Embryogenesis from coffea leaf segments through the two step method is indicated in figures 1 to 18.

They were resulted from cultures on secondary medium conaining 0.5 µM NAA and 2.5 µM KIN.

1. A general surface morphology of a cluster of friable embryogenenic callus undergoing the

differentiation
2. The callus surface was rough in appearance and yellow in colour. The callus was comprised of two
different parts: one was a friable yellowish callus and the other, a brownish callus. The friable
yellowish callus is composed of cells with a spherical shape and the brownish callus consist of
elongated cells and a membranous layer covering them
3. Sometimes, the spherical cells are also covered with a a membranous layer and fibril like
structures from over the embryogenic callus.
4. The arrangement of both spherical isodiametric cells are elongated tubular cells in clearly
illustrated
5. The large nodules are composed of spherical cells
6. Under a high magnification, the arrangement of isodiametric spherical cells on the surface of a
cluster of friable embryogenic callus can be easily observed. This spherical structure is a growing,
putative proembrionic stage derived from a single cell of about 15µm in diameter
7. Part of the callus mass surface is often covered by a slime like membranous material and spherical
cells are interconnected by fibrils
8. Illustrated that two single cells are about 20µm in diameter and fibrils are interconnecting them
9. On the other hand, in the advanced stage of differentiation, spherical cells are sprouting from the
basal region of a developed embryo. This differentiation pattern clearly illustrates secondary
embryo formation from primary coffea somatic embryos.
10. It is interesting to note that early globular embryos already have fairly well defined suspensors.
11. A malformed elongated globular and a group of early globular embryos are seen from an
embryogenic callus cluster.
12. Illustrates the state that an embryo at a typical globular stage is emerging from a callus surface
still interconnected by a suspensor.
13. A malformed large embryo with the appearance of a bottle like shape and early globular embryos
at various stages of development are clearly seen within a cluster of embryogenic callus.
14. An early heart shape embryo with a slight depression on the tip and one slightly elongated from
globular embryo are depicted in figures 14
15. Shows the state that a typical heart shaped and globular shaped embryos are emerging from a
friable embryogenic callus located in the vicinity of well developed embryos. This developmental
pattern illustrates embryo formation at a border between developed embryos and friable
embryogenic callus.
16. A multicellular composition is definitely illustrated in a transverse view of the hand fractured
suspensor region of a somatic embryo.
17. The torpedo shape stage of coffea somatic embryos is shown in a concentrated form and the
presence of two cotyledonary primordial on the tip of embryos is visible.
18. Epidermis with relatively elongated cells is arranged with a single orientation on a heart shaped
embryo, which has the presence of two cotyledonary primordial.
19. somatic embryos at both the globular stage and the cotyledonary stage can be seen in the vicinity
of the leaf cut edge.
 20. There are three somatic embryos at the cotyledonary stage; two with well developed cotyledons,
and one malformed embryo with four cotyledons, which is seen in the upper right corner.
21. The asynchrony of coffea embryodevelopmentis illustrsted bt three embryos at different stages
of differentiation ; globular, elongated and cotyledonary shape. Well defined suspensors can be
clearly seen on cotyledonary embryos depicted on figs 19 to 23
22. Illustrated initial formation of a secondary embryo on the suspensor region
23. A magnified wiew of a suspensor structure of a coffea somatic embryo
24. Structural details of two cotyledonary leaves. Here, there is a relatively organized epidermis with
elongated cells in a row disposition.

Sequential coffea somatic embryo differentiation takea place through several developmental stages. It
proceeds through the initial single spherical cells (embryogenis mother cell); the development of globular
structures; heart stage; torpedo stage; and finally becomes a cotyledonary stage embryo. The suspensor
formation was early noticed as early as the globular stage of embryo differentiation.