Male clones transformed to Female to judge male smoking quality

[Tom Hill]

I believe we have to classify selfing as sexual reproduction. Yes asexual reproduction is a shortcut, a very attractive alternative. Identical reproduction will never be scoffed at by the serious. It seems its worth is greatest not with IL's or pure lines, but with plants that are heterozygous for many loci, exploiting heterosis to a maximum.

Are you saying that if you were charged with producing acres/hectares of high quality drug cannabis that you would not be using clones? Truthfully?

If we can clone, or double haploids on the instant, then where is the advantage of sourcing this material from highly bred populations? -Tom

 

[Unclepeter]

Well I'd love to be able to have DH,s at the ready myself...most of the breeding material I have read is to do with corn though... I'd honestly love the availability of artificial seeds but will we ever see them?

http://www.springerlink.com/content/7560073j08746340/
http://resources.metapress.com/pdf-preview.axd?code=w317822x7t27u413&size=largest
http://books.google.com/books?hl=fr...roduction&ots=Q-1J9tptq5&sig=VHtwfNzXcO75LsBN

but what we see at the moment IMO with self seed is the release of inbreds that are then selfed... but we are going to get to the point of no more males again and that in itself leads to division etc...

And I clone of course!.

It seems its worth is greatest not with IL's or pure lines, but with plants that are heterozygous for many loci, exploiting heterosis to a maximum.

now this bit has my interest.... I find more like that!!!

 

[Unclepeter]

haha you know I like a good jab, I'd love a discussion on DH myself, I know theres a chap on here called spurr who has an interest in them....

my use for them would to be as parents (least that was my thinking!)...

 

[Tom Hill]

Artificial seed would be great, a huge factor that if solidly in place would reshuffle the deck considerably imo. Though not there yet, we best leave room for that in the future.

Regarding what we're seeing at the moment in the breeding of cannabis, good lord that's too easy to pick apart.

What I'm attempting to do is put into check those arguing solely from the corn breeders set of realities. To give a moment of pause to some, and they know who they are - I'm as guilty as anybody there. To point out that several key factors are being overlooked when we try to carbon copy cannabis breeding from that of corn. That when reviewed and revised to truly suit cannabis, we are very likely to be arguing method from some other chair than solely that of the maize breeder - or should be anyway..

 

[GreenintheThumb]

to me DH is the future

Why?

 

[spurr]

haha you know I like a good jab, I'd love a discussion on DH myself, I know theres a chap on here called spurr who has an interest in them....

my use for them would to be as parents (least that was my thinking!)...

FWIW, I have worked out my basic protocol for creating DH cannabis, male and female. I will be running trials after I fine-tune the protocol and TC protocol, etc. I should be able to create the first DH drug cannabis by years' end, if everything works. I too am using it for parents.

 

[spurr]

to me DH is the future

Why?

To make nearly 100% percent homogeneous F1 hybrids, no need to make F5, F6, etc., the F1 will be more homogeneous than an IL; and to use it make DH from clone-only 'elites', etc.

One can also use DH breeding with selfing methodologies, but that is different than using DH for parents to make nearly 100% homogeneous F1.

I plan to start breeding projects offering the only DH sourced and homogeneous F1 seeds on the marketed...offering both nearly 100% homogeneity and hybrid vigor (that is not found in F2, F3, F4, etc.)...no more poly-unstablized hybrids!

 

[GreenintheThumb]

Okay.

Still not really following you. How do you plan to induce chromosome doubling? And then when you do how do you know which seed is successful? Again, I'm a corn minded dumb shit so that's mostly what I know (hi, Tom ) but maybe you're going to do it in vitro?

Have you ever done this successfully?

And I don't really understand how they'd be considered an F1 if what I think you're offering is a clone only in seed form...I think I'll just shut up until you expound on your methodology.

 

[Unclepeter]

Artificial seed would be great, a huge factor that if solidly in place would reshuffle the deck considerably imo. Though not there yet, we best leave room for that in the future.

Regarding what we're seeing at the moment in the breeding of cannabis, good lord that's too easy to pick apart.

What I'm attempting to do is put into check those arguing solely from the corn breeders set of realities. To give a moment of pause to some, and they know who they are - I'm as guilty as anybody there. To point out that several key factors are being overlooked when we try to carbon copy cannabis breeding from that of corn. That when reviewed and revised to truly suit cannabis, we are very likely to be arguing method from some other chair than solely that of the maize breeder - or should be anyway..

guilty as charged with corn....

 

[Unclepeter]

Why?

speed and relation to genetic advance..... I'd do at least 7 rounds of inbreeding with this its 2. but again I have been looking at the corn boys ie pioneer etc... you could be knocking out hybrids yearly( I think)

my first interest came from reading a fella called A Gallais

 

[Unclepeter]

FWIW, I have worked out my basic protocol for creating DH cannabis, male and female. I will be running trials after I fine-tune the protocol and TC protocol, etc. I should be able to create the first DH drug cannabis by years' end, if everything works. I too am using it for parents.

glad you are here.. (also should this now split from this thread?)

 

[Unclepeter]

Okay.

How do you plan to induce chromosome doubling? And then when you do how do you know which seed is successful? Again, I'm a corn minded dumb shit so that's mostly what I know (hi, Tom ) but maybe you're going to do it in vitro?

from what I gather some use colchicine others like you say go for vitro...

 

[spurr]

Okay.

Still not really following you. How do you plan to induce chromosome doubling? And then when you do how do you know which seed is successful? Again, I'm a corn minded dumb shit so that's mostly what I know (hi, Tom ) but maybe you're going to do it in vitro?

Have you ever done this successfully?

No I have not yet attempted it, but the protocol is sound, albeit the protocol will need fine-tuning and testing. I will be working on this project with a buddy this year, starting in a few months, he has a degree in plant breeding/genetics, so we should make a good team to work out any kinks.

And I don't really understand how they'd be considered an F1 if what I think you're offering is a clone only in seed form...

It's an F1 because you breed two different DH parents together (they can be two different cultivars or races; or the same cutlivar or race), just like breeding two different diploid parents together gives you an F1.

I do not plan on using DH inducers, like is done with corn, etc.

I think I'll just shut up until you expound on your methodology.

Sorry, but I won't be sharing my protocol anytime soon, it will be proprietary for the foreseeable future. But I do plan on offering seed to market...that way a grower won't need n seeds to find a 'keeper'; every seed should be very similar to each other, thus all should be keepers

All this is contingent upon me being able to product DH cannabis, it's not a sure thing.

 

[spurr]

FWIW, I have worked out my basic protocol for creating DH cannabis, male and female. I will be running trials after I fine-tune the protocol and TC protocol, etc. I should be able to create the first DH drug cannabis by years' end, if everything works. I too am using it for parents.

glad you are here.. (also should this now split from this thread?)

Thanks, I hope we can let bygones be bygones. I am not sure if this should be split, I haven't read this thread yet, I only noticed you mentioned me and DH, so I thought I should post an update on my work.

I do not plan on providing info on how to make DH cannabis to the public at this time. But, to the degree I can assist in a DH specific thread, I will assist.

 

[GreenintheThumb]

I do not plan on using DH inducers, like is done with corn, etc.

Well that answers part of my questions. If you're willing to speak more on your planned methodology please pm me. I'd like to waste a lot of your time.

All this is contingent upon me being able to product DH cannabis, it's not a sure thing.

Best of luck to you. The future is bright for our beloved plant.

 

[GreenintheThumb]

I think we should split this into a new thread.

Doubled Haploid Cannabis perhaps? Where the mods at? Or Sam, it's his thread.

speed and relation to genetic advance..... I'd do at least 7 rounds of inbreeding with this its 2. but again I have been looking at the corn boys ie pioneer etc... you could be knocking out hybrids yearly( I think)

Will you expound upon where you got the number 7?


And from what I've read (again in corn, damn have I got a one track mind) DH techniques weren't very more effective than traditional selfing. Any thoughts on what makes this so useful for cannabis?

Tom, I saw a while back you said DH was more useful in crops that aren't propagated by cutting. Can you expand on how you came to that conclusion?

 

[spurr]

I do not plan on using DH inducers, like is done with corn, etc.

Well that answers part of my questions. If you're willing to speak more on your planned methodology please pm me. I'd like to waste a lot of your time.

Glad to help clear up at least a little up for you, sorry I am being evasive. I have put a lot of time and effort into my quest, so I don't want to give it all away, yet. I do plan to make the protocol public at some point, maybe in a peer-reviewed paper, if the protocol(s) work(s) and the paper is accepted...

Best of luck to you. The future is bright for our beloved plant.

Thanks, I think lots and lots of people will benefit if I can work it out. I am also working on QH (quadrupled-haploid) cannabis, which will be simple if I can make DH. QH cannabis is intriguing to me, just like 4n (tetraploid) cannabis is intriguing

 

[GreenintheThumb]

Another thing I forgot to ask.

FWIW, I have worked out my basic protocol for creating DH cannabis, male and female.

Out of curiosity what makes you want to work diecious lines as opposed to gynoecious ones? Was this a conscious choice or more a reality due to your protocol?

I know you were called out and showed but what're your thought on the OP subject? Are reversing males worth the time and effort? I think it's novel and interesting and could lead to further understanding of the expression of cannabis flowers in a scientific context. But for the most part reversing your males and selecting the best one or two by smoke seems kind of dumb to me.

Sam-
Do you have any updates to this thread? Any pictures of your reversals or attempted reversals?

 

[spurr]

Another thing I forgot to ask.

Out of curiosity what makes you want to work diecious lines as opposed to gynoecious ones? Was this a conscious choice or more a reality due to your protocol?

I plan to work with both types of lines. Using female-only breeding (e.g., selfing), if that is what you are asking, would mean less work because I would only need to prefect one protocol to create DH plants. I want to use both F/M breeding and F/F breeding (incl. selfing and non-selfing) to see what path gives better results. I kind of like the idea of F/M over F/F because it's the natural state for cannabis, but I plant to take both roads and see where they lead...

I know you were called out and showed but what're your thought on the OP subject? Are reversing males worth the time and effort?

Sure, I think so, no need to rule anything out IMO. I haven't looked into it, but I plan to after I read this thread. Making male DH plants should be easier (higher chance of success) than female.

I think it's novel and interesting and could lead to further understanding of the expression of cannabis flowers in a scientific context. But for the most part reversing your males and selecting the best one or two by smoke seems kind of dumb to me.

Ditto

 

[highonmt]

What you have done is to breed with an inter-sexed male. This will pass on inter-sexed genes to the progeny.
-SamS

Sam,

First thanks for posting all this fantastic information, you may want to wear old pants if you are smoking too much of this reversed male herb her's a quote from the msds these effects happened upon oral dosage
" Subjects receiving the test compound reported the following symptoms and/or signs; sudden onset of diarrhea or an urgency of bowel movements, stomach cramps or gas and increased urgency or frequency of urination."

However the degradation of chloro phosponates like this are really quite fascile and the etylene is of course a gas so it won't last long at all on the plant.

OK so I read most of this thread and I really think that what people are seeing regarding "intersex genes" and chemical intersex conversion is not the result of classic XY chromosomes which are very rare in plants in general an even more so in dioecious plants and do not exist afaik in subdioecious cannabis. AFAIK the male chromosome in cannabis has not been reported to exist. I've read a bunch of recent papers on the subject and every one of them imply that cannabis sexual expression is of the X/autosomal dosage type or X/ male genetic marker type. That is what makes cannabis so sexually labile that experiments of the type indicated in this thread are possible. The YY cannabis of which you speak simply is not real IMO and in the opinion of all the scientific writers I've found so far. If you have seen a paper indicating otherwise please post it. Subdiecious plants are not well understood but there are only a handful that have xy chromosomes and those are concidered recent evolution. The reason I post this is because I have not seen any posts indicating that etylene is an inhibitor of androgenic hormone production and is not changing anything else. The female potential in cannabis is universal the maleness in cannabis comes about because of the dominant expression of the autosome/ genetic markers that control sexuality. If this plant had an XY chromosomal arrangement the reversal seen in the presence of ethylene would not occur to the degree we see, A very logical and common example of the X/autosomal situation in cannabis is environmental stress; If a female cannabis plant is stressed the production of androgen hormones is initiated in a response theorized to protect the viability of the species and walla monoecious individuals (intersex plants) result. Stress a male and you still have a male. Some males do seem to have subdominant male genetic expression and will allow seepage of female expression to occur. Selecting for this subdominant trait seems to lessen the occurance of monecious individuals in it's progeny... anyway just been doing research for upcoming projects so thought I'd chime in. Thanks to all who posted on this thread and to Sam for all the great info and experience you share here.
HM
Well this is what I found on springerlinks not 5min after posting the above...turns out you might be right Sam and a lot of other researchers may be wrong...or maybe they all are wrong or maybe they are just all right. Who knows I'm enjoying reading the papers tho lots of interesting insight into cannabis genetics here:http://www.springerlink.com/content/0014-2336/?k=cannabis

Here is a link to the below...http://www.springerlink.com/content/t491042240h422v8/

Cannabis sativa L. is a dioecious species with sexual dihttp://www.springerlink.com/content/t491042240h422v8/morphism occurring in a late stage of plant development. Sex is determined by heteromorphic chromosomes (X and Y): male is the heterogametic sex (XY) and female is the homogametic one (XX). The sexual phenotype of Cannabis often shows some flexibility leading to the differentiation of hermaphrodite flowers or bisexual inflorescences (monoecious phenotype). Sex is considered an important trait for hemp genetic improvement; therefore, the study of the mechanism of sexual differentiation is of paramount interest in hemp research. A morphological and molecular study of Cannabis sativa sexual differentiation has been carried out in the Italian dioecious cultivar Fibranova.Microscopic analysis of male and female apices revealed that their reproductive commitment may occur as soon as the leaves of the fourth node emerge; the genetic expression of male and female apices at this stage has been compared by cDNA-AFLP. A rapid method for the early sex discrimination has been developed, based on the PCR amplification of a male-specific SCAR marker directly from a tissue fragment.

Five of the several cDNA-AFLP polymorphic fragments identified have been confirmed to be differentially expressed in male and female apices at the fourth node. Cloning and sequencing revealed that they belong to nine different mRNAs that were all induced in the female apices at this stage. Four out of them showed a high degree of similarity with known sequences: a putative permease, a SMT3-like protein, a putative kinesin and a RAC-GTP binding protein.


and this from 2005
DNA from female and male hemp (Cannabis sativa L.) plants belonging to nine different varieties were screened with180 RAPD primers in a search for sex-associated DNA markers. About 1500bands were produced in total, nine primers were found yielding one or two DNA bands amplified in all nine male DNA bulks and absent in all female DNA bulks. These putatively male-associated markers were then scored in three different F1progenies, deriving from a cross between a common male parent and three different female plants. The sex of the progeny was accurately scored on the basis of the floral phenotype, and the presence of the nine male-associated markers was verified by RAPD analysis. In all three progenies examined, all the male plants showed the DNA markers previously identified by bulk segregant analysis (BSA) on the hemp varieties, while all the female plants lacked them. The fact that the association between these markers and the staminate phenotype is found when examining male plants of distantly related varieties, and that such linkage is never broken when different progenies are examined, strongly supports the hypothesis that the markers found are physically located on the Y chromosome, in a region excluded from recombination during meiosis. Another marker was shown to be present in the male parent, in all the male plants of each progeny, and in 50% of the female progenies, while it was absent in the female parent; the possible occurrence of markers deriving from multiple amplification sites of the genome is discussed.