One-step tissue culture and rapid propagation method of grapes

Abstract

The invention discloses a one-step tissue culture and rapid propagation method of grapes. The method comprises the following steps of: S1, sterilizing explants, namely collecting current immature stems of semi-lignified grapes, chopping the immature stems into stems with buds, washing the stems with buds by running water, and sterilizing the stems with buds on a ultra-clean table by a combined sterilizing mode of alcohol and mercuric chloride; and S2, inoculating and propagating, namely inoculating the sterilized stems with buds on a culture medium to grow, and culturing an inoculated culture material in a culture room with illumination intensity of 3000LX and illumination time of 16h / d and at 24-26 DEG.C, so that after one week, the stems with buds grow into complete plants. The method has the beneficial effects that the one-step rapid propagation of simultaneously propagating and rooting the stems with buds of the grapes is realized, the propagation, strong seedling cultivation and rooting culture can be synchronously performed by only one culture medium, the tissue cultured seedlings can grow healthily, the production cost is low, and the obtained tissue culture seedlings are consistent in heredity, the propagation coefficient is high, the propagation period is short, the operation difficulty is reduced, and the culture efficiency is improved.

Description

technical field

[0001] The invention relates to a grape cultivation method, in particular to a one-step tissue culture and rapid propagation method for grapes. Background technique

[0002] Grape is a crop with wide application and high economic value. In terms of cultivation methods, there is a gradual transition from traditional self-rooted seedlings to grafted seedlings. Conventional propagation mostly uses grafting and cuttings, but these two propagation methods are likely to cause virus accumulation, and the propagation time is limited by seasons. Tissue culture is widely used in the rapid propagation of grapes. Its advantages can not only speed up the propagation speed, but also combine heat treatment with shoot tip culture to effectively eliminate viruses and obtain non-toxic seedlings. The tissue culture and rapid propagation of grapes can effectively solve the technical problems of grape reproduction, and has been maturely applied in grape reproduction. However, ...

Examples

Embodiment 1

[0012] Grape one-step tissue culture rapid propagation method, it comprises the following steps:

[0013] S1. Sterilize the explants, collect the semi-lignified grape stems of the year, cut them into sections with buds, wash them with running water, and then sterilize them on an ultra-clean bench. The sterilization method is a combination of alcohol and mercuric chloride , first sterilize with alcohol for 45s, and then sterilize with mercuric chloride for 4min;

[0014] S2, inoculation and propagation, inoculate the sprouted stem section after sterilization on the growth medium, such as figure 1 As shown, the medium is MS+NAA0.2mg / L+IAA0.1mg / L, and the inoculated culture material is placed in a culture with a light intensity of 3000LX, a light time of 16h / d, and a temperature of 25°C. Chamber culture, the reproduction cycle is one week, to realize the simultaneous multiplication and rooting of the culture materials, and after one week, the budded stem segments grow into compl...

Embodiment 2

[0016] Grape one-step tissue culture rapid propagation method, it comprises the following steps:

[0017] S1. Sterilize the explants. Collect semi-lignified grape stems that were born in the same year, cut them into 2-3cm single-bud stem segments, wash them with running water, and then sterilize them on an ultra-clean bench. The sterilization method uses alcohol plus mercury chloride. Combined sterilization method, first rinse with 70% alcohol by volume for 30 seconds, then rinse with sterile water for 3 times, soak in 0.1% mercury solution for disinfection for 5 minutes, and rinse with sterile water for 3 to 5 minutes Second-rate,

[0018] S2, inoculation and propagation, inoculate the sprouted stem section after sterilization on the growth medium, such as figure 1 As shown, the medium is MS+NAA0.2mg / L+IAA0.1mg / L, pH 5.8-6.0, and the inoculated culture material is placed in a light intensity of 3000LX, a light time of 16h / d, and a temperature of It is cultivated in a cultur...

Embodiment 3

[0020] Grape one-step tissue culture rapid propagation method, it comprises the following steps:

[0021] S1. Sterilize the explants. Collect semi-lignified grape stems that were born in the same year, cut them into 2-3cm single-bud stem segments, wash them with running water, and then sterilize them on an ultra-clean bench. The sterilization method uses alcohol plus mercury chloride. Combined sterilization method, first rinse with 70% alcohol by volume for 60 seconds, then rinse with sterile water for 3 times, soak in 0.1% mercury solution for disinfection for 3 minutes, rinse with sterile water for 3 times,

[0022] S2, inoculation and propagation, inoculate the sprouted stem section after sterilization on the growth medium, such as figure 1 As shown, the medium is MS+NAA0.2mg / L+IAA0.1mg / L, pH 5.8, and the inoculated culture material is placed in a light intensity of 3000LX, a light time of 16h / d, and a temperature of 26 Cultivate in a culture room at ℃, and the propagation...

technical field

[0001] The invention relates to a grape cultivation method, in particular to a one-step tissue culture and rapid propagation method for grapes. Background technique

[0002] Grape is a crop with wide application and high economic value. In terms of cultivation methods, there is a gradual transition from traditional self-rooted seedlings to grafted seedlings. Conventional propagation mostly uses grafting and cuttings, but these two propagation methods are likely to cause virus accumulation, and the propagation time is limited by seasons. Tissue culture is widely used in the rapid propagation of grapes. Its advantages can not only speed up the propagation speed, but also combine heat treatment with shoot tip culture to effectively eliminate viruses and obtain non-toxic seedlings. The tissue culture and rapid propagation of grapes can effectively solve the technical problems of grape reproduction, and has been maturely applied in grape reproduction. However, ...

Examples

Embodiment 1

[0012] Grape one-step tissue culture rapid propagation method, it comprises the following steps:

[0013] S1. Sterilize the explants, collect the semi-lignified grape stems of the year, cut them into sections with buds, wash them with running water, and then sterilize them on an ultra-clean bench. The sterilization method is a combination of alcohol and mercuric chloride , first sterilize with alcohol for 45s, and then sterilize with mercuric chloride for 4min;

[0014] S2, inoculation and propagation, inoculate the sprouted stem section after sterilization on the growth medium, such as figure 1 As shown, the medium is MS+NAA0.2mg / L+IAA0.1mg / L, and the inoculated culture material is placed in a culture with a light intensity of 3000LX, a light time of 16h / d, and a temperature of 25°C. Chamber culture, the reproduction cycle is one week, to realize the simultaneous multiplication and rooting of the culture materials, and after one week, the budded stem segments grow into compl...

Embodiment 2

[0016] Grape one-step tissue culture rapid propagation method, it comprises the following steps:

[0017] S1. Sterilize the explants. Collect semi-lignified grape stems that were born in the same year, cut them into 2-3cm single-bud stem segments, wash them with running water, and then sterilize them on an ultra-clean bench. The sterilization method uses alcohol plus mercury chloride. Combined sterilization method, first rinse with 70% alcohol by volume for 30 seconds, then rinse with sterile water for 3 times, soak in 0.1% mercury solution for disinfection for 5 minutes, and rinse with sterile water for 3 to 5 minutes Second-rate,

[0018] S2, inoculation and propagation, inoculate the sprouted stem section after sterilization on the growth medium, such as figure 1 As shown, the medium is MS+NAA0.2mg / L+IAA0.1mg / L, pH 5.8-6.0, and the inoculated culture material is placed in a light intensity of 3000LX, a light time of 16h / d, and a temperature of It is cultivated in a cultur...

Embodiment 3

[0020] Grape one-step tissue culture rapid propagation method, it comprises the following steps:

[0021] S1. Sterilize the explants. Collect semi-lignified grape stems that were born in the same year, cut them into 2-3cm single-bud stem segments, wash them with running water, and then sterilize them on an ultra-clean bench. The sterilization method uses alcohol plus mercury chloride. Combined sterilization method, first rinse with 70% alcohol by volume for 60 seconds, then rinse with sterile water for 3 times, soak in 0.1% mercury solution for disinfection for 3 minutes, rinse with sterile water for 3 times,

[0022] S2, inoculation and propagation, inoculate the sprouted stem section after sterilization on the growth medium, such as figure 1 As shown, the medium is MS+NAA0.2mg / L+IAA0.1mg / L, pH 5.8, and the inoculated culture material is placed in a light intensity of 3000LX, a light time of 16h / d, and a temperature of 26 Cultivate in a culture room at ℃, and the propagation...

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