Plant regeneration method by using giant tea rose stem segment as explant

Abstract

The invention provides a plant regeneration method by using a giant tea rose stem segment as an explant. The method provided by the invention comprises the following steps of: cutting the central section of a giant tea rose branch into a segment, sterilizing, carrying out subculture for one month, transversely cutting giant tea rose aseptic seedling blades, inoculating to a callus induction medium until embryogenic callus grows, culturing in a differentiation medium until the embryogenic callus differentiates adventitious buds, inoculating to a strong seedling medium, and inoculating to a rooting medium after growing tall so as to form the complete plant. According to the invention, the explant used is convenient to collect such that propagation of giant tea rose and cell culture can be carried out every year; the obtained embryogenic callus can be used as a good acceptor for genetic transformation, thus providing the acceptor for the establishment of a giant tea rose genetic transformation system; and the established regeneration system can help solve giant tea rose breeding problems so as to better protect and utilize the resource.

Description

[0001] technical field

[0002] The invention relates to a regeneration method of a plant, in particular to a method for regenerating a rose flower plant through an organogenesis pathway using a single-bud stem section of the rose flower perfume as an explant, and belongs to the technical field of plant tissue culture. Background technique

[0003] R.odorata var. gigantea is a deciduous vine of Rosaceae (Rosaceae). It is a single flower, milky white, fragrant, and the flower diameter is 8-10cm. The place of origin is Yunnan. It is one of the important original parents of modern roses, an important provenance in the evolution of roses, and a germplasm resource for cultivating large-flowered varieties and large-plant roses. When using traditional hybrid breeding methods to breed new varieties, not only the cycle is long and the workload is heavy, but also the problem of not fruiting will occur when hybridizing with other wild species or varieties; using the cuttings of the big...

Examples

Embodiment 1

[0026] A. Select the middle branches of the Chinese rose with full axillary buds, cut them into single-bud stems, sterilize them with 0.1% mercuric chloride for 8 minutes, rinse them with sterile water, and inoculate them in 1.5mg / L 6-BA + 0.15 mg / L NAA + 6.2g / L agar + 30g / L sucrose, and the MS basic medium with a pH value of 6.0, until the axillary buds germinated, and then inoculated on 1.0mg / L 6-BA + 0.01mg / L On the MS basic medium of NAA+6.2g / L agar+30g / L sucrose, under the conditions of 25°C, light intensity 2000Lx, and light time 14h / d, subculture was carried out for 25 days to obtain the free Bacteria;

[0027] B. Cut off the leaves of the aseptic seedlings of the large flower perfume rose obtained in step A, cut the wound transversely at the main vein of the leaflet, and inoculate the leaf back on the following callus induction medium: MS basic medium + 8 mg / L 2 , 4-D + 30g / L glucose + 2.5g / L phytogel, the pH value is 6.0, and cultured under dark conditions, the inc...

Embodiment 2

[0031] Steps A, C, and D are the same as in Example 1, except that the amount of 2,4-D in step B is 6 mg / L.

Embodiment 3

[0033] Steps A, C, and D are the same as in Example 1, except that the amount of 2,4-D in step B is 5 mg / L.

[0001] technical field

[0002] The invention relates to a regeneration method of a plant, in particular to a method for regenerating a rose flower plant through an organogenesis pathway using a single-bud stem section of the rose flower perfume as an explant, and belongs to the technical field of plant tissue culture. Background technique

[0003] R.odorata var. gigantea is a deciduous vine of Rosaceae (Rosaceae). It is a single flower, milky white, fragrant, and the flower diameter is 8-10cm. The place of origin is Yunnan. It is one of the important original parents of modern roses, an important provenance in the evolution of roses, and a germplasm resource for cultivating large-flowered varieties and large-plant roses. When using traditional hybrid breeding methods to breed new varieties, not only the cycle is long and the workload is heavy, but also the problem of not fruiting will occur when hybridizing with other wild species or varieties; using the cuttings of the big...

Examples

Embodiment 1

[0026] A. Select the middle branches of the Chinese rose with full axillary buds, cut them into single-bud stems, sterilize them with 0.1% mercuric chloride for 8 minutes, rinse them with sterile water, and inoculate them in 1.5mg / L 6-BA + 0.15 mg / L NAA + 6.2g / L agar + 30g / L sucrose, and the MS basic medium with a pH value of 6.0, until the axillary buds germinated, and then inoculated on 1.0mg / L 6-BA + 0.01mg / L On the MS basic medium of NAA+6.2g / L agar+30g / L sucrose, under the conditions of 25°C, light intensity 2000Lx, and light time 14h / d, subculture was carried out for 25 days to obtain the free Bacteria;

[0027] B. Cut off the leaves of the aseptic seedlings of the large flower perfume rose obtained in step A, cut the wound transversely at the main vein of the leaflet, and inoculate the leaf back on the following callus induction medium: MS basic medium + 8 mg / L 2 , 4-D + 30g / L glucose + 2.5g / L phytogel, the pH value is 6.0, and cultured under dark conditions, the inc...

Embodiment 2

[0031] Steps A, C, and D are the same as in Example 1, except that the amount of 2,4-D in step B is 6 mg / L.

Embodiment 3

[0033] Steps A, C, and D are the same as in Example 1, except that the amount of 2,4-D in step B is 5 mg / L.

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