Increasing Ploidy Level Research Articles

High mountains of central Europe as a refuge of surprising cytotype diversity of Huperzia selago (Lycopodiaceae)

Polyploidization is pivotal in plant speciation, affecting adaptability, ecological tolerance and specific geographical distribution patterns. While cytotype diversity has been extensively studied in angiosperms and ferns, knowledge in homosporous lycophytes remains very limited. Our study addresses this gap, focusing on the homosporous lycophyte Huperzia selago in Central Europe. The genome size of 1330 individuals from 208 populations were assessed via flow cytometry, revealing five distinct cytotypes (2x, 3x, 4x, 5x, 6x). Challenging chromosome counting using gemmae roots yielded average counts of 140 (2x), 204 (3x), and 262 (4x) chromosomes. The hexaploid genome size (29 pg) approached the upper limits reported for lycophytes. Tetraploids were the most abundant (72.7%), while triploids (21.3%) were less common, while the remaining cytotypes comprised less than 5% of the dataset. Geographical separation of cytotypes was not observed. However, uncommon cytotypes were associated with the Alps and triploids also occured in the highest parts of Western Carpathians. Around 27.3% of populations exhibited multiple cytotypes. Except for atypical diploids, spore abortion differed between even (21.8%) and odd ploidies (65.2%). Microcharacter (stoma, spore) proportions did not linearly correspond with increasing ploidy levels. The high ploidy-level diversity and cytotype coexistence in Central European H. selago match the well documented patterns in ferns and angiosperms. These findings provide valuable insights into lycophyte polyploidy, underscoring the necessity for broader geographical sampling and application of molecular studies to elucidate phylogenetic relationships and taxonomic classifications within the genus Huperzia.

Vascular system of rice leaves depending on ploidy level

The current paper has provided data on the study of the vascular system of flag leaves of rice plants grown in a greenhouse in the period 2022–2023. The objects of the study were 24 rice samples developed at the FSBSI “ARC “Donskoy” using the method of androgenesis and cell culture. The purpose of the work was to determine the degree of development of the vascular system of flag leaves of regenerated rice plants with different ploidy levels (1n, 2n, 4n). When studying the vascular system of plant leaves (the number and area of bundles), it was found that haploid plants had smaller vascular bundles compared to diand tetraploid samples. The average beam diameter had dimensions of 59.5, 69.3 and 75.3 µm, the area of one beam was 2815.6, 3827.2 and 4540.5 µm², respectively. In the leaves of rice samples, a larger number of small and large vascular bundles have been formed with increasing ploidy level. Their number was 36–44 in haploids, 40–52in diploids, 52–60 in tetraploids. The average number of bundles was 40.3, 46.6 and 55.2, respectively. The venation pattern was individual for each sample. Between single large bundles there are from one to seven small veins, most often 4–6 pieces. Thus, samples with different ploidy levels differ in the anatomical structure of the leaves, which ultimately affects their morphology and productivity.

RDNA-FISH pattern in selected <i>Hieracium</i> species representing different ploidy levels

Polyploid species of <i>Hieracium</i> s.str. are near-obligatory apomicts capable of producing non-reduced pollen grains and intrageneric crossing with sexual diploid species. Most probably, such a mating system leads to the emergence of separate lineages of apomictic plants with increased ploidy levels, each of which has a complete set of genomes of its putative (most probably triploid) apomictic parent. The correct identification of such lineages could facilitate taxonomic and phylogenetic research in <i>Hieracium</i>. In this paper, we analyzed the distribution of the 5S and 45S rDNA segments in the karyotypes of six selected <i>Hieracium</i> taxa, representing all ploidy levels reported in the genus: <i>H. alpinum</i> 2x, <i>H. alpinum</i> 3x, <i>H. schustleri</i> 4x, <i>H. chrysostyloides</i> 5x (<i>H</i>. sect. <i>Alpina</i>) and <i>H. bifidum</i> 3x, <i>H. levicaule</i> 3x (<i>H</i>. sect. <i>Bifida</i>). The analyzed rDNA markers suggest that two taxa of the higher ploidy level (4x and 5x) belonging to <i>H</i>. sect. <i>Alpina</i> inherited three genomes from the triploid <i>H. alpinum</i>, which has 2:1 genomic composition with two genomes from diploid <i>H. alpinum</i> and one genome with an untypical 5S rDNA-bearing chromosome of unknown origin. <i>H. bifidum</i> and <i>H. levicaule</i> differ from each other in the rDNA distribution pattern, which suggests that <i>H</i>. sect. <i>Bifida</i> may be a less homogenous group of species.

Water stress and increased ploidy level enhance antioxidant enzymes, phytohormones, phytochemicals and polyphenol accumulation of tetraploid induced wallflower

Wallflower (Erysimum cheiri (L.) Crantz) is a profitable plant in the food, cosmetic, and pharmaceutical industries, as it produces valuable phenolic compounds. It is also an important flower in the floriculture industry cultivated mainly in the gardens and pots. Nonetheless, water scarcity as a worldwide phenomenon has deteriorated its growth and productivity. A main mechanism for drought tolerance is genome duplication and increased ploidy level, where more metabolites are produced. In this regard, diploid and tetraploid-induced plants of E. cheiri were subjected to well-watered (100% field capacity) and water deficit (50% field capacity) conditions for 30 days. Under water deficit conditions, the tetraploid wallflowers conserved their relative water content (RWC) at 85%, while the RWC of diploid plants was reduced to 68%. The chromosome-doubled plants showed advantages under water stress due to their higher osmotic adjustment by increased total soluble sugar and proline content, stronger antioxidant enzymes’ activity of SOD, CAT, APX, and POD, and enhanced non-enzymatic defense systems (i.e., total phenolic and flavonoid contents), compared with their control diploids. Also, stress hormones such as abscisic acid, salicylic acid, jasmonic acid, and auxins were increased in tetraploid plants under low water conditions, though cytokinin and gibberellin phytohormones were decreased as an adaptive strategy in the stress condition. Furthermore, chlorogenic acid and gallic acid were identified as the main phenolic compounds in 4x Erysimum leaves. The superiority of tetraploids over the diploid plants under water stress might be attributed to the extensive reprogramming of genes involved in hormonal signaling, reactive oxygen species detoxification, osmotic adjustment, and enhanced antioxidant defense system. The polyploidization technique combined with water scarcity, have proved fruitful for the production of excess secondary metabolites for various industrial uses.

The Abundant and Unique Transcripts and Alternative Splicing of the Artificially Autododecaploid London Plane (Platanus × acerifolia).

Transcription and alternative splicing (AS) are now appreciated in plants, but few studies have examined the effects of changing ploidy on transcription and AS. In this study, we showed that artificially autododecaploid plants of London plane (Platanus × acerifolia (Aiton) Willd) had few flowers relative to their hexaploid progenitors. Transcriptome analysis based on full-length Oxford Nanopore Technologies (ONTs) and next-generation sequencing (NGS) revealed that the increased ploidy level in P. × acerifolia led to more transcribed isoforms, accompanied by an increase in the number of isoforms per gene. The functional enrichment of genes indicated that novel genes transcribed specifically in the dodecaploids may have been highly correlated with the ability to maintain genome stability. The dodecaploids showed a higher number of genes with upregulated differentially expressed genes (DEGs) compared with the hexaploid counterpart. The genome duplication of P. × acerifolia resulted mainly in the DEGs involved in basic biological pathways. It was noted that there was a greater abundance of alternative splicing (AS) events and AS genes in the dodecaploids compared with the hexaploids in P. × acerifolia. In addition, a significant difference between the structure and expression of AS events between the hexaploids and dodecaploids of Platanus was found. Of note, some DEGs and differentially spliced genes (DSGs) related to floral transition and flower development were consistent with the few flower traits in the dodecaploids of P. × acerifolia. Collectively, our findings explored the difference in transcription and AS regulation between the hexaploids and dodecaploids of P. × acerifolia and gained new insight into the molecular mechanisms underlying the few-flower phenotype of P. × acerifolia. These results contribute to uncovering the regulatory role of transcription and AS in polyploids and breeding few-flower germplasms.

Antimicrobial and Hepatoprotective Activity of Different Cytotypes of Taraxacum officinale from the Indian Himalayas

ABSTRACT Three cytotypes (2×), (3×), and (4×) of Taraxacum officinale (Dandelion) from the Lahaul-Spiti region of Himachal Pradesh, India, were examined for their phytochemical composition and antimicrobial and hepatoprotective properties. The diploid cytotype had the highest concentration of cichoric acid (1.20% w/w), followed by the triploid and tetraploid cytotypes (0.21 and 0.14% w/w, respectively). The cichoric acid content decreased with increasing ploidy level. The ethanol extract of leaves from diploid cytotype exhibited the highest antibacterial activities against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia, and Pseudomonas aeruginosa. As shown by the reduction in the activity of marker enzymes (AST, ALT, and ALP) and the restoration of total protein, bilirubin levels, and liver weight, the hepatoprotective activity was the greatest in the diploid cytotype. Although polyploidy is a key factor in plant evolution, it does not always have the same impact on secondary metabolites.

Cultivar-dependent phenotypic and chemotypic responses of drug-type Cannabis sativa L. to polyploidization.

Cannabis sativa L. is a plant with a wide range of potential medicinal applications. In recent years, polyploidy has gained attention as a potential strategy for rapidly improving C. sativa, which, unlike other modern crops, has not yet benefitted from this established biotechnological application. Currently, no reports on high THCA and CBDA drug-type polyploid cultivars have been published. Moreover, it still needs to be clarified if different cultivars react similarly to polyploidization. For these reasons, we set out to evaluate and compare the phenotype and chemotype of three high Δ9-tetrahydrocannabinolic acid (THCA) and one high cannabidiolic acid (CBDA) drug-type cultivars in their diploid, triploid and tetraploid state through agronomic and metabolomic approaches. Our observations on plant morphology revealed a significant increase in plant height and leaf size with increasing ploidy levels in a cultivar-dependent manner. In contrast, cannabinoids were negatively affected by polyploidization, with the concentration of total cannabinoids, THCA, CBDA and cannabigerolic acid (CBGA) decreasing significantly in higher ploidy levels across all four cultivars. Headspace analysis of volatiles revealed that total volatile content decreased in triploids. On the other hand, tetraploids reacted differently depending on the cultivars. Two THCA dominant cultivars showed an increase in concentrations, while in the other two cultivars, concentrations decreased. Additionally, several rare compounds not present in diploids appeared in higher ploidy levels. Moreover, in one high THCA cultivar, a couple of elite tetraploid genotypes for cannabinoid and volatile production were identified, highlighting the role of cultivar and genotypic variability as an important factor in Cannabis sativa L. polyploids. Overall, our observations on plant morphology align with the giga phenotype observed in polyploids of other plant species. The decrease in cannabinoids and volatiles production in triploids have relevant implications regarding their commercial use. On the other hand, this study found that tetraploidization is a suitable approach to improve Cannabis sativa L. medicinal potential, although the response is cultivar and genotype-dependent. This work lays the ground for further improving, evaluating and harnessing Cannabis sativa L. chemical diversity by the breeding, biotechnological and pharmaceutical sectors.

Creation and characteristics of tetraploid and mixoploid centipedegrass

AbstractPloidy manipulation has been used in many crop improvement programs to develop plant species with wider adaptability and desirable traits. The objectives of this study were to create stable tetraploid centipedegrass (Eremochloa ophiuroides [Munro] Hack.) lines and evaluate them for beneficial traits. To generate polyploid lines, “TifBlair” (2n = 2x = 18) seeds were exposed to gamma radiation and callus was generated using tissue culture, exposed to glyphosate and regenerated into plantlets. After 5 years of continuous propagation, two lines, Hongliang Wang (HW)16 and HW123, were found to be tetraploid, and one line, HW61, was found to be a mixoploid using flow cytometry and chromosome counts. Nuclear DNA contents for diploid and tetraploid lines ranged from 1.97 to 2.10 and 4.14 to 4.30 pg 2C−1, respectively. Ten centipedegrass lines were genotyped using seven simple sequence repeat markers and clustered by origin (derived from TifBlair or the University of Georgia breeding program). Six lines were evaluated for morphological and physiological traits under greenhouse conditions. Trait evaluation showed that tetraploid lines had larger stomata and leaf width and reduced stomatal density. For all other traits evaluated under greenhouse conditions, although significant differences were observed between lines, no consistent differences separated diploid and tetraploid lines. Between the tetraploid lines, HW16 had higher evapotranspiration, percentage green cover, and relative water content than HW123, whereas HW123 had a higher average clipping yield and root width than HW16. In conclusion, irradiation and tissue culture are valuable techniques for the generation of stable polyploid lines, and an increased ploidy level led to larger vegetative structures in centipedegrass.

Ploidy effects on the relationship between floral phenotype, reproductive investment, and fitness in an autogamous species complex

AbstractPremiseThe relationships between reproductive investment, phenotype, and fitness have been broadly studied in cross‐pollinated plants in contrast to selfing species, which are considered less interesting in this area because they are supposed to be a dead end in any evolutionary pathway. Still, selfing plants are unique systems to study these questions since the position of reproductive structures and traits related to flower size play an important role in female and male pollination success.MethodsErysimum incanums.l. is a selfing species complex that has three levels of ploidy (diploids, tetraploids, and hexaploids) and traits that are typically associated with the selfing syndrome. Here, we used 1609 plants belonging to these three ploidies to characterize the floral phenotype and spatial configuration of reproductive structures, reproductive investment (pollen and ovule production), and plant fitness. Then, we used structural equation modelling to analyze the relationship between all these variables across ploidy levels.ResultsAn increase in ploidy level leads to bigger flowers with anthers exserted farther and more pollen and ovules. In addition, hexaploid plants had higher absolute values for herkogamy, which is positively correlated with fitness. Ovule production significantly mediated the natural selection acting on different phenotypic traits and pollen production, a pattern that is maintained across ploidies.ConclusionsThe changes in floral phenotypes, reproductive investment, and fitness with ploidy level suggest that genome duplication can be a driver for transitions in reproductive strategy by modifying the investment in pollen and ovules and linking them with plant phenotype and fitness.

Induction of Polyploidy in Citrus Rootstocks through In Vitro Colchicine Treatment of Seed-Derived Explants

Polyploidy, frequently observed in citrus species, aids in achieving better adaptation to environmental stresses. In this context, the current work aims to develop stable tetraploids in citrus rootstock cultivars, viz., Rough lemon, Rangpur lime and Alemow, through in vitro colchicine treatments. Seed-derived explants were obtained by culturing sterile seeds on MS basal media. Seedlings with a size of 5–8 mm (hypocotyl) were exposed to colchicine treatment. After treatment, the surviving seedlings were minigrafted onto six-month-old rootstock for better growth. Colchicine concentrations of 0.1%, 0.2%, 0.3% and control for durations of 16 or 24 h were tested with respect to the induction of polyploidisation. Treatment with 0.1% colchicine for 24 h resulted in high rates of mutation for polyploidisation and showed the highest tetraploid induction percentage (18.3%) in all the rootstock cultivars. High colchicine concentration and long exposure time decreased the survival of the observed seedlings. Flow cytometry and cytological methods were used for confirmation of autotetraploidy in the analysed samples. The surviving seedlings were identified on the basis of morphological and cytological variables, such as leaf area and stomata size, which significantly increased with increasing ploidy level. The proposed method was found to be an effective way to induce the polyploidy in Rangpur lime, Rough lemon and Alemow rootstocks.

Polyploidisation effect on trichome density in interspecific hybrids of Pennisetum glaucum (L.) R. Br. × Pennisetum purpureum (K.) Schum

This study is aimed at testing the efficiency of colchicine on inducing polyploidy in Pennisetum glaucum (L.) R. Br. ? P. purpureum (K.) Schum (BNH) and to investigate the effect of polyploidy on trichome density. Root cuttings of 4 hybrids and two check varieties were treated with different concentrations of colchicine (0, 0.05%, 0.1% and 0.2%) for different time durations (12 and 24 hrs). The ploidy level of plants was confirmed by cytogenetic studies using conventional protocol. Colchicine concentration of 0.1% for 24 hrs was found to induce polyploidy in PBN 233. Trichome density was investigated using Scanning Electron Microscopy at 100X. During the study, it was found that the trichome density decreases with increase in ploidy level.

Phytochemical Diversity in Relation to Cytogenetic Variability in Inula racemosa Hook.f., an Endangered Medicinal Plant of Himalayas.

Inula racemosa, a resourceful critically endangered medicinal herb in the Himalayas is traditionally utilized to cure various human disorders. The species is a wealthy source of sesquiterpene lactones has many pharmacological properties. To quantify and identify the best genetic stocks for a maximum build-up of desired metabolites (isoalantolactone and alantolactone) among existent cytotypes in the species, LC-MS/MS analysis was made. The other comprehensive experiments carried out at present included detailed meiotic examinations of different populations collected from different areas of Kashmir Himalayas. The results presented the occurrence of variable chromosome numbers as n=10 and n=20 in different populations, but the tetraploid cytotype (n=20) is new for the species. The LC-MS/MS investigation revealed significant variability in the content of sesquiterpene lactones in different plant tissues (stem, leaf, and root). An upsurge in the quantity of isoalantolactone and alantolactone was noticed with increasing ploidy levels along the increasing altitudes. Therefore, a habit to accumulate abundant quantities of secondary metabolites and increased adaptability by species/cytotypes thriving at higher altitudes is seen among tetraploid cytotypes during the present investigation. Also, the chromosomal variations seem to enhance the flexibility of polyploid species primarily at upper elevations. Thus, the present study strongly provides quantification of elite cytotypes/chemotypes with optimum concentration of secondary metabolites.

Near-Hexaploid and Near-Tetraploid Aneuploid Progenies Derived from Backcrossing Tetraploid Parents Hibiscus syriacus × (H. syriacus × H. paramutabilis).

Hibiscus syriacus, azalea, is an important woody ornamental shrub planted throughout many temperate and subtropical regions of the world. However, flower size is smaller in this species than some of its relatives. To increase flower size, interspecific hybridization has been used, and such hybrid cultivars are usually characterized by larger flowers, increased vigor, diverse leaf shapes, and reduced fertility. Our earlier studies have shown that these hybrid cultivars could backcross with H. syriacus when used as male parents. To understand the breeding potential of these hybrid cultivars, two popular tetraploid hybrid cultivars, ‘Lohengrin’ and ‘Resi’, were used as pollen parents to backcross several tetraploid H. syriacus cultivars. As a result, 28.76% and 64.4% of ‘Lohengrin’ and ‘Resi’ progenies exhibited larger flowers than both of their parents. Interestingly, 14 of 18 progenies of ‘Resi’ were putative hexaploids, whereas 19 tested ‘Lohengrin’ progenies were tetraploid. Because putative hexaploid progenies were only observed among progenies of ‘Resi’, this hybrid cultivar appears to produce unreduced gametes. In addition, among the 14 putative hexaploids derived from ‘Resi’, 11 had larger flowers than both of their parents and their tetraploid siblings (p < 0.05). The 45S rDNA and 5S rDNA locus segregation among those BC1F1 progenies was tested by fluorescent in situ hybridization (FISH), and the wide range of 45S rDNA signal numbers among siblings indicated that these aneuploids resulted from unequal segregation or chromosome rearrangement. Chromosome counting confirmed aneuploidy among BC1F1 progenies. Ploidy diversity and aneuploidy have been known to contribute to various elements of morphological diversity, such as larger flower size and reduced fertility, which are important in ornamental plant breeding. The present study demonstrated the breeding potential of interspecific Hibiscus cultivars for increasing ploidy level and flower size.

Examination ofwhite blood cell indicators for three different ploidy level sturgeon species reared inanindoor recirculation aquaculture system for one year.

Functional diploid Acipenser ruthenus, functional tetraploid Acipenser gueldenstaedtii and functional hexaploid Acipenser brevirostrum juveniles were sampled monthly for one year, and the white blood cell indicators were determined. The total number ofleukocytes (TL) was 40.93± 17.24× 109/l for the diploids, 20.63± 11.20×109/l for the tetraploids, 14.13± 7.72×109/l for the hexaploids. The TL decreased with anincreasing ploidy level. The highest number ofleukocytes was reached during September and October for A.ruthenus and A.brevirostrum, from October toJanuary for A.gueldenstaedtii (astatistically significant finding). The lymphocytes dominated (76.89-80.14%) inthe differential counts and were found tobe reduced inJune and July ineach group. Granulocytes were represented byneutrophils and eosinophils. Counting from all the leukocytes, the neutrophils represented 13.0-18.7% and eosinophils represented 5.7-6.1%. Increasing number ofnuclear segments inthe granulocytes was dependent onthe increasing ploidy level. Nuclear segmentation inthe lymphocytes was acommon finding inhigher ploidy level groups. The data suggest asignificant effect ofploidy level onthe total number ofleukocytes and morphological nuclear changes inthe granulocytes and lymphocytes. The seasonal variation inthe differential leukocyte counts depends onthe species and the influence ofvarious external conditions rather than the ploidy level.

In vitro amphidiploid induction of a distant hybrid Populus simonii × P. euphratica cv. ‘Xiaohuyang-2’ and its effect on plant morphology and anatomy

Abstract Highly gametic sterility of a distant hybrid Populus simonii × P. euphratica cv. ‘Xiaohuyang-2’ restricts its utilization in breeding programs of Populus. Amphidiploid induction by somatic chromosome doubling is expected to restore its gametic fertility. In this study, nodal-segment and leaf explants of ‘Xiaohuyang-2’ were used to induce chromosome doubling with colchicine in vitro. Although chromosome doubling of the nodal-segment explants only produced mixoploids, the treatments of leaf explants on adventitious bud regeneration medium successfully produced 4 amphidiploids, which might be attributed to the direct organogenesis of the adventitious buds on the leaf explants. This is the first report of amphidiploid induction in a distant hybrid between Populus section Tacamahaca and sect. Turanga. The highest amphidiploid induction frequency was 16.7 %. Both the explant survival rate and polyploidization frequency were significantly affected by colchicine concentration and exposure time. The amphidiploid plants significantly differed from the diploid and mixoploid plants in morphological and anatomical characteristics. They had larger, thicker, and greener leaves than the diploids and mixoploids. The increase in ploidy level also resulted in changes in stomatal features. The induced amphidiploid plants of the distant hybrid ‘Xiaohuyang-2’ are expected to play important roles in breeding programs of Populus in the future, which can be used as a bridge parent with the ability of unreduced gamete formation to cross with fast-growth germplasms to produce triploids pyramiding desirable traits of fast growth, easy cutting propagation, and salt and drought tolerances.

Gradual genome size evolution and polyploidy in Allium from the Qinghai-Tibetan Plateau.

Genome size is an important plant trait, with substantial interspecies variation. The mechanisms and selective pressures underlying genome size evolution are important topics in evolutionary biology. There is considerable diversity in Allium from the Qinghai-Tibetan Plateau, where genome size variation and related evolutionary mechanisms are poorly understood. We reconstructed the Allium phylogeny using DNA sequences from 71 species. We also estimated genome sizes of 62 species, and determined chromosome numbers in 65 species. We examined the phylogenetic signal associated with genome size variation, and tested how well the data fit different evolutionary models. Correlations between genome size variations and seed mass, altitude and 19 bioclimatic factors were determined. Allium genome sizes differed substantially between species and within diploids, triploids, tetraploids, hexaploids and octaploids. Size per monoploid genome (1Cx) tended to decrease with increasing ploidy levels. Allium polyploids tended to grow at a higher altitude than diploids. The phylogenetic tree was divided into three evolutionary branches. The genomes in Clade I were mostly close to the ancestral genome (18.781 pg) while those in Clades II and III tended to expand and contract, respectively. A weak phylogenetic signal was detected for Allium genome size. Furthermore, significant positive correlations were detected between genome size and seed mass, as well as between genome size and altitude. However, genome size was not correlated with 19 bioclimatic variables. Allium genome size shows gradual evolution, followed by subsequent adaptive radiation. The three well-supported Allium clades are consistent with previous studies. The evolutionary patterns in different Allium clades revealed genome contraction, expansion and relative stasis. The Allium species in Clade II may follow adaptive radiation. The genome contraction in Clade III may be due to DNA loss after polyploidization. Allium genome size might be influenced by selective pressure due to the conditions on the Qinghai-Tibetan Plateau (low temperature, high UV irradiation and abundant phosphate in the soil).

Induction of polyploidy through colchicine in cotton (Gossypium herbaceum) and its conformity by cytology and flow cytometry analyses

Cotton is one of the most important fibre crops in the world. An increase in ploidy level was observed in diploid cotton species namely Gossypium herbaceum in the experiment, through colchicine application. There were significant growth variations noticed during the induction of polyploidy in the cotton plants depending upon the concentration of colchicine, duration of the treatments and genotypes taken. An increase in the concentration of colchicine or the duration of the treatment had a retardation effect on seed viability in G. herbaceum. The hypocotyls of length between 4 and 8 mm were found to be most responsive to colchicine treatment. The root meristem of G. herbaceum, treated with 0.2 and 0.4% colchicine exhibited the most number of tetraploid cells during 16 h of treatment. The increase in the concentration of colchicine, along with an increase in duration of treatment led to the chromosomal abnormalities in the wild cotton species. Seed treatment for colchicine application, was the most efficient and reliable method when compared to Petri-plate application and cotton swabbing treatments for inducing polyploidy in diploid cotton.

Induction of Autotetraploidy and its effects on morphophysiological traits in some annual and perennial medics

In order to study of the effects of polyploidy on morph-physiological traits in some annual and perennial medics, five annual and three perennial diploid medics were subjected to different concentrations of colchicine solution (0.1%, 0.5%, 1% and 2%) in mitosis stage. The induced tetraploids were identified by counting stomata guard cells chloroplasts. The highest survival rate and tetraploidy induction with average values of 80.2% and 74.3% were obtained using 0.1% colchicine concentration. All of the 21 entries (5 annual diploids and their induced tetraploids as M. lupulina, M. radiata, M. rijidula, M. truncatula, M. turbinata, and 3 populations of perennial diploid of M. sativa ssp. Caerulea (Karaj1, Karaj2 and Tehran) and their induced tetraploids and 5 commercial tetraploid alfalfa cultivars (Bami, Hamadani, Baghdadi, Ghareyongeh and Renger) grown in pots in a glasshouse experiment using completely randomized design with five replications in 2018 in Ahvaz, Iran. Data were collected for leaf length, leaf width, shoot and root length, seedling weight, shoot weight, root weight, branch number, leaf area index (LAI) and water use efficiency (WUE). Result of mean comparisons between three groups (2x, induced 4x and controls 4x), showed that both groups of induced and natural tetraploids had significantly higher mean values for all the traits except leaf length, shoot length and root length than that for diploids. For the latter traits there were no significant differences between 2x and induced 4x. On the overall, the induced 4x had 60%, 62%, 68%, 65%, 22%, 109% and 47% higher mean values than their parental 2x for seedling weight, shoot weight, root weight, LAI, WUE and branch number, respectively. It was concluded that increasing ploidy level provides plant breeders with a promising tool in the breeding improve new varieties suited for future climate scenarios.

The Genome Copy Number of the Thermophilic Cyanobacterium Thermosynechococcus elongatus E542 Is Controlled by Growth Phase and Nutrient Availability.

The recently isolated thermophilic cyanobacterium Thermosynechococcus elongatus PKUAC-SCTE542 (here Thermosynechococcus E542) is a promising strain for fundamental and applied research. Here, we used several improved ploidy estimation approaches, which include quantitative PCR (qPCR), spectrofluorometry, and flow cytometry, to precisely determine the ploidy level in Thermosynechococcus E542 across different growth stages and nutritional and stress conditions. The distribution of genome copies per cell among the populations of Thermosynechococcus E542 was also analyzed. The strain tends to maintain 3 or 4 genome copies per cell in lag phase, early growth phase, or stationary phase under standard conditions. Increased ploidy (5.5 ± 0.3) was observed in exponential phase; hence, the ploidy level is growth phase regulated. Nearly no monoploid cells were detected in all growth phases, and prolonged stationary phase could not yield ploidy levels lower than 3 under standard conditions. During the late growth phase, a significantly higher ploidy level was observed in the presence of bicarbonate (7.6 ± 0.7) and high phosphate (6.9 ± 0.2) at the expense of reduced percentages of di- and triploid cells. Meanwhile, the reduction in phosphates decreased the average ploidy level by increasing the percentages of mono- and diploid cells. In contrast, temperature and antibiotic stresses reduced the percentages of mono-, di-, and triploid cells yet maintained average ploidy. The results indicate a possible causality between growth rate, stress, and genome copy number across the conditions tested, but the exact mechanism is yet to be elucidated. Furthermore, the spectrofluorometric approach presented here is a quick and straightforward ploidy estimation method with reasonable accuracy.IMPORTANCE The present study revealed that the genome copy number (ploidy) status in the thermophilic cyanobacterium Thermosynechococcus E542 is regulated by growth phase and various environmental parameters to give us a window into understanding the role of polyploidy. An increased ploidy level is found to be associated with higher metabolic activity and increased vigor by acting as backup genetic information to compensate for damage to the other chromosomal copies. Several improved ploidy estimation approaches that may upgrade the ploidy estimation procedure for cyanobacteria in the future are presented in this work. Furthermore, the new spectrofluorometric method presented here is a rapid and straightforward method of ploidy estimation with reasonable accuracy compared to other laborious methods.

Intricate Distribution Patterns of Six Cytotypes of Allium oleraceum at a Continental Scale: Niche Expansion and Innovation Followed by Niche Contraction With Increasing Ploidy Level.

The establishment and success of polyploids are thought to often be facilitated by ecological niche differentiation from diploids. Unfortunately, most studies compared diploids and polyploids, ignoring variation in ploidy level in polyploids. To fill this gap, we performed a large-scale study of 11,163 samples from 1,283 populations of the polyploid perennial geophyte Allium oleraceum with reported mixed-ploidy populations, revealed distribution ranges of cytotypes, assessed their niches and explored the pattern of niche change with increasing ploidy level. Altogether, six ploidy levels (3x−8x) were identified. The most common were pentaploids (53.6%) followed by hexaploids (22.7%) and tetraploids (21.6%). Higher cytotype diversity was found at lower latitudes than at higher latitudes (>52° N), where only tetraploids and pentaploids occurred. We detected 17.4% of mixed-ploidy populations, usually as a combination of two, rarely of three, cytotypes. The majority of mixed-ploidy populations were found in zones of sympatry of the participating cytotypes, suggesting they have arisen through migration (secondary contact zone). Using coarse-grained variables (climate, soil), we found evidence of both niche expansion and innovation in tetraploids related to triploids, whereas higher ploidy levels showed almost zero niche expansion, but a trend of increased niche unfilling of tetraploids. Niche unfilling in higher ploidy levels was caused by a contraction of niche envelopes toward lower continentality of the climate and resulted in a gradual decrease of niche breadth and a gradual shift in niche optima. Field-recorded data indicated wide habitat breadth of tetraploids and pentaploids, but also a pattern of increasing synanthropy in higher ploidy levels. Wide niche breadth of tetra- and pentaploids might be related to their multiple origins from different environmental conditions, higher “age”, and retained sexuality, which likely preserve their adaptive potential. In contrast, other cytotypes with narrower niches are mostly asexual, probably originating from a limited range of contrasting environments. Persistence of local ploidy mixtures could be enabled by the perenniality of A. oleraceum and its prevalence of vegetative reproduction, facilitating the establishment and decreasing exclusion of minority cytotype due to its reproductive costs. Vegetative reproduction might also significantly accelerate colonization of new areas, including recolonization of previously glaciated areas.