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In vitro culture directed towards plant improvement of tea (Camellia sinensis var.assamica)

In vitro culture directed towards plant improvement of tea (Camellia sinensis var.assamica)
In vitro culture directed towards plant improvement of tea (Camellia sinensis var.assamica)

In vitro culture of tissues of tea Camellia sinensis var. assamica were undertaken with the aim of developing protocols for use in plant improvement. As a step towards the development of a micropropagation system for tea, shoots were recovered from nodal explants through enhanced axillary bud formation. Shoot cultures were maintained and multiplied by dividing into nodal segments. These shoots were successfully rooted by pre-treating with a concentrated IBA solution followed by culture in auxin-free half-strength MS liquid medium. Rooted plantlets were established in a soil mixture and were weaned ex vitro.

Explants from cotyledon, stem and leaf were tested for their morphogenic potential in vitro. Depending on the culture conditions, explants showed different morphogenic capacities, and morphogenesis occurred through different developmental pathways. In cotyledon explants the presence of 2,4-D, in contrast to NAA, seems necessary for the induction of embryogenic cells. Somatic embryo development occurred without prolific callus formation with subsequent removal of 2,4-D in the medium. Histological studies were carried out to examine the position and activity of competent cells leading to somatic embryo formation. Indirect shoot organogenesis was achieved via callus formed in stem tissue with gradual alteration of growth regulators in the medium over time during callus sub-culture. According to the histological observations, in stem explant tissues, cell division occurred in pith parenchyma cells and these developed to form callus. Leaf tissues produced only callus or in some instances roots together with callus.

Protoplasts were isolated from leaf mesophyll tissue of tea using various enzyme combinations and concentrations. The various factors which affect protoplast yield and viability were assessed to obtain optimum protoplast yield and viability. Preliminary studies were also carried out to identify the requirement for culture of these isolated protoplasts.

University of Southampton
Gunasekare, Malliyawadu Trixie Kumudini
Gunasekare, Malliyawadu Trixie Kumudini

Gunasekare, Malliyawadu Trixie Kumudini (1997) In vitro culture directed towards plant improvement of tea (Camellia sinensis var.assamica). University of Southampton, Doctoral Thesis.

Record type: Thesis (Doctoral)

Abstract

In vitro culture of tissues of tea Camellia sinensis var. assamica were undertaken with the aim of developing protocols for use in plant improvement. As a step towards the development of a micropropagation system for tea, shoots were recovered from nodal explants through enhanced axillary bud formation. Shoot cultures were maintained and multiplied by dividing into nodal segments. These shoots were successfully rooted by pre-treating with a concentrated IBA solution followed by culture in auxin-free half-strength MS liquid medium. Rooted plantlets were established in a soil mixture and were weaned ex vitro.

Explants from cotyledon, stem and leaf were tested for their morphogenic potential in vitro. Depending on the culture conditions, explants showed different morphogenic capacities, and morphogenesis occurred through different developmental pathways. In cotyledon explants the presence of 2,4-D, in contrast to NAA, seems necessary for the induction of embryogenic cells. Somatic embryo development occurred without prolific callus formation with subsequent removal of 2,4-D in the medium. Histological studies were carried out to examine the position and activity of competent cells leading to somatic embryo formation. Indirect shoot organogenesis was achieved via callus formed in stem tissue with gradual alteration of growth regulators in the medium over time during callus sub-culture. According to the histological observations, in stem explant tissues, cell division occurred in pith parenchyma cells and these developed to form callus. Leaf tissues produced only callus or in some instances roots together with callus.

Protoplasts were isolated from leaf mesophyll tissue of tea using various enzyme combinations and concentrations. The various factors which affect protoplast yield and viability were assessed to obtain optimum protoplast yield and viability. Preliminary studies were also carried out to identify the requirement for culture of these isolated protoplasts.

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Published date: 1997

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Local EPrints ID: 463073
URI: http://eprints.soton.ac.uk/id/eprint/463073
PURE UUID: d5aa5e79-c3d6-4015-b67d-08cf94d70424

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Date deposited: 04 Jul 2022 20:43
Last modified: 04 Jul 2022 20:43

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Author: Malliyawadu Trixie Kumudini Gunasekare

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