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Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.44 [2006] >
IJEB Vol.44(06) [June 2006] >
| Title: | Optimization of RQRT-PCR protocols to measure  -1,3-glucanase mRNA levels in infected tissues of rubber tree (Hevea brasiliensis) |
| Authors: | Thanseem, I
Thulaseedharan, A |
| Keywords: | Actin
-1,3-Glucanase
Hevea brasiliensis
Phytophthora
Relative RT-PCR |
| Issue Date: | Jun-2006 |
| Publisher: | CSIR |
| 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 -1,3-glucanase gene was studied in
the pathogen infected tissues of rubber. The -1,3-glucanase gene was found to be
induced in infected leaf tissues and reached a peak at 48 h after inoculation.
The -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. |
| Page(s): | 492-498 |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.44(06) [June 2006]
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