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Gibberellic acid reversal, seeds Hermaphroditic.

gmanwho

Well-known member
Veteran
So 10 years ago i used Gibberellic acid on my Lavender plant. Collected the pollen. Hit 2 plants with the same pollen.

One plant hit was itself, a " lavender" plant we got from a san fran dispo like 2005ish. then i hit my bag seed plant Early pearl.

At the time the early pearl was a great plant, an 10 years later if i still had the original i can only speculate that it would hold its own today with the 20+ other lines im holding. at the least it is upsetting to myself an others that early pearl cut was lost. i have searched through a few leads with no luck


A few years ago when i grew out the lavender x lavender (R) plants they where all female, no actual lower balls. maybe a few top bananas.... top top cola bananas probably from high heat or light stress.

now i started the early pearl x lavender (R) 24 seeds , 22 popped. lost like 6 afterwards to damping off most likely caused from being over watered. A fungi like circle grew at the base of the stem an surrounding soil. The rest that made it grew healthy with no known problems. I just flipped 9 plants

so the first 4 plants showed ballls balls balls with no female pistols. I immediately thought how could this be to have full blown males.
I thought maybe i mislabeled the seed bag, or maybe i put some early pearl seeds i may have found during previous trims into the same bag. but i doubt i would have done that, in fact im pretty sure i would never do that an simply keep separate and label accordingly

the remanding 5 plants show female pistols and no balls. fast forward 20 days. 2 are now showing a few lower sacks.


is it possible the g acid levels carried over in the seed are enough to effect the offspring? if so could the clones possibly become more stable once the g-acid levels are lowered as the cut is grown out? Or is it just the early pearl genetics are not as stable compared to the first cross made?


thoughts? Have you experienced anything similar with g-acid, one cross was stable and the others not.
 
Well you had six die from damping off. You can't rule off stress from soil pathogens. That's almost 25% loss from disease, the rest definitely were affected
 

gmanwho

Well-known member
Veteran
Well you had six die from damping off. You can't rule off stress from soil pathogens. That's almost 25% loss from disease, the rest definitely were affected

Thanks for the reply. they did have a setback there. do you think the pathogens could have effected the plant months later even when they where growing happy and healthy?

i went back an edited the original post as i should have mentioned they could have been drowned out, which caused the damping off. at first when i originally posted , i was trying not to make the post over complicated and or to hard to follow with my poor writing skills.

what i should've said was that i had noticed my help had added to much water to the cups prior to them dying off. i had then placed the seedling cups on top of papertowels to help draw out water. it worked well an may have saved others but was probably to late. few days later some sort of fungus or bacteria started growing at the base of the stem. ghostly colored web growing in a circle pattern ontop the soil an growing up stem. like a circle tent.

i should also add all my mediums i run ewc teas. i also inoculate my soil or hydro mediums with mycos azo an trichoderma bacteria and fungi. a few others as well in the earthjuice biorighteous or recharge.

the plants where healthy the rest of their life, And today. im gonna let them ride some more as its an experimental room i built to go thru seedlings an hopefully find some keepers.

i went an inspected the 5 an removed all balls an lower larf sites. gonna keep an eye on them an see if new sacs appear.

i also plan on flowering one more clone of each asap, to see if those still carry. which i feel probably will still carry the intersex traits. just an experiment an something to learn at this point
 

slownickel

Active member
ICMag Donor
Veteran
Low nitrogen early on is supposed to cause pure males and hermies.

If your soil is to salted down to begin with, this stress alone will cause lots of males.

Are you reusing soil?
 
Low nitrogen early on is supposed to cause pure males and hermies.

If your soil is to salted down to begin with, this stress alone will cause lots of males.

Are you reusing soil?
So you're saying that plant sex is not determined at seed formation in the chromosome and that environmental factors contribute to sex selection?
 

gmanwho

Well-known member
Veteran
pretty straight forward case here. looks like u picked a seed out of a hermi plant ie bagseed.. often is the case these are hermi plants. u just got luck with ur cut but exploring the line u have found its hermi/intersex tendancys..

sounds like my most likely scenario. didnt even cross my mind that the early pearl was a herm to begin with.

appreciate every ones response's


still gonna flower out some smalls of 1 clone of each to see if it still carries. 2 didnt have any intersex, hoping keepers maybe?

i have like another 6 or 8 i haven't flowered yet. maybe a couple shots at another 2 or 3 stable keepers within those.

keep rocking!!!
 
Low nitrogen early on is supposed to cause pure males and hermies.

If your soil is to salted down to begin with, this stress alone will cause lots of males.
Although diecious, sexual expression in cannabis is not absolute. As we all know, hermaphrodites can sometimes be observed and stress can obviously play a part in their expression.

That being said, following fertilization the developing seed either be a female with two X chomosomes, or a male with an X and a Y. Nothing can happen from this point on that can change this fact. Even if a female is stressed to the point of appearing male, it is still a female.
 

clearheaded

Active member
good point NH, they are not true hermis but intersex. real hermis are like hemp grown around here with ruffly half of each type of flower.
 

slownickel

Active member
ICMag Donor
Veteran
Although diecious, sexual expression in cannabis is not absolute. As we all know, hermaphrodites can sometimes be observed and stress can obviously play a part in their expression.

That being said, following fertilization the developing seed either be a female with two X chomosomes, or a male with an X and a Y. Nothing can happen from this point on that can change this fact. Even if a female is stressed to the point of appearing male, it is still a female.

Yeah, happens to people too! haha..
 

Raho

Active member
Veteran
pretty straight forward case here. looks like u picked a seed out of a hermi plant ie bagseed.. often is the case these are hermi plants. u just got luck with ur cut but exploring the line u have found its hermi/intersex tendancys..

Not likely.
The Early Pearl produced no herms in offspring when crossed with my Mullum Super5 male selection.
That EP was a beast of a plant.
The Lav on the other hand was treated with Gib acid which is known to be a mutagenic. The gib treated Lav pollen is the most likely suspect for introducing herms in the offspring.

Remember though, each grain of pollen from that treated plant is unique, and assuming the Lav was not pre-disposed to sensitivity before the treatment, there is no reason to assume that just because some of the plants hermed on you from the Lav, that all of the Lav offspring will be vulnerable to that trait.
They might ;-) . . .
But more variability is to be expected than from an untreated parent.

All you need to do is find 1 killer stable plant brother G.
Not sure how many of those seeds you have, but the potential rewards of the cross are very high if you keep searching them.
 

gmanwho

Well-known member
Veteran
Not likely.
The Early Pearl produced no herms in offspring when crossed with my Mullum Super5 male selection.
That EP was a beast of a plant.
The Lav on the other hand was treated with Gib acid which is known to be a mutagenic. The gib treated Lav pollen is the most likely suspect for introducing herms in the offspring.

Remember though, each grain of pollen from that treated plant is unique, and assuming the Lav was not pre-disposed to sensitivity before the treatment, there is no reason to assume that just because some of the plants hermed on you from the Lav, that all of the Lav offspring will be vulnerable to that trait.
They might ;-) . . .
But more variability is to be expected than from an untreated parent.

All you need to do is find 1 killer stable plant brother G.
Not sure how many of those seeds you have, but the potential rewards of the cross are very high if you keep searching them.

i have a few hundred seeds, maybe 400... or so. 2 current clones express the ep look. thinner lime green leafs , more elongated bud structure compared to the original lav , trich coverage is there. but smells are sweet and flower like from the lav.

gonna run the clones an see. there potentially could have been a light leak in that room as well.

on another note i got 2 packs of the early pearl seeds from sensi in the mail.


next run the room will be tighter, sealed room with the co2 burner running, an def will be no light leak issues. so many variables...

thanks for stopping in raho
 

Darpa

Member
Hi Gmanwho,

Here is the explanation:
It could be that the Lavender plant that you treated with GA was not a true female (X/X) or that your early pear was not a true female (X/X)…

So in conclusion you ended up introducing a Xm/X allele in your project that is subject to hermaphrodism traits, therefore male inflorescence may be activated from environmental factors. It could be pretty easy to determine with strain introduced the Xm since you have lavender x lavender cross….

That subject was discussed in the (trick for controlling hormones on unstable strains) thread.

Here is some details:


The sexual phenotype of Cannabis often shows some flexibility leading to the differentiation of hermaphrodite flowers or bisexual inflorescences (monoecious phenotype). However this phenomenon is directly linked with both the plant genetic (chromosome), and the environmental condition. On an evolution point of view, the ability of a female plant to scent the absence of a male in its surrounding environment is not supported, since this evolution adaptation would have require the lack of male genetic in native plant population. Since sincemilla grow is new, on an evolution time scale, there would be no natural pressure for this new fitness phenomenon to appear as a new adaptation… Also, we should remember that Cannabis has a diploid genome (2n=20) with a karyotype composed of nine autosomes and a pair of sex chromosomes. A general belief suggest that Female plants are homogametic (XX) and male plants are heterogametic (XY). However, in these plants, sex expression is controlled by an X-to-autosome (X/A) balance system and not exclusively from the Y chromosome (confirmed from polyploidy essay)… The sex, is not determined by particular chromosomes, but by the genome assembly, and by the gene and chromosome interactions. All individuals are, in this manner, able to express one or the other sex. It was previously shown that sex expression also depends on different environmental factors and can be reversed by the application of plant growth regulators.

The following models were established:
- male plants with male inflorescence: X/Y, Xm/Y;
- male plants with female inflorescence, Xm/Xm;
- variable phenotypes, from true females to monoecious plants, but all with female inflorescence, Xm/X;
- female plants with inflorescence of female type, X/X

So if you get a clone only strain that may contain the Xm allele, you may end up with male inflorescence that may be activated from environmental factors…

So, in conclusion, if you don’t work with a true female X/X, you should pay good attention to the environmental stressor (light spectrum, plant density, temperature…etc) since all these variable will affect the predisposition of you plant to show intersex trait.

The Y chromosome will influence the sex determination of your plant, but the X\autosome ratio will also have an influence, specially with you female (hermaphrodite trait)...

Gender and Sexual Dimorphism in Flowering Plants, MONICA GEBER, Todd E. Dawson, Lynda Delph, Springer Science & Business Media, 6 déc. 2012 - 305 pages

https://www.icmag.com/ic/picture.php?albumid=71243&pictureid=1707831


Grant et al., 1994 concluded that the hemp has a sexual determinism based soon on X/autosomes equilibrium and not on a Y active mechanism.

Ainsworth, 2000 describes sex determination in the genus Cannabis as using "an X/autosome dosage-type.

Frankel and Galun, 1977 affirm that the males are X/Y, and the females are X/X. Besides that, the allele Xm exists, that determines the appearence of male flowers in female inflorescence. Thus, X/Xm plants have female inflorescence, but they can be not strictly females. Depending on genetic and non-genetic additional factors, these plants tend to masculinization, reason for which Kőhler named them “subgynoecious”. The Xm/Xm plants will have female inflorescence, but functionally are males. Considering the results obtained in various studies on polyploids by other researchers, Kőhler concluded that the masculinizing genes are autosomal and they are balanced by femaleness carrying genes of X chromosome, while the Y chromosome is “empty” under the aspect of sex determining genes.

An analogy could be made with Hop where polyploidy test confirmed the X\autosome relationship in sex determination:

Humulus lupulus possesses sex chromosomes. Female plants are homogametic with 2n¯18*XX, whereas male plants are heterogametic with either simple XY or multiple sex chromosome systems. Sex expression in hop appears to be regulated by an Xautosome balance system (Neve, 1961). This type of sex determination system is found in only two other genera, Cannabis and Rumex (Parker, 1990) and contrasts with the active Y-system found in some Silene species, in which the Y-chromosome specifies maleness (Westergaard, 1946, 1958). A ratio of the number of X-chromosomes to the number of sets of autosomes of 0.5 gives rise to a male plant; a ratio of 1.0 gives a female, while intermediate ratios give rise to intersexual individuals.

Sexual development and sex chromosomes in hop, H. L. SHEPHARD", J. S. PARKER#, P. DARBY$ C. C. AINSWORTH"*
"Plant Molecular Biology Laboratory, Department of Biology, Imperial College at Wye, Wye, Ashford, Kent TN25 5AH, UK #University Botanic Garden, Cory Lodge, Bateman Street, Cambridge CB21JF, UK $Department of Hop Research, Horticulture Research International, Wye, Ashford, Kent TN25 5AH, UK
Received 28 April 2000; accepted 7 July 2000

The Y chromosome is “empty” under the aspect of sex determining genes:

The X/autosomes equilibrium is responsible for the sexual determination of Cannabis and the Y chromosome is not directly linked in the process. The mechanism involve the presence of activators genes on the X chromosome and negative regulation encoded by the autosomes. In simple words, positive elements to activate a female developmental pathway exist on the X chromosome and these are balanced by element that favor male developmental program located on the autosomes. That mean that with the lack of an X chromosome in a XY plant the positive elements produced by the solitary X chromosome won’t be sufficient to counteract the effect of suppressor encoded by the autosome and you will end up with a male plant.

If the sexual determination of Cannabis would be based on the Y active mechanism concept, how would it be possible to induce a male diploid plant (XY) to produce female flower?

(Induction of female flowers on male plants of Cannabis Sativa L. by 2-chloroethanephos-phonic acid, HYM Ram, VS Jaiswal - Cellular and Molecular Life Sciences, 1970 – Springer)

Ok, since the majority of scientific research are performed on hemp vs Marijuana, I just want to state that the sex linked genes expression in female Cannabis flower (Drug cultivar) are similar to the one in the diocious hemp. That been said…,

There are several excellent recent reviews of sex determination of diocious plant and cannabis genus. I would suggest:

Boys and Girls Come Out to Play: The Molecular Biology of Dioecious Plants, Charles Ainsworth, Ann Bot (2000) 86 (2): 211-221.

The regulation of sex determining gene as you refer to is similar to the regulation observed in Silene latifolio. However, Cannabis show a different type of regulation:

The increased length of the Y chromosome in hemp (Yamada 1943 cited by Sakamoto et al. 1995) suggests some similarity with the sex chromosomes of Silene latifolia (Sakamoto et al. 1998; Fig. 2.5). In this latter species, males have XY and females XX sex chromosomes (Dellaporta and Calderon-Urrea 1993). The structure of the Y chromosome is characterized by a non-pairing region including sex-determining loci and a pairing, pseudoautosomal, region which undergoes recombination with the X chromosome during meiosis. The non-pairing region of the Y chromosome contains female suppressing, male promoting and male fertility regions (Westergaard 1953 cited by Di Stilio et al. 1998; Charlesworth 2002). The sex determinism in Silene is based on an Y-active system, with dominant male factors and female suppressing factors mapping to the Y chromosome (Dellaporta and Calderon-Urrea 1993). In contrast, in hemp, the sex determinism is based on a X-to-autosomes system. In addition, the genetic basis of sex determinism in Silene is strong and shows little evidence for lability or environmental effects (Ainsworth 2000), while hemp displays a high plasticity of sexual phenotype.


Here is a good PhD thesis on the subject:
QUANTITATIVE APPROACH OF THE GENETIC DETERMINISM OF SEX EXPRESSION IN MONOECIOUS HEMP (CANNABIS SATIVA L.), AND ITS RELATIONSHIP WITH FLOWERING PHENOLOGY AND STEM AND SEED YIELDS

https://dial.uclouvain.be/pr/boreal/...am/PDF_01/view

The sex determinism in dioecious hemp would be based on an X-to autosomes equilibrium and not on a Y-active mechanism (Westergaard 1958; Ainsworth 2000). This assumption agrees with the experiences carried out on polyploid hemp individuals (Warmke and Davidson 1944).

The X-to-autosomes system of sex determination is based on the ratio of the number of X chromosomes to the ploidy level. This system can explain why hemp polyploid individuals with XXY and XXXY formulae are female or female-hermaphrodite, and XY, XXYY and XYY individuals are male (Warmke and Davidson 1944).


The Y chromosomes are not necessary for the production of male flowers; moreover, the pollen can mature and be viable in the absence of Y chromosome.

The diversity of intersexual forms, the bipotency of sexually predetermined plants and the occurrence of fertile male flowers on female plants of dioecious hemp suggested that the sex in hemp would be determined by the activity of genes that are located not only on the sex chromosomes but also on the autosomes (Grisko 1937 cited by Truta et al., 2007; Sengbusch, 1952 cited by Westergaard, 1958; Rath, 1968 cited by Truta et al., 2007; Migail, 1986 cited by Mandolino and Ranalli, 2002). Grişko (1937 cited by Truta et al. 2007) considers that the determinants of phenotype and those of sexualisation are independent since female plants are able to produce male flowers under given environmental conditions. The ability to reverse the sex determination mechanism by hormonal treatment suggested that sex determination genes could regulate alternative programs of sexuality through a signal transduction mechanism that modifies endogenous hormonal levels (Dellaporta and Calderon-Urrea 1993).



Markers, referred to as MADC, for male-associated DNA sequence in C. sativa:

Hybridization of DNA from male and female plants by southern blotting analysis revealed either the absence of no sex-linked polymorphism (Mandolino et al. 1999), either multiple bands among which only few are male-specific (Sakamoto et al. 2000; Torjek et al. 2002; Sakamoto et al. 2005). Similarly, the in situ hybridization of chromosomes with the MADC3 and MADC4 sequences produced signals dispersed on all chromosomes and thus not specific of the Y chromosome. Thus multiple sequences encoding retrotransposable elements should exist ubiquitously in the genome of C. sativa (Sakamoto et al. 2005). This appears in agreement with the view of Clark et al. (1993), who observed no major difference in the distribution of repeated DNA sequences between X, Y and autosomes in Rumex acetosa, a dioecious species with heteromorphic sex chromosomes. According to Charlesworth (2002), the abundances in repeated DNA in plants bearing sex chromosomes would be mostly similar on both sex chromosomes and autosomes.
 

GMT

The Tri Guy
Veteran
Hey, OK in response to your pm asking for my input on this, here it is:

G.A. is, as others have pointed out, a mutagen. This means that any growth that occurs after being exposed, may (but not guaranteed) possess alterations to the genome that the plant started with. It is not normally used as a reversing agent by those wishing to reverse a female, for the purposes of creating feminised seeds. The preferred choices being STS AND CS. It is however a good choice for the creation of something new.
The problem with evolution however, is that any changes are normally bad for the inheritor of those changes. It is rare to find a beneficial change. The normal method is to soak premade seeds in a GA solution, and then sprout the seeds. When treating a plant, the stress caused to the plant can have many effects upon the plant. This stress, will, over time, subside; but the alterations to the genome will not.
Offspring from this event, have any number of possibilities. An alteration to the genome is not guaranteed, and as said, any present is unlikely to be beneficial. But this does not rule out the possibility. One common noted benefit is the the creation of tetraploid genomes, which, in the event of exclusively female parentage, should not cause the confusion over sexual determination that seems to have been confusing botanists for decades. However, that being said, any mutation in any of the systems involved in the expression of flower formation, could easily exhibit themselves in intersexed flowers. Whether or not that is the case will be plant specific, not event specific, meaning just because one plants looks like a Hermie, doesn't relate to the odds of the next plant being the same. Also it should be noted, that such an alteration, may result in one flower type being non functional ie. Balls may not produce functional pollen.
Numbers are key to this type of breeding since this is a case of searching through the mutants to find the superheros.
Hope I covered what you asked me for, but I feel I'm waffling now, so ask and I'll try to answer if I missed something.
 

Darpa

Member
Hi GMT!
G.A doesn't have any mutagenic effet on plant. It's a plant hormone and not a mutagen. Therefore it doesn't induce DNA alteration…I could provide you with scientific research publication on the subject if you want.

The fact that the plant were reversed with G.A is irrelevant in this case.
 

Raho

Active member
Veteran
So Weird. I know I did a quick search and confirmed my post before I hit submit, but now can't find any links to support GA3 as a mutagenic agent in plants.

I wonder Gman, were either of the 2 parent lines ever exposed to colchicine?
 

Darpa

Member
Colchicine is a known mutagen and act as a antimitotic agent. It's the principal chemical I used to induce polyploidy in cannabis (seeds treatment but mainly tissu culture). However, colchicine doesn't induce sex reversal like G.A or Silver Thiosulfate.

I don't see why colchicine would have been use in this project. Gman will probably confirm that info….

Cheer!
 
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