Genetic diversity studies in Lithuania

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Genetic diversity studies in Lithuania
Workshop Bioplatform in Lithuania launch of
biodiversity research forum 2004 10 15 , Vilnius

• Algimantas Paulauskas
• Vytautas Magnus university

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Topic of Genetic diversity studies

• Taxonomy differentiation of sibling species
• Ecology
• test-object of genotoxical impurity (small
  rodents)
• genetic variability in different biotopes
• Differentiation of reintroduced species and
  populiation
• Gene pool estimation of rare and endangered
  species
• Biodiversity of plant and animal genetic
  resources

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Material and Methods Study Area

• Samples were obtained during research expeditions
  in different region of the Palaearctic
  (Lithuania, Kamchatka, Chukotka, Lena, Ob and
  Kara rivers).
• Tissue and blood samples (400-500 ?l) were
  collected in heparin tubes and frozen at -20?C
  till use.

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Methods

• Citogenetic
• karyological
• micronucleus
• sperm heads anomalies
• Morphological studies
• non-metric and metric parameters
• dominant lethal mutation
• Allozymes
• DNA
• PCR, RAPD, Microsatelit
• Sequences

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Objects

• Plants
• Paris VU
• trees LFRI
• Animals
• Parasites Borrelia sp., protists and
  helminth LVA, VMU, VUEI
• Molluscs VMU, VU, VUEI
• Insects VU
• Fish VMU, VU, VUEI
• Birds VMU, VU, VUEI
• Mammals VMU, VU, VUEI
• Domestic animals LVA, LAI, VMU, VUEI

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Analysed species
Microtus arvalis
Apodemus uralensis
Apodemus agrarius
Microtus agrestis
Castor fiber
Microtus rossiaemeridionalis
Micromys minutus
Apodemus flavicollis
Clethrionomys glareolus
Microtus oeconomus
Castor fiber
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Systematics
Karyotype differentiation of Microtus arvalis and
M. rossiaemeridionalis R. Maeikyte,V.Morkunas
1999
Helminth J. Bariene, R.Petkeviciute,
G.Staneviciute
Insects J. Puplesiene
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DNA RAPD (primer ROTH-180-06, elektroforeze 2
agaroses gelyje). 1, 18 Gene RulerTM 100bp DNA
Lader Plus 2, 3 M. rossiaemeridionalis 4, 5
M. arvalis 6, 7 M. agrestis 8, 9 M.
oeconomus 10 M. minutus 11, 12 C.
glareolus 13, 14 A. agrarius 15, 16 A.
flavicollis 17 A. uralensis.
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Genetic similarity among rodents species based on
RAPD data (general ).
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Dendrogram showing genetic relations among
rodents species based on RAPD data (ROTH-180-01)
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Average Cluster Analysis of the GENETIC
VARIATION OF MIGRATING GEESE
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Rodents - test-object of genotoxical impurity 1
Cletrionomys glareolusMicrotus arvalisM.
agrestisApodemus sylvaticusA. agrarius
chromosomu aberacijos dominuojancios letalios
mutacijos (embrionu mirtingumas) spermatozoidu
galvuciu anomalijos
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Rodents - test-object of genotoxical impurity 2
Genetinis kintamumas agropramoniuose rajonuose ir
draustiniuose
R.Lekevicius 1973-2000 LietuvaV.Morkunas
1980-2004 Lietuva S. Juzolenas 1976
KretuonasA. Paulauskas 1983 Maeikiai H.
Balandis 1983 IgnalinaV. Juknevicius 1985 J
onavaA. Vilkas 1986 MaeikiaiD. Mateikaite
1988 ElektrenaiG. Vyniauskis 1991
Kedainiai
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Differentiation of reintroduced species and
populiation

• 1 Fenetiniai (kraniologija) Ulevicius A.
  1988-20042 Baltymu (izofermentu) analize
  Paulauskas A., Ulevicius A. 1998-20043 DNR
  polimorfizmas Paulauskas A., Norvaiaite
  B. Ulevicius A. 2002-2004

Europinio bebro reintrodukcijos vietos
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Similarity of C. fiber populations 82 beavers
(1999-2002)

• Merkys
• Sesupe

• Minija
• Sventoji

• Dubysa
• Nemuno delta

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Genetic variability of population
Rodents Cletrionomys glareolus, Microtus
arvalis M. agrestis Apodemus sylvaticus A.
agrarius Beavers Castor fiber Mole Talpa
europaea
1 Baltymu (izofermentu) analize 2 DNR
polimorfizmas Paulauskas A., Skiriute D.,
Jukeviciute E., Maeikyte R., Balciauskas L.
1998-2004 Paulauskas A., Norvaiaite B.
Ulevicius A. 1998-2004 Paulauskas A.,
Starodubaite M., Sruoga A. 2001-2004
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Genetic variability at 12 loci in Lithuanian
populations of European Bevers
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Genetic variability at 12 loci in Lithuanian
populations of European Bevers
Similarity 0,80 0,83 0,87 0,90
0,93 0,97 1,00 ----------------
--------------------------------

--------------- MERKYS
------------

--------------- EUPE -------------------
--------- -------------- VENTOJI
-------------

-------------- MINIJA
------------------------
DUBYSA --------------------------------

------------------------ NEMUNO DELTA
----------------------------------------
-------- 0,80 0,83 0,87 0,90
0,93 0,97 1,00  Distance 0,60
0,50 0,40 0,30 0,20 0,10
0,00 ----------------------------------
--------------
------------------------ MERKYS
---

------------------------ EUPE
----------- ----------------------
VENTOJI
-----
----------------------
MINIJA
-------------------------------- DUBYSA
-------

-------------------------------- NEMUNO DELTA
----------------------------------------
-------- 0,60 0,50 0,40 0,30
0,20 0,10 0,00
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(No Transcript)
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• The biggest genetic distances followed by Nei
  (1972) are between Kaunas site and Zarasai island
  populations (D3.059 to 3.244)
• The biggest genetic identity are between islands
  Berzai and Bebrai (D0.948). Arkliai island was
  formed before 50 years only.

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Apodemus agrarius
Distance .40 .33 .27 .20
.13 .07 .00 ------------------
------------------------------

SAKIAI


KEDAINIAI


BIRZAI ---------------------------
--------------------- .40 .33
.27 .20 .13 .07 .00
Similarity .90
.92 .93 .95 .97 .98
1.00 ----------------------------------
--------------

SAKIAI 01LT


KEDAINIAI 0


BIRZAI 0 ----------------
-------------------------------- .90
.92 .93 .95 .97 .98
1.00
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APODEMUS FLAVICOLLIS Similarity .90
.92 .93 .95 .97 .98
1.00 ----------------------------------
--------------

SAKIAI


VIESVILE

KURSIU
NERIJA

PAGRAMANTIS


VARNIAI


KEDAINIAI


KAUNO MARIOS


VISTYTIS


BIRZAI ---------------
--------------------------------- .90
.92 .93 .95 .97 .98
1.00
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Dendrogram showing genetic relations among
Apodemus flavicollis population based on RAPD data
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Population genetic analysis of host-parasite
coevolution

• The studies have revealed that different
  population and species of both the parasites and
  the hosts may have different genetic polymorphism
  and heterozygosity.
• We studied the population genetic variation of
  Trematodes N. attenuatus and N. ephemera
  (multiple-host parasite with low host
  specificity) and they intermediate (molluscs) and
  definitive hosts (birds, rodents) .
• Population dynamic studies which consider genetic
  polymorphism of helminths and hosts have shown
  fluctuations of different genotypes in space and
  time.

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Analysis of host-parasite coevolution (1980-2003)

• Parasite Trematodes N. attenuatus (6
  population) and N. ephemera (5 population)

• Intermediate hosts freshwater molluscs Limnaea
  stagnalis (6 population) and Planorbarius corneus
  (4 population)

• Definitive hosts dabbling ducks (563 samples of
  Anas acuta, A. clypeata, A. crecca, A.
  querquedula, A. falcata, A. penelope, A.
  platyrhynchos, from 11 population), 43
  individuals of migrating White-fronted geese
  (Anser albifrons).

• Facultative hosts rodents Microtus oeconomus, M.
  arvalis, M. agrestis (12 population) and
  Clethrionomys glareolus (7 population)

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genetic variation of parasite population

• The investigation of genetic variation within and
  between Lithuania population of Trematodes N.
  attenuatus and N. ephemera (multiple-host
  parasite with low host specificity) was showed
  that levels of genetic variation had different
  values in different population and different
  time ranged from H0.171 to H0.353 for
  N.attenuatus (1-5) and from H0.251 to H0.427
  for N. ephemera (6-10) .

Genetic variability in Trematodes N. attenuatus
and N. ephemera populations (standard errors in
parentheses)

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genetic variation of parasite population

• Parasite strains which are adapted to their local
  host population are more virulent to introduced
  host from another populations which previously
  had no experience with this parasite strain.
• In experiment the infections rate the
  establishment of N.attenuatus (6-10) and N.
  ephemera (6-10) in duklings and goslings with
  different genotypes was different.

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genetic variation of intermediate hosts population

• On the other hand the genetic polymorphism among
  host species and population have been also
  different in space and time for intermediate
  host (molluscs) ranged from H0.0 (Arion sp.) to
  H0.63 (Cerithium caerulleum), in different
  population of Limnaea stagnalis ranged from
  H0.28 to H0.46, in population of Planorbarius
  corneus ranged from H0.18 to H0.38.

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genetic variation of definitive hosts population

• Genetic variation among geographically different
  populations in the Palaearctic region (Lithuania,
  Kamchatka, Lena, Ob and Kara rivers) of
  definitive host - dabbling ducks (563 samples of
  Anas acuta, A. clypeata, A. crecca, A.
  querquedula, A. falcata, A. penelope, A.
  platyrhynchos, investigated in 1980-1998) show
  that this rate fluctuated among geographically
  different populations
• for A. acuta between Ho0.264 from the Ob river
  and Ho0.331 from Kamchatka
• for A. penelope between Ho0.185 from the Ob and
  Ho0.263 from the Lena river,
• for A. platyrhynchos sub-populations between
  Ho0.242 (Vente 1998, Lithuania) and Ho0.366
  (Antanava 1987, Lithuania).

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Intraspecific genetic variability of
White-fronted geese

• Three flocks of Anser albifrons were
  investigated

Migratory flyways of White-fronted goose (Anser
albifrons) Map according to Scott and Rose,
1996.
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What is future of Genetic diversity studies in
Lithuania ?
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Thank you for attention !