Clones, Mother plants, How to "Clean" cuttings!

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Cannabis sativa is a complex species with highly variable morphological features. The present chapter provides detailed descriptions of morphological and anatomical characters of various parts of C. sativa plant and illustrated with bright-field and scanning electron micrographs. Male and female flowers occur in separate plants. Three types of glandular trichomes namely, glandular stalked, glandular sessile and bulbous glandular trichomes are found. Of these, glandular stalked trichomes are restricted to the floral bracts in pistillate plants and anthers in staminate plants. The other two types of glandular trichomes are found in various parts including bracts, leaves, stems and petioles. Two types of non-glandular trichomes namely, cystolith trichomes and slender covering trichomes, are present. Cystolith trichomes are primarily found on the adaxial leaf surface while the covering trichomes are commonly present on the abaxial leaf surface, stems, petioles and tepals. Cystolith crystals of calcium carbonate and cluster crystals of calcium oxalate are observed in the leaves. Anatomical features of various parts of the plant are described and illustrated.

Morphology of C. sativa. a-c Twigs with female inflorescences; d A twig of a male plant; e Leaves showing variation in the number of lobes; f Seeds
…

Micro-morphology of different parts of C. sativa a, c, e, f Scanning Electron Microscopy (SEM); b, d Light microscopy (LM)]. a Female flowers; b Outer surface of a bract showing numerous glandular trichomes with yellow contents; c Portions of bract and young leaves with various types of trichomes; d A portion of bract; e Petiole surface covered with nonglandular and glandular trichomes; f Lower surface of leaf base and a portion of petiole showing branching of major veins to leaf lobes
…

Leaf micro-morphology of C. sativa [C and F-LM; all others SEM]. a Adaxial leaf surface; b, c Adaxial leaf epidermis; d, e Abaxial leaf surface; f, g Abaxial leaf epidermis showing stomata. Cc cystolith trichome, Cu cuticle striations, Gt-2 capitate sessile glandular trichome, Gt-3 bulbous glandular trichome, Ngt non-glandular trichome, St stomata
…

Anatomy of C. sativa [A and C-LM; all others SEM]. a, b Transection (TS) of leaf through midrib; c, d TS of leaf through lamina; e TS of stem, with a portion enlarged (f). Ch chlorenchyma, Co collenchyma, Ct cystolith trichome, Fu furrows, Gt-2 capitate sessile glandular trichome, Gt-3 bulbous glandular trichome, La lamina, Ld laticifer duct, Le lower epidermis, Mr midrib, Ngt non-glandular trichome, Pa palisade, Pf pericyclic fibers, Ph phloem, Pi pith, Ri ridges, Sp spongy tissue, Up upper epidermis, Xy xylem
…

Anatomy of C. sativa [E-LM; all others SEM, F-colorized SEM]. a TS of petiole; b TS of root; c, d TS of root; e, f cluster crystals of calcium oxalate in the leaf midrib. Br bract, Cc cluster crystals, Co cortex, Co cotyledon, Gr adaxial groove, Pe pericarp, Ph phloem, Vb vascular bundle, Xy xylem
…
Figures - uploaded by Vijayasankar Raman
Author content
Content may be subject to copyright.

Morpho-Anatomy of Marijuana (Cannabis sativa L.)​

• May 2017

DOI:10.1007/978-3-319-54564-6_5

 

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Callus of Cannabis sativa has been successfully induced from C. sativa explants and seedings. It seems that flowers are the best explant for callus induction and induction under light also give better results than induction in dark.

CALLUS INDUCTION AND PHYTOCHEMICAL ...​

Jurnal Universitas Gadjah Mada
https://journal.ugm.ac.id › ijc › article › download

Recalcitrance of Cannabis sativa to de novo regeneration​

National Institutes of Health (NIH) (.gov)
https://pmc.ncbi.nlm.nih.gov › articles › PMC8363012


by AS Monthony · 2021 · Cited by 48 — Callus formation was observed as soon as one week after induction in the earliest genotype, with all genotypes producing callus within two weeks ...

Cannabis sativa callus induction stages: (a) initiation of ...​

ResearchGate
https://www.researchgate.net › figure › Cannabis-sativa-...


Cannabis sativa callus induction stages: (a) initiation of callus, (b) callus formation 1.5 weeks after culture, (c) growth of the callus, and (d) cell death.

Regeneration from C. sativa leaf explants (replication study)

Hosted on the Open Science Framework

osf.io

 

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Tissue culture experimental system for flower induction in cannabis. a Stem segment explants, cut from indoor TA5 plants growing under 18/6 h light/dark photoperiod, were introduced into tissue culture (a white circle marks a node segment). b Stem section explants were surface sterilized in a 2% sodium hypochlorite solution. c The stems were separated into single-node segments, and five explants were cultured in each vessel. d Developed plants at two weeks under an 18/6 h light/dark photoperiod. e Cannabis flowers developed under a 12/12 h cycle to promote flowering. Images were taken three weeks into the flowering photoperiod. f A close-up of an in vitro cannabis flower (bar = 100µm). g and h Comparison between two photoperiod regimes. Node segments were introduced to tissue culture on the same date. g Vessels were immediately cultured under a flower-inductive photoperiod (12/12 h). h Vessels were cultured for two weeks under an 18/6 h cycle before switching to 12/12 h. Images were taken two weeks after introducing to tissue culture. i Average number of flowers per plant under two photoperiod regimes, with counts taken at specified times after introducing to tissue culture. The average flower number was calculated for five plants per vessel, presented as mean ± SE per plant for eight vessels for each treatment (n = 8); different letters represent a significant difference at a p < 0.05 using the Student's t-test
https://plantmethods.biomedcentral.com/articles/10.1186/s13007-024-01265-5/figures/1
Optimizing cannabis cultivation: an efficient in vitro system for flowering induction.
Plant Methods 20, 141 (2024). https://doi.org/10.1186/s13007-024-01265-5

 

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Table 1. Experimental conditions in the current study compared to Lata et al. (2010) [11].​

Experimental Condition​	Current Study​	Lata et al. (2010) [11]​	Comment​
Plant material and Preparation​
Genotype​	Clonal Genotypes: U22, U31, U37, U38, U42, U61, U82, U91 and GRC Heterogenous (seed derived) Plants: RTG​	Clonal Genotype: MX​	MX described as a high THC elite female clone [11,12]. See Fig 3. for the genotypes used in the current study.​
Explant Source​	Young, fully expanded in vitro leaves, found no lower than 3 nodes below the shoot apex.​	Young ex vitro leaves from plants grown in a controlled environment growth room.​	Criteria for selecting leaves (location, ages, etc.) N.S. in Lata et al. (2010) [11].​
Explant Type and Preparation:​	Young leaves, cut into squares using sterile disposable scalpels.​	Young leaves​	Explant preparation not specified in Lata et al. (2010). [11]​
Explant Size:​	100 mm2​	0.5–10.0 mm​	N/A​
Culture Conditions​
Light Type:​	LED​	Fluorescent​	Type of fluorescent not stated by Lata et al. (2010) [11]​
Photoperiod:​	16 h​	16 h​	N/A​
Light Intensity:​	48.74 ± 3.53 μmol s-1 m-2​	~ 52 μmol s-1 m-2​	N/A​
Light Spectrum:​	See S1 Fig.​	N.S.​	Light spectra were not provided by Lata et al. (2010) [11]​
Vessel Type:​	5.8 cm × 9.05 cm baby food jars with magenta B caps.​	4 cm × 9.5 cm baby food jars with magenta B caps.​	N/A​
Media Composition and Preparation​
Basal Salt:​	Murashige and Skoog’s medium (Product ID: M524; Phytotechnology Laboratories)​	Murashige and Skoog’s medium (supplier unknown)​	N/A​
Plant Growth Regulators​	0.5 μM NAA (Sigma Aldrich) 1.0 μM TDZ (Caisson Laboratories, Inc.)​	0.5 μM NAA 1.0 μM TDZ (Sigma Aldrich)​	N/A​
Gelling Agent​	0.8% (w/v) type E agar (Sigma Aldrich)​	0.8% (w/v) type E agar (Sigma Aldrich)​	N/A​
pH​	5.7​	5.7​	N/A​
Sterilization​	20 minutes at 121°C and 18 PSIG.​	Media was sterilized. (conditions N.S.)​	N/A​

Open in a new tab
N.S.- not specified; N/A- not applicable.
* Reported as standard error of the mean.


Young, fully expanded in vitro leaves, found no lower than 3 nodes below the shoot apex.

Light Type:	LED	Fluorescent	Type of fluorescent not stated by Lata et al. (2010) [11]
Photoperiod:	16 h	16 h	N/A
Light Intensity:	48.74 ± 3.53 μmol s-1 m-2	~ 52 μmol s-1 m-2	N/A

D) Green, nodular callus of C. sativa cv. U61, one month after culture LT-C medium.

 

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The extent of development of the central pith tissues in cannabis stem cuttings. The cuttings were obtained from stock plants at the same stage of growth and sequential stem pieces were taken (from top to bottom) and sectioned by hand and 3 mm thick slices were placed on the observation stage of a dissecting microscope and photographed at the same magnification. (a, b) Early stage of pith development with densely packed central parenchyma cells. (c, d) Loosely packed parenchyma cells in the pith region. (e, f) Development of hollow central pith where parenchyma cells are absent. (g, h) Complete development of central pith in stem cutting of cannabis strain 'Hash Plant'. Scale bars shown in a and b apply to all figures.​

 

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TOPIC 9.2: TRANSPORT IN THE PHLOEM OF PLANTS​

image from Dokimi Science

In the Transport in the Phloem unit we will learn that the phloem is the living tissue that transports glucose and other soluble compounds to parts of the plant where needed. This transport process is called translocation This unit will last 3 school days

Essential idea:

• Structure and function are correlated in the phloem of plants.

Nature of science:

• Developments in scientific research follow improvements in apparatus—experimental methods for measuring phloem transport rates using aphid stylets and radioactively-labelled carbon dioxide were only possible when radioisotopes became available. (1.8)
  • Outline how radioactive carbon isotopes are used to study translocation.

Understandings
9.2 U 1 Plants transport organic compounds from sources to sinks.

• Define translocation, phloem sap, source and sink.
• List example source and sink tissues.
• State that phloem transport is bidirectional.

Translocation is the movement of organic compounds (e.g. sugars, amino acids) from sources to sinks. Organic molecules such as sucrose and amino acids move from a source to a sink via phloem tubes in plants.

The phloem is composed of living tissue called sieve tube members (lack a nucleus) that are joined end to end to form a tube that conducts food materials throughout the plant. They are bordered by companion cells that carry out the cellular functions of a sieve-tube element. Phloem tubes can carry sugars and amino acids in a variety of directions; depending on where the source and the sinks are located (sometimes roots can be sources or sinks).

Sources produce sugars by photosynthesis in leaves or green stems or by hydrolysis of starch in storage vessels (germinating seeds or roots/tubers) and deliver these products via the phloem to the sink (roots, buds, stem, seeds, and fruits). At the source, sugar and other organic molecules are loaded into the sieve tube members thus increasing solute concentration within the sieve tube cells (decreases water potential).

Water from surrounding tissues, enters the sieve tube members by osmosis following a concentration gradient.. The water absorbed into the sieve tube creates hydrostatic pressure that forces the phloem sap to flow (bulk flow) towards the sink.

image from plant cell biology
9.2 U 2 Incompressibility of water allows transport along hydrostatic pressure gradients.

• Outline why pressure in the phloem increases due to the movement of water into the phloem.

At normal temperature, the compressibility of water is so low that it can be considered incompressible. Any change in pressure at one end of the transport vessel will result in an equal change in pressure at the other end of the transport vessel. This allows phloem sap to move up or down, with or against gravity, as it moves from source to sink.

Build-up of sucrose and solutes, causes water to enter the companion cells through osmosis. The incompressibility of water and ridged cell walls causes a build-up of pressure. Water flows from high pressure to low pressure
Sucrose is used as an energy source at the sink for growth or converted to starch for storage, reducing the amount of sucrose and thus reducing osmotic pressure. Water that carried the solute is drawn back into the xylem

9.2 U 3 Active transport is used to load organic compounds into phloem sieve tubes at the source

• State that sucrose is the most prevalent solute in phloem sap.
• Outline why sucrose is used for phloem transport, as opposed to glucose.
• Describe the active transport of sucrose into the phloem via a co-transport protein.

Organic compounds produced at the source are actively loaded into phloem sieve tubes by companion cells. Sugars are actively unloaded at the sink and water returns into the xylem tubes reinforcing the pressure gradient from the source to the sink. Water is recycled by the xylem, returning it from the sink back to the source.
At the source sugars are loaded through active transport in a process called phloem loading

In some plants sugars travel through cell walls from mesophyll cells to cell walls of companion cells and some sieve cells. Sugar is then actively transported into the phloem by a sucrose transport protein (Apoplast Pathway). Protons are pumped out of the companion cells from the tissues by active transport, using ATP as an energy source. This creates a proton gradient. H+ binds with sucrose and flows back into the companion cell-sieve tube complex through a co-transport protein, following its concentration gradient, and pulling sucrose molecule with it into the cell.

In other plants, sucrose can travel between cells through connections called plasmodesmata (symplast route). Once the sucrose reaches the companion cell, it is converted to an oligosaccharide, which maintains the sucrose concentration gradient

image from IB Study Buddy
9.2 U 4 High concentrations of solutes in the phloem at the source lead to water uptake by osmosis

• State that the phloem becomes hypertonic to xylem due to the active transport of sucrose into the phloem.
• State that water moves into the phloem by osmosis.

The active transport of solutes (such as sucrose) into the phloem by companion cells makes the sap solution hypertonic. Loading of sucrose into the STMs at the source. Reduces the water potential inside STMs, causing water to enter by osmosis

9 2 U.5 Raised hydrostatic pressure causes the contents of the phloem to flow towards sinks.

• State that water moves from area of higher pressure to area of lower pressure and that the movement of water also moves the solutes dissolved in it.

At the source, sugar and other organic molecules are loaded into the sieve tube members thus increasing solute concentration within the sieve tube cells (deceases water potential). Water from surrounding tissues, enters the sieve tube members by osmosis following a concentration gradient. The water absorbed into the sieve tube creates hydrostatic pressure that forces the phloem sap to flow (bulk flow) towards the sink

The Pressure Flow Hypothesis

• The proposes that water containing food molecules flows under pressure through the phloem.
• The pressure is created by the difference in water concentration of the solution in the phloem and the relatively pure water in the nearby xylem ducts.
• At their "source" — the leaves — sugars are pumped by active transport into the companion cells and sieve elements of the phloem.
• As sugars (and other products of photosynthesis) accumulate in the phloem, water enters by osmosis.In the figure, sugar molecules are represented in black, water molecules in red.)
• Turgor pressure builds up in the sieve elements (similar to the creation of root pressure).
• As the fluid is pushed down (and up) the phloem, sugars are removed by the cortex cells of both stem and root (the "sinks") and consumed or converted into starch.
• Starch is insoluble and exerts no osmotic effect.
• Therefore, the osmotic pressure of the contents of the phloem decreases.
• Finally, relatively pure water is left in the phloem, and this leaves by osmosis and/or is drawn back into nearby xylem vessels by the suction of transpiration-pull.

---

Phloem Loading Animation

Application
9.2 A 1 Structure–function relationships of phloem sieve tubes.

• State that the function of phloem includes loading of carbohydrates at a source, transport of carbohydrates through the plant, and unloading of carbohydrates at a sink.
• Outline the structure and function of sieve tube cells, with specific mention of the rigid cell wall and sieve plates.
• Outline the structure and function of companion cells, with specific mention of mitochondria and cell membrane infolding

Skill
9.2 S 1 Identification of xylem and phloem in microscope images of stem and root

• State two ways xylem cells can be identified in cross sections of stem and root.
• Identify xylem given microscope images of stem and root.
• Identify phloem within the vascular bundle of a stem and root.

9.2 S 2 Analysis of data from experiments measuring phloem transport rates using aphid stylets and radioactively-labelled carbon dioxide.

• State that aphids consume phloem sap as the main component of their diet.
• Outline how aphids have been used to measure the rate of flow and composition of phloem sap.

Radioactively labeled carbon-14 contained within CO2 can be fixed by plants by photosynthesis . The carbon in these sugars created by photosynthesis, is metabolized by the plant and will be found in different molecules within the plant. This carbon can be detected using film or radiation detectors. Therefore the movement, use, and formation of these molecules can be traced

Aphid uses stylet to tap into phloem. The aphid body cut from stylet after stylet inserted into phloem. The phloem sap continues flowing through stylet. Analyse sap from solutes/carbohydrates. Plants are grown in radioactive CO2 which becomes incorporated into carbohydrates produced by plant. Radioactive-labeled carbon can be detected in the phloem sap. Stylets placed at different parts of the plant can show rate of movement of phloem sap.

 

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DKW Medium with Vitamins​

Contains the macro-, micronutrients, and vitamins as described by Driver & Kuniyuki (1984) and McGranahan, et al (1987).

This medium was developed for the multiplication of shoots from nodal explants. The medium was supplemented with 4.5 μM BA and 5 nM IBA. Rooting the shoots was enhanced by dipping the basal ends of the shoots in 5 mM IBA prior to transferring to the greenhouse.

Polyacrylamide - Wikipedia

en.wikipedia.org

DKW Media: Definition, Use, and Applications​

Anjali Singh, MS
As a content and community manager, I leverage my expertise in plant biotechnology, passion for tissue culture, and writing skills to create compelling articles, simplifying intricate scientific concepts, and address your inquiries. As a dedicated science communicator, I strive to spark curiosity and foster a love for science in my audience.

Table of Contents

Introduction​

In vitro plants need nutrients to grow in a lab environment. These media are composed of micronutrients and macronutrients that promote the growth and development of plant tissues.

Here's a glimpse into five widely used culture media:

• Murashige and Skoog (MS) medium: This is the king of plant tissue culture media, invented in 1962. It's a versatile blend of inorganic salts, vitamins, and amino acids used for various applications like organ formation, callus culture, and micropropagation.
• Linsmaier and Skoog (LS) medium: Developed in 1965, LS medium is similar to MS but with a twist: it uses a modified vitamin mix with a higher concentration of thiamine. This tweak proved successful for tobacco cultures. LS serves similar purposes as MS media.
• Gamborg (B5) medium: Introduced in 1968, the B5 medium is ideal for callus and cell suspension cultures, particularly for soybeans. It has a distinct composition with higher nitrate and potassium, favoring root callus formation, compared to the lower ammonia content that supports cell growth.
• Nitsch and Nitsch (NN) medium: Developed specifically for in vitro anther culture of tobacco, NN medium (1969) boasts a high concentration of thiamine, biotin, and folic acid to promote anther callus formation.
• White's Medium: This pioneering medium (1963) was formulated for tomato root cultures. With lower salt concentration and higher magnesium sulfate compared to MS, White's medium is suitable for shoot and callus cultures, particularly for banana and carrot species.

What is DKW Media?​

Basal mediums are nutrient solutions used to grow plant cells, tissues, or organs in a controlled environment. DKW media is also known as Driver and Kuniyuki Walnut medium. The media was originally developed for walnut micropropagation using nodal explant.

DKW is different from other basal mediums because it has a higher concentration of salts, specifically chemical macronutrients and minerals.

The solidifying agent used in DKW is called Gelrite, and researchers believe it plays a vital role in making DKW more effective than other mediums.

Two reasons for the effectiveness of DKW media:

• It uses a polysaccharide produced by Sphingomonaselodea bacteria, which reduces the need for another common solidifying agent, agar .
• It may enhance the responsiveness of plant cells to cytokinins, a type of plant hormone that promotes cell division and shoot growth.

While DKW seems promising, more studies are needed on various plant species to confirm its effectiveness definitively.



We now have DKW Media at the PCT Store – Shop here!

Composition of DKW Media​

Given below is the list of each component of DKW media and their concentration:

Component	mg/L
Ammonium Nitrate	1416
Boric Acid	4.8
Calcium Chloride	149
Calcium Nitrate	1367.47
Copper Sulphate	0.25
EDTA Disodium Salt	45.4
Ferrous Sulphate	33.8
Glycine	2
Magnesium Sulphate	361.38
Manganese Sulphate	33.5
Myo-inositol	1000
Nicotinic Acid	1
Nickel Sulphate	<0.250
Potassium Dihydrogen Phosphate	265
Potassium Sulphate	1559
Sodium Molybdate	0.39
Thiamine Hydrochloride	2
Zinc Nitrate	17

Applications​

DKW media use in walnut micropropagation​

DKW has been proven superior to other mediums for increasing the number of shoots in Persian walnut (Juglans regia L.) and black walnut (Juglans nigra L.). This means that using DKW medium can result in more walnut shoots growing in a shorter period.

DKW use in Cannabis micropropagation (over MS media)​

While a well-established protocol using Murashige and Skoog (MS) basal medium with thidiazuron (TDZ) exists for cannabis shoot multiplication, researchers observed limitations in their lab. Cannabis grown on MS-T05 medium exhibited poor health, low multiplication rates, and inconsistency across cultivars.

This led them to explore alternative basal media. DKW medium emerged as a promising option during initial screening, producing visibly healthier explants compared to MS. Further studies are underway to confirm DKW's effectiveness across various cannabis genotypes and pinpoint the specific factors contributing to its success.

The different composition of DKW, particularly higher levels of sulfur, calcium, and copper compared to MS, might be responsible for this positive effect. Future research aims to optimize DKW further and refine other aspects of the propagation system for even better results.

DKW Media Use In Woody Plant Micropropagation​

DKW medium is a promising option for woody plant micropropagation due to its high sulfate content, which has been linked to increased shoot multiplication. Studies have shown that sulfate concentrations above 2.85 mM are crucial for enhanced shoot multiplication in woody fruit trees. This aligns with findings where high levels of magnesium, manganese, zinc, and copper sulfates improved shoot formation in red raspberries. Further research is needed to fully understand the mechanisms at play and optimize DKW for a wider range of woody plant species.

​

How To Use DKW Media In Your Tissue Culture Applications?​

Follow the given steps to prepare DKW media for your tissue culture applications:

1. Dissolve 5.36gms of the medium in 600ml of distilled or deionized water at room temperature (15-30°C).
2. Rinse media vial with small quantity of distilled water to remove traces of power.
3. Add the desired heat stable supplements before autoclaving.
4. Continue stirring until the powder has dissolved.Make up the final volume to 1000ml with distilled water.
5. Mix Gently, heat and rotate between intervals until the solution becomes clear. Do not boil, reheat and allow to cool below 50°C during dispensing. Dispense the medium into suitable containers, plug or cap, then autoclave at 15lbs (121°C for 15 minutes.
6. Cool the autoclaved culture vessels containing medium to 45-50°C and aseptically add desired sterile heat-labile substrate.
7. Mix well and aseptically dispense desired quantity of the media in sterile culture vessels.

NOTE: Some times media does not dissolve completely unless the pH is reduced. For these, lower the pH to about 3.0 to facilitate dissolution of media. The pH of medium is adjusted by using 1N HCL/ 1N NaOH/ 1NKOH.

 

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Perlite - Wikipedia

en.wikipedia.org

Polyacrylamide - Wikipedia

en.wikipedia.org

Composition of DKW Media​

Given below is the list of each component of DKW media and their concentration:

Component	mg/L
Ammonium Nitrate	1416
Boric Acid	4.8
Calcium Chloride	149
Calcium Nitrate	1367.47
Copper Sulphate	0.25
EDTA Disodium Salt	45.4
Ferrous Sulphate	33.8
Glycine	2
Magnesium Sulphate	361.38
Manganese Sulphate	33.5
Myo-inositol	1000
Nicotinic Acid	1
Nickel Sulphate	<0.250
Potassium Dihydrogen Phosphate	265
Potassium Sulphate	1559
Sodium Molybdate	0.39
Thiamine Hydrochloride	2
Zinc Nitrate	17

2% SUCROSE

 

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Transform yourself from an average Joe-Schmoe to a seasoned Grow-Pro with Pro-Tips!
Do you know what’s growing in your reservoir? When was your last res change? Do you even air-lift brah?
Here are some quick tips to keep your reservoir from quickly becoming a breeding ground for algae, bacteria and protozoa.

1. Always aerate your reservoir: especially if you plan on using it over a few days. We recommend an air-stone paired with a quality air pump. Remember that larger reservoirs will require correspondingly larger air pumps; make sure your pump is up to the job!
2. Every few weeks, drain your reservoir and clean and sterilize any pumps, air-stones and the inside of the reservoir. Look into home brewer supply stores; items such as PBW and Star San work very well.
3. Organic nutrients and supplements are great, just remember they must be made to order and usually do not do well if kept in a reservoir over long periods of time.
4. Make sure your reservoir is covered and as light tight as possible. This will ensure that algal growth is minimized.

Air-lift pumps are used in hydroponics to circulate and aerate water. They use compressed air to create an air-water mixture that rises through a pipe.

How it works

1. Air is injected into the bottom of a pipe that's partially submerged in liquid
2. The air mixes with the liquid, creating a less dense mixture
3. The less dense mixture rises through the pipe

Benefits

• Energy efficient: Airlift pumps can reduce energy consumption by 50–70%
  
  
• Low maintenance: Airlift pumps are simple to install and require little maintenance
  
  
• Resistant to clogging: Airlift pumps are less likely to get clogged than other types of pumps
  
  
• Easy to control: Airlift pumps have a simple flow rate control
  

Applications

• Hydroponics
  Airlift pumps can be used to circulate water and aerate plants in hydroponic systems

​

 

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Polyacrylamide (PAAm) is a polymer hydrogel that can be used to improve plant rooting and soil health. PAAm can be used in soil conditioners, in vitro plant rooting, and to study root growth.


Soil conditioners

• Soil tilth: PAAm can improve soil tilth, aeration, and porosity.
• Water run-off: PAAm can reduce water run-off.
• Plant vigor: PAAm can increase plant vigor, color, and appearance.
• Rooting depth: PAAm can increase rooting depth.
• Disease: PAAm can decrease diseases.
• Erosion: PAAm can reduce erosion.

In vitro plant rooting

• PAAG-amber: A PAAm hydrogel enriched with amber powder can be used to stimulate plant rooting.
  
  
• Seed viability: PAAm can be used to determine the viability of plant seeds.

Root growth

• Polyacrylamide gel crystals: PAAm gel crystals can be used to study root growth by planting seeds in a clear container filled with water gel.

DOPED WITH DKW 2% SUCROSE

 

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Evaluating Propagation Techniques for Cannabis sativa L. Cultivation: A Comparative Analysis of Soilless Methods and Aeroponic Parameters​

Plants 2024, 13(9), 1256; https://doi.org/10.3390/plants13091256

MICRO PROPAGATION OF APEX IS PREFERRED OVER "TEEN" SIZED CUTTINGS
WASHING IN BLEACH AND ALCOHOL TRANSFER TO STERILE ROOTING MEDIA
ROOTING GEL, ONCE CUTTING WOUND IS HEALED (7-14 DAYS)
AND AGGRESSIVE ROOT GROWTH IS ESTABLISHED (14-28 DAYS)
TRANSFER TO HARDENING OFF MEDIA (WITHIN 60 DAYS)

Chapter 9 - Role and Regulation of Plants Phenolics in Abiotic Stress Tolerance: An Overview​

https://doi.org/10.1016/B978-0-12-816451-8.00009-5

 

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Figure 2 Root-infecting pathogens on Cannabis sativa. (A) Symptoms of brown discoloration on the root system of indoor hydroponically grown plants. (B) Colonies of Fusarium oxysporum isolated from diseased roots in (A) growing on potato dextrose agar. (C) Colony of Pythium catenulatum isolated from diseased roots growing on potato dextrose agar. (D) Symptoms of natural crown infection on a field-grown cannabis plant caused by a combination of F. oxysporum, Fusarium brachygibbosum, and Pythium aphanidermatum. (E) The crown area of the infected plant shown in (D) is sunken, and there is visible mycelial growth on the surface. (F) Colony of Fusarium brachygibbosum isolated from diseased roots growing on potato dextrose agar. (G) Symptoms of plant collapse as a result of infection by P. aphanidermatum under a greenhouse environment. (H) Comparison of a noninoculated plant (left) with a plant wound-inoculated with spores of F. oxysporum (right) and grown in coco fiber substrate. Photo was taken 4 weeks after inoculation and shows stunting and yellowing of leaves. (I) Symptom of internal discoloration of the pith tissue in the upper 10 cm of the crown region of a plant grown indoors in coco fiber as a substrate and infected by F. oxysporum. Figures 2A, D, E, G reproduced from Can. J. Plant Pathol. 40(4) by permission.

Number of carbon atoms	Basic skeleton	Number of phenolic cycles	Class	Examples
6	C6	1	Simple phenols, Benzoquinones	Catechol, Hydroquinone, 2,6-Dimethoxybenzoquinone
7	C6-C1	1	Phenolic acids, Phenolic aldehydes	Gallic, salicylic acids
8	C6-C2	1	Acetophenones, Tyrosine derivatives, Phenylacetic acids	3-Acetyl-6-methoxybenzaldehyde, Tyrosol, p-Hydroxyphenylacetic acid, Homogentisic acid
9	C6-C3	1	Hydroxycinnamic acids, Allylbenzenes, Coumarins, Isocoumarins, Chromones	Caffeic, ferulic acids, Myristicin, Eugenol, Umbelliferone, aesculetin, Bergenon, Eugenin
10	C6-C4	1	Naphthoquinones	Juglone, Plumbagin
13	C6-C1-C6	2	Xanthonoids	Mangiferin
14	C6-C2-C6	2	Stilbenoids, Anthraquinones	Resveratrol, Emodin
15	C6-C3-C6	2	Chalconoids, Flavonoids, Isoflavonoids, Neoflavonoids	Quercetin, cyanidin, Genistein
16	C6-C4-C6	2	Halogenated algal phenolic compounds	Kaviol A, colpol
18	(C6-C3)2	2	Lignans, Neolignans	Pinoresinol, Eusiderin
30	(C6-C3-C6)2	4	Biflavonoids	Amentoflavone
many	(C6-C3)n, (C6)n, (C6-C3-C6)n	n > 12	Lignins, Catechol melanins, Flavolans (Condensed tannins), Polyphenolic proteins, Polyphenols	Raspberry ellagitannin, Tannic acid

Some phenols are germicidal and are used in formulating disinfectants.

Naturally occurring phenols - Wikipedia

en.wikipedia.org

ONCE THE PLANT PRODUCES ITS PHENOLS IN VITRO DIRT PLANT IT

 

[acespicoli]

Plant hormone​

Salicylic acid is a phenolic phytohormone, and is found in plants with roles in plant growth and development, photosynthesis, transpiration, and ion uptake and transport.[53] Salicylic acid is involved in endogenous signaling, mediating plant defense against pathogens.[54] It plays a role in the resistance to pathogens (i.e. systemic acquired resistance) by inducing the production of pathogenesis-related proteins and other defensive metabolites.[55] SA's defense signaling role is most clearly demonstrated by experiments which do away with it: Delaney et al. 1994, Gaffney et al. 1993, Lawton et al. 1995, and Vernooij et al. 1994 each use Nicotiana tabacum or Arabidopsis expressing nahG, for salicylate hydroxylase. Pathogen inoculation did not produce the customarily high SA levels, SAR was not produced, and no pathogenesis-related (PR) genes were expressed in systemic leaves. Indeed, the subjects were more susceptible to virulent – and even normally avirulent – pathogens.[53]

Exogenously, salicylic acid can aid plant development via enhanced seed germination, bud flowering, and fruit ripening, though too high of a concentration of salicylic acid can negatively regulate these developmental processes.[56]

The volatile methyl ester of salicylic acid, methyl salicylate, can also diffuse through the air, facilitating plant-plant communication.[57] Methyl salicylate is taken up by the stomata of the nearby plant, where it can induce an immune response after being converted back to salicylic acid.[58]

Signal transduction​

A number of proteins have been identified that interact with SA in plants, especially salicylic acid binding proteins (SABPs) and the NPR genes (nonexpressor of pathogenesis-related genes), which are putative receptors.[59

Salicylic acid - Wikipedia

en.wikipedia.org

 

[Mate Dave]

I had a little play around a while back making what I call 'fancy cuttings'.. I've mixed opinions..
I had some time I wanted to play with a 'bigger train-set'.

Cleaning probably has a few meanings to different people.

You 1st need to make some cleaning solution and a media that is sterile and will grow tiny plants. Start with some DD water and keep it in the dark but somewhere with a stable temperature.

Prepare a solution of DD water with a wetting agent, and 5% citric acid, froth it up to release the chemical foam. Use a pipette split the solution into 3 beakers discarding any foam from mixing. Heat this solution to 15c with the Hotplate stirrer machine.

This 'Bath' is a good way to cleanse, place plant material into solution and let it do it's thing for 15-20 mins.
Drain and use the other beakers in the same fashion as a rinse..

Or you could do multiple stages and have various washes using a variety of cleaning agents.

Diluteing DD to the ratio required add Sodium hyperchlorite 20% Ethanol 70% Erythromycin, 1 to 5 mg/mL. Hydrogen peroxide 3% and citric acid 5%.

Place cuttings into the 'Elution' and turn the Hotplate stirrer machine and wait 15 minutes.. Rinse repeat. Each time discard the bath water and add the cuttings to the second beaker and rinse them.

Each of the above elutants, have a critical contact time, even the citric acid will degrade plant materials after some time, exposure is therefore critical to success with these advanced steps..
Each process needs a rinse to remove any biofilm.

Cuttings can be treated to a UV B next and have dissections made from the very tips and placed into growing media, storage media, or Tempory emersion system (Rocker) or just propagated as normally and see what has happened in that stage.

I'd suggest to try do the basic wash, then go for a second round of the other cleaning agents getting used to tolerances. Keeping plant material as fresh as can be for storage.

Unless you can freeze the material you'll have a easier time doing what I have suggested and just cultivate cuttings normally as it's much easier than having to change storage media/vessels.

Practice making smaller and smaller dissections. Get used to filling up the "rocker" playing with various timings and liquid mediums.

I'm not sure that having to regrow plant parts from undifferentiated cells sat in agar is needed or time/cost effective for the home grower but certainly is the way of keeping viroid free parental stock for security.

 

[acespicoli]

I had a little play around a while back making what I call 'fancy cuttings'.. I've mixed opinions..
I had some time I wanted to play with a 'bigger train-set'.

Cleaning probably has a few meanings to different people.

You 1st need to make some cleaning solution and a media that is sterile and will grow tiny plants. Start with some DD water and keep it in the dark but somewhere with a stable temperature.

Prepare a solution of DD water with a wetting agent, and 5% citric acid froth it up to release the chemicals foam. Use a pipette split the solution into 3 beakers discarding any foam from mixing. Heat this to 15C.

This 'Bath' is a good way to cleanse place plant material into it and let it do it's thing for 15-20 mins.
drain and use the other beakers in the same fashion as a rinse.. or you could do multiple stages and have various washes using a variety of cleaning agents.

Diluteing DD to the ratio required add Sodium hyperchlorite 20% Ethanol 70% Erythromycin, 1 to 5 mg/mL. Hydrogen peroxide 3% and citric acid 5%.

Place cuttings into the solution and turn the machine on and wait 15 minutes.. Rinse repeat. Each time discard the bath water and add the cuttings to the second beaker and rinse them.

Each of the above elutants, have a critical contact time, even the citric acid will degrade plant materials after some time, exposure is therefore critical to success with these advanced steps..
Each process needs a rinse to remove any biofilm.

Cuttings can be treated to a UV B next and have dissections made from the very tips and placed into growing, media storage, or Tempory emersion system (Rocker) or just propagated as normally and see what has happened in that stage.

Id suggest to try do the basic wash and then go for a second round of
the other cleaning agents getting used to tolerances. Keeping plant material as fresh as can be for storage.

Unless you can freeze the material you'll have a easier time doing what I have suggested and just cultivate cuttings normally as it's much easier than having to change storage media..

Practice making smaller and smaller dissections. Get used to filling up the "rocker" playing with various timings and liquid mediums.

I'm not sure that having to regrow plant parts from undifferentiated cells sat in agar is needed or time/cost effective for the home grower but certainly is the way of keeping viroid free parental stock for security.

You raise some good points always good to look at, what are you trying to achieve?

That first cut you make takes all stem contaminants and introduces them the the interior of the plant.
Cut and recut after sterilizing less than the whole plant
Multi washes are a great idea or stepping thru each wash ingredient individually
We deal with bacteria fungus and virus so nice recipe you shared, thank you

 

[acespicoli]

COTTON'S TOTEby /u/cottonparachute​

This is my LED tote build! I'm using 100w LED chips powered by a 24v 15A power supply that I cranked all the way up to 30.5v. I had 2 boost converters at first, but one had heat issues boosting 24v to 30v and the other one went 2 weeks after. That was 2 months ago. Seems to work fine just having them connected in parallel on the power supply and I check on voltage, draw and temperature about once a week; so far so good. The LEDs are just generic 100w cool white (6500k). I also have warm white (3700k) COBs that I want to use 2 weeks into flower. I use 2 passive intakes on the bottom tote that are covered by mesh fan covers and a 90mm, 78cfm B-Blaster case fan in the back of the top tote.

Sterilite 16877404 25 Gallon Ultra Tote, True Blue lid & base with Titanium latches, 4-Pack
Sterilite

LOHAS® 100W LED Chip Cool White Bulb High Power Energy Saving Lamp Chip
$10.49
L LOHAS LED

AVAWO® DC 24V15A 360W Switching Power Supply Transformer Regulated for LED Strip Light, CCTV, Radio, Computer Project etc.
$20.98
AVAWO

LEDMO 5630 LED Strip Light, Warm White 16.4Ft 300LEDs Non-waterproof IP20 DC12V 60LEDs/m 25LM/LED, 2 times brightness than 5050 LED, LED Tape Light, LED Light Strip
LEDMO


Anyway as long as were dreaming, why not make it feasible for everyone ?
Disclaimer: Highly recommend not taking on electrical hacks, leave it to those qualified todo so
These can pose a fire hazard in non-competent hands and unconventional methods
Very neat looking, those COB led generate a great deal of heat... this is a DIY from the Space Bucket com
Use at your own risk would be much better with cool tubes or tape

 

[Mate Dave]

I wasn't trying to achieve anything but making life hard for myself. The wash just seems sensible. Was part of what I was learning at the time and I learned from it.

The 'Rocker' looks cool. It's a PITA to set and maintain liquid levels and humidity. I have rooted clones in plastic bags with no media.. Seems a bit superfluous ..

I was thinking it might have been a way to send genetics around at one time and leave plants whilst on holiday etc.

Thanks for the kind words.

I personally would sooner these processes done by laboratory technicians as it's hella boring mixing agar, sterilising and just not being in the grow room basically.

I got the stirrer to decarboxylate oli to test cannabinoids in TLC. Then I borrowed a 'Rocker' from University.

Toys like I said.

 

[acespicoli]

Tnelz​

Five days. Cheese clone given nothing but
coconut water @ 15 Mls and
aloe @ 5.
Also the root dip which is just beneficial bacteria and fungi from microbe life!

Rhodopseudomonas palustris
can use natural conductivity to pull electrons from minerals located remotely in soil and sediment while remaining at the surface, where they absorb the sunlight needed to produce energy.

 

[acespicoli]

Organisation of an apical meristem (growing tip)

1. Central zone
2. Peripheral zone
3. Medullary (i.e. central) meristem
4. Medullary tissue

There are two types of apical meristem tissue:
shoot apical meristem (SAM), which gives rise to organs like the leaves and flowers, and r
oot apical meristem (RAM), which provides the meristematic cells for future root growth.
SAM and RAM cells divide rapidly and are considered indeterminate,

in that they do not possess any defined end status.

• Shoot Apical Meristems
• Root apical meristem

In that sense, the meristematic cells are frequently compared to the stem cells in animals, which have an analogous behavior and function.

• Primary meristems
• Secondary meristems
• Apical meristems

The shoot apical meristem consists of four distinct cell groups:

• Stem cells
• The immediate daughter cells of the stem cells
• A subjacent organizing center
• Founder cells for organ initiation in surrounding regions

In multicellular organisms, stem cells are undifferentiated or partially differentiated cells that can change into various types of cells and proliferate indefinitely to produce more of the same stem cell. They are the earliest type of cell in a cell lineage.[1] They are found in both embryonic and adult organisms,

Cellular differentiation - Wikipedia

en.wikipedia.org

Power of one.
Orange tree embryos can sprout from a single somatic cell.
CREDITS: USHRL/ARS/USDA

In conclusion: Not all plant cells are totipotent, but under appropriate conditions certain cells may become totipotent. A cell (and only a single cell) can be considered as totipotent if it is able to autonomously develop into a whole plant via embryogenesis.

Pluripotency (biological compounds) - Wikipedia

en.wikipedia.org

Pluripotent is also described as something that has no fixed developmental potential, as in being able to differentiate into different cell types in the case of pluripotent stem cells.[1]

 

[Mate Dave]

These 'Purple Kush' root straight from the 'Bath water' if the plugs are new and the humidity is ok.
Don't even need any aloe or coconut water but I like it I don't use hormones or anything for cuttings. Only water. I don't really count days on the first to show feet either more looking as whole batches and strike rates. Changing pH adding gas recording the observations like a science experiment. "Disco"