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NewScientist
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Methods for Fruiting Isolates
#10465704 - 06/07/09 12:34 PM (14 years, 8 months ago) |
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I understand that fruiting out each isolated strain is necessary when looking for particular characteristics.
However, can you please share your methods for fruiting the isolates?
For example, how big of a grow do you need to test each isolate? Do I need to fruit out an entire tray of bulk substrate for each isolate, or can I use a smaller amount - say a half-pint size of bulk for each one.
What I'm thinking is I have a butt-load of half-pint jars leftover from my PF tek days. I'll mix up a big batch of horse poo substrate then spawn four half-pint jars with each isolated strain trying to keep the exact same spawn rate for each. That way each strain gets a consistently formulated bulk substrate. And then all the jars can be introduced to the fruiting chamber at the same time to provide consistent fruiting conditions. What I'm trying to do is control for as many variables as possible so that the only thing that is different is the strain.
If you have other or better ways of fruiting and comparing isolated strains, please let me know. Thanks a lot.
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ScavengerType


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Re: Methods for Fruiting Isolates [Re: NewScientist]
#10465763 - 06/07/09 12:48 PM (14 years, 8 months ago) |
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I have heard that doing just BRF cakes will show potency of a particular strain the easiest and the best, it doesn't mater what the substrate of your cake is as a strain will produce equal potency on another one as long as nutrients allow. This is why BRF is recommended for this, because it will not cause any nutrient deficiencies to pollute the results.
-------------------- "Have you ever seen what happens when a grenade goes off in a school? Do you really know what you’re doing when you order shock and awe? Are you prepared to kneel beside a dying soldier and tell him why he went to Iraq, or why he went to any war?" "The things that are done in the name of the shareholder are, to me, as terrifying as the things that are done—dare I say it—in the name of God. Montesquieu said, "There have never been so many civil wars as in the Kingdom of God." And I begin to feel that’s true. The shareholder is the excuse for everything." - Author and former M6/M5 agent John le Carré on Democracy Now. Conquer's Club
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Livingston
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Hi,
I'm not sure if this is possible with P.cubensis, but I plan to try it...why not fruit off the agar in the petri dish first?
That way you will know it's a fruiting isolate and you won't waste time waiting to colonize, spawn and fruit to find out if it's non-fruiting isolate or not.
Then with known fruiters create cakes like the previous poster said. However, I would suggest you test on the substrate you intend to use. If you doing bulk then test on bulk when choosing an 'elite' isolate. If fruiting from cakes then test with cakes...IMO you want to try to replicate the fruiting environment as close as you can so you get a good idea of how they preform in your growing style.
-------------------- Peace and Pasta
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potatonet
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Re: Methods for Fruiting Isolates [Re: Livingston]
#10465846 - 06/07/09 01:09 PM (14 years, 8 months ago) |
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I would suggest doing a BRF cake,
I did a whole tray, thankfully my isolate turned out to be awesome so Im pleased
-------------------- In the Quechua language of Peru there are over 1000 words for "potato"
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veda_sticks
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Re: Methods for Fruiting Isolates [Re: Livingston]
#10465852 - 06/07/09 01:10 PM (14 years, 8 months ago) |
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Im sure i have heard of people letting agar pin to make sure they have a fruiting capable isolate.
PF cakes are fine. it wills how that the strian will fruit, how well it will fruit and to a certain extent how large the fruits will get. Size of fruits will however depend on pinset particularly with cakes since they dont hold much moisture, if you get a stellar pinset on cakes, there going to be smaller mushrooms. Ive seen quite a few cakes covered complelty in mushrooms they were small but the overall amount made up for the lack of size.
Ive heard of some people still using cakes as testers for there strains
-------------------- PF TEK - writeup by EvilMushroom666 Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD Cakes can and will pin! - So you think cakes suck for pins. Your wrong Franks Simple Coir/Verm Tek Franks Proper Pasturisation Tek Franks Spawning To Bulk - Monotub Professor Pinheads RTV Injection Port Tek Foo Mans No Soak WBS Prep Tek
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Livingston
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Re: Methods for Fruiting Isolates [Re: veda_sticks]
#10465888 - 06/07/09 01:18 PM (14 years, 8 months ago) |
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Hey,
Yea I thought I've seen pics of P.cubensis fruiting in my travels...thanks. 
If testing only for potency than sure, cakes would be fine. But if testing to find an elite isolate for a particular environment then test fruiting within that environment would be best IMO. A shotgun tub and Martha have different micro-environments (humidity, temperature, etc), those can affect the way the fruit body develops, etc. I don't think it's required to test fruit in the same environment one will use when growing...but I think it's wise to get a more accurate representation of how each isolate will grow.
-------------------- Peace and Pasta
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NewScientist
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Re: Methods for Fruiting Isolates [Re: Livingston]
#10465944 - 06/07/09 01:28 PM (14 years, 8 months ago) |
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I wouldn't want to us BRF cakes to test the isolates because I always grow on a horse poo and I want a candidate that performs well on that particular substrate. Also, I'm not very excited to go from agar to syringe - how would you do that anyway? I'd use the agar from each isolate to inoculate grain masters, then spawn to horse poo substrate packed into half-pint jars covered in foil - then "birthed" and fruited in a chamber when fully colonized.
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ScavengerType


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Re: Methods for Fruiting Isolates [Re: NewScientist]
#10466011 - 06/07/09 01:43 PM (14 years, 8 months ago) |
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for my jars all I ever did was cut a wedge and toss it into the jars. Just be quick about it and it should be fine.
-------------------- "Have you ever seen what happens when a grenade goes off in a school? Do you really know what you’re doing when you order shock and awe? Are you prepared to kneel beside a dying soldier and tell him why he went to Iraq, or why he went to any war?" "The things that are done in the name of the shareholder are, to me, as terrifying as the things that are done—dare I say it—in the name of God. Montesquieu said, "There have never been so many civil wars as in the Kingdom of God." And I begin to feel that’s true. The shareholder is the excuse for everything." - Author and former M6/M5 agent John le Carré on Democracy Now. Conquer's Club
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potatonet
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you take a piece of agar and make an LC with it or just drop the pieces of agar onto your spawn in the jar, I did WBS spawn with a slice of agar each, next time I will do an LC of the isolate as it will go about 10X faster
-------------------- In the Quechua language of Peru there are over 1000 words for "potato"
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RogerRabbit
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Re: Methods for Fruiting Isolates [Re: potatonet]
#10468360 - 06/07/09 10:32 PM (14 years, 8 months ago) |
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I use brf cakes to test isolates. What leads you to think if you're going to grow on horse manure later that you need to test your isolates on horse manure? You can find the best isolates on brf cakes and then grow them on horse manure later for larger yields. You'll hear all sorts of stories about how you need to test on the substrate you'll use later, and there might be some truth to that with the harder to grow edibles, but cubes will grow on anything, so it's not applicable. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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potatonet
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Re: Methods for Fruiting Isolates [Re: RogerRabbit]
#10469176 - 06/08/09 01:51 AM (14 years, 8 months ago) |
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another quick question on this isolation thing...
I picked an Isolate that did pretty well on my last trays,
however I left the plate that I took the isolate off of and one of the other fans fruited in vitro.
should I take a clone of that fruit? Cause its pretty big for an in vitro fruit.
better strains maybe fruit in vitro?
-------------------- In the Quechua language of Peru there are over 1000 words for "potato"
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Livingston
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Re: Methods for Fruiting Isolates [Re: RogerRabbit]
#10470325 - 06/08/09 11:22 AM (14 years, 8 months ago) |
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Hi
To me it's just common sense: test in the environment where you will grow. To have the most accurate representation of how an organism will preform in a given environment one should use that environment when testing.
I have found that some phenos will preform better than others in different fruiting environments, not in regards to potency necessarily, but in terms of colonization speed, fruit body development/expansion, etc. This is a common trait in most all organisms of the five Kingdoms, and is how 'elite' phenos (ie. sub-strains) are found in mycology, horticulture, agriculture, microbology, etc.
-------------------- Peace and Pasta
Edited by Livingston (06/08/09 11:35 AM)
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RogerRabbit
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Re: Methods for Fruiting Isolates [Re: Livingston]
#10470432 - 06/08/09 11:43 AM (14 years, 8 months ago) |
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Organisms and species, yes, but not 'strains' of a single species, which have the same growth requirements. After 35 years of growing all sorts of mushrooms, I haven't seen one single shred of evidence to support the argument that 'this or that strain' does better on 'this or that substrate'. I've often heard that mantra repeated, and even read it in books, but haven't seen it to be so, based on experience. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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Livingston
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Re: Methods for Fruiting Isolates [Re: RogerRabbit]
#10470773 - 06/08/09 12:42 PM (14 years, 8 months ago) |
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Hi,
Well I am talking about phenotypes within each strain. That is why the same environment used for growth is also used for testing (to limit the variables). For example within SA one will find different phenotypes which react differently to each given environment. To find the 'elite' phenotype it is most common in all fields of science to use the same environment for growth as for testing.
Within the species of P.cubensis there are many stains, those identified by area of origin (ie. land-race genotypes). For example SA and Ecuador are different strains and within their population they will have different phenotypes which perform differently in different environments.
You have much more experience than I do so I value your opinion. It could be that P.cubensis phenotypes within each strain are not very variable and that would make sense because a land-race genotype will be fairly humongous in terms of fruit body phenotypes.
Thanks bro, I always like reading your opinion
-------------------- Peace and Pasta
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Livingston
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Re: Methods for Fruiting Isolates [Re: Livingston]
#10470786 - 06/08/09 12:45 PM (14 years, 8 months ago) |
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Hi
Here is something I wrote yesterday in the strain thread, but I think it's applicable here too: ---------------------
Quote:
apoonanor said: The term "Strain" when describing cubensis probably comes from weed. Weed DOES come in different strains, so the term was used to market shrooms to hippies familiar with the lingo.
I disagree. A strain in terms of intraspecies terminology is a different land-race. That is, P.cubensis from Amazon will be a different strain than one from Ecuador (for example). They will not have the same geno-type, or pheno-type, so they are different intraspecies strains...the same goes for Cannabis spp. with caveats (see below).
If one was to breed say a PE and SA together they would have a intraspecies cross. If one was to breed two different species of Psilocybe spp. (ex. P.cubensis x P.azurescens) that person would be making a intergenus hybrid. A interspecies cross could be called a strain once it reaches the F4-F6 stage and is pretty homogeneous phenotypes.
A strain title is a designation for that geno-type/area of origin, not all P.cubensis are the same strain, though they are the same species.
Humans are the same species but black and white people are different races (eg. strains to mycology).
In terms of genus Cannabis spp. there are three species: 1. Cannabis sativa L. f. ruderalis 2. Cannabis sativa L. subsp. indica 3. Cannabis sativa L. subsp. sativa
Ruderalis is not used in general. Breeding 'indica' to 'indica' or 'sativa' to 'sativa' gives one a cross, not a hybrid. That is a misnomer often found in the Cannabis spp. world. Within each species of Cannabis spp. there are many "land-race" strains, those which are pure and only inbreed. Those land-races are the original strains of Cannabis species. Just like I described for P.cubensis. Breeding strains of Cannabis 'sativa' to Cannabis 'indica' produces a intergenus hybrid....just like breeding P.azuras (sp?) and P.cubensis would provide an intergenus hybrid.
I hope that helps clear some things up.
Here are some examples of the use of "strain" and "species" in mycology. There are countless more examples online:
"Strain improvement in the cultivated mushroom Agaricus bisporus" -T. J. Elliott and F. A. Langton
Quote:
Summary Early attempts at genetic improvement in the cultivated mushroom Agaricus bisporus (Lange) Imbach were empirical, for little was understood of its natural breeding system. The mushroom is now known to be a secondarily homothallic species with a single multiallelic mating-type factor. This better understanding makes it possible to evaluate those breeding methods previously used and to suggest alternatives. Strain selection alone based on single spores, multispores or tissue culture may give improvement in the short term but it is unlikely to be as effective as methods involving controlled crossing. Mixing fertile strains may produce hybrids but it is difficult to identify them. It is better to use non-fertile isolates because only hybrids fruit. The earlier recognition of hybrids can be achieved using markers which are expressed in culture and genetic resistances may be especially useful in this respect. There is also a possible role for other Agaricus species which may be grown commercially and are more amenable to genetic manipulation than is A. bisporus.
"Taxonomic analysis of species and strains of oyster mushrooms Pleurotus (Fr.) Kumm. (Mycota, Basidiomycetes, Agaricales s. l.) collected at the Chair of Mycology and Algology, Moscow University. 1. Morphological peculiarities of Pleurotus species in vitro and in vivo." -Garibova, L. V., Zav'yalova, L. A., Insarova, I. D., Komarov, V. B., Lekomtseva, S. N., Chaika, M. N.
Quote:
Abstract: The morphological and cultural features of 6 species and 20 strains of Pleurotus mushrooms of different origin in the collection of the Chair of Mycology and Algology at Moscow University were studied. The species and strains differ in 4 parameters, namely structure of the aerial mycelium, zonality, colour, and diameter of colonies. Cluster analysis using the bootstrap showed that most strains of P. ostreatus formed a group with a high degree of similarity, including the Russian and foreign varieties of the oyster mushroom (Don 103, Don 112, Zommer and Florida) well known among the producer as well as strains of unknown origin. Individual groups were formed by 2 strains of P. ostreatus (from Germany and Saratov). The American strain US ME was found to be rather similar morphologically to P. pulmonarius and P. dryinus. The Far East strain DV 12 and unknown-origin strain KSH 52 proved to have characteristics similar to P. calyptratus. At the same time, the study identified some unrelated strains and species of the oyster mushroom: a strain from Baku, wild isolate of P. ostreatus, and the species P. eryngii. The main parameters playing an essential role in the fruitage of xylotrophic basidiomycetes (temperature, humidity and pH) were identified. The morphological features of basidia obtained in vitro in 6 species and 2 varieties of Pleurotus were studied in terms of 11 taxonomically significant morphological characters used to identify species in this subgroup. It was shown that, in vitro and in vivo, the shape of a cap, the colour of its cuticle, and the colour of the spore powder can be considered to be the most stable morphological characters of the Pleurotus species that have been studied. The results of the study demonstrate that the species diversity of xylotrophic basidiomycetes (micromycetes) can be preserved in a pure culture with subsequent reproduction of basidia, while preserving all the basic morphological characters of the species.
"Growing gourmet and medicinal mushrooms" -Paul Stamets
See Chapter 14: "Evaluating a Mushroom Strain"
"CULTURES: A Select Library of In Vitro Strains" -Paul Stamets
Quote:
The strains represented here are unique. Some are rare, not being available from culture libraries in North America, and potentially high yielding. We are constantly working with these strains, screening and maintaining cell lines. Fungi Perfecti is releasing the following strains for use on a conditional basis.
By purchasing these cultures, the customer agrees to the following terms. The purchaser is authorized by Fungi Perfecti to utilize our pure mushroom Spawn and Cultures for their own personal mycological experimentation, for the production of mushrooms for resale, and for the generation of additional mushroom spawn for the production of mushrooms for resale. The purchaser consents to use these strains for private use and not to generate spawn from them for resale or unauthorized distribution. Violators are subject to prosecution. The purchaser further agrees that mini-trials will be conducted prior to use in any commercial enterprise to assess their efficacy. The purchaser cannot and will not represent nor sell our strains in any form. Once shipped, Fungi Perfecti can not be responsible for the condition of any cultures. However, Fungi Perfecti will replace a culture if defective or should you lose it within one year of purchase at no additional charge.
Each strain is designated with the Stamets "P-value" scale, signifying the expansion of mycelium covering a 100 x 15mm petri dish (approximately 1,000–2,000 cell divisions). The first time a wild species is tissue cultured, the age is denoted as P-O. Thereafter each successive growth over the petri dish's surface is described in increments of P-l, etc. This library tries to maintain strains closest to their wild origins or closest to their peak fruiting potentials. Each culture comes in a 125 x 20mm glass test tube.
-------------------- Peace and Pasta
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potatonet
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Re: Methods for Fruiting Isolates [Re: Livingston]
#10470929 - 06/08/09 01:09 PM (14 years, 8 months ago) |
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Livingston you analyze shit a lot...
any you read a lot
when was the last time you took an IQ test
and will someone answer my in vitro question?
sorry to threadjack
-------------------- In the Quechua language of Peru there are over 1000 words for "potato"
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RogerRabbit
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Re: Methods for Fruiting Isolates [Re: Livingston]
#10471378 - 06/08/09 02:25 PM (14 years, 8 months ago) |
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Quote:
Livingston said: That is, P.cubensis from Amazon will be a different strain than one from Ecuador (for example). They will not have the same geno-type, or pheno-type, so they are different intraspecies strains...
Prove it. Mushroom spores travel on the wind everywhere, thus a cubensis from Brazil will be little or no different from Ecuador, Texas, Florida, etc. I've seen little repeated evidence of differences in so-called strains of mushrooms.
New growers do a multispore grow with one 'strain' and then purchase a different 'strain' for the next grow. They see two distinctly different presentations, thus they attribute it to strain. However, if they'd used the same syringe twice, they'd likely have seen the same difference in presentation. Such is the genetics of mushrooms grown from thousands of spores, each one with its own set of genetic information. Pot is a plant and is a whole different story, thus they don't compare. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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Livingston
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Re: Methods for Fruiting Isolates [Re: potatonet]
#10471998 - 06/08/09 04:19 PM (14 years, 8 months ago) |
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Quote:
potatonet said: Livingston you analyze shit a lot...
any you read a lot
when was the last time you took an IQ test
Thanks. Yea I do tend to read non-stop, it's kind of an addiction. I have a photographic memory of sorts so I can retain info well and I try to always read and empathize with both sides of the argument/discussion.
I haven't had my IQ tested since I was young. But when we are young our relative IQ is higher given our knowledge base. As we age our IQ tends to lower a bit. When I was tested I scored better than most but I'd rather talk about mushrooms! 
Quote:
and will someone answer my in vitro question?
RR would be better for that. But if you have two or more "fans" it doesn't sound like you have an isolate in the petri. Sure clone it! Why not? But I would try for around 10-20 true isolates and fruit them to find an elite. I have no idea if one that fruits in-vitro would be superior, my gut says not necessarily.
-------------------- Peace and Pasta
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Livingston
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Re: Methods for Fruiting Isolates [Re: RogerRabbit]
#10472146 - 06/08/09 04:44 PM (14 years, 8 months ago) |
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Quote:
RogerRabbit said:
Quote:
Livingston said: That is, P.cubensis from Amazon will be a different strain than one from Ecuador (for example). They will not have the same geno-type, or pheno-type, so they are different intraspecies strains...
Prove it. Mushroom spores travel on the wind everywhere, thus a cubensis from Brazil will be little or no different from Ecuador, Texas, Florida, etc. I've seen little repeated evidence of differences in so-called strains of mushrooms.
I thought I did with the references I gave? (especially the Russian one) Without very expensive (and illegal in the US) testing I can not prove it to you...but the same was said about microbes before the microscope was commonly available (poor analogy I know but you get the picture). There are volumes of data on the fact there are different strains of fungal species.
Cannabis spp. pollen travels *much* further than spores each time they flower. Spores travel around a few feet unless spread by the wind. Cannabis spp. pollen travels a couple dozen feet without wind.
With wind spores could travel say a mile or so due to their poor starting circumstances (next to ground in humid conditions with little wind). Spores are mainly spread by grazing animals who eat a blade of grass where for example some P.cubensis spores landed. Those spores will travel as far say 0-5 miles or a half a day before the animal craps them out (most grazing animals don't travel far each day).
Cannabis spp. pollen will travel miles with the wind, it’s been shown to travel tens of miles, as has other pollen. I know this because I am active in the non-GMO movement and GMO pollen pollution of heirloom (aka "land-race") strains is a major concern. Not to mention that male Cannabis spp. plants will release pollen constantly over a few weeks...
Quote:
New growers do a multispore grow with one 'strain' and then purchase a different 'strain' for the next grow. They see two distinctly different presentations, thus they attribute it to strain. However, if they'd used the same syringe twice, they'd likely have seen the same difference in presentation. Such is the genetics of mushrooms grown from thousands of spores, each one with its own set of genetic information.
IMO that's attributable to the naivety of most new growers. Breeders know you must work with a strain for a while to get to know its' characteristics and phenotype expressions, etc.
All black people look alike to white people, and visa versa. It's only the white person who is familiar with black people who sees obvious differences, and visa versa.
IMO getting to know a strain requires a lot of time and many mutli-spore and isolate grows...
Quote:
Pot is a plant and is a whole different story, thus they don't compare. RR
I disagree. Traditionally "land-race" Cannabis spp. species strains, ex: 'C.indica Afghan' (similarly: 'P.cubensis Ecuador') are grown from so-called "open pollination". That is where hundreds or thousands of male plants are allowed to spread pollen to hundreds or thousands or more female plants via. wind. This is how one stabilizes a Cannabis spp. species strain...and how one stabilizes a P.cubensis strain (via inbreeding from a interspecies cross for example). Those two examples are simply where people are trying to replicate Natures' breeding methods on a much less grand scale.
-------------------- Peace and Pasta
Edited by Livingston (06/08/09 04:56 PM)
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Lucid_Euphoria
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Re: Methods for Fruiting Isolates [Re: Livingston] 1
#10472731 - 06/08/09 06:39 PM (14 years, 8 months ago) |
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I'm no biologist nor am I a mycologist. However, I see RR's point in that plants and fungi mate and pass on thier genetic code in completely different ways.
I'm pretty sure spores from a fruit contain thousands of different genetic code, as well as being different "sex types", such as male and female in the human species, except fungus have hundreds and possibly thousands of "sex types". After spore germination, the monokaryotic mycelium must "mate" with a compatible monokaryotic mycelium sex type, this fusing of seperate mycelium colonies is known as Dikartotization, producing a dikaryotic mycelium.
The genetic code of the spores isn't programmed until the formation of the basidium I believe. I could be wrong there, but I think thats how I read it works.
So basically, within the P. Cubensis strain there are thousands of types of genetic code which can all produce differing results. Which is what I think RR was trying to point out. That those newbie growers using MS inoculation could get thousands or characteristics from one grow to another due to the way the fungus mates. So Equador, Texas, Cambodian, whatever you want to call it, all have the same huge variety of genetic possibilities. So the medium on which it is grown, isn't a huge factor in the characteristics of the fruits it will produce, it is primarily genetic. Of course ideal conditions and nutrients will insure a higher bioefficiency, but the characteristics are purely genetic.
-------------------- PAN CYAN & AZURE FOR TRADE "If ignorance is bliss, THEN KNOCK THE SMILE OFF MY FACE!" - Zach de la Rocha "Today a young man on acid realized that all matter is merely energy condensed to a slow vibration, that we are all one consciousness experiencing itself subjectively, there is no such thing as death, life is only a dream, and we are the imagination of ourselves. Here's Tom with the weather!" -Bill Hicks-
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