Ploidy Variation in Plants

James A Birchler, University of Missouri, Columbia, Missouri, USA

Ploidy variation involves changes in the number of whole sets of chromosomes. Aneuploid variation involves changes in the number of individual chromosomes. The two types of chromosomal changes have different consequences for gene expression patterns.

Secondary article
Article Contents
. Polyploidy . Aneuploidy . Polyteny and Endoreduplication

Polyploidy
Polyploidy is the situation in which there are multiple copies of the genome present in a cell, an individual or a species. The term basic chromosome number (x) refers to the number of chromosomes in a complete set, also known as a genome. The gametic chromosome number (n) is the number of chromosomes in the gametophyte and the gametes. The term haploid refers to the gametophyte and cases in which the gametic number is present in sporophytic tissue. The term monoploid refers to individuals with a single genome in the sporophyte. The most common ploidies are monoploid (1x), diploid (2x), triploid (3x), tetraploid (4x), pentaploid (5x), hexaploid (6x), heptaploid (7x) and octoploid (8x). In diploid species, the haploid has a single genome present and is referred to as a monoploid. Haploids have been found in most taxonomic groups. They are smaller and have reduced oral organs compared to diploids. Haploids can arise via the parthenogenic development of a cell in the megagametophyte, usually thought to be the egg or a synergid (Kimber and Riley, 1963). X-irradiation, delayed pollination and interspecic crosses can also stimulate the production of haploids. In maize, haploids can be generated by a certain inbred line (stock 6) that produces gynogenetic or maternal haploids in several per cent of the progeny (Coe, 1959). The indeterminate gametophyte (ig) mutation conditions paternal or androgenetic haploids (Kermicle, 1969). Monoploids are predicted to give complete genome n complements in the gametes at a frequency of (1 2) , where n is the gametic number of chromosomes. Consequently, most spores (the end product of meiosis) are missing one or more chromosomes and abort. However, normal gametophytes occur at greater frequencies due to spontaneous chromosome doubling in premeiotic tissue, followed by normal meiosis, which produces functional spores. One-third to one-half of all plant species are polyploids of some type; however, in some cases, ancient polyploids have evolved to the point that they behave as a diploid, as is the case with maize. There are two types of polyploids. Autopolyploids contain multiples of the same genome, whereas allopolyploids (also called amphiploids or amphidiploids) are derived from two or more dierent genomes. Allopolyploidy is more common than autopolyploidy.

Triploids can be produced from crosses between tetraploid and diploid plants or by fertilization of diploid female gametes, which result from failed meiosis. The meiotic behaviour of triploids has been studied extensively in Jimson weed (Datura) and in maize (McClintock, 1929; Satina et al., 1938; Punyasingh, 1947). The distribution of chromosomes in meiosis is nearly random. The resulting chromosome numbers range from the 1x to the 2x level. The resulting aneuploidy causes seed abortion, which explains the nearly complete sterility of triploids. Bananas and seedless watermelon derive their sterility from the fact that they are triploids. Most polyploid plants are tetraploids. In allotetraploids, two dierent genomes are present. They arise from crosses between diverged species followed by chromosome doubling. Because the two sets of chromosomes have diverged, the pairing in meiosis occurs between chromosomes that are most similar. This situation results in bivalent formation in prophase of meiosis and consequently in so-called disomic inheritance. In other words, segregation in allotetraploids is the same as in diploid species, although most characteristics are controlled by duplicate genes, because two genomes are present. A recessive allele at both loci must be present for the corresponding phenotype to result. The identical pairs of homologues will preferentially pair with each other in meiosis rather than with the homologous chromosomes, i.e. the partially homologous chromosomes derived from the two progenitor species. In cases in which the chromosome number of a diploid has been doubled, an autotetraploid is formed. When the four chromosomes synapse in meiosis they form a quadrivalent and exhibit tetrasomic segregation. In referring to tetraploid segregation, the homozygous dominant, AAAA, is a quadriplex and the homozygous recessive, aaaa, is a nulliplex. The three types of heterozygotes are: triplex, AAAa; duplex, AAaa; and simplex, Aaaa. Autotetraploid segregation is aected by recombination between the locus being followed and the centromere. When exchange occurs between chromosomes carrying dierent alleles, the resulting products will be two heteroallelic chromosomes and two homoallelic chromosomes. If the heteroallelic chromosomes proceed to the same pole in meiosis I, then in meiosis II both recessive markers could come together in the same meiotic product.
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Ploidy Variation in Plants

Thus, in a triplex heterozygote, AAAa, aa spores can be produced. This process is referred to as double reduction. The genetic distance of a locus from the centromere will determine the level of its double reduction. For genes present near the centromere, a duplex heterozygote (AAaa) produces aa gametes in a 1/6 proportion. Tetraploid plants have altered characteristics compared with the diploid progenitor (Blakeslee, 1941). In most species the leaves, stems, owers and seeds are somewhat larger. A useful means of classication of diploids and tetraploids is the larger size of the guard cells and of the pollen in the latter. While the size of most organs is either larger or of dierent shape in tetraploids compared to diploids, the overall size of tetraploids does not change that dramatically. Cell size correlates strongly with the number of genomes present; thus organs in the higher ploidy are composed of fewer but larger cells. The number of plastids per cell increases with the ploidy level, but individually they remain the same size throughout the ploidy series. Colchicine treatment is often used for the induction of higher ploidy (Blakeslee and Avery, 1937). Heat treatment of embryos immediately following fertilization has been successfully used in maize (Randolph, 1932), wheat, barley, rye and rice. Spontaneous tetraploid sectors occur on diploid plants and can be recovered if owers are included and are self-pollinated. As the ploidy level goes above the tetraploid level in species that are normally diploid, there is a decline in vigour. In Datura, sectors can occur on plants that are 6x and 8x, but whole plants at these ploidies cannot be recovered. In Arabidopsis, hexaploids are fertile and will set seed in crosses with tetraploids, but not when crossed with diploids (Redei, 1964). Interestingly, in nature many species are hexaploid, the most thoroughly studied species being common bread wheat, which is an allohexaploid derived from three related genomes. The relationship of the genomes present to those in progenitor tetraploid and diploid wheats was determined by a series of interspecic crosses followed by an analysis of chromosome pairing in the hybrids (Kihara, 1930; Lilienfeld, 1951).

which are mirror image duplications of a chromosome arm present as an extra chromosome, are even more aected (Blakeslee, 1924; Khush and Rick, 1969), because the equivalent of four doses of the chromosome arm is represented. Monosomics in true diploid species, such as tomato, are very severely aected or are lethal (Khush and Rick, 1966). In maize, where considerable duplication of the genome occurs, monosomics are more readily tolerated (Weber, 1991). In allotetraploid tobacco, monosomics have even less severe eects (Clausen and Cameron, 1944). Still further in allohexaploid wheat, monosomics are quite healthy (Sears, 1944) and indeed nullisomics, i.e. plants missing a pair of homologous chromosomes entirely, can be recovered (Sears, 1953). With each increase in ploidy, the eective change caused by the removal of a single chromosome becomes less.

Dosage analyses of varied ploidy

It has long been recognized that additions or subtractions of chromosomes to the genotype had rather detrimental eects on the vigour or viability of the aicted individual. The explanation, suggested by A. F. Blakeslee based on work with Datura (Blakeslee et al., 1920; Blakeslee, 1921), was that gene products encoded on the varied chromosome were not in the proper balance relative to the gene products from the unvaried portions of the genome. This explanation implicitly assumes that the only changes in gene expression are dosage eects of the varied genes in direct proportion to their copy number. Recent studies, however, indicate a prevalence of trans-acting regulatory dosage eects and suggest a revised explanation for the molecular basis of aneuploid syndromes. The recent data indicate that there are reductions in gene expression in both monosomics and trisomics, leading to the hypothesis that these reductions become rate-limiting on cell and organismal growth. In maize a dosage series of one, two and three copies of the long arm of chromosome 1, which is estimated to span approximately 7% of the genome, produced a similar level of alcohol dehydrogenase 1 and globulin 1, which are both located within this chromosomal segment (Birchler, 1979; Birchler and Newton, 1981). This phenomenon has been referred to as dosage compensation. In addition to dosage compensation, these larger aneuploids produce changes of gene expression in the unvaried portion of the genome (Birchler, 1979; Birchler and Newton, 1981). These trans-acting dosage eects are predominantly of two types. The major one is an inverse eect, resulting in an inverse correlation of the monitored gene expression with the dosage of the aneuploid region. In other words, monosomics with one dose instead of the normal two have an upper limit of 200% of the diploid level of expression. In the three-dose trisomics, there are reductions to 67% of normal. The other type of response

Aneuploidy
Aneuploidy refers to the situation in which cells or individuals deviate by the addition or subtraction of chromosomes from the number in the full genomic complement. In normally diploid species, monosomics have only a single member of a homologous pair, while trisomics have three copies. Typically both monosomics and trisomics are less vigorous than the diploid (Blakeslee, 1934; Khush and Rick, 1966). The monosomic is usually more severely aected than the corresponding trisomic, which is also often reduced in stature. Secondary trisomics,
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Ploidy Variation in Plants

is a direct eect in which there is a direct correlation of gene expression with the dosage of the varied segment, that is, a reduction to 50% of normal in monosomics and an increase to 150% in trisomics compared with the normal diploid. Any one structural gene is typically aected by several aneuploid series (Birchler and Newton, 1981; Guo and Birchler, 1994). The inverse eect appears to be the underlying basis of the dosage compensation found in larger aneuploids (Birchler, 1981). The inverse eect is of the proper magnitude to cancel a gene dosage eect when such a trans-acting regulator and its target structural gene are varied together. The combination of a reduction to 50% resulting from a structural gene dosage eect and a 200% increase of a trans-acting inverse eect in a monosomic results in an expression similar to the diploid. Likewise an increase to 150% in a trisomic is cancelled by a reduction of each of the three copies to 67% of the normal level of expression. Dissection of large aneuploids that exhibit dosage compensation of alcohol dehydrogenase in maize shows that smaller aneuploids surrounding the structural gene exhibit a dosage eect, and another segment of the larger aneuploid produces an inverse dosage eect, on Adh (Birchler, 1981). Thus, the basis of the compensation found in larger aneuploids is the result of a combination of a dosage change of the structural gene and a trans-acting regulatory gene, which inversely aects the structural gene. The changes (or lack thereof) in gene expression in aneuploids described above illustrate a much more complex pattern than mere dosage eects of the varied genes. Indeed the majority of the varied genes are dosage compensated, although a substantial fraction escape this process to produce a dosage eect (Guo and Birchler, 1994). Much more dramatic eects occur in the unvaried portion of the genome. The direct and inverse eects produce decreases and increases in both monosomics and trisomics. In contrast to the extensive changes in gene expression that occur in aneuploids, variation of the whole genome produces less extreme eects (Guo et al., 1996). A study of messenger ribonucleic acid (mRNA) levels from 18 genes in a monoploid, diploid, triploid and tetraploid series in maize showed that the predominant response in a ploidy series is an expression per cell that is directly proportional to the ploidy level (Guo et al., 1996). Indeed, cell size correlates with ploidy, suggesting that the increase in cellular components will produce a larger cell; however, other patterns occur. The major exceptional eect is one in which there is a positive correlation with ploidy that is greater than predicted from the change in ploidy alone. The minor exceptional eect is a negative correlation with ploidy.

Polyteny and Endoreduplication

In specic tissues in some species, deoxyribonucleic acid (DNA) replication has continued without cell division to create nuclei with many copies of the chromosome set. When the entire genome is involved and the replicated chromosomes remain aligned, this situation is referred to as polyteny. Several instances have been documented in plants, most notably in suspensor cells of several species of beans (e.g. Nagl, 1969). In the endosperm of maize, portions of the genome are overreplicated to produce many copies in the cell, a situation referred to as endoreduplication (Gra and Larkins, 1995). In several species that possess small genome sizes, all portions of the chromosomes are endoreduplicated in a developmentally controlled manner (DeRocher et al., 1990).

References
Birchler JA (1979) A study of enzyme activities in a dosage series of the long arm of chromosome one in maize. Genetics 92: 12111229. Birchler JA (1981) The genetic basis of dosage compensation of Alcohol dehydrogenase1 in maize. Genetics 97: 625637. Birchler JA and Newton KJ (1981) Modulation of protein levels in chromosomal dosage series of maize: the biochemical basis of aneuploid syndromes. Genetics 99: 247266. Blakeslee AF (1921) Types of mutations and their possible signicance in evolution. American Naturalist 55: 254267. Blakeslee AF (1924) Distinction between primary and secondary chromosomal mutants in Datura. Proceedings of the National Academy of Sciences of the USA 10: 109116. Blakeslee AF (1934) New jimson weeds from old chromosomes. Journal of Heredity 25: 80108. Blakeslee AF (1941) Eect of induced polyploidy in plants. American Naturalist 75: 117135. Blakeslee AF and Avery AG (1937) Methods of inducing doubling of chromosomes in plants. Journal of Heredity 28: 393411. Blakeslee AF, Belling J and Farnham ME (1920) Chromosomal duplication and Mendelian phenomena in Datura mutants. Science 52: 388390. Clausen RE and Cameron DR (1944) Inheritance in Nicotiana tabacum. XVIII. Monosomic analysis. Genetics 29: 447477. Coe EH (1959) A line of maize with high haploid frequency. American Naturalist 93: 381382. DeRocher J, Harkins KR, Galbraith DW and Bohnert H (1990) Developmentally regulated systemic endopolyploidy in succulents with small genomes. Science 250: 99101. Gra G and Larkins BA (1995) Endoreduplication in maize: involvement of M phase-promoting factor inhibition and induction of S phase-related kinases. Science 269: 12621264. Guo M and Birchler JA (1994) Trans-acting dosage eects on the expression of model gene systems in maize aneuploids. Science 266: 19992002. Guo M, Davis D and Birchler JA (1996) Dosage eects on gene expression in a maize ploidy series. Genetics 142: 13491355. Kermicle JL (1969) Androgenesis conditioned by a mutation in maize. Science 166: 14221424. Khush GS and Rick CM (1966) The origin, identication and cytogenetic behavior of tomato monosomics. Chromosoma 18: 407 420.

ENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net

Ploidy Variation in Plants

Khush GS and Rick CM (1969) Tomato secondary trisomics: origin, identication, morphology, and use in cytogenetic analysis of the genome. Heredity 24: 129146. Kihara H (1930) Genomanalyse bei Triticum und Aegilops. Cytologia 1: 263270. Kimber G and Riley R (1963) Haploid angiosperms. Botanical Review 29: 480531. Lilienfeld FA (1951) H. Kihara: Genome analysis in Triticum and Aegilops. Concluding review. Cytologia 16: 101123. McClintock B (1929) A cytological and genetical study of triploid maize. Genetics 14: 180227. Nagl W (1969) Banded polytene chromosomes in the legume Phaseolus vulgaris. Nature 221: 7071. Punyasingh K (1947) Chromosome numbers in crosses of diploid, triploid and tetraploid maize. Genetics 32: 541554. Randolph L (1932) Some eects of high temperature on polyploidy and other variations in maize. Proceedings of the National Academy of Sciences of the USA 18: 222229. Redei G (1964) Crossing experiences with polyploids. Arabidopsis Information Service 1: 13. Satina S, Blakeslee AF and Avery AG (1938) Chromosome behavior in triploid Datura. III. The seed. American Journal of Botany 25: 595602.

Sears ER (1944) Cytogenetic studies with polyploid species of wheat. II. Additional chromosomal aberrations in Triticum vulgare. Genetics 29: 232246. Sears ER (1953) Nullisomic analysis in common wheat. American Naturalist 87: 245252. Weber DF (1991) Monosomic analysis in maize and other diploid crop plants. In: Gupta PK and Tsuchiya T (eds.) Chromosome Engineering in Plants: Genetics, Breeding and Evolution, Part A, pp. 181209. Amsterdam: Elsevier.

Further Reading
Haldane JBS (1930) The theoretical genetics of autopolyploids. Journal of Genetics 22: 359372. Mather K (1936) Segregation and linkage in autotetraploids. Journal of Genetics 32: 287314. Muntzing A (1933) Hybrid incompatibility and the origin of polyploidy. Hereditas 18: 3335. Soltis DE and Soltis PS (1993) Molecular data and the dynamic nature of polyploidy. Critical Reviews in Plant Sciences 12: 243273. Stebbins GL Jr (1947) Types of polyploids: their classication and signicance. Advances in Genetics 1: 403429.

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