Strosse et al. (2006) stated that the efficiency of a plant
regeneration system through cell suspension cultures in banana
is determined by the following key factors: (1) the
availability of embryogenic-competent explants, (2) the nature
and frequency of embryogenic response, (3) the success
rate of suspension initiation, (4) the long time and continuous
effort to generate and improve the quality of the cell suspension,
and (5) the plant regeneration frequency. In this work,
we could overcome most of these bottlenecks and were
successful in developing a simple and efficient somatic embryogenesis
system. According to previous reports the establishment
of the embryogenic cell suspension is strongly
influenced by genotype; culture initiation is difficult and can
take anywhere from 6 to 12 mo (Strosse et al. 2006). Obviously,
banana cell suspensions obtained with great difficulty
are expected to be useful for a proportionately longer time
period. Maintaining the embryogenic cell suspensions, in
general, also is a laborious and time-consuming task due to
frequent subculture requirements that may lead to an increase
in the frequency of somaclonal variation, microbial
contamination, and eventually the complete loss of morphogenetic
potential (Kulkarni and Ganapathi 2009). Expensive
cryopreservation, therefore, is considered an alternative for
the safe storage of established valuable cell suspension cultures.
Laboratories are also using slow-growth maintenance
techniques to maintain suspension cultures (Kulkarni and
Ganapathi 2009), which requires expertise and involves cost.
In the current protocol, generation of suspension cultures
occurred within a short period compared to previous reports
(Fig. 2). Therefore, the protocol described here reduces the
time required for generation of cell suspension from 14 to
42 mo to just 3 mo, and as the technique is very simple, it has
the potential to be used in any laboratory engaged in genetic
improvement of banana
Strosse et al. (2006) stated that the efficiency of a plantregeneration system through cell suspension cultures in bananais determined by the following key factors: (1) theavailability of embryogenic-competent explants, (2) the natureand frequency of embryogenic response, (3) the successrate of suspension initiation, (4) the long time and continuouseffort to generate and improve the quality of the cell suspension,and (5) the plant regeneration frequency. In this work,we could overcome most of these bottlenecks and weresuccessful in developing a simple and efficient somatic embryogenesissystem. According to previous reports the establishmentof the embryogenic cell suspension is stronglyinfluenced by genotype; culture initiation is difficult and cantake anywhere from 6 to 12 mo (Strosse et al. 2006). Obviously,banana cell suspensions obtained with great difficultyare expected to be useful for a proportionately longer timeperiod. Maintaining the embryogenic cell suspensions, ingeneral, also is a laborious and time-consuming task due tofrequent subculture requirements that may lead to an increasein the frequency of somaclonal variation, microbialcontamination, and eventually the complete loss of morphogeneticpotential (Kulkarni and Ganapathi 2009). Expensivecryopreservation, therefore, is considered an alternative forthe safe storage of established valuable cell suspension cultures.Laboratories are also using slow-growth maintenancetechniques to maintain suspension cultures (Kulkarni andGanapathi 2009), which requires expertise and involves cost.In the current protocol, generation of suspension culturesoccurred within a short period compared to previous reports(Fig. 2). Therefore, the protocol described here reduces thetime required for generation of cell suspension from 14 to42 mo to just 3 mo, and as the technique is very simple, it hasthe potential to be used in any laboratory engaged in geneticimprovement of banana
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