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Marijuana Botany
An Advanced Study: The Propagation and Breeding of Distinctive Cannabis
by Robert Connell Clarke
Chapter 2 - Propagation of Cannabis
"Make the most of the Indian Hemp Seed and sow it every where."
- George Washington
Sexual versus Asexual Propagation
Cannabis can be propagated either sexually or asexually. Seeds are the
result of sexual propagation. Because sexual propagation involves the
recombination of genetic material from two parents we expect to observe
variation among seedlings and offspring with characteristics differing from
those of the parents. Vegetative methods of propagation (cloning) such as
cuttage, layerage, or division of roots are asexual and allow exact
replication of the parental plant without genetic variation. Asexual
propagation, in theory, allows strains to be preserved unchanged through
many seasons and hundreds of individuals.
When the difference between sexual and asexual propagation is well
understood then the proper method can be chosen for each situation. The
unique characteristics of a plant result from the combination of genes in
chromosomes present in each cell, collectively known as the genotype of that
individual. The expression of a genotype, as influenced by the environment,
creates a set of visible characteristics that we collectively term the
phenotype. The function of propagation is to preserve special genotypes by
choosing the proper technique to ensure replication of the desired
characteristics.
If two clones from a pistillate Cannabis plant are placed in differing
environments, shade and sun for in stance, their genotypes will remain
identical. However, the clone grown in the shade will grow tall and slender
and mature late, while the clone grown in full sun will remain short and
bushy and mature much earlier.
Sexual Propagation
Sexual propagation requires the union of staminate pollen and pistillate
ovule, the formation of viable seed, and the creation of individuals with
newly recombinant genotypes. Pollen and ovules are formed by reduction
divisions (meiosis) in which the 10 chromosome pairs fail to replicate, so
that each of the two daughter-cells contains one-half of the chromosomes
from the mother cell. This is known as the haploid (in) condition where in =
10 chromosomes. The diploid condition is restored upon fertilization
resulting in diploid (2n) individuals with a haploid set of chromosomes from
each parent. Offspring may resemble the staminate, pistillate, both, or
neither parent and considerable variation in offspring is to be expected.
Traits may be controlled by a single gene or a combination of genes,
resulting in further potential diversity.
The terms homozygous and heterozygous are useful in describing the
genotype of a particular plant. If the genes controlling a trait are the
same on one chromosome as those on the opposite member of the chromosome
pair (homologous chromosomes), the plant is homozygous and will "breed true"
for that trait if self-pollinated or crossed with an individual of identical
genotype for that trait. The traits possessed by the homozygous parent will
be transmitted to the offspring, which will resemble each other and the
parent. If the genes on one chromosome differ from the genes on its
homologous chromosome then the plant is termed heterozygous; the resultant
offspring may not possess the parental traits and will most probably differ
from each other. Imported Cannabis strains usually exhibit great seedling
diversity for most traits and many types will be discovered.
To minimize variation in seedlings and ensure preservation of desirable
parental traits in offspring, certain careful procedures are followed as
illustrated in Chapter III. The actual mechanisms of sexual propagation and
seed production will be thoroughly explained here.
The Life Cycle and Sinsemilla Cultivation
A wild Cannabis plant grows from seed to a seedling, to a prefloral
juvenile, to either pollen- or seed-bearing adult, following the usual
pattern of development and sexual reproduction. Fiber and drug production
both interfere with the natural cycle and block the pathways of inheritance.
Fiber crops are usually harvested in the juvenile or prefloral stage, before
viable seed is produced, while sinsemilla or seedless marijuana cultivation
eliminates pollination and subsequent seed production. In the case of
cultivated Cannabis crops, special techniques must be used to produce viable
seed for the following year without jeopardizing the quality of the final
product.
Modern fiber or hemp farmers use commercially produced high fiber content
strains of even maturation. Monoecious strains are often used because they
mature more evenly than dioecious strains. The hemp breeder sets up test
plots where phenotypes can be recorded and controlled crosses can be made. A
farmer may leave a portion of his crop to develop mature seeds which he
collects for the following year. If a hybrid variety is grown, the offspring
will not ail resemble the parent crop and desirable characteristics may be
lost.
Growers of seeded marijuana for smoking or hashish production collect
vast quantities of seeds that fall from the flowers during harvesting,
drying, and processing. A mature pistillate plant can produce tens of
thousands of seeds if freely pollinated. Sinsemilla marijuana is grown by
removing all the staminate plants from a patch, eliminating every pollen
source, and allowing the pistillate plants to produce massive clusters of
unfertilized flowers.
Various theories have arisen to explain the unusually potent psychoactive
properties of unfertilized Cannabis. In general these theories have as their
central theme the extraordinarily long, frustrated struggle of the
pistillate plant to reproduce, and many theories are both twisted and
romantic. What actually happens when a pistillate plant remains unfertilized
for its entire life and how this ultimately affects the cannabinoid (class
of molecules found only in Cannabis) and terpene (a class of aromatic
organic compounds) levels remains a mystery. It is assumed, how ever, that
seeding cuts the life of the plant short and THC (tetrahydrocannabinol the
major psychoactive compound in Cannabis) does not have enough time to
accumulate. Hormonal changes associated with seeding definitely affect all
metabolic processes within the plant including cannabinoid biosynthesis. The
exact nature of these changes is unknown but probably involves imbalance in
the enzymatic systems controlling cannabinoid production. Upon fertilization
the plant’s energies are channeled into seed production instead of increased
resin production. Sinsemilla plants continue to produce new floral clusters
until late fail, while seeded plants cease floral production. It is also
suspected that capitate-stalked trichome production might cease when the
calyx is fertilized. If this is the case, then sinsemilla may be higher in
THC because of uninterrupted floral growth, trichome formation and
cannabinoid production. What is important with respect to propagation is
that once again the farmer has interfered with the life cycle and no
naturally fertilized seeds have been produced.
The careful propagator, however, can produce as many seeds of pure types
as needed for future research without risk of pollinating the precious crop.
Staminate parents exhibiting favorable characteristics are reproductively
isolated while pollen is carefully collected and applied to only selected
flowers of the pistillate parents.
Many cultivators overlook the staminate plant, considering it useless if
not detrimental. But the staminate plant contributes half of the genotype
expressed in the offspring. Not only are staminate plants preserved for
breeding, but they must be allowed to mature, uninhibited, until their
phenotypes can be determined and the most favorable individuals selected.
Pollen may also be stored for short periods of time for later breeding.
Biology of Pollination
Pollination is the event of pollen landing on a stigmatic surface such as
the pistil, and fertilization is the union of the staminate chromosomes from
the pollen with the pistillate chromosomes from the ovule.
Pollination begins with dehiscence (release of pollen) from staminate
flowers. Millions of pollen grains float through the air on light breezes,
and many land on the stigmatic surfaces of nearby pistillate plants. If the
pistil is ripe, the pollen grain will germinate and send out a long pollen
tube much as a seed pushes out a root. The tube contains a haploid (in)
generative nucleus and grows downward toward the ovule at the base of the
pistils. When the pollen tube reaches the ovule, the staminate haploid
nucleus fuses with the pistillate haploid nucleus and the diploid condition
is restored. Germination of the pollen grain occurs 15 to 20 minutes after
contact with the stigmatic surface (pistil); fertilization may take up to
two days in cooler temperatures. Soon after fertilization, the pistils
wither away as the ovule and surrounding calyx begin to swell. If the plant
is properly watered, seed will form and sexual reproduction is complete. It
is crucial that no part of the cycle be interrupted or viable seed will not
form. If the pollen is subjected to extremes of temperature, humidity, or
moisture, it will fail to germinate, the pollen tube will die prior to
fertilization, or the embryo will be unable to develop into a mature seed.
Techniques for successful pollination have been designed with all these
criteria in mind.
Controlled versus Random Pollinations
The seeds with which most cultivators begin represent varied genotypes
even when they originate from the same floral cluster of marijuana, and not
all of these genotypes will prove favorable. Seeds collected from imported
shipments are the result of totally random pollinations among many
genotypes. If elimination of pollination was at tempted and only a few seeds
appear, the likelihood is very high that these pollinations were caused by a
late flowering staminate plant or a hermaphrodite, adversely affecting the
genotype of the offspring. Once the offspring of imported strains are in the
hands of a competent breeder, selection and replication of favorable
phenotypes by controlled breeding may begin. Only one or two individuals out
of many may prove acceptable as parents. If the cultivator allows random
pollination to occur again, the population not only fails to improve, it may
even degenerate through natural and accidental selection of unfavorable
traits. We must therefore turn to techniques of controlled pollination by
which the breeder attempts to take control and deter mine the genotype of
future offspring.
Data Collection
Keeping accurate notes and records is a key to successful plant-breeding.
Crosses among ten pure strains (ten staminate and ten pistillate parents)
result in ten pure and ninety hybrid crosses. It is an endless and
inefficient task to attempt to remember the significance of each little
number and colored tag associated with each cross. The well organized
breeder will free himself from this mental burden and possible confusion by
entering vital data about crosses, phenotypes, and growth conditions in a
system with one number corresponding to each member of the population.
The single most important task in the proper collection of data is to
establish undeniable credibility. Memory fails, and remembering the steps
that might possibly have led to the production of a favorable strain does
not constitute the data needed to reproduce that strain. Data is always
written down; memory is not a reliable record. A record book contains a
numbered page for each plant, and each separate cross is tagged on the
pistillate parent and recorded as follows: "seed of pistillate parent X
pollen or staminate parent." Also the date of pollination is included and
room is left for the date of seed harvest. Samples of the parental plants
are saved as voucher specimens for later characterization and analysis.
Pollination Techniques
Controlled hand pollination consists of two basic steps: collecting
pollen from the anthers of the staminate parent and applying pollen to the
receptive stigmatic surfaces of the pistillate parent. Both steps are
carefully con trolled so that no pollen escapes to cause random
pollinations. Since Cannabis is a wind-pollinated species, enclosures are
employed which isolate the ripe flowers from wind, eliminating pollination,
yet allowing enough light penetration and air circulation for the pollen and
seeds to develop without suffocating. Paper and very tightly woven cloth
seem to be the most suitable materials. Coarse cloth allows pollen to escape
and plastic materials tend to collect transpired water and rot the flowers.
Light-colored opaque or translucent reflective materials remain cooler in
the sun than dark or transparent materials, which either absorb solar heat
directly or create a greenhouse effect, heating the flowers inside and
killing the pollen. Pollination bags are easily constructed by gluing
together vegetable parchment (a strong breathable paper for steaming
vegetables) and clear nylon oven bags (for observation windows) with silicon
glue. Breathable synthetic fabrics such as Gore-Tex are used with great
success. Seed production requires both successful pollination and
fertilization, so the conditions inside the enclosures must remain suitable
for pollen-tube growth and fertilization. It is most convenient and
effective to use the same enclosure to collect pollen and apply it, reducing
contamination during pollen transfer. Controlled "free" pollinations may
also be made if only one pollen parent is allowed to remain in an isolated
area of the field and no pollinations are caused by hermaphrodites or
late-maturing staminate plants. If the selected staminate parent drops
pollen when there are only a few primordial flowers on the pistillate seed
parent, then only a few seeds will form in the basal flowers and the rest of
the flower cluster will be seedless. Early fertilization might also help fix
the sex of the pistillate plant, helping to prevent hermaphrodism. Later,
hand pollinations can be performed on the same pistillate parent by removing
the early seeds from each limb to be re-pollinated, so avoiding confusion.
Hermaphrodite or monoecious plants may be isolated from the remainder of the
population and allowed to freely self-pollinate if pure-breeding offspring
are desired to preserve a selected trait. Selfed hermaphrodites usually give
rise to hermaphrodite offspring.
Pollen may be collected in several ways. If the propagator has an
isolated area where staminate plants can grow separate from each other to
avoid mutual contamination and can be allowed to shed pollen without
endangering the remainder of the population, then direct collection may be
used. A small vial, glass plate, or mirror is held beneath a recently-opened
staminate flower which appears to be releasing pollen, and the pollen is
dislodged by tap ping the anthers. Pollen may also be collected by placing
whole limbs or clusters of staminate flowers on a piece of paper or glass
and allowing them to dry in a cool, still place. Pollen will drop from some
of the anthers as they dry, and this may be scraped up and stored for a
short time in a cool, dark, dry spot. A simple method is to place the open
pollen vial or folded paper in a larger sealable container with a dozen or
more fresh, dry soda crackers or a cup of dry white rice. The sealed
container is stored in the refrigerator and the dry crackers or rice act as
a desiccant, absorbing moisture from the pollen.
Any breeze may interfere with collection and cause contamination with
pollen from neighboring plants. Early morning is the best time to collect
pollen, as it has not been exposed to the heat of the day. All equipment
used for collection, including hands, must be cleaned before continuing to
the next pollen source. This ensures protection of each pollen sample from
contamination with pollen from different plants.
Staminate flowers will often open several hours before the onset of
pollen release. If flowers are collected at this time they can be placed in
a covered bottle where they will open and release pollen within two days. A
carefully sealed paper cover allows air circulation, facilitates the release
of pollen, and prevents mold.
Both of the previously described methods of pollen collection are
susceptible to gusts of wind, which may cause contamination problems if the
staminate pollen plants grow at all close to the remaining pistillate
plants. There fore, a method has been designed so that controlled pollen
collection and application can be performed in the same area without the
need to move staminate plants from their original location. Besides the
advantages of convenience, the pollen parents mature under the same
conditions as the seed parents, thus more accurately expressing their
phenotypes.
The first step in collecting pollen is, of course, the selection of a
staminate or pollen parent. Healthy individuals with well-developed clusters
of flowers are chosen. The appearance of the first staminate primordia or
male sex signs often brings a feeling of panic ("stamenoia") to the
cultivator of seedless Cannabis, and potential pollen parents are
prematurely removed. Staminate primordia need to develop from one to five
weeks before the flowers open and pollen is released. During this period the
selected pollen plants are carefully watched, daily or hourly if necessary,
for developmental rates vary greatly and pollen may be released quite early
in some strains. The remaining staminate plants that are unsuitable for
breeding are destroyed and the pollen plants specially labeled to avoid
confusion and extra work.
As the first flowers begin to swell, they are removed prior to pollen
release and destroyed. Tossing them on the ground is ineffective because
they may release pollen as they dry. When the staminate plant enters its
full floral condition and more ripe flowers appear than can be easily
controlled, limbs with the most ripe flowers are chosen. It is usually
safest to collect pollen from two limbs for each intended cross, in case one
fails to develop. If there are ten prospective seed parents, pollen from
twenty limbs on the pollen parent is collected. In this case, the twenty
most flowered limb tips are selected and all the remaining flowering
clusters on the plant are removed to prevent stray pollinations. Large
leaves are left on the remainder of the plant but are removed at the limb
tips to minimize condensation of water vapor released inside the enclosure.
The portions removed from the pollen parent are saved for later analysis and
phenotype characterization.
The pollination enclosures are secured and the plant is checked for any
shoots where flowers might develop outside the enclosure. The completely
open enclosure is slipped over the limb tip and secured with a tight but
stretchable seal such as a rubber band, elastic, or plastic plant tie-tape
to ensure a tight seal and prevent crushing of the vascular tissues of the
stem. String and wire are avoided. If enclosures are tied to weak limbs they
may be supported; the bags will also remain cooler if they are shaded. Hands
are always washed before and after handling each pollen sample to prevent
accidental pollen transfer and contamination.
Enclosures for collecting and applying pollen and preventing stray
pollination are simple in design and construction. Paper bags make
convenient enclosures. Long narrow bags such as light-gauge quart-bottle
bags, giant popcorn bags or bakery bags provide a convenient shape for
covering the limb tip. The thinner the paper used the more air circulation
is allowed, and the better the flowers will develop. Very thick paper or
plastic bags are never used. Most available bags are made with water soluble
glue and may come apart after rain or watering. All seams are sealed with
waterproof tape or silicon glue and the bags should not be handled when wet
since they tear easily. Bags of Gore-Tex cloth or vegetable parchment will
not tear when wet. Paper bags make labeling easy and each bag is marked in
waterproof ink with the number of the individual pollen parent, the date and
time the enclosure was secured, and any useful notes. Room is left to add
the date of pollen collection and necessary information about the future
seed parent it will pollinate.
Pollen release is fairly rapid inside the bags, and after two days to a
week the limbs may be removed and dried in a cool dark place, unless the
bags are placed too early or the pollen parent develops very slowly. To
inspect the progress of pollen release, a flashlight is held behind the bag
at night and the silhouettes of the opening flowers are easily seen. In some
cases, clear nylon windows are in stalled with silicon glue for greater
visibility. When flowering is at its peak and many flowers have just opened,
collection is completed, and the limb, with its bag attached, is cut. If the
limb is cut too early, the flowers will not have shed any pollen; if the bag
remains on the plant too long, most of the pollen will be dropped inside the
bag where heat and moisture will destroy it. When flowering is at its peak,
millions of pollen grains are released and many more flowers will open after
the limbs are collected. The bags are collected early in the morning before
the sun has time to heat them up. The bags and their contents are dried in a
cool dark place to avoid mold and pollen spoilage. If pollen becomes moist,
it will germinate and spoil, therefore dry storage is imperative.
After the staminate limbs have dried and pollen re lease has stopped, the
bags are shaken vigorously, allowed to settle, and carefully untied. The
limbs and loose flowers are removed, since they are a source of moisture
that could promote mold growth, and the pollen bags are re sealed. The bags
may be stored as they are until the seed parent is ready for pollination, or
the pollen may be re moved and stored in cool, dry, dark vials for later use
and hand application. Before storing pollen, any other plant parts present
are removed with a screen. A piece of fuel filter screening placed across
the top of a mason jar works well, as does a fine-mesh tea strainer.
Now a pistillate plant is chosen as the seed parent. A pistillate flower
cluster is ripe for fertilization so long as pale, slender pistils emerge
from the calyxes. Withered, dark pistils protruding from swollen, resin
encrusted calyxes are a sign that the reproductive peak has long passed.
Cannabis plants can be successfully pollinated as soon as the first
primordia show pistils and until just before harvest, but the largest yield
of uniform, healthy seeds is achieved by pollinating in the peak floral
stage. At this time, the seed plant is covered with thick clusters of white
pistils. Few pistils are brown and withered, and resin production has just
begun. This is the most receptive time for fertilization, still early in the
seed plant’s life, with plenty of time remaining for the seeds to mature.
Healthy, well flowered lower limbs on the shaded side of the plant are
selected. Shaded buds will not heat up in the bags as much as buds in the
hot sun, and this will help protect the sensitive pistils. When possible,
two terminal clusters of pistillate flowers are chosen for each pollen bag.
In this way, with two pollen bags for each seed parent and two clusters of
pistillate flowers for each bag, there are four opportunities to perform the
cross successfully. Remember that production of viable seed requires
successful pollination, fertilization and embryo development. Since
interfering with any part of this cycle precludes seed development,
fertilization failure is guarded against by duplicating all steps.
Before the pollen bags are used, the seed parent information is added to
the pollen parent data. Included is the number of the seed parent, the date
of pollination, and any comments about the phenotypes of both parents. Also,
for each of the selected pistillate clusters, a tag containing the same
information is made and secured to the limb below the closure of the bag. A
warm, windless evening is chosen for pollination so the pollen tube has time
to grow before sunrise. After removing most of the shade leaves from the
tips of the limbs to be pollinated, the pollen is tapped away from the mouth
of the bag. The bag is then carefully opened and slipped over two inverted
limb tips, taking care not to release any pollen, and tied securely with an
expandable band. The bag is shaken vigorously, so the pollen will be evenly
dispersed throughout the bag, facilitating complete pollination. Fresh bags
are sometimes used, either charged with pollen prior to being placed over
the limb tip, or injected with pollen, using a large syringe or atomizer,
after the bag is placed. However, the risk of accidental pollination with
injection is higher.
If only a small quantity of pollen is available it may be used more
sparingly by diluting with a neutral powder such as flour before it is used.
When pure pollen is used, many pollen grains may land on each pistil when
only one is needed for fertilization. Diluted pollen will go further and
still produce high fertilization rates. Diluting 1 part pollen with 10 to
100 parts flour is common. Powdered fungicides can also be used since this
helps retard the growth of molds in the maturing, seeded, floral clusters.
The bags may remain on the seed parent for sometime; seeds usually begin
to develop within a few days, buttheir development will be retarded by the
bags. The propagator waits three full sunny days, then carefully removes and
sterilizes or destroys the bags. This way there is little chance of stray
pollination. Any viable pollen that failed to pollinate the seed parent will
germinate in the warm moist bag and die within three days, along with many
of the unpollinated pistils. In particularly cool or overcast conditions a
week may be necessary, but the bag is removed at the earliest safe time to
ensure proper seed development without stray pollinations. As soon as the
bag is removed, the calyxes begin to swell with seed, indicating successful
fertilization. Seed parents then need good irrigation or development will be
retarded, resulting in small, immature, and nonviable seeds. Seeds develop
fastest in
warm weather and take usually from two to four weeks to mature
completely. In cold weather seeds may take up to two months to mature. If
seeds get wet in fall rains, they may sprout. Seeds are removed when the
calyx begins to dry up and the dark shiny perianth (seed coat) can be seen
protruding from the drying calyx. Seeds are labeled and stored in a cool,
dark, dry place, This is the method employed by breeders to create seeds of
known parentage used to study and improve Cannabis genetics.
Seed Selection
Nearly every cultivated Cannabis plant, no matter what its future, began
as a germinating seed; and nearly all Cannabis cultivators, no matter what
their intention, start with seeds that are gifts from a fellow cultivator or
extracted from imported shipments of marijuana. Very little true control can
be exercised in seed selection unless the cultivator travels to select
growing plants with favorable characteristics and personally pollinate them.
This is not possible for most cultivators or researchers and they usually
rely on imported seeds. These seeds are of unknown parentage, the product of
natural selection or of breeding by the original farmer, Certain basic
problems affect the genetic purity and predictability of collected seed.
1 - If a Cannabis sample is heavily seeded, then the majority of
the male plants were allowed to mature and release pollen, Since
Cannabis is wind-pollinated, many pollen parents (including early
and late maturing staminate and hermaphrodite plants) will
contribute to the seeds in any batch of pistillate flowers. If the
seeds are all taken from one flower cluster with favorable
characteristics, then at least the pistillate or seed parent is the
same for all those seeds, though the pollen may have come from many
different parents. This creates great diversity in offspring.
2 - In very lightly seeded or nearly sinsemilla Cannabis,
pollination has largely been prevented by the removal of staminate
parents prior to the release of pollen. The few seeds that do form
often result from pollen from hermaphrodite plants that went
undetected by the farmer, or by random wind-borne pollen from wild
plants or a nearby field. Hermaphrodite parents often produce
hermaphrodite offspring and this may not be desirable.
3 - Most domestic Cannabis strains are random hybrids. This is
the result of limited selection of pollen parents, impure breeding
conditions, and lack of adequate space to isolate pollen parents
from the remainder of the crop.
When selecting seeds, the propagator will frequently look for seed plants
that have been carefully bred locally by another propagator. Even if they
are hybrids there is a better chance of success than with imported seeds,
pro vided certain guidelines are followed:
1 - The dried seeded flower clusters are free of staminate
flowers that might have caused hermaphrodite pollinations.
2 - The flowering clusters are tested for desirable traits and
seeds selected from the best.
3 - Healthy, robust seeds are selected. Large, dark seeds are
best; smaller, paler seeds are avoided since these are usually less
mature and less viable.
4 - If accurate information is not available about the pollen
parent, then selection proceeds on common sense and luck. Mature
seeds with dried calyxes in the basal portions of the floral
clusters along the main stems occur in the earliest pistillate
flowers to appear and must have been pollinated by early-maturing
pollen parents. These seeds have a high chance of producing
early-maturing offspring. By contrast, mature seeds selected from
the tips of floral clusters, often surrounded by immature seeds, are
formed in later-appearing pistillate flowers. These flowers were
likely pollinated by later-maturing staminate or hermaphrodite
pollen parents, and their seeds should mature later and have a
greater chance of producing hermaphrodite off spring. The pollen
parent also exerts some influence on the appearance of the resulting
seed. If seeds are collected from the same part of a flower cluster
and selected for similar size, shape, color, and perianth patterns,
then it is more likely that the pollinations represent fewer
different gene pools and will produce more uniform offspring.
5 - Seeds are collected from strains that best suit the locality;
these usually come from similar climates and latitudes. Seed
selection for specific traits is discussed in detail in Chapter III.
6 - Pure strain seeds are selected from crosses between parents
of the same origin.
7 - Hybrid seeds are selected from crosses between pure strain
parents of different origins.
8 - Seeds from hybrid plants, or seeds resulting from pollination
by hybrid plants, are avoided, since these will not reliably
reproduce the phenotype of either parent.
Seed stocks are graded by the amount of control exerted by the collector
in selecting the parents. Grade #1 - Seed parent and pollen parent are known
and there is absolutely no possibility that the seeds resulted from pollen
contamination.
Grade #2 - Seed parent is known but several known staminate or
hermaphrodite pollen parents are involved. Grade #3 - Pistillate parent is
known and pollen parents are unknown.
Grade #4 - Neither parent is known, but the seeds are collected from one
floral cluster, so the pistillate seed parent age traits may be
characterized.
Grade #5 - Parentage is unknown but origin is certain, such as seeds
collected from the bottom of a bag of imported Cannabis.
Grade #6 - Parentage and origin are unknown.
Asexual Propagation
Asexual propagation (cloning) allows the preservation of genotype because
only normal cell division (mitosis) occurs during growth and regeneration.
The vegetative (non-reproductive) tissue of Cannabis has 10 pairs of
chromosomes in the nucleus of each cell. This is known as the diploid (2n)
condition where 2n = 20 chromosomes. During mitosis every chromosome pair
replicates and one of the two identical sets of chromosome pairs migrates to
each daughter cell, which now has a genotype identical to the mother cell.
Consequently, every vegetative cell in a Cannabis plant has the same
genotype and a plant resulting from asexual propagation will have the same
genotype as the mother plant and will, for all practical purposes, develop
identically under the same environmental conditions.
In Cannabis, mitosis takes place in the shoot apex (meristem), root tip
meristems, and the meristematic cambium layer of the stalk. A propagator
makes use of these meristematic areas to produce clones that will grow and
be multiplied. Asexual propagation techniques such as cuttage, layerage, and
division of roots can ensure identical populations as large as the growth
and development of the parental material will permit. Clones can be produced
from even a single cell, because every cell of the plant possesses the
genetic information necessary to regenerate a complete plant.
Asexual propagation produces clones which perpetuate the unique
characteristics of the parent plant. Because of the heterozygous nature of
Cannabis, valuable traits may be lost by sexual propagation that can be
preserved and multiplied by cloning. Propagation of nearly identical
populations of all-pistillate, fast growing, evenly maturing Cannabis is
made possible through cloning. Any agricultural or environmental influences
will affect all the members of that clone equally.
The concept of clone does not mean that all members of the clone will
necessarily appear identical in all characteristics. The phenotype that we
observe in an individual is influenced by its surroundings. Therefore,
members of the clone will develop differently under varying environmental
conditions. These influences do not affect genotype and therefore are not
permanent. Cloning theoretically can pre serve a genotype forever. Vigor may
slowly decline due to poor selection of clone material or the constant
pressure of disease or environmental stress, but this trend will re verse if
the pressures are removed. Shifts in genetic composition occasionally occur
during selection for vigorous growth. However, if parental strains are
maintained by in frequent cloning this is less likely. Only mutation of a
gene in a vegetative cell that then divides and passes on the mutated gene
will permanently affect the genotype of the clone. If this mutated portion
is cloned or reproduced sexually, the mutant genotype will be further
replicated. Mutations in clones usually affect dominance relations and are
therefore noticed immediately. Mutations may be induced artificially (but
without much predictability) by treating meristematic regions with X-rays,
colchicine, or other mutagens.
The genetic uniformity provided by clones offers a control for
experiments designed to quantify the subtle effects of environment and
cultural techniques. These subtleties are usually obscured by the extreme
diversity resulting from sexual propagation. However, clonal uniformity can
also invite serious problems. If a population of clones is subjected to
sudden environmental stress, pests, or disease for which it has no defense,
every member of the clone is sure to be affected and the entire population
may be lost. Since no genetic diversity is found within the clone, no
adaptation to new stresses can occur through recombination of genes as in a
sexually propagated population.
In propagation by cuttage or layerage it is only necessary for a new root
system to form, since the meristematic shoot apex comes directly from the
parental plant. Many stem cells, even in mature plants, have the capability
of producing adventitious roots. In fact, every vegetative cell in the plant
contains the genetic information needed for an entire plant. Adventitious
roots appear spontaneously from stems and old roots as opposed to systemic
roots which appear along the developing root system originating in the
embryo. In humid conditions (as in the tropics or a green house)
adventitious roots occur naturally along the main stalk near the ground and
along limbs where they droop and touch the ground.
Rooting
A knowledge of the internal structure of the stem is helpful in
understanding the origin of adventitious roots.
The development of adventitious roots can be broken down into three
stages: (1) the initiation of meristematic cells located just outside and
between the vascular bundles (the root initials), (2) the differentiation of
these meristematic cells into root primordia, and (3) the emergence and
growth of new roots by rupturing old stem tissue and establishing vascular
connections with the shoot.
As the root initials divide, the groups of cells take on the appearance
of a small root tip. A vascular system forms with the adjacent vascular
bundles and the root continues to grow outward through the cortex until the
tip emerges from the epidermis of the stem. Initiation of root growth
usually begins within a week and young roots appear within four weeks. Often
an irregular mass of white cells, termed callus tissue, will form on the
surface of the stem adjacent to the areas of root initiation. This tissue
has no influence on root formation. However, it is a form of regenerative
tissue and is a sign that conditions are favorable for root initiation.
The physiological basis for root initiation is well understood and allows
many advantageous modifications of rooting systems. Natural plant growth
substances such as auxins, cytokinins, and gibberellins are certainly
responsible for the control of root initiation and the rate of root
formation. Auxins are considered the most influential. Auxins and other
growth substances are involved in the control of virtually all plant
processes: stem growth, root formation, lateral bud inhibition, floral
maturation, fruit development, and determination of sex. Great care is
exercised in application of artificial growth substances so that detrimental
conflicting reactions in addition to rooting do not occur. Auxins seem to
affect most related plant species in the same way, but the mechanism of this
action is not yet fully understood.
Many synthetic compounds have been shown to have auxin activity and are
commercially available, such as napthaleneacetic acid (NAA), indolebutyric
acid (IBA), and 2,4-dichlorophenoxyacetic acid (2,4 DPA), but only
indoleacetic acid has been isolated from plants. Naturally occurring auxin
is formed mainly in the apical shoot men stem and young leaves. It moves
downward after its formation at the growing shoot tip, but massive
concentrations of auxins in rooting solutions will force travel up the
vascular tissue. Knowledge of the physiology of auxins has led to practical
applications in rooting cuttings. It was shown originally by Went and later
by Thimann and Went that auxins promote adventitious root formation in stem
cuttings. Since application of natural or synthetic auxin seems to stimulate
adventitious root formation in many plants, it is assumed that auxin levels
are associated with the formation of root initials. Further research by
Warmke and Warmke (1950) suggested that the levels of auxin may determine
whether adventitious roots or shoots are formed, with high auxin levels
promoting root growth and low levels favoring shoots.
Cytokinins are chemical compounds that stimulate cell growth. In stem
cuttings, cytokinins suppress root growth and stimulate bud growth. This is
the opposite of the reaction caused by auxins, suggesting that a natural
balance of the two may be responsible for regulating nor mal plant growth.
Skoog discusses the use of solutions of equal concentrations of auxins and
cytokinins to pro mote the growth of undifferentiated callus tissues. This
may provide a handy source of undifferentiated material for cellular
cloning.
Although Cannabis cuttings and layers root easily, variations in
rootability exist and old stems may resist rooting. Selection of rooting
material is highly important. Young, firm, vegetative shoots, 3 to 7
millimeters (1/8 to ¼ inch) in diameter, root most easily. Weak, unhealthy
plants are avoided, along with large woody branches and reproductive
tissues, since these are slower to root. Stems of high carbohydrate content
root most easily. Firmness is a sign of high carbohydrate levels in stems
but may be con fused with older woody tissue. An accurate method of
determining the carbohydrate content of cuttings is the iodine starch test.
The freshly cut ends of a bundle of cuttings are immersed in a weak solution
of iodine in potassium iodide. Cuttings containing the highest starch
content stain the darkest; the samples are rinsed and sorted accordingly.
High nitrogen content cuttings seem to root more poorly than cuttings with
medium to low nitrogen content. Therefore, young, rapidly-growing stems of
high nitrogen and low carbohydrate content root less well than slightly
older cuttings. For rooting, sections are selected that have ceased
elongating and are beginning radial growth. Staminate plants have higher
average levels of carbohydrates than pistillate plants, while pistillate
plants exhibit higher nitrogen levels. It is unknown whether sex influences
rooting, but cuttings from vegetative tissue are taken just after sex
determination while stems are still young. For rooting cloning stock or
parental plants, the favorable balance (low nitrogen-to-high carbohydrate)
is achieved in several ways:
1 - Reduction of the nitrogen supply will slow shoot growth and
allow time for carbohydrates to accumulate. This can be accomplished
by leaching (rinsing the soil with large amounts of fresh water),
withholding nitrogenous fertilizer, and allowing stock plants to
grow in full sun light. Crowding of roots reduces excessive
vegetative growth and allows for carbohydrate accumulation.
2 - Portions of the plant that are most likely to root are
selected. Lower branches that have ceased lateral growth and begun
to accumulate starch are the best. The carbohydrate-to-nitrogen
ratio rises as you move away from the tip of the limb, so cuttings
are not made too short.
3 - Etiolation is the growth of stem tissue in total darkness to
increase the possibility of root initiation. Starch levels drop,
strengthening tissues and fibers begin to soften, cell wall
thickness decreases, vascular tissue is diminished, auxin levels
rise, and undifferentiated tissue begins to form. These conditions
are very conducive to the initiation of root growth. If the light
cycle can be con trolled, whole plants can be subjected to
etiolation, but usually single limbs are selected for cloning and
wrapped for several inches just above the area where the cutting
will be taken. This is done two weeks prior to rooting. The
etiolated end may then be unwrapped and inserted into the rooting
medium. Various methods of layers and cuttings rooted below soil
level rely in part on the effects of etiolation.
4 - Girdling a stem by cutting the phloem with a knife or
crushing it with a twisted wire may block the downward mobility of
carbohydrates and auxin and rooting cofactors, raising the
concentration of these valuable components of root initiation above
the girdle.
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 a—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
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 don’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 florist’s tape or electrician’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-‘ 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.
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