Clone of a Clone of a... Degredation Experiment

[Snype]

Comparing tolerance to degradation is just plain retarded.

 

[Regulator Dave]

Epigenics

Epigenics

I found this article @ http://en.wikipedia.org/wiki/Epigenetics

"In biology, and specifically genetics, epigenetics is the study of inherited changes in phenotype (appearance) or gene expressionDNAepi- (Greek: επί- over, above) -genetics.

These changes may remain through cell divisions for the remainder of the cell's life and may also last for multiple generations.

However, there is no change in the underlying DNA sequence of the organism;[1] instead, non-genetic factors cause the organism's genes to behave (or "express themselves") differently.[2] caused by mechanisms other than changes in the underlying sequence, hence the name"

Perhaps what we should identify and look at closer are epigenetic changes? Is it possible or a contradiction to think some epigenetic changes could be "permanent" in that they are carried from clone to clone but not permanent in the sense that if you breed with those clones the traits would not be passed to the progeny?


I am not making a assertion with a question, I am merely asking a question here.

 

[Grat3fulh3ad]

I found this article @ http://en.wikipedia.org/wiki/Epigenetics

"In biology, and specifically genetics, epigenetics is the study of inherited changes in phenotype (appearance) or gene expressionDNAepi- (Greek: επί- over, above) -genetics.

These changes may remain through cell divisions for the remainder of the cell's life and may also last for multiple generations.

However, there is no change in the underlying DNA sequence of the organism;[1] instead, non-genetic factors cause the organism's genes to behave (or "express themselves") differently.[2] caused by mechanisms other than changes in the underlying sequence, hence the name"

Perhaps what we should identify and look at closer are epigenetic changes? Is it possible or a contradiction to think some epigenetic changes could be "permanent" in that they are carried from clone to clone but not permanent in the sense that if you breed with those clones the traits would not be passed to the progeny?


I am not making a assertion with a question, I am merely asking a question here.

epigenetic changes are heritable, but are also temporary... heritable just means that it can last through multiple cell divisions, not that it is permanent.

 

[VerdantGreen]

hey head i thought that epigenetics was ony applicable to breeding. can cuttings/clones also have heritable epigenetic changes???

 

[Grat3fulh3ad]

hey head i thought that epigenetics was ony applicable to breeding. can cuttings/clones also have heritable epigenetic changes???

epigenetics are not really applicable to breeding, except that unhealthy plants make substandard seeds which are harder to get going.

It is only applicable to clones if the conditions which caused the changes persist.

heritable changes does not necessarily inherited to the next generation of organism, and epigenetic changes are usually only heritable in that they last through multiple cell divisions within an organism

 

[VerdantGreen]

epigenetics are not really applicable to breeding, except that unhealthy plants make substandard seeds which are harder to get going.

It is only applicable to clones if the conditions which caused the changes persist.

heritable changes does not necessarily inherited to the next generation of organism, and epigenetic changes are usually only heritable in that they last through multiple cell divisions within an organism

thanks head thats pretty much how i thought it was - except the bit about clones...
surely the environmental conditions causing change in clones would be a change in phenotype??

 

[headband 707]

^^^ what Head said" .^^^ lol

 

[Grat3fulh3ad]

thanks head thats pretty much how i thought it was - except the bit about clones...
surely the environmental conditions causing change in clones would be a change in phenotype??

Phenotype simply means "the observable characteristics of an individual resulting from the interaction of its genotype with the environment."

All physical characteristics are phenotype.

So based on that... Yes, any change in any physical characteristic is a change in phenotype.

In other words epigenetic changes can cause changes in phenotype, but also might not... and there are many factors aside from epigenetic change which can cause changes in phenotype.

 

[Microbeman]

We do not understand the underlying cause of Alzheimers, so I'm not exactly sure what you mean by this...

Sorry for bringing in current research findings which are yet to be hard theory. It is relatively new research that injury to chromosomes [eg. degradation of telomeres] is likely the cause of many contemporary maladies, especially neurological ones such as Alzheimers. However one may logically hypothesis utilizing available statistics and data (evidence of DNA deformity – samples taken) that many of the diseases which are on the rise are very likely resultant from environmentally caused DNA damage.

 

[Microbeman]

I don’t know if any DNA/genetic testing was done here. Perhaps someone with journal access could follow up. There may be citations or leads within this paper which could help.

http://www.ingentaconnect.com/content/saf/fs/1989/00000035/00000004/art00004#expand/collapse

 

[Regulator Dave]

Molecular Basis of Epigenics

Molecular Basis of Epigenics

I found this information @ http://en.wikipedia.org/wiki/Epigenetic_inheritance

Uhg, it is another wikipedia link

"The molecular basis of epigenetics is complex. It involves modifications of the activation of certain genes, but not the basic structure of DNA. Additionally, the chromatin proteins associated with DNA may be activated or silenced. This accounts for why the differentiated cells in a multi-cellular organism express only the genes that are necessary for their own activity. Epigenetic changes are preserved when cells divide. Most epigenetic changes only occur within the course of one individual organism's lifetime, but, if a mutation in the DNA has been caused in sperm or egg cell that results in fertilization, then some epigenetic changes are inherited from one generation to the next.[9] This raises the question of whether or not epigenetic changes in an organism can alter the basic structure of its DNA (see Evolution, below), a form of Lamarckism."

"Epigenetic changes are preserved when cells divide"

If epigenetic changes are preserved when cells divide, is it not plausible that a Cannabis plant could undergo an epigenetic change that would be passed on from cell to cell as the plant grew? Thus changing the cells of the limbs from which the cuttings are taken, therefor passing the change to the clone and that clone growing with each cell passing the epigenetic change carrying the change into the clone line?

"Most epigenetic changes only occur within the course of one individual organism's lifetime," If a plant from seed is cloned and that clone is cloned and this is done for 20 years, Is the 20 year old clone NOT the same plant as the first plant?

An epigenetic change could be viewed by us as positive or negative but a change none the less.

This is of coarse noting that no change in the DNA sequence ever took place as it would no longer be considered epigenetic.

Don't get me wrong I am just asking questions concerning what could happen and am not making claims I have observed it happen or am trying to use this info to explain why some people feel like they have observed clone degradation.

 

[Mr.Jones]

in epigenetics as far as i know there are just methyl side chains added to the aminoacids, which activate or inactivate certain parts of the dna (promoter activation and so on).
this part of biology is yet to be discovered but as stated before the epigenetic change can hold up for some cell generations - take a cutting and see how many cells there are at the beginning and how many there are after flower - this epigenetic change must be a big factor if it would keep itself up for so many generations.
i bet nobody argues about the difficulties in early development stages of clones which were taken from a sick mother. one thing we have to keep in mind is that a plant is a organism of constant growth and the human beeing is not! so if a human cell is in a epigeneticly "bad mood" this could hold on forever - but looking at cannabis sativa and the possiblilty to control vegetative growth by the lightphases there is no need to worry about epigenetics. maybe one run is going to be worse than the others but well ...

also epigenetics would be much more an argument speaking for taking clones from clones since there is much more cell production and many new formed cells.

Mr.Jones

We do not understand the underlying cause of Alzheimers

as far as im informed the actuall cause of Alzheimer is the incorrect folding of Proteins - also causes BSE ... maybe resulting from a defect before ...

 

[Grat3fulh3ad]

I found this information @ http://en.wikipedia.org/wiki/Epigenetic_inheritance

Uhg, it is another wikipedia link

"The molecular basis of epigenetics is complex. It involves modifications of the activation of certain genes, but not the basic structure of DNA. Additionally, the chromatin proteins associated with DNA may be activated or silenced. This accounts for why the differentiated cells in a multi-cellular organism express only the genes that are necessary for their own activity. Epigenetic changes are preserved when cells divide. Most epigenetic changes only occur within the course of one individual organism's lifetime, but, if a mutation in the DNA has been caused in sperm or egg cell that results in fertilization, then some epigenetic changes are inherited from one generation to the next.[9] This raises the question of whether or not epigenetic changes in an organism can alter the basic structure of its DNA (see Evolution, below), a form of Lamarckism."

"Epigenetic changes are preserved when cells divide"

If epigenetic changes are preserved when cells divide, is it not plausible that a Cannabis plant could undergo an epigenetic change that would be passed on from cell to cell as the plant grew? Thus changing the cells of the limbs from which the cuttings are taken, therefor passing the change to the clone and that clone growing with each cell passing the epigenetic change carrying the change into the clone line?

"Most epigenetic changes only occur within the course of one individual organism's lifetime," If a plant from seed is cloned and that clone is cloned and this is done for 20 years, Is the 20 year old clone NOT the same plant as the first plant?

An epigenetic change could be viewed by us as positive or negative but a change none the less.

This is of coarse noting that no change in the DNA sequence ever took place as it would no longer be considered epigenetic.

Don't get me wrong I am just asking questions concerning what could happen and am not making claims I have observed it happen or am trying to use this info to explain why some people feel like they have observed clone degradation.

The short answer to your question is : not very likely.

the most common epigenetic changes are those that make an undifferentiated cell become a specific type of cell. That is why, even though every cell in your body has the same DNA, some cells grow are muscle cells, and some liver cells, and some bone cells, and some brain cells. Same DNA but different portions turned on or off.


Epigenetic changes are not going to turn a short person into a tall person.

it's mostly a local cellular level thing, not an organism wide thing.

 

[beta]

Sorry for bringing in current research findings which are yet to be hard theory. It is relatively new research that injury to chromosomes [eg. degradation of telomeres] is likely the cause of many contemporary maladies, especially neurological ones such as Alzheimers. However one may logically hypothesis utilizing available statistics and data (evidence of DNA deformity – samples taken) that many of the diseases which are on the rise are very likely resultant from environmentally caused DNA damage.

Sounds like very interesting research; I have no doubt that it'll have a serious impact on this discussion once all of the information is in...

 

[Microbeman]

Sounds like very interesting research; I have no doubt that it'll have a serious impact on this discussion once all of the information is in...

Well I doubt it will have much to do with this topic. It was only meant to illustrate environmentally caused chromosomal damage. I believe this is actually quite well established.

 

[Microbeman]

This is ‘only’ to help establish that chromosomal damage can occurr in plants, seeds, cuttings

http://www.inchem.org/documents/ehc/ehc/ehc51.htm#SectionNumber:2.2

World Health Orgnization Geneva, 1985


2.6. The Use of Higher Plants to Detect Mutagenic Chemicals

“2.6.1. Introduction Many of the fundamental concepts of modern genetics were established in higher plants and the term "mutation" was introduced by the Dutch botanist, Hugo de Vries, in 1909, to describe a sudden hereditary change in Oenothera lamarckiana. Plant systems played a major part in early investigations of the genetic changes caused by radiation (Read, 1959; Revell, 1959) and a variety of plants have been used to study the mutagenic effects of chemicals at the gene and chromosome levels. With the increasing concern over the genotoxicity of sophisticated techniques for studying mutations in bacteria, lower plants, insects, and mammalian cells, there has been a loss of interest in the testing of potentially mutagenic chemicals in higher plant systems. This is surprising as plants appear to offer significant advantages over other organisms in certain circumstances, though they have, of course, important limitations. Techniques for studying mutagenic chemicals have been developed in about 10 species of higher plants and a whole range of specific genetic end-points are available. Mitotic chromosome alterations can be studied in the somatic cells from root tips, or pollen tubes in, for example, barley, the broad bean, or the onion. Pollen mother cells from a number of species are suitable for detecting chemically-induced chromosomal aberrations in meiotic cells. Gene mutations at specific loci can be investigated in maize or soybean plants and multilocus mutation systems are available in barley and maize. The chromosome systems allow the observation of structural chromosome damage and effects on chromosome segregation and general mitotic function. The chromosomes are morphologically similar, and appear to respond to treatment with mutagens in a similar way to those of mammals and other eukaryotes. A survey of the literature prepared under the US Environmental Protection Agency Gene-Tox Program (Constantin & Owens, 1982) revealed that about 350 compounds, covering a wide range of chemical classes, had been tested for mutagenic activity in plants. The same authors also compared the results of testing eight model mutagens in plants with the results obtained in other systems. They claimed that the correlation between plant data and results from cultured mammalian cells was at least as good as that with data derived from bacteria and Drosophila. A comparison of the results of testing a series of pesticides in plant root tips and mammalian cells for chromosomal aberrations showed a remarkable qualitative similarity between the two sets of results. However, the data on chromosome damage in mammalian cells for some of the pesticides was not truly representative of the literature on these chemicals. Although a database representing more than 350 compounds tested in plant systems has been assembled, a large proportion of the chemicals tested were shown to be mutagenic in one plant system or another, and there is a significant lack of information on non-mutagenic chemicals. “


“(b) Tradescantia paludosa Strains of T. paludosa that proliferate and propagate easily and quickly under local environmental conditions should be used. A suitable clone should grow to maturity from cuttings in 40 - 60 days. Since the chromosomes of pollen mother cells are not of adequate quality for the detailed analysis of metaphase aberrations, a technique has been developed for detecting chromosome breakage on the basis of micronuclei at the tetrad stage. In practice, the inflorescences are removed from the plant and the stems placed in solutions of the test chemical. Alternatively, the buds can be exposed to gaseous materials in a suitable chamber. The optimum length of treatment is determined experimentally and a recovery period of 24 - 30 h is necessary to allow chromosome damage in early prophase 1 to reach the tetrad stage where micronuclei can be scored. Micronuclei are assumed to be a result of either chromosome fragmentation or of whole chromosomes lost during meiosis and are therefore a measure of both structural damage and aneuploidy (or non-disjunction). It is usual to score between 1000 and 1500 tetrads from each experimental group including both negative and positive controls. “

Of course this was pre-Monsanto and these days maize and soy might be impervious to chromosome damage. Hahaha > little science humour.

 

[jeffie]

What is your point, exactly? What on earth has quantum physics got to do with the question/hypothesis originally posted (which, incidentally, has been put to bed already). Are you just trying to make yourself feel intelligent by letting us all know you've heard of Einstein or Bohr? Well done, mission accomplished, we're all suitably surprised.
This forum is for botanical topics- hence the title. Probably best to showcase your superior knowledge of quantum mechanics at the LHC website, I'm sure they're all much more on your wavelength/frequency of vibration.

Take it easy don't be so hard on yourself or else you're going to mutate and your offspring won't have much vigor. That's at least a possibility, say, due to stress in anger, innit.

To answer your question: my point was that science is largely driven by questioning previous "scientifically sound" theories and by challenging predecessors' scientists' work. But more than that my point "exactly" was that science is always relative and 303hydro posted a good link for us to follow up if interested http://www.youtube.com/watch?v=tevWzkVrk2s There was no need to call him names or how'd you know he won't stress. It seems to me that any belief system is enemy to science and still science is totally dependent on beliefs and how we see the world.

Please, don't feel offended by the quantum theory analogy if the field terrifies you too much. I know it terrifies me if that helps. I feel honored I have impressed You-All so suitably surprised. Thank you.

Now, how does it relate to Cannabis cloning (or cutting and rooting) - we need to think outside the box here. Obviously some of us think they have experienced loss in vigor due to cloning clones and others feel they've missed on this and their cuts always stayed the same. And none run one old mother alongside let alone trying to quantify. So outside the box it must be, imo.

 

[Grat3fulh3ad]

right on... but nobody said chromosomal damage is impossible, just that it is very rare and not likely to be caused by the process or perpetual cloning.

indoor monoculture gardens are very susceptible to pathogen.

infected plants can absolutely give infected cuts, and it can easily worsen over time.

Every single thing that people anecdotally offer as evidence of genetic drift from perpetual cloning can be better explained by pathogen.

 

[Sam_Skunkman]

Anyone have studies that show damage from clones of clones of clones?
Here are a few papers all saying they can't find damage from cloning:


http://home.olemiss.edu/~suman/Geneticstability.pdf

Assessment of the Genetic Stability of Micropropagated Plants of Cannabis sativa by ISSR Markers.
Lata H, Chandra S, Techen N, Khan IA, Elsohly MA
National Center for Natural Products Research, School of Pharmacy, The University of Mississippi, University, MS, USA.
Planta Med 2009 Jul 27.
Inter-simple sequence repeat (ISSR) markers were used to evaluate the genetic stability of the micropropagated plants of CANNABIS SATIVA over 30 passages in culture and hardening in soil for 8 months. A total of 15 ISSR primers resulted in 115 distinct and reproducible bands. All the ISSR profiles from micropropagated plants were monomorphic and comparable to mother plants, confirming the genetic stability among clones and mother plants. Chemical analysis of cannabinoids, using gas chromatography/flame ionization detection (GC/FID), was done to further confirm whether the qualitative and quantitative differences in the major secondary metabolites exist between the mother plant and micropropagated plants. Six major cannabinoids - Delta(9)-THC, THCV, CBD, CBC, CBG, and CBN - were identified and compared with the mother plant. Our results clearly showed a similar cannabinoid profile and insignificant differences in THC content between the two types of plants. These results suggest that the micropropagation protocol developed by us for rapid IN VITRO multiplication is appropriate and applicable for clonal mass propagation of C. SATIVA.

https://www.thieme-connect.de/ejournals/abstract/plantamedica/doi/10.1055/s-0029-1240628



Biochemistry, Molecular Biology and Biotechnology
Original Papers Planta Med 2010; 76(7): 743-750
DOI: 10.1055/s-0029-1240628

© Georg Thieme Verlag KG Stuttgart · New York



Assessment of Cannabinoids Content in Micropropagated Plants of Cannabis sativa and Their Comparison with Conventionally Propagated Plants and Mother Plant during Developmental Stages of Growth

Suman Chandra1, Hemant Lata1, Zlatko Mehmedic1, Ikhlas A. Khan1,2, Mahmoud A. ElSohly1,3
1 National Center for Natural Product Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, University, MS, USA
2 Department of Pharmacognosy, School of Pharmacy, University of Mississippi, University, MS, USA
3 Department of Pharmaceutics, School of Pharmacy, University of Mississippi, University, MS, USA
Abstract

Gas chromatography-flame ionization detection (GC‐FID) was used to assess the chemical profile and quantification of cannabinoids to identify the differences, if existing, in the chemical constituents of in vitro propagated plants (IVP), conventionally grown plants (VP) and indoor grown mother plants (MP-Indoor) of a high THC yielding variety of Cannabis sativa L. during different developmental stages of growth. In general, THC content in all groups increased with plant age up to a highest level during the budding stage where the THC content reached a plateau before the onset of senescence. The pattern of changes observed in the concentration of other cannabinoids content with plants age has followed a similar trend in all groups of plants. Qualitatively, cannabinoids profiles obtained using GC‐FID, in MP-indoor, VP and IVP plants were found to be similar to each other and to that of the field grown mother plant (MP field) of C. sativa. Minor differences observed in cannabinoids concentration within and among the groups were not found to be statistically significant. Our results confirm the clonal fidelity of IVP plants of C. sativa and suggest that the biochemical mechanism used in this study to produce the micropropagated plants does not affect the metabolic content and can be used for the mass propagation of true to type plants of this species for commercial pharmaceutical use.

Key words

Cannabis sativa - Cannabaceae - cannabinoids - Δ9‐tetrahydrocannabinol - gas chromatography‐flame ionization detection - micropropagation


https://www.thieme-connect.de/ejournals/abstract/plantamedica/doi/10.1055/s-0030-1249773


Biochemistry, Molecular Biology and Biotechnology
Original Papers Planta Med 2010; 76(14): 1629-1633
DOI: 10.1055/s-0030-1249773

© Georg Thieme Verlag KG Stuttgart · New York



High Frequency Plant Regeneration from Leaf Derived Callus of High Δ9-Tetrahydrocannabinol Yielding Cannabis sativa L.

Hemant Lata1, Suman Chandra1, Ikhlas A. Khan1,2, Mahmoud A. ElSohly1,3
1 National Center for Natural Products Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi, University, MS, USA
2 Department of Pharmacognosy, School of Pharmacy, University of Mississippi, University, MS, USA
3 Department of Pharmaceutics, School of Pharmacy, University of Mississippi, University, MS, USA
Abstract

An efficient in vitro propagation protocol for rapidly producing Cannabis sativa plantlets from young leaf tissue was developed. Using gas chromatography-flame ionization detection (GC‐FID), high THC yielding elite female clone of a drug-type Cannabis variety (MX) was screened and its vegetatively propagated clones were used for micropropagation. Calli were induced from leaf explant on Murashige and Skoog medium supplemented with different concentrations (0.5, 1.0, 1.5, and 2.0 µM) of indole- 3-acetic acid (IAA), indole- 3- butyric acid (IBA), naphthalene acetic acid (NAA), and 2,4-dichlorophenoxy-acetic acid (2,4-D) in combination with 1.0 µM of thidiazuron (TDZ) for the production of callus. The optimum callus growth and maintenance was in 0.5 µM NAA plus 1.0 µM TDZ. The two-month-old calli were subcultured to MS media containing different concentrations of cytokinins (BAP, KN, TDZ). The rate of shoot induction and proliferation was highest in 0.5 µM TDZ. Of the various auxins (IAA, IBA, and NAA) tested, regenerated shoots rooted best on half strength MS medium (1/2 - MS) supplemented with 2.5 µM IBA. The rooted plantlets were successfully established in soil and grown to maturity with no gross variations in morphology and cannabinoids content at a survival rate of 95 % in the indoor growroom.

Key words

Cannabis sativa - Cannabaceae - callus induction - Δ9‐tetrahydrocannabinol - GC‐FID - organogenesis

Here is another paper I find interesting, (SCAR) markers were used to ID males and females prior to flowering:

https://www.thieme-connect.de/ejournals/abstract/plantamedica/doi/10.1055/s-0030-1249978

 

[GreenintheThumb]

Thread over!