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socratesmind
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Registered: 02/22/02
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Agar isolations ? 1
#872206 - 09/09/02 09:53 AM (21 years, 6 months ago) |
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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...
-------------------- 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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Anno
Experimenter
Registered: 06/17/99
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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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socratesmind
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Re: Agar isolations ? [Re: Anno]
#872848 - 09/09/02 03:32 PM (21 years, 6 months ago) |
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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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DinoMyc
Ipsa scientiapotestas est
Registered: 11/13/99
Posts: 1,080
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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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Anno
Experimenter
Registered: 06/17/99
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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 06:16 PM)
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Zen Peddler
Registered: 06/18/01
Posts: 6,379
Loc: orbit
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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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Peace_Patrol
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Re: Agar isolations ? [Re: Zen Peddler]
#873486 - 09/09/02 11:40 PM (21 years, 6 months ago) |
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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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socratesmind
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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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Peace_Patrol
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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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socratesmind
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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
-------------------- 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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DinoMyc
Ipsa scientiapotestas est
Registered: 11/13/99
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Last seen: 17 years, 9 months
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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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Champion des Champignons
long standing member;)
Registered: 07/26/00
Posts: 2,680
Loc: Alba
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Re: Agar isolations ? [Re: Zen Peddler]
#875410 - 09/10/02 05:01 PM (21 years, 6 months ago) |
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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?
-------------------- --------------------------------------------------- hmmm........
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bluepoo
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Registered: 03/27/02
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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
-------------------- ----------------------------------------------------------- the above is what the litel leprecans told me to say and should be regarded as such -----------------------------------------------------------
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Zen Peddler
Registered: 06/18/01
Posts: 6,379
Loc: orbit
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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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Visigoth
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Re: Agar isolations ? [Re: Zen Peddler]
#880313 - 09/12/02 10:06 AM (21 years, 6 months ago) |
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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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socratesmind
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Re: Agar isolations ? [Re: Visigoth]
#885243 - 09/16/02 07:34 AM (21 years, 6 months ago) |
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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
-------------------- 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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Anno
Experimenter
Registered: 06/17/99
Posts: 24,166
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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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TripsAreForKids
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Re: Agar isolations ? [Re: Zen Peddler]
#885983 - 09/16/02 02:59 PM (21 years, 6 months ago) |
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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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socratesmind
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Re: Agar isolations ? [Re: Anno]
#886489 - 09/16/02 05:52 PM (21 years, 6 months ago) |
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Thanks a mucho Anno i think i'm going to B&N to get my book special ordered tomm
-------------------- 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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Anno
Experimenter
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Posts: 24,166
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