Low expression profiles of heat stress-related genes inCapsicum annuum
- 168 Downloads
A cDNA library was constructed for hot pepper plants that had been heat-shock-treated. We used a modified differential screening method, double negative screening, to isolate 500 cDNA clones that represented genes with low expression levels under conditions of high-temperature stress. Of those 500 clones, 200 were randomly selected for single-read sequencing from the 5′ ends. After annotation with Blastx, the sequence was applied to InterProScan to scan for functional motifs of proteins. Among the cDNA clones analyzed, about 41% of the ESTs could not be functionally classified. However, of those that could be, the largest portion of the ESTs (15%) were assigned to the category of cell rescue and defense; genes involved in cell cycle/DNA processing constituted the smallest group, comprising 1% of the ESTs. Genes related to energy and protein fates constituted the second (10%) and third (9%) largest groups, respectively. Finally, 3% of the ESTs were assigned to transcription, and 2% to signal transduction. The high portion of unclassified ESTs probably resulted from the screening method, which was designed for low-expression messages. Likewise, the high number of ESTs for cell rescue and defense suggests that many genes with low levels of expression are associated with the stress response.
KeywordscDNA library double negative screening high-temperature stress hot pepper (Capsicum annuum)
Unable to display preview. Download preview PDF.
- Asamizu E, Nakamura Y, Sato S, Tabata S (2000) A large scale analysis of cDNA inArabidopsis thaliana: Generation of 12,028 non-redundant expressed sequence tags from normalized and size-selected cDNA libraries. DNARes 7: 175–180Google Scholar
- Basha E, Lee GJ, Breci LA, Hausrath AC, Buan NR, Giese KC, Vierling E (2004) The identity of proteins associated with a small heat shock potein during heat stressin vivo indicates that these chaperones protect a wide range of cellular functions. J Biol Chem 279: 7566–7575PubMedCrossRefGoogle Scholar
- Carninci P, Shibata Y, Hayatsu N, Sugahara Y, Shibata K, Itoh M, Konno H, Okazaki Y, Muramatsu M, Hayash-izaki Y (2000) Normalization and subtraction of cap-trapper-selected cDNAs to prepare full-length cDNA libraries for rapid discovery of new genes. Genome Res 10: 1617–1630PubMedCrossRefGoogle Scholar
- Chen W, Provart NJ, Glazebrook J, Katagiri F, Chang HS, Eulgem T, Mauch F, Luan SH, Zou G, Whitham SA, Budworth PR, Tao Y, Xie Z, Chen X, Lam S, Kreps JA, Harper JF, Si-Ammour A, Mauch-Mani B, Heinlein M, Kobayashi K, Hohn T, Dangl JL, Wang X, Zhu T (2002) Expression profile matrix ofArabidopsis transcription factor genes suggests their putative functions in response to environmental stresses. Plant Cell 14: 559–574PubMedCrossRefGoogle Scholar
- Falk S, Maxwell DP, Laudenbach DE, Huner NPA (1996) Photosynthetic adjustment to temperature.In NR Baker, ed, Photosynthesis and the Environment. Kluwer Academic, Dordrecht, pp 367–385Google Scholar
- Pellegrineschi A, Reynolds M, Pacheco M, Brito RM, Alm-eraya R, Yamaguchi-Peng Z, Staub JM, Serino G, Kwok SF, Kurepa J, Bruce BD, Vierstra RD, Wei N, Deng XW (2001) The cellular level of PR500, a proteir complex related to the 19S regulatory particle of the protea-some, is regulated in response to stresses in plants. Mol Biol Cell 12: 383–392Google Scholar
- Rabbani MA, Maruyama K, Abe H, Khan MA, Katsura K, Ito Y, Yoshiwara K, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2003) Monitoring expression profiles of rice genes under cold, drought, and high-salinity stresses and abscisic acid application using cDNA microarray and RNA gel-blot analyses. Plant Physiol 133: 1755–1767PubMedCrossRefGoogle Scholar
- Reddy AR, Ramakrishna AC, Sekhar AC, Nagablushana I, Ravindra Babu P, Bonaldo MF, Soares MB, Etennetzen JL (2002) Novel genes are enriched in normalized cDNA libraries from drought-stressed seedlings of rice (Oryza sativa L. subsp.indica cv. Nagina 22). Genome 45: 204–211PubMedCrossRefGoogle Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual, Ed 2. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
- Seki M, Namsaka M, Ishida J, Nanjo T, Fujita M, Oono Y, Kamiya A, Nakajima M, Enju A, Sakurai T, Satou M, Akiyama K, Taji T, Yamaguchi-Shinozaki K, Caminci P, Kawai J, Hayashizaki Y, Shinozaki K (2002) Monitoring the expression profiles of 7000Arabidopsis genes under drought, cold and high-salinity stresses using a full-length cDNA microarray. Plant J 31: 279–292PubMedCrossRefGoogle Scholar
- Shinozaki K, Yamaguchi-Shinozaki K (1999) Molecular responses to drought stress.In K Shinozaki, K Yamaguchi-Shinozaki, eds, Molecular Responses to Cold, Drought, Heat and Salt Stress in Higher Plants. RC Lan-des, Austin, pp 11–28Google Scholar