Isolation of cDNAs for hardening-induced genes from Chlorella vulgaris by suppression subtractive hybridization

Takeshi Machida, Hisako Murase, Eri Kato, Ken-ichi Honjoh, Kiyoshi Matsumoto, Takahisa Miyamoto, Masayoshi Iio

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Several organisms acquire freezing tolerance by altering intracellular reactions under sublethal low temperatures. Although many researchers have reported that organisms change their intracellular functions, such as transcript level and enzymatic activity, under stress conditions, acquisition mechanisms of freezing tolerance are still poorly clarified. In this study, we attempted to identify novel hardening-induced genes from Chlorella vulgaris C-27, a frost-hardy strain. A PCR-based suppression subtractive hybridization library was generated and the corresponding cDNA clones were isolated. A total of 263 unique cDNA clones were obtained and sequenced. Homology analysis showed that 64 distinct known proteins were encoded by the respective clones. The expression patterns of 29 of the genes were analyzed by using qPCR. Especially, six genes, which respectively encode two late embryogenesis abundant (LEA) proteins (HIC6 and HIC12), NADPH thioredoxin reductase (NTR), chlorophyll a/b-binding protein (Cab), ζ-carotene desaturase, and Nip7, showed remarkable increase in transcripts over 100 times greater than those of unhardened cells. We discuss the possible contribution of the genes, which showed remarkable transcriptional increases, in acquisition of freezing tolerance of Chlorella.

Original languageEnglish
Pages (from-to)238-246
Number of pages9
JournalPlant Science
Volume175
Issue number3
DOIs
Publication statusPublished - Sep 1 2008

Fingerprint

Chlorella vulgaris
suppression subtractive hybridization
Complementary DNA
cold tolerance
Freezing
Clone Cells
clones
Genes
genes
carotene 7,8-desaturase
Chlorophyll Binding Proteins
Thioredoxin-Disulfide Reductase
Chlorella
organisms
Carotenoids
frost
NADP (coenzyme)
Libraries
Embryonic Development
Proteins

All Science Journal Classification (ASJC) codes

  • Genetics
  • Agronomy and Crop Science
  • Plant Science

Cite this

Isolation of cDNAs for hardening-induced genes from Chlorella vulgaris by suppression subtractive hybridization. / Machida, Takeshi; Murase, Hisako; Kato, Eri; Honjoh, Ken-ichi; Matsumoto, Kiyoshi; Miyamoto, Takahisa; Iio, Masayoshi.

In: Plant Science, Vol. 175, No. 3, 01.09.2008, p. 238-246.

Research output: Contribution to journalArticle

Machida, Takeshi ; Murase, Hisako ; Kato, Eri ; Honjoh, Ken-ichi ; Matsumoto, Kiyoshi ; Miyamoto, Takahisa ; Iio, Masayoshi. / Isolation of cDNAs for hardening-induced genes from Chlorella vulgaris by suppression subtractive hybridization. In: Plant Science. 2008 ; Vol. 175, No. 3. pp. 238-246.
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