Ploidy Manipulation Research Articles

A genetic method to infer ploidy and aberrant inheritance in triploid organisms.

Polyploidy occurs naturally across eukaryotic lineages and has been harnessed in the domestication of many crops and vertebrates. In aquaculture, triploidy can be induced as a biocontainment strategy, as it creates a reproductive barrier preventing farm-to-wild introgression, which is currently a major conservation issue for the industry. However, recent work suggests that triploidisation protocols may, on occasion, produce 'failed triploids' displaying diploidy, aneuploidy and aberrant inheritance. The potentially negative consequences for conservation and animal welfare motivate the need for methods to evaluate the success of ploidy-manipulation protocols early in the production process. We developed a semi-automated version of the MAC-PR (microsatellite DNA allele counting - peak ratios) method to resolve the allelic configuration of large numbers of individuals across a panel of microsatellite markers that can be used to infer ploidy, pedigree and inheritance aberrations. We demonstrate an application of the approach using material from a series of Atlantic salmon (Salmo salar) breeding experiments where ploidy was manipulated using a hydrostatic pressure treatment. We validated the approach to infer ploidy against blood smears, finding a > 99% agreement between these methods, and demonstrate its potential utility to infer ploidy as early as the embryonic stage. Furthermore, we present tools to assign diploid and triploid progeny to families and to detect aberrant inheritance, which may be useful for breeding programmes that utilise ploidy manipulation techniques. The approach adds to the ploidy verification toolbox. The increased precision in detecting ploidy and inheritance aberrations will facilitate the ability of triploidisation programmes to prevent farm-to-wild introgression.

Spontaneous and Chemically Induced Genome Doubling and Polyploidization in Vegetable Crops

Plant ploidy manipulation is often required for breeding purposes. However, there is no comprehensive review covering genome doubling in vegetable crops despite the abundance of data for a large number of vegetable species. Similar to other species, genome doubling is required in vegetable crops to obtain doubled haploids (DHs). It is also utilized for the production of polyploids to overcome interspecific hybrid sterility and improve agricultural traits. Spontaneous haploid genome duplication (SHGD) occurs in many Apiaceae, Brassicaceae, Cucurbitaceae, and Solanaceae crops, allowing for the laborious treatment with antimitotic agents to be bypassed. SHGD mechanisms are not fully understood, but existing data suggest that SHGD can occur via nuclear fusion, endoreduplication, or other mechanisms during microspore or ovule early embryogenic development. Other studies show that SHGD can occur at later developmental stages during extended plant growth in vitro or ex vitro, possibly due to the presence of phytohormones in the medium and/or diploid cell competitive advantage. For unresponsive accessions and species with rare SHGD, such as onion (Allium cepa L.) and beet cultivars (Beta vulgaris subsp. vulgaris L.), antimitotic agent treatment has to be applied. Antimitotic agent application efficiency depends on the treatment conditions, especially the agent concentration and exposure time. Also, plant developmental stage is critical for agent accessibility and plant survival. The existing methods can be used to further improve genome doubling methodology for major vegetable crops and other species.

Genetic diversity studies between Ipomoea wild species and cultivated sweet potato (Ipomoea batatas (L.) Lam.) using SSR markers

Sweet potato (Ipomoea batatas (L.) Lam) is one of the major tuber crops in Asian countries, including India, yet the interrelationship and genetic variability among the cultivar and its crop wild relatives remain obscure. The wild relatives of sweet potato represent a vital source of untapped genetic diversity in the breeding programs. In this study, a set of 12 Simple Sequence Repeats (SSR) markers were utilized to understand the extent of genetic diversity and relationship within and between 20 sweet potato accessions, 23 wild Ipomoea species and two Merremia species for utilization in breeding programmes. The SSR primers produced 84 polymorphic alleles within the range of 105–393 bp, collectively showing an average polymorphism of 94.5 %. The average Polymorphic Information Content (PIC) value of 0.67 showed that all the primers were polymorphic in nature. Similarity coefficient based dendrogram and Principal Component Analysis (PCA) scatter plot clearly differentiated all the cultivars and wild species. The cluster dendrogram analysis revealed the close relationship of cultivars with I. triloba and I. trifida at a similarity index of 0.75. Our analysis showed a high degree of genetic variability within and between hexaploid sweet potato and diploid wild species, whereas lower within the cultivars, which shows that there is a high level of genetic diversity exists within the Ipomoea species for its utilization in breeding programs. Thus, the two wild species, I. trifida and I. triloba could be used in hybridization for improvement of sweet potato through ploidy manipulation.

Genome Doubling of Northern Spicebush, Lindera benzoin L.

Lindera benzoin is a dioecious understory shrub native to eastern North America. Northern spicebush is a beautiful shrub with a natural round shrub shape, golden-yellow fall foliage, attractive bright red drupes, and precocious yellow flowers in early spring; however, its market value as an ornamental value has been overlooked. To improve the ornamental values of this under-cultivated nursery crop, breeding for a better compact form, larger leaves, enlarged flower clusters and fruit, and increased stress tolerances could all be beneficial. Polyploidy manipulation is a valuable method to improve such traits for many ornamental plants. This study established the genome doubling method by oryzalin-infused solid agar treatment on young northern spicebush seedlings. The seedlings of two wild populations in North Carolina were collected and used. A total of 288 seedlings were treated with solid agar containing 150 µM oryzalin for 24, 72, and 120 h. The results were sporadic in their survival ratios and tetraploid conversion ratios between different treatments; however, a total of 16 tetraploid L. benzoin plants were produced in this study. The 24-h treatment showed the optimal result, with 7.1% of total treated seedlings or 15.2% of surviving seedlings converted into tetraploids. Tetraploid plants had visible differences in leaf morphology, a statistically significant enlarged stomata size, and reduced stomatal density compared to diploid plants. This research provides ploidy manipulation information for all future breeding processes of L. benzoin and related species.

Influence of ploidy, genetic and environment on production traits of the Pacific oyster Crassostrea gigas

Pacific oysters, Crassostrea gigas, are one of the most widely farmed shellfish. To meet demand, the advantages of growth and increased sterility are currently being exploited in the commercial culture of triploid C. gigas. However, the extent of the triploid advantage can vary, environmental conditions and rearing methods. In the present study, the performance of diploid and triploid oysters was compared at two sites (Rongcheng and Rushan) to determine whether the triploidization would result in any advantage on production traits that could be utilized for aquaculture of this species. Triploid oysters were also produced using the wild broodstock and selected line to verify whether selectively bred oysters maintain their improved traits after triploidization. In this study, we found that the survival advantage of triploid is “site-dependent” and that Rongcheng's triploids did not exhibit significantly higher cumulative survival than diploids. Notably, incremental survival of triploid oysters was consistently higher than that of diploids during the second summer, which may be related to gonad development and spawning in diploids. At both sites, growth traits (shell height and individual weight) were always higher for triploids than in diploids in both the selected and control groups, which is strong evidence of a superior growth advantage for triploids. In addition, we found that the production performance (growth, survival and yield) of the selected group was consistently better than that of the control group (either diploid or triploid), suggesting that the superior traits of diploids can be converted to triploids through ploidy manipulation. Our results support the notion that yield can be further optimized by reducing mortality through a selective breeding program coupled with triploid production to increase growth rates. This study provides meaningful information on the potential of triploid C. gigas for oyster culture in northern China.

Utilizing method of plant growth regulators induction in processing of polyploidization—A perspective crop development

Nowadays, it seems like human beings are worried about how the world will meet its food security demand urgently when they have faced on rapidly increasing population and combat climate change. Scientists and researchers are indispensably investigating to enhance food sources such developing novel cops with high yield and good quality and even coping with the adverse environment in terms of biotic and abiotic stresses. Thus, there is one valuable method it is believed that should be developed further. In fact, it is believed that human beings should continue using it as soon as possible because it can provide the world with a source of food. Polyploidization introduced by plant growth regulator induction is a good method because it is safety and easy to develop new crops with potential agronomic traits, these polyploidy plants are rare aneuploid and it contains intriguing characteristics of polyploidy plants in adapting to ecological variability. This review sheds light on 1) summarizing molecular mechanism of plant growth regulator induction for plant ploidy manipulation; 2) achieving of polyploidization through plant growth regulator induction; 3) enumerating the perspectives of polyploidization in crop development to cope with climate change. Although the role of phytohormones is underestimated, the effectiveness on physiological level of plants to make polyploidy plants is worth considering and the effects and bio-safety of that on human are also concerned.

A Comprehensive Review on Role of Plant Tissue Culture in Ornamental Crops: Cultivation Factors, Applications and Future Aspects

Ornamental plants are grown largely for their artistic value, floriculturists must prioritize the proliferation and improvement of quality traits, as well as the production of unique diversity. Micropropagation, clonal reliability and conservation are all crucial factors to consider. Application of in vitro techniques in ornamental plant such as in vitro embryo rescue, somatic hybridization, in vitro pollination and in vitro ploidy manipulation but to enhance, techniques like as embryo rescue and somatic hybridization are commonly employed. The creation of synthetic seed allows for season-independent seed producing and long-term seed preservation. Many factors influence ornamental plant tissue culture, including plant genotype, explants type, and the physical environment (light, temperature, humidity, and CO2), in addition to medium composition and growth regulators. We compiled and reviewed an overall update on cultivation factors, application procedures in ornamental plant tissue culture, in vitro plant enhancement approaches and future prospects in this study.

Polyploidy and its relevance in crop improvement

Many areas of research in crop production have been geared towards crop improvement and increased yield. Crop improvement include but not restricted to; plant introduction and acclimatization, domestication, ploidy manipulation (polyploidization), recombinant DNA technology, crossing for superior selection (cultivar development), molecular genetics, etc. Polyploidy is a condition where the genome of an organism has more than the usual number of complete sets of chromosomes and the product of this phenomenon is called a Polyploid. Polyploidy occurs naturally, and can be induced chemically using antimitotic agents or physically using protoplast fusion and temperature shock. It is mostly artificially induced through a process called polyploidization. Polyploids are more advantageous in important plant attributes than the regular diploid. Relative success has been reported in the application of polyploidization for crop improvement which resulted chiefly in increased amount of beneficial secondary metabolites (phytochemicals), larger stomata and leaves,improved adaptation to stress and unfavourable conditions, to mention but a few. Therefore, it is imperative to state that polyploidy is an area of research that has been and will continually be deployed in crop improvement .

Ploidy manipulation for enhancement in fruit quality

Ploidy manipulation for enhancement in fruit quality

Aspects of In Vitro Plant Tissue Culture and Breeding of Asparagus: A Review

In vitro plant tissue culture and biotechnology used to assist and support the development of plant breeding when classical methods of propagation must be accelerated or it was necessary to overcome barriers inaccessible by classical approaches. In asparagus, to improve multiple breeding tasks, a high number of in vitro methods have been used, such as plant regeneration methods through organogenesis, embryogenesis, manipulation of ploidy, protoplast isolation, genetic manipulation (protoplast fusion, genetic transformation), embryo rescue and germplasm preservation (in vitro, in vitro slow growth, cryopreservation). Plant tissue culture methods can overcome multiple problems in asparagus breeding such as, barriers of self and cross-incompatibility between asparagus species through embryo rescue of interspecific hybrids and protoplast fusion or genetic transformation, introgression of new genes, clonal propagation of elite genotypes of asparagus, mass screening, and the generation of haploid and polyploid genotypes, among others, becoming the tool of choice for asparagus breeding programs. Some of these in vitro methods are still under development.

Comprehensive analysis of miRNA-mRNA/lncRNA during gonadal development of triploid female rainbow trout (Oncorhynchus mykiss)

Chromosomal ploidy manipulation is one of the means to create excellent germplasm. Triploid fish could provide an ideal sterile model for searching of a underlying mechanism of abnormality in meiosis. The complete understanding of the coding and noncoding RNAs regulating sterility caused by meiosis abnormality is still not well understood. By high-throughput sequencing, we compared the expression profiles of gonadal mRNA, long non-coding RNA (lncRNA), and microRNA (miRNA) at three different developmental stages between the diploid (XX) and triploid (XXX) female rainbow trout. These stages were gonads before differentiation (65 days post fertilisation, dpf), at the beginning of morphological differences (180 dpf) and showing clear difference between diploids and triploids (600 dpf), respectively. A majority of differentially expressed (DE) RNAs were identified, and 22 DE mRNAs related to oocyte meiosis and homologous recombination were characterized. The predicted miRNA-mRNA/lncRNA networks of 3 developmental stages were constructed based on the target pairs of DE lncRNA-miRNA and DE mRNA-miRNA. According to the networks, meiosis-related gene of ccne1 was targeted by dre-miR-15a-5p_R + 1, and 6 targeted DE lncRNAs were identified. Also, qRT-PCR was performed to validate the credibility of the network. Overall, this study explored the potential interplay between coding and noncoding RNAs during the gonadal development of polyploid fish. The mRNA, lncRNA and miRNA screened in this study may be helpful to identify the functional elements regulating fertility of rainbow trout, which may provide reference for character improvement in aquaculture.

Characteristics of the Diploid, Triploid, and Tetraploid Versions of a Cannabigerol-Dominant F1 Hybrid Industrial Hemp Cultivar, Cannabis sativa 'Stem Cell CBG'.

Hemp (Cannabis sativa L.) has recently become an important crop due to the growing market demands for products containing cannabinoids. Unintended cross-pollination of C. sativa crops is one of the most important threats to cannabinoid production and has been shown to reduce cannabinoid yield. Ploidy manipulation has been used in other crops to improve agronomic traits and reduce fertility; however, little is known about the performance of C. sativa polyploids. In this study, colchicine was applied to two proprietary, inbred diploid C. sativa inbred lines, ‘TS1-3’ and ‘P163’, to produce the tetraploids ‘TS1-3 (4x)’ and ‘P163 (4x)’. The diploid, triploid, and tetraploid F1 hybrids from ‘TS1-3’ × ‘P163’, ‘TS1-3 (4x)’ × ‘P163’, and ‘TS1-3 (4x)’ × ‘P163 (4x)’ were produced to test their fertilities, crossing compatibilities, and yields. The results indicated a reduction in fertility in the triploids and the tetraploids, relative to their diploid counterparts. When triploids were used as females, seed yields were less than 2% compared to when diploids were used as females; thus, triploids were determined to be female infertile. The triploids resulting from the crosses made herein displayed increases in biomass and inflorescence weight compared to the diploids created from the same parents in a field setting. Statistical increases in cannabinoid concentrations were not observed. Lastly, asymmetric crossing compatibility was observed between the diploids and the tetraploids of the genotypes tested. The results demonstrate the potential benefits of triploid C. sativa cultivars in commercial agriculture.

Potential, Challenges and Strategies Involved in Gene Introgression from Wild Relatives of Vegetable Crops: A Review

Wide hybridization is an important plant breeding method as it helps in broadening the gene pool of a crop when the desired variation is not sufficient or absent within the same gene pool. Wild genetic resources are the potential source of desirable genes for various characters of crop plants. It plays a significant role in transferring traits of interest like disease and insect resistance, improved quality, earliness, dwarfness, increased yield, abiotic stress tolerance and manipulation in mode of reproduction etc. in crop plants. It has been instrumental in transferring disease resistance from wild species into many vegetable crops. For example, resistance to late blight, leaf roll and potato virus X, yellow vein mosaic virus and powdery mildew in okra, bacterial wilt, tomato leaf curl virus, early and late blight in tomato has been transferred from wild species into cultivated species. Wild species has been used to improve the quality traits of some crops like carotenoid content in carrot and tomato, starch content in potato etc. Sometimes distant hybridization leads to creation of entirely new crop species. Chromosome elimination technique followed by wide hybridization has been successfully used in the production of double haploids in some crops. However there are some lacunae in wide hybridization such as cross incompatibility, hybrid inviability, hybrid sterility and hybrid breakdown. Some special techniques viz. ploidy manipulation, pistil manipulation, growth regulators treatments, bridge crossing, grafting and embryo rescue have to be adopted to make distant hybridization successful in such cases.

Developing Triploid Maples

Maples are common street and shade trees throughout the temperate zone. They are widely used for their wide range of ornamental traits and adaptability, particularly to urban settings. Unfortunately, some species such as Acer tataricum ssp. ginnala (Amur maple) and A. platanoides (Norway maple) have escaped cultivation to become pests or in some cases threaten native flora. However, these species remain economically important and are still asked for by name. To ameliorate potential future ecological damage from additional escapes, we have been breeding for sterile forms using ploidy manipulation and backcrossing to develop triploids. We began with a series of experiments to develop tetraploids of Amur, Norway, and trident (A. buergerianum) maples. Treatment of seedlings at the cotyledon or first true leaf stage was successful in inducing tetraploids of each species. Mortality, cytochimeras, and tetraploids varied among species. After identifying tetraploids, they were field planted alongside diploid cultivars and seedlings, which served as pollinizers in open-pollination. Seedlings derived from open-pollinated tetraploids were generally found to be a high percentage triploids. Thus far, no Norway or trident maple triploids have flowered but after three years we observed five, 22, and 22 Amur maple triploids flowering over three respective years with no seedlings recovered to date. Further evaluation is required but our findings are encouraging that the triploids we have developed thus far will be sterile and provide new cultivars for nursery growers and land managers.

Fertile Tetraploids: New Resources for Future Rice Breeding?

Ploidy manipulation is an efficient technique for the development of novel phenotypes in plant breeding. However, in rice (Oryza sativa L.), severe seed sterility has been considered a barrier preventing cultivation of autotetraploids since the 1930s. Recently, a series of studies identified two fertile autotetraploids, identified herein as the PMeS (Polyploid Meiosis Stability) and Neo-Tetraploid lines. Here, we summarize their characteristics, focusing on the recovery of seed fertility, and discuss potential future directions of study in this area, providing a comprehensive understanding of current progress in the study of fertile tetraploid rice, a classical, but promising, concept for rice breeding.

A high maternal genome excess causes severe seed abortion leading to ovary abscission in Nicotiana interploidy-interspecific crosses.

Seed abortion and ovary abscission, two types of postzygotic reproductive barriers, are often observed in interspecific and/or interploidy crosses in plants. However, the mechanisms underlying these reproductive barriers remain unclear. Here, we show that the distinct types of seed developmental abnormalities (type I and type II seed abortion) occur in a phased manner as maternal to paternal genome dosage increases and that type II seed abortion is followed by ovary abscission. We revealed that these two types of seed developmental abnormalities are observed during seed development in the interploidy‐interspecific crosses of Nicotiana suaveolens and N. tabacum. Moreover, in the cross showing type II seed abortion, several events, such as changes in abscission‐related gene expression and lignin deposition, occurred in the ovary abscission zone, eventually leading to ovary abscission. Notably, successive increases in maternal ploidy using ploidy manipulated lines resulted in successive type I and type II seed abortions, and the latter was accompanied by ovary abscission. Conversely, both types of seed abortion and ovary abscission could be overcome with a ploidy manipulation technique that balances parental ploidy levels. We thus concluded that a high maternal genome excess cross may cause severe seed developmental defects and ovary abscission. Based on our findings, we propose a model explaining the abortion phenomena, where an interaction between the promotive and inhibitive effects of the parental genomes determines the developmental destiny of seeds.Significance statementWe demonstrate that a stepwise increase in maternal ploidy results in a stepwise increase in seed abortion severity, leading to ovary abscission in plants. We propose a model explaining the abortion phenomena, where an interaction between the promotive and inhibitive effects of the parental genomes determines the developmental destiny of seeds.

In vitro Ploidy Manipulation for Crop Improvement.

In vitro regeneration systems provide a powerful tool for manipulating ploidy to facilitate breeding and development of new crops. Polyploid induction can expand breeding opportunities, assist with the development of seedless triploid cultivars, enhance ornamental characteristics and environmental tolerances, increase biomass and restore fertility in wide hybrids. In vitro ploidy manipulation is commonly induced using antimitotic agents such as colchicine, oryzalin and trifluralin, while many other antimitotic agents have been relatively unexplored. Successful induction requires a synergistic pairing of efficient penetration of the antimitotic agent and may be dependent the length of exposure and concentrations of antimitotic agents, tissue types, and interactions with basal media and plant growth regulators. In vitro conditions vary among taxa and individual genera, species, and cultivars, often requiring unique treatments to maximize polyploid induction. In some taxa, the induction of polyploidy influences in vitro growth, development, and root formation. Here we provide an overview of mitotic inhibitors and their application for in vitro ploidy manipulation for plant breeding and crop improvement.

Use of natural trophic resources by Eastern oysters and Pacific oysters of different ploidy

Ploidy manipulation, such as triploidy, in farmed oysters has been used as a tool to enhance oyster quality throughout all seasons because triploid oysters allocate less energy to gametogenesis, and therefore are deemed better performers than fertile diploids. Nevertheless, scientific reports describing no differences between ploidies and, in certain conditions, disadvantages of triploids are not uncommon. As the use of triploid oyster seed increases culture cost for growers, consistency in performance of triploids is considered to be an important goal. Thus, research to assess how ploidy affects physiological processes underlying oyster performance is fundamental for the aquaculture industry. This work was undertaken to assess if ploidy-based differences in performance in the two most commonly cultivated commercial species of oyster in the United States, Crassostrea virginica and Crassostrea gigas, are associated with filtration, feeding, and metabolism. To test this hypothesis, biodeposition measurements were made with oysters exposed to ambient water conditions at locations and seasons providing a variety of environmental conditions. Oysters did not show differences in filtration and feeding associated with ploidy, but physiological feeding variables fluctuated with environmental characteristics associated with spatial and seasonal differences. A preliminarily test of the hypothesis that differences in energy metabolism may account for differences in performance among ploidy levels indicates that basal metabolic rates of diploid, triploid, and tetraploid Eastern oysters are not different.

High-frequency direct shoot organogenesis from garlic (Allium sativum L.) inflorescence and clonal fidelity assessment in regenerants

Garlic (Allium sativum L.) is an important bulb vegetable with high culinary and medicinal values. It is cultivated by exclusively vegetative propagation which leads to complex virus infection, biodiversity reduction, inhibition of multiple breeding strategies. An efficient and widely applicable shoot regeneration system was established from garlic inflorescence as explant. By optimizing the basal medium type, pH value and explant size, a mean shoot regeneration rate of 97% and mean shoot number of 23.4 per explant were achieved in 14 commercial cultivars with various characteristics and origins. Histological observation revealed that shoots were regenerated through direct organogenesis when meristemoid initiated from subepidermal cells without callus formation followed by periclinal and anticlinal division of epidermal and subepidermal cells. No polymorphic bands were detected by simple sequence repeat (SSR) analysis between regenerants and donor plants. Furthermore, flow cytometry analysis indicated that there was no significant variability of genome size, and all plants maintained their ploidy. These results confirmed the clonal fidelity of regenerants. In conclusion, the present study provides a shoot regeneration system with great potential in micropropagation, germplasm preservation, genetic transformation and ploidy manipulation of garlic. Our study developed a widely applicable procedure for direct shoot regeneration via inflorescence of garlic and ascertained the clonal fidelity of regenerants by using simple sequence repeat (SSR) and flow cytometry.

Genomic-Led Potato Breeding for Increasing Genetic Gains: Achievements and Outlook

Potato is the third most important crop, after rice and wheat, in human diets worldwide. Genetic gains due to its crossbreeding for productivity per se appear to be stagnant in this tetraploid crop. Its genetic enhancement needs to overcome inherent barriers such as ploidy, outcrossing and heterozygosity. Pathogens and pests affect potato because they may infect the entire plant, including stems, leaves and tubers, thus leading to significant tuber yield loss. Hence, host plant resistance breeding remains key for improving the productivity of this crop. This article reviews recent research advances relevant to potato breeding emphasizing genomic resources, methods and tools for genetic analysis, mapping of genes and quantitative trait loci, and genomic prediction of breeding values (or genomic selection) for population improvement. In this regard, association genetics has provided insights onto genetic architecture and inheritance of priority breeding traits, as well as tagging them to DNA markers for their further use as aids for indirect selection. Early research results show the feasibility of genomic selection as a new breeding approach for a tetrasomic polyploid such as potato. This manuscript also highlights the proposed inbred line strategy for producing diploid F1 true potato seed hybrids, and how its use may speed up and increase genetic gains in potato breeding; as well as promoting alleles through gene editing. The paper ends proposing a new interploidy breeding approach considering ploidy manipulations and incorporating genomic selection and gene editing for both population improvement and cultivar development.