Studies on the Growth in Culture of Plant Cells

Stuart, R.; Street, H. E.

doi:10.1093/jxb/22.1.96

Cited by 76 publications

(13 citation statements)

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“…It has also been reported that some medium supplements, such as amino acids, ammonium ion, and purine derivatives promote cell proliferation (17)(18)(19). We, however, observed no dispersal over several fractions or serious decrease in the mitogenic activity of CMFs in the purification experiments that would be expected if the CMF was composed of several components acting synergistically.…”

Section: Discussioncontrasting

confidence: 56%

Phytosulfokine, sulfated peptides that induce the proliferation of single mesophyll cells of Asparagus officinalis L.

Matsubayashi

Sakagami

1996

Proc. Natl. Acad. Sci. U.S.A.

450

293

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Proliferation of dispersed plant cells in culture is strictly dependent on cell density, and cells in a low-density culture can only grow in the presence of conditioned medium (CM). No known plant hormones have been able to substitute for CM. To quantify the mitogenic activity of CM, we examined conditions for the assay system using mechanically dispersed mesophyll cells ofAsparagus officinalis L. and established a highly sensitive bioassay method. By use of this method, the mitogenic activity of CM prepared from asparagus cells was characterized: it was heat-stable, susceptible to pronase digestion, and resistant to glycosidase treatment. On the basis of these results, the mitogenic activity in CM was purified 107-fold by column chromatography, and two factors named phytosulfokine-a and -13 (PSK-a and PSK-f8) were obtained. By amino acid sequence analysis and mass spectrometry, the structures of these two factors were determined to be sulfated pentapeptide (H-Tyr(SO3H)-IleTyr(SO3H)-Thr-Gln-OH) and sulfated tetrapeptide (HTyr(SO3H)-Ile-Tyr(SO3H)-Thr-OH). PSK-ac and PSK-p8 were prepared by chemical synthesis and enzymatic sulfation. The synthetic peptides exhibited the same activity as the natural factors, confirming the structure for PSK-a and PSK-p mentioned above. This is the first elucidation of the structure of a conditioned medium factor required for the growth of low-density plant cell cultures.The proliferation of plant cells in dispersed culture is strictly dependent on the initial cell density, and mitotic activity in low-density suspension cell cultures cannot be stimulated by supplementation with known plant hormones or defined nutrients. However, proliferation of plant cells at low density is induced by the addition of conditioned medium (CM) prepared from rapidly growing cells in culture (1). A possible explanation for this phenomenon is that a mitogenic factor or factors produced by the individual cells is essential for cell proliferation.Since the late 1980s, detection and characterization of mitogenic factors that are often called conditioned medium factors (CMFs) have been performed in a few culture systems: highly hydrophilic, neutral compounds in CM of maize cell line (2, 3); highly hydrophilic, relatively heat-stable compounds in CM of a carrot cell line (4); and pronase-resistant, relatively small compounds in CM of Pinus radiata cells (5) (Takii Shubyo, Japan) were planted on moist sterile soil and kept in a growth room at 25 ± 2°C under a daily 16-h light period [Toshiba (Tokyo) DR 400/T and Mitsubishi (Yokohama) MLRBOC 400 F-U lamps; -20,000 lux at plant level]. The cladodes of 40-to 60-day-old spears were used as the experimental material. Cell lines of Zea mays L. and Oriza sativa L. were maintained with weekly subculturing in Murashige and Skoog medium supplemented with 1.0 mg of 2,4-dichlorophenoxyacetic acid per liter and 30 g of sucrose per liter. Cultures were incubated at 25°C in the dark, with rotary shaking at 120 rpm.Bioassay. Approximately 0.5 g of cladodes were steri...

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Section: Discussioncontrasting

confidence: 56%

Phytosulfokine, sulfated peptides that induce the proliferation of single mesophyll cells of Asparagus officinalis L.

Matsubayashi

Sakagami

1996

Proc. Natl. Acad. Sci. U.S.A.

450

293

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“…Favorable effect of nurse cultures, feeder layers, or conditioned media suggested the presence of division-inducing factors that are secreted into the medium with cultured cells [17,18]. Many efforts have been made to identify and characterize such factors and initial amino acids, purine derivatives, highly hydrophilic and heat stable agents, or protease-resistant small compounds were considered as putative mitogenic inducers; however, the available assay methods were not sufficiently sensitive to define any specific compound [19][20][21]. The elaboration of highly sensitive bioassay systems for the characterization of mitotic activity in low-density cultures of asparagus in the presence of conditioned medium led to the discovery of phytosulfokines (PSK), biologically active components with mitogenic activity [22].…”

Section: Digital Signaturementioning

confidence: 99%

Early studies on the effect of peptide growth factor phytosulfokine-α on Brassica oleracea var. capitata L. protoplasts

Kiełkowska

Adamus

2017

Acta Soc Bot Pol

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Phytosulfokines (PSK) are peptidyl growth factors with the potential of inducing cell proliferation. We examined the effect of supplementation of liquid culture medium with 0.1 µM phytosulfokine-α (PSK-α) on protoplast viability and division frequencies in seven accessions of Brassica oleracea var. capitata L., including cultivars and breeding lines. Protoplasts were isolated from leaves and hypocotyls of in vitro grown plants and immobilized in calcium-alginate layers. Cabbage protoplast-derived cells cultured in medium supplemented with 0.1 µM of PSK-α had higher viability and division frequencies compared to cells cultured in PSK-α-free control medium. The effect of PSK-α was more pronounced in low-responding accessions ('Sława z Gołębiewa' , 'Ramkila F1' , LM, and LM98); however, in two cultivars with very low response ('Badger Shipper' and 'Oregon 123'), although the division frequencies in the media supplemented with PSK-α were increased over the control, the differences were not significant. Obtained callus colonies were subjected to regeneration. PSK-α supplemented into the liquid culture medium had an indirect effect on shoot regeneration by inducing sustained cell divisions leading to an increase in shoot regeneration in Sława z Gołębiewa and both breeding lines.

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“…This is called the minimum effective density. A conditioning factor is present in calli or cultured medium that induces cell growth, resulting in a lower value for the minimum effective density (Stuart and Street 1969). Yamakawa et al (1985) also reported that the conditioning factor present in a tobacco cell suspension culture decreased plating grape cell densities.…”

Section: Introductionmentioning

confidence: 99%

Effects of conditioning factor on anthocyanin production in strawberry suspension cultures

et al. 1994

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To investigate the effects of the conditioning factor on anthocyanin production in the cultured strawberry, Fragaria ananassa cv Shikinari, cells were cultured in LS medium supplemented with a filtered conditioned broth. Anthocyanin accumulation was greatly enhanced using media conditioned for 5-8 days. This effect was promoted by increased concentrations of the conditioned broth. The amount of anthocyanin produced in a 100% conditioned broth was about eight times higher than in the control broth after 5 days. The substance that promoted anthocyanin accumulation in the conditioned medium was thermostable, because neither cell growth nor anthocyanin accumulation were affected by boiling for 10 min. Anthocyanin production was further enhanced by adding freeze-dried conditioned broth to freshly prepared LS medium ; however, this conditioned broth affected cell growth only in the early phase of culturing. The content of peonidin-3-glucoside, a major anthocyanin, increased as the number of days the cell growth proceeded, while that of cyanidin-3-glucoside decreased. The contents of these two major anthocyanins were also changed by adding a preculture broth as a conditioning factor.

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Studies on the Growth in Culture of Plant Cells

Cited by 76 publications

References 0 publications

Phytosulfokine, sulfated peptides that induce the proliferation of single mesophyll cells of Asparagus officinalis L.

Phytosulfokine, sulfated peptides that induce the proliferation of single mesophyll cells of Asparagus officinalis L.

Early studies on the effect of peptide growth factor phytosulfokine-α on Brassica oleracea var. capitata L. protoplasts

Effects of conditioning factor on anthocyanin production in strawberry suspension cultures

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