V o l u m e 53(1):55-76, 2009
Acta Biologica
Szegediensis
http://www.sci.u-szeged.hu/ABS
Mitochondrial sequence variation in ancient horses f r o m the Carpathian Basin
and possible m o d e r n relatives
Katalin Priskin
Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, Szeged, Hungary
W h a t e v e r the place, ethnic c h a n g e s a l w a y s leave their footprint in the local culture and genetic m a k e u p and the s a m e applies to the different t y p e s of h o r s e s m o v i n g with their o w n e r s . T h i s thesis is concerned with the mitochondrial control region g e n o t y p e s of ancient and
m o d e m h o r s e s f r o m the C a r p a t h i a n Basin, w h e r e in the late 9th cenury the incoming pagan H u n g a r i a n tribes p e r m a n e n t l y c h a n g e d the
population.
Studies of m i t o c h o n d r i a l D N A h a v e s h o w n that m o d e m h o r s e s are descended f r o m at least 77 different wild m a r e s , with a last c o m m o n
a n c e s t o r o v e r 3 0 0 , 0 0 0 years a g o . and s o probably inhabiting very different regions. Despite this matrilineal genetic diversity, correlations
b e t w e e n m o d e m b r e e d s of h o r s e s and mitochondrial g e n o t y p e are o f t e n u n c o m m o n . T h i s is p r o b a b l y b e c a u s e horse-trading and horsestealing. s o m e t i m e s o v e r long distances, h a v e been p o p u l a r and profitable for a long time.
T o d e t e r m i n e genetic diversity and origin of horse p o p u l a t i o n s in the Carpathian Basin at the time of the Avars and of the H u n g a r i a n
C o n q u e s t , m i t o c h o n d r i a l D N A a n a l y s i s w a s undertaken o n 31 archaeological horse r e m a i n s , e x c a v a t e d f r o m authentic, well-dated Avar
and p a g a n H u n g a r i a n burial sites. B a s e d o n a s u p p o s e d relationship, m o d e m Hucul and A k h a l Teke h o r s e s w e r e included in the analysis.
T o teveal r e l a t i o n s h i p s to odier a n c i e n t and recent b r e e d s . m t D N A s e q u e n c e s f r o m 7 9 b r e e d s representing 9 1 3 individual s p e c i m e n s w e r e
c o m b i n e d w i t h o u r s e q u e n c e data. S e q u e n c e s w e r e aligned and truncated to a length of 247 b p to a c c o m o d a t e p u b l i s h e d short s e q u e n c e s
( n u c l e o t i d e p o s i t i o n s 1 5 4 9 5 - 1 5 7 4 0 of r e f e r e n c e s e q u e n c e X 7 9 5 4 7 ) .
E s t i m a t i o n of s t a n d a r d diversity m e a s u r e s , s u c h as h a p l o t y p e diversity (h) and n u c l e o t i d e diversity (jt) w e r e p e r f o r m e d in D n a S P
4.50.2.
To c o m p a r e o u r s a m p l e s with other m o d e m a n d ancient horse sequences, 921 previously p u b l i s h e d e q u i n e m t D N A C R s e q u e n c e s with
f u l l y o v e r l a p p i n g s t a n d a r d 2 4 7 b p lengths were o b t a i n e d f r o m the database ( h t t p : / / w w w . n c b i . n l m . n i h . g o v / G e n b a n k ) .
M e d i a n - j o i n i n g n e t w o r k w a s c o n s t r u c t e d using the N E T W O R K 4.5.1 s o f t w a r e ( F l u x u s T e c h n o l o g y Ltd. at w w w . f l u x u s - e n g i n e e r i n g .
c o m ) to reveal a p p r o x i m a t e g e n e a l o g i c a l relationships a m o n g the h a p l o t y p e s f o u n d in o u r ancient and recent breeds and the h a p l o g r o u p indicating sequences.
B o t h g e n e t i c d i s t a n c e s a n d h a p l o l t y p e - b a s e d m e t h o d s indicate a clear separation b e t w e e n h o r s e s of the Avar and H u n g a r i a n leading
n o b l e s . Avar s e q u e n c e s w e r e genetically h e t e r o g e n e o u s , closely related to Eastern breeds; (with M o n g o l i a n and north Russian V y a t s k a y a
g r o u p s ) . T h i s A s i a n r e l a t i o n s h i p can also b e seen in the g e n e t i c distance matrix and the h a p l o t y p e n e t w o r k .
B y contrast, b e s i d e the great heterogenity and u n i q u e haplotypes. the ancient H u n g a r i a n h o r s e s s h o w e d a relatively c l o s e relationship
w i t h the T u r k o m a n A k h a l T e k e a n d N o r w e g i a n F t j o r d . It c o n f i r m s the a s s u m p t i o n of Hecker ( 1 9 5 5 ) a n d is an a d m i x t u r e to the clew u p of
the H u n g a r i a n origin. A f t e r a short c o m m u n i c a t i o n of B j a r n s t a d (2003), different d i s t a n c e m e a s u r e s suggest genetic associations between
n o r t h e r n E u r o p e a n horse b r e e d s and the M o n g o l i a n native horse. By all means, the high variability of H u n g a r i a n horse haplotypes may b e
c o n n e c t e d w i t h the w e l l - a t t e s t e d , c o n t i n e n t - w i d e raiding h a b i t s of the ancient H u n g a r i a n s . T h e H u c u l data s h o w n o such relationship. O u r
results s h o w that t h e e t h n i c c h a n g e s i n d u c e d by the H u n g a r i a n C o n q u e s t in the late
century w e r e a c c o m p a n i e d by a similar c h a n g e in
the s t a b l e s of the C a r p a t h i a n B a s i n .
Supervisor: Raskó István
e-mail: [email protected]
Isolation and characterization of bacterium and phage isolates which have
biocontrol ability against Pseudomonas strains pathogenic to Pleurotus
ostreatus
Enikő Sajben
D e p a r t m e n t of Microbiology, University of Szeged, Szeged Hungary
Pleurotus ostreatus is o n e of the m o s t extensively cultivated m u s h r o o m s in the world: h o w e v e r significant loss of c r o p and quality arises
f r o m b a c t e r i a l d i s e a s e s c a u s i n g b y d i f f e r e n t bacterial p a t h o g e n s . T h e yellowing of Pleurotus ostreatus and the b r o w n blotch disease of
Agaricus bisporus, c a u s e d b y Pseudomonas
tolaasii is well k n o w n ( L e e and C h a 1998). T h e b a c t e r i u m p r o d u c e s the toxins tolaasins that
disrupt the cellular m e m b r a n e by f o r m i n g p o r e s (Rainey et al. 1992).
Ps. tolaasii can be identify easily, w i t h lolaasin toxin g e n e specific primers. T h i s method is m u c h m o r e reliable than the white line
test. T h e Ps. tolaasii is a b l e to m a k e a p h e n o t y p i c switch; this variant f o r m is n o n p a t h o g e n i c and d i f f e r s f r o m the wild type in a r a n g e of
b i o c h e m i c a l a n d physiological characteristics (Cutri e t al. 1984).
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V o l u m e 53(1):55-76, 2009
Acta Biologica Szegediensis
http://www.sci.u-szeged.hu/ABS
M o r e o v e r o t h e r fluorescent p s e u d o m o n a d s such as Ps. agarici, Ps. constantinii
and Ps. gingeri can cause various s y m p t o m s as well.
T h e s a m e d e g r e e of discoloration m a y be c a u s e d by dissimilar species of p s e u d o m o n a d s . suggesting that the f a c t o r s are not e x c l u s i v e to a
particular p s e u d o m o n a d species ( G o d f r e y et al. 2 0 0 1 ) .
M a n y investigations have been carried out to find an appropriate method for preventing o r controlling this disease. T h e r e are trials to use
of c h e m i c a l w a s h formulations, including such c h e m i c a l s as calcium chloride, s o d i u m hypochlorite, hydrogen peroxide, b r o n o p o l and antibiotics in watering m u s h r o o m s , but n o n e of t h e m has been f o u n d to be fully effective and n o n - t o x i c to h u m a n s ( W o n g and P r e e c e 1985).
Biological control by competition has a l s o been investigated. Potential bacterial c o n t r o l a g e n t s h a v e been isolated a n d s h o w n to be
active antagonists (Nair and Fahy 1972).
In this study, the Pseudomonas
strains of an infected o y s t e r m u s h r o o m f a r m in H u n g a r y w e r e investigated. Sixty strains w e r e isolated
o n G o u l d ' s S I m e d i u m , which consistently gives high selectivity and good recoveries of fluorescent p s e u d o m o n a d s with s a m p l e s o b t a i n e d
f r o m a variety of habitats. SI m e d i u m has several a d v a n t a g e s over other media used for the isolation of fluorescent p s e u d o m o n a d s . T h e
identification of the strains w a s carried out by s e q u e n c i n g a part of the rpoB g e n e or in s o m e other c a s e s a part of the 16S r D N A g e n e . T h e
rpoB g e n e c o d e s f o r the R N A p o l y m e r a s e P-subunit. it is a highly conserved essential g e n e , so it c o u l d be used for bacterial identification.
T h e pathogenicities of the strains w e r e tested o n yeast extract media in Petri-dishes. in direct c o n f r o n t a t i o n tests.
To find and effective antagonistic agent, w e isolated and evaluated fluorescent p s e u d o m o n a d s . bacilli and lytic b a c t e r i o p h a g e s , against
Pseudomonas
tolaasii, and other pathogenic Pseudomonas
strains.
Cutri SS. Macaule BJ and Robert YWP (I984) Characteristics of pathogenic non-fluorescent (smooth) and non-pathogenic fluorescent (rough) forms of Pseudomonas
tolaasii and Pseudomonas gingeri. J Appl Bacteriol 51:291 -298.
Godfrey SAC. Harrow SA. Marshall JW and Klena. JD (2001) Characterization by 16S rRNA sequence analysis of pseudomonads causing blotch disease of cultivated
Agaricus bisporus. Appl Environ Microb 67:4316-4323.
Lee H-l and Cha J-S (1998) Cloning of a DNA fragment specific to Pseudomonas tolaasii causing bacterial brown blotch disease of oyster mushroom (Pleurotus
ostreatus). Kor J Plant Pathol 14:177-183.
Nair NG and Fahy. PC (1972) Bacteria antagonistic to Pseudomonas tolaasii and their control of brown blotch of the cultivated mushroom Agaricus bisporus. 1 Appl
Bacteriol 35:439-442.
Rainey PB. Brodcy CL and Johnstone K (1992) Biology of Pseudomonas tolaasii. cause of brown blotch disease of the cultivated mushroom. Adv Plant Pathol 8:95117.
Wong WC and Preece. TF (1985) Pseudomonas tolaasii on mushroom (Agaricus bisporus) crops: bactericidal effects of six disinfectants and their toxicity to mushrooms. J Appl Bacteriol 58:269-273.
Supervisor L£szl6 Manczinger
E-mail: [email protected]
Interacting protein partners o n Drosophila telomeres
Sándor Takács
D e p a r t m e n t of Genetics, University of Szeged, Szeged, Hungary
T h e proliferation disrupter (prod) gene in Drosophila e n c o d e s a 346- amino-acid protein that localizes strongly to the centric heterochromatin
of the s e c o n d and third c h r o m o s o m e s as well as to > 4 0 0 e u c h r o m a t i c sites, and all t e l o m e r e s . In Drosophila melanogaster.
three t e l o m e r i c
d o m a i n s can b e distinguished by D N A s e q u e n c e and by p r o t e i n s associated with t h e m : the end of the c h r o m o s o m a l D N A m o l e c u l e (capping c o m p l e x ) , the retrotransposon array consisting of three non-long terminal repeat r e t r o t r a n s p o s o n s , HeT-A. TART, and TAHRE ( H T T ) ,
and the s u b t e l o m e r i c repetitive t e l o m e r e associated s e q u e n c e s , repetitive region (TAS). C h r o m o s o m e length in Drosophila
is m a i n t a i n e d
by targeted transposition of the three telomere-specific n o n - l o n g terminal repeat r e t r o t r a n s p o s o n s . H T T . to the c h r o m o s o m e end.
I m m u n o f l u o r e s c e n c e stainings of different mutant t e l o m e r e s clearly s h o w e d , that Prod b i n d s to the HeT-A e l e m e n t of the H T T array.
We c o u l d a l s o s h o w that Prod binding represses HeT-A transcription, nevertheless d o e s not influence H T T length. Reduction of Prod levels
in h e t e r o z y g o u s prod"**10 null m u t a n t flies results in elevated levels of HeT-A transcripts in ovaries as well as in third instar larvae, w h i l e
it has no e f f e c t on g e n o m i c HeT-A c o p y n u m b e r , w h i c h w e used as a m e a s u r e m e n t for H T T length.
To identify p r o t e i n s that m a y interact with P r o d , we p e r f o r m e d a yeast t w o - h y b r i d screen, u s i n g the C l o n t e c h M a t c h m a k e r c D N A library and the entire prod c D N A as bait. A b o u t 120 0 0 0 c D N A c l o n e s w e r e tested and a b o u t 100 potential Prod interactors w e r e identified.
Prod w a s f o u n d 4 times, suggesting that Prod interacts with itself. We a l s o identified Z 4 as o n e of the interacting proteins, w h i c h is o n e
of the f e w p r o t e i n s k n o w n to b e associated with the H T T t e l o m e r i c d o m a i n . T h e C h r o m a t o r protein, w h i c h has previously been s h o w n to
c o - i m m u n o p r e c i p i t a t e with Z 4 and co-localize with Z 4 in i n t e r b a n d s of polytene c h r o m o s o m e s and at s o m e telomeres, was a l s o f o u n d to
interact with Prod. We could verify the latter interaction with c o - i m m u n o p r e c i p i t a t i o n e x p e r i m e n t .
Proteins U b a 2 and Lesswright w e r e the strongest interactors of Prod in the yeast t w o hybrid screen.Uba2 is the E l S U M O activating
e n z y m e regulating the initial steps of s u m o y l a t i o n . - a posttranslational protein m o d i f i c a t i o n s y s t e m m o d i f y i n g protein activity - while
the second step is p e r f o r m e d by the E2 u b i q u i t i n - c o n j u g a t i n g e n z y m e e n c o d e d by the lesswright g e n e in Drosophila. C o m p u t e r s e q u e n c e
analysis s h o w e d that Prod (and C h r o m a t o r ) have potential sumoylation sites. This raised the possibility that either Prod itself is s u m o y l a t e d
o r s u m o y l a t i o n e n z y m e s are recruited by Prod to s u m o y l a t e Prod-associated proteins. O u r i m m u n o s t a i n i n g s c o n f i r m this notion, d e m o n strating that most if not all sumoylated proteins o n t e l o m e r e s are located on the H I T array.
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