Anther separation method suitable for stevia rebaudianum anther culture

Abstract

The invention discloses an anther separation method suitable for stevia rebaudianum anther culture, and belongs to the technical field of plant tissue culture. The method mainly comprises the following steps: collecting capitula with a suitable size in the bud stage of stevia rebaudianum, disinfecting the capitula with 75% ethanol under an aseptic condition for 30 s, immersing the disinfected capitula in 1% mercuric chloride for 3-4 min, cleaning the immersed capitula with sterile water 3-4 times, adding the capitula into a sterile filter paper paved culture dish, airing the capitula, selecting buds from the capitula by using tweezers, placing the selected capitula on the finger pulp of a left index finger, piercing the tops of the corollas of the buds by a dissecting needle held by a right hand, and carrying out rolling extrusion on the corollas from the basal part to the top by using the dissecting needle in order to extrude anthers which are complete and directly used for in vitro culture. The method for separating the anthers from the minimal buds of stevia rebaudianum, with the length of about 2 mm, is disclosed in the invention, and has the advantages of simplicity, rapidness, maintenance of the completeness of the anthers, increase of the separation success rate, and effective reduction of the pollution in the anther culture.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for isolating stevia (Steviarebaudiana Bertoni) plant anthers, which can be used for the research on in vitro culture of stevia anthers to induce haploid plants. Background technique [0002] Stevia (Stevia rebaudiana Bertoni) is an emerging natural high-sweetness and low-calorie sugar source plant. The research on its germplasm resources and the cultivation of new fine varieties are very important to the development of stevia industry. Because Stevia has the characteristics of cross-pollination and self-incompatibility, its self-seed setting rate is extremely low, and due to the complex genetic background, the population traits of hybrid offspring are significantly separated, and it is difficult to directly obtain inbred lines and homozygous plants. Slowed down the stevia breeding process. [0003] Inducing plant megaspores and microspores to o...

AI Technical Summary

Problems solved by technology

Because Stevia has the characteristics of cross-pollination and self-incompatibility, its self-seed setting rate is extremely low, and due to the complex genetic background, the population traits of hybrid offspring are significantly separated, and it is difficult to directly obtain inbred lines and homozygous plants. Slowed Stevia Breeding Progress

[0003] Inducing plant megaspores and microspores to obtain haploid plants by tissue culture and other methods is an effective method to obtain homozygous plants. It has been successful in many plants such as corn, wheat, and cabbage. However, there are few studies on stevia. No success reported so far

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