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