Advancements and Research In The Field Of Cannabis Male Selection
Posted by zamalito at BBayhttp://www.breedbay.co.uk/forums/breeders-library/201338388-breeders-choice-breeding-male-selection.html

By The Researchers At Breeders Choice Organization (BCO)


We decided to create our company, Breeders Choice, to address several problems which we feel have become endemic in the Cannabis seed industry. At the forefront of these problems is that many breeding houses are actually first and foremost commercial growers. When it comes to female selection, this isn't really a problem. One of the goals of the commercial grower is to find successful and marketable female plants, and commercial growers can be very good at finding desirable females. Male selection, however, is quite different, and by no means compatible with commercial growing.

We view breeding of Cannabis as two parts art, with one part science, and one part spirituality. As with any art form, a truly great artist must be creative, have a sound technique, confidence in their abilities, a sense of aesthetics, and new innovative ideas. These are also the qualities that make a great breeder. The scientific aspect, I liken to musical theory. Science provides us with a language to communicate our ideas with our peers, a method to prove what techniques are effective, and on occasion, a muse or source of new ideas. The spiritual aspect helps put us in touch with the plants we work with. Cannabis is interesting, in that when we consume the plant, it speaks with us. Some plants are great teachers, and some plants are bland without much to say.

However, this concept that breeding is an art, has lead many to move in a destructive path away from quantifying traits, and studying the complex relationships which traits have to each other across individuals, and populations, and from a single generation to multiple generations. Each of these divisions are sentient beings within themself. The individual is the culmination of the many generations before it, and functions to pass its genetic material successfully. The population is just as much alive and dynamic as the individual, and is just as much a result of its DNA as the individual. The goal of the population is to create balance, allowing the most successful individuals to be the most successful but simultaneously allow for diversity to be maintained allowing for adaptation to new and changing ecosystems. While the individual is just a tool of the population that in the case of cannabis typically only survives through one reproductive cycle. The population survives forever, unless every individual fails to pass its genetic material on to another year. Over the many millions of years before the arrival of man, populations have evolved ways to protect future adaptability while undergoing thriving conditions and, lacking conditions/bottlenecks both of which can limit diversity without systems in place to protect it.


Many of us who were growing the staples of the Dutch seed industry, Skunk #1, Northern Lights, and Haze, in the 1980's, before and shortly after their arrival in Holland, remember remarkable plants. Sadly, these lines have been declining ever since, and most versions available are only a shadow of their pre-Dutch ancestors. The improvements and declines in the desirability of these lines are the most accurate barometer of the selection abilities of the seed industry, and its capability of line improvement. Having had more than a few glimpses into the selection techniques of most of the well known seed companies, we have come to the conclusion that the single greatest cause of this decline is poor male selection.

I'm sure many of you are wondering, how can this be an accurate barometer of the industry's abilities, when the potency of the seeds in circulation are constantly improving? Well, the answer to this lies in heterosis/hybrid vigor, and the constant introduction of new genes from outside sources. This isnt line improvement, and definitely isn't how you create new and unique breeding tools. Without someone, somewhere exercising proper selection, eventually the source of new and unique breeding tools will run dry, and the quality will begin to decline across the board.

The single most important thing any breeder whose goal is to produce desirable varieties of sinsemilla can do is to familiarize themselves with males. The pollen donor contribute heavily to the floral traits of their female offspring. However, males give more subtle clues about what they pass on to their female offspring. This being the case, every breeder should spend not only more time studying their male plants, but also spend exponentially more time finding good males, as opposed to females.

Even in the best natural environments, or sweet spots, which encourage higher potency, without constant selection, cannabis always has a tendency to revert to low potency. It may vary based on the original germplasm and the climate the seedlines are acclimating to, but an unselected seedline will almost without fail return to low potency, frequently below 4% THC, but sometimes a little higher depending on the original stock and the climate. Low potency is the default position of the species, with regards to potency.

Different Techniques For Different Projects

The first consideration one needs to make when planning a breeding project, is to define exactly what type of breeding project they are trying to proceed with.

Surprising to most people, conservation efforts, where one is trying to keep in tact, as much as possible, heirloom or landrace lines, tend to be the most technically difficult. It is the nature of genetics to be ing a constant state of flux, adapting to a particular environment. The nature of preservation breeding is to temporarily halt this state of flux, and this requires large populations of two thousand plants (1000 of each sex) minimum, fully completing their reproductive stage. You have to remember that passing every gene on to the following generation isn't enough. A population isnt just a set of genes, but is also how frequently these genes occur within the population. This is what makes the population alive and in a state of flux, and determines how the population responds to natural and unnatural selective pressure.

Hybridization tends to require the fewest parents, but as always it is best to make selections from as large of a population as possible, to find individuals with high specific combining abilities (SCA). All too often, the paint by numbers philosophy of breeding is applied where the breeder is more interested in crossing names, than actually applying the art of selection and the analysis or testing of combining abilities. To borrow a term from physics, individuals are either "in phase" with each other or "out of phase". Similar to a meeting of two electromagnetic waves, when individuals are "in phase", they amplify each others strengths and compliment each other. When out of phase, their strengths conflict, or their weaknesses amplify.

Phenotype fixing is the more common route used to create an inbred line (IL). Phenotype fixing is the type of breeding where the goal is to create a line which reproduces a particular phenotype as frequently as possible. The lines which try to put an elite cutting into seed form, as well as the work of DJshort belong in this category. Frequently, these projects involve one parent of each sex per generation and involve multiple backcrosses to an individual which reflects an ideal. They are rarely sustainable as populations, either because of extensive bottlenecking, or because of a high degree of heterosis causing the desirable phenotype to be fixed for only one generation. The problems with this technique are mainly because of the backcrossing and 2 parent per generation breeding style. The unnecessary removal of alleles which have no negative effect on desirability, brought by the use of small numbers of parents results in a lack of vigor and the

Population conditioning is a term we coined to refer to a type of breeding we try to focus on. The goal of this type of breeding isn't necessarily to make every plant of an identical phenotype, but to produce noticeable sustainable improvements with every generation, only removing traits which have a negative impact on the desirability of the line as a whole, and only selecting for traits which have a positive impact. The goal of our work is it to create genuine heirlooms and breeding tools which are unique, yet very desirable on their own. When applied in a hybrid breeding program, the conditioned lines create true heterosis, from being populations which never met before.

Our approach to population conditioning goes like this. With 1 X 1 incrossing the goal focus is to do the vast majority of the work in the f2 and f3 generations, with every other generation requiring a less substantial population to make selections from. With population conditioning it is kind of the opposite. In the f1 and f2, you can get away with using fairly small populations and open pollinating them, and as the generation numbers go up, so does the population size. The goal here is to Gradually eliminate undesirable traits, while not only keeping the desirable genes, but also genes which have no noticeable bearing on phenotype. These genes which have no noticeable bearing on phenotype, might not be immediately noticeable, but provide most of the combining potential when used as a breeding tool. After the population has been conditioned into a sustainable heirloom quality population (typically somewhere between f8 and f13, remember since multiple parents are being used, the population requires more generations to stabilize), we then begin to start working on splitting up the line into fixing particular phenotypes. These fixed phenotype lines do lack as much sustainability as the conditioned population line, but they do have breeding potential. Also, due to the undesirable traits being removed, the fixing of the phenotype can be done fairly rapidly, and with more potential of overall desirability.


What distinguishes the good projects of this sort from the more lacking landrace based IL's is the use of heirlooms which meet the quality standards of the old legends, as well as a very strict and proven male selection technique. It is one thing to grow out 20 seeds from an unimpressive line you were told were colombian gold bagseed, that flower in twelve weeks and cross it to an Afghani #1. It is a whole different story to select a 40 week flowering Colombian heirloom collected in Colombia that produces a truly legendary psychedelic high and cross it to well selected Mazar heirloom from the region of origin. In this instance, a new legend can be produced with unique qualities of its own.

Of course, we work with what we can get, and not everyone has access to the legends of old. We hope to change this. Now, even the best landrace based breeding materials often need some work to be consistently legendary, and consistent performing breeding tools. This is yet another use for the population conditioning approach, but judgement must be exercised so as not to remove to much of the diversity or desirability of the line before breeding with it. Also, any landrace or heirloom line must be preserved using as large of an open pollination as possible before trying to improve the line with either incrossing or outcrossing.

Numbers Games:

Cannabis is a wind pollinated species, and as such, even the lines which appear to be heavily consistent, there is still significant genetic diversity. This means that no line is safe from improvement brought by good selective breeding, or ruin, brought by poor selection. As a whole, the species tends not to be compatible with small packets of ten seeds, and little good can come from populations this small. The high cost of seeds has caused us to nurse every single seed, having a detrimental effect on the ability of seeds to undergo adversity during germination.

Small populations are a relatively new thing to the cannabis species. All of the heirloom strains were grown field scale, and many parents were selected from very large populations. Originally in Thailand, there were farmers who produced seed for the rest of the community. This was the time of the great Thai herbs, which are now on their last limb. During this period, hermaphrodites were not anywhere near as common in Thai populations as they are now. In a kilo, you might find 2-3 seeds in the high quality Thai sticks. When the demand for Thai cannabis grew, the growers who provided seed for their community, ceased to do so, and growers started to pick seeds out of their female crops to grow the next generation, unintentionally breeding for hermaphrodites.

The size of the population you grow, is in essence, the size of the breeder's pallet. A limited pallet not only limits the improvements that can be made, but also limits what can be done creatively/artistically. On the other hand, when a breeder has no effective selection technique or is incapable of recognizing the differences in the plants, the size of the pallet doesn't really matter.

The number of parents selected from the population to produce the following generation is also very important. The more parents selected, the greater the genetic diversity passed on to the following generation. It is important to note that genetic diversity isn't always a good thing, nor is it always a bad thing, it depends on what you are doing. Also, the greater the number of parents needed to accomplish the desired effect, the greater the size of the population that must be grown to still select plants which meet your standards for desirability. We on a rare occasion will select one male and one female for one or two generations between the p1 and f3. Cannabis being a wind pollinated species, populations don't seem to respond as well when this technique is applied for two generations in a row. We only apply this method when either the individuals that meet our standards are very difficult to find, or there is simply too much diversity, such as the f2 from a 3-way hybrid.

Since a greater number of parental offspring, passes on more genetic diversity to the resultant offspring, the risk of deselecting genes unnecessarily which either have a positive or limited effect on desirability as well as the risk of recessive genes which cause deformities and negative traits is greatly reduced. By removing genes which have little to no immediately perceived effect on consistency or desirability, the breeder may inadvertently be reducing vigor and resistance to a pathogen or pest which the line has not yet been exposed. The alignment of recessive alleles from unnecessary bottlenecking and closely related gene pool is the primary cause of weird mutant looking plants, and deformities. Although it does increase the number of generations required to achieve a consistent seedline, a higher number of parents limits the liability of inbreeding, solidifies advancements, and actually hastens the fixing of rare traits when done properly.


Tools Of The Trade

The first tool that should be in the arsenal of any breeder is a good digital camera, capable of taking high quality macros, with minimal distance between the lens and the object being photographed. The camera allows for close up examination of gland types, pattern and structure, and also allows for cataloging of traits, and specimens as well as direct side by side comparison on your computer.

After a digital camera, and for many of the same reasons is a good binocular microscope, of the type used for making dissections, and capable of taking digital images. This will allow in depth study of both glandular and nonglandular trichomes, as well as tissue samples.

A refractometer/brix meter is a tool used by growers of wine grapes. These are used to measure the contents of the plant juices, primarily sugars. This isn't an extremely important tool in cannabis breeding, as much as it facilitates cultivation, but does allow one a glimpse of the metabolic processes in the plant. Any clues that one can associate with desirable phenotypes early on is useful, as it helps to thin plants at an earlier stage, thus increasing the size of the seed population, the breeder can start.

Thin Layer Chromatography tests (TLC) such as the Cannalyze (TM) are not very effective in quantitative analysis, but when the samples are taken using very precisely measured samples using milligram accurate scales for measurement can be effective for comparative potency. Although they might not achieve a precise ratio of THC to plant matter, all a breeder really needs to know is how potent they are in relation to one another. Where TLC really comes in handy is in the use of breeding medicinal cultivars with limited recreational potential, because of its ability to recognize the presence of various cannabinoids. Unfortunately these tests aren't sensitive enough to measure accessory cannabinoids in the male flowers so using them for comparative floral potency between males isn't the best idea. Testing male leaf material and female flowers for high CBD levels is where this method really shines. Unfortunately CBD has little use in recreational cannabis, due to it blocking the effects of THC. All of the commercially available varieties from the many cannabis seed companies are very low in CBD levels, despite what they may advertise.

Antibody testing methods such as ELISA where a sample is applied to a microplate coated with antibodies which specifically detect THC, are extremely accurate and sensitive. Unfortunately, to get an accurate percentage of THC concentrations several different samples of known concentration must be used and put into an inexpensive or free version of data plotting software. If no samples of confirmed purity are known, then this method can only be used for comparative results. Comparative results, are however, good enough for selecting the samples which test the highest among a population.

Having a method to gauge THC or other cannabinoid levels is great, but cannot fully replace an accurate scale, and some rolling papers. THC and other cannabinoid concentrations do not directly translate into perceived potency. For this reason, there is no replacing a subjective bioassay. Many compounds from diverse groups such as sesquiterpenes, monoterpenes and other VOC substances such as, aromatic compounds, non-terpene hydrocarbons, esters and organic sulfides, as well as their combustion by-products. Smoking of males is somewhat controversial among breeders. Some breeders say that since males have a different ratio of glandular types, the high does not directly translate into the female offspring, and this is true to some extent. Others say that males are simply too weak to get a quantifiable effect. We do have ceilingless, soaring, and psychedelic males that pass these traits to their offspring quite consistently. Many of our selected males, would make my smoking tray before many types of hashish. They hardly ever taste as good and are never as potent after one or two hits, but their ceilingless, soaring effect is unquestionable, and this is what you look for when smoking males. Another thing you look for when smoking males is a substantial increase between the shoots and floral leaves over the fan leaves. Plants which have a more similar potency across the whole plant, tend to not be as potent overall.

Creating Diversity Through Inbreeding

At first this sounds like an alien concept. After all, how can you create diversity in a line without adding new genes? Well, there are different types of diversity. There is phenotypic diversity, allelic frequency, and allelic diversity. For example, true f1 crosses are known to be very consistent and rarely produce a huge diversity of phenotypes, but should be genetically diverse. The f2 of the same cross should have less genetic diversity than the f1, but due to the appearance of the recessive phenotypes, more genetic diversity can be physically expressed, showing phenotypes all the way from one parental line to the other and everywhere in between, as well as new traits.

A line can have 50 different alleles (genes) for a given loci giving the loci a diverse array of genes, but if 49 of those only occur in 1% of the population, then the particular loci will be fairly homogenous. This is the difference between allelic frequency and diversity, and shows how they effect homogeneity. If the breeder increases the frequency of these rare alleles, the pallet of the breeder and the diversity commonly expressed in the line also broadens.

By selecting parents from the fringes of the population and rare phenotypes, diversity and vigor can be created where none existed before. This is where population conditioning comes into effect. In 1X 1 incrosses, rare phenotypes rarely pass on their traits, but when 5 parents of each sex are used which all display the same rare pheno, the trait tends to be fixed more heavily into the resultant offspring.

The Nuts And Bolts Of Male Selection


The first thing anyone should do when working with a new potential line in a breeding project is to familiarize yourself with the strain. Look at how the male phenotypes show similarity to female phenotypes and which phenotypes most consistently produce the traits which you seek. Detail is key, look at whether plants male plants produce glandular trichomes on the unopened flowers and the floral leaves. What variance is there in the effects of smoked males and trichome production in reproductive regions, and how is this variance similar to the females?Most adequately potent varieties of cannabis produce a row of glands on the anthers, the banana shaped and colored portion of the male flower. Take particular note on the size, shape, frequency, and density of the anther glands.

The tricky thing about selection of both males and females is that unless you are working with a mapped genome, and have the tools to analyze the genetic code, you have to make your selections based on phenotype, as in the physical manifestation of genetic traits. However, what makes a good parent is its genes, not physical traits. There are many things that get in the way of a genotype's physical expression, and a female plant which might be the best plant you have ever come across, may very well be a mediocre breeding plant. Many facets of the environment which are seemingly innocuous can play a huge role in the expression of a plant's genes. I have seen many outdoor plants which produced some of the best sativa dominant flowers I've ever grown, but when grown indoors be completely bland and actually lean towards an indica. One reason for this is that genes compete for a limited quantity and type of resources. If any of those resources aren't present in the proper quantity, then some traits might fail to express completely, even if the plant was grown perfectly in a different environment.

It goes without saying, that breeding of wind pollinated plants traditionally grown field scale, and breeding of race horses are extremely different, but this brings to mind, a horse named Storm Cat. Storm Cat ranks among the highest paid athletes in the world, despite his racing record which is far from the stuff of legends. But, the reason this horse has made so much money for his owners is not because he is a great race horse, but because his children are. You see, Storm Cat is a stud horse, earning $500,000 per breeding session. His offspring have earned almost $100 million dollars on the race track. Great breeding plants are similar in this way. They might not be the greatest plants to smoke, although they are rarely terrible, but they have that magic in their genes which makes their offspring legends.

This is where the similarities start to end between plant and animal breeding. I see it all the time where breeders apply the philosophy of pedigree horse, pig, or dog breeding to cannabis, partially out of the hype of elite cuttings, and partially out of misapplied logic. With cannabis, as I have stated before, the population is a living organism just as much as the individual. The population evolves and adapts, and populations of the same species compete for niche space just as much as individuals do, and like a pack of African wild dogs (another animal analogy, I know) the functionality of the population effects its survival just as much as that of the individual. On the other hand, with pedigree animal breeding, it is almost always all about the individual. The mechanics of heredity between the two reflects this. Genetics have evolved what we call an "emergent behavior" which is most similar to a decision making process ingrained in the genetic code. For example, we have seen how plants which exhibit a phenotype, which is rare among it's own siblings, are selected for two successive generations of 1 X 1 incrossing tend not to fix this phenotype in any statistically significant manner . This is the reason why elite cuttings tend to make mediocre breeding parents, because they often stand out from their siblings. However, when you select twenty plants, all exhibiting the same phenotype, representing a disproportionate percentage of the population exhibiting this phenotype, for two generations, the trait is conditioned into a greater portion of the population.

Matricliny and patricliny, are perhaps the two most overlooked and misunderstood facets of cannabis breeding. I believe I have this concept pinned down, but like many facets of this species, the more I believe I have it pinned down, the less I believe I can predict it. That said, one of the most important things to observe when studying test crosses made from a specific line or parent, is to very closely observe which traits the plant tends to pass, with specific focus on the pigmentation traits, terpenes, and the psychoactive effects. These traits in particular are not effected by the natural laws of heritance, and will be proving and studying the mechanics of this more in the future.

An important part of breeding is to find clues in the phenotype which help determine both the combining abilities and the genotype of the plant being examined. For example, if one plant is very strong despite having little resin, and another has heavy production of resin but less potent, then combining them might possibly produce plants with lots of potent resin, thus being more potent than either parent. There are some traits which can be used to determine the genotype you seek but have little to do with the actual phenotype, and may even be less effected by the environment. Finding traits such as these that have been proven through testing are really crucial in developing a great male selection technique. This is the most secretive aspect of our proprietary selection technique, but I can give some examples to help illustrate exactly what I am talking about. For example, certain aspects of the leaf surface can give clues to the genotype regarding glandular trichome production even more so than the glandular trichome production itself, since glandular trichome production doesn't happen much on males, and on females doesn't occur until late and is greatly effected by environment. The leaf surface traits however, aren't effected much by environment, and are visible with a binocular microscope from the very earliest stages.

In Conclusion

Over the coming months, HHF and I will be expanding on this and other subjects here with a series of articles, and sharing as we learn. In summation, this article can be summed in a few sentences. First of all, observation is key, as breeders, it is crucial to pour over every nanometer of both individuals and populations, and how these change when environments and light cycles change. Second of all, use or invent any tool to possibly measure and quantify every trait you observe. Finally, "when in doubt, grow more out", never underestimate the importance of population size.

The Breeders Bible is another great resource on Male selection....GooeyBreeder has some more info thats Great as well..

Hey @Drfting07 can you share the link with the others Here?
I have the Book

I have GooeyBreeder permission to copy his threads and info post just aint gotten around to them