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The study of development and in vitro multiplication of Actinidia chinensis Planch


Author: Romanciuc Gabriela
Degree:doctor of biology
Speciality: 03.00.05 - Botany
Year:2009
Scientific adviser: Alexandru Ciubotaru
doctor habilitat, professor, Botanical Garden (Institute)
Institution:
Scientific council:

Status

The thesis was presented on the 11 August, 2009
Approved by NCAA on the 1 October, 2009

Abstract

Adobe PDF document – 0.49 Mb / in romanian

Keywords

actinidia chinensis, in vitro culture, medium, plant hormones, callus, meristematic centers, organogenesis, regenerated plants

Summary

This work contains the results of scientific researches of in vitro plant regeneration of Actinidia chinensis Planch. The aim of this study was the establishment of the optimal relationship between culture medium, growth regulators and culture conditions for all stages of micropropagation of mentioned specie.

An efficient plant regeneration protocol for Actinidia chinensis was developed via organogenesis from the leaf blades and petioles. The explants were cultured on different medium such as S 2,5 (Standardi A. 1983), B5 (Gamborg, 1968), MS (Murashige-Skoog, 1962) with different concentrations of exogenous plant hormones as BAP (benzyladenine), Zeatin, IBA (indole-3-butyric acid), ANA (α-naphthaleneacetic acid), 2,4-D (2,4-dichlorophenoxyacetic acid) and GA3 (gibberellic acid). The first seven days the explants were kept in the dark and then transferred in the culture room. The cultures were incubated at a temperature of 24±1°C under 16-h photoperiod with illumination by white fluorescent light with an intensity of 2000. The callus initiation started in about two weeks after inoculation. Optimal callus induction was established for S 2,5 medium with 0,02mg/l ANA and 1,0 mg/l Zeatin for all types of explants. Was distinguished two types of callus present in culture: morphogenetic and nonmorphogenic, which differed not only in their capacity for organ formation, but also in their structure, appearance and morphology. Histological studies of calli segments confirmed this fact after 4-5 month of cultivation. The callus was subcultured into regeneration medium. This medium was S 2,5 added with 1,0 mg/l BAP and 1,0 mg/l GA3 which was established more effective for shoot formation and plant regeneration, especially for Actinidia culture. Histological examination revealed the presence of meristematic centers in morphologic callus after one month of culture on regeneration medium. These centers consisted of meristematic cells with regeneration ability. Cells with numerous starch grains were presented adjacent to these centers. Transferring regenerated shoots without roots to new rooting medium produced whole fertile plants.