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A temporary immersion system improves in vitro regeneration of peach palm through secondary somatic embryogenesis

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Background and Aims Secondary somatic embryogenesis has been postulated to occur during induction of
peach palm somatic embryogenesis. In the present study this morphogenetic pathway is described and a protocol for the establishment of cycling cultures using a temporary immersion system (TIS) is presented.
Methods Zygotic embryos were used as explants, and induction of somatic embryogenesis and plantlet growth were compared in TIS and solid culture medium. Light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to describe in vitro morphogenesis and accompany morpho-histological alterations during culture.
Key Results The development of secondary somatic embryos occurs early during the induction of primary
somatic embryos. Secondary somatic embryos were observed to develop continually in culture, resulting in
non-synchronized development of these somatic embryos. Using these somatic embryos as explants allowed development of cycling cultures. Somatic embryos had high embryogenic potential (65.8+3.0 to 86.2+5.0%) over the period tested. The use of a TIS greatly improved the number of somatic embryos obtained, as well as subsequent plantlet growth. Histological analyses showed that starch accumulation precedes the development of somatic embryos, and that these cells presented high nucleus/cytoplasm ratios and high mitotic indices, as evidenced by DAPI staining. Morphological and SEM observations revealed clusters of somatic embryos on one part of the explants, while other parts grew further, resulting in callus tissue. A multicellular origin of the secondary somatic embryos is hypothesized. Cells in the vicinity of callus accumulated large amounts of phenolic substances in their vacuoles. TEM revealed that these cells are metabolically very active, with the presence of numerous mitochondria and Golgi apparatuses. Light microscopy and TEM of the embryogenic sector revealed cells with numerous amyloplasts, large nuclei and nucleoli, and numerous plasmodesmata. Plantlets were obtained and after 3 months in culture their growth was significantly better in TIS than on solid culture medium. However, during acclimatization the survival rate of TIS-grown plantlets was lower.
Conclusions The present study confirms the occurrence of secondary somatic embryos in peach palm and
describes a feasible protocol for regeneration of peach palm in vitro. Further optimizations include the use of explants obtained from adult palms and improvement of somatic embryo conversion rates.
Key words: Bactris gasipaes, tissue culture, somatic embryogenesis, clonal propagation, Picloram.

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Informasi Detail

Judul Seri

Annals of Botany

No. Panggil

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Penerbit

: Oxford University Press on behalf of the Annals of Botany Company., 2011

Deskripsi Fisik

Annals of Botany 108: 1463–1475, 2011

Bahasa

English

ISBN/ISSN

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Klasifikasi

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Tipe Isi

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Tipe Media

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Tipe Pembawa

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Edisi

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Subjek

PERTANIAN.

Info Detail Spesifik

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Pernyataan Tanggungjawab

yuli/agus

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