Cloning and Expression of Calmodulin Genes Regulating Phytoalexin Production in Carrot Cells

Ishigaki, Eriko; Asamizu, Tetsuya; Arisawa, Munehisa; Kurosaki, Fumiya

doi:10.1248/bpb.27.1308

Cited by 17 publications

(26 citation statements)

References 19 publications

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“…12) Mycelial walls of the fungus Chaetomium globosum were prepared from 3-d-old culture, 15) and oligogalacturonides were prepared by the partial hydrolysis of the pectin fraction of carrot 12) according to the previously reported methods, respectively. 2-Chloroethylphosphonic acid was purchased from Sigma.…”

Section: Methodsmentioning

confidence: 99%

“…Cloning of CAM genes The suspension of the fungal mycelial walls (500-ml aliquots, 10 mg/ml H 2 O) was carefully sprinkled over carrot seedlings (3 weeks old, 100 mg), and, after incubation for 6 h, 12) total RNA was isolated from the cells with the RNeasy Plant Mini Kit (Qiagen). The RNA obtained (approximately 5 mg) was subjected to the rapid amplification of cDNA end (RACE) methods using the GeneRacer Kit (Invitrogen) after the generation of cDNA template by reverse-transcription (RT) reaction with AMV RT.…”

Section: Methodsmentioning

confidence: 99%

“…28, No. 6 oligogalacturonides, or 2-chloroethylphosphonic acid according to the method described previously, 12) and, at regular intervals, total RNA was isolated from 100 mg of the whole tissues as described above. In a separate experiment, RNA was prepared from the roots, leaves, stems of carrot seedlings and from ungerminated carrot seeds.…”

Section: Methodsmentioning

confidence: 99%

“…Expression of Carrot CAM Genes To elucidate the tissue-specific expression of CAM genes, 12) changes in their transcriptional levels in various carrot tissues were examined using RT-PCR. As shown in Fig.…”

Section: Cam Genes From Mycelial Wall-treated Carrot Eleven New Genesmentioning

confidence: 99%

“…Those results strongly suggested that carrot CAM should be composed of several isoforms, although only one CAM gene, cam-1, had been reported 11) from this plant. We have recently isolated several clones of carrot CAM genes and demonstrated 12,13) that the transcriptional activation of the specific CAM genes is induced in carrot cells upon the treatment with either oligogalacturonide elicitor or ethylene. It was also confirmed that more than 10 CAM genes occur in carrot cells using Southern blot analysis, and these observations suggested the possibility that, as is in oligogalacturonide-and ethylene-treated carrot, induction of the defense reactions in mycelial wall-treated carrot are evoked by the elevated expression of certain CAM genes but not mediated by "constitutively expressed" CAM protein.…”

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confidence: 99%

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Multiple Forms of Calmodulin Genes in Carrot Treated with Fungal Mycelial Walls

Ishigaki

Sugiyama

Kurosaki

2005

Biological & Pharmaceutical Bulletin

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Eleven cDNA clones encoding the open-reading frame of calmodulin and calmodulin-like proteins were isolated from carrot seedlings treated with fungal mycelial walls. These calmodulin genes were mainly expressed in the stems and leaves of carrot, although the transcriptional level was almost negligible in the seeds and root tissues. Based on nucleotide homology, these genes were divided into two classes, class I and class II, and reverse transcription-polymerase chain reaction analyses revealed that the expression level of class II genes transiently increased when carrot seedlings were treated with the mycelial walls. In contrast, the level of class I genes apparently did not show any significant change even after treatment with the fungal components. These results suggest that the defense responses induced in the fungi-infected carrot accompany the increase in the transcriptional levels of a certain calmodulin gene(s).

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Cam Genes From Mycelial Wall-treated Carrot Eleven New Genesmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Multiple Forms of Calmodulin Genes in Carrot Treated with Fungal Mycelial Walls

Ishigaki

Sugiyama

Kurosaki

2005

Biological & Pharmaceutical Bulletin

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Oligogalacturonides enhance cytokinin-induced vegetative shoot formation in tobacco explants, inhibit polyamine biosynthetic gene expression, and promote long-term remobilisation of cell calcium

Falasca

Capitani

Rovere

et al. 2007

Planta

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Long-sized oligogalacturonides (OGs) are cell wall fragments that induce defence and developmental responses. The Ca(2+)-dependent "egg-box" conformation is required for their activity, and polyamines may prevent them from adopting this conformation. Although OGs are known to inhibit auxin-induced growth processes, their effect on cytokinin-induced ones requires investigation. In the present work OGs were shown to promote cytokinin (benzyladenine, BA)-induced vegetative shoot formation from tobacco leaf explants, independent of the presence of CaCl(2) in the medium and of auxin (indoleacetic acid, IAA) supply. The effect of polyamines, putrescine (PU) and spermidine (SD) supplied with/without their biosynthetic inhibitors (DFMO, CHA) was also investigated, and showed that spermidine enhanced adventitious vegetative shoot formation, but only on medium containing Ca(2+) and IAA. Treatments with inhibitors blocked this promotive effect. OGs did not alter free polyamine concentrations, but caused a moderate increase of conjugated ones, and exhibited an early inhibitory effect on polyamine biosynthetic gene expression. OGs, but not SD, caused long-term changes in calcium-associated epifluorescent signals in the cell walls, and, later, inside the cells of specific tissues. Electron microscopy analysis (ESI system) demonstrated that calcium accumulated in the cell walls and vacuoles of OG-cultured explants. The relationship between OGs, cytokinin, calcium, and polyamines in adventitious vegetative shoot formation is discussed.

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Interrelationship between calmodulin (CaM) and H2O2 in abscisic acid-induced antioxidant defense in the seedlings of Panax ginseng

et al. 2012

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Calmodulin (CaM), the predominant Ca(2+) receptors, is one of the best-characterized Ca(2+) sensors in all eukaryotes. In this study the role of CaM and the possible interrelationship between CaM and hydrogen peroxide (H(2)O(2)) in abscisic acid (ABA) induced antioxidant defense were investigated in the seedling of Panax ginseng. Treatment of ABA (100 μM) and H(2)O(2) (10 mM) increased the expression of Panax ginseng calmodulin gene (PgCaM) and significantly enhanced the expression of the antioxidant marker genes such as superoxide dismutase, ascorbate peroxidase, glutathione reductase and the activities of chloroplastic and cytosolic antioxidant enzymes. Pretreatments with two CaM antagonists, trifluoperazine (TFP), N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide hydrochloride (W7) and inhibitor or scavenger, diphenyleneiodonium chloride, and dimethylthiourea of reactive oxygen species almost completely suppressed the up-regulation of antioxidant and PgCaM gene. Moreover, H(2)O(2) production and CaM content was almost completely inhibited by pretreatments with two CaM antagonists. In addition, the expressions of PgCaM gene under different biotic stress were analyzed at different time intervals. Thus it may suggests that CaM are involved in ABA-induced increased expression of PgCaM which triggers H(2)O(2) production through activating trans-plasma membrane NADPH oxidase, resulting in up-regulation of defense related antioxidant gene and also plays a pivotal role in defense response against pathogens.

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Cloning and Expression of Calmodulin Genes Regulating Phytoalexin Production in Carrot Cells

Cited by 17 publications

References 19 publications

Multiple Forms of Calmodulin Genes in Carrot Treated with Fungal Mycelial Walls

Multiple Forms of Calmodulin Genes in Carrot Treated with Fungal Mycelial Walls

Oligogalacturonides enhance cytokinin-induced vegetative shoot formation in tobacco explants, inhibit polyamine biosynthetic gene expression, and promote long-term remobilisation of cell calcium

Interrelationship between calmodulin (CaM) and H2O2 in abscisic acid-induced antioxidant defense in the seedlings of Panax ginseng

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