Polyploid Plants Research Articles

Autotetraploidization Gives Rise to Differential Gene Expression in Response to Saline Stress in Rice.

Plant polyploidization represents an effective means for plants to perpetuate their adaptive advantage in the face of environmental variation. Numerous studies have identified differential responsiveness to environmental cues between polyploids and their related diploids, and polyploids might better adapt to changing environments. However, the mechanism that underlies polyploidization contribution during abiotic stress remains hitherto obscure and needs more comprehensive assessment. In this study, we profile morphological and physiological characteristics, and genome-wide gene expression between an autotetraploid rice and its diploid donor plant following saline stress. The results show that the autotetraploid rice is more tolerant to saline stress than its diploid precursor. The physiological characteristics were rapidly responsive to saline stress in the first 24 h, during which the elevations in sodium ion, superoxide dismutase, peroxidase, and 1-aminocyclopropane-1-carboxylic acid were all significantly higher in the autotetraploid than in the diploid rice. Meanwhile, the genome-wide gene expression analysis revealed that the genes related to ionic transport, peroxidase activity, and phytohormone metabolism were differentially expressed in a significant manner between the autotetraploid and the diploid rice in response to saline stress. These findings support the hypothesis that diverse mechanisms exist between the autotetraploid rice and its diploid donor plant in response to saline stress, providing vital information for improving our understanding on the enhanced performance of polyploid plants in response to salt stress.

Perspective for genomic-enabled prediction against black sigatoka disease and drought stress in polyploid species.

Genomic selection (GS) in plant breeding is explored as a promising tool to solve the problems related to the biotic and abiotic threats. Polyploid plants like bananas (Musa spp.) face the problem of drought and black sigatoka disease (BSD) that restrict their production. The conventional plant breeding is experiencing difficulties, particularly phenotyping costs and long generation interval. To overcome these difficulties, GS in plant breeding is explored as an alternative with a great potential for reducing costs and time in selection process. So far, GS does not have the same success in polyploid plants as with diploid plants because of the complexity of their genome. In this review, we present the main constraints to the application of GS in polyploid plants and the prospects for overcoming these constraints. Particular emphasis is placed on breeding for BSD and drought-two major threats to banana production-used in this review as a model of polyploid plant. It emerges that the difficulty in obtaining markers of good quality in polyploids is the first challenge of GS on polyploid plants, because the main tools used were developed for diploid species. In addition to that, there is a big challenge of mastering genetic interactions such as dominance and epistasis effects as well as the genotype by environment interaction, which are very common in polyploid plants. To get around these challenges, we have presented bioinformatics tools, as well as artificial intelligence approaches, including machine learning. Furthermore, a scheme for applying GS to banana for BSD and drought has been proposed. This review is of paramount impact for breeding programs that seek to reduce the selection cycle of polyploids despite the complexity of their genome.

Differences in Leaf Morphology and Related Gene Expression between Diploid and Tetraploid Birch (Betula pendula).

Plant polyploidization changes its leaf morphology and leaf development patterns. Understanding changes in leaf morphology and development patterns is a prerequisite and key to studying leaf development in polyploid plants. In this study, we quantified and analyzed the differences in leaf morphology, leaf growth polarity, and leaf size between diploid and tetraploid birches (Betula pendula subsp. pendula), and preliminarily investigated genes involved in leaf growth and development in birch. The results showed significant changes in leaf morphology in tetraploid birches, especially the basal part of the leaf. In addition, the proximal growth rate of tetraploid leaves was altered. The changed proximal growth rate did not affect the growth polarity pattern of tetraploid leaves. The leaf area of tetraploid was significantly larger than that of diploid birch. The difference in leaf size was mainly due to differences in their growth rates in the middle and late stages of leaf development. Increased cell expansion capacity was the major reason for the enormous leaves of tetraploid birch; however, cell proliferation did not contribute to the larger tetraploid leaf. The gene expression of ATHB12 was associated with cell size and leaf area, and may be a critical gene affecting the leaf size in diploid and tetraploid birches. The results will provide valuable insights into plant polyploid leaf development and a theoretical basis for later investigations into the molecular mechanisms underlying the gigantism of tetraploid birch leaves.

AtGCS promoter-driven clustered regularly interspaced short palindromic repeats/Cas9 highly efficiently generates homozygous/biallelic mutations in the transformed roots by Agrobacterium rhizogenes-mediated transformation.

Agrobacterium rhizogenes-mediated (ARM) transformation is an efficient and powerful tool to generate transgenic roots to study root-related biology. For loss-of-function studies, transgenic-root-induced indel mutations by CRISPR/Cas9 only with homozygous/biallelic mutagenesis can exhibit mutant phenotype(s) (excluding recessive traits). However, a low frequency of homozygous mutants was produced by a constitutive promoter to drive Cas9 expression. Here, we identified a highly efficient Arabidopsis thaliana gamma-glutamylcysteine synthetase promoter, termed AtGCSpro, with strong activity in the region where the root meristem will initiate and in the whole roots in broad eudicots species. AtGCSpro achieved higher homozygous/biallelic mutation efficiency than the most widely used CaMV 35S promoter in driving Cas9 expression in soybean, Lotus japonicus, and tomato roots. Using the pAtGCSpro-Cas9 system, the average homozygous/biallelic mutation frequency is 1.7-fold and 8.3-fold higher than the p2 × 35Spro-Cas9 system for single and two target site(s) in the genome, respectively. Our results demonstrate the advantage of the pAtGCSpro-Cas9 system used in ARM transformation, especially its great potential in diploids with multiple-copy genes targeted mutations and polyploid plants with multiplex genome editing. AtGCSpro is conservatively active in various eudicots species, suggesting that AtGCSpro might be applied in a wide range of dicots species.

Triploid cultivars of Cymbidium act as a bridge in the formation of polyploid plants.

Triploid is considered a reproductive barrier and also a bridge in the formation of polyploids. However, few reports are available in Cymbidium. In this study, diploid 'Xiaofeng', sexual triploid 'Yuchan' and 'Huanghe' of Cymbidium were used to evaluate hybridization compatibility of the triploids. Results showed that the sexual triploids were fertile whether they were used as male or female parents. 'Yuchan' produced male gametes of 1x, 1x~2x, 2x, 2x~3x, and 3x at frequencies of 8.89%, 77.78%, 6.67%, 3.33%, and 3.33%, respectively; while 'Huanghe' produced 3.33% 1x, 80.00% 1x~2x, 8.89% 2x, 5.56% 2x~3x, and 2.22% 3x male gametes. The cross of 'Xiaofeng' with 'Yuchan' produced progenies with a wide range of ploidy levels, including one diploid, 34 2×~3× aneuploids, 12 triploids, and one tetraploid, indicating that male gametes produced by sexual triploid were fertile and could be transmitted and fused with egg cells. On the other hand, 10 progenies obtained from the cross of 'Yuchan' × 'Xiaofeng' were all aneuploids. The cross of 'Yuchan' with 'Huanghe' produced 40 progenies including three 2×~3× aneuploids, nine 3×~4× aneuploids, 21 tetraploids, six 4×~5× aneuploids, and one pentaploid, suggesting that 2x gametes, instead of the unreduced ones played a more important role in the formation of tetraploids. The survival rates of the hybrids were all above 80.00%, with the tetraploids at 96.67%. Cytological analysis revealed that during meiosis of sexual polyploids, two chromosome sets of the 2n gamete were inclined to enter into the same daughter cell, resulting in the production of 2x gametes. Our results indicate that the triploid cymbidiums are not reproductive barrier but serve as a bridge in the formation of polyploid plants.

Inconsistent expression of the gigas effect in polyploid Oxalis.

It is well-known that whole genome duplication (WGD) has played a significant role in the evolution of plants. The best-known phenotypic effect of WGD is the gigas effect, or the enlargement of polyploid plant traits. WGD is often linked with increased weediness, which could be a result of fitness advantages conferred by the gigas effect. As a result, the gigas effect could potentially explain polyploid persistence and abundance. We test whether a gigas effect is present in the polyploid-rich geophyte Oxalis, at both organ and cellular scales. We measured traits in conspecific diploid and polyploid accessions of 24 species across the genus. In addition, we measured the same and additional traits in 20 populations of the weedy and highly ploidy-variable species Oxalis purpurea L., including measures of clonality and selfing as a proxy for weediness. Ploidy level was determined using flow cytometry. We found substantial variation and no consistent ploidy-related size difference, both between and within species, and across traits. Oxalis purpurea polyploids did, however, produce significantly more underground biomass and more bulbils than diploids, consistent with a potential role of WGD in the weediness of this species. Our results suggest a more nuanced role for the gigas effect, at least in Oxalis. It may be temporary, short-lived, and inconsistently expressed and retained on evolutionary time scales, but in the short term can contribute to lineage success via increased vegetative reproduction.

Genome evolution: On the nature of trade-offs with polyploidy and endopolyploidy

A new study uncovers a novel trade-off in polyploid plants. While the larger cells of polyploids benefit from increased cell storage and water retention, this comes at the cost of reduced structural stability, potentially impacting survival.

Reactions of Some Grapevine Rootstock Cuttings to Mutagenic Applications

Grapevine (Vitis spp.) is one of the most important socio-economically important plants in the global scale, and the need for its breeding is increasing. In viticulture, mutation is more promising than crossbreeding in breeding new genotypes from natural genetic diversity. Polyploid plants outperform their diploid relatives in several respects. In this study, the efficacy of oryzalin and N2O mutagens in the induction of polyploidy was investigated by applying different doses and durations to the forced cuttings of 41B [Chasselas (Vitis vinifera L.) × Vitis berlandieri Planch)] and Fercal [(Vitis vinifera x Vitis berlandieri) × 333 EM] rootstocks. LD50 values of mutagen applications were determined, morphological and cytological effects were examined by macroscopic, microscopic, and cytological methods. Application time and dose increase of mutagens decreased LD50 values. As a result of mutagen applications, leaf thickness and chlorophyll content of the surviving plants increased. Applications increased stomatal sizes, decreased their density, increased chloroplast numbers, increased leaf thicknesses and partially SPAD values. It was determined that they were not polyploid in the confirmation test performed with flow cytometry (FC) analyses in 4 Fercal and 1 41B samples that were assumed to be mutant by stoma and chloroplast examinations. After that, it was thought that it would be appropriate to try vegetative material with actively dividing cells, such as nodal cuttings, in the studies of obtaining polyploid individuals on grapevine rootstocks

Allopolyploid origin and niche expansion of Rhodiola integrifolia (Crassulaceae)

Polyploidy after hybridization between species can lead to immediate post-zygotic isolation, causing saltatory origin of new species. Although the incidence of polyploidization in plants is high, it is thought that a new polyploid lineage can succeed only if it establishes a new ecological niche divergent from its progenitor lineages. We tested the hypothesis that Rhodiola integrifolia from North America is an allopolyploid produced by R. rhodantha and R. rosea and determined whether its survival can be explained by the niche divergence hypothesis. To this end, we sequenced two low-copy nuclear genes (ncpGS and rpb2) in a phylogenetic analysis of 42 Rhodiola species and tested for niche equivalency and similarity using Schoener’s D as the index of niche overlap. Our phylogeny-based approach showed that R. integrifolia possesses alleles from both R. rhodantha and R. rosea. Dating analysis showed that the hybridization event that led to R. integrifolia occurred ca. 1.67 Mya and niche modeling analysis showed that at this time, both R. rosea and R. rhodantha may have been present in Beringia, providing the opportunity for the hybridization event. We also found that the niche of R. integrifolia differs from that of its progenitors in both niche breadth and optimum. Taken together, these results confirm the hybrid origin of R. integrifolia and support the niche divergence hypothesis for this tetraploid species. Our results underscore the fact that lineages with no current overlapping distribution could produce hybrid descendants in the past, when climate oscillations made their distributions overlap.

Two pathways of 2n gamete formation and differences in the frequencies of 2n gametes between wild species and interspecific hybrids

Epidendrum produces 2n gametes with high frequency. This paper is the first to report on multiple pathways for forming 2n gametes, meiotic defeats, and pre-meiotic chromosome doubling. Unreduced 2n reproductive cells are predominantly involved in pathways that lead to polyploid plants. Although one of the most common pathways for inducing 2n gametes is through meiotic defects, a small set of isolated species alternatively generates 2n gametes from tetraploid pollen mother cells in the pre-meiotic phase. Hence, determining the mechanisms underlying 2n gamete formation is critical to improving breeding programmes and understanding plant evolution. We investigated sporads to reveal the pathway(s) accounting for the formation and frequencies of 2n gametes in wild species and interspecific hybrids in the genus Epidendrum. We investigated different types of sporads with varying frequencies, sizes, and viability in the wild species and hybrids of the genus Epidendrum. Large tetrad-estimated pre-meiotic chromosome doubling was observed in wild species. The Epidendrum is unique in that it forms 2n pollens via two pathways, namely, meiotic defects and pre-meiotic chromosome doubling. These two pathways of 2n pollen formation could influence the high diversity generation of polyploidy with different degrees of heterozygosity and genetic backgrounds in the genus Epidendrum. Therefore, these findings are proposed to influence polyploid breeding of Epidendrum via 2n pollen, helping us understand evolution and speciation via unreduced 2n gamete formation in Orchidaceae.

Induced polyploidy deeply influences reproductive life cycles, related phytochemical features, and phytohormonal activities in blackberry species.

In some cases, polyploidy is an important phenomenon in the evolution of fruit crops. Polyploidy can be used in fruit breeding programs to develop varieties with higher yields and better fruit quality, as well as better adaptation to adverse environmental conditions. In this study, three wild species of blackberry were subjected to different degrees of induced polyploidy, and the effects of which were evaluated on morphological, physiological, and phytohormonal traits. With the aim of gaining a deep insight into the generative phase of plant growth and development, different levels of induced polyploidy were evaluated on the three blackberry species, i.e., Rubus persicus Bioss. (2x, 4x, and 8x), R. caesius L. (2x and 4x), and R. hirtus Schreb. (2x and 4x). The results showed that the polyploid plants performed significantly better than their diploid counterparts in terms of morphological traits such as flower count per spike and berry weight, as well as biochemical traits such as total soluble solids in the leaves. Induced polyploidy increased berry weight and drupe count per fruit. Microscopic examinations revealed a smaller number of viable pollen in the polyploids, compared to the diploids. Electron microscopy showed that the octaploid R. persicus had larger conical cells on the flower surface, compared to the diploid R. persicus. Correlation analysis showed that the ratio of indoleacetic acid to jasmonic acid changed synergistically with the total soluble solids in the leaves during the fruit set. The ploidy level correlated significantly with the number of pistils, leaf green index, total soluble solids in the leaves, and glucose content in floral nectar. Overall, induced polyploidy allowed Rubus to develop advantageous traits that can benefit future breeding programs and expand reproductive research in blackberries.

Organellar transcripts dominate the cellular mRNA pool across plants of varying ploidy levels

Mitochondrial and plastid functions depend on coordinated expression of proteins encoded by genomic compartments that have radical differences in copy number of organellar and nuclear genomes. In polyploids, doubling of the nuclear genome may add challenges to maintaining balanced expression of proteins involved in cytonuclear interactions. Here, we use ribo-depleted RNA sequencing (RNA-seq) to analyze transcript abundance for nuclear and organellar genomes in leaf tissue from four different polyploid angiosperms and their close diploid relatives. We find that even though plastid genomes contain <1% of the number of genes in the nuclear genome, they generate the majority (69.9 to 82.3%) of messenger RNA (mRNA) transcripts in the cell. Mitochondrial genes are responsible for a much smaller percentage (1.3 to 3.7%) of the leaf mRNA pool but still produce much higher transcript abundances per gene compared to nuclear genome. Nuclear genes encoding proteins that functionally interact with mitochondrial or plastid gene products exhibit mRNA expression levels that are consistently more than 10-fold lower than their organellar counterparts, indicating an extreme cytonuclear imbalance at the RNA level despite the predominance of equimolar interactions at the protein level. Nevertheless, interacting nuclear and organellar genes show strongly correlated transcript abundances across functional categories, suggesting that the observed mRNA stoichiometric imbalance does not preclude coordination of cytonuclear expression. Finally, we show that nuclear genome doubling does not alter the cytonuclear expression ratios observed in diploid relatives in consistent or systematic ways, indicating that successful polyploid plants are able to compensate for cytonuclear perturbations associated with nuclear genome doubling.

Induction and identification of tetraploids of pear plants (Pyrus bretschneideri and Pyrus betulaefolia)

Polyploid plants usually exhibit broader leaves, thicker stems, bigger flowers and fruits, dwarfing stature, as well as improved biotic and abiotic resistance. Therefore, increasing the polyploidy is one of the most important strategies used in plant breeding. Here, we reported the successful induction of tetraploids of two pear varieties, ‘duli’ pear (Pyrus betulaefolia), a wildtype pear used for rootstocks in grafting, and ‘Xinli No.7′ pear (Pyrus bretschneideri), a popular cultivated pear variety in China. This was achieved by treating their seeds, shoot tips of tissue cultured or field grown seedlings with colchicine and pendimethalin using three different methods: impregnation, mixed culture and smearing. The best tetraploids induction condition for ‘duli’ pear seeds was impregnating them with 0.4% colchicine for 24 h where the mutation rate of 2.0% was achieved with no chimera found. For shoot tips of tissue cultured seedlings the best condition was impregnating them with 0.2% colchicine where 6.67% of ‘duli’ (48 h) and 13.0% of ‘Xinli No.7′ (24 h) were mutated. In addition, we found that in the mixed culture method the best induction condition for ‘duli’ was mixing with 1.0% colchicine while for ‘Xinli No.7′ it was 0.5% colchicine. In the smear method, for the shoot tips of field grown seedlings of ‘duli’, the best induction condition was treating them with 0.4% colchicine for 24 h. Subsequently, the DNA content of these putative polyploid seedlings was estimated by the flow cytometry, confirming their ploidy nature. Also, we compared the morphological differences between the tetraploid seedlings and the diploid seedlings. Obviously, the tetraploid seedlings appeared to be dwarfed with shorter internode length and reduced internode number of their stems.

Impact of Artificial Polyploidization in Ajuga reptans on Content of Selected Biologically Active Glycosides and Phytoecdysone

Polyploidization in plants, which involves doubling or further multiplying of genome, has the potential to improve the constituents that make medicinal plants, like Ajuga reptans, attractive to the pharmaceutical, cosmetics, and food production industries; botanical pesticide effects could also be derived. The aim of this study was to determine how artificial polyploidization in A. reptans plants affected the composition and quantity of biologically active substances from the glycoside and phytoecdysone families. Diploids and artificial tetraploids of A. reptans were analyzed. Changes in the contents of trans-teupolioside, trans-verbascoside, and 20-hydroxyecdysone were evident in the aboveground parts of the cultivated plants (e.g., leaves and flowers). The tetraploid lines of Ajuga plants displayed variability in, and increased levels of, trans-teupolioside and trans-verbascoside content. The 20-hydroxecdysone content was slightly higher in tetraploids. These findings indicated that Ajuga tetraploids could be used in breeding programs to enhance the yield of substances with potential medicinal and industrial applications.

Evolution of Chromosome Number in Wild Onions (Allium, Amaryllidaceae)

Abstract— Polyploidy has been shown to be a significant driver of diversification among land plants. In addition to whole-genome duplication, other common mechanisms of chromosome number evolution include increases by a multiple of 1.5 in chromosome number due to the fusion of gametes with different ploidy levels (demi-polyploidy), gains or losses of single chromosomes that alter the DNA content of an organism (aneuploidy), or chromosome fission or fusion (ascending dysploidy or descending dysploidy, respectively). Considering the high variability in chromosome number transitions across multiple clades within angiosperms and the ancient genome duplication events responsible for their diversity, more studies of large polyploid systems are necessary to close the gaps in understanding chromosomal evolution in polyploid plants. Allium L. (Amaryllidaceae) is an ideal candidate for polyploid research because it is a large clade that includes numerous natural populations of diploid and polyploid species. Species of Allium mainly occupy temperate climates in the Northern Hemisphere and include economically important ornamentals and cultivated crops such as leeks, garlic, chives, and onions. Here, we used a molecular phylogeny of Allium to examine chromosomal evolution with chromEvol v. 2.0 which uses likelihood-based methods for inferring the pattern of chromosome number change across a phylogeny. The best-fit model of chromosomal evolution indicated that chromosome transitions within Allium occurred through the constant gains and losses of single chromosomes as well as demi-polyploidization events, with the rate of chromosome gain events being approximately 2.5 to 4.5 times more likely to occur than demi-polyploidization and loss events, respectively.

Induction of 2n pollen by colchicine during microsporogenesis to produce polyploids in herbaceous peony (Paeonia lactiflora Pall.)

Polyploid breeding is an effective way to improve ornamental traits of horticultural plants. Colchicine, an anti-mitotic agent, is widely used to induce 2n gametes in horticultural plants, allowing for the production of polyploid plants through sexual hybridization. Based on an understanding of flower bud development and meiosis in pollen mother cells (PMCs) of Paeonia lactiflora cv. ‘Fen Yu Nu’, 2n pollen were obtained by injecting different concentrations of colchicine (0.1, 0.2, 0.3, 0.4 or 0.5%) into flower buds multiple times (once, twice or three times with a 24-h interval in between). External morphology, viability and possible hybridization of colchicine-induced 2n pollen were observed. Finally, P. lactiflora cv. ‘Lian Tai’ was pollinated with colchicine-induced pollen from ‘Fen Yu Nu’. Polyploids were identified among offspring by flow cytometry, chromosome counting, and molecular assessment using simple sequence repeats (SSR). Flower bud diameter of herbaceous peony was related with meiosis in PMCs. The most suitable period to induce 2n pollen was when most PMCs were in prophase I of meiosis, and when flower buds were about 11-12 mm in diameter. The highest induced incidence (%) of 2n pollen was 47.39% following treatment with 0.4% colchicine solution injected twice. The average diameter of 2n pollen was 46.01 μm, which was 1.53-fold wider than haploid pollen. SEM results indicated that 2n pollen were considerably different from haploid pollen in terms of external shape and surface decoration, and 2n pollen were mostly spherical or with an irregular shape. Induced 2n pollen could germinate in vitro and on the stigma of the female parent, but abnormalities such as pollen tube bifurcation or thickening were observed in both germinated states. The percentage of germinated 2n pollen in vitro was 12.78%. In addition, two tetraploids (2n = 4x = 20) were obtained by 2n pollen hybridization. SSR analysis indicated that tetraploids were produced by the spontaneous fusion of 2n female gametes with colchicine-induced 2n male gametes. This indicates that colchicine did not entirely inactivate 2n pollen. The production of few polyploids may be due to the abnormal germination of 2n pollen and obstacles in ploidy hybridization. Our findings fortify the foundation for Paeonia polyploid breeding.

Exploiting Unreduced Gametes for Improving Ornamental Plants.

The formation of gametes with somatic chromosome number or unreduced gametes (2n gametes) is an important process involved in the origin of polyploid plants in nature. Unreduced gametes are the result of meiotic mutations occurring during micro- and mega-sporogenesis. 2n gametes have been identified or artificially induced in a large number of plant species. Breeding of plants through 2n gametes can be advantageous because it combines genetic effects of polyploidy with meiotic recombination and sexual hybridization to produce tremendous genetic variation and heterosis. 2n gametes also occur in ornamental plants, but the potential of using 2n gametes in ornamental plant breeding has not been extensively exploited. Ornamental plants are primarily produced for their esthetic appearance and novelty, not for food and yield, and they can be readily propagated through vegetative means. Triploids, tetraploids, and plants with even higher ploidy levels produced through 2n gametes can be propagated through tissue culture to fix their phenotypes, thus leading to the development of new cultivars. In this review article, we intend to discuss the mechanisms underlying the formation of 2n gametes, techniques for 2n gamete identification, methods for enhancing 2n gamete formation, and the current status in the use of 2n gametes for development of novel ornamental plants. We believe that polyploidy breeding through 2n gametes represents a viable way of developing new cultivars, new species, and even new genera of ornamental plants.

The nature and genomic landscape of repetitive DNA classes in Chrysanthemum nankingense shows recent genomic changes.

Tandemly repeated DNA and transposable elements represent most of the DNA in higher plant genomes. High-throughput sequencing allows a survey of the DNA in a genome, but whole-genome assembly can miss a substantial fraction of highly repeated sequence motifs. Chrysanthemum nankingense (2n = 2x = 18; genome size = 3.07 Gb; Asteraceae), a diploid reference for the many auto- and allopolyploids in the genus, was considered as an ancestral species and serves as an ornamental plant and high-value food. We aimed to characterize the major repetitive DNA motifs, understand their structure and identify key features that are shaped by genome and sequence evolution. Graph-based clustering with RepeatExplorer was used to identify and classify repetitive motifs in 2.14 millions of 250-bp paired-end Illumina reads from total genomic DNA of C. nankingense. Independently, the frequency of all canonical motifs k-bases long was counted in the raw read data and abundant k-mers (16, 21, 32, 64 and 128) were extracted and assembled to generate longer contigs for repetitive motif identification. For comparison, long terminal repeat retrotransposons were checked in the published C. nankingense reference genome. Fluorescent in situ hybridization was performed to show the chromosomal distribution of the main types of repetitive motifs. Apart from rDNA (0.86 % of the total genome), a few microsatellites (0.16 %), and telomeric sequences, no highly abundant tandem repeats were identified. There were many transposable elements: 40 % of the genome had sequences with recognizable domains related to transposable elements. Long terminal repeat retrotransposons showed widespread distribution over chromosomes, although different sequence families had characteristic features such as abundance at or exclusion from centromeric or subtelomeric regions. Another group of very abundant repetitive motifs, including those most identified as low-complexity sequences (9.07 %) in the genome, showed no similarity to known sequence motifs or tandemly repeated elements. The Chrysanthemum genome has an unusual structure with a very low proportion of tandemly repeated sequences (~1.02 %) in the genome, and a high proportion of low-complexity sequences, most likely degenerated remains of transposable elements. Identifying the presence, nature and genomic organization of major genome fractions enables inference of the evolutionary history of sequences, including degeneration and loss, critical to understanding biodiversity and diversification processes in the genomes of diploid and polyploid Chrysanthemum, Asteraceae and plants more widely.

Alkaline Stress Induces Different Physiological, Hormonal and Gene Expression Responses in Diploid and Autotetraploid Rice.

Saline−alkaline stress is a critical abiotic stress that negatively affects plants’ growth and development. Considerably higher enhancements in plant tolerance to saline−alkaline stress have often been observed in polyploid plants compared to their diploid relatives, the underlying mechanism of which remains elusive. In this study, we explored the variations in morphological and physiological characteristics, phytohormones, and genome-wide gene expression between an autotetraploid rice and its diploid relative in response to alkaline stress. It was observed that the polyploidization in the autotetraploid rice imparted a higher level of alkaline tolerance than in its diploid relative. An eclectic array of physiological parameters commonly used for abiotic stress, such as proline, soluble sugars, and malondialdehyde, together with the activities of some selected antioxidant enzymes, was analyzed at five time points in the first 24 h following the alkaline stress treatment between the diploid and autotetraploid rice. Phytohormones, such as abscisic acid and indole-3-acetic acid were also comparatively evaluated between the two types of rice with different ploidy levels under alkaline stress. Transcriptomic analysis revealed that gene expression patterns were altered in accordance with the variations in the cellular levels of phytohormones between diploid and autotetraploid plants upon alkaline stress. In particular, the expression of genes related to peroxide and transcription factors was substantially upregulated in autotetraploid plants compared to diploid plants in response to the alkaline stress treatment. In essence, diploid and autotetraploid rice plants exhibited differential gene expression patterns in response to the alkaline stress, which may shed more light on the mechanism underpinning the ameliorated plant tolerance to alkaline stress following genome duplication.

Heat shock-induced failure of meiosis I to meiosis II transition leads to 2n pollen formation in a woody plant.

The formation of diploid gametes through chromosome doubling is a major mechanism of polyploidization, diversification, and speciation in plants. Unfavorable climate conditions can induce or stimulate the production of diploid gametes during meiosis. Here, we demonstrated that heat shock stress (38°C for 3 or 6 h) induced 2n pollen formation, and we generated 42 triploids derived from heat shock-induced 2n pollen of Populus canescens. Meiotic analysis of treated pollen mother cells revealed that induced 2n pollen originated from the complete loss of meiosis II (MII). Among the 42 triploids, 38 triploids derived from second division restitution (SDR)-type 2n pollen and 4 triploids derived from first division restitution-type 2n pollen were verified using simple sequence repeats(SSR) molecular markers. Twenty-two differentially expressed genes related to the cell cycle were identified and characterized by expression profile analysis. Among them was POPTR_0002s08020g (PtCYCA1;2), which encodes a type A Cyclin CYCA1;2 that is required for the meiosis I (MI) to MII transition. After male flower buds were exposed to heat shock, a significant reduction was detected in PtCYCA1;2 expression. We inferred that the failure of MI-to-MII transitions might be associated with downregulated expression of PtCYCA1;2, leading to the formation of SDR-type 2n pollen. Our findings provide insights into mechanisms of heat shock-induced 2n pollen formation in a woody plant and verify that sensitivity to environmental stress has evolutionary importance in terms of polyploidization.