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Control method of plant tissue culture seedling endophytes

A technology of plant tissue culture and control method, which is applied in botany equipment and methods, plant regeneration, horticultural methods, etc. It can solve problems such as easy pyrolysis, complicated process, and drug resistance, and achieves remarkable effects, simple operation, low cost effect

Inactive Publication Date: 2020-09-11
江苏丰收大地种业发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But this method is cumbersome and it is difficult to eliminate the possibility of endophyte contamination; 2. Add antibiotics or bactericides to the culture medium, but the current effect is not very satisfactory
The reason is that antibiotics can only inhibit the growth of some bacteria, and are easy to decompose at high temperature. They need to be added after high-temperature disinfection. The process is complicated, and long-term use will produce drug resistance, and the effect is difficult to reduce the pollution to an ideal state.

Method used

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  • Control method of plant tissue culture seedling endophytes

Examples

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Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: as figure 1 As shown, the control method of the endophytic bacteria in the plant tissue culture seedlings specifically comprises the following steps:

[0025] (1) Take explants: under the sunny and dry weather in spring and autumn, the temperature is 15-25° C., and the relative humidity is lower than 60%. The explants are garlic, and the plants are transplanted before taking the explants. In the flowerpot, be placed in the greenhouse (temperature is controlled above 25 ℃) to grow, and spray broad-spectrum fungicide, initially reduce the quantity of surface microorganism and plant endophyte; The method of taking described explant is: Rinse the garlic seedlings that have been sprayed with 50% carbendazim wettable powder 300 times; solution disinfection in tap water for 2 hours, and use a sterile knife to peel off the scales of the buds in a sterile environment, and cut them below the growth point. Cut off at 0.5-1cm to form explants with growth points, and ...

Embodiment 2

[0032] Embodiment 2: the control method of endophytic bacteria of this plant tissue culture seedling, specifically comprises the following steps:

[0033] (1) Take explants: in the sunny and dry weather in spring and autumn, the temperature is at 15-25° C., and the relative humidity is lower than 60%. Take explants, and the explants are sweet potatoes. Transplant the plants into flowerpots, place them in the greenhouse to grow at a temperature above 25°C, and spray broad-spectrum fungicides to initially reduce the number of surface microorganisms and plant endophytes. The method of using explants is: The sweet potato seedlings on the sterilized sweet potato tubers are cut off at 0.5-1cm below the growth point with a sterile knife or scissors or tweezers, and the stem segment with the growth point is the explant, which is reserved for later use. ;

[0034] (2) Surface disinfection of explants: the explants taken were rinsed in running water for 2 hours, then rinsed twice with ...

Embodiment 3

[0040] Embodiment 3: the control method of endophytic bacteria in tissue culture seedlings of Nantian bamboo specifically comprises the following steps:

[0041] (1) Take explants: in the sunny and dry weather in spring and autumn, the temperature is at 15-25°C, and the relative humidity is lower than 60%. Transplant into flowerpots, grow in a greenhouse above 25°C, and spray the broad-spectrum fungicide carbendazim to initially reduce the number of surface microorganisms and plant endophytes; in sunny and dry weather in spring and autumn Take explants, the method of taking explants is: cut off the tender shoots of Nantian bamboo, the length of the bud segment is 1-2cm, and the bud segment with the growth point is the explant; set aside;

[0042] The method of explant described in described step (1) is:

[0043] (2) Surface disinfection of explants: the explants taken were rinsed in running water for 2 hours, then rinsed twice with sterile water, then soaked in 75% alcohol fo...

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Abstract

The invention discloses a control method of plant tissue culture seedling endophytes. The control method specifically comprises the following steps: (1) collecting explants for later use; (2) disinfecting the surfaces of the explants: flushing the collected explants with running water for 2 hours, and sequentially soaking the explants into alcohol, an infiltrative disinfectant A solution and HgClfor disinfection; (3) carrying out inoculation and induced culture on the explants: preparing an explant induced culture medium, adding a sterilizing agent into the explant induced culture medium, inoculating the explants into a disinfected culture medium flask containing the explant induced culture medium, and carrying out induced growing point culture in a sterile culture room under a lighting condition, so as to obtain seedlings; (4) carrying out subculture: after the newborn seedlings grow to a length of 3cm or above, carrying out second-generation subculture by virtue of the explant induced culture medium in the step (3); and (5) acquiring the plant tissue culture seedling endophytes: after the second-generation subculture is finished, selecting tissue culture seedlings which healthyand strongly and are not polluted by any bacteria and fungus, namely the seedlings without endophyte.

Description

technical field [0001] The invention relates to a method for controlling bacteria in the process of plant tissue culture, in particular to a method for controlling endophytic bacteria in plant tissue culture seedlings. Background technique [0002] Tissue culture technology is the most important technology to solve the problem of crop detoxification and rejuvenation and rapid seedling production. Endophytic bacteria in plants are bacteria that live in the intercellular space or in cells of various tissues and organs of healthy plants. Since endophytic bacteria cannot be removed by general surface disinfection methods, they will be brought into the culture process along with the explants, and will be in the tissue. Obvious or inconspicuous bacterial colonies appear continuously during the cultivation process, which will cause large-scale contamination of the medium during tissue culture of some plants, resulting in culture failure. Therefore, the method of controlling endoph...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/00A01H4/001A01H4/008
Inventor 周志疆陈雯雯杨斌相鸿雁杨佩娟庄锦贵张桢
Owner 江苏丰收大地种业发展有限公司
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