A 2.09 Mb fragment translocation on chromosome 6 causes abnormalities during meiosis and leads to less seed watermelon

Seedlessness is a valuable agronomic trait in watermelon (Citrullus lanatus) breeding. Conventional less seed watermelons are mainly triploid, which has many disadvantages due to unbalanced genome content. Less seed watermelon can be achieved at the diploid level when certain reproductive genes are mutated or by chromosome translocation, which leads to defects during meiosis. However, the formation mechanism of diploid less seed watermelons remains largely unknown. Here, we identified a spontaneous mutant line, watermelon line “148”, which can set seeds normally when self-pollinated. A total of 148 × JM F1 hybrid plants exhibited seed number reductions to 50.3% and 47.3% of those of the two parental lines, respectively, which are considered to be less seed. Examination of pollen viability and hybridization experiments revealed that F1 hybrids produce semisterile pollen and ovules. Further cytological observations indicated that semisterility was a result of a reciprocal translocation of chromosomes, which exhibited one quadrivalent ring of four chromosomes at prometaphase I during meiosis. RT-qPCR analysis indirectly confirmed that the semisterile phenotype is caused by chromosome translocation rather than disruption of specific meiotic gene expression. F2 population genetic analysis indicated that the “148” watermelon line is a homozygous translocation and that the less seed phenotype of the F1 hybrid is prompted by one chromosome fragment translocation. The translocated fragment was further fine mapped to a 2.09 Mb region on chromosome 6 by whole-genome resequencing and genetic map cloning procedures. Our work revealed that a 2.09 Mb chromosome fragment translocation on chromosome 6, causing meiotic defects at metaphase I during meiosis, leads to diploid less seed watermelon. Our findings provide a new promising method for less seed watermelon breeding at the diploid level, as well as a fragment size reference for breeding less seed watermelon through artificially induced chromosome translocation.

Environmental and Genetic Factors Affecting Apospory Expressivity in Diploid Paspalum rufum

In angiosperms, gametophytic apomixis (clonal reproduction through seeds) is strongly associated with polyploidy and hybridization. The trait is facultative and its expressivity is highly variable between genotypes. Here, we used an F1 progeny derived from diploid apomictic (aposporic) genotypes of Paspalum rufum and two F2 families, derived from F1 hybrids with different apospory expressivity (%AES), to analyze the influence of the environment and the transgenerational transmission of the trait. In addition, AFLP markers were developed in the F1 population to identify genomic regions associated with the %AES. Cytoembryological analyses showed that the %AES was significantly influenced by different environments, but remained stable across the years. F1 and F2 progenies showed a wide range of %AES variation, but most hybrids were not significantly different from the parental genotypes. Maternal and paternal genetic linkage maps were built covering the ten expected linkage groups (LG). A single-marker analysis detected at least one region of 5.7 cM on LG3 that was significantly associated with apospory expressivity. Our results underline the importance of environmental influence in modulating apospory expressivity and identified a genomic region associated with apospory expressivity at the diploid level.

Targeted generation of polyploids in Hydrangea macrophylla through cross-based breeding

Background Up to now, diploid and triploid cultivars were reported for the ornamental crop Hydrangea macrophylla. Especially, the origin of triploids and their crossing behaviors are unknown, but the underlying mechanisms are highly relevant for breeding polyploids. Results By screening a cultivar collection, we identified diploid, triploid, tetraploid and even aneuploid H. macrophylla varieties. The pollen viability of triploids and tetraploids was comparable to that of diploids. Systematic crosses with these cultivars resulted in viable diploid, triploid, tetraploid and aneuploid offspring. Interestingly, crosses between diploids produced diploid and 0 or 1–94% triploid offspring, depending on the cultivars used as pollen parent. This finding suggests that specific diploids form unreduced pollen, either at low or high frequencies. In contrast, crosses of triploids with diploids or tetraploids produced many viable aneuploids, whose 2C DNA contents ranged between the parental 2C values. As expected, crosses between diploid and tetraploid individuals generated triploid offspring. Putative tetraploid plants were obtained at low frequencies in crosses between diploids and in interploid crosses of triploids with either diploid or tetraploid plants. The analysis of offspring populations indicated the production of 1n = 2x gametes for tetraploid plants, whereas triploids produced obviously reduced, aneuploid gametes with chromosome numbers ranging between haploid and diploid level. While euploid offspring grew normally, aneuploid plants showed mostly an abnormal development and a huge phenotypic variation within offspring populations, most likely due to the variation in chromosome numbers. Subsequent crosses with putative diploid, triploid and aneuploid offspring plants from interploid crosses resulted in viable offspring and germination rates ranging from 21 to 100%. Conclusions The existence of diploids that form unreduced pollen and of tetraploids allows the targeted breeding of polyploid H. macrophylla. Different ploidy levels can be addressed by combining the appropriate crossing partners. In contrast to artificial polyploidization, cross-based polyploidization is easy, cheap and results in genetically variable offspring that allows the direct selection of more robust and stress tolerant polyploid varieties. Furthermore, the generation of polyploid H. macrophylla plants will favor interspecific breeding programs within the genus Hydrangea.

Patterns of chromosomal variation in Mexican species of Aeschynomene (Fabaceae, Papilionoideae) and their evolutionary and taxonomic implications

A cytogenetic analysis of sixteen taxa of the genus Aeschynomene Linnaeus, 1753, which includes species belonging to both subgenera Aeschynomene (Léonard, 1954) and Ochopodium (Vogel, 1838) J. Léonard, 1954, was performed. All studied species had the same chromosome number (2n = 20) but exhibited karyotype diversity originating in different combinations of metacentric, submetacentric and subtelocentric chromosomes, chromosome size and number of SAT chromosomes. The plasticity of the genomes included the observation in a taxon belonging to the subgenus Aeschynomene of an isolated spherical structure similar in appearance to the extra chromosomal circular DNA observed in other plant genera. By superimposing the karyotypes in a recent phylogenetic tree, a correspondence between morphology, phylogeny and cytogenetic characteristics of the taxa included in the subgenus Aeschynomene is observed. Unlike subgenus Aeschynomene, the species of Ochopodium exhibit notable karyotype heterogeneity. However the limited cytogenetic information recorded prevents us from supporting the proposal of their taxonomic separation and raise it to the genus category. It is shown that karyotype information is useful in the taxonomic delimitation of Aeschynomene and that the diversity in the diploid level preceded the hybridization/polyploidization demonstrated in the genus. The systematic implications of our results and their value can be extended to other Dalbergieae genera as knowledge about the chromosomal structure and its evolution increases.

Production of hybrid potatoes: Are heterozygosity and ploidy levels important?

It has been proposed that maximizing heterosis for yield in potato may be achieved by maximizing heterozygosity and associated intra and interlocus interactions. Tetraploids offer more opportunities to create such interactions than diploids hence the general observations that tetraploids are higher yielding than diploids. Consequently, efforts have been made to increase heterozygosity in tetraploids by introgressing allelic diversity from other Solanum species into cultivated potato. However, conventional potato breeding is difficult because the cultivated potato is an autotetraploid with tetrasomic inheritance and it comprises highly heterozygous individuals which suffer inbreeding depression upon selfing; breeding at the tetraploid level is slow and less efficient than at diploid level. At the diploid level, it is possible to breed for and fix traits under recessive genetic control; it is nearly impossible to do so at the tetraploid level. There is also rapid response to selection due to greater variation in diploids than tetraploids. Consequently, there have been efforts to convert potato from an asexually propagated tetraploid crop into an inbred seed-propagated diploid; this is by production of inbred lines through selfing of the tetraploids to assemble desirable combinations of genes in the inbreds. These efforts are at the experimental stages and a lot of research needs to be done before they are confirmed. Because currently there is little experimental evidence to support superiority of the inbred seed-propagated diploid strategy, it appears the theory that heterosis for yield in potato may be achieved by maximizing heterozygosity and associated intra and interlocus interactions remain unchallenged; these interactions are more in tetraplods than in diploids. This paper therefore looks at genetic basis of yield heterosis in cultivated potato and the role of heterozygosity and ploidy level in production of hybrid potatoes.

SSR Based Genetic Diversity Analysis in Diploid Algaroba (Prosopis spp.) Population

Algaroba (Prosopis palida and Prosopis juliflora species) provides important environmental and economic benefits for semi-arid regions of the world. These are resistant to drought, and its fruits are used in the manufacture of flour and algarobina syrup. In the present study, the chromosome number, the ploidy level, and the genetic diversity based on 40 microsatellite loci of Prosopis spp. were determined in samples of a Brazilian algaroba population. The cytogenetic analysis in the metaphase showed only diploid individuals (2n = 28), with multiple cells featuring two CMA3/DAPI- heterochromatic blocks suggesting diploid level. However, polysomatism was found by the presence of some cells with four CMA3/DAPI- blocks, showing the tetraploid level just in some somatic cells. Among all of the primers tested for cross-amplification in algaroba, 22 were selected to characterize the samples. Thirteen loci revealed allele polymorphisms in the population and are recommended for future population studies and genetic improvement. The mean values of the analyzes showed low genetic diversity (two alleles per locus and HE = 0.181), reflecting the history of the introduction of algaroba in this sampled locality, and suggesting the genetic bottleneck and probable events of founders, as well as the characteristics of the species of this genera. However, amplified loci indicated low inbreeding (allelic fixation index of -0.007), although heterozygosis was higher than expected by the Hardy-Weinberg equilibrium. Therefore, this algaroba population is formed by diploid individuals and adjusts to the tendency of low number of alleles per locus SSR commonly observed in different species of Prosopis.

Chromosome count, meiotic abnormalities and pollen sterility in Lahaul sweetvetch (Hedysarum astragaloides Benth. ex Baker, Fabaceae), an endemic and threatened species from India

Male meiotic studies were carried out on eight different accessions of Hedysarum astragaloides Benth. ex Baker (Fabaceae), an endemic and threatened species of northwest Himalaya, India. Although genetic factors such as meiosis, chromosome number, and ploidy level may be causative for the evolution, endemism, rare distribution or even extinction of the species, no detailed information exists. Keeping this in mind H. astragaloides has been studied cytologically. Male meiotic investigations revealed diploid level (2n=2x=14) for species and normal meiotic course in the accessions from the Manali Hills resulting in nearly 100% pollen fertility. However, the accessions scored from the Manimahesh Hills and Pangi Valley depicted inter-pollen mother cell transfer of chromatin material and structural heterozygosity for reciprocal translocations. Consequent upon these meiotic anomalies, some pollen sterility (21%) resulted. On account of this sweeping genetic outcome, the incidence of anomalies such as this in an endemic and threatened species warrants grave consideration. It is sensible to conclude that conservation measures should include the collection of germplasm from the localities where plants are meiotically stable with high gametic fertility, to ensure good germination and healthy plants for future use. Seeds from meiotically normal individuals should be given priority for inclusion in seed banks.

Euchromatic Supernumerary Chromosomal Segments—Remnants of Ongoing Karyotype Restructuring in the Prospero autumnale Complex?

Supernumerary chromosomal segments (SCSs) represent additional chromosomal material that, unlike B chromosomes, is attached to the standard chromosome complement. The Prospero autumnale complex (Hyacinthaceae) is polymorphic for euchromatic large terminal SCSs located on the short arm of chromosome 1 in diploid cytotypes AA and B7B7, and tetraploid AAB7B7 and B6B6B7B7, in addition to on the short arm of chromosome 4 in polyploid B7B7B7B7 and B7B7B7B7B7B7 cytotypes. The genomic composition and evolutionary relationships among these SCSs have been assessed using fluorescence in situ hybridisation (FISH) with 5S and 35S ribosomal DNAs (rDNAs), satellite DNA PaB6, and a vertebrate-type telomeric repeat TTAGGG. Neither of the rDNA repeats were detected in SCSs, but most contained PaB6 and telomeric repeats, although these never spanned whole SCSs. Genomic in situ hybridisation (GISH) using A, B6, and B7 diploid genomic parental DNAs as probes revealed the consistently higher genomic affinity of SCSs in diploid hybrid B6B7 and allopolyploids AAB7B7 and B6B6B7B7 to genomic DNA of the B7 diploid cytotype. GISH results suggest a possible early origin of SCSs, especially that on chromosome 1, as by-products of the extensive genome restructuring within a putative ancestral P. autumnale B7 genome, predating the complex diversification at the diploid level and perhaps linked to B-chromosome evolution.

Tolerance to Caspofungin in Candida albicans Is Associated with at Least Three Distinctive Mechanisms That Govern Expression of FKS Genes and Cell Wall Remodeling

ABSTRACT Expanding echinocandin use to prevent or treat invasive fungal infections has led to an increase in the number of breakthrough infections due to resistant Candida species. Although it is uncommon, echinocandin resistance is well documented for Candida albicans, which is among the most prevalent bloodstream organisms. A better understanding is needed to assess the cellular factors that promote tolerance and predispose infecting cells to clinical breakthrough. We previously showed that some mutants that were adapted to growth in the presence of toxic sorbose due to loss of one chromosome 5 (Ch5) also became more tolerant to caspofungin. We found here, following direct selection of mutants on caspofungin, that tolerance can be conferred by at least three mechanisms: (i) monosomy of Ch5, (ii) combined monosomy of the left arm and trisomy of the right arm of Ch5, and (iii) an aneuploidy-independent mechanism. Tolerant mutants possessed cell walls with elevated chitin and showed downregulation of genes involved in cell wall biosynthesis, namely, FKS, located outside Ch5, and CHT2, located on Ch5, irrespective of Ch5 ploidy. Also irrespective of Ch5 ploidy, the CNB1 and MID1 genes on Ch5, which are involved in the calcineurin signaling pathway, were expressed at the diploid level. Thus, multiple mechanisms can affect the relative expression of the aforementioned genes, controlling them in similar ways. Although breakthrough mutations in two specific regions of FKS1 have previously been associated with caspofungin resistance, we found mechanisms of caspofungin tolerance that are independent of FKS1 and thus represent an earlier event in resistance development.