Patent GB-760112-A (GB)
Title: Improvements in or relating to cultivation of plant tissue
IPC: A01H4/00
Filed: 1953-05-12
Abstract: Oxalic acid, coumarin, melilotic acid and Vitamin C are produced by inoculating a nutrient aqueous medium with vigorouslygrowing undifferentiated plant cells (defined) and maintaining said plant cells under conditions of submerged agitated aeration in the presence of an embryonic type cell stimulant (see Group VI).  For the production of oxalic acid, Rumex acetosa tumor tissue is subcultured on solid nutrient agar until rapidly-growing undifferentiated cells are obtained, the cells are then transferred to an aqueous nutrient medium which is subjected to agitated aeration under sterile conditions at 25 DEG  C. for about two weeks.  The tissue was filtered, dried at 100 DEG  C., powdered and extracted with acidified water.  For the production of coumarin and melilotic acid, sweet clover seeds were planted on nutrient agar medium containing inter alia p-chlorophenoxyacetic acid and yeast extract, subcultured until vigorously growing tissue developed, which was then grown in an aqueous nutrient medium containing p-chloro-phenoxyacetic acid and yeast extract under submerged aeration and sterile conditions for about three weeks at room temperatures.  The tissue was filtered off and dried to produce a product containing coumarin and melilotic acid.  For Vitamin C, a crown gall is removed from a tobacco plant and is subcultured on nutrient agar until vigorously growing cells are obtained which are then grown for one month in an aqueous nutrient medium under conditions of submerged aeration.  The filtered tissue after freeze drying and grinding contained ascorbic acid. Leaf tissue of Agare tonmeyana grown in media containing p-chlorophenoxyacetic acid and yeast extract produced a product useful as a source of steroid compounds.ALSO:Finely divided masses of the cells of plants classified above the Thallophytes in septematic botany, are produced by inoculating a nutrient aqueous medium with vigorously growing undifferentiated plant-cells (defined), and maintaining said plant cells under conditions of submerged agitated aeration in the presence of an embryonic-type cell stimulant which is (a) a virus infection e.g. Aureogenus magnivena which causes tumors in many plants (b) a bacterial infection e.g. crows galls caused by Agrobacterium tumefaciens (c) an excretion from certain insects such as larvae of Gnovemoschema gallaesolidaginis and (d) a plant growth hormone e.g. indoleacetic acid, indolebutyric acid, naphthalene acetic acid and p-chlorophenoxyacetic acid.  The aqueous medium contains (a) one or more carbohydrates such as glucose, sucrose, corn syrup, starch and dextrose (b) mineral salts such as phosphates, nitrates, chlorides and sulphates of the metals sodium, potassium, calcium and magnesium (c) trace elements and (d) vitamins such as thiamine, riboflavin, pyridoxine hydrochloride, pantothenic acid and niacin, as such, or in the form of crudes e.g. yeast extract.  To prevent clumping during growth the enzyme pectase is added to the nutrient medium.  Growth is at 20-30 DEG C with aeration at least a quarter of a volume of air per volume of culture medium per minute. Growth is for 5 to 7 days.  The process is applicable to the production of (a) food or fodder and (b) valuable materials such as vitamins, steroids, alkaloids, antimicrobial agents, sugars, enzymes, organic acids and aromatic materials (see Group IV(b)) which may be recovered from the culture medium or from the cells of the plant tissue.