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; M, Iio

doi:10.1016/j.plantsci.2008.04.002

Cited by 22 publications

(24 citation statements)

References 54 publications

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“…Seven genes including rbcL (clone 37), cell division gene ftsH (clone 39), psbB (clone 46), NADH dehydrogenase subunit nad6 (clone 69), atpB (clone E6), cox2 (clone B3), and initiation factor infA (clone F5) were selected for RT-qPCR. The β-actin gene was used as the internal standard (Machida et al, 2008). The length of amplified PCR fragments ranged from 100 to 200 bp.…”

Section: Gene Expression Profiles By Rt-qpcr Analysismentioning

confidence: 99%

Gene expression profiles of the heterotrophic microalga Chlorella pyrenoidosa F-9

Sun¹,

Xu²,

Jiang³

et al. 2014

Genet. Mol. Res.

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ABSTRACT.A strain of the microalga Chlorella pyrenoidosa F-9 in our laboratory showed special characteristics when transferred from autotrophic to heterotrophic culture. In order to elucidate the possible metabolic mechanism, the gene expression profiles of the autonomous organelles in the green alga C. pyrenoidosa under autotrophic and heterotrophic cultivation were compared by suppression subtractive hybridization technology. Two subtracted libraries of autotrophic and heterotrophic C. pyrenoidosa F-9 were constructed, and 160 clones from the heterotrophic library were randomly selected for DNA sequencing. Dot blot hybridization showed that the ratio of positivity was 70.31% from the 768 clones. Five chloroplast genes (ftsH, psbB, rbcL, atpB, and infA) and two mitochondrial genes (cox2 and nad6) were selected to verify their expression levels by real-time quantitative polymerase chain reaction. Results showed that the seven genes were abundantly expressed in the heterotrophic culture. Among the seven genes, the least increment of gene expression was ftsH, which was expressed 1.31-1.85-fold higher under heterotrophy culture than under autotrophy culture, and the highest increment was psbB, which increased 28.07-39.36 times compared with that under autotrophy conditions. The expression levels of the other five genes were about 10 times higher in heterotrophic algae than in autotrophic algae. In inclusion, the chloroplast and mitochondrial genes in C. pyrenoidosa F-9 might be actively involved in heterotrophic metabolism.

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Section: Gene Expression Profiles By Rt-qpcr Analysismentioning

confidence: 99%

Gene expression profiles of the heterotrophic microalga Chlorella pyrenoidosa F-9

Sun¹,

Xu²,

Jiang³

et al. 2014

Genet. Mol. Res.

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“…A few cDNA clones which represent ribosomal proteins such as 60S, 40S and 30S were known to be gene sequences respond towards stress. These ribosomal proteins play important roles in de novo protein synthesis (Machida et al 2008). Clones which have no significant similarity with any sequences in the databases were categorized as unknown genes (9%) and into others group (19%) that include cDNA clones that have no similarity with any nucleotide, mRNA or EST sequences in NCBI database (GR505498, GR505499, GR505500, GR505501, GR505502, GR505504, GR505505, GR505506, GR505507, GR505508, GR505509, GR505510, GR505511, GR505512, GR505513, GR505515, GR505516, GR505517), These sequences could be considered as novel genes induced by JA in kesum roots and further characterization must be done to identify their function in plant stress response, JA signalling and secondary metabolites production.…”

Section: Identification Of Ja-responsive Genesmentioning

confidence: 99%

Alteration of Abiotic Stress Responsive Genes in Polygonum minus Roots by Jasmonic Acid Elicitation

Ismail¹,

Gor²,

Mohamed‐Hussein³

et al. 2011

Plants and Environment

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“…11) Hardened cells of Chlorella acquire freezing tolerance, 12) and several intracellular changes during hardening have been reported, but the relationship between the changes and freezing tolerance are still poorly clarified. Hence we isolated a full-length cDNA encoding NTRC from Chlorella and characterized the NTRC protein expressed in E. coli.…”

Section: Expression Pattern Of a Chloroplast Nadph-dependent Thioredomentioning

confidence: 99%

Expression Pattern of a Chloroplast NADPH-Dependent Thioredoxin Reductase inChlorella vulgarisduring Hardening and Its Interaction with 2-Cys Peroxiredoxin

Machida

Kato

Ishibashi

et al. 2009

Bioscience, Biotechnology, and Biochemistry

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A chloroplastic NADPH-dependent thioredoxin reductase gene was identified from Chlorella vulgaris and designated CvNTRC. Mature CvNTRC protein (mCvNTRC) was expressed in Escherichia coli, and it showed both NADPH-dependent thioredoxin reductase (NTR) and thioredoxin (Trx)-like dithiol-disulfide oxidoreductase activities. The transcript of CvNTRC increased throughout 24-h hardening, whereas the encoded protein amount and total NTR activity decreased once and then increased during hardening. By in vitro pull-down assay, a 21.2-kDa protein bound to mCvNTRC was isolated and identified as a 2-Cys peroxiredoxin (2-Cys Prx) based on the N-terminal sequence. These data suggest that CvNTRC is maintained at a constant level during hardening and functions as an antioxidant with 2-Cys Prx in the acquisition of freezing tolerance of Chlorella.

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

Cited by 22 publications

References 54 publications

Gene expression profiles of the heterotrophic microalga Chlorella pyrenoidosa F-9

Gene expression profiles of the heterotrophic microalga Chlorella pyrenoidosa F-9

Alteration of Abiotic Stress Responsive Genes in Polygonum minus Roots by Jasmonic Acid Elicitation

Expression Pattern of a Chloroplast NADPH-Dependent Thioredoxin Reductase inChlorella vulgarisduring Hardening and Its Interaction with 2-Cys Peroxiredoxin

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