Succulents, known for their fleshy leaves and drought-resistant characteristics, have gained popularity among plant enthusiasts. Tissue culture provides an efficient method for the propagation of succulents, eg. Haworthia cooperi Baker. Succulent tissue culture involves the propagation of these unique plants through the cultivation of their tissues in a controlled environment. This method offers a precise and efficient way to reproduce succulents while preserving their distinctive traits. Let's embark on a journey of discovery, unlocking the secrets that make tissue culture an indispensable tool for succulent enthusiasts.
1.Selection and Preparation of Healthy Stems:
Successful tissue culture starts with choosing robust and healthy specimens. Opt for mature plants with vibrant foliage and strong growth patterns.
Remove outer aging leaves. Rinse under tap water for 2 hours.
2.Sterilization:
Sterilization is a critical step in ensuring a contamination-free environment for tissue culture. Delicate succulents, like the renowned Haworthia cooperi Baker, demand meticulous attention to detail during the sterilization process. Disinfect with 75% alcohol for 10 seconds. Followed by 0.1% mercuric chloride (HgCl2) disinfection for 7 minutes. Rinse thoroughly with sterile water 5-6 times.
3.Creating the Ideal Growing Conditions:
Temperature, lighting, and moisture play pivotal roles in the success of succulent tissue culture. Maintain a temperature range of 25-28°C to promote optimal growth. Strategic lighting, avoiding prolonged exposure to high temperatures.
Light Intensity: 2000lx; Daily Light Exposure: 10 hours.
4.Cutting and Inoculation:
Cut sterilized material into segments with apical or axillary buds.
Inoculate segments onto induction medium (MS + 6-BA 3mg/L + NAA 0.2mg/L,3% sucrose, 0.6% agar, pH 5.8). After 18 days, induction rate can reach 80%.
5.Rapid Propagation:
After the induction period, transitioning to a proliferation medium fosters rapid multiplication. Transfer induced buds to proliferation medium (MS + 6-BA 1mg/L + NAA 0.2mg/L,3% sucrose, 0.6% agar, pH 5.8). After 20 days, proliferation multiplication can reach 5-7 times.
6.Rooting Culture:
Rooting culture marks a crucial stage in the tissue culture journey. Transfer the well-established shoots to a rooting medium, ensuring an ideal balance of growth regulators. Transfer buds (above 2cm) to rooting medium (1/2MS + 6-BA 0.1mg/L + NAA 2mg/L,3% sucrose, 0.6% agar, pH 5.8). After 15 days, rooting rate exceeds 75%, with each plant having 3-4 roots.
7.Acclimatization and Transplanting:
As roots develop, acclimatization becomes the final step before transplantation. After 4-5 days of hardening off the seedlings, remove roots from agar. Plant in a sterilized substrate (peat: coarse sand: perlite = 6:2:2).
Cover with plastic film for moisture retention. Once new roots emerge, transfer to a nursery bed, achieving a survival rate of about 87%.
Reference: Song, Y. (2014). Tissue Culture and Rapid Propagation Techniques of Ice Lantern Jade Dew. Modern Agricultural Science and Technology, 18, 164-166.