Growing Plumeria from Seeds This is a colorless, sharp-smelling chemical which looks a bit like water. It is commonly used for household uses such as disinfectant and in cleaning products. It is Development and Standardization of Rapid and Efficient Seed Germination Protocol for Cannabis sativa Cannabis seed germination is an important process for growers and researchers alike. Many THE BASICS Like almost everything else about growing excellent Cannabis, germinating your seeds successfully is pretty simple. We’ve used this very basic, inexpensive method for many years and have shared it among thousands of growers worldwide who have all had excellent results with no issues. First make a mix in the
Growing Plumeria from Seeds
This is a colorless, sharp-smelling chemical which looks a bit like water. It is commonly used for household uses such as disinfectant and in cleaning products. It is rare to see pure hydrogen peroxide as it is most commonly used as a watered down solution.
It comes mixed with water which is typically 6% HP and the rest ordinary H20.
You may have heard of it as a hair bleach and peroxide is what is used by hairdressers to get a bright blonde hair color. It is used for medical problems like disinfecting small cuts or wounds and even treating boils or acne. It has a myriad of uses in household cleaning and it can even be used to kill mites and remove algae and scum from your home aquarium.
How Does It Work?
It is a very similar chemical composition to water and if it was in a sealed bottle you may even mistake it for water. And its effect on plants is actually quite positive.
But when it comes to gardening, this solution on plants does so much more than just simple water. Even though this is a chemical compound, it is actually found within rain naturally. Have you ever noticed how your plants seem to react better to rainwater rather than some from the tap? By soaking your plants in hydrogen peroxide solution, it actually replicates this natural substance found in raindrops and your plants drink it up, thinking that it’s ordinary rain!
How To Use Hydrogen Peroxide
Using hydrogen peroxide for gardening is pretty simple, but getting the mixture right is key. You want to soak your plants, around the roots, and get rid of fungus, spores and built up mold.
A general rule of thumb is to mix about one cup of HP solution with 32 cups of water and head out into the garden in the evening. Pour the solution into your pots or flower beds and leave it to soak overnight. Make sure you concentrate on the roots of your plant and avoid spraying the solution onto flowers or leaves.
You can also use a stronger solution to help seeds. One ounce of it as a solution in two cups of water will give you a great solution for seeds. This helps them stay healthy and grow faster.
1. Soil Aeration and Treatment of Root Rot
Good soil aeration is really important for any garden plants. Giving your soil enough space for air and nutrients to get through to the roots of your plants is the best way to see your plants flourish. If your soil is too compact, oxygen and nutrients won’t get through and your plants could die. A good way to see if your soil is too compact is to look at the roots of plants you have growing currently. If you pull out a plant and the roots look all squished together and tangled up then it means the soil doesn’t have enough aeration and your plant is struggling. You will also see more toxins and disease in your soil if it is poorly aerated.
Root rot is another problem that can crop up if your soil is too compact. Typically seen in plants which have been overwatered, it is the most common cause of decay around roots of plants and shrubs. Known as Phytophthora root rot, it doesn’t just affect plants in containers as bedding plants and bulbs can also suffer from this.
The difficult thing with root rot is that it can sit for years in your soil even if it hasn’t had any plants put in. It is difficult to treat because of this and is exacerbated when the ground is waterlogged or very compact. The most common time gardeners will see root rot is in potato and tomato gardens.
The signs you have root rot in a plant can be difficult to spot as it doesn’t appear above ground until it has well and truly taken over your plant. You will see yellow leaves and some branches dying off completely. If you dig around the roots of the plant you will see the roots are not formed well enough.
The good news is it can be treated with hydrogen peroxide. So, if you discover root rot or believe your soil is badly aerated, here is what you can do. It can and will kill off bacteria and fungus.
Use a weak solution (around 3%) and mix it roughly one part chemical to two parts water. Carefully pour it around the roots and the base of your plant to kill off the bacteria.
Hydrogen peroxide also helps aerate your soil which should help to prevent future cases of root rot. When it is absorbed into the soil, the hydrogen peroxide breaks down and releases oxygen. These high oxygen levels will make sure your roots are healthy and strong. A healthy root system should be long and untangled with fuzzy white growth on the main root which is used for soaking up water and nutrients.
2. Disinfect Growing Medium
There are hundreds of different types of growing media available. Put simply, a growing medium is a solid or liquid compound which is designed to help plants grow. This can be anything from a compost mix to a peat mix. Organic potting compost will have organic matter in it which can include bone meal, worm castings and even bat droppings! While this is great and packed full of nutrients for your garden and containers, you do run the risk of introducing bacteria and fungus into your garden through using this type of potting mix.
An easy way to stop disease infiltrating your garden this way is by disinfecting your chosen growing medium before you use it with a hydrogen peroxide solution.
Just put your chosen soil or potting mix into a clean, disinfected container and then sprinkle over some hydrogen peroxide mixture. The same sort of mix as you would use to pour over a plant suffering root rot. Allow this to settle in over a week or so, watering two or three times and then it should be ready for use in your garden or containers.
3. Sanitize Seeds
Everyone knows that seeds do best if they are soaked in water first, but did you know that using hydrogen peroxide in your water solution will help them germinate faster? Hydrogen peroxide will also kill off any bacteria that your seeds may have picked up. So if you’re buying them from another gardener you can make sure you get the plants you want without bringing in any unwanted diseases into your garden. Use a 3% hydrogen peroxide solution and pour it into a small, watertight container. Leave the seeds for around four hours and this will kill off bacteria without affecting the seeds.
You should then rinse the seeds thoroughly in clean, fresh water and then leave them to dry on a kitchen towel or a clean tea towel.
4. Accelerate Germination
Soaking seeds in hydrogen peroxide can also accelerate germination and even just a minute or two in a weak solution can speed up how fast your seeds will sprout. Put the seeds in a sieve or if you can a mesh bag works best. Something that will allow water through while keeping the seeds in place. Unlike bleach, hydrogen peroxide takes off any bacteria without damaging the seed itself. It should be a solution of one teaspoon in about a cup of water for the seed soak. You can use a much lower concentration but the soaking process will take much longer. You should rinse your seeds with water and then begin the germinating process straight away. You can use a thin layer of wet kitchen towels or sprinkle some potting soil into a seedling tray.
These trays are often made of cardboard or other absorbent material which will keep the seeds moist until they need to sprout. Read your seed instructions carefully and keep them at the right temperature. Warm and moist is best and, generally, seeds thrive in a warmer climate so think about putting them in a seed propagator or greenhouse.
5. Boost Root Development
If your plants are looking a little limp and lifeless, then give them a boost with a dose of hydrogen peroxide. While you can definitely use it as a seed booster and deal with fungal growths, it is also great for helping plants at any stage of their life do better. The hydrogen peroxide solution can be sprayed or absorbed straight into the soil to help boost your plant’s roots.
Watering plants with hydrogen peroxide solution will introduce more oxygen into your soil. This boost of oxygen gives the roots more room to take in extra nutrients and water to feed and it will give it a kick start or help an ailing plant get back on track. Use about two teaspoons of 35% hydrogen peroxide to around one gallon of water and then use it on your garden every other time you go out to water your plants.
6. Fight Fungal Infestions
One of the best uses for hydrogen peroxide is to fight fungal growth or mold on plants. You might be concerned about using something that will kill mold and spores around your precious plants but you don’t need to worry. This chemical mixture is brilliant at killing off fungus while also being kind to your shrubs.
If you spot the tell-tale signs that a fungus has taken hold of your plants, then mix up some hydrogen peroxide solution and water your plant really heavily. And I mean really heavily. You want to almost flood your plant and keep watering until the liquid pours out of the pot or the flowerbed is waterlogged.
Don’t panic! This might sound like bad advice but using this much “clean” water will completely rid the area, and soil around your plant, of the bacteria-carrying dirt and flush it clean. Don’t let your plants sit in a tray of water, though. Water them in one place and then move the container to another. Leave the watering until the plant has very nearly completely dried out and you should see the fungus die off. So, remember; hydrogen peroxide for powdery mildew, residues, and mold is the most effective and safe.
Mixing charts for gardening with hydrogen peroxide
If you want to start gardening with hydrogen peroxide, you need to know how much peroxide to use. Here are charts to tell you how much!
To water or mist plants, to soak seeds, to add to water used to wash sprouts:
|WATER||ADD THIS AMOUNT OF 3% HYDROGEN PEROXIDE||ADD THIS AMOUNT OF 35% HYDROGEN PEROXIDE|
|1 cup||1 and 1/2 teaspoons||7 to 10 drops|
|1 quart||2 tablespoons||1/2 teaspoon|
|1 gallon||1/2 cup||2 teaspoons|
|5 gallons||2 and 1/2 cups||3 tablespoons plus 1 teaspoon|
|10 gallons||5 cups||6 tablespoons plus 2 teaspoons|
|20 gallons||10 cups||3/4 cup plus 1 tablespoon plus 1 teaspoon|
To spray on sick or plants with fungus:
|WATER||ADD THIS AMOUNT OF 3%
|–OR– ADD THIS AMOUNT OF 35%
|1 cup||1 tablespoon||1/4 teaspoon|
|1 pint||2 tablespoons||1/2 teaspoon|
|1 quart||1/4 cup||1 teaspoon|
|1 gallon||1 cup||1 tablespoon plus 1 teaspoon|
|5 gallons||5 cups||6 tablespoons plus 2 teaspoons|
|10 gallons||10 cups||3/4 cup plus 1 tablespoons plus 1 teaspoons|
|20 gallons||20 cups||1 and 1/2 cups plus 2 tablespoons plus 2 teaspoons|
Please be mindful to choose the correct column in the chart depending on whether you are using 3% hydrogen peroxide or 35% hydrogen peroxide!!
As you may notice, the amount of peroxide in the chart for sick and fungus infected plants is twice as much as in the first chart. I have heard of people using stronger solutions, but more is NOT always better. So be careful, and when in doubt, stay safe. You can always apply more another day. If you decide to use a bit more, please make it only a bit more, don’t get carried away. Gardening with hydrogen peroxide is great, but too much can harm your plants. 10% hydrogen peroxide is recommended as a weed killer — in other words it will kill your plants at that concentration.
Development and Standardization of Rapid and Efficient Seed Germination Protocol for Cannabis sativa
Cannabis seed germination is an important process for growers and researchers alike. Many biotechnological applications require a reliable sterile method for seed germination. This protocol outlines a seed germination procedure for Cannabis sativa using a hydrogen peroxide (H2O2) solution as liquid germination media. In this protocol, all three steps including seed sterilization, germination, and seedlings development were carried out in an H2O2 solution of different concentrations; 1% H2O2 solution showed the fastest and the most efficient germination. This protocol also exhibited high germination efficiency for very old cannabis seeds with lower viability. Overall, this protocol demonstrates superior germination compared to water control and reduces the risk of contamination, making it suitable for tissue culture and other sensitive applications.
Keywords: Cannabis sativa, Rapid germination, Hydrogen peroxide, Seed sterilization, Seedling development
Cannabis sativa, otherwise known as marijuana or hemp, is an annual primarily dioecious flowering plant in which male/female sex is determined by heteromorphic chromosomes (X and Y) ( Gaudet et al., 2020 ). Cannabis is grown for a variety of agricultural uses; nearly all parts of cannabis plant are used, seeds for food, stem for fiber, and flowers/leaves for medicine. Flowers produce a mix of cannabinoids and aromatic compounds valued for their therapeutic and recreational effects ( Chandra et al., 2017 ). Cannabis plants are propagated either clonally through cuttings or via seed germination. Seed germination is very important for researchers, breeders, and growers alike, especially since seeds from elite cultivars can be very expensive and valuable. Additionally, older seeds may have a reduced germination rate while bacterial and fungal contamination can compromise germination, especially when seeds are germinated for tissue culture propagation. To address these issues, we have developed a rapid, sterile, and efficient seed germination protocol using a 1% hydrogen peroxide (H2O2) solution. In this protocol, all three steps including seed sterilization, germination, and seedlings development were carried out in a 1% H2O2 solution. This presents a significant advantage over other sterilants, such as mercuric chloride or bleach, which require additional washing of seeds and a separate germination step on MS solid medium. Our protocol resulted in faster germination and increased seed germination percentage as compared to water control, with no bacterial or fungal contamination, making it suitable for tissue culture and other sensitive applications. In comparison to previous germination methods which take between 4-7 days for radicle appearance and 5-15 days for seedling development ( Wielgus et al., 2008 and references therein), our germination method resulted in radicle appearance in 1 day and allowed us to obtain cannabis seedlings in a very short period (3-7 days) with minimal efforts. This protocol is also very efficient for germination of very old cannabis seeds with lower viability.
Materials and Reagents
All seeds were harvested in our laboratory. Blueberry seeds were not older than 6 months, when employed in the experiments. Finola and X59 seeds were more than 5 years old.
Growth chamber (Sanyo MLR-350, catalog number: 859-600-06): 24 °C, 18 h light/6 h dark cycle, light intensity 200 μmol·m -2 ·sec -1
Seed germination assay
Soak seeds overnight in various concentrations of hydrogen peroxide solution (liquid germination media or germination solutions) as well as in sterile water control (H2O, 1% H2O2, 3% H2O2, 5% H2O2, or 10% H2O2) in 15 or 50 ml screw-cap (Falcon tube). Falcon tubes with submerged seeds in various germination solutions were kept in the dark at room temperature.
Next day, record the percentage of germinated seeds in germination solution (appearance of radicle is considered as germination event) and add fresh respective germination solution after removal of old solution simply by pouring out.
Keep seeds soaked in the same solution for 3 more days in the dark at room temperature and record the percentage of germinated seeds every day.
Thereafter, germinated seeds/seedlings were transferred with or without seed coats from H2O2 solution to MS medium plates to observe the growth of H2O2 solution-germinated seeds/seedlings on MS medium. To transfer, first germinated seeds/seedlings were poured together with H2O2 solution from the Falcon tube to the empty petri plate. Then seedlings were transferred to sterile paper by using forceps to remove excess H2O2 solution. Finally, the germinated seeds/seedlings were transferred to MS media plate by using forceps. The whole transfer process has been carried out in the laminar flow hood.
Parafilm sealed MS medium plates with germinated seeds/seedlings are then transferred to the growth chamber (24 °C, 18 h light/6 h dark cycle and light intensity 200 μmol·m -2 ·sec -1 ) for 3 days to observe the growth and survival of H2O2 solution germinated seeds/seedlings on MS medium.
The H2O2 solution-germinated seeds/seedlings growth was also observed in soil. Pro-Mix HP Mycorrhizae Growing Medium used for soil experiment. The cannabis seeds were soaked in the H2O2 solution (germination solutions) for four days and thereafter, germinated seeds/seedlings were transferred from H2O2 solution to soil pot (Pro-Mix HP Mycorrhizae Growing Medium) to observe the growth and survival of H2O2 solution germinated seeds/seedlings on soil. The soil pots were transferred to the growth chamber (24 °C, 18 h light/6 h dark cycle and light intensity 200 μmol·m -2 ·sec -1 ). The photographs were taken on day 12.
Mean seed germination percentage under various concentrations of H2O2 solution as well as water control were calculated in an excel sheet. Data were shown as mean ± SE.
In this study, we have described a rapid and efficient seed germination protocol for Cannabis sativa. The brief description of this protocol has been reported in Sorokin et al. (2020) . In the current study, we have standardized the optimum concentration of hydrogen peroxide (H2O2) solution media for efficient sterilization and rapid germination. We have tested various concentrations of H2O2 solution as well as sterile water control (H2O, 1% H2O2, 3% H2O2, 5% H2O2, or 10% H2O2) for sterilization and germination efficiency. All three steps of germination (seed sterilization, germination, and seedlings development) were carried out in various concentrations of H2O2 solution and seeds were kept in liquid media for four days. Hydrogen peroxide presents several significant advantages over mercuric chloride or bleach sterilants, which require additional seed washing, and separate germination/seedling development step in Murashige and Skoog (MS) agar medium ( Sorokin et al., 2020 ). The 1% H2O2 solution showed rapid and higher germination than higher H2O2 concentrations solution and water control at day 1 ( Figure 1 ). On day 1, 1% H2O2 solution exhibited 82.5% germination as compared to 22.5% germination for 3% H2O2 group, 17.5% germination for 5% H2O2 group and 47.5% germination in water control group ( Figure 1B ). Interestingly, 10% H2O2 did not show any germination on day 1 due to its toxic effect ( Figure 1 ). In 1% H2O2 solution, radicle appearance (germination) occurred within 24 h and seedling development (two fully developed cotyledons and two immature true leaves stage) occurred in 72-96 h ( Figure 1A ). In comparison to previous germination methods which take between 4-7 days for radicle appearance and 5-15 days for seedling development ( Wielgus et al., 2008 and references therein), our germination method resulted in radicle appearance in 1 day and allowed us to obtain cannabis seedlings in a very short period (3-7 days) with minimal efforts ( Figures 1 -2). Considering the possible toxic effect of H2O2 (since germinated seeds/seedlings stayed continuously in H2O2 solution for 4 days), we have checked further survival of germinated seeds/seedlings on MS media and soil ( Figures 2 -3). On MS media, 1% H2O2 solution seedlings survived better than other treatments ( Figure 2 ). The water germinated seeds exhibited contamination and did not survive on MS media ( Figure 2 ). Similarly, due to the toxic effect of higher concentration of H2O2, the 10% H2O2 germinated seeds did not survive on MS media ( Figure 2 ). The 1% H2O2 solution seedlings also survived well on soil ( Figure 3 ). Apart from this, we have also tested our method for more than 5-years old cannabis seeds with lower viability, which demonstrated that 1% H2O2 solution medium exhibited a very high germination percentage (~50%) as compared to water control (~10%) ( Figure 4 ). In conclusion, we have developed a rapid and efficient method for C. sativa seed germination under sterile conditions for tissue culture and other sensitive applications.
Germination of 6-month-old seeds of Blueberry variety in various concentrations of hydrogen peroxide solution and water control.
A. Representative photographs of germinated seeds/seedlings in the H2O2 solution of various concentrations or water control on day 1 to day 4. B. Comparison of germination percentage between the various concentrations of H2O2 solution or water control. Data are shown as mean ± SE (n = 4). In each replicate, 30 seeds were used.
Representative photographs of growth and survival of H2O2 solutions germinated seeds/seedlings of Blueberry variety on MS media.
The Blueberry variety seeds were soaked in the H2O2 solution (germination solutions) for four days and thereafter, germinated seeds/seedlings were transferred from H2O2 solution to MS medium plates to observe the growth and survival of H2O2 solution germinated seeds/seedlings on MS medium. The photographs were taken at day 0 (just after transfer to MS medium plates), day 1 (after 24 h of the transfer to MS medium plates), and day 3 (after 72 h of the transfer to MS medium plates) on MS media.
Representative photograph of Blueberry variety young plantlet growing in soil (Pro-Mix HP Mycorrhizae Growing Medium).
The Blueberry variety seeds were soaked in the H2O2 solution (germination solutions) for four days and thereafter, germinated seeds/seedlings were transferred from H2O2 solution to soil pot (Pro-Mix HP Mycorrhizae Growing Medium) to observe the growth and survival of H2O2 solution germinated seeds/seedlings on soil. The photographs were taken on day 12.
Germination of 5-years old seeds of Finola and X59 varieties in 1% hydrogen peroxide solution and water control.
Comparison of germination percentage between 1% H2O2 solution media and water control. Data are shown as mean ± SE (n = 5). In each replicate, around 30 seeds were used.
4.43 g Murashige & Skoog Basal Medium with Vitamins
Adjust pH to 5.7 with KOH and sterilize by autoclaving at 121 °C for 40 min. 25 ml of MS media on each Petri plate.
This protocol is derived from Sorokin et al. (2020). We thank the Natural Sciences and Engineering Research Council of Canada (NSERC) and MITACS for funding our work.
The authors declare that they have no competing interests.
Readers should cite both the Bio-protocol article and the original research article where this protocol was used.
1. Chandra S., Lata H. and ElSohly M. A.(2017). Cannabis sativa L.-botany and biotechnology. Chandra, S., Lata, H. and ElSohly, M. A.(Eds.). Springer International Publishing: Cham, Switzerland. ISBN: 9783319545639. [Google Scholar]
2. Gaudet D., Yadav N. S., Sorokin A., Bilichak A. and Kovalchuk I.(2020). Development and optimization of a germination assay and long-term storage for Cannabis sativa pollen . Plants 9 : 665. [PMC free article] [PubMed] [Google Scholar]
3. Sorokin A., Yadav N. S., Gaudet D. and Kovalchuk I.(2020). Transient expression of the β-glucuronidase gene in Cannabis sativa varieties . Plant Signal Behav 15 ( 8 ): 1780037. [PMC free article] [PubMed] [Google Scholar]
4. Wielgus K., Luwanska A., Lassocinski W. and Kaczmarek Z.(2008). Estimation of Cannabis sativa L. tissue culture conditions essential for callus induction and plant regeneration . J Nat Fibers 5 : 199-207. [Google Scholar]
Like almost everything else about growing excellent Cannabis, germinating your seeds successfully is pretty simple. We’ve used this very basic, inexpensive method for many years and have shared it among thousands of growers worldwide who have all had excellent results with no issues.
First make a mix in the ratio of 1/3 3% hydrogen peroxide to 2/3 distilled water.
It’s important to use only distilled water – some of the common chemicals in tap water can stunt, warp or kill your plant even well past seedling stage when you’ve already spent weeks tending it.
Then soak your seeds in the distilled water/hydrogen peroxide mixture overnight. If you’re growing multiple varieties, be sure to soak them separately. (Don’t laugh. It’s happened.)
Next, pour some of the soak water onto really absorbent paper towels, then wring or squeeze them out lightly and lay them flat. We use 2-3 towels layered together to make a nice thick absorbent nest.
The best seeds often sink and compromised seeds sometimes float, but there are always exceptions to the rule.
Then sprinkle the soaked seeds, using a clean spoon or gloved fingers, onto the moist paper towels, not crowding the seeds.
Then fold the moistened towels over the seeds to make a flat little package.
If you’re sprouting more than 12 seeds at a time, make more than one package – don’t crowd them.
Then put the moist towel and seeds flat inside a closed, unzipped storage-size plastic baggie, laying it flat somewhere away from direct light at room temperature.
In two or three days the seeds will sprout – maybe not all at once, but that’s not a problem because within 24 hours of the first seed sprouting all the others will have sprouted too. Keep your inspection peeks short as not to dehydrate the paper towel.
When the seeds each have a 3/4” root and the halves of the shell are beginning to open noticeably, meaning the embryo leaves are swelling inside, they’re ready.
You then want to move each sprouted seed into a Jiffy Cube you’ve prepared by making a small hole using a pencil or chopstick. Again – be sure you used distilled water to hydrate your Jiffy Cubes and of course use it everywhere else during germination.
Replanted into Jiffys:
Using your fingers, pick up the seed very gently by its shell, and avoid touching the sprouting root as you transfer it into the jiffy cube.
Also remember what’s unfolding inside that little shell as you handle it, be super delicate and stay conscious of the life emerging inside.
Now place the sprouting seed root-down in the hole letting it settle in naturally with the top of the seed even with the top of the hole – never push it down.
If it doesn’t nest right in, lift it out and poke the hole a little deeper. The cells at the tip of that little root hold the most miraculous ecosystem of emerging life one can imagine and while they are incredibly tough in nature they are also vulnerable to our mis-handling.
Now all you have to do is let the new sprouts do their thing they do so well, with a little help from their friends. With a day or so they will raise up their first leaves. Keep them lightly misted if you’re in a dry environment but don’t over-do it.
In a couple of days when they’ve developed their first set of true leaves, not the embryo leaves, it’s time to put them into larger pots with living soil and a mild fertilizer and then let the plants fully establish themselves.
3 Days Later:
10 Days Later:
The same Skunk #1 x Haze seeds are available at:
Is using distilled water really important for germination?
Using distilled water is probably the most important part of the germination process because well water, tap water, bottled water and even rainwater contain traces of contaminants that can inhibit germination and later vegetative growth and flowering.
High chlorine in tap water, herbicides in well water, phthalates in bottled water, as well as many other kinds of common water contaminants can kill or damage germinating seeds and will definitely affect your plant’s health, yield and flower quality.
Can I remove these contaminants with a garden hose filter?
Yes most of them you can. There are a variety of relatively inexpensive filters that screw onto the end of the garden hose that you use to fill containers and water your plants. Some have replaceable screens, while others use granular activated charcoal.
There are also screw-on systems with carbon block filters that run $20-$30 and are very effective at reducing chlorine and chloramine, and at removing pesticides, heavy metals, and herbicides.
What’s some of the science behind using Hydrogen Peroxide for germination?
Here are two core articles from the PubMed database:
“Different Modes of Hydrogen Peroxide Action During Seed Germination”