Jasen Wise

Final Draft

Population Genetics

 

Pteropsidia Polyploidy

"Under the microscope you can see the strange and beautiful figures which the chromosomes execute to ensure proper distribution of genes."

- Julian Huxley, 1953

Introduction

Despite the absence of a universally accepted definition of a species, most would agree that definable species exist within biological systems.  Evolution, a key component to the origins of species, is an ongoing and continuously changing force upon a population. Even though this progression is continuous, it is still possible to define distinct units where boundaries can be delineated and differentiated from one another. Around 2 million species have been identified through various comparisons such as morphology, behavior, genetics, and ecology (Haufler and Hooper, 2000). Ferns present prime examples of how speciation through the mechanisms of ecology and genetics have lead to a successful, diverse, and genetically stable group of plants collectively known as division Pteropsida. 

The division Pteropsida consists of the true ferns. There are anywhere between 10,000 to 15,000 species of ferns divided into 40 families.  Two common examples of true ferns include the genus Polystichum, which includes the Christmas Fern and the genus Asplenium, which contains the Ebony Spleenwort (Cobb,1963). Ferns are among the oldest extant land plants and appear in the fossil record for the past 360 million years (Nicholls, 1998). Such long-term success can be partially attributed to unique fern reproduction and genetic systems.  In order to understand the advantages of polyploidy used by ferns to be successful and widely distributed throughout the world it is necessary to  review the mechanisms by which polyploids are formed,  and types of polyploids.  It is also important to evaluate the  advantages pertaining to inbreeding depression,  buffering effects, and colonization in reference to the challenges that polyploids must face when establishing in new areas. 

The Unique Fern Life Cycle

Ferns exist worldwide and all share an unusual form of reproduction through the alternation of generations. The completion of each generation involves the cycling of two very different morphologies and chromosome compliments. Unlike angiosperms, organism that reproduce via seeds, ferns produce many tiny spores, each of which forms a small haploid (n) plant called a gametophyte. Gametophytes produce male and female gametes that form a sporophyte (2n). The form of the fern familiar to most is called the sporophyte.  The sporophyte forms and matures, starting the cycle again (Cobb, 1963). Having a haploid gametophyte and a diploid sporophyte allows for many variations in fern chromosome numbers. Multiple ploidys (variations in chromosome number) sometimes occur within species, which is termed a cytotype (Soltis and Soltis, 2000). The acquisition of another, or more than one, extra genome is referred to as polyploidy. 

Polyploidy

An organism may be defined as a polyploid if it has more than two haploid (>2n) sets of chromosomes. In mammals, polyploidy is devastating, however it occurs often in the plant kingdom. There are two main types of polyploidy, autopolyploidy and allopolyploidy.  Autopolyploidy is polyploidy in which all the chromosomes originate from the same diploid parent species. In the experiments of DeYoung et al., (1997) an autotetraploid fern, Ceratopteris, was identified by the formation of multivalents at metaphase.  Autopolyploids form multivalents in mitosis when the homologous chromosomes pair up at metaphase making them easily distinguishable from their diploid counterparts (DeYoung et al, 1997).

 Allopolyploidy is a polyploidy in which the sets of chromosomes are from different species. Usually hybrid plants (n + n') are not fertile because proper pairing of chromosomes does not occur in meiosis. Sometimes the chromosome number spontaneously doubles leaving tissues with 2n + 2n'.  If this tissue is germ tissue, tissue than can give rise to haploid tissue via meioses, the result can be gametes with the n + n' chromosome complement.  The fusion of  two of these gametes results in  an allopolyploid plant with a viable chromosome complement (2n + 2n'). An example of an allopolyploid fern is the Green Mountain Maidenhair fern, Adiantum viridimontanum. This fern was formally described in 1991.  The evidence for the allotetraploid cross between common Woodland Maidenhair (Adiantum pedatum) and Aleutian Maidenhair (A. aleuticum) is based on genetic evidence (bivalent mitosis) (Ruesink, 2001).

G. Ledyard Stebbins (1950) proposed an additional category of polyploidy called segmental allopolyploidy. Segmental allopolyploids are formed when the parent diploid is slightly different between its chromosome sets (a vs a'), but its parents were similar enough to be identified as the same species.  A chromosome doubling occurs and a new segmental allopolyploid is formed.  Some multivalent pairings occur at meiosis but usually with the consequence of a lower fertility rate than strict allopolyploids. They can also form some fertile hybrids through backcrossing with autopolyploid derivatives of either parental species.

There are several mechanisms by which polyploids arise. Three examples are somatic doubling, gametic nonreduction and triploid bridges.  Somatic doubling occurs at the zygotic, embryonic, or meristematic stages of a plant’s life cycle. Polyploid offspring can be generated from the production of polyploid tissues. Although many examples of polyploidy via somatic doubling have been reported, this mechanism now seems less common than gametic nonreduction, or the production of unreduced gametes, as a means of polyploid formation in natural populations (Harlan and deWet, 1975).   Unreduced gametes have been reported in a number of species, most notably those that also produce polyploids (Ramsey and Schemske, 1998). Both auto- and allopolyploids can arise in one step after unreduced gamete formation by the union of two unreduced gametes from the same plant.   

Alternatively, the production of either an auto- or allotetraploid may involve a ‘triploid bridges”.   Tetraploids are formed by triploid intermediates formed within a diploid population by backcrossing to diploids or by self fertilization of the triploid. This two-step method has been considered a significant pathway to polyploid formation (Harlan and deWet, 1975), but some (Bretagnolle and Thompson, 1995) suggest that the one-step process involving the union of two unreduced gametes may be more common.  Ramsey and Schemske (1998) concluded that the triploid intermediates may be more significant in the formation of tetraploids since triploids could  be more reproductively viable than originally thought. 

The Advantages of Polyploidy

The main advantage of polyploidy is the production of heterozygotes.  Typically the diploid Mendelian cross of AA and A’A’ would produce a 1:2:1  genotypic ratio (1 homozygous AA, 2 heterozygous AA’ and, 1 homozygous A’) of progeny. However, in a polyploid species the frequency of heterozygotes are much greater. A tetraploid, for example, cross of AAAA and A’A’A’A’ would result in a genotypic ratio of 1:32:1 (Stebbins, 1947). Heterozygotes offer many advantages including buffering effects, protection from inbreeding depression, and the unidirectional introgression phenomenon (Soltis and Soltis, 1995).

Buffering Effects

Tetrasomic inheritance has a buffering effect on intermediate genotypes, an effect of great adaptive value to a population (Stebbins, 1950). This production of nearly all heterozygous ferns is an advantage because it makes the appearance of deleterious homozygous alleles rare because of the masking effect of additional genomes (Soltis and Soltis, 2000).

Two closely related tetraploid species, for example, that hybridize have five possible genotypic progeny: (aaaa), (aaaa'), (aaa'a'), (aa'a'a'), and (a'a'a'a').  Therefore, it can be expected that those with more "a" alleles would be well suited for an area where that allele thrived.  Likewise it can be expected that a greater composition of the a' allele would be best suited for areas where a' thrived.  This combination of recombination, natural selection, and genetic segregation is capable of producing an entire spectrum of genotypic populations that would be compatible with any region of intermediate environmental conditions (Stebbins, 1971).

Inbreeding

An isolated plant is forced to undergo self fertilization or selfing. In this situation the frequency of deleterious homozygous alleles increases leading to an overall reduction in the colony’s overall fitness.  This inbreeding depression is defined as a reduction in fitness and vigor of individuals as a result of increased homozygosity through inbreeding in a normally outbreeding population. For example, an isolated colony of diploid plants would experience an increase in the number of deleterious homozygous alleles and have increased progression towards fixation or even loss of certain alleles.  Studies have found that ferns actually have increased production of normal sporophytes when they are isolated and forced into self fertilization (Soltis and Soltis, 2000). 

 

When considering a single allele at a single locus, the effects of inbreeding depression do not appear to a problem of great importance. The problem is escalated when all the loci are taking into consideration (Lynch, 1995).   For example, examinations of lower organisms and other plants estimate about 100 deleterious alleles to be present in individuals when all genetic loci are examined.  Individually these alleles produce only a small reduction in fitness ( 2%) when homozygous.  However, the combination  of all 100 homozygous loci accumulated through inbreeding could potentially reduce the fitness of the organism on the order of 200%!  This is enough to "kill" the individual two times over (Lynch, 1995)

Despite the disadvantages of inbreeding,  polyploid ferns have grown to favor selfing as a form of reproduction since polyploid protections against inbreeding depression. Two studies of inbreeding depression in diploid and tetraploid ferns show this tendency. In Phegopteris 30-60% of all selfed gametophytes from the diploid race produced normal sporophytes. However 100% of the selfed gametophytes of the tetraploid race formed sporophytes.  Another example of selfing preference is present in Lepiosorus.  Here, only 4% of the selfed diploid race produced normal sporophytes when nearly 100% of the selfed tetraploid race produced normal sporophytes (Soltis and Soltis, 1995).

Unidirectional introgression

Unidirectional introgression is an important advantage in that the tendency occurs to assist chromosomal segregation in helping ferns colonize into new areas.  A combination of hybridization and natural selection on the backcrossed and better suited progeny, produces a tendency for the diploids to form tetraploids or other higher number systems.  When triploids occur from this diploid–tetraploid cross, many are incompatible as a triploid hybrids.  Therefore there is a tendency for these variants to form tetraploids (Sebbins, 1971).

Colonization of New Areas via Polyploidy

It is estimated that as much as half of all angiosperms and nearly all (95%) pteridophytes have multiple polyploid races, also called cytotypes (Soltis and Soltis, 2000). One advantage of cytotypes is that they can exist in a region in a broad range. The Appalachian Bristle Fern (Trichomanes boschianum), for example, has two distinct cytotypes or ‘genetic races’ found at the eastern and western ends of its range (Farrar, 1993). Polyploid fern populations are scattered throughout the world. An examination of the Rockfern, Asplenium ceterach, has shown several cytotypes spread throughout Europe. Tetraploid populations were found throughout the continent , but diploid populations were scarce and localized in the Pannonian-Balkan region.

 Hexaploids found in southern Mediterranean populations.   A separate polyploid complex in Greece had diploid, tetraploid and hexaploid populations with two native haplotypes. These findings suggest that populations in the southern Mediterranean have been persistent for a long time.  Though examination of cpDNA (chloroplast DNA), three haplotypes were found to be common among the tetraploids of Spain and Italy (Trewick and Morgan-Richards, 2003). Another example of regional variants are the Adiantum raddianum ferns of the Nilgris, in South India.  A diploid (n=57), tetraploid (n=114), and even an octoploid (n=228) were observed in that region (Bir and Irudayaraj, 2001).

 

Polyploid Potential for Evolution and Success

            Observations that new species arise from the formation of polyploids, like the previously mentioned Green Mountain Maidenhair fern, have not gone unnoticed. However,  environmental factors play a roll in the success of these newly formed polyploids. Some suggest that the evolutionary potential for polyploids depends on a number of factors. Mainly, how the genes on the new chromosomes are distributed (Soltis and Soltis, 2000).

 The success of the polyploid may depend partially on the particular parental DNA sequence, paternal or maternal, and how it interacts with the organellear genomes.  The location of the sequences and their coding or non-coding function are also have an effect.  Sequences near telomeric, centromeric, or heterochromatin will all behave differently than other regions of the chromosome. The question remains to what extent the differentiation between parents affects the genetic make-up of the organism (Soltis and Soltis, 2000).

            The effects of hybrid vigor do not always create a super plant that will dominate the region.  More often , polyploid plants must over come barriers to establish populations such as the minority cytotype exclusion.  Minority cytotype exclusion is at work when newly-formed outcrossing polyploids have few potential mates when a polyploid ends up among many diploids. This is detrimental because the tetraploid may waste all of its gametes on sterile matings. The success that polyploids demonstrate is biased because we observe the examples that have taken advantage of the numerous benefits of polyploidy and have overcome polyploidy barriers to establish themselves as species .

Conclusion

            Ferns have taken full advantage of their polyploid gifts and established themselves throughout the world.  They have been successful in isolated populations by using the advantages of polyploidy to quench inbreeding depression and also evolve new species. They have overcome the hardships of minority cytotype exclusion and moved into new regions by unidirectional introgression.  They have also taken advantage of their hybrid formation for the function of buffering effects.  The unique reproduction abilities and genetic advantage of polyploid cytotypes in the Pteropsidia, have enable the fern to evolutionarily adapt and maintain success for the past 400 million years.

           


References

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“We have the receipt of fern seed – we walk invisible.”

 

William Shakespeare –Henry IV