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Method for screening root media by using tobacco sterile seedlings

A technology of rooting medium and tobacco seeds, which is applied in the field of plant tissue culture, can solve problems such as complex procedures and long cycles, and achieve the effects of simple process, speeding up the test progress, and saving manpower

Inactive Publication Date: 2013-04-24
四川省烟草技术中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the screening of the rooting medium of tobacco tissue culture seedlings mainly selects the adventitious buds induced by callus as the rooting identification material. Screening can only be carried out when identification materials are obtained at the first stage, and there are disadvantages such as long cycle and complicated procedures

Method used

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  • Method for screening root media by using tobacco sterile seedlings
  • Method for screening root media by using tobacco sterile seedlings
  • Method for screening root media by using tobacco sterile seedlings

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment one, as figure 1 Shown is a schematic flow chart of the screening method of the present invention.

[0035] Wherein, the method for screening the rooting medium of tobacco sterile seedlings is as follows:

[0036] A, the step of disinfecting tobacco seeds;

[0037] B, the step of cultivating sterile seedlings;

[0038] C, the step of cutting sterile seedlings:

[0039] D, configure some steps of rooting medium to be selected;

[0040] E, the step of screening optimal rooting medium.

[0041] Step A is specifically as follows: select 150 uncoated tobacco seeds with full grains and place them in a petri dish, (choose pure leaf yellow tobacco seeds) add ethanol with a mass percentage concentration of 75% to soak for 30 seconds, and rinse with sterile water for 3 times; Then use the mass percent concentration as 0.1% HgCl 2 The solution was soaked for 7 minutes, and then washed 5 times with sterile water to obtain sterilized tobacco seeds.

[0042] Step B ...

Embodiment 2

[0055] Embodiment two, screening method is as follows

[0056] a. Select 140 uncoated Beinhart1000-1 tobacco seeds with full grains and place them in a petri dish, soak them in 75% ethanol for 30 seconds, rinse them with sterile water for 3 times; Rinse 5 times to obtain sterilized tobacco seeds.

[0057] b. Use sterile tweezers to inoculate the sterilized tobacco seeds in a petri dish containing 2% sucrose, 0.4% agar, and pH6.0 medium, the distance between seeds is 0.7cm, and the distance between the upper surface of the medium in the petri dish and the upper cover of the petri dish The distance should be controlled at 1.5cm, and the culture medium should be sterilized by damp heat at 121°C for 20 minutes; finally, the inoculated culture dish should be placed in a sterile environment at 28°C, 12h light / d, and cultivated for 10d; when the two cotyledons of the seedlings touch Cover the petri dish.

[0058] Taking the configuration of 1L of the above medium as an example, per...

Embodiment 3

[0068] Comparative test (1), method: explant-callus induction of adventitious buds-adventitious shoots rooting-seedling formation.

[0069] Tobacco cloud 85 seeds with full grains were selected, treated with 70% ethanol for 30 seconds, rinsed with sterile water for 3 times, then soaked in 0.1% HgCl2 solution for 7-9 minutes, rinsed with sterile water for 4-5 times, and inoculated in the seedlings with 2% sucrose, Germinate on 0.6% agar, hormone-free MS basic medium, and place in a constant temperature incubator at 25±0.5°C. Sterile seedlings of suitable seedling age were used as test materials.

[0070] Callus induction differentiation culture takes 2 / 3MS as basic medium, additional sucrose 30g / L, agar 5.5g / L, hormone: 6-BA0.5mg / L+NAA0.2mg / L, medium pH adjusted to 5.8, Sterilize at a constant temperature of 121-126°C for 20 minutes. When the aseptic seedlings grow to four leaves and one heart, take the shoot tips of the plants and inoculate them on the corresponding medium re...

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Abstract

The invention discloses a method for screening root media by using tobacco sterile seedlings. The method comprises the following steps: A, sterilizing tobacco seeds; B, cultivating the sterile seedlings; C, shearing the sterile seedlings; D, allocating a plurality of root media to be screened; and E, screening the optimal root media. The method is simple in process, labor-saving and time-saving, economic and reliable, and can shorten the screening period effectively, simplify the screening program, and save the screening material; and the sterile seedlings materials are obtained by seeds in a culture vessel directly, and enter the stage of root screening directly, which omits the trouble of cultivating adventitious buds as compared with the conventional screening test, so that manpower, material resources and time are saved. The tobacco sterile seedlings are adopted to screen the media, and the tobacco sterile seedlings screening can synchronize with the callus induction, so that the test schedule is speeded up. Besides, the induction effect of the method is consistent with that of a method for screening root media by using adventitious buds.

Description

Technical field: [0001] The invention relates to the technical field of plant tissue culture, in particular to a method for screening rooting medium by using sterile tobacco seedlings. Background technique: [0002] Plant tissue culture is not only a means of rapid plant reproduction, but also an ideal way for plant improvement, germplasm preservation and secondary biomass production. Common plant tissue culture needs to go through the process of callus-bud-sapling-rooting, such as: Chinese patent application 2008101469482 discloses the rapid propagation method of bald cypress in vitro, which directly selects seeds as explants, and then uses seeds to induce adventitious buds , and then induce rooting to cultivate strong seedlings, and directly select the medium when inducing rooting. Another example: Chinese patent application 2005100298416 discloses a kind of broad bean in vitro culture method, which selects broad bean seeds to cultivate aseptic seedlings, uses the seedlin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 张建慧殷英余祥文屈建康赵宇廖华李方楼徐伟
Owner 四川省烟草技术中心
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