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Cloning low
temperature responsive genes from Pachysandra terminalis
Suping
Zhou, Roger Sauve, Abdullah Abdullah, Sarah Bhatti and Deborah Long
Institute of Agricultural and Environmental Research
Seminar Series
Tennessee State University, Nashville, TN
September 9, 2004
Pachysandra terminalis is a cold hardy, evergreen plant species. In
order to identify genes that are regulated by low temperature
fluctuation, Pachysandra terminalis were subjected to 4 and 0 C
low temperature treatment. Low temperature induced genes were identified
from treated plants using cDNA differential display. The cDNA fragments
were cloned onto PCR-trap vectors. Low temperature regulation of these
genes was confirmed by reverse-northern blot. Two hundred fifty clones
were selected out being up-regulated by low temperature treatments.
Sequence analyses have identified that these genes can be classified
into three groups, stress-related, photosystem-related and unidentified.
To further study the regulation of these genes by temperature
fluctuation, the plants were treated at 4 C and 0 C and then incubated
at 25 C. Some of them were subjected to 0 C treatment again after
recovery at 25 C for 24 h. Northern blot analysis of the leaf RNA from
the treated leaves identified few genes that are up-regulated by low
temperature treatments. These genes were down regulated during warming
up at 25 C. Further treatment at 0 C failed to induce their high
expression.
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