Studies on the Growth Requirements and Growth Measurements of Cell Cultures of <i>Haplopappus gracilis</i>

Eriksson, Tage

doi:10.1111/j.1399-3054.1965.tb06994.x

Cited by 169 publications

(62 citation statements)

References 17 publications

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“…Withdrawal of samples risks contamination and may result in an unrepresentative sample. Monitoring growth by measurement of turbidity of liquid cultures has been reported (1)(2)(3)8). I report here a method of measuring the growth of cultures grown in sidearm flasks (Fig.…”

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confidence: 99%

Turbidimetric Measurement of Plant Cell Culture Growth

Sung¹

1976

Plant Physiol.

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Turbidimetric measurement of cultures grown in sideann flasks was used to measure the growth of plant cells. The turbidity was shown to vary proportionaly with cell number and dry weight over time. The effect of different freezing conditions on the growth of the culture was presented to demonstrate the appliation of the sidearm-turbidity method.Growth of plant cells in liquid culture is usually determined by cell counting, packed cell volume, cell mass, or cellular protein content (6). These techniques involve tedious preparation and sacrificing of the cells. Withdrawal of samples risks contamination and may result in an unrepresentative sample. Monitoring growth by measurement of turbidity of liquid cultures has been reported (1)(2)(3)8). I report here a method of measuring the growth of cultures grown in sidearm flasks (Fig. 1) by the turbidity, which is determined through the sidearm with a Klett Summerson photoelectric colorimeter. Once turbidity has been calibrated against cell number, this technique allows growth to be measured easily and routinely in batch cultures. In this paper, I report experiments with carrot and soybean that demonstrate the method and present an example of its application. MATERIALS AND METHODSAll the cultures were grown in Linsmaier and Skoog (4) medium on a rotary shaker at 28 C. Soybean culture initiated from soybean stems (Glycine max (L.) Merr. cv. Kanrich) in this laboratory, was grown in medium supplemented with 3 mg/l NAA2 and 0.5 mg/l of 2-IP as the hormones. Wild carrot (Daucus carota L.) culture was obtained from H. E. Street, University of Leicester, England, and was grown in 0.1 mg/l 2,4-D.For growth determination, cultures were routinely grown in sidearm flasks (Medical Research Corp., Boston, Mass. 02121, nipped neck, 100 x 14 mm sidearm tube). The nipped neck allows incubation of large volumes of culture; 500-ml flasks will contain 200-ml culture; 250-mi flasks, 100-ml culture; and 125-ml flasks, 50-ml culture. Sidewise tilting of the flask fills the sidearm, so that turbidity can be read directly through the sidearm in a Klett Summerson colorimeter with a blue filter at 400 to 465 nm.For cell number determination, soybean suspension cultures were digested with Cr2O3* 5H20, 2.5%, at 60 C for 0. were treated with the same acids for 10 min each. Clumps were then dispersed with hypodermic needles of gauge numbers 20, 22, 26 and counted under the microscope using a cell counter of 0.2 mm in depth.Dry weight was determined by slowly pipetting 2 ml of cells onto a preweighed filter paper (Whatman no. 906, 21 mm diameter), placed on top of a deck of paper towels to absorb most of the liquid that passed through the filter paper. The cells and the filter paper were then dried overnight at room temperature and weighed.The size of aggregates was estimated by filtering cell suspensions through Nitex filters made of nylon sheet (Paul 0. Abbe Inc., Little Falls, N. J. 07424) of different porosity.For the freezing experiment, cultures at Klett 500 were frozen in growth mediu...

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Turbidimetric Measurement of Plant Cell Culture Growth

Sung¹

1976

Plant Physiol.

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“…The TTC test was performed essentially according to the method of Steponkus and Lanphear (17) except that the reaction temperature was 27 C. To determine the growth rate, frozen-thawed cells suspended in 10 ml of fresh medium were cultured in 1 00-ml Erlenmeyer flasks, under previously described conditions. The growth rate was estimated photometrically at 610 nm using a Bausch-Lomb colorimeter according to the method described by Eriksson (2).…”

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Survival of Suspension-cultured Sycamore Cells Cooled to the Temperature of Liquid Nitrogen

Sugawara

Sakai²

1974

Plant Physiol.

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(v/v) DMSO2 and 10% (w/v) glucose in the distilled water. Thus, this cryoprotective solution was used throughout this study.On the 5th to 8th cultural days after inoculation, cell suspensions to be frozen were collected in a graduated spitz tube (15 mm diameter, 110 mm long). To the packed cells of 0.5 ml, equal volume of the cryoprotective solution which consisted of 24% (v/v) DMSO and 10% (w/v)

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“…Eriksson [12] have concluded that pyridoxine and nicotinic acid are essential vitamins assist thiamine (when studying the optimum growth of Haplopappus gracilis on a modified MS medium) [9]. Decreased weight of maize calli was observed by addition of thiamine from 110mg/ml to 0mg/ml but by removing inositol and pyridoxine from MS medium does not show as an important factor in growth (Polikarpochkina et al) [13].…”

Section: Callus and Somatic Growthmentioning

confidence: 99%

Role of Vitamins in Plant Growth and their Impact on Regeneration of Plants under Invitro Condition

Tomar¹

2018

IJRASET

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Vitamins are the essential compounds synthesized and used by plants. In tissue culture media, the addition of vitamins is not that common because the amount needed by the plant is really unknown and it also varies from plant to plant. Vitamins show a direct and indirect effect on the growth of callus, somatic cell growth, rooting, and embryo development. Studies show that thiamine is associated with cytokinin and plays role in inducing root and callus growth. Thiamine also facilitates the production of more secondary metabolites such as proteases in pineapple. Biotin and riboflavin also play role in development of callus. There are many other vitamins which are essential or semi essential for the growth of plants of various types. Vitamin D causes uptake of calcium, cell elongation and meristematic cell division. Vitamin C is known to enhance the shoot and root growth. Nicotinic acid is required by plants for synthesizing the amino acids and also helps in carbohydrate metabolism. In this review we will discuss role of these vitamins in some plants that is associated with different plant species.

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Studies on the Growth Requirements and Growth Measurements of Cell Cultures of Haplopappus gracilis

Cited by 169 publications

References 17 publications

Turbidimetric Measurement of Plant Cell Culture Growth

Turbidimetric Measurement of Plant Cell Culture Growth

Survival of Suspension-cultured Sycamore Cells Cooled to the Temperature of Liquid Nitrogen

Role of Vitamins in Plant Growth and their Impact on Regeneration of Plants under Invitro Condition

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