Control of growth and cell division in plant cell suspension cultures

Abstract

STREET. 1973. Control of growth and cell division in plant cell suspension cultures. Can. J. Bot. 51: 1807-1823. In batch suspension cult~lres variation occurs in the growth and metabolism of the cells both in space and with time. Viable cell pop~~lations of sycamore (Acer pselrdol~lata~~lrs L.), showing greatly reduced aggregation and more uniform mor hology, can be obtained by incorporating enzymes into the culture medium. Such techniques cornbinefwith single cell cloning will take us closer to uniformity within the culture. The problem set by the continuous change in the metabolic activities of the cells with time (during the progress of the growth cycle of batch cultures) has been overcome by the use of low-density synchronous cultures and by the establishment of steady states of growth in chemostat cultures. Experimental work with sycalnore cell suspensions is described showing (1) the achievement of prolonged cell division synchrony in 4-liter suspension cultures and (2) the conformity of the growth kinetics of cells in chemostat culture to the mathematical model developed by Monod (1950, Ann. Inst. Pasteur (Paris), 79: 39&410) for microorganisnls. KING, P. J., K. J. MANSFIELD et H. E. STREET. 1973. Control of growth and cell division in plant cell suspensioil cultures. Can. J. Bot. 51: 1807-1823. Lors u'on cultive des suspensions de cellules en lot, des variations se produisent dans la croissance et le mtta%olisme des cellules en fonction de la situation spatiale et du temps. Des populations de cellules viables de sycomore (Acer pse~l~iaplam~lus L.) montrant une tendance B I'agrCgation considCrablement rCduite et une morphologie plus uniforme, peuvent Ctre obtenues en incorporant des enzymes dans le milieu de culture. De telles techniques, associkes B celles du clBnage de cellules isolCes, nous permettrons d'obtenir de plus en plus d'uniformitC dans les cultures. Le probEme, dCcoulant du changement constant des activitCs mCtaboliques des cellules en fonction du temps, (pendant le dkroulement du cycle de croissance dans les cultures en lot), a CtC surmontk par I'utilisation de cultures synchrones i faible densit6 cellulaire, et par le maintien d'un taux de croissance constant en culture en chemostat. Le travail expCrimental rCalisC sur les suspensions de cellules de sycomore est decrit; il montre qu'une synchronic prolongke des divisions cellulaires a Ct C obtenue sur des cultures de 4 litres de suspensions de cellules, et que la cinitique de la croissance des cellules cultivCes en chernostat obeit au modkle mathCmatique dCcouvert par Monod (1950, Ann. Inst. Pasteur (Paris), 79: 39@410), pour les microorganismes. Introduction(as growth in batch culture proceeds). Efforts This paper will examine two propos~t~ons ill have therefore been directed towards the dethe light of currellt research achievemellt from velopment of free-cell cultures and to the estabour own other laboratories. These proposi-lishment of cultural conditions leading to steady tions are (i) that plant cell suspension cultures states of grow...

Carbohydrate Metabolism and Activity of Pyrophosphate: Fructose-6-Phosphate Phosphotransferase in Photosynthetic Soybean (Glycine max, Merr.) Suspension Cells

“…Cultures of photoautotrophic cells derived from a number of species have been increasingly employed to study primary carbon metabolism (13,19,27). The type of metabolism displayed by a photosynthetic cell culture depends upon several parameters that include the light regime, supplemental phytohormones, inorganic and organic composition ofthe medium, and the metabolic capacities of the cells under study (19).…”

“…The type of metabolism displayed by a photosynthetic cell culture depends upon several parameters that include the light regime, supplemental phytohormones, inorganic and organic composition ofthe medium, and the metabolic capacities of the cells under study (19). The presence of an exogenous sugar source, such as sucrose, is a predominant factor influencing metabolism in cultured photosynthetic cells (18,19,27). For certain photosynthetic cells, the addition of sugar to the growth medium causes the cells to change from autotrophic metabolism to a "mixotrophic" type metabolism, where carbon is assimilated from the medium as well as CO2 (27 and references therein).…”

Growth and respiration of Petunia hybrida cells in chemostat cultures: A comparison of glucose-limited and nitrate-limited cultures

“…In GL cultures we measured a biomass protein content of 0.282 0.011 g/g DW, whereas the protein content of the nitrate-limited cultures could be calculated as 0.123 g proteirdg DW (Table 11). This value is rather low in comparison with most of the values reported for the protein content of the biomass in nitrogen-limited tobacco (0.16 g proteirdg DW) (Kato et al, 1980) and Acer pseudoplatanus (0.11-0.24 g proteirdg DW) cultures (King, 1976(King, , 1977King et al, 1973;Street et al, 1975;Young, 1973). On a fresh weight basis the protein contents of Petunia and tobacco cells were comparable (ca.…”

“…Most of these studies were carried out with Acer pseudoplatanus cultures at relatively low dilution rates (D = 0.003-0.009 h-') (King, 1976(King, , 1977King et al, 1973;Street et al, 1975, Young, 1973. Nitrate-limited growth was also studied in chemostat cultures of Nicotiana tabacum at relatively high dilution rates ( D = 0.018-0.040 h-') (Kato et al, 1980) and in sugar cane cultures at intermediate dilution rates (0.006-0.017 h-') (Veith and Komor, 1993).…”

“…The N sources used in these studies were either nitrate + urea (King, 1977;Street et al, 1975;Young, 1973), urea alone (King, 1976; Str6et et al., 1975), nitrate + ammonium (Kato et al, 1980), or nitrate alone (Veith and Komor, 1993). King et al (1973) did not specify the composition of the medium used in their study but claimed that their cultures were nitrate limited. Data presented by Street et al (1975) suggest, however, that King et al (1973 employed a medium containing nitrate + urea (see also King, 1977).…”