Optimization of RQRT-pCR protocols to measure beta-1,3-glucanase mRNA levels in infected tissues of rubber tree (Hevea brasiliensis).
Indian J Exp Biol
;
2006 Jun; 44(6): 492-8
Article
in English
| IMSEAR
| ID: sea-55813
ABSTRACT
RQRT-PCR technique was evaluated for its validity as an alternative to Northern blotting for quantification of plant gene expression in diseased tissues of Hevea. Reliable RT-PCR results could be obtained by co-amplification of housekeeping actin gene as the internal control along with the gene of interest. The product of interest was quantified relative to that of the internal control by measuring net intensity of bands. Expression levels of defense-related beta-1,3-glucanase gene was studied in the pathogen infected tissues of rubber. The beta-1,3-glucanase gene was found to be induced in infected leaf tissues and reached a peak at 48 h after inoculation. The beta-1,3-glucanase gene expression during pathogen infection was determined through Northern blot hybridization also, using 18S RNA as the internal control. RQRT-PCR and Northern hybridization showed almost similar results, thereby validating the use of this technique to study the gene expression in rubber.
Full text:
Available
Index:
IMSEAR (South-East Asia)
Main subject:
Phytophthora
/
Rubber
/
Autoradiography
/
Time Factors
/
RNA
/
RNA, Messenger
/
Blotting, Northern
/
Blotting, Southern
/
DNA, Complementary
/
Plant Leaves
Country/Region as subject:
South America
/
Brazil
Language:
English
Journal:
Indian J Exp Biol
Year:
2006
Type:
Article
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