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Method for cultivating passionflower sterile explant

A cultivation method and explant technology are applied in the cultivation field of Passiflora aseptic explants, which can solve problems such as decreased survival rate, endophyte pollution, and difficult eradication of endophyte, so as to reduce production costs and Pollution rate, effect of reducing browning rate and mortality

Inactive Publication Date: 2021-11-05
YUNNAN INST OF TROPICAL CROPS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But it still has the following disadvantages: 1. With the increase of disinfectant concentration or disinfection time, the pollution rate is in a downward trend, while the mortality rate and browning rate are in an upward trend, and the survival rate is in a downward trend
2. Endophytes are difficult to eradicate. With the increase of the number of subcultures, the pollution of endophytes will also become serious

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Take the explants at 10:00 in the morning, select the terminal buds and 3 axillary buds, about 13.5cm long, and the freshly drawn tender stems of passionflower with 1 leaf on the top of the terminal buds as explants, soak them in saturated washing powder liquid After 20 minutes, the explants were rinsed with tap water, and then rinsed with sterile water for 3 times. Using a rifampicin solution with a concentration of 100 mg / L, the explants were immersed in the rifampicin solution for 2.5 cm, and pre-cultured for 3 days at a temperature of 26°C and a light time of 10 h / d to obtain pre-cultured explants . The precultured explants were cut to grow into 1.5 cm stem segments with 1 axillary bud. The stems were sterilized with 75% alcohol by volume for 30 seconds, then surface-sterilized with 0.1% mercury chloride for 11 minutes, and washed 4 times to obtain aseptic stems. Then inoculate each aseptic stem section in the 27ml MS basic medium without adding any hormones, cult...

Embodiment 2

[0038] Take explants at 12 o'clock in the morning, select terminal buds and 3 axillary buds, about 12cm in length, and the freshly pumped young stems of passionflower with 1 leaf on the top of the terminal buds as explants, soak the explants with saturated washing powder liquid After 10 minutes of implantation, rinse in tap water, and then rinse twice with sterile water. Using a rifampin solution with a concentration of 80 mg / L, the explants were immersed in the rifampicin solution for 2 cm, and pre-cultured for 2 days at a temperature of 22° C. and a light time of 8 h / d to obtain pre-cultured explants. The precultured explants were cut to grow into 0.8 cm stem segments with 1 axillary bud. The stems were sterilized with 75% alcohol by volume for 10 seconds, then surface-sterilized with 0.05% mercury chloride for 10 minutes, and washed three times to obtain aseptic stems. Then inoculate each aseptic stem section in the 25ml MS basic medium without adding any hormones, cultiva...

Embodiment 3

[0040]Take the explants at 16:00 in the afternoon, select the terminal buds and 4 axillary buds, about 15cm long, and the freshly drawn tender stems of passionflower with 2 leaves on the top of the terminal buds as explants, soak the explants with saturated washing powder After 30 minutes of implantation, rinse with tap water, and then rinse with sterile water for 4 times. Using a rifampin solution with a concentration of 120 mg / L, the explants were immersed in the rifampicin solution for 3 cm, and pre-cultured for 4 days at a temperature of 30° C. and a light time of 12 h / d to obtain pre-cultured explants. The precultured explants were cut to grow to 2.0 cm stem segments with 2 axillary buds. The stems were sterilized with 90% alcohol by volume for 60 seconds, then surface-sterilized with 0.15% mercury chloride for 12 minutes, and washed 5 times to obtain aseptic stems. Then inoculate each aseptic stem segment in 30ml MS basic medium without adding any hormones, and cultivat...

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Abstract

The invention provides a method for cultivating a passionflower sterile explant, and relates to the technical field of plant tissue culture. The method comprises the following steps: collecting a passionflower tender stem segment as an explant, washing the explant, then soaking the explant in a saturated washing powder solution, and rinsing the explant to obtain a pretreated explant; carrying out sterile pre-culture on the pretreated explant by adopting a rifampicin solution to obtain a pre-cultured explant; cutting the pre-cultured explant into stem segments, and disinfecting the stem segments to obtain sterile stem segments; and culturing the sterile stem segments in an MS basic culture medium to obtain the sterile passionflower explant. According to the cultivation method disclosed by the invention, by combining a two-time pretreatment method of the explant, a mercury bichloride sterilization method, time and the like, the pollution rate is reduced, and meanwhile, the browning rate and the death rate caused by disinfection and sterilization are also reduced, so that the survival rate of the explant is greatly increased, and the production cost of tissue culture is reduced.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for cultivating Passiflora aseptic explants. Background technique [0002] Passiflora (Passifloracaerulea L.) is a typical tropical and subtropical berry fruit tree, belonging to the Passifloraceae (Passifloraceae) genus Passiflora (Passiflora L.), and is a perennial evergreen vine. Passion fruit has a unique and strong aromatic smell, sweet and sour taste, is a famous tropical fruit for juice, and has the reputation of "king of juice". The tissue culture technique of Passiflora is so far difficult to be applied to seedling breeding, and one of the main reasons is that the explants are difficult to sterilize, especially from the explants collected in the field, because the surface and the inside all carry a certain amount of bacteria and fungi, especially endophytes. The current research on the tissue culture technology of Passiflora takes the stems of Pas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/008A01H4/002
Inventor 管艳李玲刘世红殷振华桂明春粱国平唐敏田海
Owner YUNNAN INST OF TROPICAL CROPS
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