J. Phycol. 46, 860–867 (2010) Ó 2010 Phycological Society of America DOI: 10.1111/j.1529-8817.2010.00874.x REVIEW WHAT DO WE KNOW ABOUT CHAROPHYTE (STREPTOPHYTA) LIFE CYCLES?1 David Haig2 Department of Organismic and Evolutionary Biology, Harvard University, 26 Oxford Street, Cambridge, Massachusetts 02138, USA The charophyte algae are the closest living rela- Toward the end of the century, zoologists discov- tives of land plants. Their life cycles are usually ered that ova and sperm had half the number of characterized as haploid with zygotic meiosis. This chromosomes of somatic cells, and botanists deter- conclusion, however, is based on a small number of mined that the sexual generation of land plants had observations and on theoretical assumptions about half the number of chromosomes of the asexual what kinds of life cycle are possible. Little is known generation (Farley 1982). Therefore, the life cycles about the life cycles of most charophytes, but unu- of both plants and animals were seen to alternate sual phenomena have been reported in compara- between two nuclear phases with a 2-fold difference tively well-studied taxa: Spirogyra and Sirogonium are in chromosome number. The reduction in chromo- reported to produce diploid gametes with synapsis some number was immediately followed by syngamy of homologous chromosomes before fusion of in animals. Therefore, all cells except gametes were gametic nuclei; Closterium ehrenbergii is reported to diploid. However, in land plants, chromosome undergo chromosome reduction both before and reduction and syngamy were both followed by a ser- after syngamy; and zygotes of Coleochaete scutata are ies of cell divisions such that the life cycle contained reported to replicate their DNA to high levels both multicellular haploid and diploid phases. before a series of reduction divisions. All of these The recognition of an alternation of haploid and phenomena require confirmation, as does the con- diploid phases changed the way the alternation of ventional account. generations was conceptualized. The sexual (game- tophyte) generation of land plants was haploid, and Key index words: alternation of generations; Chara; the asexual (sporophyte) generation was diploid, charophyte; Closterium; Coleochaete;meiosis;Spi- but this close correspondence did not apply to the rogyra;syngamy sexual and asexual generations of algae. For exam- Abbreviations: C, the amount of DNA in an unre- ple, zoospore-producing and gamete-producing plicated haploid genome; G, the amount of DNA thalli of Coleochaete both develop from zoospores in a gametic nucleus immediately prior to karyog- and presumably have the same chromosome num- amy ber. Before the discovery of an alternation of nuclear phases, these thalli were viewed as repre- senting an alternation of asexual and sexual genera- Nineteenth-century botanists recognized an alter- tions, but since the discovery, the two kinds of thalli nation of asexual and sexual generations. An asex- have increasingly been conceptualized as a single ual generation produced offspring that developed kind of thallus, a haploid (gametophytic) genera- from a single cell (spore), whereas a sexual genera- tion with alternative reproductive modes (Haig tion produced offspring that developed from the 2008). union of two cells (gametes). For land plants, the When botanists set out to understand how chro- two kinds of generations had contrasting morpho- mosome number changed during algal life cycles, logies, but, among algae, sexual and asexual genera- most implicitly assumed that algae would exhibit a tions were sometimes indistinguishable apart from similar alternation between haploid and diploid their reproductive structures. For example, vegeta- nuclear phases (Renner 1916). Therefore, a life tive thalli of Coleochaete sometimes reproduced by cycle would be defined by whether mitotic divisions zoospores, in which case the thallus was an asexual followed the reduction division, creating a multicel- generation, and sometimes by gametes, in which lular haploid phase, and whether mitotic divisions case the thallus was a sexual generation (Haig followed syngamy, creating a multicellular diploid 2008). phase. Algal life cycles were expected to follow ‘‘rules’’ established in land plants, and their descrip- tions were often shoe-horned into an ill-fitting ter- minology suited to land plants rather than 1Received 25 September 2009. Accepted 26 February 2010. 2Author for correspondence: e-mail [email protected]. described in their own terms. 860 CHAROPHYTE LIFE CYCLES 861 One of the principal reasons for studying algal (a) life cycles was the hope that such studies would shed 4C light on the origin of the sporophyte of land plants. Interpretation, however, was hampered by uncer- 2C tainty about algal phylogeny. Molecular phylogenies now unequivocally establish that embryophytes were 1C derived from within the charophyte algae (Karol et al. 2001, Qiu et al. 2007; I use ‘‘charophyte’’ in the broad sense of Lewis and McCourt 2004 and use (b) ‘‘stonewort’’ to refer to the members of the Cha- 4C rales). Charophyte algae are generally believed to 2C possess life cycles in which vegetative cells are hap- loid and zygotes are the only diploid cells. There- 1C fore, such a life cycle has been inferred to have been present in the algal ancestors of embryophytes and to support the ‘‘antithetic’’ theory for the ori- (c) gin of sporophytes (Blackwell 2003, Donoghue 4C 2005, Haig 2008, Becker and Marin 2009). Common knowledge is sometimes collective mis- 2C information, and it is worthwhile to occasionally subject what everybody knows to critical reappraisal. 1C This paper reviews more than a century of studies Fig. 1. Conventional interpretations of algal life cycles. Each of charophyte life cycles and finds few observations life cycle is represented as proceeding from a zygotic nucleus that have been replicated by multiple observers. I (square) to a gametic nucleus (circle). Rising solid lines repre- conclude that something like the standard interpre- sent the increase in DNA content during a cell cycle. Dotted lines tation may be true for some forms, although sup- represent the decrease in DNA content per nucleus at cell divi- sion. Mitosis is associated with a halving of DNA content at a sin- porting evidence is flimsy, and that the diversity of gle division, whereas meiosis is associated with a reduction to charophyte algae is likely to encompass a corre- one-quarter over the course of two divisions. (a) Life cycle with sponding diversity of life cycles, some of which will zygotic meiosis. DNA content is raised to the 4C level after syn- not fit neatly into conventional categories. gamy. Meiosis reduces DNA content to the 1C level. Haploid nuclei alternate between the 1C and 2C level of DNA. (b) Life Changes in DNA content. The default assumption, cycle with multicellular diploid and haploid phases. After syn- since the discovery of an alternation of nuclear gamy, DNA content of diploid nuclei alternates between the 2C phases, has been that sexual life cycles operate at and 4C levels. Meiosis reduces DNA content to the 1C level. Hap- two ploidy levels. The amount of DNA in a nucleus loid nuclei alternate between the 1C and 2C levels. (c) Life cycle doubles during the course of a conventional cell with gametogenic meiosis. Diploid nuclei alternate between 2C and 4C levels. Meiosis produces 1C gametic nuclei. cycle and then is reduced by half at mitosis. There- fore, the alternation of haploid and diploid phases has been assumed to imply an alternation between 4C and 2C levels in diploids, and between 2C and et al. 1994, Garbary and Clarke 2002). Among 1C levels in haploids, where C is the amount of charophyte algae, 8-fold or higher reduction has DNA in an unreplicated haploid genome. If meiosis been reported in the zygospore of Coleochaete immediately follows syngamy, then the zygote is the (Hopkins and McBride 1976) and in somatic cells only diploid cell in the life cycle and all other cells of Chara (Shen 1967). are haploid (Fig. 1a). If meiosis and syngamy are As I do not wish to assume that charophyte life both followed by mitosis, then the life cycle contains cycles are ‘‘well-behaved,’’ I will use 2G to refer to both vegetative haploid and vegetative diploid cells the amount of DNA in a zygotic nucleus immedi- (Fig. 1b). If meiosis immediately precedes syngamy, ately after karyogamy and 1G to refer to the amount then gametes are the only haploid cells and all of DNA in gametic nuclei immediately prior to kary- other cells are diploid (Fig. 1c). ogamy. The latter is not necessarily equal to 1C, the Haploid–diploid alternation requires mechanisms amount of DNA in an unreplicated haploid gen- for doubling DNA content (DNA replication and ome, because gametic DNA may have replicated syngamy) and mechanisms for halving DNA content prior to karyogamy, or gametes may contain more (meiosis I, meiosis II, and mitosis) that are suffi- than one haploid genome. I will use ‘‘meiosis’’ to cient, in theory, to allow life cycles that encompass refer to a sequence of two (or more) divisions with- more than three levels of DNA content per nucleus. out intervening synthesis of DNA. Indeed, 8-fold or greater reduction in chromatin Charophyte life cycles. Little is known about the life content has been reported in nuclear cycles from cycles of most charophyte algae. Sexual reproduction pyrsonymphids (Hollande and Carruette-Valentin has never been described in significant taxa such 1970), radiolaria (Le´cher 1978), red algae (Goff as Mesostigma viridis and Chlorokybus atmophyticus. and Coleman 1986), and brown algae (Garman Syngamy has been reported in Chaetosphaeridium 862 DAVID HAIG globosum (Thompson 1969) and Klebsormidium flacci- cation), with DNA synthesis occurring in 0.5G dum (Wille 1912), but these descriptions provide no gametic nuclei to bring them up to 1G before kary- information about germination of zygospores or ogamy (Fig. 2a). Gametic nuclei contain bivalents, about chromosome numbers at different life-history each of four chromatids, before karyogamy.