How do I create a true breeding strain?
I've been hearing a fair bit of confusion from many on how to create a true breeding strain and so I'm writing this page to try and help shed some light on the subject. There are a few situations where a plant breeder would want to create a true breeding strain (IBL) and a few ways of accomplishing the task. But understanding the subtle differences of the various techniques is not so easy.
This paper will attempt to give a basic understanding of what is actually happening with each technique and then apply what is learned to actual projetcs. As a friend worked overtime making sure I didn't forget, breeding is not a black and white subject and as a whole, it would be too complex to put on paper in an easily understood form. Therefore, I will create small fictional examples to reinforce various concepts and then we will take those examples and concepts and apply some reality to them.
Try not to get hung up on the erroneous assumptions used here such as flavour being monogenic, the assumption is simply used to make it easier to learn a certain concept.
Just What Is It That We Are Doing?
Before we dive in, maybe we should take the time to understand what we are trying to accomplish when we set out to create a true breeding strain. There are hundreds of possible phenotypic traits that we could observe within a cannabis population. Are we trying to make all of them the same and remove ALL variation? Not likely, the genetic code is just too complex to try. Plus, since phenotype (what we see) is 1/2 genotype + 1/2 environment, everytime the population was grown under new conditions, new heterozygous traits would be observed. Basically, all we are trying to create is an overall uniformity while not worrying about the minor individual variations. No different than a dog breed. You can look at a german shepard and recognise it as belonging to a discrete breed. But if you look closer at several german shepards all at the same time, you will find variations with each and every one of them. Some will be a little taller, some a little wider, some more agressive, some a little fatter, some darker, etc. But they would all fall within an acceptable range for the various traits. Generally speaking, this is what a plant breeder is trying to accomplish when creating a true breeding strain, or IBL.
However this isn't always the case. Sometimes a breeder will just concentrate on a specific trait, like say outdoor harvest date, or mite resistance. You could still have a population where some are 2' bushes and some 10' trees. In this case, you would say that the strain was true breeding for the particular trait, but you wouldn't consider it true breeding strain per se. In genetics, wording plays a big part in meaning and understanding. As does point of reference as my F1 vs F2 comparison page illustrates.
Ok, so we want to make a cannabis population fairly uniform over a few phenotypically important traits, like say flavour for instance. For simplicity sake, we'll just deal with the single trait flavour, it's complex enough. And although flavour is controlled by several gene pairs (polygenic), we'll make the simplistic assumption that it's controlled by a single gene pair (monogenic) for many of the models and examples in this paper. There are many flavours such as chocolate, vanilla, musky, skunky, blueberry, etc, but in this paper we'll just deal with two flavours, pine and pineapple. Either gene in the gene pair can code for either of the flavours. If both genes code for pineapple or both genes code for pine flavour, we say that the gene pair (and individual plant) is homozygous for flavour. If the one gene codes for pine and the other codes for pineapple, we say that the gene pair (and individual plant) is heterozyous with respect to flavour. The heterozygous individual can create gametes (pollen or ovules) that can code for either pine flavour or pineapple flavour, the homozygous individuals can only create gametes that code for one OR the other. A homozygous individual is considered true breeding and a heterozygous individual is not.
However, as the words imply, when we are creating a true breeding strain, we are looking at a population, not individuals. We are trying to make all the individuals in the population homozygous for a particular trait or group of traits. Lets say we have a population of 50 individual plants, and each plant has has a gene pair coding for flavour. That means that 100 flavour genes make up the flavour genepool (reality is much more complex). When trying to create a true breeding strain, we are in fact trying to make all 100 of those genes code for the same trait ( pineapple flavour in our case). The closer our population comes getting all 100 genes the same, the more homozygous or true breeding it becomes. We use the terminology gene frequency to measure and describe this concept, where gene frequency is simply the ratio or percentage of the population that actually contains a specific gene. The higher the gene frequency, the more true breeding the population is. A fixed trait is where the gene frequency of the trait reaches 100%.
And folks, this is the basic backbone of what breeding is all about, manipulating gene frequencies. It doesn't matter if your making IBL, F1s, F2s, selecting for this or selecting for that, all you are really doing is manipulating gene frequencies. Therefore, to ever really understand what is happening in any breeding project, the breeder must pay attention to gene frequencies and assess how his selective pressures and models are influencing them. They are his measure of success.
What are we trying to create a true breeding strain from?
This a good question. Sometimes a gardener will notice a sport or unique individual in an F2 population, like say it has pineapple flavour when the rest have pine flavour. For one reason or another he decides he wants to preserve this new trait or combination of traits from that single individual. For the sake of ease of comprehension, we tend to call this special unique individual the P1 mom. He could start by selfing the individual OR breeding that individual with another and create what can be described as F1 offspring. If the F1 route was chosen, then breeders can diverge down two new paths. Some breeders will take the progeny of the F1 crossing and breed it back to the P1 mom, and then repeat for a couple more generations. This is referred to as backcrossing or cubing by cannabis breeders. Another common strategy is to make F2 progeny from the F1 population and then look for individuals that match the P1 mom. They would repeat the process for a few generations. We can call this filial or generational inbreeding since the parents from each cross belong to the same generation.
In another situation, sometimes a farmer will notice a few individuals in his fields that stand out from the crowd in a possitive manner. Like say the are resistant to a problem pest like powdery mildew. In this case, he will collect the best of the individuals and his starting population will contain several similar individuals and not a unique single individual as in the previous example. He would skip the hybridizing step (making the F1s) and go straight to the generational inbreeding step. Links to pages going into detail of each of these basic techniques and their impact on influencing gene frequencies are at:
A) Selfing the individual
B) Backcrossing and Cubing
C) Filial or Generational Inbreeding from an individual
D) Filial or Generational Inbreeding from a group
Applying the Pressure
Another excellent method to influence gene frequencies is to apply selective pressure. The idea here is to select only individuals that carry the desireable genes, and discard the rest.
A) Principles of selection
B) Progeny tests