Karyological analysis of order testudines 2016
Introduction
Here discuss about the biology that lead to studing the number of chromosome in the order tetudines and inheritance of the chromosomes in the offsprings and also studying of molecular structure and function of gene ,gene behavior of a cell or organism is the genetics
And also studying gene distribution,variation in population
Cytogenetic
that is branch of genetics that considered with studing the structure and function of cell specially chromosomes it also give acloser and amore comprehensive studying on changes in structure and number of chromosomes from one organism to another and this fact was used in what is called cytotaxonomy.
Cytotaonomy
Is a branch of science that classifies the livinh organisms based on cytological studies (number of chromosomes meiosis behavior)
Helps to stablish relationships between the different organism one of the methods of karyotyping
Karyotype
Is Method where total set of chromosomes of an organism is viewed under microsocope where the number of chromosome along with their length position of centromeres Banding pattern any differences between sex chromosomes and any others physical CHARACTERISTIC is observed (king, stansfield and mulligan 2006)
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Karyological analysis of order testudines 2016
AIM OF WORk
Karyological data are available for 55% of all cryptodiran turtle species including members of all but one family. Cladistic analysis of these data as well as consideration of other taxonomic studies, lead us to propose a formal classification and phylogeny not greatly different from that suggested by other workers. Werecognize 11 families and three superfamilies. The platysternid and staurotypidturtles are recognized at the familial level. Patterns and models of karyotypic evolution in turtles are reviewed and discussed.
RESONE FOR STUDING TURTLE
The history of where turtles are found is an important record for conservation and preservation efforts and an invaluable resource for anyone interested in turtle research. If you have ever wondered which turtles are found where you live, you are interested in turtle research.
Because of diversity of this species of turtle and according to a lots of scientists that made research on order testudines that will disscuss in the following , so that I want studing this research
Within the conversation community, turtles are considered to be in crisis situation , brought about by human activities {reviewed in van Dijk et al ., 2000; turte conversation fund 2002}
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Karyological analysis of order testudines 2016
Currently , out of 200 species of fresh water turtle and tortoises listed by the world wide union for te listed as he conversation of nature {iucn} in their red list {IUCN, 2006} , 24 are
Listed as critically endangerd
HOWEVER, about 100 species of fresh water turtles are not listed by IUCN in the red list , either because they are mor common or have not yet been evaluated for listing this mean that at least about 42% of freshwater turtles and tortoises are considered to be facing a high risk of extinction , and are in need of urgent conversation action
Turtles have been prized as pets or killed for commercial products and although some of this trade is met by commercial farms illegal harvest from the wild occurs on a broad scale
In many {thorbjarnarson et al.,2000} Morphology and taxonomy Introduction of turtles
A turtle is an animal in armor. Much of its body lies within a protective shell, which has openings for the turtle's four chunky legs, short tail, and head. When danger threatens, many turtles pull legs, tail, and head into the shell. But unlike some animals that live in shells, such as hermit crabs and snails, a turtle cannot crawl out of its shell. The shell is part of the turtle's body.
All turtles belong to the class of backboned animals known as reptiles. This class also includes snakes, lizards, and crocodiles. Turtles are the oldest group. The first turtles crawled about on earth more than 250,000,000 years ago. Turtles have changed very little since that time.
Turtles are found in almost all temperature and tropical regions of the world. Many turtles spend all or most of their lives in fresh water. They may live in swamps, ponds, running streams, or even roadside ditches.
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Karyological analysis of order testudines 2016
They come up on dry land to sun themselves or to lay eggs. Other turtles live completely on land. Still others live in warm seas, sometimes following warm currents far northward.
The name "turtle" is often used to identify those animals that live in water. The name "tortoise" frequently refers to a turtle that lives on land. The American Indian name "terrapin" usually refers to small freshwater turtles, especially those used for food. But these groupings are not strictly scientific. In this article, all of these animals will be referred to generally as turtles, though the proper name for a specific animal, such
as Galápagos tortoise, will be used.Fig1
)Fig.1)
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General characteristics (around300species)-
Rigid shell enclosing the soft organs (Fig 2) -Carapace= dorsal part -Plastron= ventral part -Shell is composed of dermal bony elements covered by keratinous scutesor leathery skinthe shell incorporates ribs, vertebrae, portions of pectoral girdle-Plastron can be rigidor hinged -Shell shape –ranges from domed(in terrestrial species) Flat to hydrodynamic shaped(aquaticand marine species
(fig3)
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Karyological analysis of order testudines 2016
pads (terrestrial Absence of teeth (keratinous beakinstead) -Freshwater species carnivorous, omnivorous, or herbivorous; terrestrial usually herbivorous. -Limb structure –flippers(marine species), webbing between digits (freshwater species), stout limbs with thickened species)
"classification"
Kingdom Animalia animals
Phylum Chordata chordates
Subphylum Vertebrata vertebrates
Superclass Gnathostomata jawed vertebrates
Class sauropsida
Subclass anapsida
Order testudines
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Suborder Cryptodira
Super family testudinidae
Family Emydidae (Pond Turtles/Box and Water Turtles)
Family Testudinidae (Tortoises)
Family Geoemydidae (Bataguridae) (Asian River Turtles, Leaf and RoofedTurtles, Asian Box Turtles)
Family Platysternidae (Big-headed Turtles)
Family chelydridae ( snapping turtle)
Superfamily Trionychoidea
Family Carettochelyidae (Pignose Turtles) Family Trionychidae (Softshell Turtles)
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Superfamily Kinosternoidea
Family Dermatemydidae (River Turtles)
Family Kinosternidae (Mud and Musk Turtles)
Superfamily Chelonioidea
Family Cheloniidae (Sea Turtles)
Family Dermochelyidae (Leatherback Turtles)
Suborder Pleurodira
Family Chelidae (Austro-American Sideneck Turtles) Superfam. Pelomedusoidea
Family Pelomedusidae (Afro-American Sideneck Turtles) Family Podocnemididae (Madagascan Big-headed and American Sideneck River Turtles)
Sauropsida ("lizard faces") is a group of amniotes that includes all existing reptiles and birds as well as their fossil ancestors and relatives. Sauropsida is distinguished from Synapsida, which includes mammals and their fossil ancestors. This clade includes Parareptilia and other extinct clades. All living sauropsids are members of the sub-group Diapsida.
An anapsid is an amniote whose skull does not have openings near the temples.[1] Traditionally, the Anapsida are the most primitive subclass of reptiles, the ancestral stock from
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Karyological analysis of order testudines 2016 which Synapsida and Diapsida evolved, making anapsids paraphyletic. It is however doubtful whether all anapsids lack temporal fenestra as a primitive trait, or whether all the groups traditionally seen as anapsids truly lacked fenestra DeBraga, M. (1996)
Temple indicates the side of the head behind the eyes. The bone beneath the temporal bone as well as part of the sphenoid bone.
(Fig5)
Cryptodira is a suborder of Testudines ;Zug, G. R. 1966) that includes most living tortoises and turtles. Cryptodira differ from Pleurodira (side- neck turtles) in that they lower their necks and pull the heads straight back into the shells, instead of folding their necks sideways along the body under the shells' margins. They include among their species freshwater turtles, snapping turtles, tortoises, soft-shell turtles, and sea turtles.
Two circumscriptions of the Cryptodira are commonly found. One is used here; it includes a number of primitive extinct lineages known only from fossils, as well as the Eucryptodira. These are, in turn, made up from some very basal groups, and the Centrocryptodira contain the prehistoric relatives of the living cryptodires, as well as the latter, which are collectively called Polycryptodira. (Gaffney, E. S. 1975)
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The family Testudinidae contains approximately 11 genera and 40-50 species the plastron is usually without a hinge carapace is domed,
Ornat-box turtle (family emydidae) (Fig6)
Adaptations for terrestrial life include thick, elephantine rear legs, short, web-less feet, and short digits. The forelegs usually have heavy scales on the anterior surface. Tortoises can be diagnosed by the lack of glands in the axillary and inguinal regions and the presence of only four digits on the rear feet.( Gray, J. E. 1870)
The Testudinidae are most closely related to the pond turtles (Emydidae) and are included along with that family in the Testudinoidea. the Emydidae is split into two families- Emydidae and Bataguridae
The reduced volume of a fusiform body means sea turtles can not retract their head, legs, and arms into their shells, like other turtles can. the number of and shape ofscutes on the carapace, and the type of inframarginal scutes on the plastron.There are seven extant species of sea turtles: the green, loggerhead, Kemp's ridley, olive ridley, hawksbill, flatback, and leatherback. (Nakamura ;1949)sea turtles
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Karyological analysis of order testudines 2016 have a more fusiform body plan than their terrestrial or freshwater counterparts
Chelonia mydas green sea turtle (Fig.7)
Trionychia is a superfamily of turtles which encompasses the species that are commonly referred to as softshelled turtles as well as some others. They are found throughout the temperate regions of the world.( Jordan,1975)
It traditionally consisted of a single family, two subfamilies, and 14 genera. However, more recently it was realized that the supposed "Kinosternoidea" are actually early offshoots of the Trionychoidea and not as closely related among (Pablo A. Martinez ; et al 2009)each other as it was believed. These two families lack the characteristic trionychoid apomorphies, but possess some highly derived characters of their own, which they moreover evolved independently from each other. (Frair, W. 1972)
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apalone spinifera turtle (Fig.8)
Kinosternoidea is a superfamily of aquatic turtles, which included two families: Dermatemydidae, and Kinosternidae.
These are nowadays usually considered independent families of the Trionychia, among which they represent ( Laurin, etal(1996). very plesiomorphicmembers which share a few peculiarly advanced traits. These apomorphies coupled with the overall "primitiveness" was what mislead scientists as to their actual relationships. elongated shells; plastron reduced or hinged; carnivorous; bottom walkers
-musk glands on underside; barbelson the chin
Sternotherus odoratusstinkpot/ musk turtle( family kinosternon) (Fig 9)
The Pleurodira are identified by the method with which they withdraw their heads into their shells. In these turtles,
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Karyological analysis of order testudines 2016 the neck is bent in the horizontal plane, drawing the head into a space in front of one of the front legs. A larger overhang of the carapace helps to protect the neck, which remains partially exposed after retraction. This differs from the method employed by a cryptodiran, which tucks its headandneckbetween its forelegs, within the shell. .( Pearse,et al 1947)
.
(Fig 10)
Family chelidea
Like all pleurodirous turtles, the chelids withdraw their necks sideways into their shells, differing from cryptodires that fold their necks in the vertical plane. Frank Grützner;2006 They are all highly aquatic species with webbed feet and the capacity to stay submerged for long periods of timeThe highly aquatic nature of the group is typified by the presence of cloacal breathing in some species (Iverson,etal, 2012) are largely strike-and-gape hunters or foragers feeding on fish,
Matamata turtle (Fig11)
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Pelomedusidae is a family of freshwater turtles native to sub-Saharan Africa, with a single species, Pelomedusa subrufa, also found inYemen. They range in size from 12 to 45 cm (4.7 to 17.7 in) in carapace length, and are generally roundish in shape. They are unable to fully withdraw their heads into their shells, instead drawing them to the side and folding them beneath the upper edge of their shells, hence are called African side-necked turtles. (W. E.Rainey. 1980)
Pleomedusa subrufa turtle (Fig12)
The Podocnemididae are a family of turtles native to Madagascar and northern South America. They are side-necked turtles(Pleurodira), which means they do not retract their heads( Clark; 1967) backwards, but hide them sideways. These turtles are all aquatic, inhabiting streams and other flowing water. Their shells are streamlined to aid in swimming.
Podocnemis turtle (Fig13)
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(Fig 14)
In the nucleus of each cell, the DNA molecule is packaged into thread- like structures Called Chromosomes.Each chromosome is made up of DNA tightly coiled manytimes around proteins called histones that support its structure. (Gorman, G. C. 1973)Chromosomes are not visible in the cell’s nucleus—not evenunder a microscope—when the cell is not dividing. However, the DNA that makes up chromosomes becomes more tightly packed during cell division and is then visible under a microscope. Most of what researchers know about chromosomes was learned by observing chromosomes during cell division.Each chromosome has a constriction point called the centromere, which divides the chromosome into two sections, or ―arms.‖ The short arm of the chromosome is labeled the ―p arm.‖ The long arm of the chromosome is labeled the ―q arm.‖ The location of the centromere on each chromosome gives the chromosome its characteristic shape, and can be used to help describe the location of specific genes
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(Fig 15) what are type of chromosome?..
Metacentric Chromosomes Metacentric chromosomes have the centromere in the center, such that both sections are of equal length. Human chromosome 1 and 3 are metacentric. Submetacentric Chromosomes Submetacentric chromosomes have the centromere slightly offset from the center leading to a slight asymmetry in the length of the two sections. Human chromosomes 4 through 12 are submetacentric.
Acrocentric Chromosomes Acrocentric chromosomes have a centromere which is severely offset from the center leading to one very long and one very short section. Human chromosomes 13,15, 21, and 22 are acrocentric.
Telocentric Chromosomes Telocentric chromosomes have the centromere at the very end of the chromosome. Humans do not possess telocentric chromosomes but they are found in other species such as mice.
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(Fig 16)
A karyotype is a picture of all the chromosomes from an individual’s cells. A karyotype is a test used to check forchromosome abnormalities. A picture of a person’s chromosomes is created by staining the chromosomes with a special dye, photographing them through a microscope and arranging them in pairs. A karyotype gives information about the number of chromosomes a person has, the structure of their chromosomes and the sex of the individual
(Fig 17) What bands of chromosome ?......
Q-Banding
Quinacrine mustard, an alkylating agent, was the first chemical to band chromosomes viewed under a fluorescence microscope. Quinacrine dihydrochloride has subsequently been substituted by quinacrine mustard. The alternating bands of bright and dull fluorescence are
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Karyological analysis of order testudines 2016 called Q bands. The bright bands are primary composed of DNA rich in adenine and thymine, while the dull bands are rich in guanine and cytosine.
Q bands are especially useful for distinguishing the human Y chromosome and various chromosome polymorphisms involving satellites and centromeres of specific chromosomes.
G-banding
Giemsa has become the most commonly used stain in human cytogenetic analysis. Unlike Q-banding, G-banding usually requires pre- treating chromosomes with either salt or a proteolytic (protein- digesting) enzyme. When chromosomes are pre-treated with the proteolytic enzyme trypsin the process is called GTG banding. Giemsa stains preferentially regions rich in adenine and thymine. Therefore, G bands correspond closely to Q bands.Standard G band staining techniques allow between 400 and 600 bands to be seen on metaphase chromosomes. With high resolution G-banding techniques, as many as two thousand different bands have been catalogued on the twenty-four human chromosomes.
R-banding Reverse banding (R-banding) involves the incubation of slides containing metaphase chromosomes in hot phosphate buffer and stained with Giemsa. The banding pattern that results is essentially the reverse of G bands. R bands are GC-rich. The AT-rich regions are selectively denatured by heat leaving the GC-rich regions intact. Fluorochromes that are GC specific also produce a reverse chromosome banding pattern. R-banding is helpful for analyzing the structure of chromosome ends, since these areas usually stain light with G-banding.
C-Banding stains areas of heterochromatin, which is tightly packed and repetitive DNA. C-banding is specifically useful in humans to stain the centromeric chromosome regions and other regions containing constitutive heterochromatin - secondary constrictions of human chromosomes 1, 9, 16, and the distal segment of the Y chromosome long arm.
NOR-banding
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NOR-banding involves silver staining (silver nitrate solution) of the "nucleolar organizing region", which contains rRNA genes.
T-Banding
T-banding involves the staining of telomeric regions of chromosomes using either Giemsa or acridine orange after controlled thermal denaturation. T bands apparently represent a subset of the R bands because they are smaller that the corresponding R bands and are more strictly telomeric.
(Fig 18)
The nucleolus organizer region (NOR) or nucleolar organizer is a chromosomal region around which the nucleolus forms. This region is the particular part of a chromosome that is associated with a nucleolus after the nucleus divides. The region contains several tandem copies of ribosomal DNA genes. In humans, the NOR contains genes for 5.8S, 18S, and 28S rRNA clustered on the short arms of chromosomes 13, 14, 15, 21 and 22 (the acrocentric chromosomes).Nucleolus organizer regions (NORs) are head-to-tail arrays of genes encoding the precursor of the three largest ribosomal RNAs (18S, 5.8S and 25S in plants). NORs include active rRNA genes, which give rise to secondary constrictions of metaphase chromosomes, and silent rRNA genes, which are often highly compacted in dense heterochromatin.
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At metaphase, a proteinaceous remnant of the nucleolus often remains associated with the secondary constriction. Each rRNA gene at a NOR is nearly identical in sequence, although variation in size due to differences in the number of repeated DNA elements in the intergenic spacer region is common.
In karyotype analysis, a silver stain can be used to identify the NOR. Silver nitrate inserts into the NOR-associated protein in the stalks and satellites, staining the proteins dark black. The amount of stain deposited and the number of NORs differs among the population, although the cell should normally have a maximum of 10 NORs per cell.
INTRODUCTION of karyological analysis of testudines
Cytogenetics is the branch of genetics that studies the structure and behavior of chromosomes and their relation to human disease and disease processes. During the past three decades, the importance of clinical cytogenetics to the practice of obstetrics and gynecology has dramatically increased because clinical cytogenetics has a direct effect on the diagnosis, management, and prevention of many disorders that are caused by chromosome aberrations. For many chromosome disorders, physicians face medicolegal responsibilities in the areas of counseling, screening, and diagnosis, and obstetricians and gynecologists therefore must have knowledge about the human chromosome constitution and be able to apply basic principles of chromosome behavior to clinical practice. This chapter reviews
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Karyological analysis of order testudines 2016 important concepts and developments in cytogenetics and highlights their applications in the practice of obstetrics and gynecology.
OVER the past 10 years knowledge of turtle karyology has grown to such an extent that the order Testudines is one of the better known groups of lower vertebrates (Bickham,1983). Nondifferentially stained karyotypes areknown for 55% of cryptodiran turtle species and banded karyotypes for approximately 25%(Bickham, 1981). From this body of knowledge,as well as a consideration of the morphologicalvariation in the order, we herein present a general review of the cryptodiran karyological literature and a discussion of the evolutionary relationships of the higher categories ofcryptodiran turtles. Although this paper focuses on the Cryptodira (the largest suborder ofturtles), the Pleurodira also has been well studied in terms of standard karyotypes (Ayres etal., 1969; Gorman, 1973; Bull and etal, 1980)and a few have been studied with banding Cope recognized the currently widely acceptedsubordersCryptodira and Pleurodira.Two major differences between these two suborders are in the plane of retraction of the neckand the relationship between the shell and pelvic girdle. In the cryptodires ("hidden-necked"turtles), the neck is withdrawn into the body ina vertical plane and the pelvis is not fused toeither the plastron or carapace, whereas in the pleurodires ("side-necked" turtles) the pelvicgirdle is fused to both the plastron and carapaceand the neck is folded back against the body ina horizontal plane. Cope's suborder Athecae includes only the Dermochelyidae and is nolonger recognized. Most authors include theDermochelyidae among the Cryptodira (Gaffney, 1975a; Mlynarski, 1976; Wermuth and etal, 1977; Pritchard,1979). The families of the suborder Cryptodira arearranged in various superfamilies by several authors. The Testudinoidea, Chelonioidea
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Karyological analysis of order testudines 2016 andTrionychoidea are superfamilies common tomost of the recent classifications (Williams, 1950;Romer, 1966;Gaffney, 1975a; Mlynarski, 1976).However, the limits of these taxa are not uniformly agreed upon.The non-trionychoid freshwater and land cryptodiran turtles include the Chelydridae,Kinosternidae, Dermatemydidae, Platysternidae,Emydidae and Testudinidae and are usually placed in the Testudinoidea (Williams, 1950; Romer, 1966). Gaffney (1975a) includes the Kinosternidae and Dermatemydidae in theTrionychoidea. Mlynarski (1976) includes onlythe Emydidae and Testudinidae in the Testudinoidea.He recognizes the superfamily Chelydroidea to include the Chelydridae, Dermatemydidae, Kinosternidae and Platysternidae.The Chelonioidea includes the Cheloniidaeand the Dermochelyidae (Baur, 1893; Gaffney,1975a). Williams (1950), Romer (1966), andMlynarski (1976) recognize a separate superfamily,the Dermochelyoidea, for the familyDermochelyidae, and include only the Cheloniidae in the Chelonioidea. Most of the currently utilized family orsubfamily level taxa have been commonly recognized since Boulenger (1889). However, thereis no complete agreement regarding the levelat which certain taxa should be recognized. Parsons (1968) reviewed this confusing situationwith regard to the Chelydridae, Staurotypidae,Kinosternidae, Platysternidae, Emydidae and Testudinidae, as recognized here. Not mentioned by him are the inclusion of Platysternonin the Chelydridae (Agassiz, 1857; Gaffney,1975b) and the recognition of the Staurotypidae (Baur, 1891, 1893; Chkhkvadze, 1970). The above discussion of the history of cryptodiran taxonomy serves to illustrate the complexity of the relationships of the inclusive taxa.The taxonomic confusion seems to result from: 1) extensive convergent evolution in certainmorphological traits,
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2) the failure of someworkers to distinguish between shared primitive and shared derived character states
3)the lack of a widely accepted phylogeny of turtles. Chromosomal data are used in this paperin an attempt to solve some of the evolutionaryand classificatory problems. Cytogenetic information seems useful at this level because of thehigh degree of conservatism expressed in cheloniankaryotypes (Bickham, 1981). Additionally, the application of chromosome bandingtechniques solves one of the most troublesomeproblems in phylogeny reconstruction; namely,the determination of homologous characters.When two chromosomes have identical bandingpatterns it can safely be concluded that they are homologous. It is sometimes difficult to determine homology among morphological characters. For example, determination of homologies among the plastral scales of various turtle families is difficult. The fact that a scale is in the same position in members of different familiesdoes not necessarily imply homology (Hutchison et al, 1981
Most karyotypes were prepared by an in vivo technique as follows:
Blood was removed from the animal by means of a capillary tube inserted into the posterior angleof the eyelids, and pushed posteriorly into an orbital blood sinus. This was assumed to promote an immune response involving cell division in the spleen. An intraperitoneal injection of 0 1% colchicine was administered, at a dosage rate of 0.05 ml per gram body weight. The animal was killed 5 h after the colchicine injection. The spleen, and in males a testis, was removed, chopped finely and placedin 0.9% sodium citrate solution for 10 min. Air- dried smears were prepared by the techniques ofBaker et al. (1971),
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Karyological analysis of order testudines 2016 being fixed in 3 : 1 methanol : acetic acid. The smears were stained in a 4% aqueoussolution of Gurr's improved Giemsa stain for 10- 15 min, and mounted in canada balsam after beingdried for at least 1 h on a hotplate at 40-45OC.Some early results were obtained by a method similar to that of Baker et al. (1971). Bone marrowfrom the femur was flushed out with 0 9% sodium citrate solution, after the animal had been bled and treated with phytohaemagglutinin for 2 days. Results from this method were poor, with the exception of those for Physignathus lesueurii, and the method described above was subsequentlydeveloped. At least 10 divisions were examined from each preparation. The clearer of these preparations were photographed under oil immersion. The photomicrographs were printed and the lengths ofchromosomes measured with dial calipers used in a stepwise fashion. The total length of each chromosome arm was recorded, and the percentage of total macrochromosome length and centromeric indices were calculated for each division. Where possible somatic preparations were used for analysis
(Fig 18)
Chromosomes were arranged, according to themethod of Bickham (1975), into three groups(A:B:C:) where group A includedmetacentricsubmetacentricmacrochromosomes, groupBsubtelocentric-telocentric macrochromosomes,and group C
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Karyological analysis of order testudines 2016 microchromosomes. The A:B:C:formula is given after the diploidnumber inFig. 19 and in the text.This paper represents a synthesis and reanalysisof (mostly) published data. In reanalyzing the data we employed cladistic methodology(Hennig, 1966) in which sister groups were established by the determination of groups thatpossessed shared derived characters (synapomorphies).Because banded karyotypes were not available for the most appropriate outgroup taxon (Suborder Pleurodira: Family Chelidae)we employed an "internal" method of characterpolarity determination. Specifically, charactersthat were shared among families considered tobe distantly related, known from the fossil record to be early derivatives of the cryptodiraradiation, or thought to be morphologicallyprimitive, were considered as primitive (plesiomorphic)chromosomal characters. Because ofthe nature of karyotypic variation in cryptodiresthe analysis was rather straightforward. For example, dermatemydids are among the mostprimitive living turtles and their fossil historyextends back to the Cretaceous, as does the cheloniids which are thought to be an early offshootof the cryptodiran line. These two families possess species with apparently identical karyotypes. It is highly unlikely that these two familiespossess a synapomorphy at this level of the phylogeny. This would mean that these two familieswere more closely related to each other than toany other families studied, an arrangement thatappeared to conflict with every other line ofevidence in the literature. We therefore considered this karyotype to be primitive, at leastfor the non-trionychoid families, and the karyotypes of other families were derived from this
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(see below)
(Fig 19)
Fig. 19. G-band karyotype of a batagurine emydid (Chinemys reevesi, 2n = 52). The chromosomes are arranged into group A (metacentric or submetacentric macrochromosomes), group B (telocentric and subtelocentric macrochromosomes), and group C (microchromosomes)
Results and Discussion
The following discussion is segmented intothe commonly accepted family groups. In general, we have accepted each of the families asdistinct entities and do not question their validity . Emydidae
The two subfamilies of emydid turtles are characterized by different karyotypes.The predominantly New World emydines have2n = 50 and the predominantly Old World batagurinesmostly have 2n = 52 (Table 1). A fewbatagurine species also possess 2n = 50 (Table1), including Siebenrockiella crassicollis, the onlyemydid known to possess sex chromosomes (Carrand ,etal 1981). Bickham and etal (1976a)concluded that the primitive karyotype of theEmydidae was 2n = 52 and identical to that ofSacalia bealei and other Old World batagurines.This has been supported by recent findings that some testudinids have banded karyotypes identical to those of Chinemys reevesi and other batagurines
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(Dowler and etal, 1982). Fig. 19illustrates the karyotype of a batagurine (Chinemys reevesi) that possesses the proposed primitive emydid karyotype.The origin of the 2n = 50 emydine is unclear (Bickham and etal, 1976a). There is no karyotypic evidence to indicate emydinesare at all closely related to Rhinoclemmys, the only New World batagurine genus (Carr, 1981).There may be some hint of the batagurine-emydine transition in the finding of several speciesof Asiatic batagurines with 2n = 50 (Table1). Any relationship of the emydines to the 2n =50 batagurines will require evidence from othercharacter systems in order to establish its existence.
Testudinidae.
The karyology of this family isnot as well studied as that of the Emydidae but it seems certain that the primitive karyotype is2n = 52. Some species are known to possess Gbandpatterns identical to those of certain batagurines including Geochelone pardalis, G. elongate and G. elephantopus (Dowler and etal,1982). C-band variation exists among species ofGeochelone, and the karyotypes of Gopherusspecies differ from Geochelone species by themorphology and location of the nucleolar organizing region (NOR) (Dowler and etal,1982). Although this family is nearly world-wide in distribution and morhpologically diverse, theavailable data indicate a high degree of karyologicalconservatism.
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Table 1
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table 2
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Table 3
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Table 4
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(Table 5)
Platysternidae.
The standard karyotype of thesingle species of platysternid (Platysternon megacephalum) has 2n = 54 (Haiduk and Bickham,1982). This species appears to have close affinities to the Emydidae but is karyotypically distinct from all emydids thus far studied. BecauseP. megacephalum and emydids do apparently havesynapomorphic chromosomes that are notshared with chelydrids, Haiduk and Bickham (1982) considered P. megacephalum to comprisea family distinct from the Chelydridae (sensuGaffney, 1975b) and resurrected the Platysternidae (Gray, 1870), a move also suggested byWhetstone (1978).
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Karyological analysis of order testudines 2016
Staurotypidae.
This group is usually considered to be a subfamily (Staurotypinae) of theKinosternidae. Standard karyotypes of all three species in this group are known (Table 1; seeespecially Bull et al., 1974). The two species of Staurotypus are distinctive in possessing an XX/XY sex chromosome system (Bull et al., 1974;Sites et al., 1979a). Claudius angustatus, likenearly all other turtle species studied, does not possess heteromorphic sex chromosomes but appears to be otherwise karyotypically identicalto Staurotypus (Bull et al., 1974). Sites et al.(1979a, b) report banded karyotypes of 5. Salvini and show that this species possesses abiarmed second group B macrochromosome that appears to be homologous to an identicalelement in emydids and testudinids (and platysternidsbased on standard chromosome morphology). This chromosome is acrocentricinchelydrids,kinosternids, dermatemydids andcheloniids (Fig. 20). We conclude that the biarmedcondition is derived. Centric fusion of the ancestral acrocentric macrochromosome with amicrochromosome accounts for the presence of a subtelocentric macrochromosome in the common ancester of the Emydidae, Testudinidae,Platysternidae and Staurotypidae. This is indicative of the staurotypids belonging to a cladethat does not include kinosternids (Kinosternonand Sternotherus). This seems irreconcilable with
(Fig 20)
Fig. 20. G-band patterns of the second group B chromosomes of (left to right) a staurotypid, an emydid,a kinosternid
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Karyological analysis of order testudines 2016 and a cheloniid. The long arms ofall 4 taxa are identical; the short arms of the staurotypid and emydid are euchromatic and identical, however, the short arms of the kinosternid and thecheloniid are small and heterochromatic; see text forfurther discussion Chelydridae.
The two extant species of thisfamily have been studied for both standard (Table 1) and banded karyotypes (Haiduk and et al,1982). Chelydra serpentina and Macroclemystemminckii both have 2n = 52 but differ in themorphology of certain chromosomes. Haidukand et al (1982) conclude that these two species do not share any derived chromosomalcharacteristics with each other or with any other families of Cryptodira. However, the karyotypeof M. temminckii could be derived from that of C. serpentina. The latter is considered theprimitive karyotype for the family.
Kinosternidae
This family is comprised of twogenera and about 18 species and has been well studied karyotypically (Table 1). Early, and apparently inaccurate, reports aside (Table 1), allspecies thus far examined appear to possess2n = 56. Banded karyotypes (Bickham and etal, 1979; Sites et al., 1979b) indicate all speciespossess a large, subtelocentric macrochromosome not found in any other group of turtles.Kinosternids do not share any derived chromosomal characters with any other turtle family, including the staurotypids with which they
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Karyological analysis of order testudines 2016 are usually considered confamilial. An interesting variation was found in this family by Sitesetal. (1979b). Heterochromatin that stains darkin both G- and C-band preparations was found in Sternotherus minor, Kinosternon baurii and K.subrubrum, but not found in K. scorpioides. Thepresence of this type of heterochromatin wasconsidered to be a derived character (it is not found in closely related families) shared amongthe three species that possess it, indicating that the genus Sternotherus has affinities with temperate species of Kinosternon.
Dermatemydidae.
The single extant speciesofthis family (Dermatemys mawii) possesses 2n = 56
(Table 1). There are no uniquely derived elements and this species shares no derived chromosomes with any other family.
Cheloniidae.
Members of this family possess2n = 56 (Table 1). Banding data indicate cheloniidsand dermatemydids are karyotypicallyindistinguishable (Bickham et al., 1980; Carr etal., 1981). Early reports of other diploid numbers and sex chromosomes have not been substantiated by recent studies using current techniques.
Trionychidae.
Members of both subfamilies(Cyclanorbinae and Trionychinae) have 2n =66 (Table 1). Reports of other diploid numbershave been unsubstantiated in subsequent studies. The report of 2n = 52-54 in Trionyx leithii(Singh et al., 1970) was due to the misidentification
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Karyological analysis of order testudines 2016 of this specimen (Kachuga dhongoka, Emydidae;Singh, 1972). The 2n = 66 karotype wasconsidered by Bickham et al. (1983) to be theprimitive karyotype for the family. Banding comparisons between Trionyx and Chelonia revealed little homology between the Trionychidae and Cheloniidae (Bickham et al., 1983).
Carettochelyidae The single extant species(Carettochelys insculpta) has 2n = 68 (Bickham et al., 1983). Although no banding data have beenreported for this species, the standard karyotype is very similar to the 2n = 66 karyotype of trionychids.
Taxonomy The acceptability of using karyotypicdata in order to draw phylogenetic inferences and erect a classification at the level offamily and higher is based upon the conservatism of the karyotypic character system. Bycharacter system, we refer to a suite of characters and character states which may be presumed to be closely enough related to be withinthe realm of influence of the same set ofevolutionary constraints. According to this line ofresasoning then, karyotypic data constitute acharacter system separate from the charactersystems associated with electrophoretic data orcranial osteology, etc. The level at which characters are relatively constant within a group isthe point at which thosecharacters are of systematic utility and those characters are said tobeconservative (Farris, 1966). Our studies anda review of the pertinentliteratureindicate thatfamily level groups within the Cryptodira arecharacteristically karyotypically homogeneousand that the significant variation (in the phylogenetic sense) is observable interfamilially. Itis upon these premises that we propose the classification in Table 6 based upon our cladisticanalysis of the karyotypic data.This classification is conservative in that allfamilies commonly recognized are maintained,even though in two instances
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Karyological analysis of order testudines 2016 there are familypairs which we cannot karyotypically distinguish [i.e., Cheloniidae-Dermatemydidae
Table 6
(Fig 21)
Fig 21. Cladogram showing the hypothesized relationships of the higher categories of cryptodiranturtles. The diploid number and the number of chromosome pairs in groups A:B:C (Fig. 19) in the proposedprimitive karyotype of each family (and both subfamilies of Emydidae) are shown. Because the trionychoidfamilies are so divergent, the A:B:C formulas are notgiven (Bickham et al., 1983). Characters 1 -5 are listed and discussed in the text. to recognize the Staurotypinae as a separate
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Karyological analysis of order testudines 2016 family, the Staurotypidae. This conclusion is incongruentwith data from other character systems. Many morphological studies report similarities between the Kinosternidae andStaurotypidae (among these Williams, 1950;Parsons, 1968; Zug, 1971). Most such studieshave not attempted cladistic analyses (two exceptions are Gaffney, 1975; et al, 1981). There seems no obvious orsimple manner in which to reconcile the conflicting data from the karyotypic character system and the overwhelming amount of data fromvarious morphological character systems. Inrecognizing the Staurotypidae, we have madeexplicit our prediction of its relationships toother testudinoid families. Independent confirmation or refutation of these relationships will determine the merit of this move.The three superfamilies are all considered tobe holophyletic. Fig. 21 presents a cladogram that we believe best reflects the branching sequence of the evolution of this group. The Testudinoidea and Chelonioidea may be sistergroups but this is as yet unproved. The primitive karyotypes of these two taxa are identical,2n = 56 (character 1 in Fig. 21), and very different from that of the Trionychoidea, 2n = 66-68 (character 2 in Fig. 21),but we do not yet know the polarity of these character states(Bickham et al., 1983). All testudinoid and chelonioid turtles possessat least seven group A macrochromosomes(character 1 in Fig. 21). Among the testudinoidfamilies, aclade that includes Staurotypidae,Platysternidae, Testudinidae, and Emydidae canbe identified by the presence of a biarmed second group B macrochromosome (character 3in Fig. 21; Fig. 20). Another clade includes thePlatysternidae, Testudinidae and Emydidae allof which primitively possess nine group A macrochromosomes (Fig. 1; character 4 in Fig. 21).A clade including the Emydidae and Testudinidae is characterized by a 2n = 52 9:5:12 primitive karyotype (Fig. 19; character
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Karyological analysis of order testudines 2016
5 in Fig. 21).Species of the emydid subfamily Emydinae allpossess a karyotype derived from the primitive9:5:12 arrangement (Bickham and etal,1976a). The Dermatemydidae, Kinosternidae andChelyridae possess no chromosomal synapomorphiesand the branching sequence of these families is not obvious from chromosomal, morphological or serological data. However, the Chelydridae is usually considered to be mostclosely related to the Emydidae (McDowell,1964; Zug, 1971;Frair, 1972; Haiduk and etal, 1982) and the dermatemydids, morphologically one of the most primitive families ofturtles, are considered closely allied to the Kinosternidae (Zug, 1971; Frair, 1972; Gaffney,1975b).The Cheloniidae and Dermochelyidae areconsidered to comprise the suborder Chelonioidea.There are no karyotypic data availablefor Dermochelys coriacea so the relationship between this species and cheloniids has yet to betested chromosomally. But, these two families are closely related morphologically and serologically(Frair, 1979). We follow most other workers in giving this group full superfamilialstatus, recognizing that they have invaded anadaptive zone, the marine environment, that is distinctly different from that of most other turtles. It must be emphasized that Chelonia mydas(Chelonioidea) and Dermatemys mawii (Testudinoidea) appear karyotypically identical and weinterpret this to be the primitive karyotype of these two superfamilies.The superfamily Trionychoidea includes onlythe Trionychidae and Carettochelyidae. Thesetwo taxa are closely related chromosomally as well as morphologically and their karyotypesare distinctly different from those of species of the other two superfamilies. Some workers haveincluded the Kinosternidae andDermatemydidae in the Trionychoidea (Gaffney, 1975a).The chromosomal data do not support such anarrangement because of the disparity in diploid number and chromosome morphology between testudinoids (including kinosternids and dermatemydids) and trionychoids (Bickhametal.,1983Historical review of taxonomic relationships.—The primary subdivisions of the order
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Karyological analysis of order testudines 2016 comprisingthe turtles have undergone a great many namechanges and rearrangements over the last 100years. Cope (1871) presented an arrangementof the families into suborders which is still widely accepted today. Until Cope, the subordinal andsuprafamilial classification of turtles was primarily based on differences in the digits amongthe sea turtles, the aquatic turtles and/or theterrestrial tortoises. Hoffman (1890) and Kuhn(1967) present reviews of the early classifications.Cope recognized the currently widely accepted suborders Cryptodira and Pleurodira.Two major differences between these two suborders are in the plane of retraction of the neck).and the relationship between the shell and pelvic girdle. In the cryptodires ("hidden-necked"turtles), the neck is withdrawn into the body ina vertical plane and the pelvis is not fused toeither the plastron or carapace, whereas in the pleurodires ("side-necked" turtles) the pelvicgirdle is fused to both the plastron and carapaceand the neck is folded back against the body in a horizontal plane. Cope's suborder Athecaeincludes only the Dermochelyidaeand is nolonger recognized. Most authors includetheDermochelyidae among the Cryptodira (Gaffney,1975a; Mlynarski, 1976; Wermuth andMertens, 1977; Pritchard, 1979).A few authors recognize the Trionychoidea(sensu Siebenrock, 1909) and/or the Chelonioidea(sensu Baur, 1893) at a suprafamilialrank equivalent with the Cryptodira and Pleurodira (Boulenger, 1889;Lindholm, 1929; Mertens et al., 1934). The suborder Cryptodira isused here in the sense of Williams (1950) and subsequent authors and includes all living nonpleurodiranturtles.The families of the suborder Cryptodira arearranged in various superfamilies by several authors. The Testudinoidea, Chelonioidea andTrionychoidea are superfamilies common tomost of the recentclassifications (Williams, 1950;Romer, 1966;Gaffney, 1975a; Mlynarski, 1976).However, the limits of these taxa are not uniformly agreed upon.The non-trionychoid freshwater and landcryptodiran turtles include the Chelydridae,Kinosternidae, Dermatemydidae, Platysternidae,Emydidae and
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Karyological analysis of order testudines 2016
Testudinidae and are usually placed in the Testudinoidea (Williams, 1950;Romer, 1966). Gaffney (1975a) includes the Kinosternidae and Dermatemydidae in the© 1983 by the American Society of Ichthyologists and HerpetologistsTrionychoidea. Mlynarski (1976) includes onlythe Emydidae and Testudinidae in the Testudinoidea. He recognizes the superfamily Chelydroideato include the Chelydridae, Dermatemydidae, Kinosternidae and Platysternidae.The Chelonioidea includes the Cheloniidae and the Dermochelyidae (Baur, 1893; Gaffney,1975a). Williams (1950), Romer (1966), and Mlynarski (1976) recognize a separate superfamily,the Dermochelyoidea, for the familyDermochelyidae, and include only the Cheloniidae in the Chelonioidea.The Trionychoidea usually includes both theTrionychidae and Carettochelyidae(Mlynarski,1976), but Williams (1950) and Romer (1966)recognize the Carettochelyidae separately intheCarettochelyoidea.Most of the currently utilized family orsubfamily level taxa have been commonly recognized since Boulenger (1889). However, thereis no completeagreementregarding the level at which certain taxa should be recognized. Parsons (1968) reviewed this confusing situationwith regard to the Chelydridae, Staurotypidae ,Kinosternidae, Platysternidae, Emydidae and Testudinidae, as recognized here. Not mentioned by him are the inclusion of Platysternonin the Chelydridae (Agassiz, 1857; Gaffney, 1975b) and the recognition of the Staurotypidae (Baur, 1891, 1893;Chkhkvadze, 1970).The above discussion of the history of cryptodirantaxonomy serves to illustrate the complexity of the relationships of the inclusive taxa.The taxonomic confusion seems to result from: 1) extensive convergent evolution in certainmorphological traits , 2) the failure of someworkers to distinguish between shared primitive and shared derived character states and
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Karyological analysis of order testudines 2016
3)the lack of a widely accepted phylogeny of turtles. Chromosomal data are used in this paperin an attempt to solve some of the evolutionaryand classificatory problems. Cytogenetic information seems useful at this level because of the high degree of conservatism expressed in chelonian karyotypes (Bickham, 1981). Additionally, the application of chromosome banding techniques solves one of the most troublesomeproblems in phylogenyre construction; namely,the determination of homologous characters.When two chromosomes have identical bandingpatterns it can safely be concluded that they arehomologous. It is sometimes difficult to determine homology among morphological characters. For example, determination of homologiesamong the plastral scales of various turtle families is difficult. The fact that a scale is in thesame position in members of different familiesdoes not necessarily imply homology (Hutchison and Bramble, 1981).
(Fig 22)
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Karyological analysis of order testudines 2016
Testudindae 1) Karyotype ofGeochelone denticulate , male 2n =52 , with an 2:6:12 complement of groups A:B:C 2) karyotype of Geochelone carboaria , male 2n =52, 9:5:12 (bickham and et al 1976b)
(Fig 23) emydidae 3) karyotype of chryemys terrapin , male 2n =50 8:5:12 4) karyotype of chrysemys decorate, male 2n=50 , 8:5:12
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Karyological analysis of order testudines 2016
5) karyotype of chrysemys stejnegeri vicina ,male , 2n =50, 8:5:12(Bickham and BAKER 1976b)
(Fig 24) Batagurinae 6)Karyotype of rhinoclemys pulcherrima ,female 2n= 52 , 6:5:15 7) karyotype of rhinoclemys puctularia female 2n=56 (BICKHAM AND BAKERb)
(Fig25)
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Karyological analysis of order testudines 2016
kinostrenidae 8) karyotype of kinosternon scorpioides , female 2n = 56, 7:6:15 (Bickham an et al 1976b)
(Fig 26) dermatemydidae
FlG. 26.—Standard karyotype of Dermatemys mawii with chromosomes arranged into groups: (A) metacentric to submetacentric macrochromosomes, (B) telocentric to subtelocentric macrochromosomes, and (C) microchromosomes. The standard karyotype of D. mawii(2n = 56) is presented in Figure 26. Chromosomes are arranged according to Bickham(1975) into group A metacentric orsubmetacentric macrochromosomes,group B telocentric or subtelocentric macrochromosomes, and group C microchromosomes.There are 7, 5, and 16 pairs of chromosomes in groups A, B, and C, respectively. A heteromorphic pair ofsex chromosomes is not present in themale specimen examined.
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Karyological analysis of order testudines 2016
(Fig 27)
Fig. 27 G- and C-banded metaphase chromosome karyotypes of female (a, c) andmale (b, d) A. spinifera, respectively. e Enlarged images of the highly heterochromatic microchromosomes in A. spinifera, indicating theWin female (Fem) and the heterochromatic microchromosome pair (m) in both females and males. A large block of the female-specific chromosome (W) is Giemsa faint and
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Karyological analysis of order testudines 2016
C-positive. The arrow indicates the female-specific chromosomes with large C-positive block. Note that the Z is morphologically indistinguishable from several other microchromosomes with similar banding pattern. Scale bar=10 μm
Trionychidae
1972; BICKHAM ET AL. 1983). NINE PAIRS OF MACROCHROMOSOMES AND 24
PAIRS OF MICROCHROMOSOMES WEREIDENTIFIED, DIFFERING SLIGHTLY FROM THE
REPORT BYBICKHAM ET AL. (1983) OF EIGHT PAIRS OF MACROCHROMOSOMES.
THE MACROCHROMOSOMES IDENTIFIED HEREINCLUDED TWO PAIRS OF
METACENTRIC, FOUR PAIRS OF SUBMETACENTRIC,AND THREE PAIRS OF
ACROCENTRIC CHROMOSOMES(FIG. 27). THE CENTROMERE POSITION OF THE 24
PAIRS OFMICROCHROMOSOMES COULD NOT BE DETECTED ACCURATELYDUE TO
THEIR SMALL SIZE, WHICH IMPEDES THE UNAMBIGUOUSPAIRING OF SOME
MICROCHROMOSOMES WITH THEIR GBANDED
Fig 28 Chelonoidea With the method described above, metaphases were identified with a good distribution and number, allowing the identification of sets of chromosomes. The
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Karyological analysis of order testudines 2016 non-banded mitotic chromosomes were visualized by Giemsa staining. Chelonoidis carbonaria revealed a diploid number of 2n = 52 chromosomes, in both sexes, divided into three groups (A, B, C). Group A was composed of 28 chromosomes (3 metacentric pairs, one acrocentric and 10 submetacentric pairs), group B consisted of seven pairs of acrocentric chromosomes, and group C showed five pairs of microchromosomes (Figure 1A,B). Sex chromosomes were not observed.
SUBORDER PLEURODIRA INTRODUCTION of pleurodira
Turtles of the suborder Pleurodira are divided into two families, the Chelidae and the Pelomedusidae, which are clearly separated by both morphological (Gaffney, 1977) and molecular (Shaffer et al., 1997) features. The Chelidae consists of nine genera, five of which are found in Australia and New Guinea and four in South America (Ernst and Barbour, 1989). Conflicting phylogenies have been proposed for the Chelidae, but recent phylogenetic analysis based on molecular markers (Seddon et al., 1997; Fujita et al., 2004) support the monophyly of the Australian/New Guinea and South American chelid turtles. The chelid genus Hydromedusa (commonly known as snake-necked turtles) consists of two species of semi-aquatic turtles that have an extremely long throat: H. maximiliani, restricted to the southeast region of Brazil; and et al, distributed throughout southern and southeastern Brazil, northeastern Argentina, Uruguay and southeastern Paraguay.
The chromosomes of birds, fishes and some reptile groups are highly variable in terms of size and morphology, and are characterize by bimodal or asymmetric karyotypes composed of macro and
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Karyological analysis of order testudines 2016 microchromosomes. Turtle karyotypes show two general tendencies based on the presence or absence of microchromosomes but there is much variation between groups. For example, the chromosome number in the order Chelonia ranges from 2n = 26 in Podocnemis dumeriliana (Ayres et al., 1969) to 2n = 96 in Platemys platycephala (Bull and Legler, 1980; Bickham et al., 1985). Also, while karyotypic studies have frequently been published for turtles from the suborder Cryptodira, information about Pleurodires is scarce and fragmented and mainly based on conventional staining techniques.
In this paper describe the almost complete karyotypic characterization of Hydromedusa tectifera using several staining techniques and in situ Fluorescence Hybridization (FISH).
SUPERFAMILY PELOMEDUSIDAE
Pelomedusids have low diploid numbers and few microchromosomes (2n = 26–36); the five largest chromosomes are homologous in the three genera.
The big-headed side-neck river turtle, Peltocephalusdumerilianus (Schweigger, 1812), occurs in the Amazonregion and belongs to the superfamily Pelomedusoides (approximately24 living species), which comprises the families Pelomedusidae, with two living genera: Pelomedusaand Pelusios
represented by one, and at least 15 species, respectively;and Podocnemididae, with three living genera:the monotypic Erymnochelys and Peltocephalus, andPodocnemis comprising six species (Ayres et al., 1969;Vitt and et al, 2009). In Podocnemididae cytogeneticdata are scarce and based mostly on conventional staining.The Podocnemis and Erymnochelys species (P. erythrocephala,P. expansa, P. lewyana, P. sextuberculata, P.unifilis, P. vogli and E. madagascariensis) present a diploidnumber (2n) of 28, with a karyotype composed of
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Karyological analysis of order testudines 2016 fivemacrochromosomes (M) and nine microchromosomes (m)(Ayres et al., 1969; Huang and Fred Clark 1969; Rhodin etal., 1978; Bull and Legler, 1980; Fantin andet al, 2011;Gunski et al., 2013). The exception is Peltocephalus dumerilianus that presents 2n = 26 with 4 M and9m, the lowest diploid number in Testudines (ranging from 2n = 26 to 2n = 96) (Ayres et al., 1969; Bull and et al,1980). The available cytogenetic data for this species report a karyotype that is similar to those of other Podocnemididae,in which differentiated sex chromosomes are absentand a conspicuous secondary constriction is observed the karyotype of P.dumerillianus was characterized for the first time using routine differential techniques, such as GTG, CBGbandingand Ag- NOR staining (Seabright, 1971; Sumner,For all individuals, at least 20 metaphases were analyzedfor determining the 2n = 26 and FN = 52 karyotype,as described by Ayres et al., 1969, with a conspicuous secondary constriction on pair 1 (Figure 1A). GTG-banding patterns allowed the identification and the pairing of allchromosomes (Figure 1B). CBG-bands were tenuous at thepericentromeric region of most pairs, except for pair1
(Fig 29)
Figure 29- Karyotype (A and B) and metaphases (C-E) of Peltocephalus dumerilianus, 2n = 26 and FN = 52. (A) Conventional staining. Inset, pair
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Karyological analysis of order testudines 2016
1 from other metaphase showing the conspicuous secondary constriction. (B) GTG-banding pattern. (C) CBG-banding pattern. Note the conspicuous C-positive bands on pair 1. Inset, pair 1 bearing positive Ag-NORs. probes. Positive signals (green) are seen at the termini of all chromosomes. (E) Mapping of 45S rDNA. Positive FISH signals (red) are at the secondary constrictionregion of pair 1.
FAMILT PODOcnemis
To determine karyotypes we counted 35 cellsin each individuals from each species. Our resultsshowed that the karyotypic number forboth species of Podocnemis is 2n = 28 chromosomes,consisting of 5 pairs of macrochromosomesand 9 pairs of microchromosomes, with the following morphologies: 16m + 2sm + 10aand NF = 46 (Figure 1). Silver-nitrate staining All species of the genusPodocnemis present a chromosomal number of2n=28, which is extremely low compared withkaryotypes described for other species. Few karyotypic studies have been conductedon the genus Podocnemis (AYRES et al. 1969;RHODIN et al. 1978; BULL and et al1980; ORTIZet al. 2005), the work by Ayres et al. (1969)is the only one that presents cytogenetic studies for P. expansa and P. sextuberculata. However,there are still no studies on chromosomal evolutionwithin the family Podocnemidae, nor arethere any comparative studies of chromosomal banding.This work describes the karyotypes and thelocalization of the nucleolar organizer regions(NORs) in two species of the genus Podocnemis
(Fig 30)
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Karyological analysis of order testudines 2016
Fig. 30 — Karyotype of Podocnemis sextuberculata (above)microchromosomes and P. expansa with2n = 28.
Family chelidea
Chelids have high diploid numbers and many microchromosomes (2n = 50–64) and are similar in this respect to cryptodires (2n = 50–66).
(Fig 31)
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Karyological analysis of order testudines 2016
Fig 32
The chromosome complement of all our Hydromedusa tectifera specimens was 2n = 58, of which 22 were macrochromosomes and 36 microchromosomes (Figure 1a 33). It was possible to precisely determine the position of the centromere in the macrochromosomes, and we observed one submetacentric chromosome pair, one metacentric pair and nine pairs of acrocentric chromosomes, giving a total of 62 chromosome arms. No sex chromosome heteromorphism was observed. This diploid number agrees with the study of Bull and Legler (1980),
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Karyological analysis of order testudines 2016
(Fig 33)
(Fig34)
The G-banding permitted the visualization, especially in the macrochromosomes, of a pattern of bands that enabled better identification and pairing of the chromosomes as well as the construction of an ideogram Figure 34). Such a pattern is similar, but not identical, to that observed in other Pleurodiran turtles, due to the presence and absence of some bands when compared to the patterns found by Bull and Legler (1980) in Pelomedusoid turtles (a group related to the Chelidae). This variation in the G-banding pattern in Pleurodiran turtles establishes a different karyotypic evolution from that identified for the suborder Cryptodira. Previous reports have suggested genomic stability in Cryptodiran turtles, in which both the banded chromosome
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Karyological analysis of order testudines 2016 morphology (Bickham, 1981) and the DNA sequences inside the chromosomes (Muhlmann-Díaz et al., 2001) remain unchanged for millions of years.
WE CONCLUDE, FOR TWOREASONS, THAT THE PRIMITIVE KARYOTYPE OF THE
SUBORDER CRYPTODIRA IS MOST LIKELY THE 2N = 56KARYOTYPE OF CHELONIID AND
DERMATEMYDID TURTLES. FIRST, THESE ARE AMONG THE MOST ANCIENTFAMILIES IN
THE SUBORDER (BOTH DATE FROM THECRETACEOUS), AND SECOND, THIS KARYOTYPE
IS HIGHLY GENERALIZED AND COULD HAVE GIVEN RISE TO THEDIVERSITY OF
KARYOTYPES IN THE SUBORDER BY A MINIMUM NUMBER OF EVENTS. A PRIMITIVE
KARYOTYPEMORE SIMILAR TO THAT OF TRIONYCHOID TURTLES (2N =66-68) CANNOT
ENTIRELY BE RULED OUT (BICKHAMET AL., 1983). COMPARISONS WITH KARYOTYPES
OFTHE SPECIES OF PLEURODIRA DO NOT SOLVE THE PROBLEM BECAUSE SPECIES OF
THE CHELIDAE ARE KNOWNTO POSSESS DIPLOID NUMBERS IN THE 2N = 56 RANGEAS
WELL AS THE 2N = 66 RANGE (BULL AND LEGLER,
1980). HOWEVER, THE PRIMITIVE KARYOTYPE OF THEPLEURODIRA WAS
CONSIDERED BY Bull and Legler (1980) to be 2n = 50-54 which is consistent withour hypothesis of a 2n = 56 ancestral karyotypefor the Cryptodira.If the above hypothesis is true, then chromosomal evolution in the Trionychoidea involved an increase in the diploid number by a reduction in the number of macrochromosomesand an increase in the number of microchromosomes.However, chromosomal evolution inthe Testudinoidea reduced the diploid numberby an increase in the number of macrochromosomes and reduction of the number of microchromosomes. Bickham and Baker (1979) note that specieswithin a family or subfamily possess identical or very similar karyotypes. However, karyotypic comparisons among families and subfamilies
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Karyological analysis of order testudines 2016 almost always reveal variation. A more refinedanalysis of the pattern of karyotypic variationin turtles (Bickham, 1981) suggests that the rateof karyotypic evolution has decelerated and thatMesozoic turtles evolved at a rate twice as fastas their descendants. Additionally, the kinds ofchromosomal rearrangements incorporatedduring the diversification of cryptodiran families differ from the kinds of rearrangements incorporated during the evolution of modernspecieThe above described pattern of karyotypicevolution is consistent with thecanalizationmodel of chromosomal evolution (Bickham andBaker, 1979). Under this model, evolution ofthe karyotype is driven by natural selection because the chromosomal rearrangements altergenetic regulatory systems. Changes that areadaptive accumulate more rapidly during theearly radiation of a lineage. As time goes onmore and more adaptive linkage groups areproduced. Further chromosomal rearrangement tends to break up adaptive gene sequencesand the rate of chromosomal evolution slowsdown. Thus, in an ancient group such as turtles,the process of canalization has had such a longperiod of time to act that karyotypic evolutionamong modern forms is virtually nonexistent.However, when karyotypic comparisons aremade of taxa that diverged early during turtleevolution, such as comparisons of the primitivekaryotypes of families, variation is found to bemore pronounced.Models that explain karyotypic evolution bypopulation demography, such as deme size, donot apply to turtles. The classical model of chromosomal speciation (White, 1978) requires fixation of chromosomal rearrangements in smalldemes due to genetic drift orinbreeding. Thereis some question as to whether chromosomal speciation is in fact a viable process (Bickhamand Baker, 1979, 1980; Futuyma and Mayer,1980), but even if it is, it certainly is not operative in
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Karyological analysis of order testudines 2016 turtles. There are no known chromosomal races in turtles. This could be explainedby turtles characteristically not having smallpopulation sizes or other demographic factorsthat promote the fixation of chromosomal rearrangements by genetic drift or inbreeding.However, turtles display such a diversity of demographic characteristics (Auffenberg andIverson, 1979; Bury, 1979; Bustard, 1979) thatthis explanation seems untenable. Turtles exhibit a diverse array of morphological types and occur in nearly all habitatsavailable to reptiles. Some, such as the migratory sea turtles, are highly vagile but others,such as tortoises, have relatively low vagility.Reproductive rates also vary. The green turtlemay lay as many as 200 eggs in a single clutch,some emydids may lay only a single large egg.While there are certainly many species thatcharacteristically have large population sizes, wecan point to many that probably do not. For example, kinosternids and emydids that occurin the arid western United States and Mexicos. often are found in isolated stock tanks, ponds,intermittent streams and permanent springs.Population sizes are often small and there isprobably very little migration among populations.Many of the above mentioned biological characteristics of turtles conceivably could promotechromosomal speciation. That it does not occurin a major radiation (Cryptodira) does not meanthat the process is not viable in other taxa, butits absence is somewhat unexpected. In conclusion, population parameters are poorly correlated with chromosomal variability in turtles andin principle we agree with the criticisms of thechromosomal speciation models espoused by Bickham and et al (1979, 1980) and Futuyma and et al (1980).
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