Somatic Embryogenesis Using Cucumis sativus (L.) Cotyledons
N.M.P. Guedes* and P.H. Jennings, Dept. of Horticulture, Kansas State Univ.,
Manhattan, KS 66502
To improve somatic embryogenesis of Cucumis sativus, two types of ex-
plants (cotyledons and stem sections) were cultured on Murashige and Skoog
(MS) media supplemented with 2,4-D (2.0 mg•L–1) + kinetin (0.5 mg•L–1). After
4 weeks, the embryogenic callus was transferred for 2 weeks to MS + NAA (1.0
mg•L–1) for embryo development. Stem sections failed to develop embryos while
cotyledons responded with 14% embryo formation. The embryos were trans-
ferred to MS without hormones for 4 weeks to allow for plantlet growth. These
embryos developed only shoots. To improve on the successful generation of
embryos with root and shoot development, the procedures used above were re-
peated, but the cotyledons were cut into three sections to be used as explants.
Each transverse section of the cotyledon was approximately 2–3 mm wide. All
sections produced callus but not all of them were embryogenic. From the first
section (cotyledon base), the second (between the first and third section) and the
third section (furthest from the cotyledon base), respectively, 58%, 31%, and 5%
embryo development occurred. Those embryos from the basal cotyledon sec-
tions regenerated 10 plantlets, 5 with shoots and roots and 5 with only shoots.
Approaches to enhance somatic embryogenesis, and shoot and root develop-
ment, will be discussed.
379 (PS 8)
Axillary Shoot Proliferation from in Vitro Culture of Pulmonaria
L. Shoots
Dennis P. Stimart* and John C. Mather, Dept. of Horticulture, Univ. of Wiscon-
sin, 1575 Linden Dr., Madison, WI 53706
Actively growing shoots from Pulmonaria L. ‘Roy Davidson’ were cultured in
vitro on Murashige and Skoog medium containing benzyladenine (BA) to establish
proliferating cultures. BA at 0, 0.4, 0.8, 4.4, 8.8, and 44.4 µM was compared for
shoot proliferation and rooting response. Shoot count was highest on 8.8 µM BA
with root count highest on 0 or 0.4 µM BA. Subculture 4 weeks later of shoots to the
same treatments resulted in highest shoot counts on 44.4 µM BA. Optimum level for
micropropagation was 8.8 or 44.4 µM BA. Greatest rooting was at 0 or 0.4 µM BA.
380 (PS 8)
Determination of Optimum Nitrogen Ratios and Concentrations
for Embryo Cultures of Alstroemeria
Paula C. Moreck* and Mark Bridgen, Dept. of Plant Science, U-67, Univ. of Con-
necticut, Storrs, CT 06269.
The rates of in ovulo germination of embryos of three genotypes of Alstroemeria
were observed. Ovules were harvested ten days after pollination and cultured on
sixteen different treatments containing Murashige and Skoog (MS) basal me-
dium with no plant growth regulators. The media contained four different concen-
trations of total nitrogen: 20, 40, 60, 80 mM. Within these concentrations were
either 1:0, 1:1, 1:2, or 2:1 ratios of nitrate to ammonium. Standard MS medium,
with a concentration of 60 mM total nitrogen and a ratio of 2:1 nitrate to ammo-
nium, was used as the control. The overall rates of germination for all three geno-
types were low in all treatments. The percentage of zygotic germination was low
while the percentage of somatic embryos produced was high. In some situations,
callus or deformed embryos were produced. Effects of the treatments on embryo
development and germination will be discussed.
381 (PS 8)
Effect of Quantity and Depth of Media per Culture Container on
Shoot Production of Muscadine Grape Cultured in Vitro
Carol D. Robacker*, Dept. of Horticulture, Georgia Station, Griffin, GA 30223
Previous studies indicated that the number of shoots formed per nodal ex-
plant varied significantly depending upon the type of culture container used.
Amount of media per container and amount of time that media was autoclaved
were variables that differed among the containers. To determine the cause for the
container effect on shoot number, a study was conducted in which autoclave time
and amount of media per container were the same for all tested containers. Media
was autoclaved in 500-mL batches for 30 min, then poured in 30-mL aliquots
into sterile containers. Containers tested were plastic petri plates, GA-7 Magenta
vessels and glass jars. Depth of the medium was 5 mm in plates, 8 mm in ves-
sels, and 12 mm in jars. Nodes from in vitro grown shoots of ‘Triumph’, ‘Regale’,
and ‘Fry’ were cultured on Murashige and Skoog salts and vitamins with 2 mg•L–1
BA and 8 g•L–1 agar. Results indicated that the greatest number of shoots formed
in the jars. In a second study, nodes were cultured on petri plates containing 30
mL of medium (depth 5 mm) or 70 mL (depth 12 mm). Two to three times more
shoots formed on the plates with the greater amount of medium. These studies
indicate amount and depth of medium are factors influencing shoot number.
This has some really interesting information here.
I do a lot of reading and studying of plant pathology, reproduction, cell structure and division. As well as mutations.
So much cool work being done.
I’m sure most of you could careless about this stuff.
I can post more interesting finds ,if anyone is into reading and thinking. This stuff is like so clear you can see it in your head. Idk maybe just me.
Thanks @MK3_Pharms.
Yep this was the one. Thanks for changing it. I hate reading threads I can’t respond too. I should just switch to commercial side but I don’t find I care for the attitude of a lot of the people on that side. I’m comfy here. Lol.
This makes total sense to me. I have broken many cots off removing seed caps and buried both pieces to end up with 2 plants. 1 from the stem root and one from the cots.
They feed the plant for weeks so the embryonic matter is full of nutritional value that the plant can survive on and should spring growth if buried.
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Its very informative. I get all kinds of stuff sent to me. Most of my days ate spent reading stuff like this.