I have a topic of discussion maybe y'all can help me with

  • Thread starter MagicNative1
  • Start date
  • Tagged users None
MagicNative1

MagicNative1

109
28
I recently started a new grow a few months ago with some basic bag weed seed and went through probably 30 seeds just to get my main 2 plants and then a back up for both ..
Popped a seed and a back up... 2 weeks later popped another and a back up...

Eventually a week before flower ( I did the entire 9 weeks of veg ) I started to see pre sexing starting to happen which let me know I was pretty much right on time. But the first to start showing was a male about 2 weeks ago, lil less probably...
And last week after I switched to 12 and 12... ( Probably going to drop it down to 11 so I can run my other tent, and to much power draw all at the same time throws my breakers repeatedly...

Anyways, so I ended up finding 2 more males... Now I've just got my female and a smaller clone tent with 7 clones of the males and have just switched to my second step cloning phase before I end up just taking the dip off to dry them out and for better lighting.. ( I still mist them several times a day after this point too ) but anyway I already cleaned up about 40% or so of her leaves last week and tightened up the rest of her tonight cleaning out between nodes and everything up until just above where my first half of lollipoping raised her foliage up to last week... Then I went to tying her back down to spread her back out.. I untied everything a week or 2 before switchin over to let the plant have some release and let her try to decide her own path for a little while so the tie down would be another form of last when I started her up again..

Any so now finally to the part I need some more insight on... And I'm pretty sure not just everyone is going to know a legit answer to this question... In fact I'm pretty sure I already know most of the answers I'll get which is probably going to be.... Don't do it... But this is something legit I've heard a few times from some big names in our grow society... But upon doing the research to try and find out truths behind it online or even over YouTube I have literally not found even one more insight onto the topic..

So here it goes...
I have heard that when a female is in flower and knows there's no way she's going to be pollinated out in the wilds that she will often Hermie just to self pollinate and provide a future offspring..
This I've heard alot and most growers know... Others know of Hermie's and just know that's not what they want... 🤷🏻‍♂️

But what I've been hearing a few times in this last 3 or 4 months right before I started the grow and onwards... Like I said maybe 3 or 4 times at least I've heard this and it is:
That a female that knows it's near a male will also act similar to how a male does (by genetics and by choice) by trying to grow Fuller get bigger and faster because it also wishes to be pollinated... Males do it instinctually because like a male human they want to be bigger n have their chest poke out all big n strong n be the first to get the girl...

But is it true:
That in the presence of a male plant females might also similarly do the same thing......


🤔🤔🤔

Cuz I have a tent with at let another 4 males and as long as there not progressing to far I thought why not just take the furthest one along and instead of just shredding it why not put it back in with the female for another 2 maybe 3 days then shred it.. give it a day or 2 by herself and then replace with another male n so forth until they are gone...

I already know seeds are bad, lol definitely don't need more seed in a bag seed collection off the street and no genetics behind it...

But will it work or not... Should I test the theory and chance it or would you as well if this information was something you thought could be possible trash them and not give it the shot...

Let me know what you think

And what you think you'd do if presented with this knowledge and opportunity to try,

I'm not going to screw her up she doesn't have to much of a leg on it so if I don't get enough indefinite answers sayin hell yeah I've heard that or hell yeah I'd try it for sure or something I'm probably going to trash them anyways, but I literally have days to decide these males are pushing even without switching their light cycles.. lol
 
Moshmen

Moshmen

Staff
Supporter
6,633
313
I recently started a new grow a few months ago with some basic bag weed seed and went through probably 30 seeds just to get my main 2 plants and then a back up for both ..
Popped a seed and a back up... 2 weeks later popped another and a back up...

Eventually a week before flower ( I did the entire 9 weeks of veg ) I started to see pre sexing starting to happen which let me know I was pretty much right on time. But the first to start showing was a male about 2 weeks ago, lil less probably...
And last week after I switched to 12 and 12... ( Probably going to drop it down to 11 so I can run my other tent, and to much power draw all at the same time throws my breakers repeatedly...

Anyways, so I ended up finding 2 more males... Now I've just got my female and a smaller clone tent with 7 clones of the males and have just switched to my second step cloning phase before I end up just taking the dip off to dry them out and for better lighting.. ( I still mist them several times a day after this point too ) but anyway I already cleaned up about 40% or so of her leaves last week and tightened up the rest of her tonight cleaning out between nodes and everything up until just above where my first half of lollipoping raised her foliage up to last week... Then I went to tying her back down to spread her back out.. I untied everything a week or 2 before switchin over to let the plant have some release and let her try to decide her own path for a little while so the tie down would be another form of last when I started her up again..

Any so now finally to the part I need some more insight on... And I'm pretty sure not just everyone is going to know a legit answer to this question... In fact I'm pretty sure I already know most of the answers I'll get which is probably going to be.... Don't do it... But this is something legit I've heard a few times from some big names in our grow society... But upon doing the research to try and find out truths behind it online or even over YouTube I have literally not found even one more insight onto the topic..

So here it goes...
I have heard that when a female is in flower and knows there's no way she's going to be pollinated out in the wilds that she will often Hermie just to self pollinate and provide a future offspring..
This I've heard alot and most growers know... Others know of Hermie's and just know that's not what they want... 🤷🏻‍♂️

But what I've been hearing a few times in this last 3 or 4 months right before I started the grow and onwards... Like I said maybe 3 or 4 times at least I've heard this and it is:
That a female that knows it's near a male will also act similar to how a male does (by genetics and by choice) by trying to grow Fuller get bigger and faster because it also wishes to be pollinated... Males do it instinctually because like a male human they want to be bigger n have their chest poke out all big n strong n be the first to get the girl...

But is it true:
That in the presence of a male plant females might also similarly do the same thing......


🤔🤔🤔

Cuz I have a tent with at let another 4 males and as long as there not progressing to far I thought why not just take the furthest one along and instead of just shredding it why not put it back in with the female for another 2 maybe 3 days then shred it.. give it a day or 2 by herself and then replace with another male n so forth until they are gone...

I already know seeds are bad, lol definitely don't need more seed in a bag seed collection off the street and no genetics behind it...

But will it work or not... Should I test the theory and chance it or would you as well if this information was something you thought could be possible trash them and not give it the shot...

Let me know what you think

And what you think you'd do if presented with this knowledge and opportunity to try,

I'm not going to screw her up she doesn't have to much of a leg on it so if I don't get enough indefinite answers sayin hell yeah I've heard that or hell yeah I'd try it for sure or something I'm probably going to trash them anyways, but I literally have days to decide these males are pushing even without switching their light cycles.. lol
Bro science ! Ditch the males unless ya want a bunch of seeds
 
MagicNative1

MagicNative1

109
28
Bro science ! Ditch the males unless ya want a bunch of seeds
Was definitely believing that myself... But I've heard it from people on YouTube / podcasts I listen... Like say Mr.Growit and one or 2 others from someone they had on the show.. And when I heard it more than once I was like well this isn't something you seen your buddy posting about on fb or heard from the greatvine, it's legitimate people within the community N stuff... And I'm not really saying they spent like 30 min - an HR discussing it but they'd mentioned it...

And I'll admit in a way if you get into depths of stuff like social interaction between plants or even during flowering the way they respond to things it kinda in a way makes sense... But not like enough where I'm like yup.... Facts my man... Lmao.. cuz really I don't know... I don't have a clue.... But if a single female can switch things up and seed herself knowing nothing is around to further the production to me sounds like a chemical response to her surroundings.. knowing what she's gonna have to do to press on... So why not something similar just off a different basis...

But yeah I feel you in the back of my head I can definitely hear that voice screamin BRO SCIENCE... 🤣

And like I said I don't want to have any chances of pollination..

But.......

I also went and put this guy in with her last night just to free up some space in my clone tent had a little to much in there... Lol

He doesn't really even have anything goin for him yet, his first crab claws are just coming out and honestly probably tomorrow night depending on how he's progressing I am probably trashing him just in case and switching him out for another pre flower male just around the corner from full on pollen production. I'm seriously not even watering the males anymore.. haven't for a few days... After this next switch the 3rd ready male will probably already be dying before I get him in.. 🤷🏻‍♂️ which is cool with me too... But... Just thought trying it might be cool..
 
IMG 20220124 005445
IMG 20220124 005455
MyGrowYo

MyGrowYo

261
63
Was definitely believing that myself... But I've heard it from people on YouTube / podcasts I listen... Like say Mr.Growit and one or 2 others from someone they had on the show..
This is how bro science gets started imo. Sure mr.growit isn’t an idiot, and has some really interesting guests on his podcast, but he’s not a horticulturist or biologist. He’s just another grower, like you or me. Ill wait for someone like Dr. Bruce Bugbee to present some real science on stuff like this, after a peer review.
 
MagicNative1

MagicNative1

109
28
This is how bro science gets started imo. Sure mr.growit isn’t an idiot, and has some really interesting guests on his podcast, but he’s not a horticulturist or biologist. He’s just another grower, like you or me. Ill wait for someone like Dr. Bruce Bugbee to present some real science on stuff like this, after a peer review.
Haven't heard of them... Cool something new to learn about... Welp it didn't matter anyway she/he developed alot more and she was a boy anyways 🤣... Tried popping over 30 seeds and had gotten those 4 plants only to get all boys..
 
IMG 20220124 191821
sambapati

sambapati

1,203
263
Haven't heard of them... Cool something new to learn about... Welp it didn't matter anyway she/he developed alot more and she was a boy anyways 🤣... Tried popping over 30 seeds and had gotten those 4 plants only to get all boys..
If you want to increase the likelihood of having female plants there are specific protocols to follow: including at least a 4" high container, temperature control, humidity control and so forth. In my current grow I got 9/10 females from seeds by falling all the protocols exactly as they are specified. -- increase nitrogen, higher humidity, fewer light time and more dark, use lots of blue light and less stress (ambient light temp variations). The more you know -- the beta you grow.
 
ComfortablyNumb

ComfortablyNumb

5,770
313
If you want to increase the likelihood of having female plants there are specific protocols to follow: including at least a 4" high container, temperature control, humidity control and so forth. In my current grow I got 9/10 females from seeds by falling all the protocols exactly as they are specified. -- increase nitrogen, higher humidity, fewer light time and more dark, use lots of blue light and less stress (ambient light temp variations). The more you know -- the beta you grow.
I was just reading a study that said the seed has it's gender when it's initially grown. Environment has no real effect on gender. You will hear otherwise, but its wrong. What environment can do is over stress a plant and cause it to mutate and hermie.

 
Last edited:
ComfortablyNumb

ComfortablyNumb

5,770
313
Lets imagine an outdoor grow for example. Same environment, where do the males come from if environment can affect gender? If that was the case, they would all be male.
 
sambapati

sambapati

1,203
263
I was just reading a study that said the seed has it's gender when it's initially grown. Environment has no real effect on gender. You will hear otherwise, but its wrong. What environment can do is over stress a plant and cause it to mutate and hermie.

CN -- just reading some of my jorge cervantes medical growers bible:

Environmental factors start influencing sex the moment the seedling has three pairs of true leaves not including the cotyledons ( first set of 2 true leaves). Enviromental factors that influence sex determination of cannabis include but are not limited to:
- Increasing the level of nitrogen makes more female plants, lower the nitrogen level to creat more male plants, increase the level of potassium will increase male tendencies. lower the potassium level will encourage female plants, in other words a higher nitrogen level and a lower potassium level for the first two weeks increses female.
- Lower temperatures increses the number of females, and higher temps (warm temps) will increse male growth.
- High humity increses the number of female plants, and low humidty increases male plants as well.
- Low growing medium moisture increases males.
- More blue light will increses the number of female plants, and more red light increses male tendenciese.
- fewer hours of light will increse the number of females, and loonger days for plants with more hours of light will increse male plants.
- Stress - any enviromental stress tends to yeild more male plants when growing from seed,
- Ofcourse when buying female seeds, these factors count eachother out, unless the enviroment is like stated above, that will increse make tendencies.

So we're talking specifically about what could potentially alter the results of the seeds being planted in the ground/media to try and increase the number of females.

This topic is many years old on this forum and others. All I know is 9/10 females and 7/7 from my last grow after following all the above to the letter. Remember K-Mart's Blue Light Special? If we (farmer) are aiming our blue lights at the seedling -- we're creating an effect not found in 'nature' -- . I'm not an expert....don't even play one on TV. And remember IBM's Chairman saying "There will never be a need for more than 4 computers" or Digital's Ken Olsen saying, "You would never need a computer in the kitchen." Oops...both leaders, both inventors and both wrong.
 
ComfortablyNumb

ComfortablyNumb

5,770
313
How do you explain males in the wild?
Logically, the fact that environment has a major effect on plant gender is a fallacy. It can not happen, at least not the way we have been discussing it so far.

Ok, its in his book. What evidence does he give?
 
sambapati

sambapati

1,203
263
N
How do you explain males in the wild?
Logically, the fact that environment has a major effect on plant gender is a fallacy. It can not happen, at least not the way we have been discussing it so far.

Ok, its in his book. What evidence does he give?
Not trying to explain males in the wild or nature -- just trying to help this one grower using my own personal experience that says these things might work. The same information is in Bergman's book. If you don't believe it it doesn't matter what 'evidence' is presented. We'll never get to the moon, right? There will never be animal or human clones right? All these notions proved false.

The purpose of this site is to help others with their problems and share what works, or in my own limited view to make the world an easier kinder place. Thats all.

I Love Growing MJ by Robert Bergman

Cannabiz Sativva


Cannabiz Sativva·Check out the site I Love Growing Marijuana for all your Marijuana and Cannabis needs. Growing Cannabis, Marijuana Grow, Growing Indoors, Indoor Growing, Grow Pot, Grow Marijuana.
 
ComfortablyNumb

ComfortablyNumb

5,770
313
N

Not trying to explain males in the wild or nature -- just trying to help this one grower using my own personal experience that says these things might work. The same information is in Bergman's book. If you don't believe it it doesn't matter what 'evidence' is presented. We'll never get to the moon, right? There will never be animal or human clones right? All these notions proved false.

The purpose of this site is to help others with their problems and share what works, or in my own limited view to make the world an easier kinder place. Thats all.

I Love Growing MJ by Robert Bergman​

Cannabiz Sativva


Cannabiz Sativva·Check out the site I Love Growing Marijuana for all your Marijuana and Cannabis needs. Growing Cannabis, Marijuana Grow, Growing Indoors, Indoor Growing, Grow Pot, Grow Marijuana.
Agreed, but we have a responsibility to better the information we give out. As a grower, I need to learn what I can to get the best I can.
As someone that gives help to others, I REALLY want the evidence so if I'm wrong, I know why. Knowing why makes it easy to fix, just like knowing how something works. If you know that, you can modify it with ease. Lets avoid Bro Science, evidence please.
 
sambapati

sambapati

1,203
263
Agreed, but we have a responsibility to better the information we give out. As a grower, I need to learn what I can to get the best I can.
As someone that gives help to others, I REALLY want the evidence so if I'm wrong, I know why. Knowing why makes it easy to fix, just like knowing how something works. If you know that, you can modify it with ease. Lets avoid Bro Science, evidence please. OK and fair enough, but first take a look at my grows featuring blue light and a highly controlled air-conditioned environment -- 9/10 females 7/7 females. Other things I've done like using coconut water or germinating in a coconut shell -- that is just me using every possible method to ensure the best result. So, I soak my seeds in specific way and then plant them to ensure success. Where this site doesn't really help end-users -- who 'validates' the validators. I also place black out tape over my extension cord lights.
These things I believe are true:

36 hours of darkness works before flip & B4 chop
All seed manipulation/drying/curing/storage done in complete darkness
But it's gonna take money
A whole lotta spending money
It's gonne take plenty of money
To do it right, child
It's gonna take time
A whole lot of precious time
It's gonna take patience and time

Ayurvedic Narcotic Medicinal Plants - Page 65

books.google.co.id › books



C. R. Karnick · 1996 · ‎Snippet view
FOUND INSIDE – PAGE 65
Furthermore , he observed the effect of boron on the process of sex differentiation . Soaking of the seeds of two hemp varieties in 0.2 and 0.5 percent H , BO3 gave similar results as those obtained with Mn SO4 Herich 162 , 165 noticed ...

Euphytica 140: 95–106, 2004.
C

2004 Kluwer Academic Publishers. Printed in the Netherlands.
95
The sexual differentiation of Cannabis sativa L.: A morphological
and molecular study
V.M. Cristiana Moliterni
1,∗
, Luigi Cattivelli
2
,P.Ranalli
1
& Giuseppe Mandolino
1
1
Istituto Sperimentale per le Colture Industriali, via di Corticella 133, 40128 Bologna, Italy;
2
Istituto Sperimentale
per la Cerealicoltura, via S. Protaso 308, 29017 Fiorenzuola d’Arda, Italy
(

author for correspondence: e-mail: [email protected])
Key words: Cannabis sativa, cDNA-AFLP, dioecy, sex linked markers, sexual differentiation
Summary
Cannabis sativa L. is a dioecious species with sexual dimorphism 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.
Abbreviations: AFLP: Amplified Fragment Length Polymorphism; LINE: Long Interspersed Elements; LTR: Long
Terminal Repeat; SCAR: Sequence Characterized Amplified Region
Introduction
Cannabis sativa belongs to the family of Cannabaceae,
order Rosales (APGII, 2003). It is a naturally dioecious
species with male and female individuals showing
unisexual flowers and characterized by sexual dimor-
phism: male plants are generally taller and slender than
female plants and have a shorter life cycle. Unisexual
flowers are bored in inflorescences that are terminal
at an earlier stage, and terminal or lateral in a later
stage.
Male inflorescence consists of hanging panicles
sometimes branched, generally with few or no leaves
and composed by a variable number of flowers. Male
flower has a perianth of five sepals that encloses the an-
droecium, composed by five stamens bored by subtle
stalks. The anthers at maturity undergo dehiscence lon-
gitudinally, releasing the pollen grains that are mostly
wind dispersed (Mohan Ram and Nath, 1964). Female
inflorescence is a raceme developing at the apex of
the plant or at the axils of leaves or lateral branches.
The female flower has a very simple structure as it


96
is composed by a green bract that completely wraps
the rudimental perianth and the ovary. This latter is
uniloculate and has a short style that distally differen-
tiates a bifid stigma.
The chromosome set of Cannabis sativa is
composed by nine pairs of autosomes and one pair
of sexual chromosomes: X and Y. The male sex is
endowed with an XY pair, and the female one with
an XX pair, similarly to what found in other dioecious
species such as Humulus lupulus, Silene latifolia,
Coccinia indica, Rumex hastatulus;however, sex de-
termination in Cannabis has been supposed to be based
on a X:autosome dosage rather than on an active-Y
mechanism (Westgaard, 1958; Grant et al., 1994).
The Y chromosome in Cannabis is subtelocentric and
characterized by a satellite at the extremity of the short
arm; besides, the long arm is particularly developed
and probably responsible for the difference found be-
tween the male and the female genome sizes (1683 and
1636 Mbp, respectively; Sakamoto et al., 1998). The
X chromosome is submetacentric, and bears a satellite
at the end of short arm. There are no specific reports
about the chromosome set of monoecious plants.
As already reported for many other plant sexual
chromosomes, Cannabis sativa Y chromosome is
strongly heterochromatic and rich of repetitive se-
quences that are likely cause of its marked metaphasic
condensation. A high percent of the repeated DNA
is made of LINE-like sequences (Boecke, 1989),
probably representing traces of transposable elements
showing a low level of transcription for the presence of
still active ORFs, coding for enzymes involved in the
transposition mechanism. Cannabis sativa LINE ele-
ments (LINE-CS) are represented in the X chromosome
and in the autosomes too, but their concentration at the
end of Y chromosome is particularly high. This obser-
vation led to the hypothesis that these sequences might
have a role in maintaining the structure of Y chromo-
some and that they can contribute to the morphological
and structural differentiation of the sex chromosomes,
by creating heteromorphic regions in which the recom-
bination is prevented (Sakamoto et al., 2000; Peil et al.,
2003).
The phenotypic expression of sex in hemp shows
some flexibility. Anomalies in flower development
are sometimes observed, such as the appearance of
hermaphrodite flowers or the development of mixed
inflorescences (bearing both male and female flowers),
like those occurring in the monoecious phenotypes.
Monoecious varieties have been developed from some
of these mutations, and need a strict selection to be
maintained in the variety during the seed multiplica-
tion, due to the recessive nature of the trait.
In some hemp genotypes it is possible to obtain to-
tal or partial reversion of the sex. It is known that the
treatment with masculinizing or feminizing chemical
agents is effective in determining the formation of the
opposite sex reproductive organs even in plants that are
already sexually well differentiated. Chemicals that in-
hibit the biosynthesis or the activity of ethylene, such
as aminoetoxyvinylglycine, silver thiosulphate and sil-
ver nitrate, have a masculinizing effect, while the pre-
cursors or activators of the biosynthesis of ethylene,
like etephon, have a feminizing effect (Mohan Ram
& Sett, 1982a, 1982b). The ability to undergo sexual
reversion is thought to have a genetic base: some eco-
types such as the Italian Carmagnola are very resistant
to any sex reversion treatment, while plants belong-
ing to Fibranova cv. are quite prone to sex reversion
(G. Grassi, E. de Meijer, personal communications).
In Italian open field conditions, the life cycle of a
typical dioecious variety has a 5–6 months duration,
and sexual maturity is attained after 3–4 months, when
the earliest unisexual flowers appear. Sexual dimor-
phism of dioecious hemp is generally apparent only
in a much later stage of development, just before the
onset of flowering, when a marked elongation of the
last internodes occurs in male plants causing them to
become taller and slender than female plants.
Sex is considered an important trait for hemp ge-
netic improvement. The Bredemann’s strategy of se-
lection for fibre quality implies a relatively early quali-
quantitative analysis of fibre in male plants before
pollen dispersion (Bredemann, 1938). This analysis
is followed by the elimination of lower-quality male
plants, not intended for pollination. Therefore, the pos-
sibility of early sex identification and the study of the
mechanism of sexual differentiation in dioecious va-
rieties are of paramount interest in hemp research.
Attempts have been made in the pre-genomic era by
multivariate analysis of morphological traits followed
by correlation to the sex expression (Lacombe, 1980).
Since the Nineties, DNA markers were developed,
capable of discriminating the male plants from the
female and the monoecious ones (Mandolino et al.,
1998, 1999). Such markers (see also the paper by
G. Mandolino and A. Carboni in this special issue) can
be fruitfully used in the selection schemes for hemp
breeding, and in the assessment of the number of male
plants in monoecious seed lots.
Sex linked markers, provided that they are tightly
and reliably associated to the sexual phenotype, can

97
be of great importance in the study of the earliest
stages of Cannabis sexual differentiation. Beyond the
relationship between the presence of the Y chromo-
some and the male sex, very little is known about the
molecular mechanisms underlying sexual differentia-
tion of hemp. We have investigated the onset of sexual
differentiation both at the histological and molecular
level. The characterization of different developmental
stages in male and female plants was carried out by op-
tical microscopy; differentially expressed sequences at
the same developmental stage in the two sexes were
also identified by cDNA-AFLP analysis. The possible
role of these differently expressed sequences in male
and female plants at the onset of sexual differentiation is
discussed.
Materials and methods
Plant material
All the experiments were carried out using cv.
Fibranova, an Italian cross-bred variety, characterized
by high fiber content; Fibranova is dioecious and shows
asex ratio close to 1. Seeds were sown in peat paper
pots, and plants grown in the greenhouse at a 16 h
photoperiod, with a daily thermo period of 11–27

C,
throughout the plant’s life cycle.
Sex identification
Early sex identification was carried out according
to Klimyuk et al. (1993) with minor modifications
(Mandolino and Ranalli, 2002). Tissue fragments
3–4 mm long were picked up from young leaves at
the 2–3

node and collected in sterile 1.5 ml vials;
40 µlofa0.25 M NaOH solution were added to each
vial that was then placed in boiling water (100

C) for
50 s. An amount of 40 µlofa0.25 M NaCl solution
and 20 µlofadetergent-buffer solution (0.5 M Tris
pH 8, 0.25% Triton X-100) were added; the vials
were then centrifuged for few seconds at maximum
speed and placed in boiling water (100

C) for 2 min.
Each tissue fragment was transferred in a PCR vial
and 25 µlofthe PCR reaction mixture for the male
specific marker amplification were added (Mandolino
et al., 1999). Amplicons were run in a 1% agarose gel
in TAE 1X buffer and visualized by U.V. exposition
(305 nm), after ethidium bromide staining. All the
buffers and disposables used in this procedure were
sterile.
Histological methods
Apices from male and female plants grown in the
above described conditions were collected at the
emergence of the leaves of the second, fourth, sixth
and eight nodes. The collected samples were sub-
merged in the fixing solution (glutaraldehyde 3%
in KH
2
PO4/Na
2
PO4 0.1 M buffer, pH 7); this step
sometimes needed a brief treatment in a vacuum jar, to
allow the air to flow out of the stem vessels. Fixing was
obtained after a 1–3 weeks incubation at 4

C. Samples
were then rinsed several times in phosphate buffer
0.1 M, pH 7.0 at 4

C, dehydrated in a graded ethanol
series (ethanol/phosphate buffer, ethanol/water), and
pre-infiltrated in a mixed solution of ethanol and
Technovit 7100 resin (Heraeus Kulzer) for4hat4

C.
The infiltration and embedding steps were carried out
for 36 h at room temperature. Longitudinal sections
2–4 µm thick, obtained with an ultra microtome
(Reichert Jung) fitted with a glass blade, were then
stained with toluidine blue. A toluidine blue 0.1%,
borax 1% aqueous water solution was dropped on the
sections adhering on a glass microscope slide, and was
then dried on a hot plate. Differential staining was ob-
tained by adding few drops of an acid–alcohol solution
(ethanol 70%, HCl 1.5%) and rinsing with water; a drop
of water allowed the cover glass to adhere to the stained
sections that were then observed and photographed
with an optical microscope (Leitz Orthoplan).
RNA extraction and cDNA synthesis
Pooled apices, respectively from undifferentiated
plants at the second node, male plants at fourth node
and female plants at fourth node, were frozen and
grinded under liquid nitrogen. Extraction Buffer
(50 mM Tris-HCl, pH 9.0, 100 mM NaCl, 10 mM
EDTA, 2% SDS) was warmed at 37

C and added
at a 5:1 ratio (v/w) to the powdered sample. Ho-
mogenate was transferred in a sterile polypropylene
tube, extracted twice with an equal volume of a
phenol/chloroform/isoamyl alcohol mixture (25:24:1),
and once with two volumes of a chloroform/isoamyl
alcohol mixture (100:1). Oligo-dT cellulose (Roche)
50 mg/ml and NaCl 0.4 M (final concentrations)
were added to the supernatant. Samples were shaken
(30–50 rpm) for 30 min and then centrifuged for few
minutes at low speed. The cellulose was rinsed twice
with 10 ml of washing Buffer 1 (10 mM Tris-HCl, pH
7.5, 400 mM NaCl, 0.2% SDS) and twice with 10 ml of
washing Buffer 2 (20 mM Tris-HCl, pH 7.5, 100 mM
 
MagicNative1

MagicNative1

109
28
I guess that could be true as far as people who don't do any research about what they are or aren't doin and their experience.. I wouldn't want a newb to decide to try grow and then feel that long after the first pollen sacks emerge they could continue to leave a male in the tent and end up getting their fem plant totally pollinated cuz they weren't smart enough to switch the males clones out progressively as those first sacks are starting to show and then trash them their first few days into flower so there's not a chance to get those results... I just kinda figured it would be something pretty obvious that you wouldn't want to say leave a single male in the tent for half of flower -/+ To attempt something people are clearly talking about as only possibility and even bringing up the topic that I haven't seen anything else that resembles the ""bro science"" I've heard a few times recently

So just in case anyone has been reading I had a plant I thought was fem but started showing male signs anyways.. but the idea was I also had 7 clones that I already knew were male... These were all pre flower male's with no signs at all but my largest plant started to show their first pollen sacks...

My goal was not to push things...
I had a total of 7 more clones of these 3 guaranteed males...
I wanted to take a male and set it in the tent probably no more than 3 days period...
This was merely a plant social experiment and how a fem would react still having juvenile males and possiblities of flowering... Which everyone with any extensive experience would tell you once the fem or males go into flower their main goals at that point aren't necessarily growing anymore it's often doing what's need to have the best possibility of passing on its gene path in the weed world of flowers... It's Chaka Chaka time for these babies...
Some females without a male present at times have been known to turn to transforming themselves into what's known as a Hermie due to stressful situations, but it's also been recorded that fem's will Hermie due to knowing there won't be another chance to pass on anything if they go through this process themselves and self pollinate...

Why, how and all the specifics I could explain somewhat based off research and study but plz don't be fooled I'm no soil scientist and definitely never going to claim to be any type of expert..

But if a female can do these things because of social situations, then my goal was to see if I could find people who Did know for certain not a situation where Someone who thinks it's possible but is definitely mostly bro science because they haven't tried it ever but because they help people would rather no one discusses this...

But they other side of this topic had it went anywhere was also this... What of a fem has been in an environment with multiple males their whole lives and then you stripped them of all possibility to procreate and killed out all the males around... (Probably not much, possible herm situation, but who would know and how did they come about this information... )
Now I actually ended up using that bag seed and magically had all males 🤣 and even had some fun with a few others because those in the know here only days before had told me to trash all 3 of my others because the one I had left was the only fem I was going to have... We'd been watching pretty much daily so we knew what we were looking at... Days later when I had pollen save flowing out the sides of the plant I had pro's telling me how awesome he looked up until it was just undeniable it was male and then true posting again this time by calling him a her and this is where people finally caught on.....

End of story tidbits: first don't just try shit you hear or read and see and don't always trust just anything people on a website tells you just cuz they're the experts here...

But if you do like learning the other ones and outs and want to try experimenting ( cuz after my shit crashed that's all this could have been ), and don't mind even the small possiblity that if your not right and or not ontop of things or doing everything right 🥴 ( that shits to much some times ) then I'd say still don't fuckin do it until you've read watched listened to and studied for a good amount of time and discussed with many...

If you still don't mind these 100% Chances only and bro science mainly type stuff then🤷🏻‍♂️ that's up to you... But you need to know your scale is now on the side of Y being completely screwed and X is only mostly possibly screwed... and try some shit out for fun cuz that's what all this is for anyways past the point of getting some smoke for your personals.. Plus even alot of people in the know here might tell you a 1000 things most might be interesting things that will TRULY help you based off their exp.. but understand even experts here don't know everything... And some don't know certain things that could or aren't true simply because they never wanted to loose out themselves so things the understood or uncertain to them will always be a no as far as they are concerned because they wouldn't know to begin with but choose caution over furthering exp even if that experience is just testing out how screwed a screwed situation can get or to best better the situation based of things that not everyone may or may not know.... Cuz let's be real people been growin legal or illegal for centuries... But to the day we still just don't know everything about or fav plants and how they work on their biological clocks etc because even tho we've all been smoking since we were ✌️ it wasn't that long ago that almost all legit n legal studies into how this plant best works n grows was illegal for anyone including and excluding anyone and everyone not in a certain government contracts and study the ended using to lock down specific patents behind your everyday Americans back...

Moral of the story we all want to better our plants and further our knowledge of the grow, and some are all for learning all the goods and the bads regardless of guaranteed facts... cuz for most of us there was a point where we just didn't know and thanks to everyone else's testing study and knowledge ended up building a awesome community within our communities of your everyday Vipars....

I do hope maybe someone might know a little further into this subject and we do eventually get a badass topic of discussion.... But at the same time it's already mission failure anyways over here... Can't use 30 days of premature males to better a flowering female if that female is a male... 🤣🤣🤣🤣 That stuff was so funny how it worked out to.,. Won't go into any details on this feed tho... It was pretty funny tho one guy was like damn bro you been growing longer than me 🤣🤣🤣🤣
 
MagicNative1

MagicNative1

109
28
These things I believe are true:

36 hours of darkness works before flip & B4 chop
All seed manipulation/drying/curing/storage done in complete darkness
But it's gonna take money
A whole lotta spending money
It's gonne take plenty of money
To do it right, child
It's gonna take time
A whole lot of precious ti
It's gonna take patience and time

Ayurvedic Narcotic Medicinal Plants - Page 65

books.google.co.id › books



C. R. Karnick · 1996 · ‎Snippet view
FOUND INSIDE – PAGE 65
Furthermore , he observed the effect of boron on the process of sex differentiation . Soaking of the seeds of two hemp varieties in 0.2 and 0.5 percent H , BO3 gave similar results as those obtained with Mn SO4 Herich 162 , 165 noticed ...

Euphytica 140: 95–106, 2004.
C

2004 Kluwer Academic Publishers. Printed in the Netherlands.
95
The sexual differentiation of Cannabis sativa L.: A morphological
and molecular study
V.M. Cristiana Moliterni
1,∗
, Luigi Cattivelli
2
,P.Ranalli
1
& Giuseppe Mandolino
1
1
Istituto Sperimentale per le Colture Industriali, via di Corticella 133, 40128 Bologna, Italy;
2
Istituto Sperimentale
per la Cerealicoltura, via S. Protaso 308, 29017 Fiorenzuola d’Arda, Italy
(

author for correspondence: e-mail: [email protected])
Key words: Cannabis sativa, cDNA-AFLP, dioecy, sex linked markers, sexual differentiation
Summary
Cannabis sativa L. is a dioecious species with sexual dimorphism 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.
Abbreviations: AFLP: Amplified Fragment Length Polymorphism; LINE: Long Interspersed Elements; LTR: Long
Terminal Repeat; SCAR: Sequence Characterized Amplified Region
Introduction
Cannabis sativa belongs to the family of Cannabaceae,
order Rosales (APGII, 2003). It is a naturally dioecious
species with male and female individuals showing
unisexual flowers and characterized by sexual dimor-
phism: male plants are generally taller and slender than
female plants and have a shorter life cycle. Unisexual
flowers are bored in inflorescences that are terminal
at an earlier stage, and terminal or lateral in a later
stage.
Male inflorescence consists of hanging panicles
sometimes branched, generally with few or no leaves
and composed by a variable number of flowers. Male
flower has a perianth of five sepals that encloses the an-
droecium, composed by five stamens bored by subtle
stalks. The anthers at maturity undergo dehiscence lon-
gitudinally, releasing the pollen grains that are mostly
wind dispersed (Mohan Ram and Nath, 1964). Female
inflorescence is a raceme developing at the apex of
the plant or at the axils of leaves or lateral branches.
The female flower has a very simple structure as it


96
is composed by a green bract that completely wraps
the rudimental perianth and the ovary. This latter is
uniloculate and has a short style that distally differen-
tiates a bifid stigma.
The chromosome set of Cannabis sativa is
composed by nine pairs of autosomes and one pair
of sexual chromosomes: X and Y. The male sex is
endowed with an XY pair, and the female one with
an XX pair, similarly to what found in other dioecious
species such as Humulus lupulus, Silene latifolia,
Coccinia indica, Rumex hastatulus;however, sex de-
termination in Cannabis has been supposed to be based
on a X:autosome dosage rather than on an active-Y
mechanism (Westgaard, 1958; Grant et al., 1994).
The Y chromosome in Cannabis is subtelocentric and
characterized by a satellite at the extremity of the short
arm; besides, the long arm is particularly developed
and probably responsible for the difference found be-
tween the male and the female genome sizes (1683 and
1636 Mbp, respectively; Sakamoto et al., 1998). The
X chromosome is submetacentric, and bears a satellite
at the end of short arm. There are no specific reports
about the chromosome set of monoecious plants.
As already reported for many other plant sexual
chromosomes, Cannabis sativa Y chromosome is
strongly heterochromatic and rich of repetitive se-
quences that are likely cause of its marked metaphasic
condensation. A high percent of the repeated DNA
is made of LINE-like sequences (Boecke, 1989),
probably representing traces of transposable elements
showing a low level of transcription for the presence of
still active ORFs, coding for enzymes involved in the
transposition mechanism. Cannabis sativa LINE ele-
ments (LINE-CS) are represented in the X chromosome
and in the autosomes too, but their concentration at the
end of Y chromosome is particularly high. This obser-
vation led to the hypothesis that these sequences might
have a role in maintaining the structure of Y chromo-
some and that they can contribute to the morphological
and structural differentiation of the sex chromosomes,
by creating heteromorphic regions in which the recom-
bination is prevented (Sakamoto et al., 2000; Peil et al.,
2003).
The phenotypic expression of sex in hemp shows
some flexibility. Anomalies in flower development
are sometimes observed, such as the appearance of
hermaphrodite flowers or the development of mixed
inflorescences (bearing both male and female flowers),
like those occurring in the monoecious phenotypes.
Monoecious varieties have been developed from some
of these mutations, and need a strict selection to be
maintained in the variety during the seed multiplica-
tion, due to the recessive nature of the trait.
In some hemp genotypes it is possible to obtain to-
tal or partial reversion of the sex. It is known that the
treatment with masculinizing or feminizing chemical
agents is effective in determining the formation of the
opposite sex reproductive organs even in plants that are
already sexually well differentiated. Chemicals that in-
hibit the biosynthesis or the activity of ethylene, such
as aminoetoxyvinylglycine, silver thiosulphate and sil-
ver nitrate, have a masculinizing effect, while the pre-
cursors or activators of the biosynthesis of ethylene,
like etephon, have a feminizing effect (Mohan Ram
& Sett, 1982a, 1982b). The ability to undergo sexual
reversion is thought to have a genetic base: some eco-
types such as the Italian Carmagnola are very resistant
to any sex reversion treatment, while plants belong-
ing to Fibranova cv. are quite prone to sex reversion
(G. Grassi, E. de Meijer, personal communications).
In Italian open field conditions, the life cycle of a
typical dioecious variety has a 5–6 months duration,
and sexual maturity is attained after 3–4 months, when
the earliest unisexual flowers appear. Sexual dimor-
phism of dioecious hemp is generally apparent only
in a much later stage of development, just before the
onset of flowering, when a marked elongation of the
last internodes occurs in male plants causing them to
become taller and slender than female plants.
Sex is considered an important trait for hemp ge-
netic improvement. The Bredemann’s strategy of se-
lection for fibre quality implies a relatively early quali-
quantitative analysis of fibre in male plants before
pollen dispersion (Bredemann, 1938). This analysis
is followed by the elimination of lower-quality male
plants, not intended for pollination. Therefore, the pos-
sibility of early sex identification and the study of the
mechanism of sexual differentiation in dioecious va-
rieties are of paramount interest in hemp research.
Attempts have been made in the pre-genomic era by
multivariate analysis of morphological traits followed
by correlation to the sex expression (Lacombe, 1980).
Since the Nineties, DNA markers were developed,
capable of discriminating the male plants from the
female and the monoecious ones (Mandolino et al.,
1998, 1999). Such markers (see also the paper by
G. Mandolino and A. Carboni in this special issue) can
be fruitfully used in the selection schemes for hemp
breeding, and in the assessment of the number of male
plants in monoecious seed lots.
Sex linked markers, provided that they are tightly
and reliably associated to the sexual phenotype, can

97
be of great importance in the study of the earliest
stages of Cannabis sexual differentiation. Beyond the
relationship between the presence of the Y chromo-
some and the male sex, very little is known about the
molecular mechanisms underlying sexual differentia-
tion of hemp. We have investigated the onset of sexual
differentiation both at the histological and molecular
level. The characterization of different developmental
stages in male and female plants was carried out by op-
tical microscopy; differentially expressed sequences at
the same developmental stage in the two sexes were
also identified by cDNA-AFLP analysis. The possible
role of these differently expressed sequences in male
and female plants at the onset of sexual differentiation is
discussed.
Materials and methods
Plant material
All the experiments were carried out using cv.
Fibranova, an Italian cross-bred variety, characterized
by high fiber content; Fibranova is dioecious and shows
asex ratio close to 1. Seeds were sown in peat paper
pots, and plants grown in the greenhouse at a 16 h
photoperiod, with a daily thermo period of 11–27

C,
throughout the plant’s life cycle.
Sex identification
Early sex identification was carried out according
to Klimyuk et al. (1993) with minor modifications
(Mandolino and Ranalli, 2002). Tissue fragments
3–4 mm long were picked up from young leaves at
the 2–3

node and collected in sterile 1.5 ml vials;
40 µlofa0.25 M NaOH solution were added to each
vial that was then placed in boiling water (100

C) for
50 s. An amount of 40 µlofa0.25 M NaCl solution
and 20 µlofadetergent-buffer solution (0.5 M Tris
pH 8, 0.25% Triton X-100) were added; the vials
were then centrifuged for few seconds at maximum
speed and placed in boiling water (100

C) for 2 min.
Each tissue fragment was transferred in a PCR vial
and 25 µlofthe PCR reaction mixture for the male
specific marker amplification were added (Mandolino
et al., 1999). Amplicons were run in a 1% agarose gel
in TAE 1X buffer and visualized by U.V. exposition
(305 nm), after ethidium bromide staining. All the
buffers and disposables used in this procedure were
sterile.
Histological methods
Apices from male and female plants grown in the
above described conditions were collected at the
emergence of the leaves of the second, fourth, sixth
and eight nodes. The collected samples were sub-
merged in the fixing solution (glutaraldehyde 3%
in KH
2
PO4/Na
2
PO4 0.1 M buffer, pH 7); this step
sometimes needed a brief treatment in a vacuum jar, to
allow the air to flow out of the stem vessels. Fixing was
obtained after a 1–3 weeks incubation at 4

C. Samples
were then rinsed several times in phosphate buffer
0.1 M, pH 7.0 at 4

C, dehydrated in a graded ethanol
series (ethanol/phosphate buffer, ethanol/water), and
pre-infiltrated in a mixed solution of ethanol and
Technovit 7100 resin (Heraeus Kulzer) for4hat4

C.
The infiltration and embedding steps were carried out
for 36 h at room temperature. Longitudinal sections
2–4 µm thick, obtained with an ultra microtome
(Reichert Jung) fitted with a glass blade, were then
stained with toluidine blue. A toluidine blue 0.1%,
borax 1% aqueous water solution was dropped on the
sections adhering on a glass microscope slide, and was
then dried on a hot plate. Differential staining was ob-
tained by adding few drops of an acid–alcohol solution
(ethanol 70%, HCl 1.5%) and rinsing with water; a drop
of water allowed the cover glass to adhere to the stained
sections that were then observed and photographed
with an optical microscope (Leitz Orthoplan).
RNA extraction and cDNA synthesis
Pooled apices, respectively from undifferentiated
plants at the second node, male plants at fourth node
and female plants at fourth node, were frozen and
grinded under liquid nitrogen. Extraction Buffer
(50 mM Tris-HCl, pH 9.0, 100 mM NaCl, 10 mM
EDTA, 2% SDS) was warmed at 37

C and added
at a 5:1 ratio (v/w) to the powdered sample. Ho-
mogenate was transferred in a sterile polypropylene
tube, extracted twice with an equal volume of a
phenol/chloroform/isoamyl alcohol mixture (25:24:1),
and once with two volumes of a chloroform/isoamyl
alcohol mixture (100:1). Oligo-dT cellulose (Roche)
50 mg/ml and NaCl 0.4 M (final concentrations)
were added to the supernatant. Samples were shaken
(30–50 rpm) for 30 min and then centrifuged for few
minutes at low speed. The cellulose was rinsed twice
with 10 ml of washing Buffer 1 (10 mM Tris-HCl, pH
7.5, 400 mM NaCl, 0.2% SDS) and twice with 10 ml of
washing Buffer 2 (20 mM Tris-HCl, pH 7.5, 100 mM
Yeah that was alright... Mostly talks about the differences in male female Xx & Xy.... I didn't yet read all of that probably will tomorrow on my way up to Oklahoma a bit more... I did scan through n saw they were explaining some testing etc... Some as far back as 1938.. which is cool...

One thing I didn't see in there that would have been interesting since they already were touching the topic is on polyploids...

Polyploid cannabis is a chromosome-related variation of normal cannabis. ... But if you have three or more sets of chromosomes (instead of two) the organism is said to exhibit polyploid characteristics. Polyploid cannabis would have 3, or more, paired chromosomes instead of the normal two.Sep 6, 2019

This has created an offshoot in polymorphism 🤣
 
ComfortablyNumb

ComfortablyNumb

5,770
313
Ok, cool. Thanx for sharing that.
So, this study says that the sex of the plant is changeable until sometime as early as the 4th node. After that, it requires a stressor to initiate the change. STS is mentioned because it chemically changes the bonds that hold the chromosomes 'in place' and they are influenced heavily toward being male. Very interesting.

So, the seed has both male and female in it and it depends not strictly on environment, but specific things in the environment that can affect the chromosomal bond.
Cool. Thank you for sharing.
 

Similar threads

I
Replies
23
Views
984
N1ghtL1ght
N1ghtL1ght
MagicNative1
Replies
8
Views
326
MagicNative1
MagicNative1
Dudethatsweet
Replies
2
Views
132
josefrahl
josefrahl
MagicNative1
Replies
0
Views
58
MagicNative1
MagicNative1
Top Bottom