American Journal of Biochemistry and Biotechnology 7 (1): 32-42, 2011
ISSN 1553-3468
© 2010 Science Publications
32
Basal Salt Requirements Differ According to Culture Stage and
Cultivar in Date Palm Somatic Embryogenesis
Jameel M. Al-Khayri
Department of Agricultural Biotechnology, College of Agricultural and Food Sciences,
King Faisal University, Al-Hassa 31982, Saudi Arabia
Abstract: Problem statement: Various media formulations differing in basal salt composition are
arbitrary selected to provide essential nutrients for plant in vitro cultures. Evidence suggests that in
vitro response to various media formulations varies among genotypes and depends on the culture
stage. This study examined the efficacy of five basal salt formulations on callus growth and somatic
embryogenesis in date palm Phoenix dactylifera L. using three commercial cultivars, Khusab, Berny
and Barhee. Approach: Callus from shoot tip explants maintained on MS medium was introduced to
various media formulations including SH, W, MS, WPM and NN media containing 53.7 μM NAA and
7.4 μM 2iP. To assess the effect on callus growth, fresh callus weight was measured 4 and 8 weak
later. To evaluate embryogenesis response, callus was transferred to hormone-free media
corresponding to those during callus growth stage. Results: The optimum medium formulation varied
according to cultivar and culture stage. Extending callus growth to 8 weak allowed for greater
discernment of differences as compared to 4 weak because of the inherent slow growth nature of date
palm callus. The best callus growth was achieved in cv. Khusab using W and WPM media, cv. Berny
using SH and NN medium and cv. Barhee using SH, W and MS media. An optimal medium for callus
growth was not necessarily the best for somatic embryogenesis. The highest regeneration percentage in
cv. Berny occurred on WPM medium, cv. Khusab on W medium and cv. Barhee using W and WPM
media. The highest number of resultant embryos was achieved in cv. Khusab using W and SH media,
cv. Berny on WPM and MS media and cv. Barhee using W and SH media. Conclusion: This study
provides important information to optimize medium formulation in micropropagation protocols of
various date palm cultivars, particularly recalcitrant genotypes. It showed that the best basal salt
formulation differed among date palm genotypes and suggested culture stage-specific requirement.
Key words: Callus growth, In vitro, optimum medium, palm micropropagation, Phoenix dactylifera,
plant tissue culture, plant regeneration, somatic embryogenesis, commercial cultivars,
vegetative propagation
INTRODUCTION
Date palm (Phoenix dactylifera L.), a
monocotyledonous, dioecious tree species belonging to
Arecaceae family, is a major agricultural crop
cultivated in the hot, arid regions of the world, mainly
in the Middle East and North Africa. It contributes to a
sustainable agricultural system in areas where it is
grown and plays important social and economic roles
(Zohary and Hopf, 2000). Propagation by seeds
produce off-type plants and vegetative propagation by
offshoots has numerous limitations related to
availability and field survivability. Alternatively,
micropropagation has gained acceptance for large-
scale propagation and commercial production of date
palm trees. In addition, it offers controlled conditions
for studying aspects of physiological and genetic
improvement.
Micropropagation of date palm has been achieved
from several genotypes through organogenesis and
somatic embryogenesis using various meristematic
explants including zygotic embryos, shoot tips and
lateral buds (Omar et al., 1992; Al-Khayri, 2005; 2007).
Somatic embryogenesis is considered as the most
efficient regeneration process for date palm
micropropagation (El Hadrami and Baziz, 1995; Al-
Khayri, 2001; Fki et al., 2003). Several studies were
conducted to optimize somatic embryogeneis of date
palm through the manipulation of culture medium
ingredients including amino acids (Abo El-Nil, 1989a;
Abdel-Rahim et al., 1998; El-Shiaty et al., 2004;
Zouine and El Hadrami, 2007), plant growth regulators