Chromosome Behavior Research Articles

Taming wild genomes: perspectives on mechanisms governing differential introgression in exotic rice germplasm and their applications in breeding

Wide hybridization is an important plant breeding strategy that can be used to expand the available genetic variation in present-day crops towards breeding for enhanced agronomic performance. The primary challenge in wide hybridization is the presence of reproductive barriers and genetic incompatibilities that limit the transfer of desirable wild or distant alleles in the genetic background of cultivated plant species. Here we provide perspectives on the possible role of hybrid sterility and gametocidal genes on the observed preferential introgression in exotic germplasm of rice. We argue that while these aberrant introgression and segregation behavior of wild or distant chromosomes presents significant barriers in exploiting ancestral germplasm in breeding, the same mechanisms can also be exploited to enhance the transfer of wild alleles in a cultivated genetic background. Understanding the genetic basis of preferential introgression and segregation in wide hybrids will have serious implications in our ability to capture ancestral genetic variation that can add significant agronomic value to staple crops like rice.

Development and characterization of a complete set of monosomic alien addition lines from Raphanus sativus in Brassica oleracea.

A complete set of monosomic alien addition lines of Radish-Brassica oleracea exhibiting extensive variations was generated and well characterized for their chromosome behaviors and phenotypic characteristics. Monosomic alien addition lines (MAALs) are developed through interspecific hybridization, where an alien chromosome from a relative species is introduced into the genome of the recipient plant, serving as valuable genetic resources. In this study, an allotetraploid Raphanobrassica (RRCC, 2n = 36) was created from the interspecific hybridization between radish (Raphanus sativus, RR, 2n = 18) and Brassica oleracea (CC, 2n = 18). Subsequently, this Raphanobrassica was repeatedly backcrossed with radish to generate an aneuploid population. The identification of a complete set of MAALs (RR + 1C1-9, 2n = 19) was achieved using PCR with C chromosome-specific markers and fluorescence in situ hybridization, revealing extensive morphological variations, particularly in the shape and size of the fleshy root. A complete set of MAALs was achieved with only one chromosome from 1 to 9 linkage groups of the C genome. Compared with parental radish, most of the MAALs showed a noticeable delay in root swelling, particularly the RR-C6 that did not exhibit obvious root swelling throughout its entire growth stage. Cytological analysis indicated that the MAAL lines containing chromosome C8 exhibited the highest frequency of intergenomic chromosome pairings. Additionally, some introgressive radish lines derived from MAALs displayed a preference toward the donor B. oleracea or over-parent heterosis for some certain nutritional components. Overall, these MAALs serve as valuable germplasm for the genetic enhancement of radish and provide insights into the interactions between the R genome and C chromosomes.

Development, characteristics, and gene expression profiles of a chromosomal deletion line of Brassica juncea (AABB, 2n=36) with apetalous feature derived from interspecific hybridization

Interspecific hybridization has been a crucial technique in improving genetic diversity and developing novel traits in cultivated plants. The apetalous Brassica juncea (AABB, 2n=36) represents a valuable germplasm resource with highly desirable characteristics for breeding purposes. Nevertheless, apetalous germplasm resources in B. juncea are currently scarce. In this study, we successfully established a stable apetalous B. juncea (ABJ) line by crossing B. juncea “YLC” with B. carinata “Bc216” (BBCC, 2n=34). Cytological analysis indicates that ABJ exhibited normal chromosome behavior, with 18 bivalents in diakinesis and an equal number of chromosome segregations at anaphase I in the pollen mother cells during meiosis. Fluorescence in situ hybridization (FISH) confirmed that the chromosome configuration of ABJ is similar to that of its parental line B. juncea, containing 20 chromosomes from A-subgenome and 16 chromosomes from B-subgenome constituting the chromosome number of 36. However, a 15.8 Mb deletion in chromosome A03 of ABJ was uncovered by RNA-seq and molecular markers. Twenty-four differentially expressed genes which may be associated with the regulation of ABJ's apetalous traits were identified by gene expression profiling. To our knowledge, there have been no other documented instances of producing apetalous mustard with missing chromosome segments through interspecific hybridization. Consequently, this study not only verifies the feasibility of generating apetalous mustard with missing chromosome segments through interspecific hybridization but also provides a new insight into the mechanisms underlying the formation of apetalous trait.

Varied chromosome distribution behaviours during meiosis in triploid Chinese chives contribute to the formation of viable pollen.

Triploids play an important role in the polyploidization process and are considered a bridge between diploids and polyploids. To inform plant polyploidization research and polyploid breeding, it is important to explore chromosome behaviour during triploid pollen development, pollen fertility problems in triploids and the potential value of utilizing triploids. In this study, acetocarmine, carbol fuchsin and fluorescence staining methods were used to observe microsporogenesis and microspore development in fertile triploid Chinese chives. The results revealed that some of the pollen mother cells were able to undergo equal chromosome distributions (approximately 36%), whereas other pollen mother cells formed lagging chromosomes, chromosome bridges, micronuclei and early cytoplasmic divisions during microsporogenesis, resulting in microspores of different sizes. Regardless of whether an equal tetrad or an abnormal polyad was formed, microspores were released from callose in a normal manner and contained nuclei. During the process of microspore development, most of the microspore nuclei disappeared gradually and ultimately formed empty pollen cells that lacked nuclei. During the meiosis of pollen mother cells in triploid Chinese chives, a variety of chromosome distribution behaviours contribute to the formation of some viable pollen.

RMI1 is essential for maintaining rice genome stability at high temperature.

Heat is a critical environmental stress for plant survival.One of its harmful effects on the cells is the disruption of genome integrity. However, the mechanisms by which plants cope with heat-induced DNA damage remain largely unknown. RMI1, a component of the RTR (RECQ4-TOP3α-RMI1) complex, plays a pivotal role in maintaining genome stability. In this study, we identified the target gene RMI1by characterizing a high-temperature-sensitive mutant. The growth and development of rmi1-1 seedlings carrying a non-frameshift mutation in RMI1 were hindered at 38°C. Abnormal mitotic chromosome behaviours ultimately led to the cell death of root tips. Additionally, the presence of chromosome fragments during anaphase I caused pollen abortion and sterility in rmi1-1 plants. Yeast two-hybrid assays revealed that the interactions between RMI1-1 and RECQ4 or TOP3α were weakened with increasing temperature and entirely ceased at 36°C. In contrast, the functionalRMI1 maintained its interactions with RECQ4 or TOP3α under the same conditions. These results indicate that the non-frameshift mutation in RMI1 disrupts the formation of the RTR complex at high temperatures, leading to defects in DNA repair and increased sensitivity of rmi1-1 under heat stress. However, embryos of the rmi1-cr2 mutant with a frameshift mutation in RMI1 exhibited complete lethality. In addition, the overexpression of RMI1 enhanced the heat tolerance in rice. These findings provide insights into the molecular mechanisms that RMI1 responds to high temperatures by maintaining genome stability in rice.

The Cytological Mechanism of the Peach Haploid Producing Triploid Offspring

Abstract Peach is one of the most economically valuable fruit trees. Haploid peach trees occur spontaneously at very low frequencies and they are usually highly sterile. Therefore, the haploid with partial fertility is extremely rare germplasm, which is highly valuable to genetic research and breeding programmes. In this study, we investigated the cytological mechanism underlying the fertility of a peach haploid mutant ‘9-D’ derived from a spontaneous mutation. Cytologic evaluation and flow cytometry analysis demonstrated that ‘9-D’ is a pure haploid. Scanning electron microscope analysis revealed a considerable proportion of abnormal pollen grains in ‘9-D’. Pollen viability assay by Alexander staining showed that 50.4% of pollen grains from ‘9-D’ were viable. However, the pollen germination assay showed that only 7.6% of the pollen grains could germinate normally. Investigation of the chromosomal behavior of pollen mother cells at different stages of meiosis showed that pollen mother cells of ‘9-D’ lacked the process between anaphase I and prophase II of meiosis. Various types of sporophyte morphology were observed in haploid pollen mother cells at the tetrad stage. Measurement of the diameter of pollen grains indicated the presence of pollen with 2x ploidy in ‘9-D’. The offspring of ‘9-D’ were predominantly triploid or triploid aneuploid. The triploid offspring were more likely derived from the 2x male gametophyte combined with the haploid female gametophyte, which may explain the reason why ‘9-D’ has fertility. This study not only expands our understanding of haploid fertility mechanisms, but is also useful for ploid breeding programs in peach.

Unambiguous chromosome identification reveals the factors impacting irregular chromosome behaviors in allotriploid AAC Brassica.

The major irregular chromosome pairing and mis-segregation were detected during meiosis through unambiguous chromosome identification and found that allotriploid Brassica can undergo meiosis successfully and produce mostly viable aneuploid gametes. Triploids have played a crucial role in the evolution of species by forming polyploids and facilitating interploidy gene transfer. It is widely accepted that triploids cannot undergo meiosis normally and predominantly produce nonfunctional aneuploid gametes, which restricts their role in species evolution. In this study, we demonstrated that natural and synthetic allotriploid Brassica (AAC), produced by crossing natural and synthetic Brassica napus (AACC) with Brassica rapa (AA), exhibits basically normal chromosome pairing and segregation during meiosis. Homologous A chromosomes paired faithfully and generally segregated equally. Monosomic C chromosomes were largely retained as univalents and randomly entered daughter cells. The primary irregular meiotic behaviors included associations of homoeologs and 45S rDNA loci at diakinesis, as well as homoeologous chromosome replacement and premature sister chromatid separation at anaphase I. Preexisting homoeologous arrangements altered meiotic behaviors in both chromosome irregular pairing and mis-segregation by increasing the formation of A-genomic univalents and A-C bivalents, as well as premature sister chromatid separation and homologous chromosome nondisjunction. Meiotic behaviors depended significantly on the genetic background and heterozygous homoeologous rearrangement. AAC triploids mainly generated aneuploid gametes, most of which were viable. These results demonstrate that allotriploid Brassica containing an intact karyotype can proceed through meiosis successfully, broadening our current understanding of the inheritance and role in species evolution of allotriploid.

Abnormal expression of LiASY1 and LiDMC1 is the main cause of sterility in F1 hybrids developed from Lagerstroemia speciosa and Lagerstroemia indica ‘Ebony Embers’

Lagerstroemia × ‘Xiaoxiang Huaxiu’ (an F1 hybrid of Lagerstroemia speciosa and Lagerstroemia indica ‘Ebony Embers’) exhibits significant sterility. Cytological studies have revealed numerous abnormal chromosomal behaviors during the meiotic division of pollen mother cells in L. × ‘Xiaoxiang Huaxiu’. Transcriptome sequencing identified that LiDMC1 and LiASY1 were expressed at very low levels in ‘Xiaoxiang Huaxiu’. To further verify whether the abnormal expression of LiDMC1 and LiASY1 was the fundamental cause of sterility in ‘Xiaoxiang Huaxiu’, we cloned the full-length gene sequences of LiDMC1 and LiASY1 from ‘Ebony Embers’. Using VIGS (virus-induced gene silencing) technology, pTRV1 and pTRV2 gene silencing vectors were employed to silence LiDMC1 and LiASY1 in the paternal parent ‘Ebony Embers’. The results indicated that in the double-silenced lines of ‘Ebony Embers’, the cytoplasm sizes of different tetrads during the tetrad stage varied significantly. There was a notable proportion of polyads, binucleate cells with micronuclei, and pentads. Additionally, pollen diameter varied significantly, closely resembling the sterility observed in ‘Xiaoxiang Huaxiu’. Pollen viability in the double-silenced ‘Ebony Embers’ decreased by 22.8%. This study demonstrates that the low expression of LiDMC1 and LiASY1 leads to abnormal microspore development in ‘Ebony Embers’, and their low expression is the cause of sterility in ‘Xiaoxiang Huaxiu’.

Integrated cytological and transcriptomic analyses provide new insights into restoration of pollen viability in synthetic allotetraploid Brassica carinata.

Upregulation of genes involved in DNA damage repair and sperm cell differentiation leads to restoration of pollen viability in synthetic allotetraploid B. carinata after chromosome doubling. Apart from the well-known contribution of polyploidy to crop improvement, polyploids can also be induced for other purposes, such as to restore the viability of sterile hybrids. The mechanism related to viability transition between the sterile allodiploid and the fertile allotetraploid after chromosome doubling are not well understood. Here, we synthesised allodiploid B. carinata (2n = 2x = 17) and allotetraploid B. carinata (2n = 4x = 34) as models to investigate the cytological and transcriptomic differences during pollen development. The results showed that after chromosome doubling, the recovery of pollen viability in allotetraploid was mainly reflected in the stabilisation of microtubule spindle morphology, normal meiotic chromosome behaviour, and normal microspore development. Interestingly, the deposition and degradation of synthetic anther tapetum were not affected by polyploidy. Transcription analysis showed that the expression of genes related to DNA repair (DMC1, RAD51, RAD17, SPO11-2), cell cycle differentiation (CYCA1;2, CYCA2;3) and ubiquitination proteasome pathway (UBC4, PIRH2, CDC53) were positively up-regulated during pollen development of synthetic allotetraploid B. carinata. In summary, these results provide some refreshing updates about the ploidy-related restoration of pollen viability in newly synthesised allotetraploid B. carinata.

Kinesin-1-like protein PSS1 is essential for full-length homologous pairing and synapsis in rice meiosis.

The pairing and synapsis of homologous chromosomes are crucial for their correct segregation during meiosis. The LINC (Linker of Nucleoskeleton and Cytoskeleton) complex can recruit kinesin protein at the nuclear envelope, affecting telomere bouquet formation and homologous pairing. Kinesin-1-like protein Pollen Semi-Sterility1 (PSS1) plays a pivotal role in male meiotic chromosomal behavior and is essential for fertility in rice. However, its exact role in meiosis, especially as kinesin involved in homologous pairing and synapsis, has not been fully elucidated. Here, we generated three pss1 mutants by genome editing technology to dissect PSS1 biological functions in meiosis. The pss1 mutants exhibit alterations in the radial microtubule organization at pachytene and manifest a deficiency in telomere clustering, which is critical for full-length homologous pairing. We reveal that PSS1 serves as a key mediator between chromosomes and cytoskeleton, thereby regulating microtubule organization and transmitting the force to nuclei to facilitate homologous chromosome pairing and synapsis in meiosis.

Characteristics and Cytological Analysis of Several Novel Allopolyploids and Aneuploids between Brassica oleracea and Raphanus sativus.

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.

Indications of programmed cell death in wheat roots upon exposure to silver nanoparticles

Programmed cell death (PCD) can occur at every developmental stage as a plant’s response to various biotic and abiotic environmental factors. Silver nanoparticles (AgNPs) are widely used in consumer products and possess antimicrobial properties, making them important in assessing nanoparticle effects on plants. In the present study, we examined the impact of AgNPs (0, 0.5, 1, 5, 10, and 20 mg L-1) on wheat root PCD by evaluating parameters such as the mitotic index, chromosomal behaviors, nuclear deformation, cytochrome c release, caspase-1-like activity, and the expression of cysteine protease genes (TaVPE4, TaMCA1, and TaMCA4). Our findings revealed a dose-dependent decrease in the mitotic index ratio and increased chromosomal abnormalities induced by AgNPs. Additionally, we observed various hallmarks of PCD, including chromatin condensation, slight DNA smear, reduction in mitochondrial inner membrane potential, and cytochrome c release to the cytoplasm as well as increased caspase-1-like activity and TaVPE4 gene expression. Notably, the gene expressions of TaMCA1 and TaMCA4 were found to be antagonistically regulated by AgNPs, further indicating the induction of PCD by AgNP treatment. Overall, our study provides evidence of AgNP-induced PCD in wheat roots, elucidating the involvement of cysteine protease genes in this process.

Role of ten different exogenous plant growth promoters in regulating cytotoxic and genotoxic processes in barley exposed to high temperature stress

Agricultural crop affected preliminary and the most prominent by the adverse effects of global climate change have to adapt to various abiotic factors that will occur as a result of climate fluctuations in the near-future and struggle to survive. Among abiotic factors, the one of the greatest impact on plant stress is high temperature. Therefore, the most important step to take action against the global threat is the development of new temperature tolerant varieties. Barley, which is the fourth most important cereal in the world after wheat, maize, and rice are affected by high ambient temperatures. In this work, the effects of their alone and in double, triple combinations of ten various plant growth regulators (PGRs) on mitotic activity and chromosome behaviors in root meristems of barley exposed to high temperature (30°C) were investigated. In the experiments, Hordeum vulgare L. cv. Bülbül 89 variety and GA3, KIN, BA, E, EBR, TRIA and PAs (Spm, Spd, Put, Cad) as growth regulators were used. The results obtained were compared with each other and with those of the seeds germinated at optimum temperature (20°C). Consequently; it has been determined that most of the PGRs studied, especially the GA3 and their combinations with GA3, exhibit a very successful performance on mitotic activity and cytogenetic aberrations in barley seeds germinated under high temperature stress- HTS conditions. The effects of these PGRs (except for EBR) and their combinations on mitotic activity and chromosome behaviors under HTS have been presented in this study for the first time.

Physiological, genetical changes and cdc2 gene expression for osmotic stressed Vicia faba reveal the alleviation effect of gamma radiation and putrescine

Climate change caused increasing in soil salinity worldwide. Therefore, it is critical to enhance the capability of plant to tolerate salinity stress. For this goal, putrescine and irradiation by gamma radiation were used to improve the salt tolerance of Vicia faba (the most important human crop). The results indicated depression in mitotic division and all growth parameters associated with the induction of micronucleus (MN) when salinity increased to 100mM, while there was increase in mitotic aberrations. NaCl decreased total soluble sugar, while total N%, total free amino acid, proline and protein contents showed slight increase with increasing salinity stress. Putrescine and gamma radiation mitigated the effect of salinity on cell division and growth parameters. Salt stress decreased the expression of gene encoding cyclin-dependent kinase 2 (cdc2). Both putrescine and gamma radiation increased cdc2 expression. The genetic diversity has been detected among control and treated V. faba using ISSR and SCoT markers. Ten primers had successfully generated 129 reproducible polymorphic amplicons that were suitable for studying the genetic diversity between studied genotypes. ISSR markers provided more discriminating data and were more informative than SCoT markers. Besides, cluster analysis using UPGMA and PAC successfully explained the genetic diversity within studied genotypes. These findings emphasize the efficiency of putrescine and gamma radiation for alleviating the negative impact of salt stress. Moreover, prove the importance of assessing mitotic activity, chromosomes behavior, physiological parameters, the expression level of cdc2 and molecular diversity in V. faba under stress to improve the salt tolerance of it.

Cytogenetic analysis of meiotic behaviour and stability in a trigeneric hybrid (triticale x trigopiro).

Trigeneric hybrids in Triticeae may help to establish evolutionary relationships among different genomes present in the same cellular genetic background and to transfer different alien characters into cultivated wheat. In the present study, a trigeneric hybrid involving species of Triticum, Secale, and Thinopyrum was synthesized by crossing hexaploid triticale with hexaploid trigopiro. The meiotic behaviour of chromosomes belonging to different genomes was analyzed, using routine and in situ hybridization techniques in F1, F2, and F3 generations of the trigeneric hybrid. The purpose of this study was to determine the chromosome number and genomic constitution and to discuss the mechanisms involved in the stabilization of the artificial tricepiro hybrids. The chromosome number of the trigeneric F1 hybrid was 2n = 42. Between 12 and 16 bivalents were observed in the central zone of the equatorial meiotic plate and between 9 and 18 univalents were found in the periphery of the MI equatorial plate. Seven of these univalents showed hybridization signals with rye DNA. Lagging rye and non-rye chromosomes and separation of sister chromatids were found in anaphase I. Tetrads with a maximum of six micronuclei, with and without hybridization signals of rye DNA, were observed. After three generations, meiotic cells revealed the presence of 42 chromosomes and 21 bivalents in diakinesis cells. The presence of 14 rye (Secale cereale) chromosomes and the complete pairing of chromosomes in F3 hybrids suggest that rye chromosomes would be preferentially transmitted to the progeny and that an elimination mechanism would act on chromosomes of Thinopyrum and wheat D genome.

Disomic chromosome 3R(3B) substitution causes a complex of meiotic abnormalities in bread wheat Triticum aestivum L.

Triticum aestivum L. lines introgressed with alien chromosomes create a new genetic background that changes the gene expression of both wheat and donor chromosomes. The genes involved in meiosis regulation are localized on wheat chromosome 3B. The purpose of the present study was to investigate the effect of wheat chromosome 3B substituted with homoeologous rye chromosome 3R on meiosis regulation in disomically substituted wheat line 3R(3B). Employing immunostaining with antibodies against microtubule protein, α-tubulin, and the centromere-specific histone (CENH3), as well as FISH, we analyzed microtubule cytoskeleton dynamics and wheat and rye 3R chromosomes behavior in 3R(3B) (Triticum aestivum L. variety Saratovskaya 29 × Secale cereale L. variety Onokhoiskaya) meiosis. The results revealed a set of abnormalities in the microtubule dynamics and chromosome behavior in both first and second divisions. A feature of metaphase I in 3R(3B) was a decrease in the chiasmata number compared with variety Saratovskaya 29, 34.9 ± 0.62 and 41.92 ± 0.38, respectively. Rye homologs 3R in 13.18 % of meiocytes did not form bivalents. Chromosomes were characterized by varying degrees of compaction; 53.33 ± 14.62 cells lacked a metaphase plate. Disturbances were found in microtubule nucleation at the bivalent kinetochores and in their convergence at the spindle division poles. An important feature of meiosis was the asynchronous chromosome behavior in the second division and dyads at the telophase II in 8-13 % of meiocytes, depending on the anther studied. Considering the 3R(3B) meiotic phenotype, chromosome 3B contains the genes involved in the regulation of meiotic division, and substituting 3B3B chromosomes with rye 3R3R does not compensate for their absence.

Meiotic chromosome behaviour of a newly recorded ant-like spider, Myrmarachne melanocephala MacLeay, 1839 from Manipur, India

Cytological and chromosomal studies of ant-like spiders Myrmarachne melanocephala MacLeay, 1839, were undertaken with 12 males captured alive in the months of January to July 2023 from three habitats. The haploid count of male specimens was observed to be 13: 4 acrocentric (including X chromosome), 6 subtelocentric, 2 submetacentric, and 1 metacentric and showed X0 sex determination mechanisms, so the diploid count of the species was 25 (13+12). The single sex chromosome occupied roughly 4-6 per cent of the nuclear volume prominently in the Interphase – Prophase I stage. The structure of X chromosome in interphase stage was circular-rectangular block. The peculiar shape of rod-shaped X chromosome was maintained from early pre-leptotene stage till the end of the division particularly the Prophase I. The synapsis started early from late leptotene and duration of zygotene was long enough to visualize the perfect ones in late zygotene. There were nine diplotene bivalents with interstitial chiasma.

The chromatin-associated 53BP1 ortholog, HSR-9, regulates recombinational repair and X chromosome segregation in the Caenorhabditis elegans germ line.

53BP1 plays a crucial role in regulating DNA damage repair pathway choice and checkpoint signaling in somatic cells; however, its role in meiosis has remained enigmatic. In this study, we demonstrate that the Caenorhabditis elegans ortholog of 53BP1, HSR-9, associates with chromatin in both proliferating and meiotic germ cells. Notably, HSR-9 is enriched on the X chromosome pair in pachytene oogenic germ cells. HSR-9 is also present at kinetochores during both mitotic and meiotic divisions but does not appear to be essential for monitoring microtubule-kinetochore attachments or tension. Using cytological markers of different steps in recombinational repair, we found that HSR-9 influences the processing of a subset of meiotic double-stranded breaks into COSA-1-marked crossovers. Additionally, HSR-9 plays a role in meiotic X chromosome segregation under conditions where X chromosomes fail to pair, synapse, and recombine. Together, these results highlight that chromatin-associated HSR-9 has both conserved and unique functions in the regulation of meiotic chromosome behavior.

Paramutation at the maize pl1 locus is associated with RdDM activity at distal tandem repeats.

Exceptions to Mendelian inheritance often highlight novel chromosomal behaviors. The maize Pl1-Rhoades allele conferring plant pigmentation can display inheritance patterns deviating from Mendelian expectations in a behavior known as paramutation. However, the chromosome features mediating such exceptions remain unknown. Here we show that small RNA production reflecting RNA polymerase IV function within a distal downstream set of five tandem repeats is coincident with meiotically-heritable repression of the Pl1-Rhoades transcription unit. A related pl1 haplotype with three, but not one with two, repeat units also displays the trans-homolog silencing typifying paramutations. 4C interactions, CHD3a-dependent small RNA profiles, nuclease sensitivity, and polyadenylated RNA levels highlight a repeat subregion having regulatory potential. Our comparative and mutant analyses show that transcriptional repression of Pl1-Rhoades correlates with 24-nucleotide RNA production and cytosine methylation at this subregion indicating the action of a specific DNA-dependent RNA polymerase complex. These findings support a working model in which pl1 paramutation depends on trans-chromosomal RNA-directed DNA methylation operating at a discrete cis-linked and copy-number-dependent transcriptional regulatory element.

Meiotic chromosomal behaviour of Artemisia amygdalina Decne: A critically endangered medicinal plant, endemic to the North-western Himalaya

The limited geographic distribution of endemic species renders them more susceptible to extinction risks stemming from a myriad of factors. One such factor could be meiotic constraints, which may directly or indirectly impact the reproductive success of the endemic plant species. Thus, comprehensive research on meiotic behaviour is essential for implementing species conservation measures. This study represents the first report on the male meiotic behaviour of Artemisia amygdalina Decne., a critically endangered endemic medicinal plant species of the Kashmir Himalaya. We procured the plant material from three different locations in the Kashmir Himalaya, and the chromosome number of the species from all sites was found to be 2n=18 (x = 9, n = 9), indicating its diploid nature. Various types of chromosomal abnormalities were found in these populations, including stickiness, univalents, out-of-plate bivalents, laggards, bridges, disturbed telophase, micronuclei, abnormal pollen grains formation and seed set formation. The overall percentage of chromosomal abnormalities varied among the populations, from 8.58 % to 25.16 %. Pollen sterility also varied among different populations, with maximum pollen sterility of 23.21 % and a minimum of 5.96 % across populations. The present findings will not only help in updating the chromosome atlas with the addition of a new report on chromosome number but also contribute to the understanding of the reproductive success of A. amygdalina. Further ecological and reproductive studies can facilitate the creation of effective management and conservation strategies for this species.