breeding-crossing Pepper crossing question

[armyofda12mnkeys]

Hey all,

I wanted to get into pepper crossing this year. I kinda did last year but didn't emasculate the receiving flowers at all, so left it up to chance to see if they would pollinate by brushing the flower I wanted to use as my 'male' and then putting a very small seed ziplock plastic around it that also contained the male flower touching the pistol just in case my initial transfer didn't work. I do see somewhere between 25% to 50% of my crosses are much different looking so it was somewhat a success. I'll do the real steps (aka emasculating) for new crosses i want to make to get that up to 100% this year . But anyway I had a question.

For the crossing guide on https://www.fatalii.net/growing_chile_peppers/breeding (chart below), they mention: "PF = F1 hybrids partially fertile", what does that mean?
The seeds of the cross I got last year that I am growing this year, aka my first year of the cross (aka F1), the seeds of those may or may not be fertile?
So if they grew out this spring and became plants this year, then Im good since that proves that particular F1 seed was Fertile? and every single plant of next year's generation will be viable/fertile and so on and so on (just have to stabilize the variation/genetics i want)?

or does that mean this years upcoming F1's seeds (which will become next years F2 plant) will only be partially fertile? So some of those seeds i collect this year may or may not actually grow next year?

Sorry if that was confusing, but hopefully someone can answer .

NG = F1 hybrids germinate normally
EC = F1 hybrids raised by embryo culture
IV = fruits/seeds set, but F1 seeds inviable
PF = F1 hybrids partially fertile
HF = F1 hybrids highly fertile

 

[armyofda12mnkeys]

Hey AOT12M. Sounds like you'll have an interesting season ahead working with your hybrids.

I'm still not 100% on the pungency genetics, but my understanding of this is that capsaicinoid production is generally controlled by a single dominant C (Pun) gene, but capsaicinoid content is a quantitatively inherited trait involving multiple loci over multiple chromosomes.

I've read that the recessive (c or pun-1) non-pungent trait is found (naturally) only in annuum. For chinese, there is a less common mutation at a different locus, the recessive lov (loss of vesicle) that results in non-pungency.

So, if you're dealing with either, but not both, of c/pun-1 or lov, the presence of pungency would be a single-locus dominant trait, F1's would be phenotypically pungent, and a monohybrid punnett square model would apply.

But, if you're dealing with parents where each is complimentarily dominant at one pungency locus and recessive at the other (an aaBB x AA/bb scenario), a dihybrid punnett square model should apply with complementation figuring into the model. Complementation would indicate 100% pungency at F1 as well as pungency in any event of aaBB or aaBa type result, phenotypically masking a stabilized recessive "non-pungent" pairing at only one of the loci. By comparison, a non-complementary dihybrid model would allow you to see either of the two recessive pairings phenotypically, making the selection and recombination process easier.

Hey CaneDog, Thanks for that info.
I can kinda follow along (especially after watching this video on DiHybrid Punnet Squares to re-catchup on how to do those)...

But I guess I still have a question that if my cross comes from 2 Capsicum Chinenses... then that rules out the "recessive (c or pun-1) non-pungent trait is found (naturally) only in annuum" peppers.
So does that mean I only need to do a 'regular' monohybrid punnett square with the two "the recessive lov" genes of Parent#1(Grenada Seasoning) and the probably two regular/dominant "C (Pun) genes" of Parent#2(Pérola Iaranja)? (I'm assuming for now P.I. isn't heterozygous if its a stable pepper people sell).
That would mean my above tables would be accurate which would be great , which would mean 25% of my F2's would have a pair of this recessive lov genes.

Now for my 2nd cross, they were from 2 'no-heat' Chineses peppers, so I'm guessing that it would have the recessive gene across the board in the children which would be very cool (the F1 was also no-heat, but i guess we'll see how F2 goes).

Lastly, getting back to the degree of capsaicinoid content being a quantitatively inherited trait, when pungency is expressed phenotypically there remain at least several other loci to consider as to the degree of pungency, which might range from barely perceptible to melts-your-face-off

This is kinda interesting ... trying to absorb this info ... so lets say above chart is true and 25% have the non-pungency trait (which i guess i would translate in english to the 'no/low-heat' trait) but the 'degree of heat' is another trait that can still influence the level of heat?
So could it be non-pungent, AND have a medium or high heat-level still?

I guess it would be good to know we are on the same page about the word "non-pungent", I assume that means a 'no or very low heat level' trait vs pretend a 'Habanero flavor' trait that things like Habanada/AjiDulce/TrinidadPerfume/GrenadaSeasoning have (I also want that 'flavor' attribute to make its way into the F2's ).

 

[CaneDog]

Hey CaneDog, Thanks for that info.
I can kinda follow along (especially after watching this video on DiHybrid Punnet Squares to re-catchup on how to do those)...

But I guess I still have a question that if my cross comes from 2 Capsicum Chinenses... then that rules out the "recessive (c or pun-1) non-pungent trait is found (naturally) only in annuum" peppers.
So does that mean I only need to do a 'regular' monohybrid punnett square with the two "the recessive lov" genes of Parent#1(Grenada Seasoning) and the probably two regular/dominant "C (Pun) genes" of Parent#2(Pérola Iaranja)? (I'm assuming for now P.I. isn't heterozygous if its a stable pepper people sell).
That would mean my above tables would be accurate which would be great , which would mean 25% of my F2's would have a pair of this recessive lov genes.

Now for my 2nd cross, they were from 2 'no-heat' Chineses peppers, so I'm guessing that it would have the recessive gene across the board in the children which would be very cool (the F1 was also no-heat, but i guess we'll see how F2 goes).


This is kinda interesting ... trying to absorb this info ... so lets say above chart is true and 25% have the non-pungency trait (which i guess i would translate in english to the 'no/low-heat' trait) but the 'degree of heat' is another trait that can still influence the level of heat?
So could it be non-pungent, AND have a medium or high heat-level still?

I guess it would be good to know we are on the same page about the word "non-pungent", I assume that means a 'no or very low heat level' trait vs pretend a 'Habanero flavor' trait that things like Habanada/AjiDulce/TrinidadPerfume/GrenadaSeasoning have (I also want that 'flavor' attribute to make its way into the F2's ).

Yep. I think you're probably dealing with lov, so yeah.

I believe non-pungency to be an absolute. If there's pungency, it could range significantly. If there isn't pungency, I would expect the underlying genetic factors that affect degree of pungency to be present and these would then come into play if pungency genes were later introduced. So, if you hybridized a very hot pepper with a non-pungent pepper having high pungency supporting genetics you would get a hotter result than if you bred the hot pepper with a non-pungent pepper having low pungency supporting genetics.

This could come into play were you to cross a no-heat chinense with a very low heat chinense. If the no-heat chinense had high pungently supporting genetics they could come into play with the low-heat chinense's pungency contribution and result in a solid medium heat pepper from two parents, which appeared to both be no-heat.

 

[armyofda12mnkeys]

Thanks again for all this info @CaneDog !
I have maybe one last question... so this years F2 generation ill be seeing lots of variation.
I assume its fine of course if each individual plant self-pollinates, whether that be from:
Plant#1 Flower#1's male stamen to its own (Plant#1) Flower#1 female stigma
or
Plant#1 Flower#1's male stamen to a different flower on the same exact plant, so to (Plant#1) Flower#2 female stigma.
I assume both scenarios are fine, so I can cover a whole plant with a mesh (or individual branches with a large ziplock, or individual flowers with tea-bags or small seed ziplocks).

But I should not have this scenario i believe:
Plant#1 Flower#1's male stamen to Plant#2 Flower#2 female stigma.
as that Plant#2 probably came from a seed from different fruit pod on last year's F1 plant and hold different genetics which would make the process longer of stabilizing this years plants going forward, since it basically introduced another cross.

P.S. I gave away 2000 seeds (maybe some are more serious gardeners though) so hoping I get 500 plants to tested out to find my mini-heatless pepper .

 

[CaneDog]

Thanks again for all this info @CaneDog !
I have maybe one last question... so this years F2 generation ill be seeing lots of variation.
I assume its fine of course if each individual plant self-pollinates, whether that be from:
Plant#1 Flower#1's male stamen to its own (Plant#1) Flower#1 female stigma
or
Plant#1 Flower#1's male stamen to a different flower on the same exact plant, so to (Plant#1) Flower#2 female stigma.
I assume both scenarios are fine, so I can cover a whole plant with a mesh (or individual branches with a large ziplock, or individual flowers with tea-bags or small seed ziplocks).

But I should not have this scenario i believe:
Plant#1 Flower#1's male stamen to Plant#2 Flower#2 female stigma.
as that Plant#2 probably came from a seed from different fruit pod on last year's F1 plant and hold different genetics which would make the process longer of stabilizing this years plants going forward, since it basically introduced another cross.

P.S. I gave away 2000 seeds (maybe some are more serious gardeners though) so hoping I get 500 plants to tested out to find my mini-heatless pepper .

Yep. That's exactly how I understand it. In nature, the tendency of individual pepper plants to self-pollinate (occurring at the flower level based on the mechanics) is what causes segregation to occur so readily over time without intervention. You seem to have a clear understanding of this, but I'll reference this article that covers the basics - page 1 & page 2 (IIRC, page 2 addresses this specifically).

In a controlled population, enforcing self-pollination at plant level is sufficient, even though the mechanics of natural self-pollination essentially control it to the flower-specific level - i.e., without intervention most natural growing pepper plants will have a significantly high occurrence of self-pollination events at the flower level, with the remaining pollination events most likely distributed fairly evenly between pollination of a flower on the plant with a different flower on the same plant and a flower on the plant with a flower of an nearby plant.

Wow! Even a tenth of so many plants would provide excellent chances to see and select the best and the brightest from your combinations. Hopefully some really cool attributes will show!

 

[armyofda12mnkeys]

Yep. That's exactly how I understand it. In nature, the tendency of individual pepper plants to self-pollinate (occurring at the flower level based on the mechanics) is what causes segregation to occur so readily over time without intervention. You seem to have a clear understanding of this, but I'll reference this article that covers the basics - page 1 & page 2 (IIRC, page 2 addresses this specifically).

In a controlled population, enforcing self-pollination at plant level is sufficient, even though the mechanics of natural self-pollination essentially control it to the flower-specific level - i.e., without intervention most natural growing pepper plants will have a significantly high occurrence of self-pollination events at the flower level, with the remaining pollination events most likely distributed fairly evenly between pollination of a flower on the plant with a different flower on the same plant and a flower on the plant with a flower of an nearby plant.

Wow! Even a tenth of so many plants would provide excellent chances to see and select the best and the brightest from your combinations. Hopefully some really cool attributes will show!

Awesome, thanks for that info.
I have read those tomato genetics webpages a few times in the last month. Definitely very useful and really cool visual way to show whats going on genetically for those simple kinds of Dominant/recessive attributes .

 

[armyofda12mnkeys]

I believe non-pungency to be an absolute. If there's pungency, it could range significantly. If there isn't pungency, I would expect the underlying genetic factors that affect degree of pungency to be present and these would then come into play if pungency genes were later introduced. So, if you hybridized a very hot pepper with a non-pungent pepper having high pungency supporting genetics you would get a hotter result than if you bred the hot pepper with a non-pungent pepper having low pungency supporting genetics.

This could come into play were you to cross a no-heat chinense with a very low heat chinense. If the no-heat chinense had high pungently supporting genetics they could come into play with the low-heat chinense's pungency contribution and result in a solid medium heat pepper from two parents, which appeared to both be no-heat.

Hey @CaneDog , maybe my last question hopefully ... so many interesting questions pop up in my head , but think this is my last important one...
Would these peppers that I'm interested in re-creating with my hybrid ... AjiDulce/TrinidadPerfume/Habanada/GrenadaSeasoning...
Would you consider them:
'no-heat' peppers (which may or may not have low or high pungency supporting genetics ... we don't know since they would be hidden by the no-heat gene)
or
'has heat' peppers with a low pungency (aka very low heat-level the pungency genetics contribute to)?

I'm curious since one of the parents (Grenada Seasoning) is a low/no-heat pepper contributing to the children's genes and I'm curious what style of pepper that possibly is.

If you are not sure, is there any good pepper geneticist PHD to contact ?

 

[CaneDog]

Unfortunately, I don't know and I don't remember any papers I've read discussing this aspect of their genetics.

My first thought was they might be non-pungent varieties, because the similar with-heat peppers are considerably hot and I think it would be more likely they obtained non-pungent genetics in one fell swoop (a single locus mutation) rather than becoming less pungent over a longer period of time and selection (multiple loci).

On the other hand, I've heard people say they've tried some of these and detect low-to-no-heat, which suggests otherwise. So, perhaps they are pungent and were selected, with or without human intervention, for lower pungency over time. There certainly is reason people would select for less hot versions of similar peppers.

Perhaps you'll be the one to figure this out!

 

[armyofda12mnkeys]

I'll post some pics at some point later when the seedlings get a bit bigger, but I thought it was interesting that a small percentage%... pretend 3-5% out of 100 seedlings so far have purple stems and purple-tinted leaves (which I've experienced that usually means purple fruit via anthocyanin... Is that always the case that the fruit will have some purple as well ???).
Thought that was interesting as there was no purple genetics, as far as I know, in the parents.

 

[CaneDog]

In my experience, a mix or purple and green stems early on is a not all that uncommon within an otherwise "stabilized" population. Purple vs green leaves not so much. Sometimes darker leaf coloration is triggered by the light intensity a sprout is receiving and might go away as the sprout gets more established with sets of true leaves. I don't know that dark foliage always means purple tinted pods, but they do seem to correlate.

Looking forward to the pic's!

 

[armyofda12mnkeys]

Something I just realized reviewing my Youtube video, Pérola laranja fruit has a slight purple blush when unripe as seen here (although the plant/leaves itself is not purple);
So maybe the parents do have some purple genetics somewhere in there if this blush is considered anthocyanin expression?.

Btw when I googled Pérola laranja, I noticed the only other person really mentioning it was you ...

Do you think Pérola laranja is a frutenscens/chinense hybrid itself? Most of the stores selling it classify it as chinense.
You could have been saying maybe the other pepper is a hybrid and was just using P.I. as a comparison though.

 

[CaneDog]

I've always though of orange pearl just as a chinense and don't remember anything from growing it that would cause me to think differently - not that there couldn't be something. I was just noting that the peach frutescens I grew has an annular constriction indicative of chinense and speculating that perhaps it resulted from a cross with a chinense similar to the OP.

I got plenty of antho sun-blush on the orange pearl , too.

 

[PollenNut]

Hey all,

I wanted to get into pepper crossing this year. I kinda did last year but didn't emasculate the receiving flowers at all, so left it up to chance to see if they would pollinate by brushing the flower I wanted to use as my 'male' and then putting a very small seed ziplock plastic around it that also contained the male flower touching the pistol just in case my initial transfer didn't work. I do see somewhere between 25% to 50% of my crosses are much different looking so it was somewhat a success. I'll do the real steps (aka emasculating) for new crosses i want to make to get that up to 100% this year . But anyway I had a question.

For the crossing guide on https://www.fatalii.net/growing_chile_peppers/breeding (chart below), they mention: "PF = F1 hybrids partially fertile", what does that mean?
The seeds of the cross I got last year that I am growing this year, aka my first year of the cross (aka F1), the seeds of those may or may not be fertile?
So if they grew out this spring and became plants this year, then Im good since that proves that particular F1 seed was Fertile? and every single plant of next year's generation will be viable/fertile and so on and so on (just have to stabilize the variation/genetics i want)?

or does that mean this years upcoming F1's seeds (which will become next years F2 plant) will only be partially fertile? So some of those seeds i collect this year may or may not actually grow next year?

Sorry if that was confusing, but hopefully someone can answer .

NG = F1 hybrids germinate normally
EC = F1 hybrids raised by embryo culture
IV = fruits/seeds set, but F1 seeds inviable
PF = F1 hybrids partially fertile
HF = F1 hybrids highly fertile

The guide you are using isn't accurate as concerns anatomy of a flower. Stigma is the tip i.e. the hopefully receptive end of the style which is the long tube that the pollen gamete travels down through to arrive at the ovary. The ovary develops into the fruit we all love to eat called the chili. I could go on, but we have more important things to discuss.

 

[CaneDog]

I've wondered about the success rates in that study, as they seem low compared to anecdotal results. Despite using a dozen or so annuum varieties, they used only 3 baccatum, 2 chinense, and just 1 frutescens. The results on directional crossing and pre-crossing annuum x chinense into an AC x B cross are informative and I think have broad application when making these hybrids. I'd like to see the use of a greater mix of varieties within the B, C, and F species though, to see if the results hold up or perhaps support greater viability.

 

[PollenNut]

I've wondered about the success rates in that study, as they seem low compared to anecdotal results. Despite using a dozen or so annuum varieties, they used only 3 baccatum, 2 chinense, and just 1 frutescens. The results on directional crossing and pre-crossing annuum x chinense into an AC x B cross are informative and I think have broad application when making these hybrids. I'd like to see the use of a greater mix of varieties within the B, C, and F species though, to see if the results hold up or perhaps support greater viability.

I've wondered about the success rates as well as different studies using different plants within the same species to conduct the same experiment often get different results as well. Given how difficult it can be to cross pollinate some Capsicum in particular... I am just as inclined to look at such studies as guidelines rather than hard and fast rules and I make my crosses with the guidelines as examples by which to proceed. At the end of the day I will do my own thing and learn my own lessons firsthand rather than take any of the research as hard and fast facts. Greenleaf Tabasco is a very well known C. frutescens x C. chinense hybrid that always inspires me to make big changes sometimes it requires a lot of backcrossing and interline crossing. You can't expect to always hit a lottery winner in every single cross and definitely not right at the start.

 

[PollenNut]

I've wondered about the success rates in that study, as they seem low compared to anecdotal results. Despite using a dozen or so annuum varieties, they used only 3 baccatum, 2 chinense, and just 1 frutescens. The results on directional crossing and pre-crossing annuum x chinense into an AC x B cross are informative and I think have broad application when making these hybrids. I'd like to see the use of a greater mix of varieties within the B, C, and F species though, to see if the results hold up or perhaps support greater viability.

I also look at the fact that some people may be overestimating their ability when they guess at what went into their cross.... such guess work has led some to claim that Wartryx was a C. chinense x C. baccatum hybrid... which may indeed be true, but it also may not be true as well. I'd be more prone to believe that story if the bee would only be on speaking terms with us Homo sapiens when it made the cross.

 

[ahayastani]

I also look at the fact that some people may be overestimating their ability when they guess at what went into their cross.... such guess work has led some to claim that Wartryx was a C. chinense x C. baccatum hybrid... which may indeed be true, but it also may not be true as well. I'd be more prone to believe that story if the bee would only be on speaking terms with us Homo sapiens when it made the cross

A funny way of phrasing it, but definitely true.

 

[P.E.RSPEP]

Hello.
I wanted to share some stuff hoping to shed some light into the pungency thing.

Pungency is a quantitative trait, controlled by multiple genes. As such, it can be selected for or against.
The capsaicin synthesis pathway has many steps, and the factors affecting capsaicinoid accumulation have many regulatory genes we still haven't figured out.

But from what I understand.
The most widespread gene conferring absolute non-pungency it's " pun1 " as found only in annuum sweet pepper types.
This inhibits any expression of Capsaicin Synthase (CS) and thus no capsaicin is produced.

Nonetheless there are other mutations causing the complete dysfunction of this gene.

One of these dysfunctional CS alleles was named " pun1(2) " by the research team that found it in a C.chinense.

Genetic control of pungency in C. chinense via the Pun1 locus - PubMed

Capsaicin, the pungent principle in hot peppers, acts to deter mammals from consuming pungent pepper pods. Capsaicinoid biosynthesis is restricted to the genus Capsicum and results from the acylation of the aromatic compound, vanillylamine, with a branched-chain fatty acid. The presence of...

pubmed.ncbi.nlm.nih.gov

One of the researchers being Michael Mazourek, the developer of " Habanada ".
So most likely " Habanada " has this " pun1(2) " gene, and is the only widely available chinense with absolute zero heat.

There's also a " pun1(3) " found in C.frutescens. most likely the one responsible for the no-heat of " Numex NoBasco ".

But. There's also another well known mechanism by which peppers can lose pungency.
Some research proposed the name PUN2/pun2 but most papers talk about " pAMT ".

This gene affect a previous step in the capsaicin synthesis.
The conversion of vanillin to vanillylamine.

Some variants of this gene have been found to be partial or completely dysfunctional and cause an absolute loss of pungency.
There has been some research in annuum and chinense...
" Himo togarashi " it's the most available variety to have a non-functional " pAMT "

There's also some C.chacoense germplasm derives its lack of pungency due to this, a non-functional " pAMT " allele.

So this heatless chacoense could readily be used as pollen receptor to develop non pungent baccatum.
Although there's already some reportedly low-heat/heatless varieties " ají delight " and " CAP 455 ".
I ignore which mechanism is behind the loss of pungency of those two.

 

[armyofda12mnkeys]

Hello.
I wanted to share some stuff hoping to shed some light into the pungency thing.
....
One of the researchers being Michael Mazourek, the developer of " Habanada ".
So most likely " Habanada " has this " pun1(2) " gene, and is the only widely available chinense with absolute zero heat.

Thanks for that info, I actually got into contact with Michael. I'll send him my results at the end of the summer.
He said "The pun1^2 gene in Habanada is also in some other mild chinense. Some Aji Dulce for example."

 

[P.E.RSPEP]

Thanks for that info, I actually got into contact with Michael. I'll send him my results at the end of the summer.
He said "The pun1^2 gene in Habanada is also in some other mild chinense. Some Aji Dulce for example."

So cool you got in touch with Michael!

I've come across other paper describing another mechanism for loss of pungency. in Bolivian and Peruan peppers.

https://www.researchgate.net/publication/341628481_Multiple_Non-pungent_Capsicum_chinense_Accessions_with_a_Loss_of_Function_CaKR1_Allele_Originating_from_South_America

It'd be great to identify which gene each variety has as they become more available to the public.
From what I've read, at least the "strain 2 of ají dulce" has a mutated pAMT.

We could do test crosses with Habanada and Himo to find out.
What intrigues me the most are the supposedly heatless Brazilian varieties.

Also in the Numex Trick or treat, journal that can be found online. They seem to talk about another mechanism responsible for it's non-pungency.

It's crazy how things just get more and more complex the more you dive into. Aristotle was so right.

 

[armyofda12mnkeys]

Got my 1st ripe one... Looks a bit like last year's F1 parent... but yellow. I would say somewhere around 5 to 7 out of 10 on the heat scale (depending how hot your heat tolerance is hehe... for me its about a 5-6).
was a pleasant 3-4 on heat scale on the lower part and then 5-7 near the seeds...
Here is a pic of it, and compare to last years red F1 below. Very similar (but i would say has less heat).

Gotcha, thanks for the info!
Yeh the parents for this cross i think were pretty much very established named peppers (GrenadaSeasoning x Pérola Iaranja).
I've grown the former with my own seeds for years. I grew a few Pérola Iaranja last year... I will say one of the 4 P.I.'s was red vs usual salmon color. I'll have to look back at my notes to see which one was used for this cross.
All 6 successes I have all look like this (well 2 out of the 6 so far ripened red, and all 6 have the same shape):

Here are a couple more one of my growers got (he said about 7 out of 10 heat level):

Anyway, I got somewhere around 100-200 plants total from me and others that we growing ... so I'll post more pics soon .