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CN-121970682-A - Method for obtaining cyclamen regenerated plants by taking leaves as explants

CN121970682ACN 121970682 ACN121970682 ACN 121970682ACN-121970682-A

Abstract

The invention relates to the technical field of plant tissue culture, in particular to a method for obtaining a cyclamen regenerated plant by taking leaves as explants, which comprises the following steps of S1 taking cyclamen healthy leaves as explants, inoculating the cyclamen healthy leaves into an induction culture medium, inducing to form callus, S2 transferring the callus obtained in S1 into a proliferation culture medium for subculture, S3 inoculating the callus obtained in S2 into a differentiation culture medium, inducing to differentiate to form adventitious buds, and S4 transferring the adventitious buds obtained in S3 into a rooting culture medium, inducing to root, and obtaining the cyclamen regenerated plant. The invention takes cyclamen leaves as explants, induces callus to generate and induces differentiation to generate adventitious buds, can obtain cyclamen plants with stable inheritance, ensures the uniformity of seedlings while maintaining excellent properties, shortens the breeding period, and is suitable for commercial mass production.

Inventors

  • BAO YING
  • CHEN SHUYI
  • REN HONGYU
  • LI CHENXI
  • LI XUETING

Assignees

  • 唐山师范学院

Dates

Publication Date
20260505
Application Date
20260205

Claims (5)

  1. 1. A method for regenerating plants by taking leaves as explants, comprising the following steps: S1, taking healthy leaves of cyclamen as explants, inoculating the explants into an induction culture medium, and inducing to form callus; S2, transferring the callus obtained in the S1 into a proliferation culture medium containing 2, 4-dichlorophenoxyacetic acid with the concentration range of 0.1-1.5 mg/L for secondary proliferation culture; s3, inoculating the callus obtained in the step S2 into a differentiation medium, and inducing differentiation to form adventitious buds; S4, transferring the adventitious buds obtained in the step S3 into a rooting culture medium, and inducing rooting to obtain the regenerated plants of the cyclamen.
  2. 2. The method for obtaining a plant regenerated from cyclamen using leaf as explant according to claim 1, wherein the composition of the induction medium in S1 is 1/2MS medium, naphthalene acetic acid 0.15 mg/L, kinetin 0.2.2 mg/L, 6-benzylaminopurine 2.0 mg/L, mannitol 20 g/L, glucose 20 g/L, agar 7.5 g/L.
  3. 3. The method for regenerating plants by taking leaves as explants according to claim 1, wherein in S2, the proliferation medium has the composition of MS medium, 2, 4-dichlorophenoxyacetic acid 0.5 mg/L, kinetin 0.2.2 mg/L, 6-benzylaminopurine 1.0 mg/L, mannitol 20 g/L, glucose 20 g/L, agar 7.5 g/L.
  4. 4. The method for regenerating plants by taking leaves as explants according to claim 1, wherein in S3, the differentiation medium comprises MS medium, naphthalene acetic acid 0.5 mg/L, kinetin 0.2 mg/L, 6-benzylaminopurine 3.0 mg/L, mannitol 20g/L, glucose 20g/L, and agar 7.5 g/L.
  5. 5. The method for obtaining a regenerated plant of cyclamen using leaf as explant as claimed in claim 1, wherein the rooting medium in S4 is composed of 1/2MS medium, indole-3-butyric acid 0.5 mg/L, naphthylacetic acid 0.01 mg/L, sucrose 30 g/L, agar 7.5 g/L.

Description

Method for obtaining cyclamen regenerated plants by taking leaves as explants Technical Field The invention relates to the technical field of plant tissue culture, in particular to a method for obtaining a cyclamen regenerated plant by taking leaves as explants. Background Cyclamen (Cyclamen persicum) is a perennial bulb flower of the genus Cyclamen of the family primaceae. The Chinese cyclamen are more in variety, special in flower shape, gorgeous in color and long in flowering period, can reach five months, and the flowering period is just longer than the traditional festival such as the primordial denier, the spring festival and the like, so that the Chinese cyclamen are important potted flowers and flowers in the year night, and are popular with people as corm flowers with great ornamental value. At present, the seedling production of cyclamen mainly depends on seed reproduction. However, the mode has obvious limitations that on one hand, long-term selfing or inbreeding is easy to cause variety degeneration, and on the other hand, the newly bred or introduced fine variety is often subjected to serious segregation of characters in the sexual propagation process, so that the genetic stability and the phenotype consistency are difficult to maintain. In addition, part of the heavy-valve or sterile varieties cannot be propagated through seeds, so that popularization and application of the heavy-valve or sterile varieties are further limited. At present, the F1 generation seeds of the high-end cyclamen in China still depend on import, have high cost, and greatly restrict the development of cyclamen in China and the improvement of economic benefits. Plant tissue culture techniques provide an effective approach to solving the above-mentioned problems. By establishing a high-efficiency in-vitro regeneration system, the totipotency of plant cells can be utilized to induce the explant to form a complete plant through a callus or direct organogenesis way under the aseptic condition. The technology not only can realize rapid asexual propagation of excellent germplasm and ensure genetic consistency, but also can obviously shorten the breeding period. More importantly, the obtained callus or regeneration system can be used as an ideal receptor system, and lays a key technical foundation for genetic transformation, gene function research and molecular design breeding of cyclamen. However, the existing researches show that the in vitro regeneration process of cyclamen is influenced by various factors such as genotype, explant type and culture conditions, the regeneration capability difference among different varieties is obvious, and a regeneration method with strong universality and stable efficiency is not formed yet. Although regeneration attempts have been made using cotyledons, hypocotyls, flower holders, leaves, etc. as explants, there are general problems such as low callus induction rate, difficulty in differentiation, long regeneration cycle, and poor genetic stability. In particular, the regeneration system taking the leaves as the explants has larger application potential due to simple and convenient operation and sustainable material acquisition, but the efficient and stable technical scheme still lacks optimization and verification of the system. Therefore, there is a need to establish a tissue culture regeneration method which uses cyclamen leaves as explants, is simple and convenient to operate, has high regeneration efficiency and is genetically stable, so as to meet the actual demands of rapid propagation of good varieties and modern biological breeding. Disclosure of Invention The invention aims to provide a method for obtaining a cyclamen regenerated plant by taking leaves as explants so as to solve the problems in the prior art. In order to achieve the aim, the invention provides the following technical scheme that the method for obtaining the cyclamen to regenerate plants by taking leaves as explants comprises the following steps: S1, taking healthy leaves of cyclamen as explants, inoculating the explants into an induction culture medium, and inducing to form callus; S2, transferring the callus obtained in the S1 into a proliferation culture medium containing 2, 4-dichlorophenoxyacetic acid with the concentration range of 0.1-1.5 mg/L for secondary proliferation culture; s3, inoculating the callus obtained in the step S2 into a differentiation medium, and inducing differentiation to form adventitious buds; S4, transferring the adventitious buds obtained in the step S3 into a rooting culture medium, and inducing rooting to obtain the regenerated plants of the cyclamen. In a preferred embodiment, S1, the induction medium is composed of 1/2MS medium, naphthalene acetic acid 0.15 mg/L, kinetin 0.2.2. 0.2 mg/L, 6-benzylaminopurine 2.0 mg/L, mannitol 20 g/L, glucose 20 g/L, agar 7.5 g/L. In a preferred embodiment, S2, the proliferation medium is composed of MS medium, 0.5 mg/L of 2, 4-dichlorophenoxyaceti