One-step plantlet formation medium and method for isolated culture of lamiophlomis rotata
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A technology of in vitro culture and culture method, applied in horticultural methods, botanical equipment and methods, plant regeneration, etc., to achieve the effects of simple culture procedure, easy operation and short regeneration cycle
Inactive Publication Date: 2013-12-25
SICHUAN AGRI UNIV
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But, through the research to medium composition, adopt the technology of one-time tissue culture into seedling, also do not see the report
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Embodiment 2
[0038] In order to avoid the error of the experimental results caused by human factors, the applicant asked different researchers to inoculate 6 per bottle under the same conditions, and inoculate 8 bottles in total, using the unique one-step seedling growth medium identical to the embodiment, The same experimental operation as in Example 1 was carried out, and the experimental results are shown in Table 5 and Table 6.
[0039] Table 5 Effects of different hormone combinations on leaf callus induction
[0040]
[0041]
[0042] Table 6 Effects of different hormone combinations on the differentiation of adventitious buds from leaves
[0043]
[0044] Table 7 Survival rate of test-tube plantlets hardened by different hormone combinations
[0045]
[0046]
[0047]Analysis of the experimental data obtained in Table 5 shows that, except for the control group, the one-step seedling growth medium of each group can better induce the leaves of the unique herb, and its ...
Embodiment 3
[0053] A method for one-step seedling growth by in vitro culture of test-tube plantlet leaves comprising the following steps:
[0054] (1) Prepare the unique one-step seedling growth medium: add 2.0mg of 6-benzylaminopurine and 1.0mg of naphthaleneacetic acid to 1LMS medium, sterilize and set aside;
[0055] (2) Select the unique aseptic test-tube plantlets from the same source, and under aseptic conditions, cut out the fully expanded leaves at the top of about 0.5cm×0.5cm, and stick the back of the leaves on the medium described in step 1;
[0056] (3) Place the post-inoculation culture medium obtained in step (2) in an incubator at 20°C, and culture in complete darkness. After 7-10 days, carry out 16h / d light culture, with a light intensity of 1200Lx;
[0057] (4) After 55 days of light culture in the primary culture medium, regenerated plants of Duyiwei were obtained.
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Abstract
The invention discloses a one-step plantlet formation medium and method for the isolated culture of lamiophlomis rotata. By adopting the one-step plantlet formation medium, a lamiophlomis rotata test-tube plantlet leaf forms a plantlet in one step through isolated culture. The one-step plantlet formation culture medium is characterized in that a MS (Murashige and Skoog) medium is used as a basic medium, and 6-benzylaminopurine (6-BA), naphthylacetic acid (NAA) and 2,4-dichlorphenoxyacetic acid (2,4-D) which are different in concentrations and combinations are added to the base medium. According to the one-step plantlet formation method, the test-tube plantlet leaf serving as an explant is inoculated, then is induced by a callus and is directly differentiated into an adventitious bud and an adventitious root, so that the lamiophlomis rotata regenerated plantlet is obtained by one step. By adopting the one-step plantlet formation method, the callus obtained by induction in an initial medium does not need to be transferred into a differential medium for regenerating the plantlet. Compared with an isolated culture multi-step regeneration plantlet formation method, the one-step plantlet formation method has the advantages that the steps are simplified, the culture period is short, the repeatability is good, and the planting percent is high.
Description
technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a culture medium for one-step growth of seedlings in vitro, and a method for using the culture medium to make leaves of test-tube plantlets of only one step into seedlings. Correspondingly, the present invention also provides a A method for optimizing the medium. Background technique [0002] Lamiophlomis rotata (Benth.) Kudo belongs to Lamiophlomis rotata (Benth.) Kudo, a plant of the genus Lamiophlomis, and there is only one species of Lamiophlomis in this genus. It is mainly produced in Tibet, Qinghai, Gansu, western Sichuan and northwestern Yunnan; it grows in strongly weathered gravel beaches or stony alpine meadows and river beaches on plateaus or high mountains, at an altitude of 2700-4500m. It has the characteristics of typical alpine plants, such as dwarf plants, short stems, well-developed root system, large leaves, and spread out close to the ground. These ch...
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