The frst systematic attempt was made by Hannig

(1904), who cultured mature embryos of two crucifers, Cochleria and

Raphanus.
ieterich (19!4) pointed mature embryos grew normally but those e"cised from
immature
seeds failed to achie#e the organi$ation of a mature embryo% &nstead,
they grew directly into seedlings, s'ipping the stages of normal embryogenesis%
(aibach (19!), 19!9) &n the interspecifc cross Linum
perenne x L. austriacum, (aibach noted that the seeds were greatly
shri#elled, #ery light, and incapable of germination% *y e"cising embryos
from such seeds and growing them on moist flter paper or on cotton
wadding containing sucrose he was able to raise the hybrid plants%
The mature embryos can be isol
a t e d w i t h r e l a t i # e ease by s p l i t t i n g open t h e seed% +eeds w i t h a h a r d
seed,coat are dissected a f t e r soa'ing them in w a t e r % +maller embryos re-uire
careful dissection w i t h t h e aid of a stereoscopic microscope and
-uic' t r a n s f e r to t h e c u l t u r e #ial% (iu et al% (199.a) found i t n e c e s s a r y to
c a r r y out dissections of globular embryos of Brassica juncea
most important aspect of embryo culture is the selection of the
right culture medium that would support progressive and orderly
development
of embryos excised at diferent stages of development. Early reports
of embryo culture were generally concerned with the development
of plants from mature embryos (post-germinal development) on a simple
medium. Hannig (1!") used a mineral salts-sucrose solution to culture
mature embryos (# mm long) of crucifers. $aibach (1#%) reared full
plants from excised hybrid embryos (1 mm long) using only 1%& glucose
solution. 'n contrast( immature embryos generally fail to grow on such a
simple medium. )heir nutritional re*uirements are more elaborate than
those of the mature embryos.
that would fa#our both good growth and
high sur#i#al% He altered the concentration of e#ery salt in the /+ medium
to study the e0ect of such #ariations, and on the basis of these e"periments
de#eloped a medium on which the growth of embryos was as
good as on /+ but sur#i#al was high (see 1ig% 11%2)% The composition of
the new medium is gi#en in Table 11%.% 3ompared with the inorganic
composition of /+ medium, /onniers medium has high concentrations of
4 5 and 3a !5 and a reduced le#el of 6H4 5 ions%
3asein hydrolysate (3H), an amino acid comple", has been widely used
as an additi#e to the embryo culture media% +anders and *ur'holder
(1947) tried it for the culture of pre,heart,shaped (100,!)0 8m long) embryos
of two species of Datura. 9hen added to the basal medium
(containing mineral salts, #itamins, and sucrose) with cysteine and tryptophan,
3H increased embryogenic di0erentiation and the si$e of the embryo%
&n fact, au"in is generally inhibitory for
embryo growth (:agha#an, 1970)% /onnier (19;7) suggested that hormones
should not be added to the embryo culture media because they
bring about structural abnormalities% He felt that embryos are autonomous
for most of the growth regulators% This has been clearly shown at
least for <=. in Phaseolus, where the suspensor fulfls the re-uirement of
e"cised embryos for this hormone (see +ection 11%4)%