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Chapter 14 : Plant Cell Culture
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Plant cell culture has found wide applications ranging from studies on basic plant biochemistry and molecular biology to mass propagation and genetic engineering of crop species. This chapter talks about basic requirements for common procedures in plant cell culture. Successful plant cell culture requires extremely close attention to proper sterile technique and temperature control. Some laboratories use containment hoods when introducing microbes into plant cell cultures, such as during genetic transformation or inoculation with plant pathogens. Continuous light is sometimes detrimental to whole-plant growth and is rarely used in plant cell cultures. Many methods have been used to estimate the growth of plant cell cultures both directly and indirectly. The method chosen will depend on the requirements for maintaining the sterility of the culture and the speed and accuracy of the measurement. Assays in which the stain (such as fluorescein diacetate [FDA] or triphenyltetrazolium chloride [TTC]) is meant to be taken up only by living cells are known to sometimes give false-positive results with cells that no longer divide. One application of plant cell cultures is the study of plant secondary-metabolite biosynthesis and the commercial production of such valuable natural products.
Growth of opium poppy cell suspension cultures in 250-ml shake flasks. The inoculum (Papaver somniferum L. cv. Marianne, PBI 2009) ( 29 ) was 10 g (wet fresh weight) (13% [wt/vol]) per 75 ml of medium per 250-ml flask. Cells were cultured at 25°C and 150 rpm. The medium was 1B5C (B5 salts supplemented with 20 g of sucrose per liter, 1 mg of 2,4-D per ml, and 1 g of casein [NZ-Amine] per liter; pH 5.5). The initial sucrose level was less than 20 g/liter, due to dilution by sucrose-depleted medium in the inoculum.
Composition of common plant cell culture media
Growth regulators and additives commonly used in plant tissue cultures
Preparation of SH frozen stock a