Fertile Hybrids Research Articles

Changes of hemoglobin resulted by distant hybridization contributed to increased hypoxia tolerance of allotriploid crucian carp

Hypoxia tolerance, one of the important goals of fish breeding, is beneficial for aquaculture and the transporting of fish. Based on the bisexual fertile allotetraploid crucian carp hybrid lineage obtained by distant hybridization, we successfully developed the allotriploid crucian carp via interploidy crossing with diploid red crucian carp. This allotriploid fish exhibited obvious hypoxia tolerance with lower values of dissolved oxygen at Pcrit (critical oxygen press) and 50 % ASR (aquatic surface respiration) than its original parents. Hemoglobin is the main vector of oxygen in vivo. This study examined the changes in amino acid sequences and natural protein conformation of hemoglobin, and found that the Hbα sequences of allotriploid crucian carp displayed obvious hybridity, including recombination and variation, when compared with its original parents. Moreover, the expression of hbα gene is higher in the liver and blood of allotriploid crucian carp than in its original parents. After acute anoxia treatment, the expression of gata1 gene, which served as the general switch for erythroid development, decreased in the blood of all the three kinds of fish. However, only in the allotriploid crucian carp hbα gene expression significantly decreased, coupled with the erythrocytes displaying an increased ratio of nuclei/cytoplasm. Taken together, this study suggested that changes in hemoglobin coupled with the erythrocyte morphological alterations under hypoxia increased the hypoxia tolerance ability of allotriploid crucian carp, which would provide a new perspective to guide fish breeding to obtain high-quality fish species with increased hypoxia tolerance.

Studies on Distant Hybridization Compatibility between the Azalea (Rhododendron × hybridum hort.) and the Rhododendron decorum Franch. Native to China

Rhododendron resources are abundant in China, and hybridization breeding technology is the primary method for cultivating Rhododendron varieties. In order to optimize the utilization of wild Rhododendron resources for distant hybridization, this study took three horticultural varieties of Rhododendron subgenus Tsutsusi and the Rhododendron decorum Franch. of the subgenus Hymenanthes as the research objects, and the cross-compatibility between subgenera was analyzed from the aspects of pollen tube growth and ovary and seed development. At the same time, the statistics of ovary swelling rate and fruit bearing rate, numbers of capsule seeds, 1000 seed weight of hybrid seeds, germination rate, green seedling rate, and other indexes were analyzed to comprehensively evaluate hybrid fertility. The results showed that there was obvious pre-fertilization and post-fertilization barriers existing in the hybridization of Rhododendron × hybridum hort. and R. decorum. During the growth of pollen tubes, a large amount of callus appeared, which led to the entanglement, distortion, and abnormal development of the pollen tubes; only part of the pollen tubes entered into the ovary or ovule. The pre-fertilization barriers can be overcome by early pollination and delayed pollination. According to the observation of the ovary paraffin section, 45 d after pollination, the seed was shriveled and developed abnormally. The comprehensive evaluation of hybrid fertility showed that ‘Sima’ × R. decorum was fertile, ‘Yin Taohong’ × R. decorum was a weakly fertile, and ‘Little Taohong’ × R. decorum was sterile. This study provided a scientific basis for intergeneric hybridization breeding between the subgenus Tsutsusi and the subgenus Hymenanthes.

Sharp decline in male fertility in F2 hybrids of the female-heterogametic silk moth Bombyx.

Sexual selection drives rapid evolution of morphological, physiological, and behavioral traits, especially in males, and it may also drive the rapid evolution of hybrid male sterility. Indeed, the faster male theory of speciation was once viewed as a major cause of Haldane's rule in male-heterogametic XY taxa, but is increasingly being replaced by the genetic conflict hypothesis partly because it cannot explain the faster evolution of hybrid female sterility in female-heterogametic ZW taxa. The theory nonetheless predicts that there should be more genes for hybrid male sterility than for hybrid female sterility even in such taxa, but this remains untested. Thus, finding evidence for the faster male theory of reproductive isolation beyond the F1 generation in ZW systems still represents a challenge to studying the impact of sexual selection. In this study, we examined F2 hybrids between the domesticated silkworm Bombyx mori and the wild silk moth Bombyx mandarina, which have ZW sex determination. We found that although only females showed reduced fertility in the F1 generation, the F2 hybrid males had a significant reduction in fertility compared with the parental and F1 males. Importantly, 27% of the F2 males and 15% of the F2 females were completely sterile, suggesting the presence of recessive incompatibilities causing male sterility in female-heterogametic taxa.

Effects of plant hormones and genotypes on anther culture response of safflower (Carthamus tinctorius L.)

Safflower (Carthamus tinctorius L.) belongs to the composite family with various uses, from the stem to the seeds. In vitro flowering and embryo rescue are common techniques for improving safflower hybrid fertility by overcoming breeding constraints. However, anther culture expedites the production of superior types. The aim of the study was to evaluate the effect of plant hormones and genotypes on anther culture responses and to develop a suitable protocol. In both safflower genotypes, callus, shoot, and root formation were investigated using ten factorial hormone treatments. To promote callus induction and shoot regeneration, variable doses of Thidiazuron (TDZ) at 1 mg/l indole butyric acid (IBA) and variable benzylaminopurine (BAP) at 0.5 mg/l NAA were added to the MS medium, respectively. ½ MS media with varying IBA concentrations at a fixed 0.5 mg/l NAA were used for root regeneration. Results showed that Turkan was superior in all aspects of callus induction, and the highest degree of callus formation (46 %) was observed at 0.5 mg/l TDZ. On the other hand, the local safflower performed better for shoot and root regeneration. The highest shoot regeneration capacity (20 %) was shown at 2.0 mg/l BAP, while shoot rooting was reached (21 %) at 1.0 mg/l IBA of culture media. Despite successful in vitro regeneration, acclimated plantlets did not survive in the glasshouse. In this study, we demonstrated the higher capability for both shoot and root regeneration of calli produced from local safflower anthers. More effort is required to improve shoot and root development for both genotypes.

Tetraploid interspecific hybrids between Asian and African rice species restore fertility depending on killer-protector loci for hybrid sterility.

Interspecific F1 hybrids between Asian (Oryza sativa) and African rice (Oryza glaberrima) exhibit severe sterility caused by the accumulation of hybrid sterility genes/loci at 15 or more loci. The mechanisms underlying the hybrid sterility genes are largely unknown; however, a few genes associated with the killer-protector system, which is the system most frequently associated with hybrid sterility genes, have been identified. We previously produced fertile plants as tetraploids derived from diploid interspecific F1 hybrids through anther culture; therefore, it was suggested that hybrid sterility could be overcome following tetraploidization. We investigated whether tetraploid interspecific plants produced by crossing are fertile and tested the involvement of hybrid sterility genes in the process. Fertile tetraploid interspecific F1 hybrid plants were obtained by crossing 2 tetraploids of O. sativa and O. glaberrima. To elucidate the relationships between pollen fertility and the hybrid sterility loci in the tetraploid F1 microspores, we performed genetic analyses of the tetraploid F2 hybrids and diploid plants obtained from the microspores of tetraploid interspecific hybrids by anther culture. The result suggested that the tetraploid interspecific hybrids overcame pollen and seed infertility based on the proportion of loci with the killer-protector system present in the tetraploids. The heterozygous hybrid sterility loci with the killer-protector system in the tetraploid segregate the homozygous killed allele (16.7-21.4%), with more than three-quarters of the gametes surviving. We theoretically and experimentally demonstrated that fertile rice progenies can be grown from tetraploid interspecific hybrids.

Detection of reproductive interference between closely related Salvia species with small-scale separated distributions by multifaceted pollination and molecular analyses

Reproductive interference, an interspecific interaction in reproductive process that exerts an adverse effect, has gained attention as a contributing factor in promoting exclusive distributions between closely related species. However, detailed studies on the possibility of reproductive interference between native plants are still lacking, presumably because strong reproductive interference can rapidly realize exclusive distributions, leaving the two species apparently independent. Salvia japonica and S. lutescens are found in separate localities at a small scale, although their distributions overlap at a large scale. We investigated the possibility of reproductive interference between them through field surveys, hand-pollination experiments, evaluation of hybrid fertility, cpDNA and nrDNA genotyping, and genome-wide DNA analysis. The field survey results did not reveal apparent negative interaction in competition for pollinator services. Mixed pollination with conspecific pollen and counterpart pollen reduced seed set in S. japonica, and hybrid progeny produced by mixed pollination were less than 20% as fertile compared to the pure species. The DNA genotyping results suggested the possibility of hybridization where their distributions overlap, and the genome-wide DNA analysis results showed clear genetic differentiation between the two species as well as the existence of hybrids. These results suggest that bi-directional reproductive interference between S. japonica and S. lutescens may have led to their present separated distributions at a small scale.

Assessment of biological and productive parameters of female hybrids between the Volga sterlet (Acipenser ruthenus) and Amur kaluga (Huso dauricus). Part 2. Hybrids between kaluga females and sterlet males. Comparative characteristics of reciprocal hybrids

The second part (for the first part of the article, see No. 6, 2024) of the work describes the production indicators of fertile female hybrids between Kaluga females and sterlet males, and also provides a comparison with the previously described indicators of hybrids between sterlet females and Kaluga males. The study of the production parameters of reciprocal hybrids in the process of long-term formation of breeding stocks is of both scientific and practical interest. The purpose of the work is to evaluate the fish-breeding, biological and production characteristics of female hybrids, and the effectiveness of their use as promising objects for obtaining food caviar. The maturation periods, inter-spawning intervals, the weight of the eggs obtained and the weight of the eggs, fertility and oosomatic indices of 67 females of 5 age-related generations of reciprocal hybrids from the warmwater farm of the Primorsky Territory weighing from 17 to 35 kg, participating in 87 spawning campaigns from the age of 9 to 14–18 years, were studied. One of the generations is represented by real albinos of lemon-yellow color with red eyes and light caviar. The conducted studies have shown that the number of fertile females in different generations varies from 10 to 30%, but from 25% of females it was not possible to obtain eggs even once. Females begin to mature at the age of 9–12 years, the duration of maturation of all females of the same generation reaches 8–10 years. Most females skip 2–4 years between spawning or mature once. Caviar resorption in winter occurs in 17–42% of female hybrids. On average, during one spawning, females produce from 1,5 to 1,7 kg of eggs with an average weight of 23 mg of eggs. The production performance of hybrids is significantly lower compared to hybrids between beluga and sterlet (besters). Due to their biological characteristics and low productivity, reciprocal hybrids between Kaluga and sterlet are of greater scientific interest for genetic research than as producers of caviar. The cultivation of hybrids in order to obtain industrial volumes of caviar for food purposes is recognized as costly, ineffective and inexpedient.

Usefulness of alien sterilizing cytoplasms for the hybrid breeding of triticale (xTriticosecale Wittmack): preliminary results.

To be useful for cereal breeding, cytoplasmic male sterility (CMS) should express the complete sterility of maternal lines and the full restoration of the male fertility of F1 hybrids. The most reliable source of sterilizing cytoplasm for triticale is Triticum timopheevi; however, due to the low frequency of efficient non-restorer genotypes for this cytoplasm, new sources of CMS are needed. In this study, aside from T. timopheevi (T) cytoplasm, three alternative CMS sources were tested: Pampa (P) from Secale cereale L., Aegilops sharonensis (A), and Ae. ventricosa (V). The suitability of these cytoplasms for breeding was assessed based on the male fertility/sterility of F1 hybrids obtained through the manual pollination of CMS maternal lines with 36 triticale cultivars and breeding strains. About half of the hybrids with each type of cytoplasm were fully fertile and produced more than 30 grains per bagged spike. The highest percentage was found in hybrids with P cytoplasm (58.33%) and the lowest in hybrids with A cytoplasm (44.44%). Male sterility was observed in hybrids with P cytoplasm (16.67%) and A cytoplasm (16.67%) but not in hybrids with T or V cytoplasm. In terms of practical aspects, male sterility systems with P or A cytoplasm exhibit similarity in their ability to restore male fertility that differ from the T and V cytoplasms. Although all studied cytoplasms exhibited some disadvantages for breeding purposes, none should be definitively classified as unacceptable for future breeding programs regarding the development of triticale hybrid cultivars.

Asymmetric Introgression and Cryptic Natural Hybridization between Two Species of Teucrium Section Polium (Lamiaceae) on the Balkan Peninsula.

In this work, we analyzed the morphology and genetic structure of Teucrium montanum, T. capitatum and their hybrid T. × rohlenae from three syntopic populations. A morphometric study showed that the parents and their hybrids exhibited continuous morphological variation, with the hybrid positioned exactly between the parents. Genetic analysis revealed that plants morphologically identified as T. × rohlenae are fertile hybrids that produce hybrid swarms dominated by later-generation hybrids. This suggests that introgression, rather than speciation, is the more likely outcome of hybridization between these plant species. The extent and direction of gene flow between the two species differed markedly between the three syntopic localities. At the Trilj locality, it was clearly unidirectional, with T. capitatum playing the dominant role. At the Sićevo locality, gene flow was slightly asymmetric, favoring the genetic background of T. capitatum, while at the Sliven site, it was completely asymmetric in the opposite direction. The extreme case of unidirectional gene flow was observed at the Trilj locality where plants morphologically identified as T. montanum could not be genetically distinguished from T. capitatum. This suggests that interspecific hybridization occurred long ago, leading to introgression and cryptic hybrids, blurring of species boundaries and generating evolutionary noise.

North American pitseed goosefoot (Chenopodium berlandieri) is a genetic resource to improve Andean quinoa (C. quinoa)

Pitseed goosefoot (Chenopodium berlandieri) is a free-living North American member of an allotetraploid complex that includes the Andean pseudocereal quinoa (C. quinoa). Like quinoa, pitseed goosefoot was domesticated, possibly independently, in eastern North America (subsp. jonesianum) and Mesoamerica (subsp. nuttaliae). To test the utility of C. berlandieri as a resource for quinoa breeding, we produced the whole-genome DNA sequence of PI 433,231, a huauzontle from Puebla, México. The 1.295 Gb genome was assembled into 18 pseudomolecules and annotated using RNAseq data from multiple tissues. Alignment with the v.2.0 genome of Chilean-origin C. quinoa cv. ‘QQ74’ revealed several inversions and a 4A-6B reciprocal translocation. Despite these rearrangements, some quinoa x pitseed goosefoot crosses produce highly fertile hybrids with faithful recombination, as evidenced by a high-density SNP linkage map constructed from a Bolivian quinoa ‘Real-1’ × BYU 937 (Texas coastal pitseed goosefoot) F2 population. Recombination in that cross was comparable to a ‘Real-1’ × BYU 1101 (Argentine C. hircinum) F2 population. Furthermore, SNP-based phylogenetic and population structure analyses of 90 accessions supported the hypothesis of multiple independent domestications and descent from a common 4 × ancestor, with a likely North American Center of Origin.

Contribution to the Embryology, Sporoderm and Spermoderm Pattern of <I>Bothriochloa pertusa</I> (L.) A. Camus, <I>Capillipedium filiculme</I> (Hook.F.) Stapf and <I>Dichanthium annulatum</I> (Forssk.) Stapf with a Brief discussion on its Taxonomic Status

Embryological features in Bothriochloa pertusa, Capillipedium filiculme and Dichanthium annulatum have been studied. The male reproductive characters are constant as evidenced by anther wall development and it corresponds to Monocot type. At maturity the pollen grains are 3-celled. Embryo sac development and embryogeny conforms to Polygonum type and Asterad type respectively. Endosperm, seed coat and pericarp have been studied and data on SEM investigation on pollen wall and seed coat of these taxa are added. The three taxa viz., Bothriochloa, Capillipedium and Dichanthium were considered under Andropogoneae of Panicoideae as separate genera by Bor (1960). De Wet and Harlan (1970) merged them in Dichanthium . The evidences given by them are intergeneric hybridization and production of fertile hybrids between these three genera. Clayton (1977) also followed the concept of merger taking priority. The authors assessed the possibility of merger into Dichanthium on the basis of embryology, sporoderm and spermoderm pattern

Genome Variability in Artificial Allopolyploid Hybrids of Avena sativa L. and Avena macrostachya Balansa ex Coss. et Durieu Based on Marker Sequences of Satellite DNA and the ITS1-5.8S rDNA Region.

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.

Assessment of Transgressive Segregants for Yield and its Component Traits in French Bean (Phaseolus vulgaris L.)

Background: Extreme phenotypes carried on by transgressive segregation are, moreover, heritably stable in contrast to heterosis. It has been hypothesized that the development of extreme or “transgressive” traits in segregating hybrid populations assists in the adaptive divergence of hybrid taxa. The investigation on French bean was experimented to evaluate transgressive segregants generated in the F2 generation for yield and related traits. Methods: The present study on French bean was conducted during rabi 2019 to 2022 to elicit data on transgressive segregants produced for yield and related traits in F2 generation. Fertile F1 hybrids were developed from three crosses: IC 632961 × Arka Sukomal, IC 632961 × Arka Arjun and Arka Arjun × IC 63296. Heterosis was estimated for yield and related traits in these hybrids. The F2 progeny were evaluated in open field condition. Mean, standard deviation and percentage of transgressive segregants were estimated for quantitative traits under investigation. Result: Transgressive segregants were observed in all three crosses, with the highest number of transgressive segregants for yield traits found in the progeny of the cross IC 632961 × Arka Sukomal. The identification and isolation of such transgressive segregants indicate that the parents possess favourable alleles and genes governing the traits of interest, which can be consolidated into a single or a few genotypes through stringent selection. Superior and desirable transgressive segregants were observed for all sixteen yield and its component traits, except for leaf length, in the two cross combinations Arka Arjun × IC 632961 and IC 632961 × Arka Arjun. The highest percentage of transgressive segregation was observed for the number of inflorescences per plant (79.49%), followed by the number of pods per plant (78.46%) and pod yield per plant (75.61%) in the F2 generation.

Possibilities of using the tetraploid interspecific hybrid <i>Hordeum bulbosum</i> L. × <i>Hordeum vulgare</i> L. in obtaining new recombinant barley lines

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.

Elevated ozone disrupts mating boundaries in drosophilid flies

Animals employ different strategies to establish mating boundaries between closely related species, with sex pheromones often playing a crucial role in identifying conspecific mates. Many of these pheromones have carbon-carbon double bonds, making them vulnerable to oxidation by certain atmospheric oxidant pollutants, including ozone. Here, we investigate whether increased ozone compromises species boundaries in drosophilid flies. We show that short-term exposure to increased levels of ozone degrades pheromones of Drosophila melanogaster, D. simulans, D. mauritiana, as well as D. sechellia, and induces hybridization between some of these species. As many of the resulting hybrids are sterile, this could result in local population declines. However, hybridization between D. simulans and D. mauritiana as well as D. simulans and D. sechellia results in fertile hybrids, of which some female hybrids are even more attractive to the males of the parental species. Our experimental findings indicate that ozone pollution could potentially induce breakdown of species boundaries in insects.

The genomic signatures of evolutionary stasis.

Evolutionary stasis characterizes lineages that seldom speciate and show little phenotypic change over long stretches of geological time. Although lineages that appear to exhibit evolutionary stasis are often called living fossils, no single mechanism is thought to be responsible for their slow rates of morphological evolution and low species diversity. Some analyses of molecular evolutionary rates in a handful of living fossil lineages have indicated that these clades exhibit slow rates of genomic change. Here, we investigate mechanisms of evolutionary stasis using a dataset of 1,105 exons for 481 vertebrate species. We demonstrate that two ancient clades of ray-finned fishes classically called living fossils, gars and sturgeons, exhibit the lowest rates of molecular substitution in protein-coding genes among all jawed vertebrates. Comparably low rates of evolution are observed at fourfold degenerate sites in gars and sturgeons, implying a mechanism of stasis decoupled from selection that we speculate is linked to a highly effective DNA repair apparatus. We show that two gar species last sharing common ancestry over 100 million years ago produce morphologically intermediate and fertile hybrids in the wild. This makes gars the oldest naturally hybridizing divergence among eukaryotes and supports a theoretical prediction that slow rates of nucleotide substitution across the genome slow the accumulation of genetic incompatibilities, enabling hybridization across deeply divergent lineages and slowing the rate of speciation over geological timescales. Our results help establish molecular stasis as a barrier to speciation and phenotypic innovation and provide a mechanism to explain the low species diversity in living fossil lineages.

Hybridization and gene expression: Beyond differentially expressed genes.

Gene expression has a key role in reproductive isolation, and studies of hybrid gene expression have identified mechanisms causing hybrid sterility. Here, we review the evidence for altered gene expression following hybridization and outline the mechanisms shown to contribute to altered gene expression in hybrids. Transgressive gene expression, transcending that of both parental species, is pervasive in early generation sterile hybrids, but also frequently observed in viable, fertile hybrids. We highlight studies showing that hybridization can result in transgressive gene expression, also in established hybrid lineages or species. Such extreme patterns of gene expression in stabilized hybrid taxa suggest that altered hybrid gene expression may result in hybridization-derived evolutionary novelty. We also conclude that while patterns of misexpression in hybrids are well documented, the understanding of the mechanisms causing misexpression is lagging. We argue that jointly assessing differences in cell composition and cell-specific changes in gene expression in hybrids, in addition to assessing changes in chromatin and methylation, will significantly advance our understanding of the basis of altered gene expression. Moreover, uncovering to what extent evolution of gene expression results in altered expression for individual genes, or entire networks of genes, will advance our understanding of how selection moulds gene expression. Finally, we argue that jointly studying the dual roles of altered hybrid gene expression, serving both as a mechanism for reproductive isolation and as a substrate for hybrid ecological adaptation, will lead to significant advances in our understanding of the evolution of gene expression.

Reproductive relationships between taxa morphologically close to Elymus caninus (Poaceae: Triticeae).

A hybridological study of biotypes of species close to Elymus caninus: E. prokudinii, E. viridiglumis, E. goloskokovii, as well as a number of morphologically deviant biotypes in Russia and Kazakhstan, was carried out. The objectives were to study the levels of reproductive relationships and the degree of integration of the species E. goloskokovii, E. prokudinii, and E. viridiglumis into the E. caninus complex. Our estimates of the seed fertility of natural parental biotypes were within 60-90 %. Among the combinations of crossing in F1, the highest seed setting was found in the hybrids formed by parental pairs from close habitats, regardless of the taxonomic rank of biotypes. The highest fertility values (55.6 and 46.1 %) were found in combinations involving E. caninus, E. viridiglumis and E. goloskokovii. It has been concluded that the biotypes of these species included in sexual hybridization form a single recombination gene pool, within which slight differences in reproductive compatibility are observed. The nature of the inheritance of the diagnostic features of lemmas "presence of trichomes" and "length of awns", according to the digenic and monogenic type, respectively, is shown. The high seed fertility of the created hybrids and the presence of intermediate forms in the F2 generation according to distinctive features indicate the possibility of interspecific introgression when species grow together in natural populations. Thus, the assessment of the inheritance of diagnostic characters makes it possible to classify E. goloskokovii, E. prokudinii, and E. viridiglumis as intraspecific taxa of E. caninus s. l. Data were obtained on the morphological and reproductive properties of interspecific hybrids with the participation of the species E. mutabilis as a possible donor in the speciation of taxa close to E. caninus. In cross combinations of E. caninus × E. mutabilis and E. mutabilis × E. caninus, lower values of seed fertility of hybrids in the F1 and F2 generations were noted compared to hybrids between the species E. caninus, E. goloskokovii, E. prokudinii and E. viridiglumis. Nevertheless, on the basis of chorological and morphological criteria, we concluded that E. caninus and E. mutabilis are independent species.

Slow-release hydroxyapatite fertilizer from crab shells waste for sustainable crop production

Slow-release fertilizer (SRF) is an alternative to more efficient fertilization. SRF can gradually release nutrients in fertilizer, hence the nutrients needed by for the plant growth will be available for a more extended period. In this study, SRF was made by mixing single fertilizer (urea and ammonium sulfate) with hydroxyapatite particles from crab shell biowaste. Slow-release hydroxyapatite fertilizer synthesized with a ratio between single fertilizer and hydroxyapatite powder of 6:1, then applied to grow Capsicum annuum pepper plants. The result showed that applying hybrid fertilizer affected the growth and yield of Capsicum annuum pepper plant, by enhancement of plant height up to 150 %, stem diameter up to 168 %, number of leaves up to 167 %, number of fruits up to 449 %, fruit diameter up to 200 % and fruit weight up to 234 %, compared with unfertilized plant. The SRF fertilization slightly affect the soil acidity, however the pH changed showed no effect to the plant growth. The use of HA as fertilizer promotes more sustainable crop production, by efficient nutrient uptake and utilization calcium-rich biowaste.

Editing of ORF138 restores fertility of Ogura cytoplasmic male sterile broccoli via mitoTALENs.

Cytoplasmic male sterility (CMS), encoded by the mitochondrial open reading frames (ORFs), has long been used to economically produce crop hybrids. However, the utilization of CMS also hinders the exploitation of sterility and fertility variation in the absence of a restorer line, which in turn narrows the genetic background and reduces biodiversity. Here, we used a mitochondrial targeted transcription activator-like effector nuclease (mitoTALENs) to knock out ORF138 from the Ogura CMS broccoli hybrid. The knockout was confirmed by the amplification and re-sequencing read mapping to the mitochondrial genome. As a result, knockout of ORF138 restored the fertility of the CMS hybrid, and simultaneously manifested a cold-sensitive male sterility. ORF138 depletion is stably inherited to the next generation, allowing for direct use in the breeding process. In addition, we proposed a highly reliable and cost-effective toolkit to accelerate the life cycle of fertile lines from CMS-derived broccoli hybrids. By applying the k-mean clustering and interaction network analysis, we identified the central gene networks involved in the fertility restoration and cold-sensitive male sterility. Our study enables mitochondrial genome editing via mitoTALENs in Brassicaceae vegetable crops and provides evidence that the sex production machinery and its temperature-responsive ability are regulated by the mitochondria.