Search

CN-122004126-A - Method for improving hardening-seedling transplanting survival rate of camellia tissue culture seedlings

CN122004126ACN 122004126 ACN122004126 ACN 122004126ACN-122004126-A

Abstract

The invention relates to a method for improving the survival rate of hardening-seedling transplanting of camellia tissue culture seedlings, which comprises the steps of sterilizing the tissue culture seedlings through ferrous sulfate disinfectant in the hardening-seedling process and then transplanting the tissue culture seedlings. The invention successfully solves the problem of huge economic loss caused by discarding the polluted or extremely precious camellia tissue culture seedlings, can realize the normal growth of the tissue culture seedlings under the condition of colony pollution, and greatly improves the survival rate of the tissue culture seedlings after hardening and transplanting.

Inventors

  • LI LINGLI
  • GUO TAO

Assignees

  • 重庆市风景园林科学研究院

Dates

Publication Date
20260512
Application Date
20241111

Claims (10)

  1. 1. A method for improving the survival rate of hardening-seedling transplanting of camellia tissue culture seedlings is characterized in that ferrous sulfate disinfectant is used for disinfecting the tissue culture seedlings in the hardening-seedling process and then transplanting is carried out.
  2. 2. The method for improving the survival rate of hardening-seedling transplanting of camellia tissue culture seedlings according to claim 1, wherein the ferrous sulfate disinfectant is FeSO 4 ·7H 2 O with the concentration of 20-50 mg/L.
  3. 3. The method for improving the survival rate of hardening-seedling transplanting of camellia tissue culture seedlings according to claim 2, wherein the ferrous sulfate disinfectant is FeSO 4 ·7H 2 O with the concentration of 35 mg/L.
  4. 4. A method of increasing the survival rate of hardening-seedling transplants in camellia tissue culture as claimed in claim 3, wherein said ferrous sulfate disinfectant is used for disinfection after bacterial or fungal contamination of said tissue culture seedling is observed.
  5. 5. A method for increasing the survival rate of hardening-seedling transplanting of camellia tissue culture seedlings as described in claim 4, wherein the amount of ferrous sulfate disinfectant used for the bacterially contaminated tissue culture seedlings is 5-10 mL times per use and 1-2 times per use.
  6. 6. A method for increasing the survival rate of hardening-seedling transplanting of camellia tissue culture seedlings as claimed in claim 4, wherein the amount of ferrous sulfate disinfectant used for the fungus-contaminated tissue culture seedlings is 5-10 mL for the first time, followed by 1-5 mL for each time until the contamination is eliminated.
  7. 7. A method for increasing the survival rate of hardening-seedling transplanting of camellia tissue culture seedlings as claimed in any one of claims 1-6, further comprising transplanting the tissue culture seedlings which normally grow after the disinfection after the soaking and cleaning of the tissue culture seedlings by ferrous sulfate disinfectant with the same concentration.
  8. 8. The method for improving the hardening-seedling transplanting survival rate of the camellia tissue culture seedlings according to claim 7, wherein the tissue culture seedlings after transplanting are cultured under the conditions that the culture medium comprises the main components of V turfy soil, V perlite which is 1:1 or 1:2, and auxin which is 0.5-1.0 mg/L, the pH value of which is 5.9-7.0, the air humidity of which is not lower than 90%, the illumination time of which is 16 h.d -1 , the dark culture of which is 8 h.d -1 , the illumination intensity of which is 50 mu mol.m －２ ·s －1 , and the temperature of which is 25+/-2℃.
  9. 9. A method for increasing the survival rate of hardening-seedling transplants of camellia tissue culture seedlings as claimed in claim 8, further comprising continuing to examine the tissue culture seedlings cultured after transplants for bacterial or fungal contamination, once every 2 days for 1 month after transplants, at least once a week for half a year after that, and once contamination is found, re-sterilizing under the above-mentioned sterilization conditions.
  10. 10. The application of ferrous sulfate in improving the survival rate of hardening-seedling transplanting of camellia tissue culture seedlings.

Description

Method for improving hardening-seedling transplanting survival rate of camellia tissue culture seedlings Technical Field The invention relates to the field of plants, in particular to a plant regeneration method by a tissue culture technology, and in particular relates to a method for improving the hardening-seedling transplanting survival rate of camellia tissue culture seedlings. Background The camellia is one of ten traditional flowers in China, at present, the cultivation of new varieties of camellia adopts traditional hybridization breeding, and the propagation of specific varieties adopts cutting, grafting and other methods. In traditional crossbreeding, full mature hybrid seeds without diseases and insect pests are required to be selected for sowing and seedling raising, and immature seeds generated by factors such as climate change, parent setting capacity and the like cannot be sowed and raised, wherein a large number of new varieties are contained. This portion of the seed is often thrown away or necrotic by decay as a result of natural selection. In the early work of the group of the present inventors, for example, in the technology disclosed in chinese patent documents CN113303227a and CN115413578a, mass propagation of new germplasm seedlings was achieved by constructing cell embryo clones with immature seeds of camellia japonica as explants. The technology realizes that under the aseptic condition, aseptic seedlings are induced through embryo rescue and somatic embryogenesis ways, regenerated plants are formed after hardening and transplanting, thereby greatly reducing adverse effects of natural environment on seed development, avoiding germplasm resource loss caused by seeding and seedling failure, simultaneously realizing rapid breeding of new variety seedlings, gradually culturing active absorption capacity of aseptic seedling fleshy roots on nutrition and moisture through loosening porous matrix hardening seedlings under the aseptic condition, and solving the problems of difficult survival and the like of hardening seedling transplanting of camellia tissue culture seedlings. However, in the process of germination of somatic embryos and growth of strong seedlings of camellia japonica, the culture period is about 6 months long, which often results in contamination caused by bacteria and fungi of the tissue culture seedlings, the contaminated tissue culture Miao Chang is sterilized by autoclaving to prevent the spread of the contaminated tissue culture seedlings, and the contaminated tissue culture seedlings need to be discarded. However, for the aseptic seedlings of camellia embryo which grow slowly, especially for distant hybridization of new germplasm, the quantity of the aseptic seedlings becomes the largest loss when the aseptic seedlings of tissue culture with smaller quantity are discarded once polluted. Therefore, how to continue the normal growth of the tissue culture seedling in the bacteria environment and find a balance point for the symbiotic of bacteria or fungi and the tissue culture seedling become difficult problems. The invention aims to overcome the defects of the prior art, and aims to develop chemical sterilization of the polluted camellia tissue culture seedlings with complete roots and buds in a bacterial environment in a sterile culture process, so as to realize bacterial and fungal death, but not influence the growth of the tissue culture seedlings, and even promote the further growth of the tissue culture seedlings, thereby laying a good foundation for hardening seedling transplanting. Disclosure of Invention In order to solve the technical problems, the invention provides a method for improving the hardening-seedling transplanting survival rate of camellia tissue culture seedlings, wherein ferrous sulfate disinfectant is used for disinfecting the tissue culture seedlings in the hardening-seedling process and then transplanting the tissue culture seedlings. Preferably, the ferrous sulfate disinfectant is FeSO 4·7H2 O with the concentration of 20-50 mg/L. Most preferably, the ferrous sulfate disinfectant is FeSO 4·7H2 O at a concentration of 35 mg/L. Further preferably, the ferrous sulfate disinfectant is used for disinfection after bacterial or fungal contamination of the tissue culture seedling is observed. More preferably, the usage amount of the ferrous sulfate disinfectant used for the bacterial-polluted tissue culture seedlings is 5-10mL each time, and the usage amount is 1-2 times. More preferably, the usage amount of ferrous sulfate disinfectant used for the fungus-polluted tissue culture seedlings is 5-10mL for the first time, and 1-5mL for each time is used until pollution is eliminated. And preferably, the method further comprises the step of transplanting the tissue culture seedlings which grow normally after the disinfection after the soaking and cleaning of the tissue culture seedlings with ferrous sulfate disinfectants with the same concentration. Furthe