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Interspecific hybridization between two different Brassicaceae species, namely Brassica rapa ssp. pekinensis (♀) (AA, 2n = 2x = 20) and genetically modified Brassica napus (♂) (AACC, 2n = 4x = 38), was performed to study the transmission of a herbicide resistance gene from a tetraploid to a diploid Brassica species. Initially, four different GM B. napus lines were used for hybridization with B. rapa via hand pollination. Among the F1 hybrids, the cross involving the B. rapa (♀) × GM B. napus (♂) TG#39 line exhibited the highest recorded crossability index of 14.7 ± 5.7. However, subsequent backcross progenies (BC1, BC2, and BC3) displayed notably lower crossability indices. The F1 plants displayed morphological characteristics more aligned with the male parent B. napus, with significant segregation observed in the BC1 generation upon backcrossing with the recurrent parent B. rapa. By the BC2 and BC3 generations, the progeny stabilized, manifesting traits from both parents to varying degrees. Cytogenetic analysis revealed a substantial reduction in chromosome numbers, particularly in backcrossing progenies. BC1 plants typically exhibited 21–25 chromosomes, while BC2 progenies showed 21–22 chromosomes, and by the BC3 generation, stability was achieved with an average of 20 chromosomes. SSR marker analysis confirmed the progressive reduction of C-genome regions, retaining minimal C-genome-specific bands throughout successive backcrossing. Despite the extensive elimination of C-genome-specific genomic regions, the glyphosate resistance gene from the male parent B. napus was introgressed into BC3 progenies, suggesting that the glyphosate resistance gene located and introgressed in A-chromosome/genome regions of the Brassica plants. [ABSTRACT FROM AUTHOR]

Key message: Embryo abortion at the heart-shaped stage is the main reason for the failure of interspecific hybridization of hydrangea, and salicylic acid plays a key role during embryo abortion. Difficulties in obtaining seeds from interspecific hybridization between Hydrangea macrophylla and H. arborescens had severely restricted the process of breeding new hydrangea varieties. To clarify the cause of reproductive barriers, an interspecific hybridization was made between H. macrophylla 'Endless Summer' (female parent) and H. arborescens 'Annabelle' (male parent). The results showed that both parents’ floral organs developed normally, 'Annabelle' had high pollen viability (84.83% at 8 h after incubation), and the pollen tube could enter into the ovule of 'Endless Summer' at 72 h after pollination. Therefore, the pre-fertilization barrier was not the main reason for the failure of interspecific hybridization. However, observation of the embryo development by paraffin sections showed that the embryo was aborted at the heart-shaped stage. In addition, salicylic acid (SA) content was significantly higher (fourfold, P < 0.01) at 21 days after pollination (DAP) as compared to that of 17 DAP, which means SA may be closely correlated with embryo development. A total of 957 metabolites were detected, among which 78 were significantly different. During the embryo abortion, phenylpropanoids and polyketides were significantly down-regulated, while organic oxygen compounds were significantly up-regulated. Further analysis indicated that the metabolic pathway was enriched in the shikimic acid biosynthesis pathway, which suggests that more SA was synthesized. Taken together, it can be reasonably speculated that SA plays a key role leading to embryo abortion underlying the interspecific hybridization between Hydrangea macrophylla and H. arborescens. The result is helpful to direct the breeding of hydrangea through distant hybridization. [ABSTRACT FROM AUTHOR]

Main conclusion: The leaf color asymmetry found in the reciprocal hybrids C. hystrix × C. sativus (HC) and C. sativus × C. hystrix (CH) could be influenced by the CsPPR gene (CsaV3_1G038250.1). Most angiosperm organelles are maternally inherited; thus, the reciprocal hybrids usually exhibit asymmetric phenotypes that are associated with the maternal parent. However, there are two sets of organelle genomes in the plant cytoplasm, and the mechanism of reciprocal differences are more complex and largely unknown, because the chloroplast genes are involved besides mitochondrial genes. Cucumis spp. contains the species, i.e., cucumber and melon, which chloroplasts and mitochondria are maternally inherited and paternally inherited, respectively, serving as good materials for the study of reciprocal differences. In this study, leaf color asymmetry was observed in the reciprocal hybrids (HC and CH) derived from C. sativus (2n = 14, CC) and C. hystrix (2n = 24, HH), where the leaves of HC were found to have reduced chlorophyll content, abnormal chloroplast structure and lower photosynthetic capacity. Transcriptomic analysis revealed that the chloroplast development-related genes were differentially expressed in leaf color asymmetry. Genetic analysis showed that leaf color asymmetry was caused by the maternal chloroplast genome. Comparative analysis of chloroplast genomes revealed that there was no mutation in the chloroplast genome during interspecific hybridization. Moreover, a PPR gene (CsaV3_1G038250.1) with RNA-editing function was found to be involved in the regulation of leaf color asymmetry. These findings provide new insights into the regulatory mechanisms of asymmetric phenotypes in plant reciprocal crosses. [ABSTRACT FROM AUTHOR]

In certain tetraploid species resulting from interspecific hybridization, one parent's subgenome is known to selectively undergo DNA loss. The molecular mechanisms behind this remain unclear. In our study, we compared the genomes of a standard diploid species with two allotetraploid species from the Xenopus genus, both possessing L (longer) and S (shorter) homoeologous subgenomes. We observed substantial gene losses and intergenic DNA deletions in both the S and L subgenomes of the tetraploid species. Gene losses were around 1,000 to 3,000 for L and 4,000 to 6,000 for S, with especially prominent losses in the S subgenome. Many of these losses likely occurred shortly after interspecific hybridization in both L/S subgenomes. We also deduced frequent large inversions in the S subgenome. Upon reassessing transposon dynamics using updated genome databases, we reaffirmed heightened DNA transposon activity during the hybridization, as previously reported. We next investigated whether S subgenome-biased DNA loss could be correlated with the activation of DNA transposons following hybridization. Notably, distinct patterns were observed in the dynamics of DNA transposons between the L and S subgenomes. Several DNA transposon subfamilies correlated positively with DNA deletions in the S subgenome and negatively in the L subgenome. Based on these results, we propose a model that, upon and after hybridization between two related diploid Xenopus species, the mixture of their genomes resulted in the derepression of DNA transposons, especially in the S subgenome, leading to selective DNA loss in the S subgenome. [ABSTRACT FROM AUTHOR]

Interspecific hybrid rootstocks have the potential to improve the productivity and resilience of pistachio orchards in response to environmental stresses. This study aimed to produce inter-specific hybrids between P. vera and P. integerrima (In) and compare their early growth with their parents under chilling temperatures. Controlled pollination using In pollen was conducted on five pistachio cultivars including 'Kaleh-Ghuchi', 'Khanjari-Ghermze', 'Akbari', 'Khanjari-Sefid', 'Ohadi' and a local variety. The fruit set in female parents through controlled pollination of interspecific crossings was significantly lower than natural open pollination, resulting in a high percentage of blank seeds. Only 'Khenjari-Ghermze' and 'Kalleh-Ghoochi' produced a sufficient number of seeds for further experiment. The germination and survival rate of hybrid seeds were lower than that of pistachio cultivars (P

Background. Hordeum bulbosum L. is the only wild species of the genus Hordeum, the gene pool of which is successfully used in introgressive hybridization to increase the genetic diversity of H. vulgare L. When creating introgression forms based on hybrids of H. vulgare with H. bulbosum, it is necessary to take into consideration the ploidy of the parent species, and their genetic features that determine the intensity of the process of H. bulbosum chromosomes elimination in embryogenesis. The purpose of our investigation was to study the features associated with the elimination of bulbous barley chromosomes in embryos obtained in various combinations of crosses involving the tetraploid hybrid H. bulbosum with H. vulgare for its effective use in obtaining introgression lines of H. vulgare. Material and methods. The analysis was performed on 9-15 days old embryos obtained in crosses of the tetraploid hybrid F2 (H. bulbosum A17 (4x) × H. vulgare ‘Borwina’ (4x)) (HbHbHvHv) with ‘Igri’ (2x) and ‘Borwina’ (2x) barley varieties, with H. bulbosum A17 (4x), as well as during its self-pollination. The chromosomal composition of embryos of different ages was analyzed on squashed embryo slides using genomic in situ hybridization (GISH). Results. Among the 11-15 days old embryos obtained from self-pollination of the hybrid, approximately half of the studied ones were mixoploids, while in other embryos the majority of cells contained only the genomic material of H. vulgare. The elimination process was very intensive in crosses of the hybrid with the barley varieties ‘Igri’ (2x) and ‘Borwina’ (2x), and by day 10-11 after pollination cells containing only the genomic material of H. vulgare predominated in all embryos, regardless of the direction of crossing and the variety used. Hybrid embryos with a stable chromosomal composition resulted from a cross of a tetraploid hybrid with H. bulbosum A17 (4x). Embryos with recombinant H. vulgare chromosomes carrying alien genetic material of bulbous barley were identified in all types of crosses. Conclusion. The most efficient way for the mass production of introgression lines of cultivated barley varieties based on the partially fertile hybrid F2 (H. bulbosum A17 (4x) × H. vulgare ‘Borwina’ (4x)), is the crossing with varieties of H. vulgare (2x). The progeny from such crosses will contain only cultivated barley plants, among which it is possible to identify forms with the introgression of H. bulbosum and use already the first generation from their self-pollination for selecting H. vulgare lines carrying the genetic material of bulbous barley in both homologs.

Polyploids are essential in plant evolution and species formation, providing a rich genetic reservoir and increasing species diversity. Complex polyploids with higher ploidy levels often have a dosage effect on the phenotype, which can be highly detrimental to gametes, making them rare. In this study, offspring plants resulting from an autoallotetraploid (RRRC) derived from the interspecific hybridization between allotetraploid Raphanobrassica (RRCC, 2n = 36) and diploid radish (RR, 2n = 18) were obtained. Fluorescence in situ hybridization (FISH) using C-genome-specific repeats as probes revealed two main genome configurations in these offspring plants: RRRCC (2n = 43, 44, 45) and RRRRCC (2n = 54, 55), showing more complex genome configurations and higher ploidy levels compared to the parental plants. These offspring plants exhibited extensive variation in phenotypic characteristics, including leaf type and flower type and color, as well as seed and pollen fertility. Analysis of chromosome behavior showed that homoeologous chromosome pairing events are widely observed at the diakinesis stage in the pollen mother cells (PMCs) of these allopolyploids, with a range of 58.73% to 78.33%. Moreover, the unreduced C subgenome at meiosis anaphase II in PMCs was observed, which provides compelling evidence for the formation of complex allopolyploid offspring. These complex allopolyploids serve as valuable genetic resources for further analysis and contribute to our understanding of the mechanisms underlying the formation of complex allopolyploids. [ABSTRACT FROM AUTHOR]

SUMMARY: Aegilops longissima and Ae. sharonensis, being classified into the Sitopsis section of genus Aegilops, are distinct species both taxonomically and ecologically. Nevertheless, earlier observations indicate that the two species are not reproductively isolated to full extent and can inter‐bred upon secondary contact. However, the genomic underpinnings of the morpho‐ecological differentiation between the two foci species remained unexplored. Here, we resequenced 31 representative accessions of the two species and conducted in‐depth comparative genomic analyses. We demonstrate recurrent and ongoing natural hybridizations between Ae. longissima and Ae. sharonensis, and depict features of genome composition of the resultant hybrids at both individual and population levels. We also delineate genomic regions and candidate genes potentially underpinning the differential morphological and edaphic adaptations of the two species. Intriguingly, a binary morphology was observed in the hybrids, suggesting existence of highly diverged genomic regions that remain uneroded by the admixtures. Together, our results provide new insights into the molding effects of interspecific hybridization on genome composition and mechanisms preventing merge of the two species. Significance Statement: We demonstrate recurrent and ongoing natural hybridizations between Aegilops longissima and Ae. sharonensis, and depict features of genome composition of the resultant hybrids at both individual and population levels. [ABSTRACT FROM AUTHOR]

Hardy geranium is a popular ornamental plant with a high market demand. As a result, there is a strong need for continuous innovation to improve its ornamental qualities. Interspecific hybridization is a widely used technique for introducing desirable traits and creating new cultivars with added value. To explore the possibilities of creating novelty within the Geranium genus, we conducted 2438 interspecific crosses over three flowering seasons, resulting in the successful harvest of 445 seeds and the final production of 82 seedlings. Using embryo rescue increased the germination rate of a seedling from 0.17% to 1.01%. To validate the paternal genetic contribution in the new seedlings, we used amplified fragment length polymorphism (AFLP) to assess the proportion of uniquely inherited paternal markers in all seedlings. AFLP analysis confirmed that 54 of the 82 seedlings were indeed true hybrids (65.8% hybridization success rate). A morphological evaluation of the flowers and leaves, including flower diameter (mm), style length (mm), number of flowers, flowering period (weeks), and color of both flowers and leaves, provided further confirmation of the F1 hybrid status of the seedlings. The results of our interspecific breeding demonstrated the efficiency of interspecific hybridization in hardy geraniums and its potential to create distinctive and novel cultivars for the ornamental market. [ABSTRACT FROM AUTHOR]

For peanut, the lack of stable cytological markers is a barrier to tracking specific chromosomes, elucidating the genetic relationships between genomes and identifying chromosomal variations. Chromosome mapping using single-copy oligonucleotide (oligo) probe libraries has unique advantages for identifying homologous chromosomes and chromosomal rearrangements. In this study, we developed two whole-chromosome single-copy oligo probe libraries, LS-7A and LS-8A, based on the reference genome sequences of chromosomes 7A and 8A of Arachis duranensis. Fluorescence in situ hybridization (FISH) analysis confirmed that the libraries could specifically paint chromosomes 7 and 8. In addition, sequential FISH and electronic localization of LS-7A and LS-8A in A. duranensis (AA) and A. ipaensis (BB) showed that chromosomes 7A and 8A contained translocations and inversions relative to chromosomes 7B and 8B. Analysis of the chromosomes of wild Arachis species using LS-8A confirmed that this library could accurately and effectively identify A genome species. Finally, LS-7A and LS-8A were used to paint the chromosomes of interspecific hybrids and their progenies, which verified the authenticity of the interspecific hybrids and identified a disomic addition line. This study provides a model for developing specific oligo probes to identify the structural variations of other chromosomes in Arachis and demonstrates the practical utility of LS-7A and LS-8A. [ABSTRACT FROM AUTHOR]

Numerous studies show that species identification of representatives of the genus Alosa using various genetic markers is often difficult and the search for more specific biomarkers is required. For the first time we analyzed polymorphism of COI gene fragment of mitochondrial DNA of two representatives of this genus (A. tanaica and A. kessleri), supplemented with new data on A. immaculata, from the waters of the Ponto-Caspian basin in comparative aspect with other representatives of the herring (Clupeoidea) genera Alosa, Clupea, Clupeonella, Sprattus, and Sardinops. The main result was the conclusion that within the genus Alosa, it is not possible to identify species using the marker used. On the one hand, specimens collected from morphologically distinct individuals and identified as different species have the same haplotypes. On the other hand, samples belonging to different species differ from each other by an insignificant number of nucleotide substitutions and do not form independent clades on the phylogram and haplotype network. This indicates the absence of genetic differentiation between the studied samples of herrings of genus Alosa into separate species and species groups when using DNA barcoding based on the COI gene. The reasons for such a phenomenon may be the following: (1) incorrect identification of species in catches, since shads (Alosidae) have high morphological flexibility and in many species, the main external morphological characters often overlap; (2) recent time of speciation by the standards of biological evolution for shads of genus Alosa; (3) difference in proportion of interspecific hybrids, which can vary significantly between populations of the same species. [ABSTRACT FROM AUTHOR]

Abstract Background Interspecific hybridisation is a powerful tool for increasing genetic diversity in plant breeding programmes. Hexaploid wheat (Triticum aestivum, 2n = 42) × barley (Hordeum vulgare, 2n = 14) intergeneric hybrids can contribute to the transfer of agronomically useful traits by creating chromosome addition or translocation lines as well as full hybrids. Information on the karyotype of hybrid progenies possessing various combinations of wheat and barley chromosomes is thus essential for the subsequent breeding steps. Since the standard technique of chromosome in situ hybridisation is labour-intensive and requires specific skills. a routine, cost-efficient, and technically less demanding approach is beneficial both for research and breeding. Results We developed a Multiplex Polymerase Chain Reaction (MPCR) method to identify individual wheat and barley chromosomes. Chromosome-specific primer pairs were designed based on the whole genome sequences of ‘Chinese Spring’ wheat and ‘Golden Promise’ barley as reference cultivars. A pool of potential primers was generated by applying a 20-nucleotide sliding window with consecutive one-nucleotide shifts on the reference genomes. After filtering for optimal primer properties and defined amplicon sizes to produce an ordered ladder-like pattern, the primer pool was manually curated and sorted into four MPCR primer sets for the wheat A, B, and D sub-genomes, and for the barley genome. The designed MPCR primer sets showed high chromosome specificity in silico for the genome sequences of all 18 wheat and barley cultivars tested. The MPCR primers proved experimentally also chromosome-specific for the reference cultivars as well as for 13 additional wheat and four barley genotypes. Analyses of 16 wheat × barley F1 hybrid plants demonstrated that the MPCR primer sets enable the fast and one-step detection of all wheat and barley chromosomes. Finally, the established genotyping system was fully corroborated with the standard genomic in situ hybridisation (GISH) technique. Conclusions Wheat and barley chromosome-specific MPCR offers a fast, labour-friendly, and versatile alternative to molecular cytogenetic detection of individual chromosomes. This method is also suitable for the high-throughput analysis of distinct (sub)genomes, and, in contrast to GISH, can be performed with any tissue type. The designed primer sets proved to be highly chromosome-specific over a wide range of wheat and barley genotypes as well as in wheat × barley hybrids. The described primer design strategy can be extended to many species with precise genome sequence information.

Artificial hybrids between cultivated Avena species and wild Avena macrostachya that possess genes for resistance to biotic and abiotic stresses can be important for oat breeding. For the first time, a comprehensive study of genomes of artificial fertile hybrids Avena sativa × Avena macrostachya and their parental species was carried out based on the chromosome FISH mapping of satellite DNA sequences (satDNAs) and also analysis of intragenomic polymorphism in the 18S–ITS1–5.8S rDNA region, using NGS data. Chromosome distribution patterns of marker satDNAs allowed us to identify all chromosomes in the studied karyotypes, determine their subgenomic affiliation, and detect several chromosome rearrangements. Based on the obtained cytogenomic data, we revealed differences between two A. macrostachya subgenomes and demonstrated that only one of them was inherited in the studied octoploid hybrids. Ribotype analyses showed that the second major ribotype of A. macrostachya was species-specific and was not represented in rDNA pools of the octoploids, which could be related to the allopolyploid origin of this species. Our results indicate that the use of marker satDNAs in cytogenomic studies can provide important data on genomic relationships within Avena allopolyploid species and hybrids, and also expand the potential for interspecific crosses for breeding. [ABSTRACT FROM AUTHOR]

Genome size variation is a crucial aspect of plant evolution, influenced by a complex interplay of factors. Repetitive elements, which are fundamental components of genomic architecture, often play a role in genome expansion by selectively amplifying specific repeat motifs. This study focuses on Amomum, a genus in the ginger family (Zingiberaceae), known for its 4.4-fold variation in genome size. Using a robust methodology involving PhyloNet reconstruction, RepeatExplorer clustering, and repeat similarity-based phylogenetic network construction, we investigated the repeatome composition, analyzed repeat dynamics, and identified potential hybridization events within the genus. Our analysis confirmed the presence of four major infrageneric clades (A-D) within Amomum, with clades A-C exclusively comprising diploid species (2n = 48) and clade D encompassing both diploid and tetraploid species (2n = 48 and 96). We observed an increase in the repeat content within the genus, ranging from 84% to 89%, compared to outgroup species with 75% of the repeatome. The SIRE lineage of the Ty1-Copia repeat superfamily was prevalent in most analyzed ingroup genomes. We identified significant difference in repeatome structure between the basal Amomum clades (A, B, C) and the most diverged clade D. Our investigation revealed evidence of ancient hybridization events within Amomum, coinciding with a substantial proliferation of multiple repeat groups. This finding supports the hypothesis that ancient hybridization is a driving force in the genomic evolution of Amomum. Furthermore, we contextualize our findings within the broader context of genome size variations and repeatome dynamics observed across major monocot lineages. This study enhances our understanding of evolutionary processes within monocots by highlighting the crucial roles of repetitive elements in shaping genome size and suggesting the mechanisms that drive these changes. [ABSTRACT FROM AUTHOR]

Interspecific hybrid materials obtained from crosses between cultivated sunflower and some accessions of wild species of the genus Helianthus were studied. Samples of the annual wild species Helianthus annuus (w.f.), H. argophyllus and the perennial hexaploid species H.resinosus were used as paternal parents. In the crosses, five CMS lines, developed at DАI-General Toshevo, were used as the maternal parents. The embryo rescue method was applied to obtain hybrid seeds from the cross 830A x GT-M-046 (H. resinosus Small (2n=102). By applying the hybridization scheme “cultivated sunflower x wild species”, a sufficient amount of seeds were obtained for further morphological, phenological, biochemical and phytopathological studies of the hybrid progenies. The content of oil and oleic acid in the seeds was determined. Interspecific forms characterized with highest content of oil and oleic acid in the seeds were obtained from crosses 4499A x GT-E- 166, 4499A x GT-E-122, 830A x GT-E- 171, 4499A x GT-E-153, 3607A x GT-E- 046, 217A x GT-E-122 and 217A x GT-E- 126. The obtained hybrid materials can be successfully used as an initial breeding material to enrich the genetic plasma of cultivated sunflower and create high oleic lines and hybrids. [ABSTRACT FROM AUTHOR]

Hybridization is one of the key processes shaping lineage diversification, particularly in regions that experienced strong climate oscillations. The alpine biome with its rich history of glacial-interglacial cycles and complex patterns of species distribution shifts offers an excellent system to investigate the impact of gene flow on population dynamics and speciation, important issues for evolutionary biology and biodiversity conservation. In this study, we combined genomic data (DArTseq), chloroplast markers, and morphology to examine phylogenetic relationships and the permeability of species boundaries and their evolutionary outcomes among the alpine extremophilic species of Puccinellia (Poaceae) in the Pamir Mountains, a part of the Mountains of Central Asia biodiversity hotspot. We determined the occurrence of interspecific hybrids between P. himalaica and P. pamirica, which demonstrated almost symmetric ancestry from their parental species and did not show signals of introgression. According to our integrative revision, the natural hybrids between P. himalaica and P. pamirica should be classified as Puccinellia ×vachanica (pro species). Using approximate Bayesian computation for population history inference, we uncovered that P. himalaica hybridized with P. pamirica independently in multiple localities over the Holocene. Hybrids inherited the fine-scale genetic structure from their parental species, which developed these patterns earlier, during the Late Pleistocene. Hybridization had different consequences for the involved parental lineages, likely playing an important role in a continuing decline of P. himalaica in the Pamir Mountains over the Holocene. Our results show that P. himalaica should be considered a critically endangered species in the Pamir Mountains and could also be retreating across its entire range of distribution in High Mountain Asia. Using a comparative phylogeographic framework, we revealed the risk of extinction of a cold-adapted alpine species in a global biodiversity hotspot. This study highlights that genomics could unravel diversity trends under climate change and provides valuable evidence for conservation management. [ABSTRACT FROM AUTHOR]

Late blight (Phytophthora infestans), characterized by small water-soaked areas that rapidly enlarge to form purple-brown, oily appearing blotches, is the most destructive fungal disease that continues to stymie worldwide tomato production. Severe outbreaks have been recorded in tomato-growing areas throughout the world in recent years, and the vast majority of commercial tomato cultivars are extremely vulnerable to the late blight infection. The aim of this study was to screen the wild tomato Solanum habrochaites accessions LA1223, LA1353, LA1718, LA1777, LA2156, LA2167, and LA2556 under natural disease pressure conditions to find a possible source of resistance to late blight and also characterized these accessions for various morphological characters. Three S. habrochaites accessions (LA1777, LA2167, and LA2556) were resistant and superior to the susceptible check. With a PDI of 22.29, LA 1777 was shown to have the highest resistance, and as such, it may be used as a donor for late blight resistant tomato pre-breeding programmes in the near future. The success of high crossability (percentage) of some hybrid combinations using LA1777 as a male parent, such as LA3846 × LA1777 (77.77), LA2377 × LA1777 (62.50), and LA3317 × LA1777 (60.00), is encouraging for the development of pre-breeding genetic stocks as well as for the discovery of novel alleles in wild tomato populations. The morphological characterization of seven wild tomato accessions mainly confirms the guidelines of the International Plant Genetic Resources Institute. These accessions and their interspecific crosses showed wide variability in phenotypic and reproductive characters for plant height, flower color, exserted stigma, leaf margin, flowering time, fruit set, and other economic traits that will facilitate breeders in the development of superior tomato cultivars and F1 hybrids resistant to biotic stresses, as well as differentiate these S. habrochaites accessions for their potential use in genetic enhancement of cultivated tomatoes. [ABSTRACT FROM AUTHOR]

To improve the ε-PL production in wild-type strains of Streptomyces. albulus, Streptomyces. noursei, Streptomyces. rochei and Streptomyces. yunnanensis, the interspecific hybridization based on protoplast fusion was first performed. Two-species hybridizations failed to obtain hybrids with significant increase in ε-PL production, but four-species hybridizations succeed in acquiring many high-yield hybrids. 16S rDNA homology alignment and RAPD confirmed that the hybrid HX17 was restructured by integrating gene fragments from S. albulus and S. rochei with S. noursei as the carrier. S. noursei HX17 was subsequently suffered from mutagenesis and genome shuffling combining with multiple antibiotic resistance, and a mutant S. noursei GX6 was obtained with ε-PL yield of 2.23 g/L in shake-flask fermentation. In fed-batch fermentation, the ε-PL production of GX6 reached 47.2 g/L, which was increased by 95.6% to 136.8% over the wild parents. Ribosomal genes associated with antibiotics were sequenced and majority of mutant strains had mutations at different sites, indicating that the increase of antibiotic resistance was strongly associated with them. This research proved that combining interspecific hybridization with multiple antibiotic resistance was as an effective approach to rapidly improve the ε-PL production in Streptomyces species. [ABSTRACT FROM AUTHOR]

Background: Interspecific hybridisation is a powerful tool for increasing genetic diversity in plant breeding programmes. Hexaploid wheat (Triticum aestivum, 2n = 42) × barley (Hordeum vulgare, 2n = 14) intergeneric hybrids can contribute to the transfer of agronomically useful traits by creating chromosome addition or translocation lines as well as full hybrids. Information on the karyotype of hybrid progenies possessing various combinations of wheat and barley chromosomes is thus essential for the subsequent breeding steps. Since the standard technique of chromosome in situ hybridisation is labour-intensive and requires specific skills. a routine, cost-efficient, and technically less demanding approach is beneficial both for research and breeding. Results: We developed a Multiplex Polymerase Chain Reaction (MPCR) method to identify individual wheat and barley chromosomes. Chromosome-specific primer pairs were designed based on the whole genome sequences of 'Chinese Spring' wheat and 'Golden Promise' barley as reference cultivars. A pool of potential primers was generated by applying a 20-nucleotide sliding window with consecutive one-nucleotide shifts on the reference genomes. After filtering for optimal primer properties and defined amplicon sizes to produce an ordered ladder-like pattern, the primer pool was manually curated and sorted into four MPCR primer sets for the wheat A, B, and D sub-genomes, and for the barley genome. The designed MPCR primer sets showed high chromosome specificity in silico for the genome sequences of all 18 wheat and barley cultivars tested. The MPCR primers proved experimentally also chromosome-specific for the reference cultivars as well as for 13 additional wheat and four barley genotypes. Analyses of 16 wheat × barley F1 hybrid plants demonstrated that the MPCR primer sets enable the fast and one-step detection of all wheat and barley chromosomes. Finally, the established genotyping system was fully corroborated with the standard genomic in situ hybridisation (GISH) technique. Conclusions: Wheat and barley chromosome-specific MPCR offers a fast, labour-friendly, and versatile alternative to molecular cytogenetic detection of individual chromosomes. This method is also suitable for the high-throughput analysis of distinct (sub)genomes, and, in contrast to GISH, can be performed with any tissue type. The designed primer sets proved to be highly chromosome-specific over a wide range of wheat and barley genotypes as well as in wheat × barley hybrids. The described primer design strategy can be extended to many species with precise genome sequence information. [ABSTRACT FROM AUTHOR]

This study was designed to elucidate the physiological responses of three Lotus forage accessions to alkaline stress, and the influence of inoculating with Pantoea eucalypti endophyte strain on alkaline stress mitigation. A diploid L. corniculatus (Lc) accession, L. tenuis (Lt), and the interspecific hybrid Lt × Lc obtained from these two parental lines were exposed to alkaline stress (pH 8.2).Both Lt and the Lt × Lc hybrid are alkaline‐tolerant compared to Lc, based on observations that dry mass was not reduced under stress, and there were no chlorosis symptoms on leaf blades. In all three Lotus accessions, Fe2+ concentration under stress decreased in aerial parts and simultaneously increased in roots. Inoculation with P. eucalypti considerably increased Fe2+ content in shoots of all three Lotus forage species under alkaline treatment.Photochemical efficiency of PSII was affected in Lc accession only when exposed to alkaline treatment. However, when cultivated under alkalinity with inoculation, plants recovered and had photosynthetic parameters equivalent to those in the control treatment.Together, the results highlight the importance of inoculation with P. eucalypti, which contributes significantly to mitigating alkaline stress. All results provide useful information for improving alkaline tolerance traits of Lotus forage species and their interspecific hybrids. [ABSTRACT FROM AUTHOR]

Cultivated eggplant (Solanum melongena L.) is produced in many countries with temperate and tropical climates and has great economic importance. In recent years, resistance to biotic and abiotic stress conditions in addition to increasing yield and quality has gained importance in plant breeding including eggplant breeding. Therefore, the wild relative S. incanum is an important parent in breeding studies as it provides resistance to some important biotic and abiotic stresses for eggplant. It is possible to obtain a fully fertile hybrid between the two species, as well as to establish F2 lines. However, it is a mystery whether there is gene introgression from wild ancestor in F2 individuals. The phenotypic distinguishing and elimination in selection of the nonhybrid plants by classical breeding methods requires a long time and intensive labour. For this reason, there is a need for solutions in breeding studies in which gene transfers from wild relatives can be detected in the early development stages of plants. In this study, it was aimed to determine whether the F2 individuals obtained from interspecific cross of an inbred line P45 belonging to S. melongena x S. incanum crosses, when they were at the seedling stage, were S. melongena, S. incanum or carry genes from both ancestors by using T1480 COSII marker. Among a total of 94 F2 eggplant individuals and two parent eggplant species, it was determined that 51 individuals carried DNA fragments from both parental eggplant species, while 22 individuals belonging to the S. incanum species and 23 individuals belonging to the S. melongena species were identified. The resulting segregation ratio was 1:2:1, which was in accordance with Mendelian genetics. The results clearly showed that T1480 could be a useful marker for distinguishing whether seedling stage F2 plants carry genes from S. melongena, S. incanum, or both parents. As a conclusion, T1480 COSII marker can be used in eggplant breeding studies and will enable the plant improvement to progress more reliably than classical approaches. [ABSTRACT FROM AUTHOR]

Reproductive barriers are the single largest impediment in breeding of Solanum aethiopicum (Shum and Gilo cultivars). Knowledge of these barriers is important for prudent germplasm utilization and genetic improvement of the species. This study investigated compatibility barriers between S. aethiopicum and its relatives. A randomized complete block design and a full diallel mating method were used to evaluate crossability and floral traits of six genotypes of four different species (S. aethiopicum, S. macrocarpon, S. anguivi and S. incanum) across two seasons. Parameters assessed were petal opening time, stigma receptivity, anther dehiscence, pollen quantity, pollen viability and crossing success. Most flowers for all parental genotypes were opened by 08:00 am. Mean number of open flowers was higher (12 ± 6) for S. aethiopicum Shum (N11) and lowest (n = 3 ± 1) for S. macrocarpon (E12; (p < 0.001), with other genotypes being intermediate. Stigma receptivity differed (p < 0.001) among genotypes and seasons and was highest for N11 at 3.31 ± 1.32 and lowest for S. incanum (In1) at 2.24 ± 1.27; other genotypes were intermediate. All genotypes possessed high pollen viability (µ > 80%) though species differed (p < 0.01). All genotypes exhibited self-compatibility to varying degrees with N11 showing the highest fruit success (67.9 ± 15.6%), seeds per fruit (82 ± 51), and F1 germination rate (79 ± 18%). Interspecific crosses using S. macrocarpon, S. anguivi and S. incanum as females showed poor or no fruit and seed. N11 (S. aethiopicum Shum) was the top performer as a female; crosses with S. macrocarpon (N11xE12) showed highest fruit success (65.9 ± 26.5), with S. anguivi (N11xA1) showing the highest seed set (n = 56 ± 34) and S. incanum (N11xIN1) showing the highest F1 germination (64 ± 19). The F1 intraspecific cross between S. aethiopicum Shum (N4) and S. aethiopicum Gilo (G4) did not germinate. Female parent functioning may be a key feature for the failure of interspecific crosses where S. aethiopicum is a donor as observed by the unilateral incompatibility. [ABSTRACT FROM AUTHOR]

Abstract Background Drought is one of the important abiotic stresses that can significantly reduce crop yields. In India, about 24% of Brassica juncea (Indian mustard) cultivation is taken up under rainfed conditions, leading to low yields due to moisture deficit stress. Hence, there is an urgent need to improve the productivity of mustard under drought conditions. In the present study, a set of 87 B. carinata-derived B. juncea introgression lines (ILs) was developed with the goal of creating drought-tolerant genotypes. Method The experiment followed the augmented randomized complete block design with four blocks and three checks. ILs were evaluated for seed yield and its contributing traits under both rainfed and irrigated conditions in three different environments created by manipulating locations and years. To identify novel genes and alleles imparting drought tolerance, Quantitative Trait Loci (QTL) analysis was carried out. Genotyping-by-Sequencing (GBS) approach was used to construct the linkage map. Results The linkage map consisted of 5,165 SNP markers distributed across 18 chromosomes and spanning a distance of 1,671.87 cM. On average, there was a 3.09 cM gap between adjoining markers. A total of 29 additive QTLs were identified for drought tolerance; among these, 17 (58.6% of total QTLs detected) were contributed by B. carinata (BC 4), suggesting a greater contribution of B. carinata towards improving drought tolerance in the ILs. Out of 17 QTLs, 11 (64.7%) were located on the B genome, indicating more introgression segments on the B genome of B. juncea. Eight QTL hotspots, containing two or more QTLs, governing seed yield contributing traits, water use efficiency, and drought tolerance under moisture deficit stress conditions were identified. Seventeen candidate genes related to biotic and abiotic stresses, viz., SOS2, SOS2 like, NPR1, FAE1-KCS, HOT5, DNAJA1, NIA1, BRI1, RF21, ycf2, WRKY33, PAL, SAMS2, orf147, MAPK3, WRR1 and SUS, were reported in the genomic regions of identified QTLs. Conclusions The significance of B. carinata in improving drought tolerance and WUE by introducing genomic segments in Indian mustard is well demonstrated. The findings also provide valuable insights into the genetic basis of drought tolerance in mustard and pave the way for the development of drought-tolerant varieties.

Interspecific hybridization between two different Brassicaceae species, namely Brassica rapa ssp. pekinensis (♀) (AA, 2n = 2x = 20) and genetically modified Brassica napus (♂) (AACC, 2n = 4x = 38), was performed to study the transmission of a herbicide resistance gene from a tetraploid to a diploid Brassica species. Initially, four different GM B. napus lines were used for hybridization with B. rapa via hand pollination. Among the F1 hybrids, the cross involving the B. rapa (♀) × GM B. napus (♂) TG#39 line exhibited the highest recorded crossability index of 14.7 ± 5.7. However, subsequent backcross progenies (BC1, BC2, and BC3) displayed notably lower crossability indices. The F1 plants displayed morphological characteristics more aligned with the male parent B. napus, with significant segregation observed in the BC1 generation upon backcrossing with the recurrent parent B. rapa. By the BC2 and BC3 generations, the progeny stabilized, manifesting traits from both parents to varying degrees. Cytogenetic analysis revealed a substantial reduction in chromosome numbers, particularly in backcrossing progenies. BC1 plants typically exhibited 21–25 chromosomes, while BC2 progenies showed 21–22 chromosomes, and by the BC3 generation, stability was achieved with an average of 20 chromosomes. SSR marker analysis confirmed the progressive reduction of C-genome regions, retaining minimal C-genome-specific bands throughout successive backcrossing. Despite the extensive elimination of C-genome-specific genomic regions, the glyphosate resistance gene from the male parent B. napus was introgressed into BC3 progenies, suggesting that the glyphosate resistance gene located and introgressed in A-chromosome/genome regions of the Brassica plants.

Incorporating the centromere-specific histone H3 protein CENH3 into the centromeric nucleosomes is indispensable for accurate centromere function and balanced chromosome segregation in most eukaryotes, including higher plants. In the cell nuclei of interspecific hybrids, divergent centromeric DNAs cohabit and lead the corresponding parental chromosomes through the mitotic and meiotic cell divisions. Depending on the transmission of the parental chromosomes carrying the CENH3-encoding genes, CENH3 proteins from one or both parents may be present in these hybrids. The incorporation of parental CENH3 proteins into the divergent centromeres and their role in the chromosome elimination process in interspecific hybrids is still poorly understood. Here, we produced wheat x barley F1 hybrids that carried different combinations of barley chromosomes with genes encoding for either one (αCENH3) or both barley CENH3 protein variants (α-- and βCENH3). We generated specific antibodies distinguishing between the wheat CENH3 proteins and barley αCENH3 and applied them together with FISH probes to detect the precise pattern of parental CENH3 deposition into the wheat and barley centromeric nucleosomes. Analysis of somatic and meiotic nuclei of the wheat x barley hybrids revealed the plasticity of the maternal (wheat) CENH3 proteins to become incorporated into the paternal (barley) centromeric nucleosomes. However, no evidence for paternal CENH3 plasticity was detected in this study. The significance of the unilateral centromere plasticity and possible patterns of CENH3 incorporation into centromeres in interspecific hybrids are discussed. [ABSTRACT FROM AUTHOR]

Key message: Imbalanced chromosomes and cell cycle arrest, along with down-regulated genes in DNA damage repair and sperm cell differentiation, caused pollen abortion in synthetic allodiploid Brassica juncea hybrids. Interspecific hybridization is considered to be a major pathway for species formation and evolution in angiosperms, but the occurrence of pollen abortion in the hybrids is common, prompting us to recheck male gamete development in allodiploid hybrids after the initial combination of different genomes. Here, we investigated the several key meiotic and mitotic events during pollen development using the newly synthesised allodiploid B. juncea hybrids (AB, 2n = 2× = 18) as a model system. Our results demonstrated the partial synapsis and pairing of non-homologous chromosomes concurrent with chaotic spindle assembly, affected chromosome assortment and distribution during meiosis, which finally caused difference in genetic constitution amongst the final tetrads. The mitotic cell cycle arrest during microspore development resulted in the production of anucleate pollen cells. Transcription analysis showed that sets of key genes regulating cyclin (CYCA1;2 and CYCA2;3), DNA damage repair (DMC1, NBS1 and MMD1), and ubiquitin–proteasome pathway (SINAT4 and UBC) were largely downregulated at the early pollen meiosis stages, and those genes involved in sperm cell differentiation (DUO1, PIRL1, PIRL9 and LBD27) and pollen wall synthesis (PME48, VGDH11 and COBL10) were mostly repressed at the late pollen mitosis stages in the synthetic allodiploid B. juncea hybrids (AB). In conclusion, this study elucidated the related mechanisms affecting pollen fertility during male gametophyte development at the cytological and transcriptomic levels in the synthetic allodiploid B. juncea hybrids. [ABSTRACT FROM AUTHOR]

Okra is one of the important vegetable crops suffering from yield losses due to various biotic stresses. Crop wild relatives are reservoir of genetic resistance to biotic stresses but their utilization in crop breeding is restricted by various pre- and post-zygotic crossing barriers. In our study, multiple stress tolerant lines 'IC90476-1' of Abelmoschus manihot var. tetraphyllus and (IC141055) of Abelmoschus moschatus were crossed reciprocally with commercially grown variety Pusa A-4 of A. esculentus. Subsequently, only direct cross (Abelmoschus esculentus as female parent) F1s, amphidiploidized (C1), selfed and backcrossed with A. esculentus variety Pusa A-4 were raised to investigate the crossability barriers. Abelmoschus manihot var. tetraphyllus was found to be bi-directionally crossable with A. esculentus and not hampered by any serious pre- and post-zygotic barriers. But differences in ploidy leads to failing in production of F1 seeds upon selfing and backcrossing. Therefore, amphidiploidization of F1 hybrid followed by selfing and backcrossing (amphidiploid as seed parent) is recommended as an effective strategy to transfer desirable traits from A. manihot var. tetraphyllus to A. esculentus. However, A. moschatus × A. esculentus crosses suffered from severe pre-zygotic barriers that resulted in formation of aborted embryo with partially filled seeds. Therefore, use of A. esculentus only as female parent, in crossing with A. moschatus may increase the production of viable F1 seeds. In conclusion, colchicine (0.1%) induced amphidiploidization was found effective for restoration of fertility. However greater success was obtained in reciprocal backcrossing (A. esculentus × F1/C1) compared to amphidiploidization followed by selfing of C1 was recommended for gene introgression from A. moschatus. [ABSTRACT FROM AUTHOR]

The results of the ampelographic description of the newly selected Miro table grapes variety have been presented. It is an early ripening white table grapes variety, obtained by the method of interspecific hybridization at the Institute of Viticulture and Enology – Pleven. It was approved by the Executive Agency for Variety Testing, Field Inspection and Seed Control in 2019 and patented in 2023. The leaf blade is medium-sized, rounded, thick, lightly cut. The upper notches are shallow, open, in the form of a slit. The lower notches are shallow, in the form of a sharp angle, often barely noticeable. The petiole notch is most often closed, narrow with a pointed bottom, sometimes open with a lyreshaped opening. The cluster is very large, cylindrical-conical, semi-compact to loose. The berry is very large, oval, greenish yellow in colour. The variety is characterized by high fertility and yield and is distinguished by enhanced resistance to fungal diseases and low winter temperatures. The grapes have harmonious taste, very good transportability. They are suitable both for fresh consumption and after storage. The variety can be successfully grown in all wine-growing regions of the country and is appropriate for the production of bio table grapes. [ABSTRACT FROM AUTHOR]

Background: Drought is one of the important abiotic stresses that can significantly reduce crop yields. In India, about 24% of Brassica juncea (Indian mustard) cultivation is taken up under rainfed conditions, leading to low yields due to moisture deficit stress. Hence, there is an urgent need to improve the productivity of mustard under drought conditions. In the present study, a set of 87 B. carinata-derived B. juncea introgression lines (ILs) was developed with the goal of creating drought-tolerant genotypes. Method: The experiment followed the augmented randomized complete block design with four blocks and three checks. ILs were evaluated for seed yield and its contributing traits under both rainfed and irrigated conditions in three different environments created by manipulating locations and years. To identify novel genes and alleles imparting drought tolerance, Quantitative Trait Loci (QTL) analysis was carried out. Genotyping-by-Sequencing (GBS) approach was used to construct the linkage map. Results: The linkage map consisted of 5,165 SNP markers distributed across 18 chromosomes and spanning a distance of 1,671.87 cM. On average, there was a 3.09 cM gap between adjoining markers. A total of 29 additive QTLs were identified for drought tolerance; among these, 17 (58.6% of total QTLs detected) were contributed by B. carinata (BC 4), suggesting a greater contribution of B. carinata towards improving drought tolerance in the ILs. Out of 17 QTLs, 11 (64.7%) were located on the B genome, indicating more introgression segments on the B genome of B. juncea. Eight QTL hotspots, containing two or more QTLs, governing seed yield contributing traits, water use efficiency, and drought tolerance under moisture deficit stress conditions were identified. Seventeen candidate genes related to biotic and abiotic stresses, viz., SOS2, SOS2 like, NPR1, FAE1-KCS, HOT5, DNAJA1, NIA1, BRI1, RF21, ycf2, WRKY33, PAL, SAMS2, orf147, MAPK3, WRR1 and SUS, were reported in the genomic regions of identified QTLs. Conclusions: The significance of B. carinata in improving drought tolerance and WUE by introducing genomic segments in Indian mustard is well demonstrated. The findings also provide valuable insights into the genetic basis of drought tolerance in mustard and pave the way for the development of drought-tolerant varieties. [ABSTRACT FROM AUTHOR]

Cotton (Gossypium spp.) is the most important natural fiber source in the world. The genetic potential of cotton can be successfully and efficiently exploited by identifying and solving the complex fundamental problems of systematics, evolution, and phylogeny, based on interspecific hybridization of cotton. This study describes the results of interspecific hybridization of G. herbaceum L. (A1-genome) and G. mustelinum Miers ex Watt (AD4-genome) species, obtaining fertile hybrids through synthetic polyploidization of otherwise sterile triploid forms with colchicine (C22H25NO6) treatment. The fertile F1C hybrids were produced from five different cross combinations: (1) G. herbaceum subsp. frutescens × G. mustelinum; (2) G. herbaceum subsp. pseudoarboreum × G. mustelinum; (3) G. herbaceum subsp. pseudoarboreum f. harga × G. mustelinum; (4) G. herbaceum subsp. africanum × G. mustelinum; (5) G. herbaceum subsp. euherbaceum (variety A-833) × G. mustelinum. Cytogenetic analysis discovered normal conjugation of bivalent chromosomes in addition to univalent, open, and closed ring-shaped quadrivalent chromosomes at the stage of metaphase I in the F1C and F2C hybrids. The setting of hybrid bolls obtained as a result of these crosses ranged from 13.8–92.2%, the fertility of seeds in hybrid bolls from 9.7–16.3%, and the pollen viability rates from 36.6–63.8%. Two transgressive plants with long fiber of 35.1–37.0 mm and one plant with extra-long fiber of 39.1–41.0 mm were identified in the F2C progeny of G. herbaceum subsp. frutescens × G. mustelinum cross. Phylogenetic analysis with 72 SSR markers that detect genomic changes showed that tetraploid hybrids derived from the G. herbaceum × G. mustelinum were closer to the species G. mustelinum. The G. herbaceum subsp. frutescens was closer to the cultivated form, and its subsp. africanum was closer to the wild form. New knowledge of the interspecific hybridization and synthetic polyploidization was developed for understanding the genetic mechanisms of the evolution of tetraploid cotton during speciation. The synthetic polyploids of cotton obtained in this study would provide beneficial genes for developing new cotton varieties of the G. hirsutum species, with high-quality cotton fiber and strong tolerance to biotic or abiotic stress. In particular, the introduction of these polyploids to conventional and molecular breeding can serve as a bridge of transferring valuable genes related to high-quality fiber and stress tolerance from different cotton species to the new cultivars. [ABSTRACT FROM AUTHOR]

• Significant morphological diversity exists amongst the species- C. annuum, C. frutescens, C. chinense for all the characters studied. • Interspecific hybridization in Capsicum species is possible despite hindered by pre- and post-fertilization barriers. • Amongst the successful crosses made in the present study, Mem x King chilli-A5 had the highest fruit set (60 %), while C. annuum (Capsicum) x C. chinense (King chilli-A18) and King chilli-A18 x Krishna had the lowest fruit set (20 %). • It was found Capsicum x King chilli-A18(19.10 %) had the lowest pollen sterility and was the most desirable hybrid in terms of pollen fertility. • Estimation of the Usual Euclidean distance amongst the parents and their F 1 s revealed that Mem and F 1 of C. chinense (King chilli-A5) x C. frutescens (Mem) had the minimum distinction (94.39) amongst them, supporting that King chilli is the natural hybrid of C. chinense and C. frutescens. The present study explores the potentiality of interspecific crosses using three different Capsicum species, comprising of two C. annuum (Krishna and Capsicum) , five C. chinense (King chilli A1, A5, A18, A19, A20) and one C. frutescens (Mem) genotypes, with the objective to identify the incompatibility barriers, accomplish the morpho-molecular characterization of parents and hybrids and assess the genetic relatedness between the genotypes. Amongst the three species used for the crossing programme, 'Krishna' (C. annuum) and 'Mem' (C. frutescens) are local cultivars of Assam (India) and possess excellent adaptability. Guinness World Records certified King chilli, or 'Bhut Jolokia' (C. chinense), as the world's hottest chilli (10,001,304 SHU) in September 2007. King chilli has tremendous export potential but is prone to viral diseases, thereby necessitating the transfer of desirable disease resistance from local cultivars. It was observed from the study that the reciprocal crosses involving C. chinense and C. frutescens were compatible, but C. frutescens (Mem) was crossable with only accession A5, suggesting genotype specificity in the interspecific crosses. The usual Euclidean distance between Mem and A5 was 716.605 Similarly reciprocal crosses involving C. annuum and C. chinense were partially incompatible. Both the C. annuum (Krishna, Capsicum) genotypes successfully crossed with King chilli accession A18 thereby exhibiting genotype specificity. It was observed that in the cross A18 x Krishna the F 1 seeds failed to germinate and in the cross A18 x Capsicum the F 1 plants exhibited dwarfism. Hence, unilateral incompatibility was observed when King chilli accession A18 was used as the female parent. In the reciprocal crosses, germination and plant development was normal. The pollen viability ranged from 72.64 to 94.92 % for parents and 23.55 to 81.02 % for the hybrids. Thus, hybrids exhibited lower pollen viability compared to their parents. Significant morphological differences were visible between the parents and their combinations; some features were similar to the male or female parent, and others were either intermediate or novel. Parental polymorphism was detected using thirty random SSR primers, and the markers showing polymorphic bands were used to confirm the hybridity of F 1 s. Therefore, an attempt was made in this study to assess the possibility of interspecific crosses to transfer disease resistance from the related species to enhance the quality of King chilli, thus improving its commercial valve. [ABSTRACT FROM AUTHOR]

Cotton (Gossypium species) has received considerable interest from the geneticists, cytologists and evolutionary biologists since the last more than a century. Here, we explore the genetics of petal spot in the interspecific derivatives involving tetraploid and diploid cottons; and confirm the location of gene governing petal spot phenotype on chromosome A7 by demonstrating co-segregation of SSR marker NAU 2186 with petal spot phenotype. The presence of petal spot was observed to be dominant over its absence. Petal spot inheritance showed significant deviation from the expected Mendelian ratio in all the segregating populations indicating segregation distortion. The distortion was biased towards the hirsutum parent which has important implications from introgression point of view. We also report a strong association between petal spot and petal margin coloration phenotypes. Extant American cotton varieties generally lack petal spot and margin coloration phenotypes. These petal characteristics can serve as morphological markers during germplasm characterization.

Spelt (Triticum aestivum ssp. spelta) is an important wheat subspecies mainly cultivated in Europe before the 20th century that has contributed to modern wheat breeding as a valuable genetic resource. However, relatively little is known about the origins and maintenance of spelt populations. Here, using resequencing data from 416 worldwide wheat accessions, including representative spelt wheat, we demonstrate that European spelt emerged when primitive hexaploid wheat spread to the west and hybridized with pre-settled domesticated emmer, the putative maternal donor. Genomic introgression regions from domesticated emmer confer spelt’s primitive morphological characters used for species taxonomy, such as tenacious glumes and later flowering. We propose a haplotype-based “spelt index” to identify spelt-type wheat varieties and to quantify utilization of the spelt gene pool in modern wheat cultivars. This study reveals the genetic basis for the establishment of the spelt wheat subspecies in a specific ecological niche and the vital role of the spelt gene pool as a unique germplasm resource in modern wheat breeding.

Genome size variation is a crucial aspect of plant evolution, influenced by a complex interplay of factors. Repetitive elements, which are fundamental components of genomic architecture, often play a role in genome expansion by selectively amplifying specific repeat motifs. This study focuses on Amomum, a genus in the ginger family (Zingiberaceae), known for its 4.4-fold variation in genome size. Using a robust methodology involving PhyloNet reconstruction, RepeatExplorer clustering, and repeat similarity-based phylogenetic network construction, we investigated the repeatome composition, analyzed repeat dynamics, and identified potential hybridization events within the genus. Our analysis confirmed the presence of four major infrageneric clades (A–D) within Amomum, with clades A–C exclusively comprising diploid species (2n = 48) and clade D encompassing both diploid and tetraploid species (2n = 48 and 96). We observed an increase in the repeat content within the genus, ranging from 84% to 89%, compared to outgroup species with 75% of the repeatome. The SIRE lineage of the Ty1-Copia repeat superfamily was prevalent in most analyzed ingroup genomes. We identified significant difference in repeatome structure between the basal Amomum clades (A, B, C) and the most diverged clade D. Our investigation revealed evidence of ancient hybridization events within Amomum, coinciding with a substantial proliferation of multiple repeat groups. This finding supports the hypothesis that ancient hybridization is a driving force in the genomic evolution of Amomum. Furthermore, we contextualize our findings within the broader context of genome size variations and repeatome dynamics observed across major monocot lineages. This study enhances our understanding of evolutionary processes within monocots by highlighting the crucial roles of repetitive elements in shaping genome size and suggesting the mechanisms that drive these changes.

Hybridization is one of the key processes shaping lineage diversification, particularly in regions that experienced strong climate oscillations. The alpine biome with its rich history of glacial-interglacial cycles and complex patterns of species distribution shifts offers an excellent system to investigate the impact of gene flow on population dynamics and speciation, important issues for evolutionary biology and biodiversity conservation. In this study, we combined genomic data (DArTseq), chloroplast markers, and morphology to examine phylogenetic relationships and the permeability of species boundaries and their evolutionary outcomes among the alpine extremophilic species of Puccinellia (Poaceae) in the Pamir Mountains, a part of the Mountains of Central Asia biodiversity hotspot. We determined the occurrence of interspecific hybrids between P. himalaica and P. pamirica, which demonstrated almost symmetric ancestry from their parental species and did not show signals of introgression. According to our integrative revision, the natural hybrids between P. himalaica and P. pamirica should be classified as Puccinellia ×vachanica (pro species). Using approximate Bayesian computation for population history inference, we uncovered that P. himalaica hybridized with P. pamirica independently in multiple localities over the Holocene. Hybrids inherited the fine-scale genetic structure from their parental species, which developed these patterns earlier, during the Late Pleistocene. Hybridization had different consequences for the involved parental lineages, likely playing an important role in a continuing decline of P. himalaica in the Pamir Mountains over the Holocene. Our results show that P. himalaica should be considered a critically endangered species in the Pamir Mountains and could also be retreating across its entire range of distribution in High Mountain Asia. Using a comparative phylogeographic framework, we revealed the risk of extinction of a cold-adapted alpine species in a global biodiversity hotspot. This study highlights that genomics could unravel diversity trends under climate change and provides valuable evidence for conservation management.

Background. The study of the zone contacts of species, namely hybrid zones and sympatry zones, is an urgent task of evolutionary biology and modern zoology. Long-term monitoring of changes in the spatial and genetic structure of sympatry zones with parallel study of morphological, phenotypic and bioacoustic features of the hybrid population makes it possible to find out the mechanisms and consequences of the process of random interspecific hybridization. The purpose of the work is to monitor the state of the sympatry zone of Speckled and Russet Ground squirrels in the Volga region at the points of contact of the species and to identify interspecific hybrids in contact colonies by a complex of molecular genetic markers, by color indicators and by parameters of acoustic signals. Materials and methods. On the territory of Ulyanovsk region 9 colonies were surveyed: 6 combined colonies with specific separation on microstations, 3 mixed colonies without separation. In contact colonies, it was captured and marked 24 individuals of both species and possible interspecific hybrids. Molecular genetic analysis was conducted on 5 markers (D-loop, Hoxb5, р53, ZfX, SmcY) for determination of individuals’ origin. Sound signals and color indicators of 11 large, 11 speckled ground squirrels and 5 presumed interspecific hybrids were analyzed. Statistical analysis of obtained data were perform in STATISTICA 10.0. Results. A study of the sympatry zone of the large and speckled ground squirrel on the territory of the Ulyanovsk region showed that as of 2023, only 5 (out of 10 in 2019) contact colonies remained. In the colony “Kargino” for the first time during the entire period of monitoring work, a female with a genotype indicating her origin as a result of direct crossing of a female S. major (m) and a male S. suslicus was captured. Intermediate inheritance of traits of parental species by hybrids of russet and speckled ground squirrels, which appeared as a result of random hybridization in the sympatry zone, is shown. Conclusions. By 2023, a depressive situation had developed in the sympatry zone of the russet and speckled ground squirrels, which led to the emergence of random interspecific hybridization at the points of contact of species. Monitoring studies of long-term contact colonies of this pair of sympatric species should be considered promising and potentially effective for the tasks of studying the hybridization of ground squirrels of the Volga region.

Polyploids are essential in plant evolution and species formation, providing a rich genetic reservoir and increasing species diversity. Complex polyploids with higher ploidy levels often have a dosage effect on the phenotype, which can be highly detrimental to gametes, making them rare. In this study, offspring plants resulting from an autoallotetraploid (RRRC) derived from the interspecific hybridization between allotetraploid Raphanobrassica (RRCC, 2n = 36) and diploid radish (RR, 2n = 18) were obtained. Fluorescence in situ hybridization (FISH) using C-genome-specific repeats as probes revealed two main genome configurations in these offspring plants: RRRCC (2n = 43, 44, 45) and RRRRCC (2n = 54, 55), showing more complex genome configurations and higher ploidy levels compared to the parental plants. These offspring plants exhibited extensive variation in phenotypic characteristics, including leaf type and flower type and color, as well as seed and pollen fertility. Analysis of chromosome behavior showed that homoeologous chromosome pairing events are widely observed at the diakinesis stage in the pollen mother cells (PMCs) of these allopolyploids, with a range of 58.73% to 78.33%. Moreover, the unreduced C subgenome at meiosis anaphase II in PMCs was observed, which provides compelling evidence for the formation of complex allopolyploid offspring. These complex allopolyploids serve as valuable genetic resources for further analysis and contribute to our understanding of the mechanisms underlying the formation of complex allopolyploids.

Hardy geranium is a popular ornamental plant with a high market demand. As a result, there is a strong need for continuous innovation to improve its ornamental qualities. Interspecific hybridization is a widely used technique for introducing desirable traits and creating new cultivars with added value. To explore the possibilities of creating novelty within the Geranium genus, we conducted 2438 interspecific crosses over three flowering seasons, resulting in the successful harvest of 445 seeds and the final production of 82 seedlings. Using embryo rescue increased the germination rate of a seedling from 0.17% to 1.01%. To validate the paternal genetic contribution in the new seedlings, we used amplified fragment length polymorphism (AFLP) to assess the proportion of uniquely inherited paternal markers in all seedlings. AFLP analysis confirmed that 54 of the 82 seedlings were indeed true hybrids (65.8% hybridization success rate). A morphological evaluation of the flowers and leaves, including flower diameter (mm), style length (mm), number of flowers, flowering period (weeks), and color of both flowers and leaves, provided further confirmation of the F1 hybrid status of the seedlings. The results of our interspecific breeding demonstrated the efficiency of interspecific hybridization in hardy geraniums and its potential to create distinctive and novel cultivars for the ornamental market.

The formation process in the selection of winter common wheat for productivity and quality has been expanded by using the gene pool of related species. The purpose of this study was to expand the formative process in the selection of Triticum aestivum L. winter type of development with the further obtaining of genotypes with increased adaptability, plant productivity and grain quality through the use of winter common wheat with purple grain, whole grain wheat and Ethiopian wheat in hybridization. In the study, the methods of conducting the examination of plant varieties are used for carrying out phenological observations, morphological studies, evaluation of breeding material according to economic and valuable characteristics. In the creation of new forms, methods of hybridization and individual selection were used, and the TVEL method was used for pollination. The protein and gluten content were determined by infrared spectrometry. Parental components were studied and selected to expand the forming process to obtain productive grains from the main ear, thousand kernel weight, high grain quality, resistant to lodging, shedding of grain, drought-resistant and resistant to fungal pathogens, hybrid forms, specifically as a result of involvement of the source material Triticum aestivum L. and Triticum sphaerococcum Perc. in the hybridization. According to the results of the hybridization of winter common wheat and Ethiopian wheat, it was possible to obtain and breed in F4 forms with a long ear (>10-11 cm), an increased amount of grain from the main ear (>55 pcs.) and a different colour of grain from dark brown to purple, as an indicator of increased anthocyanin content and high antioxidant capacity. The practical value of this study lies in the fact that the research results expand information on the use of winter common wheat in the selection of productivity and quality of other types of wheat: Triticum sphaerococcum, T. aethiop. var. decoloratum, and the developed proposals will allow considering the use of parental components of wheat in research, help improve methods of hybridization and selection of new forms with desirable traits in the system of interspecies crossings

Incorporating the centromere-specific histone H3 protein CENH3 into the centromeric nucleosomes is indispensable for accurate centromere function and balanced chromosome segregation in most eukaryotes, including higher plants. In the cell nuclei of interspecific hybrids, divergent centromeric DNAs cohabit and lead the corresponding parental chromosomes through the mitotic and meiotic cell divisions. Depending on the transmission of the parental chromosomes carrying the CENH3-encoding genes, CENH3 proteins from one or both parents may be present in these hybrids. The incorporation of parental CENH3 proteins into the divergent centromeres and their role in the chromosome elimination process in interspecific hybrids is still poorly understood. Here, we produced wheat × barley F1 hybrids that carried different combinations of barley chromosomes with genes encoding for either one (αCENH3) or both barley CENH3 protein variants (α– and βCENH3). We generated specific antibodies distinguishing between the wheat CENH3 proteins and barley αCENH3 and applied them together with FISH probes to detect the precise pattern of parental CENH3 deposition into the wheat and barley centromeric nucleosomes. Analysis of somatic and meiotic nuclei of the wheat × barley hybrids revealed the plasticity of the maternal (wheat) CENH3 proteins to become incorporated into the paternal (barley) centromeric nucleosomes. However, no evidence for paternal CENH3 plasticity was detected in this study. The significance of the unilateral centromere plasticity and possible patterns of CENH3 incorporation into centromeres in interspecific hybrids are discussed.

The undomesticated rice relative Oryza longistaminata is a valuable genetic resource for the improvement of the domesticated Asian rice, Oryza sativa. To facilitate the conservation, management, and use of O. longistaminata germplasm, we sought to quantify the population structure and diversity of this species across its geographic range, which includes most of sub-Saharan Africa, and to determine phylogenetic relationships to other AA-genome species of rice present in Africa, including the prevalence of interspecific hybridization between O. longistaminata and O. sativa. Though past plant breeding efforts to introgress genes from O. longistaminata have improved biotic stress resistance, ratooning ability, and yield in O. sativa, progress has been limited by substantial breeding barriers. Nevertheless, despite the strong breeding barriers observed by plant breeders who have attempted this interspecific cross, there have been multiple reports of spontaneous hybrids of O. sativa and O. longistaminata (aka "Obake") obtained from natural populations in Africa. However, the frequency and extent of such natural introgressions and their effect on the evolution of O. longistaminata had not been previously investigated. We studied 190 O. longistaminata accessions, primarily from the International Rice Research Institute genebank collection, along with 309 O. sativa, 25 Oryza barthii, and 83 Oryza glaberrima control outgroups, and 17 control interspecific O. sativa/O. longistaminata hybrids. We analyzed the materials using 178,651 singlenucleotide polymorphisms (SNPs) and seven plastid microsatellite markers. This study identified three genetic subpopulations of O. longistaminata, which correspond geographically to Northwestern Africa, Pan-Africa, and Southern Africa. We confirmed that O. longistaminata is, perhaps counterintuitively, more closely related to the Asian species, O. sativa, than the African species O. barthii and O. glaberrima. We identified 19 recent spontaneous interspecific hybrid individuals between O. sativa and O. longistaminata in the germplasm sampled. Notably, the recent introgression between O. sativa and O. longistaminata has been bidirectional. Moreover, low levels of O. sativa alleles admixed in many predominantly O. longistaminata accessions suggest that introgression also occurred in the distant past, but only in Southern Africa. [ABSTRACT FROM AUTHOR]

Climate change is a major evolutionary force that can affect the structure of forest ecosystems worldwide. Red spruce (Picea rubens Sarg.) has recently faced a considerable decline in the Southern Appalachians due to rapid environmental change, which includes historical land use, and atmospheric pollution. In the northern part of its range, red spruce is sympatric with closely related black spruce (Picea mariana (Mill.) B.S.P.), where introgressive hybridization commonly occurs. We investigated range-wide population genetic diversity and structure and inferred postglacial migration patterns and evolution of red spruce using nuclear microsatellites. Moderate genetic diversity and differentiation were observed in red spruce. Genetic distance, maximum likelihood and Bayesian analyses identified two distinct population clusters: southern glacial populations, and the evolutionarily younger northern populations. Approximate Bayesian computation suggests that patterns of admixture are the result of divergence of red spruce and black spruce from a common ancestor and then introgressive hybridization during post-glacial migration. Genetic diversity, effective population size (Ne) and genetic differentiation were higher in the northern than in the southern populations. Our results along with previously available fossil data suggest that Picea rubens and Picea mariana occupied separate southern refugia during the last glaciation. After initial expansion in the early Holocene, these two species faced a period of recession and formed a secondary coastal refugium, where introgressive hybridization occurred, and then both species migrated northward. As a result, various levels of black spruce alleles are present in the sympatric red spruce populations. Allopatric populations of P. rubens and P. mariana have many species-specific alleles and much fewer alleles from common ancestry. The pure southern red spruce populations may become critically endangered under projected climate change conditions as their ecological niche may disappear. [ABSTRACT FROM AUTHOR]

Modern sugarcane cultivars (Saccharum spp., 2n = 100~120) are complex polyploids primarily derived from interspecific hybridization between S. officinarum and S. spontaneum. Nobilization is the theory of utilizing wild germplasm in sugarcane breeding, and is the foundation for utilizing S. spontaneum for stress resistance. However, the exact chromosomal transmission remains elusive due to a lack of chromosome-specific markers. Here, we applied chromosome-specific oligonucleotide (oligo)-based probes for identifying chromosomes 1-10 of the F1 hybrids between S. officinarum and S. spontaneum. Then, S. spontaneum-specific repetitive DNA probes were used to distinguish S. spontaneum in these hybrids. This oligo-fluorescence in situ hybridization (FISH) system proved to be an efficient tool for revealing individual chromosomal inheritance during nobilization. We discovered the complete doubling of S. officinarum-derived chromosomes in most F1 hybrids. Notably, we also found defective S. officinarum-derived chromosome doubling in the F1 hybrid Yacheng75-4191, which exhibited 1.5n transmission for all nonhomologous chromosomes. Altogether, these results highlight the presence of variable chromosome transmission in nobilization between S. officinarum and S. spontaneum, including 1.5n + n and 2n + n. These findings provide robust chromosome markers for in-depth studies into the molecular mechanism underlying chromosome doubling during the nobilization, as well as tracing chromosomal inheritance for sugarcane breeding. [ABSTRACT FROM AUTHOR]

The yellow vein mosaic disease (YVMD) transmitted through whitefly is a serious threat to okra fruit and quality production. Although, the disease can be managed up to some extent through chemicals by controlling the vector population. But, the use of resistant varieties is most economic and environment friendly. Therefore, the present study was planned to find out the new resistant source through artificial screening against YVMD. For this, the sixteen accessions belonging to seven different okra wild species along with two cultivars of cultivated okra (Pusa Sawani and Punjab Padmini) were evaluated against YVMD during rainy season of 2016 and 2017. Screening against YVMD was conducted using viruliferous whitefly (Bemisa tabaci Genn.) mediated mass inoculation and observations were recorded from symptomless to very severe infection. Observations were recorded consecutively for two years that confirmed no symptoms of YVMD up to 90 Days After Inoculation (DAI) for accession SBT-12597 and SBT-12592 of Hibiscus acutetus resulting zero coefficient of infection (CI) termed as highly resistant. Accessions IC-203833 and IC-470751 of A. angulosus depicting lowest CI value of 2.3 and 3.7 respectively were also assigned as highly resistant to YVMD at 90 DAI. The accession EC-329394 of A. manihot recorded CI value of 67.9 followed by accession IC-141056 (CI 61.6) of A. moschatus and A. tuberculatus (CI = 44.4) were found susceptible to YVMD up to 90 DAI. The cultivated okra A. esculentus variety Pusa Sawani with CI value of 75.0 and Punjab Padmani (CI 60.0) showed highly susceptible and susceptible reaction to YVMD, respectively. Deng universal primer confirmed the resistance in all the selected plants by showing absence of viral DNA. [ABSTRACT FROM AUTHOR]