RNA Differential Display of Heat Shock Protein Genes in Wheat
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RNA Differential Display of Heat Shock Protein Genes in Wheat Samah M. Eldemery1*, El-Sayed A. El-Absawy2, Amal A. Abdelaziz1 1 Molecular Biology Department, GEBRI, Sadat City, Minoufiya University, Egypt 2 Bioinformatics Department, GEBRI, Sadat City, Minoufiya University, Egypt *Corresponding author: [email protected]2012 Climate change and abiotic stress adversely affect agriculture and crop production. Three 10-mer arbitrary primers were used to produce RNA fingerprinting of wheat genotypes under heat stress and three specific heat shock protein gene primer pairs to identification different members of heat shock protein multi gene families in wheat using differential display technique. Four different hexaploid wheat varieties were used: two Egyptian varieties (‘Giza 168’ and ‘Gemmiza 7’), one Mexican heat tolerant variety (‘SERI M 82’), and one Chinese heat susceptible variety (‘Chinese Spring’). 10-day-old seedlings were exposed to heat stress at 37°C for 2 hours, and the leaf tissue was used for cDNA synthesis. The random primers generated a number of amplified expressed genes ranging from 5 for the OPW03+DDPT primers combination to 15 for the OPC07+DDPC primers combination. Comparable number of amplified fragments (amplified expressed genes) was obtained for different gene primers pairs (e.g. hsp16.9, hsp70, and hsp101 primers pairs). Both random and specific primers were able to differentiate between the heat treatment and control treatment by amplifying specific fragments. The cluster analysis of differential display data separated the genotypes into two main clusters. One contains the heat treated and untreated types of ‘Giza168’ and ‘SERI M 82’ varieties along with the untreated type of the ‘Gemmiza 7’ variety. The second cluster included both types of the ‘Chinese Spring’ variety (heat treated and untreated) along with the heat treated ‘Gemmiza 7’ variety. The results suggest that both ‘Gemmiza 7’ and ‘Chinese Spring’ varieties are heat susceptible, while ‘Giza 168’ and ‘SERI M 82’ are heat tolerant varieties. The results revealed that differential display was successful for gene expression profile comparison in different thermo tolerance wheat varieties under heat shock condition. Key words: Wheat, differential display, HSP genes, heat shock response ــ ــRNA differential display pattern of 12 random primers combination on wheat control and heat shocked samples. Dendrogram of Differential display patterns of the total RNA from control and heat shocked wheat seedlings RNA differential display pattern of 12 specific primers combination on wheat control and heat shocked samples.
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Transcript of RNA Differential Display of Heat Shock Protein Genes in Wheat
RNA Differential Display of Heat Shock Protein Genes in Wheat Samah M. Eldemery1*, El-Sayed A. El-Absawy2, Amal A. Abdelaziz1
1 Molecular Biology Department, GEBRI, Sadat City, Minoufiya University, Egypt
2 Bioinformatics Department, GEBRI, Sadat City, Minoufiya University, Egypt *Corresponding author: [email protected]
Climate change and abiotic stress adversely affect agriculture and crop production. Three 10-mer arbitrary primers were used to produce
RNA fingerprinting of wheat genotypes under heat stress and three specific heat shock protein gene primer pairs to identification different
members of heat shock protein multi gene families in wheat using differential display technique. Four different hexaploid wheat varieties
were used: two Egyptian varieties (‘Giza 168’ and ‘Gemmiza 7’), one Mexican heat tolerant variety (‘SERI M 82’), and one Chinese heat
susceptible variety (‘Chinese Spring’). 10-day-old seedlings were exposed to heat stress at 37°C for 2 hours, and the leaf tissue was used for
cDNA synthesis. The random primers generated a number of amplified expressed genes ranging from 5 for the OPW03+DDPT primers
combination to 15 for the OPC07+DDPC primers combination. Comparable number of amplified fragments (amplified expressed genes) was
obtained for different gene primers pairs (e.g. hsp16.9, hsp70, and hsp101 primers pairs). Both random and specific primers were able to
differentiate between the heat treatment and control treatment by amplifying specific fragments. The cluster analysis of differential display
data separated the genotypes into two main clusters. One contains the heat treated and untreated types of ‘Giza168’ and ‘SERI M 82’
varieties along with the untreated type of the ‘Gemmiza 7’ variety. The second cluster included both types of the ‘Chinese Spring’ variety
(heat treated and untreated) along with the heat treated ‘Gemmiza 7’ variety. The results suggest that both ‘Gemmiza 7’ and ‘Chinese
Spring’ varieties are heat susceptible, while ‘Giza 168’ and ‘SERI M 82’ are heat tolerant varieties. The results revealed that differential
display was successful for gene expression profile comparison in different thermo tolerance wheat varieties under heat shock condition. Key words: Wheat, differential display, HSP genes, heat shock response
ــ ــ
RNA differential display pattern of 12
random primers combination on wheat
control and heat shocked samples.
Dendrogram of Differential display patterns
of the total RNA from control and heat
shocked wheat seedlings RNA differential display pattern of 12
specific primers combination on wheat
control and heat shocked samples.
