Zbornik - Društvo genetičara Srbije

IV-Pos-12 ZBORNIK ABSTRAKATA III KONGRESA GENETIÈARA SRBIJE 129
Subotica, 30. novembar - 4. decembar 2004.
VARIJABILNOST AKTIVNOSTI PEROKSIDAZA I SADRAJA
TEŠKIH METALA U ÈETINAMA LINIJA POLUSRODNIKA CRNOG BORA
(Pinus nigra Arn.)
V. Isajev 1 , Ksenija Radotiæ 2 i M. Duèiæ 1
1
Šumarski fakultet, Beograd
2
Centar za multidisciplinarne studije, Univerzitet u Beogradu, Beograd
Ekstrakcija enzima je raðena najpre homogenizacijom u 5 mL 0.1 M TRIS-HCl puferu
pH 7.6 koji sadri 1 mM DTT i 1 mM EDTA na ledenom kupatilu po proceduri Mocquot
et al. (1996) i Weckx i Clijsters (1996)Koncentracija proteina je uraðena po metodi Bradford
(Bradford, 1976). Za odreðivanje koncentracije metala u èetinama crnog bora,
uzorci su pripremani prema proceduri za «suvo spaljivanje» Uzorak je oèitavan na
apsorberu Varian AA-10. Detekcioni limit za Cd je od 0.02 do 3 g/mL, za Cu je 0.03-10
g/mL i za Ni 0.1-20 g/mL. U cilju utvrðivanja koliki je uticaj genetièke raznolikosti
linija polusrodnika na jedan od kljuènih enzima ukljuèenih u odgovor na stres, i
zaduenog za proces lignifikacije u æelijskom zidu, praæena je aktivnost enzima
peroksidaza. Takoðe je praæena aktivnost ovog enzima u zavisnosti od kiselosti
medijuma. Izoelektrofokusiranjem utvrðeni su izoenzimi koji su prisutni u èetinama
analiziranih osam linija crnog bora.
Rezultati ovog rada pokazuju prisustvo izraenog varijabiliteta u koncentraciji ukupnih
proteina, sadraju teških metala i aktivnosti enzima peroksidaza kod analiziranih linija
polusrodnika crnog bora. Ovo govori da proteini imaju razlièit nivo ekspresije u pojedinim
linijama crnog bora. Takoðe enzim peroksidaza pokazuje razlièitu aktivnost koja
zavisi od genetske konstitucije stabala iz analiziranih linija polusrodnika, jer se, na
osnovu velièine plantae i šeme sadnje biljaka, moe smatrati da je uticaj spoljašnjih
faktora sveden na minimum.
VARIABILITY OF PEROXIDASE ACTIVITY AND HEAVY METAL
CONTENT IN THE NEEDLES OF AUSTRIAN PINE (Pinus nigra Arn.)
HALF-SIB LINES
Enzyme extraction was performed first by homogenisation in 5 mL 0.1 M TRIS-HCl
buffer pH 7.6 containing 1 mM DTT and 1 mM EDTA in ice bath, by procedure
Mocquot et al. (1996) and Weckx and Clijsters (1996). The concentration of proteins was
determined by Bradford (Bradford, 1976) method. For the measurement of metal concentration
in Austrian pine needles, the samples were prepared by the procedure for «dry
burning». The sample was read off on the absorber Varian AA-10. Detection limit for Cd
is from 0.02 to 3 g/mL, for Cu 0.03-10 g/mL and for Ni 0.1-20 g/mL. The enzyme
activity of peroxidases was monitored in order to determine the effect of genetic variability
of half-sib lines on one of the key enzymes included in the response to stress, and responsible
for the process of lignification in the cell wall. Also, the activity of this enzyme
was monitored depending on the medium acidity. The isoenzymes present in the needles
of the analysed eight Austrian pine lines were determined by isoelectric focusing.
The results of this study show the pronounced variability of the concentration of total
proteins, heavy metal concentration and enzyme activity of peroxidases in the study Austrian
pine half-sib lines. This infers that proteins have different levels of expression in individual
Austrian pine lines. Also the enzyme of peroxidases shows different activity depending
on the genetic constitution of the trees of the analysed half-sib lines, because
based on he seed orchard size and the planting pattern, it can be assumed that the effect of
external factors is reduced to a minimum.