53 genotypes of embryonal suspensor masses (ESMs) rescued from mature seeds of Norway spruce (Picea abies (L.) H. Karst.) were examined for their pattern of growth and development under standardized culture conditions in vitro. Patterns were classified according to the colour of the colonies grown in darkness, clarity of cell masses and proembryos in the mucilaginous ESM, surface boundary topology of colonies, structure of suspensors, growth rate of the ESM, and recovery of mature embryos.
Five distinctive major growth patterns were observed among ESM colonies under standardized culture conditions. The multiplication of proembryos and early embryos by cleavage and budding polyembryony was the main factor contributing to proliferation and colony growth and further determined the morphology of the colonies. Callus and teratological structures were induced from early embryos by changing the standardized culture conditions i.e. inadequate subculture, excessive dose of 2,4-D in the medium and premature exposure of the colonies to light. Results enable the selection of ESM genotypes for the predictable recovery of mature somatic embryos.
After 64 days of aseptic culture, germlings of Pinus Syvestris L. were cut at the middle of the hypocotyl and above the root. The upper and lower halves of the hypocotyls were transferred onto agar medium RM-196 of Linsmaier & Skoog (1964) including 2 mg/l IAA and 0.1, 1 or 10 mg/l kinetin, one or both halves being put in each vial. Callus growth and root formation was observed after 55 days.
The lower ends of basally cut seedlings generally formed callus tissue and 20% of them also formed roots from this callus. No roots and less callus growth were observed in the lower hypocotyle halves excised at both ends. In the latter hypocotyles callus growth was promoted by the presence in the same vial of a basally excised germling, including cotyledons and plumule. Increasing amounts of kinetin slightly enhanced callus formation of basally excised germlings but seemed to inhibit callus growth in hypocotyls excised at both ends and placed alone on the growth medium. The total amount of callus was greatest in hypocotyls which included intact cotyledons and plumule.
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