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http://indianaquariumhobbyist.com/community/ :: View topic - Aquatic Plants Tissue Culture
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Aquatic Plants Tissue Culture
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karthik_nambiar
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PostPosted: Wed Jul 25, 2012 3:41 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 wow,  this  is  getting  intresting  (although  did  not  like  biology  &  Science  from  school).  
 
 Chemistry  used  to  be  CHEE  masti.  
 
 Now  i  undertand  why  people  say  "HISTORY  Repeats"  !!!
 
 Would  love  to  help  in  what  ever  way  possible  &  LEARN  from  this  PRACTICAL  experience...
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Ravabi
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PostPosted: Wed Jul 25, 2012 3:45 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Hi  Madan
 
 I  am  very  much  interested  in  it.   
 
 As  a  UG  student  I  was  able  to  culture  Duranta  gold  (A  common  Fluorescent  green  hedge  plant).  I  also  tried  out  with  Ficus  guavajava  (Guava  tree),  but  was  not  successful  due  to  the  high  phenol  content.  
 
 How  different  is  it  for  aquatic  plants?  Could  you  forward  me  some  literature  on  it.
 
 Starting  out
 1.  I  have  some  100  ml  sample  containers  (Tar  sons)  which  I  use  for  storing  water  samples.  They  may  be  resistant  possibly  till  70  or  80  and  are  pretty  cheap.  Don't  know  it  they  will  withstand  Autoclave  treatments.  
 http://www.tarsons.in/?a=c&s=510010.  Will  also  bring  them  during  the  next  ASK  meet.
 2.  Dimethyl  sulfoxide  and  Agarose  can  be  sourced  out  from  chemical  stores  like  Vasa  Scientific  (Malleshwaram  /  S.P.  Road)
 3.  Scalpel,  Forcep,  Surgical  gloves  can  also  be  sourced  from  Vasa
 4.  Multi-Vitamin  tablets  and  antibiotics  must  be  the  more  expensive  part.  We  can  order  them  based  on  the  needs.
 
 ravi
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sandeepraghuvanshi
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PostPosted: Wed Jul 25, 2012 3:48 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Which  plants  are  you  going  to  start  with.
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Madan
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PostPosted: Wed Jul 25, 2012 4:00 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Sandeep  -  Crypts,  one  or  two  types  actually  !  Might  try  Narrow  leaf  Java  Fern  also,  but  that  needs  a  liquid  medium.
 
 You  need  emersed  specimens  for  tissue  culture,  submerged  specimens  will  simply  not  do.
 Too  bad  I  replanted  all  my  emersed  Crypts  just  a  few  days  ago,  didn't  know  I'd  end  up  with  a  few  days  spare  time.
 
 Ravi  -  There's  hardly  any  literature  worth  the  while  about  aquatic  plants.  Thanks  for  the  tip  about  agarose  and  Dimethyl  sulfoxide.
 What  was  the  biocide  you  added  to  the  media  ?  Mercuric  chloride  is  one  but  that's  toxic,  I'd  rather  take  my  chances  without  any  biocide.
 
 I  feel  we  have  to   go  with  glass  bottles  unless  someone  find  us  a  good  source  for  microwave  safe  plastic  bottles  and  lids.
 Any  source  for  glass  bottles  ?
 
 If  anyone  has  emersed  grown  plants  that  could  also  be  tried.
 
 Which  is  better  Isopropyl  alcohol  or  Ethanol  for  sterilising  ?
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rasikanayak
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PostPosted: Wed Jul 25, 2012 4:02 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Waaaaaah,  I  am  going  to  miss  this   Sad   Sad  
 
 Guys  from  Bangalore  North,  Try  Vasa  on  11th  cross,  Malleshwaram.  Glass  bottles  and  jars  too  available  there.
 
 Regards,
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Cherry
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PostPosted: Wed Jul 25, 2012 4:03 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Madan  Ji...normally  for  culturing,cotton  rolled  tight  ,  covered  with  aluminium  foil  is  used  to  cap  the  cultures.
 Can  this  be  used?  Metal  caps  will  leave  the  bottles  airtight  not  letting  the  cultures  to  breathe.
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Madan
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PostPosted: Wed Jul 25, 2012 4:07 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Shanai,  aluminium  foil  will  do  the  same  thing  as  a  metal  cap.  Bottles  need  to  be  closed  tight  to  avoid  contaminants.
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bettaji
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PostPosted: Wed Jul 25, 2012 4:08 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Will  this  work  on  ferns  too  ?
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menonabhishek
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PostPosted: Wed Jul 25, 2012 4:11 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 This  will  be  interesting  ...
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Madan
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PostPosted: Wed Jul 25, 2012 4:13 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Shanai,  thanks  a  lot  !
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Cherry
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PostPosted: Wed Jul 25, 2012 4:19 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Madan  Ji,ethanol  is  preferably  better  as  I  learnt  in  my  little  experience  in  culturing.
 Also,  rightly  said  by  you,  mercuric  chloride  will  be  toxic,  better  to  start  without  biocide,  instead  Sodium  or  calcium  hypochlorite  can  be  tried,  but  we  need  to  check  more  on  them  before  using.
 
 Auxins  can  be  added  for  the  explant  growth  and  regulation,  I  believe  it  should  be  easily  available.
 
 But  the  most  difficult  step  would  be  storing  the  cultures  as  they  will  need  a  complete  aseptic  place  to  grow  atleast  for  a  month,or  the  enriched  media  will  attract  a  lot  of  foreign  bodies  Sad


Last edited by Cherry on Wed Jul 25, 2012 4:26 pm; edited 1 time in total
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Cherry
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PostPosted: Wed Jul 25, 2012 4:21 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

                                                   
Madan  wrote  (View  Post):                
Shanai,  thanks  a  lot  !                

 
 Most  welcome   Madan  Ji:)
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Madan
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PostPosted: Wed Jul 25, 2012 4:41 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Shanai,  I  have  auxins  dissolved  in  ethanol  ready  for  use.
 I  need  Dimethyl  sulfoxide  to  dissolve  Cytokinins  and  make  a  stock  solution.
 
 I  don't  have  Giberellic  acid,  but  that's  not  needed  for  now.
 I  should  anyway  be  getting  Giberellic  acid  in  a  few  days.
 
 It's  best  not  to  use  commercial  plant  hormones  for  aquariums  like  the  one  sold  by  Azoo,  as  we  don't  know  how  many  fingers  have  contaminated  the  stuff.
 
 You  are  right  I  don't  expect  success  the  first  time,  but  if  enough  of  us  start  off,  we  should  be  able  to  get  there  pretty  soon.
 
 It  is  better  people  start  out  culturing  a  few  plants  emersed,  which  is  very  easy,  so  that  they  can  readily  experiment.
 
 This  thread  should  serve  as  a  good  place  to  put  in  all  the  info  we  gather  as  we  go  along.
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Cherry
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PostPosted: Wed Jul 25, 2012 4:56 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Definitely  Madan  Ji,  your  start  will  lead  us  to  a  success  Thumb Up
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Ravabi
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PostPosted: Wed Jul 25, 2012 5:03 pm Post subject: Re: Aquatic Plants Tissue Culture Reply with quote

 Madan,  As  Nayak  pointed  out,  Vasa  Scientific  in  Malleshwaram  is  very  much  accessible  to  me.  Will  check  for  the  prices  of  both  Agarose  and  Dimethylsulfoxide.
 We  used  Kanamycin,  which  is  detrimental  to  most  bacteria  (main  contaminants).  I  will  also  check  for  other  alternatives  from  my  friends.  
 We  added  commercial  macro  and  micro  nutrients  (Not  Azoo)  to  agar  (similar  to  dosing  of  ferts),  alongside  with  Auxin  and  Cytokinin.  
   
 Shanai,  As  you  suggested,  the  biggest  problem  will  be  the  storage  and  aseptic  condition.  We  can  always  fume  dedicated  boxes  (few  small  ones)  with  formalin  and  store  it.  Wont  it  work?
 
 The  most  effective  agent  for   cleaning  the  explants  would  be  to  rinse  them  with  Methanol/  dilute  Hg2cl2  for  a  minute  or  so.
 
 Ravi
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