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Offlinesocratesmind
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Agar isolations ?
    #872206 - 09/09/02 11:53 AM (15 years, 21 days ago)

Whenever i receive a syringe in the mail from any of these vendors it is a particular species. But not so much a pure strain of that species? And this can only be obtained by isolating and reisolating in agar mediums until its really rhizemorphic growth? Then that is when you have the true isolation of a pure species??? Can you reisolate this pure/semi pure isolation to maybe make it tend to grow faster (i.e. take the first ropey looking strand in the mycellium and isolate over and over) Would this train the mycellium to colonize faster or is there a ceiling limit on how fast that the mycellia can colonize? Really interested in agar cultures and isolations of pure strains...


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Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.


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OfflineAnnoA
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Re: Agar isolations ? [Re: socratesmind]
    #872406 - 09/09/02 01:31 PM (15 years, 21 days ago)

I strongly suggest you to buy the Mushroom Cultivator( Link at www.shroomery.org/books ). It will answer all and more of your questions.


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Offlinesocratesmind
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Re: Agar isolations ? [Re: Anno]
    #872848 - 09/09/02 05:32 PM (15 years, 20 days ago)

Could you maybe post the TOC or table of contents for me to see what all the book contains. I'm really interested in fungi in all aspects not jsut the magic producing ones...


--------------------
Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.


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OfflineDinoMyc
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Re: Agar isolations ? [Re: socratesmind]
    #872918 - 09/09/02 06:02 PM (15 years, 20 days ago)

Trust us, just buy it.
That and GGMM together make up a fairly complete collection of what you need to know..


--------------------
If I made affront, I apologize.
If I made affirmation, I apologize.
I merely came to listen, came to say.


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OfflineAnnoA
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Re: Agar isolations ? [Re: socratesmind]
    #873135 - 09/09/02 08:15 PM (15 years, 20 days ago)

Did you check the link I gave you at all??

One more time:

www.shroomery.org/books

At the bottom of the page......

There you will find the table of contents......


Edited by Anno (09/09/02 08:16 PM)


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InvisibleZen Peddler
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Re: Agar isolations ? [Re: socratesmind]
    #873459 - 09/10/02 01:00 AM (15 years, 20 days ago)

If you continue to reisolate the strain systematically on the same media it will get pretty damn fast, but it will also become selective and adapt the media - making its growth on other nutrients other than MEA, etc agar less viable.


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OfflinePeace_Patrol
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Re: Agar isolations ? [Re: Zen Peddler]
    #873486 - 09/10/02 01:40 AM (15 years, 20 days ago)

So, would it be a good idea to isolate some rhizos, then grow it out on wood, and then take a chunk of colonised wood and put this on some agar again? Or would this have no effect on helping reduce substrate preferences?


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Offlinesocratesmind
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Re: Agar isolations ? [Re: Peace_Patrol]
    #873506 - 09/10/02 02:19 AM (15 years, 20 days ago)

What i think bluemeanie means is that the more and more faster u stimulate it's growth its just gonna grow that much more picky when selecing a substrate other than MEA. So it's gonna be a good colonizer yes in agar and might not colonize lets say BRF or grains or stuff like that. Future experiment would be nice so we can have hard data.


--------------------
Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.


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OfflinePeace_Patrol
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Re: Agar isolations ? [Re: socratesmind]
    #873513 - 09/10/02 02:38 AM (15 years, 20 days ago)

Yeah, I understood that, so I was asking, would it be a good idea to avoid this single-substrate preference by sometimes growing it out on wood or some other substrate before switching back to agar.


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Offlinesocratesmind
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Re: Agar isolations ? [Re: Peace_Patrol]
    #874603 - 09/10/02 01:04 PM (15 years, 20 days ago)

Well one way i was thinking that you could accomplish this to make it diverse on substrates is gradually introduce your target substrates at certain levels in the isolations and increase the substrate level with each interval.  So it will be at least use to seeing/eating that particular substrate.  This should be a nice experiemnt to try.  I'd love to do but i need to get my glovebox runnin and get some agar/petri dishes and a few other goodies :frown:


--------------------
Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.


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OfflineDinoMyc
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Re: Agar isolations ? [Re: socratesmind]
    #875069 - 09/10/02 04:28 PM (15 years, 19 days ago)

Glovebox is old school.. way old school.. if your spending money, go with a good flow hood.
have you read tmc and ggmm yet?
do so.. it will answer all these questions..


--------------------
If I made affront, I apologize.
If I made affirmation, I apologize.
I merely came to listen, came to say.


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InvisibleChampion des Champignons
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Re: Agar isolations ? [Re: Zen Peddler]
    #875410 - 09/10/02 07:01 PM (15 years, 19 days ago)

surely the problem with becoming dependent, or at least preferring a particular substrate only occurs through successive multispore innoculations?
ie if you make a multispore innoc on eg MEA, fruit the fastest growing substrain, then use the spores from this substrain in a new multispore on the same medium, fruit the fastest of these etc, then eventually, theoretically at least, you could end up with a sub-sub.....substrain that will only grow on MEA and nothing else?
If a particular substrain has the required DNA to produce the enzymes to digest a wide variety of substrates then surely no end of cloning is going to change this? Obviously if a substrain "gets used to" a particular medium it will take a while for it to start producing the relevant enzymes which were previously unneeded, eg how it takes a few days for mycelium to adapt to peroxidated agar, but I don't see how it could lose this adaptability altogether without genetic change being involved.
or am I missing something?


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---------------------------------------------------
hmmm........


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Offlinebluepoo
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Re: Agar isolations ? [Re: socratesmind]
    #875558 - 09/10/02 08:35 PM (15 years, 19 days ago)

the thing is there is a limited amount of time that a strain will work it doesent die but it becomes useless this is why pepole keep stock of strains in a fridge once you get it working well do this and that book has a lot of info on it


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the above is what the litel leprecans told me to say and should be regarded as such
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InvisibleZen Peddler
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Re: Agar isolations ? [Re: Champion des Champignons]
    #875904 - 09/11/02 01:44 AM (15 years, 19 days ago)

Stamets suggests that each transfer of an isolate should be done on new nutrient bases for agar plates otherwise the mycelia will quickly adapt to become selective. Im quoting stamets when im talking here, not from personal experience as ive noticed little difference between second generation and 12th generation on MEA


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OfflineVisigoth
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Re: Agar isolations ? [Re: Zen Peddler]
    #880313 - 09/12/02 12:06 PM (15 years, 18 days ago)

I have a question, when the agar plate has completely colonized, and it's time to examine the mycelium carefully in order to find a rhizomorphic section to transfer to the next plate. Can u just take the top off to examine w. out having to worry about contams, meaning, once it's colonized, is it pretty resistant to contams just like a colonized jar?? My friend has a plate that will be done soon and he wanted to know about this. Thanks!!!!

Vis


--------------------
"15 minutes in freezing conditions?!?! You'd become a popsicle before you made that!!!"


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Offlinesocratesmind
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Re: Agar isolations ? [Re: Visigoth]
    #885243 - 09/16/02 09:34 AM (15 years, 14 days ago)

Better safe than sorry why try to contam your culture at this point just take the same precautions and be careful and remember to store your pure culture in a slant so u can always go back to it :smile: 


--------------------
Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.


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OfflineAnnoA
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Re: Agar isolations ? [Re: socratesmind]
    #885285 - 09/16/02 09:56 AM (15 years, 14 days ago)

code:

Mushroom Cultivator
TABLE OF CONTENTS


FOREWORD by Dr. Andrew Weil xi
PREFACE xii
I.INTRODUCTION TO MUSHROOM CULTURE 1
An Overview of Techniques for Mushroom Cultivation 3
Mushrooms and Mushroom Culture 4
The Mushroom Life Cycle 6
II.STERILE TECHNIQUE AND AGAR CULTURE 15
Design and Construction of a Sterile Laboratory 16
Preparation of Agar Media 19
Starting A Culture from Spores 23
Taking a Spore Print 23
Techniques for Spore Germination 24
Characteristics of the Mushroom Mycelium 25
Ramifications of Multispore Culture 25
Sectoring: Strain Selection and Development 31
Stock Cultures: Methods For Preserving Mushroom Strains 37
III.GRAIN CULTURE 41
The Development of Grain Spawn 42
Preparation of Grain Spawn 45
Spawn Formulas 46
Inoculation of Sterilized Grain from Agar Media 48
Inoculation of Sterilized Grain from Grain Masters 49
Alternative Spawn Media 54
Liquid Inoculation Techniques 55
Incubation of Spawn 57
IV.THE MUSHROOM GROWING ROOM 61
Structure and Growing Systems 62
Structure 63
Shelves 64
Trays 65
Environmental Control Systems 66
Fresh Air 66
Fans 68
Air Ducting 70
Filters 70
Exhaust Vents 72
Heating 73
Cooling 73
Humidification 74
Thermostats and Humidistats 74
Lighting 74
Environmental Monitoring Equipment 76
V.COMPOST PREPARATION 77
Phase I Composting 78
Basic Raw Materials 78
Supplements 79
Formulas 81
Ammonia 82
Carbon:Nitrogen Ratio 83
WaterAir 83
Pre-Wetting 84
Building the Pile 85
Turning 87
Temperature 88
Long Composting 89
Short Composting 90
Synthetic Compost Procedure 91
Composting Tools 92
Characteristics of the Compost at Filling 93
Supplementation at Filling 95
Phase II Composting 96
Basic Air Requirements 97
Phase II Room Design 98
Filling Procedures 98
Depth of Fill 99
Phase II Procedures: Trays or Shelves 100
Phase II in Bulk 101
Bulk Room Design Features 102
Bulk Room Filling Procedures 104
Bulk Room Phase II Program 104
Testing for Ammonia 104
Aspect of the Finished Compost 105
Alternative Composts and Composting Procedures 106
Sugar Cane Bagasse Compost 106
The Five Day Express Composting Method 106
VI.NON-COMPOSTED SUBSTRATES 109
Natural Culture 110
Wood Based Substrates 114
Straw 117
VII.SPAWNING AND SPAWN RUNNING IN BULK SUBSTRATES 121
Moisture Content 122
Substrate Temperature 122
Dry Weight of Substrate 122
Duration of Spawn Run 124
Spawning Methods 124
Environmental Conditions 125
Super Spawning 126
Supplementation at Spawning 126
Supplementation at Casing 126
VIII.THE CASING LAYER 127
Function 128
Properties 129
Materials 130
Formulas and Preparation 132
Application 133
Casing Colonization 135
Casing Moisture and Mycelial Appearance 137
IX.STRATEGIES FOR MUSHROOM FORMATION (PINHEAD INITIATION) 139
Basic Pinning Strategy 140
Primordia Formation Procedures 141
The Relationship Between Primordia Formation and Yield 146
The Influence of Light on Pinhead Initiation 147
X.ENVIRONMENTAL FACTORS: SUSTAINING THE MUSHROOM CROP 149
Temperature 150
Flushing Pattern 150
Air Movement 152
Watering 154
Harvesting 155
Preserving Mushrooms 156
XI.GROWING PARAMETERS FOR VARIOUS MUSHROOM SPECIES 159
Agaricusbitorquis 161
Agaricus brunnescens 164
Coprinus comatus 168
Flammulina velutipes 172
Lentinus edodes 176
Lepista nuda 180
Panaeolus cyanescens 183
Panaeolussubbalteatus . 186
Pleurotus ostrealus (Type Variety) 189
Pleurotus ostreatus (Florida Variety) 193
Psilocybe cubensis 196
Psilocybe cyanescens 200
Psilocybe mexicana 204
Psilocybe tampanensis 207
Stropharia rugoso-annulata 210
Volvariella volvacea 214
XII.CULTIVATION PROBLEMS AND THEIR SOLUTIONS:
TROUBLE SHOOTING GUIDE 217
Sterile Technique 219
Agar Culture 219
Grain Culture 220
Compost Preparation 223
Phase I 223
Phase II 224
Spawn Running 226
Case Running 227
Mushroom Formation and Development 229
Pinhead Initiation 229
Cropping 231
XIII.THE CONTAMINANTS OF MUSHROOM CULTURE:
IDENTIFICATION AND CONTROL 233
A Key to the Common Contaminants of Mushroom Culture 238
Virus (Die-Back Disease) 244
Actinomyces (Firefang) 246
Bacillus (Wet Spot) 248
Pseudomonas (Bacterial Blotch & Pit) 252
Streptomyces {Firefang) 255
Alternaria (Black Mold) 257
Aspergillus (Green Mold) 259
Botrytis (Brown Mold) 262
Chaetomium (Olive Green Mold) 264
Chrysosporium (Yellow Mold) 266
Cladosporium (Dark Green Mold) 268
Coprinus (Inky Cap) 270
Cryptococcus (Cream Colored Yeast) 273
Dactylium (Cobweb Mold) 275
Doratormyces (Black Whisker Mold) 277
Epicoccum (Yellow Mold). 279
Fusarium (Pink Mold) 281
Geotrichum (Lipstick Mold) 284
Humicola (Gray Mold) 286
Monilia (White Flour Mold) 288
Mucor (Black Pin Mold) 290
Mycelia Sterilia (White Mold) 292
Mycogone (Wet Bubble) 294
INeurospora (Pink Mold 296
Papulospora (Brown Plaster Mold) 298
Penicillium (Bluish Green Mold) 300
Rhizopus (Black Pin Mold) 302
Scopulariopsis (White Plaster Mold) 304
Sepedonium (White or Yellow Mold) 306
Torula (Black Yeast) 308
Trichoderma (Forest Green Mold) 310
Trichothecium (Pink Mold) 313
Verticillium (Dry Bubble) 315
XIV.THE PESTS OF MUSHROOM CULTURE 319
Mushroom Flies 320
Fly Control Measures 320
Sciarid Fly 321
Phorid Fly 323
Cecid Fly 325
Mites 328
INematodes (Eelworrns) 331
XV.MUSHROOM GENETICS 333
Reproductive Strategies 336
Implications for Culture Work 338
APPENDICES 343
I.Medicinal Properties of Mushrooms 345
II.Laminar Flow Systems 347
III.The Effect of Bacteria and Other
Microorganisms on Fruiting 253
IV.The Use of Mushroom Extracts to Induce Fruiting 357
V.Data Collection and Environmental Monitoring Records 359
VI.Analyses of Basic Materials Used in Substrate Preparation 369
VII.Resources For Mushroom Growing Equipment and Supplies 384
VIII.English to Metric Conversion Tables 386
GLOSSARY 389
BIBLIOGRAPHY 397
INDEX 409
PHOTOGRAPHY AND ILLUSTRATION CREDITS 414
ACKNOWLEDGEMENTS. 415



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InvisibleTripsAreForKids
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Re: Agar isolations ? [Re: Zen Peddler]
    #885983 - 09/16/02 04:59 PM (15 years, 13 days ago)

In reply to:

If you continue to reisolate the strain systematically on the same media it will get pretty damn fast, but it will also become selective and adapt the media - making its growth on other nutrients other than MEA, etc agar less viable.




Well, could you make some brown rice flour agar and keep isolating on it, then spawn it to some whole brown rice jars? Won't it be totaly suited for that substrate? Thus growing faster and better on it.


--------------------
Everything posted above was out of fun, none should be taken serious. I am currently under the influence so take that into consideration.


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Offlinesocratesmind
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Re: Agar isolations ? [Re: Anno]
    #886489 - 09/16/02 07:52 PM (15 years, 13 days ago)

Thanks a mucho Anno i think i'm going to B&N to get my book special ordered tomm :smile:


--------------------
Prohibition will work great injury to the cause of temperance. It is a species of intemperance within itself, for it goes beyond the bounds of reason in that it attempts to control a man's appetite by legislation, and makes a crime out of things that are not crimes. A Prohibition law strikes a blow at the very principles upon which our government was founded.
- Abraham Lincoln: Speech in the Illinois House of Representatives, Dec 18, 1840.


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OfflineAnnoA
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Re: Agar isolations ? [Re: socratesmind]
    #886903 - 09/17/02 12:21 AM (15 years, 13 days ago)

>i think i'm going to B&N

Is the book cheaper there?
I looked and the book is not available there
http://search.barnesandnoble.com/booksearch/isbnInquiry.asp?userid=0DDKSXPISD&isbn=0961079800

You can order it at Amazon:

http://www.amazon.com/exec/obidos/ASIN/0961079800/minmed-20/ 


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