Chap 2: Propagation of Cannabis, Part 2/2

Chapter 2: the Propagation of Cannabis.
Part 2/2----due to limit on post length

Making Cuttings

Cuttings of relatively young vegetative limbs 10 to 45 centimeters (4 to 18 inches) are made with a sharp knife or razor blade and immediately placed in a container of clean, pure water so the cut ends are well covered. It is essential that the cuttings be placed in water as soon as they are removed or a bubble of air (embolism) may enter the cut end and block the transpiration stream in the cutting, causing it to wilt. Cuttings made under water avoid the possibility of an embolism. If cuttings are exposed to the air they are cut again before being inserted into the rooting medium.

The medium should be warm and moist before cut tings are removed from the parental plant. Rows of holes are made in the rooting medium with a tapered stick, slightly larger in diameter than the cutting, leaving at least 10 centimeters (4 inches) between each hole. The cuttings are removed from the water, the end to be rooted treated with growth regulators and fungicides (such as Rootone F or Hormex), and each cutting placed in its hole. The cut end of the shoot is kept at least 10 centimeters (4 inches) from the bottom of the medium. The rooting medium is lightly tamped around the cutting, taking care not to scrape off the growth regulators. During the first few days the cuttings are checked frequently to make sure every thing is working properly. The cuttings are then watered with a mild nutrient solution once a day.

Hardening-off

The cuttings usually develop a good root system and will be ready to transplant in three to six weeks. At this time the hardening-off process begins, preparing the delicate cuttings for a life in bright sunshine. The cuttings are removed and transplanted to a sheltered spot such as a greenhouse until they begin to grow on their own. It is necessary to water them with a dilute nutrient solution or feed with finished compost as soon as the hardening-off process begins. Young roots are very tender and great care is necessary to avoid damage. When vegetative cuttings are placed outside under the prevailing photoperiod they will react accordingly. If it is not the proper time of the year for the cuttings to grow and mature properly (near harvest time, for example) or if it is too cold for them to be put out, then they may be kept in a vegetative condition by supplementing their light to increase daylength. Alternatively they may be induced to flower indoors under artificial conditions.

After shoots are selected and prepared for cloning, they are treated and placed in the rooting medium. Since the discovery in 1984 that auxins such as IAA stimulate the production of adventitious roots, and the subsequent discovery that the application of synthetic auxins such as NAA increase the rate of root production, many new techniques of treatment have appeared. It has been found that mixtures of growth regulators are often more effective than one alone. IAA and NAA aâ??e often combined with a small percentage of certain phenoxy compounds and fungicides in commercial preparations. Many growth regulators deteriorate rapidly, and fresh solutions are made up as needed. Treatments with vitamin B1 (thiamine) seem to help roots grow, but no inductive effect has been noticed. As soon as roots emerge, nutrients are necessary; the shoot cannot maintain growth for long on its own reserves. A complete complement of nutrients in the rooting medium certainly helps root growth; nitrogen is especially beneficial. Cuttings are extremely susceptible to fungus attack, and conditions conducive to rooting are also favorable to the growth of fungus. "Cap tan " is a long-lasting fungicide that is sometimes applied in powdered form along with growth regulators. This is done by rolling the basal end of the cutting in the powder before placing it in the rooting medium.

Oxygen and Rooting

The initiation and growth of roots depends upon atmospheric oxygen. If oxygen levels are low, shoots may fail to produce roots and rooting will certainly be inhibited. It is very important to select a light, well-aerated rooting medium. In addition to natural aeration from the atmosphere, rooting media may be enriched with oxygen (02) gas; enriched rooting solutions have been shown to increase rooting in many plant species. No threshold for damage by excess oxygenation has been determined, although excessive oxygenation could displace carbon dioxide which is also vital for proper root initiation and growth. If oxygen levels are low, roots will form only near the surface of the medium, whereas with adequate oxygen levels, roots will tend to form along the entire length of the implanted shoot, especially at the cut end.

Oxygen enrichment of rooting media is fairly simple. Since shoot cuttings must be constantly wetted to ensure proper rooting, aeration of the rooting media may be facilitated by aerating the water used in irrigation. Mist systems achieve this automatically because they deliver a fine mist (high in dissolved oxygen) to the leaves, from where much of it runs off into the soil, aiding rooting. Oxygen enrichment of irrigation water is accomplished by installing an aerator in the main water line so that atmospheric oxygen can be absorbed by the water. An increase in dissolved oxygen of only 20 parts per million may have a great influence on rooting. Aeration is a convenient way to add oxygen to water as it also adds carbon dioxide from the atmosphere. Air from a small pump or bottled oxygen may also be supplied directly to the rooting media through tiny tubes with pin holes, or through a porous stone such as those used to aerate aquariums.

Rooting Media

Water is a common medium for rooting. It is inexpensive, disperses nutrients evenly, and allows direct observation of root development. However, several problems arise. A water medium allows light to reach the submerged stem, delaying etiolation and slowing root growth. Water also promotes the growth of water molds and other fungi, sup ports the cutting poorly, and restricts air circulation to the young roots. In a well aerated solution, roots will appear in great profusion at the base of the stem, while in a poorly aerated or stagnant solution only a few roots will form at the surface, where direct oxygen exchange occurs. If rootings are made in pure water, the solution might be replaced regularly with tap water, which should contain sufficient oxygen for a short period. If nutrient solutions are used, a system is needed to oxygenate the solution. The nutrient solution does become concentrated by evaporation, and this is watched. Pure water is used to dilute rooting solutions and refill rooting containers.

Soil Treatment (read up to here)

Solid media provide anchors for cuttings, plenty of darkness to promote etiolation and root growth, and sufficient air circulation to the young roots. A high-quality soil with good drainage such as that used for seed germination is often used but the soil must be carefully sterilized to prevent the growth of harmful bacteria and fungus. A small amount of soil can easily be sterilized by spreading it out on a cookie sheet and heating it in an oven set at "low," approximately 820 C (180~ F), for thirty minutes. This kills most harmful bacteria and fungus as well as nematodes, in sects and most weed seeds. Overheating the soil will cause the breakdown of nutrients and organic complexes and the formation of toxic compounds. Large amounts of soil may be treated by chemical fumigants. Chemical fumigation avoids the breakdown of organic material by heat and may result in a better rooting mix. Formaldehyde is an excellent fungicide and kills some weed seeds, nematodes, and in sects. One gallon of commercial formalin (40% strength) is mixed with 50 gallons of water and slowly applied until each cubic foot of soil absorbs 2-4 quarts of solution. Small containers are sealed with plastic bags; large flats and plots are covered with polyethylene sheets. After 24 hours the seal is removed and the soil is allowed to dry for two weeks or until the odor of formaldehyde is no longer present. The treated soil is drenched with water prior to use. Fumigants such as formaldehyde, methyl bromide or other lethal gases are very dangerous and cultivators use them only outside with appropriate protection for themselves.

It is usually much simpler and safer to use an artificial sterile medium for rooting. Vermiculite and perlite are often used in propagation because of their excellent drain age and neutral pH (a balance between acidity and alkalinity). No sterilization is needed because both products are manufactured at high heat and contain no organic material. It has been found that a mixture of equal portions of medium and large grade vermiculite or perlite promotes the greatest root growth. This results from increased air circulation around the larger pieces. A weak nutrient solution, including micro-nutrients, is needed to wet the medium, because little or no nutrient material is supplied by these artificial media. Solutions are checked for pH and corrected to neutral with agricultural lime, dolomite lime, or oyster shell lime.

Layering

Layering is a process in which roots develop on a stem while it remains attached to, and nutritionally sup ported by the parent plant. The stem is then detached and the meristematic tip becomes a new individual, growing on its own roots, termed a layer. Layering differs from cutting because rooting occurs while the shoot is still attached to the parent. Rooting is initiated in layering by various stem treatments which interrupt the downward flow of photosynthates (products of photosynthesis) from the shoot tip. This causes the accumulation of auxins, carbohydrates and other growth factors. Rooting occurs in this treated area even though the layer remains attached to the parent. Water and mineral nutrients are supplied by the parent plant because only the phloem has been interrupted; the xylem tissues connecting the shoot to the parental roots remain intact (see illus. 1, page 29). In this manner, the propagator can overcome the problem of keeping a severed cutting alive while it roots, thus greatly in creasing the chances of success. Old woody reproductive stems that, as cuttings, would dry up and die, may be rooted by layering. Layering can be very time-consuming and is less practical for mass cloning of parental stock than removing and rooting dozens of cuttings. Layering, however, does give the small-scale propagator a high-success alternative which also requires less equipment than cuttings.

Techniques of Layering

Almost all layering techniques rely on the principle of etiolation. Both soil layering and air layering involve depriving the rooting portion of the stem of light, promoting rooting. Root-promoting substances and fungicides prove beneficial, and they are usually applied as a spray or powder. Root formation on layers depends on constant moisture, good air circulation and moderate temperatures at the site of rooting.

Soil Layering

Soil layering may be performed in several ways. The most common is known as tip layering. A long, supple vegetative lower limb is selected for layering, carefully bent so it touches the ground, and stripped of leaves and small shoots where the rooting is to take place. A narrow trench, 6 inches to a foot long and 2 to 4 inches deep, is dug parallel to the limb, which is placed along the bottom of the trench, secured with wire or wooden stakes, and buried with a small mound of soil. The buried section of stem may be girdled by cutting, crushed with a loop of wire, or twisted to disrupt the phloem tissue and cause the accumulation of substances which promote rooting. It may also be treated with growth regulators at this time.

Serpentine layering may be used to create multiple layers along one long limb. Several stripped sections of the limb are buried in separate trenches, making sure that at least one node remains above ground between each set of roots to allow shoots to develop. The soil surrounding the stem is kept moist at all times and may require wetting several times a day. A small stone or stick is inserted under each exposed section of stem to prevent the lateral shoot buds rotting from constant contact with the moist soil surface. Tip layers and serpentine layers may be started in small containers placed near the parental plant. Rooting usually begins within two weeks, and layers may be re moved with a sharp razor or clippers after four to six weeks. If the roots have become well established, transplanting may be difficult without damaging the tender root system. Shoots on layers continue to grow under the same conditions as the parent, and less time is needed for the clone to acclimatize or harden-off and begin to grow on its own than with cuttings.

In air layering, roots form on the aerial portions of stems that have been girdled, treated with growth regulators, and wrapped with moist rooting media. Air layering is an ancient form of propagation, possibly invented by the Chinese. The ancient technique of goo tee uses a ball of clay or soil plastered around a girdled stem and held with a wrap of fibers. Above this is suspended a small container of water (such as a bamboo section) with a wick to the wrapped gootee; this way the gootee remains moist.

The single most difficult problem with air layers is the tendency for them to dry out quickly. Relatively small amounts of rooting media are used, and the position on aerial parts of the plant exposes them to drying winds and sun. Many wraps have been tried, but the best seems to be clear polyethylene plastic sheeting which allows oxygen to enter and retains moisture well. Air layers are easiest to make in greenhouses where humidity is high, but they may also be used outside as long as they are kept moist and donâ??t freeze. Air layers are most useful to the amateur propagator and breeder because they take up little space and allow the efficient cloning of many individuals.

Making an Air Layer

A recently sexed young limb 3-10 mm (1/8 to 3/8 inch) in diameter is selected. The site of the layer is usually a spot 30 centimeters (12 inches) or more from the limb tip. Unless the stem is particularly strong and woody, it is splinted by positioning a 30 centimeter (12 inch) stick of approximately the same diameter as the stem to be layered along the bottom edge of the stem. This splint is tied in place at both ends with a piece of elastic plant-tie tape. This enables the propagator to handle the stem more confidently. An old, dry Cannabis stem works well as a splint. Next, the stem is girdled between the two ties with a twist of wire or a diagonal cut. After girdling, the stem is sprayed or dusted with a fungicide and growth regulator, surrounded with one or two handfuls of unmilled sphagnum moss, and wrapped tightly with a small sheet of clear polyethylene film (4-6 mil). The film is tied securely at each end, tightly enough to make a waterproof seal but not so tight that the phloem tissues are crushed. If the phloem is crushed, compounds necessary for rooting will accumulate outside of the medium and rooting will be slowed. Plastic floristâ??s tape or electricianâ??s tape works well for sealing air layers. Although polyethylene film retains moisture well, the moss will dry out eventually and must be remoistened periodically. Unwrapping each layer is impractical and would disturb the roots, so a hypodermic syringe is used to inject water, nutrients, fungicides, and growth regulators. If the layers become too wet the limb rots. Layers are checked regularly by injecting water until it squirts out and then very lightly squeezing the medium to remove any extra water. Heavy layers on thin limbs are supported by tying them to a large adjacent limb or a small stick anchored in the ground. Rooting begins within two weeks and roots will be visible through the clear plastic within four weeks. When the roots appear adequately developed, the layer is removed, carefully unwrapped, and transplanted with the moss and the splint intact. The layer is watered well and placed in a shady spot for a few days to allow the plant to harden-off and adjust to living on its own root system. It is then placed in the open. In hot weather, large leaves are removed from the shoot before removing the layer to prevent excessive transpiration and wilting.

Layers develop fastest just after sexual differentiation. Many layers may be made of staminate plants in order to save small samples of them for pollen collection and to conserve space. By the time the pollen parents begin to flower profusely, the layers will be rooted and may be cut and removed to an isolated area. Layers taken from pistil late plants are used for breeding, or saved and cloned for the following season.

Layers often seem rejuvenated when they are re moved from the parent plant and begin to be supported by their own root systems. This could mean that a clone will continue to grow longer and mature later than its parent under the same conditions. Layers removed from old or seeded parents will continue to produce new calyxes and pistils instead of completing the life cycle along with the parents. Rejuvenated layers are useful for off-season seed production.

Grafting

Intergeneric grafts between Cannabis and Humulus (hops) have fascinated researchers and cultivators for decades. Warmke and Davidson (1943) claimed that Humbles tops grafted upon Cannabis roots produced ". . . as much drug as leaves from intact hemp plants, even though leaves from intact hop plants are completely nontoxic." According to this research, the active ingredient of Cannabis was being produced in the roots and transported across the graft to the Humulus tops. Later research by Crombie and Crombie (1975) entirely disproves this theory. Grafts were made between high and low THC strains of Cannabis as well as intergeneric grafts between Cannabis and Humulus, Detailed chromatographic analysis was performed on both donors for each graft and their control populations. The results showed ". . . no evidence of transport of inter mediates or factors critical to cannabinoid formation across the grafts."

Grafting of Cannabis is very simple. Several seedlings can be grafted together into one to produce very interesting specimen plants. One procedure starts by planting one seed ling each of several separate strains close together in the same container, placing the stock (root plant) for the cross in the center of the rest. When the seedlings are four weeks old they are ready to be grafted. A diagonal cut is made approximately half-way through the stock stem and one of the scion (shoot) seedlings at the same level. The cut portions are slipped together such that the inner cut surfaces are touching. The joints are held with a fold of cellophane tape. A second scion from an adjacent seedling may be grafted to the stock higher up the stem. After two weeks, the unwanted portions of the grafts are cut away. Eight to twelve weeks are needed to complete the graft, and the plants are maintained in a mild environment at all times. As the graft takes, and the plant begins to grow, the tape falls off.

Pruning

Pruning techniques are commonly used by Cannabis cultivators to limit the size of their plants and promote branching. Several techniques are available, and each has its advantages and drawbacks. The most common method is meristem pruning or stem tip removal. In this case the growing tip of the main stalk or a limb is removed at approximately the final length desired for the stalk or limb. Below the point of removal, the next pair of axial growing tips begins to elongate and form two new limbs. The growth energy of one stem is now divided into two, and the diffusion of growth energy results in a shorter plant which spreads horizontally.

Auxin produced in the tip meristem travels down the stem and inhibits branching. When the meristem is re moved, the auxin is no longer produced and branching may proceed uninhibited. Plants that are normally very tall and stringy can be kept short and bushy by meristem pruning. Removing meristems also removes the newly formed tissues near the meristem that react to changing environmental stimuli and induce flowering. Pruning during the early part of the growth cycle will have little effect on flowering, but plants that are pruned late in life, supposedly to promote branching and floral growth, will often flower late or fail to flower at all. This happens because the meristemic tissue responsible for sensing change has been removed and the plant does not measure that it is the time of the year to flower. Plants will usually mature fastest if they are allowed to grow and develop without interference from pruning. If late maturation of Cannabis is desired, then extensive pruning may work to delay flowering. This is particularly applicable if a staminate plant from an early maturing strain is needed to pollinate a late-maturing pistil late plant. The staminate plant is kept immature until the pistillate plant is mature and ready to be pollinated. When the pistillate plant is receptive, the staminate plant is allowed to develop flowers and release pollen.

Other techniques are available for limiting the size and shape of a developing Cannabis plant without removing meristematic tissues. Trellising is a common form of modification and is achieved in several ways. In many cases space is available only along a fence or garden row. Posts 1 to 2 meters (3 to 6 feet) long may be driven into the ground 1 to 3 meters (3 to 10 feet) apart and wires stretched between them at 30 to 45 centimeters (12 to 18 inches) intervals, much like a wire fence or grape trellis. Trellises are ideally oriented on an east-west axis for maxi mum sun exposure. Seedlings or pistillate clones are placed between the posts, and as they grow they are gradually bent and attached to the wire. The plant continues to grow upward at the stem tips, but the limbs are trained to grow horizontally. They are spaced evenly along the wires by hooking the upturned tips under the wire when they are 15 to 30 centimeters (6 to 12 inches) long. The plant grows and spreads for some distance, but it is never allowed to grow higher than the top row of wire. When the plant be gins to flower, the floral clusters are allowed to grow up ward in a row from the wire where they receive maximum sun exposure. The floral clusters are supported by the wire above them, and they are resistant to weather damage. Many cultivators feel that trellised plants, with increased sun exposure and meristems intact, produce a higher yield than freestanding unpruned or pruned plants. Other growers feel that any interference with natural growth patterns limits the ultimate size and yield of the plant.

Another method of trellising is used when light exposure is especially crucial, as with artificial lighting systems. Plants are placed under a horizontal or slightly slanted flat sheet of 2 to 5 centimeters (1 to 2 inches) poultry netting which is suspended on a frame 30 to 60 centimeters (12 to 24 inches) from the soil surface perpendicular to the direction of incoming light or to the lowest path of the sun. The seedlings or clones begin to grow through the netting al-â?? most immediately, and the meristems are pushed back down under the netting, forcing them to grow horizon tally outward. Limbs are trained so that the mature plant will cover the entire frame evenly. Once again, when the plant begins to flower, the floral clusters are allowed to grow upward through the wire as they reach for the light. This might prove to be a feasible commercial cultivation technique, since the flat beds of floral clusters could be mechanically harvested. Since no meristem tissues are re moved, growth and maturation should proceed on schedule. This system also provides maximum light exposure for all the floral clusters, since they are growing from a plane perpendicular to the direction of light.

Sometimes limbs are also tied down, or crimped and bent to limit height and promote axial growth without meristem removal. This is a particularly useful technique for greenhouse cultivation, where plants often reach the roof or walls and burn or rot from the intense heat and condensation of water on the inside of the greenhouse. To prevent rotting and burning while leaving enough room for floral clusters to form, the limbs are bent at least 60 centimeters (24 inches) beneath the roof of the green house. Tying plants over allows more light to strike the plant, promoting axial growth. Crimping stems and bending them over results in more light exposure as well as inhibiting the flow of auxin down the stem from the tip. Once again, as with meristem removal, this promotes axial growth.

Limbing is another common method of pruning Cannabis plants. Many small limbs will usually grow from the bottom portions of the plant, and due to shading they re main small and fail to develop large floral clusters. If these atrophied lower limbs are removed, the plant can devote more of its floral energies to the top parts of the plant with the most sun exposure and the greatest chance of pollination. The question arises of whether removing entire limbs constitutes a shock to the growing plant, possibly limiting its ultimate size. It seems in this case that shock is minimized by removing entire limbs, including proportional amounts of stems, leaves, meristems, and flowers; this probably results in less metabolic imbalance than if only flowers, leaves, or meristems were removed. Also, the lower limbs are usually very small and seem of little significance in the metabolism of the total plant. In large plants, many limbs near the central stalk also become shaded and atrophied and these are also sometimes removed in an effort to increase the yield of large floral clusters on the sunny exterior margins.

Leafing is one of the most misunderstood techniques of drug Cannabis cultivation. In the mind of the cultivator, several reasons exist for removing leaves. Many feel that large shade leaves draw energy from the flowering plant, and therefore the flowering clusters will be smaller. It is felt that by removing the leaves, surplus energy will be available, and large floral clusters will be formed. Also, some feel that inhibitors of flowering, synthesized in the leaves during the long noninductive days of summer, may be stored in the older leaves that were formed during the noninductive photoperiod. Possibly, if these inhibitor-laden leaves are removed, the plant will proceed to flower, and maturation will be accelerated. Large leaves shade the inner portions of the plant, and small atrophied floral clusters may begin to develop if they receive more light.

In actuality, few if any of the theories behind leafing give any indication of validity. Indeed, leafing possibly serves to defeat its original purpose. Large leaves have a definite function in the growth and development of Cannabis. Large leaves serve as photosynthetic factories for the production of sugars and other necessary growth sub stances. They also create shade, but at the same time they are collecting valuable solar energy and producing foods that will be used during the floral development of the plant. Premature removal of leaves may cause stunting, because the potential for photosynthesis is reduced. As these leaves age and lose their ability to carry on photo synthesis they turn chlorotie (yellow) and fall to the ground. In humid areas care is taken to remove the yellow or brown leaves, because they might invite attack by fungus. During chlorosis the plant breaks down substances, such as chlorophylls, and translocates the molecular components to a new growing part of the plant, such as the flowers. Most Cannabis plants begin to lose their larger leaves when they enter the flowering stage, and this trend continues until senescence. It is more efficient for the plant to reuse the energy and various molecular components of existing chlorophyll than to synthesize new chlorophyll at the time of flowering. During flowering this energy is needed to form floral clusters and ripen seeds.

Removing large amounts of leaves may interfere with the metabolic balance of the plant. If this metabolic change occurs too late in the season it could interfere with floral development and delay maturation. If any floral inhibitors are removed, the intended effect of accelerating flowering will probably be counteracted by metabolic upset in the plant. Removal of shade leaves does facilitate more light reaching the center of the plant, but if there is not enough food energy produced in the leaves, the small internal floral clusters will probably not grow any larger. Leaf removal may also cause sex reversal resulting from a metabolic change.

If leaves must be removed, the petiole is cut so that at least an inch remains attached to the stalk. Weaknesses in the limb axis at the node result if the leaves are pulled off at the abscission layer while they are still green. Care is taken to see that the shriveling petiole does not invite fungus attack.

It should be remembered that, regardless of strain or environmental conditions, the plant strives to reproduce, and reproduction is favored by early maturation. This produces a situation where plants are trying to mature and reproduce as fast as possible. Although the purpose of leafing is to speed maturation, disturbing the natural progressive growth of a plant probably interferes with its rapid development.

Cannabis grows largest when provided with plentiful nutrients, sunlight, and water and left alone to grow and mature naturally. It must be remembered that any alteration of the natural life cycle of Cannabis will affect productivity. Imaginative combinations and adaptations of propagation techniques exist, based on specific situations of cultivation. Logical choices are made to direct the natural growth cycle of Cannabis to favor the timely maturation of those products sought by the cultivator, without sacrificing seed or clone production.