegregation of MAT-carrying chromosomes. These can mate with cells of a third species resulting in aneuploid zygotes having at least one incomplete subgenome. Here we report on the construction of euploid three-species hybrids by making use of “rare mating” between a sterile S. kudriavzevii x S. uvarum allodiploid hybrid and a diploid S. cerevisiae strain. The hybrids have allotetraploid 2nScnSk nSu genomes consisting of complete sets of parental chromosomes. This is the first report on the production of euploid three-species Saccharomyces hybrids by natural mating, without genetic manipulation. The hybrids provide possibilities for studying the interactions of three allospecific genomes and their orthologous genes present in the same cell.

Diversity and Postzygotic Evolution of the Mitochondrial Genome in Hybrids of Saccharomyces Species Isolated by Double Sterility Barrier.

Eukaryotic species are reproductively isolated by sterility barriers that prevent interspecies fertilization (prezygotic sterility barrier) or the fertilization results in infertile offspring (postzygotic sterility barrier). The Saccharomyces species are isolated by postzygotic sterility barriers. Their allodiploid hybrids form no viable gametes (ascospores) and the viable ascospores of the allotetraploids cannot fertilize (conjugate). Our previous work revealed that this mechanism of reproductive isolation differs from those operating in plants and animals and we designated it double sterility barrier (the failure of homeologous chromosomes to pair and the repression of mating by mating-type heterozygosity). Other studies implicated nucleo-mitochondrial incompatibilities in the sterility of the Saccharomyces hybrids, a mechanism assumed to play a central role in the reproductive isolation of animal species. In this project the mitochondrial genomes of 50 cevarum (S. cerevisiae × S. uvarum) hybrids were analyzed. 62% had S. cerevisiae mitotypes, 4% had S. uvarum mitotypes, and 34% had recombinant mitotypes. All but one hybrid formed viable spores indicating that they had genomes larger than allodiploid. Most of these spores were sterile (no sporulation in the clone of vegetative descendants; a feature characteristic of allodiploids). But regardless of their mitotypes, most hybrids could also form fertile alloaneuploid spore clones at low frequencies upon the loss of the MAT-carrying chromosome of the S. uvarum subgenome during meiosis. Hence, the cevarum alloploid nuclear genome is compatible with both parental mitochondrial genomes as well as with their recombinants, and the sterility of the hybrids is maintained by the double sterility barrier (determined in the nuclear genome) rather than by nucleo-mitochondrial incompatibilities. During allotetraploid sporulation both the nuclear and the mitochondrial genomes of the hybrids could segregate but no correlation was observed between the sterility or the fertility of the spore clones and their mitotypes. Nucleo-mitochondrial incompatibility was manifested as respiration deficiency in certain meiotic segregants. As respiration is required for meiosis-sporulation but not for fertilization (conjugation), these segregants were deficient only in sporulation. Thus, the nucleo-mitochondrial incompatibility affects the sexual processes only indirectly through the inactivation of respiration and causes only partial sterility in certain segregant spore clones.

MAT heterozygosity and the second sterility barrier in the reproductive isolation of Saccharomyces species

The genetic analysis of large numbers of Saccharomyces cerevisiae × S. uvarum (“cevarum”) and S. kudriavzevii × S. uvarum (“kudvarum”) hybrids in our previous studies revealed that these species are isolated by a postzygotic double-sterility barrier. We proposed a model in which the first barrier is due to the abruption of the meiotic process by the failure of the chromosomes of the subgenomes to pair (and recombine) in meiosis and the second barrier is assumed to be the result of the suppression of mating by allospecific MAT heterozygosity. While the former is analogous to the major mechanism of postzygotic reproductive isolation in plants and animals, the latter seems to be Saccharomyces specific. To bolster the assumed involvement of MAT in the second sterility barrier, we produced synthetic alloploid two-species cevarum and kudvarum hybrids with homo- and heterothallic backgrounds as well as three-species S. cerevisiae × S. kudvarum × S. uvarum (“cekudvarum”) hybrids by mass-mating and examined their MAT loci using species- and cassette-specific primer pairs. We found that the allospecific MAT heterozygosity repressed MAT switching and mating in the hybrids and in the viable but sterile spores produced by the cevarum hybrids that had increased (allotetraploid) genomes. The loss of heterozygosity by meiotic malsegregation of MAT-carrying chromosomes in the latter hybrids broke down the sterility barrier. The resulting spores nullisomic for the S. uvarum chromosome produced vegetative cells capable of MAT switching and conjugation, opening the way for GARMe (Genome Autoreduction in Meiosis), the process that leads to chimeric genomes.

Genetic variability among autotetraploid populations of banana plants derived from wild diploids through chromosome doubling using SSR and molecular markers based on retrotransposons

The development of banana cultivars resistant to diseases is the most viable way to minimize crop damages. However, this strategy is limited by the low fertility observed among commercial triploid cultivars, especially those of the Cavendish subgroup. Induced polyploidy has the potential to overcome sterility barriers and allows the development of triploid genotypes with fruit sensory profiles aligned with market demands. This work characterized agronomically five autotetraploid plant populations derived from diploids treated with colchicine. The experiment was conducted using an augmented block design, and 13 agronomic characteristics were evaluated during two production cycles. The sum of ranks proposed by Mulamba and Mock allowed the genotypes Pisang Lilin 8, Malbut 10, Niyarma Yik 2, Pisang Mas 3, and Thong Dok Mak 2 to be selected, which were superior for most of the agronomic characteristics evaluated. SSR and IRAP molecular markers detected variability in the autotetraploid populations, and the analysis of molecular variance supports that approximately 78% of identified variability was contained within the populations studied. Based on the results, it is inferred that the promising autotetraploids can be crossed with diploids to develop secondary triploid hybrids.

Yeast two- and three-species hybrids and high-sugar fermentation.

SummaryThe dominating strains of most sugar‐based natural and industrial fermentations either belong to Saccharomyces cerevisiae and Saccharomyces uvarum or are their chimeric derivatives. Osmotolerance is an essential trait of these strains for industrial applications in which typically high concentrations of sugars are used. As the ability of the cells to cope with the hyperosmotic stress is under polygenic control, significant improvement can be expected from concerted modification of the activity of multiple genes or from creating new genomes harbouring positive alleles of strains of two or more species. In this review, the application of the methods of intergeneric and interspecies hybridization to fitness improvement of strains used under high‐sugar fermentation conditions is discussed. By protoplast fusion and heterospecific mating, hybrids can be obtained that outperform the parental strains in certain technological parameters including osmotolerance. Spontaneous postzygotic genome evolution during mitotic propagation (GARMi) and meiosis after the breakdown of the sterility barrier by loss of MAT heterozygosity (GARMe) can be exploited for further improvement. Both processes result in derivatives of chimeric genomes, some of which can be superior both to the parental strains and to the hybrid. Three‐species hybridization represents further perspectives.

Interspecies Hybridisation and Genome Chimerisation in Saccharomyces: Combining of Gene Pools of Species and Its Biotechnological Perspectives.

Over the last one and a half decade, interspecies hybridisation has gained continuously increasing attention as a breeding technique suitable for transferring of genetic information between Saccharomyces species and mixing of their gene pools without genetic engineering. The hybrids frequently show positive transgressive phenotypes. Segregation of the hybrid genome results in mosaic (chimeric) strains that can outperform both the parents and the hybrids or exhibit novel positive phenotypic properties. Mitotic segregation can take place during the vegetative propagation of the sterile allodiploid hybrid cells. Meiotic segregation becomes possible after genome duplication (tetraploidisation) if it is followed by break-down of sterility. The allotetraploid cells are seemingly fertile because they form viable spores. But because of the autodiploidisation of the meiosis, sterile allodiploid spores are produced and thus the hybrid genome does not segregate (the second sterility barrier). However, malsegregation of MAT-carrying chromosomes in one of the subgenomes during allotetraploid meiosis (loss of MAT heterozygosity) results in fertile alloaneuploid spores. The breakdown of (the second) sterility barrier is followed by the loss of additional chromosomes in rapid succession and recombination between the subgenomes. The process (genome autoreduction in meiosis or GARMe) chimerises the genome and generates strains with chimeric (mosaic) genomes composed of various combinations of the genes of the parental strains. Since one of the subgenomes is preferentially reduced, the outcome is usually a strain having an (almost) complete genome from one parent and only a few genes or mosaics from the genome of the other parent. The fertility of the spores produced during GARMe provides possibilities also for introgressive backcrossing with one or the other parental strain, but genome chimerisation and gene transfer through series of backcrosses always with the same parent is likely to be less efficient than through meiotic or mitotic genome autoreduction. Hybridisation and the evolution of the hybrid genome (resizing and chimerisation) have been exploited in the improvement of industrial strains and applied to the breeding of new strains for specific purposes. Lists of successful projects are shown and certain major trends are discussed.

Efficiency of Embryo Rescue Technology in St. Augustinegrass Breeding

St. Augustinegrass [Stenotaphrum secundatum (Walt.) Kuntze] is a popular warm‐season turfgrass in Gulf Coast regions of the United States. Diploid St. Augustinegrass produces fertile seed, but triploids and higher polyploids are mostly sterile. Polyploid St. Augustinegrass cultivars have been selected for their resistance to southern chinch bugs, St. Augustine Decline, and drought tolerance but lack the cold hardiness and shade tolerance of diploids. However, because of sterility barriers, the polyploid germplasm had not been fully used as breeding material until 2009 when we reported the development of interploid hybrids using embryo rescue technology. The objective of the present study was to backcross these first‐generation interploid hybrids with diploid males to produce second‐generation hybrids. Sterility barriers were found to be much greater with second‐generation crosses yielding only 0.00 to 2.69% progeny per interploid hybrid. Using this technique, the embryos are generally rescued 3 wk postpollination, thereby causing a heavy demand on one's time to process spikelets before embryos abort. Our preliminary findings show the use of cold storage at 7°C to potentially extend the processing period up to 2 mo with a slight drop off in efficiency (4.37–6.03% crossability). Our results also show the potential of first‐generation hybrids to exhibit superior performance under drought when compared with diploid cultivars Palmetto and Raleigh. Tested second‐generation interploids did not exhibit superior performance in drought. Nonetheless, our results emphasize the utility of embryo rescue technology in facilitating gene exchange between ploidy levels to generate hybrids with superior performance.

Utilizing Wild Species for Peanut Improvement

The cultivated peanut (Arachis hypogaeaL.) is an allotetraploid species with a very large and complex genome. This species is susceptible to numerous foliar and soil‐borne diseases for which only moderate levels of resistance have been identified in the germplasm collection, but several of the 81 wild species are extremely resistant to many destructive peanut diseases. Peanut species were grouped into nine sections, but only taxa in sectionArachiswill hybridize withA. hypogaea. Most of these species are diploid, but two aneuploids and two tetraploids also exist in the section. The first peanut cultivars released after interspecific hybridization were ‘Spancross’ and ‘Tamnut 74’ during the 1970s from a cross betweenA. hypogaeaand its tetraploid progenitor. However, introgression of useful genes from diploids has been difficult due to sterility barriers resulting from genomic and ploidy differences. To utilize diploids in sectionArachis, direct hybrids have been made betweenA. hypogaeaand diploid species, the chromosome number doubled to the hexaploid level, and then tetraploids recovered with resistances to nematodes, leaf spots, rust, and numerous insect pests. ‘Bailey’, a widely grown Virginia‐type peanut, was released from these materials, and other cultivars are gown in Asia and South America. Alternatively, hybrids between diploid A and B genome species have been made, the chromosome number doubled, and cultivars released with nematode resistance derived fromArachisspecies. Introgression fromArachisspecies toA. hypogaeaappears to be in large blocks rather than as single genes, and new genotyping strategies should enhance utilization of wild peanut genetic resources.

Non-introgressive genome chimerisation by malsegregation in autodiploidised allotetraploids during meiosis of Saccharomyces kudriavzevii x Saccharomyces uvarum hybrids.

Saccharomyces strains with chimerical genomes consisting of mosaics of the genomes of different species ("natural hybrids") occur quite frequently among industrial and wine strains. The most widely endorsed hypothesis is that the mosaics are introgressions acquired via hybridisation and repeated backcrosses of the hybrids with one of the parental species. However, the interspecies hybrids are sterile, unable to mate with their parents. Here, we show by analysing synthetic Saccharomyces kudriavzevii x Saccharomyces uvarum hybrids that mosaic (chimeric) genomes can arise without introgressive backcrosses. These species are biologically separated by a double sterility barrier (sterility of allodiploids and F1 sterility of allotetraploids). F1 sterility is due to the diploidisation of the tetraploid meiosis resulting in MAT a /MAT α heterozygosity which suppresses mating in the spores. This barrier can occasionally be broken down by malsegregation of autosyndetically paired chromosomes carrying the MAT loci (loss of MAT heterozygosity). Subsequent malsegregation of additional autosyndetically paired chromosomes and occasional allosyndetic interactions chimerise the hybrid genome. Chromosomes are preferentially lost from the S. kudriavzevii subgenome. The uniparental transmission of the mitochondrial DNA to the hybrids indicates that nucleo-mitochondrial interactions might affect the direction of the genomic changes. We propose the name GARMe (Genome AutoReduction in Meiosis) for this process of genome reduction and chimerisation which involves no introgressive backcrossings. It opens a way to transfer genetic information between species and thus to get one step ahead after hybridisation in the production of yeast strains with beneficial combinations of properties of different species.

In situ morphometric survey elucidates the evolutionary systematics of the Eurasian Himantoglossum clade (Orchidaceae: Orchidinae).

Background and AimsThe charismatic Himantoglossum s.l. clade of Eurasian orchids contains an unusually large proportion of taxa that are of controversial circumscriptions and considerable conservation concern. Whereas our previously published study addressed the molecular phylogenetics and phylogeography of every named taxon within the clade, here we use detailed morphometric data obtained from the same populations to compare genotypes with associated phenotypes, in order to better explore taxonomic circumscription and character evolution within the clade.MethodsBetween one and 12 plants found in 25 populations that encompassed the entire distribution of the Himantoglossum s.l. clade were measured in situ for 51 morphological characters. Results for 45 of those characters were subjected to detailed multivariate and univariate analyses.Key ResultsMultivariate analyses readily separate subgenus Barlia and subgenus Comperia from subgenus Himantoglossum, and also the early-divergent H. formosum from the less divergent remainder of subgenus Himantoglossum. The sequence of divergence of these four lineages is confidently resolved. Our experimental approach to morphometric character analysis demonstrates clearly that phenotypic evolution within Himantoglossum is unusually multi-dimensional.ConclusionsDegrees of divergence between taxa shown by morphological analyses approximate those previously shown using molecular analyses. Himantoglossum s.l. is readily divisible into three subgenera. The three sections of subgenus Himantoglossum—hircinum, caprinum and formosum—are arrayed from west to east with only limited geographical overlap. At this taxonomic level, their juxtaposition combines with conflict between contrasting datasets to complicate attempts to distinguish between clinal variation and the discontinuities that by definition separate bona fide species. All taxa achieve allogamy via food deceit and have only weak pollinator specificity. Artificial crossing demonstrates that intrinsic sterility barriers are weak. Although we have found evidence of gene flow among and within the three sections of subgenus Himantoglossum, reports of natural hybrids are surprisingly rare, probably because putative parents are sufficiently similar to questionably warrant the status of species. Phenological separation and increased xeromorphy characterise the origin of subgenus Barlia. Several individual morphological characters show evidence of parallel acquisition, and loss of features is especially frequent in floral markings among members of section caprinum. Detailed patterns of gain and loss demonstrate that several different categories of flower markings are inherited independently. Along with the dimensions of labellar lobes, these pigmentation characters have been over-emphasised in previous taxonomic treatments. Increased plant vigour was a crucial element of the origin of the genus, but vegetative characters underwent remarkably little subsequent evolution. Attempts to reconstruct hypothetical ancestors at internal nodes of the phylogeny are weakened by (a) uncertain placement of Steveniella as sister to Himantoglossum s.l. and (b) uncertain relationships among subtly different putative species within section caprinum. Nonetheless, heterochronic/allometric trends, ultimately limited by functional constraints, clearly dictate transitions between contrasting flower sizes and complex labellum shapes.

Both morph- and species-dependent asymmetries affect reproductive barriers between heterostylous species.

The interaction between floral traits and reproductive isolation is crucial to explaining the extraordinary diversity of angiosperms. Heterostyly, a complex floral polymorphism that optimizes outcrossing, evolved repeatedly and has been shown to accelerate diversification in primroses, yet its potential influence on isolating mechanisms remains unexplored. Furthermore, the relative contribution of pre‐ versus postmating barriers to reproductive isolation is still debated. No experimental study has yet evaluated the possible effects of heterostyly on pre‐ and postmating reproductive mechanisms. We quantify multiple reproductive barriers between the heterostylous Primula elatior (oxlip) and P. vulgaris (primrose), which readily hybridize when co‐occurring, and test whether traits of heterostyly contribute to reproductive barriers in unique ways. We find that premating isolation is key for both species, while postmating isolation is considerable only for P. vulgaris; ecogeographic isolation is crucial for both species, while phenological, seed developmental, and hybrid sterility barriers are also important in P. vulgaris, implicating sympatrically higher gene flow into P. elatior. We document for the first time that, in addition to the aforementioned species‐dependent asymmetries, morph‐dependent asymmetries affect reproductive barriers between heterostylous species. Indeed, the interspecific decrease of reciprocity between high sexual organs of complementary floral morphs limits interspecific pollen transfer from anthers of short‐styled flowers to stigmas of long‐styled flowers, while higher reciprocity between low sexual organs favors introgression over isolation from anthers of long‐styled flowers to stigmas of short‐styled flowers. Finally, intramorph incompatibility persists across species boundaries, but is weakened in long‐styled flowers of P. elatior, opening a possible backdoor to gene flow through intramorph pollen transfer between species. Therefore, patterns of gene flow across species boundaries are likely affected by floral morph composition of adjacent populations. To summarize, our study highlights the general importance of premating isolation and newly illustrates that both morph‐ and species‐dependent asymmetries shape boundaries between heterostylous species.

Identifying Arachis Amphidiploids Resistant to Foliar Fungal Diseases

Many wild species of the genus Arachis are more resistant to pests and diseases than the cultivated species A. hypogaea. However, the use of Arachis germplasm in breeding programs is hampered by sterility barriers mainly due to differences between the genomes and the ploidy levels of A. hypogaea and related species. The aim of this study was to evaluate eight newly obtained amphidiploids and their parents for resistance to early leaf spot, late leaf spot, and rust under field conditions for three consecutive years. The diseases were evaluated using a diagrammatic scale with visual symptom scores ranging from 1 to 9 for the leaf spots and rust diseases. With few exceptions, all parental wild species accessions and the derived amphidiploids were more resistant than A. hypogaea checks. The amphidiploids with the greatest resistance to the three foliar fungal diseases were (A. magna Krapov., W.C. Gregory & C.E. Simpson V 13751 × A. cardenasii Krapov. & W.C. Gregory GKP 10017)4x, (A. magna K 30097 × A. stenosperma Krapov. & W.C. Gregory V 15076)4x, and (A. vallsii V 7635 × A. stenosperma V 10229)4x. These amphidiploids will be used in crosses and backcrosses with A. hypogaea, with the aim of performing introgression and pyramidization of disease resistance genes in cultivated peanut.

Two Tightly Linked Genes at the hsa1 Locus Cause Both F1 and F2 Hybrid Sterility in Rice

Molecular mechanisms of hybrid breakdown associated with sterility (F2 sterility) are poorly understood as compared with those of F1 hybrid sterility. Previously, we characterized three unlinked epistatic loci, hybrid sterility-a1 (hsa1), hsa2, and hsa3, responsible for the F2 sterility in a cross between Oryza sativa ssp. indica and japonica. In this study, we identified that the hsa1 locus contains two interacting genes, HSA1a and HSA1b, within a 30-kb region. HSA1a-j (japonica allele) encodes a highly conserved plant-specific domain of unknown function protein (DUF1618), whereas the indica allele (HSA1a-is) has two deletion mutations that cause disruption of domain structure. The second gene, HSA1b-is, encodes an uncharacterized protein with some similarity to a nucleotide-binding protein. Homozygous introgression of indica HSA1a-is–HSA1b-is alleles into japonica showed female gamete abortion at an early mitotic stage. The fact that the recombinant haplotype HSA1a-j–HSA1b-is caused semi-sterility in the heterozygous state with the HSA1a-is–HSA1b-is haplotype suggests that variation in the hsa1 locus is a possible cause of the wide-spectrum sterility barriers seen in F1 hybrids and successive generations in rice. We propose a simple genetic model to explain how a single causal mechanism can drive both F1 and F2 hybrid sterility.

Inheritance of Resistance to Downy Mildew in Sweet Basil

Sweet basil (Ocimum basilicum) is one of the most economically important culinary herbs in the world, yet global production has become increasingly challenging due to the destructive disease downy mildew (Peronospora belbahrii). Although multiple sources of resistance have been identified, there are no resistant sweet basil cultivars with a commercially acceptable chemotype and phenotype available. The commercial basil cultivar Mrihani (MRI) was identified as resistant and crossed with a Rutgers University susceptible sweet basil inbred line (SB22) to generate a full-sibling family. To determine the mode of inheritance for resistance to downy mildew in basil, six related generations of the MRI × SB22 family were evaluated using a disease severity index (DSI) at northern and southern New Jersey locations over 2 years. All siblings in the F1 and BC1P2 generations were resistant (0.33 > DSI) providing strong evidence that inheritance of resistance from MRI was conferred by dominant alleles. Segregation ratios in the F2 and backcross to the susceptible parent (BCP1) generations demonstrated chi-square goodness of fit to the two-gene complementary (F2: P = 0.11, BC1P1: P = 0.04) and recessive epistatic (F2: P = 0.03, BC1P1: P = 0.63) models. Further analyses of gene effects using a weighted six-parameter scaling test provided evidence that nonallelic additive × additive and additive × dominant gene effects were highly significant (P < 0.001) and resistance reducing. This is the first report of heritable genetic resistance that can be introduced to sweet basil without the issue of sterility barriers. Plant breeding strategies using the MRI × SB22 family should exploit dominant gene action and remove recessive, resistance-reducing alleles from the population.

English

Saffron (Crocus sativus L.) being triploid in nature is propagated by vegetative means through corms. The natural propagation rate of such plant species is relatively low, therefore some breeding and biotechnological technique like introduction, clonal selection, mutagenesis, micro-propagation and molecular markers have been used earlier as an alternative method of propagation for saffron. The creation of a germplasm bank consisting of superior elite clonal selections can be considered as a great achievement and in addition, the identification of selections as sources of variation can play an important role in improvement of this crop. The use of mutagenesis could increase the natural variability for important characters and may help in overcoming sterility barrier in autotriploid saffron by colchinization. Genetic improvement through mutation is one more important research area in saffron crop improvement and efforts made by different scientists have already given a base line idea for the improvement of this crop. Molecular markers have very good potential for study of species/clone level variation within saffron species. Use of molecular markers in saffron crop improvement will help in identifying the accessions and species which will be used for commercial exploitation and making of hybrids. At present we are having very few SSR markers in saffron, therefore both genomic and genic SSR markers need to be developed so that variation at genomic and expression level can be exploited.   Key words: Saffron, hybridization, clonal selection, introduction, mutagenesis, in-vitro regeneration, molecular markers.

Genetics of Cryptic Speciation within an Arctic Mustard, Draba nivalis

Crossing experiments indicate that hybrid sterility barriers frequently have developed within diploid, circumpolar plant species of the genus Draba. To gain insight into the rapid evolution of postzygotic reproductive isolation in this system, we augmented the linkage map of one of these species, D. nivalis, and searched for quantitative trait loci (QTLs) associated with reproductive isolation. The map adds 63 new dominant markers to a previously published dataset of 31 co-dominant microsatellites. These markers include 52 amplified fragment length polymorphisms (AFLPs) and 11 sequence-specific amplified polymorphisms (SSAPs) based on retrotransposon sequence. 22 markers displaying transmission ratio distortion were further included in the map. We resolved eight linkage groups with a total map length of 894 cM. Significant genotype-trait associations, or quantitative trait loci (QTL), were detected for reproductive phenotypes including pollen fertility (4 QTLs), seed set (3 QTLs), flowering time (3 QTLs) and number of flowers (4 QTLs). Observed patterns of inheritance were consistent with the influence of both nuclear-nuclear interactions and chromosomal changes on these traits. All seed set QTLs and one pollen fertility QTL displayed underdominant effects suggestive of the involvement of chromosomal rearrangements in hybrid sterility. Interestingly, D. nivalis is predominantly self-fertilizing, which may facilitate the establishment of underdominant loci and contribute to reproductive isolation.

Domestication and Long-Distance Dissemination of Rice: A Revised Version

Historically, in 1930s two sub-species, indica and japonica, were proposed on the basis of sterility in F1 hybrids between them. Subsequently, the two types were classified by associations of a number of genetically independent traits. The characteristic associations of traits have been explained by a series of hybrid sterility or reproductive barriers, in which a duplicate recessive gene model was proposed to cause varietal differentiation. In 1990s, most of hybrid sterility genes were proved to be caused by an interaction of allelic genes at each locus. In a well-known case of hybrid sterility, the indica, japonica, and wide-compatibility type which gives fertile hybrids in its cross to indica and japonica type, are proved to contain an allele, S5-i, S5-j and S5-n, respectively at a locus on Chromosome 6. Those gametes having S5-j allele is found to be partially aborted in the hybrid genotypes of S5-i/ S5-j while no gamete abortion occurred in S5-i/S5-n and S5-j/S5-n genotype. Since then, the S5-n has been used in hybrid rice breeding to obtain fertile and vigorous hybrids between indica and japonica types. For the last decade the action of S5-n is found to be due to a deletion in its gene sequence, while functional enzymes are produced by other alleles. Thus, the earlier assumed role of hybrid sterility genes as to lead to varietal differentiation are no more supported. Today, the characteristic association of traits found in each of varietal groups is better explained by founder effects. Recently , genetic analyses have been advanced for the contrasting traits between indica and japonica group. For an instance, genetic bases for long slender grain of indica and short wide grain of japonica are analyzed, and the short wide type is found to be caused by a deletion in its gene sequence. Thus, the short wide grain is considered to be mutational origin during domestication. On the other hand, a large number of native cultivars of rice were surveyed with enzyme polymorphism in 1980s, and later with molecular markers. As the results, profound genetic diversity is found in the genetic background of rice and wild rice. As the results, some scientist proposed the idea of multiple independent domestications of rice. However, before reaching to such a conclusion, at least two factors, i.e., long-distance-dissemination of some genotypes and possibilities of introgression by local wild rice to primitive cultivars need to be examined. Taking the two factors as well as historical events into consideration, the author considered that perennial japonica cultivars were first domesticated in Changjiang river basin, and disseminated to East India through Assam or along Bengal Bay, meanwhile their genetic background were transformed under introgressions of local wild rice which resulted to form the indica types. Thus–formed indica types seem to be disseminated to Southeast Asia, for an instance to Champa, under the expansion of Hinduism. Later some of the genotypes were introduced into China. Recently, this explanation has been verified by extensive analyses of gene sequence for wild and cultivated rice.

HYBRID INCOMPATIBILITY IS ACQUIRED FASTER IN ANNUAL THAN IN PERENNIAL SPECIES OF SUNFLOWER AND TARWEED

Hybrid sterility is an important species barrier, especially in plants where hybrids can often form between divergent taxa. Here we explore how life history affects the acquisition of hybrid sterility in two groups in the sunflower family. We analyzed genetic distance and F1 pollen sterility for interspecific crosses in annual and perennial groups. We find that reproductive isolation is acquired in a steady manner and that annual species acquire hybrid sterility barriers faster than perennial species. Potential causes of the observed sterility pattern are discussed.