As agar as a gelling agent in plant tissue culture is expensive, I would like find an alternative for agar.
OK, Gum katira, Sago or Cassava starch can be considered as alternative low-cost gelling agents, but preparing of these products are not easy for everyone. If we can find a product that be find in everywhere, it will be better.
And if we use cassava starch as alternative to Agar, we may need to change sugar concentration in the media!
Try 'isabgol' which gels well at 3.5%. Isabgol is the husk of the seeds of Plantago ovata. It is readily available as a popular laxative in India, but should be available elsewhere too.
You can also have a look at my paper on low cost gelling agents:
Vinod S. Gour and Tarun Kant (2011). Efficacy of Low Cost Gelling Agents and Carbon Source Alternatives during In Vitro Rooting of Balanites aegyptiaca and Phyllanthus emblica Microshoots. Tree and Forestry Science and Biotechnology, 5 (Special Issue 1): 58-60
Hi, Danial. As suggested by Tarun Kant, I would also suggest you to try 'Isabgol (Psyllium Seed Husk)'. You can try the methodology in my paper as well. I have attached the publication. All the Best! If you have any queries you are always welcome!
Article Low-cost media for in vitro conservation of turmeric (Curcum...
You can use alginate (10 US/k. But, to form the gel you need to disolve the alginate (1%) and add it to a calcium cloride solution (10%). The gel is formed instantly. You can olso use dialisis bags, and introduce it to a calcium cloride solution (10%) over night. Then you cut the gel to the sice you want and your plant tissue will grow perfectly.
hi dear Dr. Kahrizi. Recently, researchers try to find a suitable substrate instead of agar, because of high price of pure grade agar and there are some doubts about its nontoxic nature. Moreover, the exclusive use of agar may result in over exploitation of its resources (Jain and Babbar, 2005; Deb and Pongener, 2010). Different materials such as, starch from various sources (Sorvari and Schieder, 1987;
Zimmerman, 1995; Dabai and Muhammad, 2005), starch with low concentration of gelrite (Zimmerman, 1995), combination of starch: semolina: potato powder (2:1:1) (Prakash et al., 2002), Isubgo (Babbar and Jain, 1998;
Jain et al., 1997), sago (Bhattacharya et al., 1994), Xanthan gum (Babbar and Jain, 2006) and guar gum (Babbar et al., 2005; Jain et al., 2005) were used as alternative gelling agents. please find the attached file.
hi dear Dr. Kahrizi. Recently, researchers try to find a suitable substrate instead of agar, because of high price of pure grade agar and there are some doubts about its nontoxic nature. Moreover, the exclusive use of agar may result in over exploitation of its resources (Jain and Babbar, 2005; Deb and Pongener, 2010). Different materials such as, starch from various sources (Sorvari and Schieder, 1987;
Zimmerman, 1995; Dabai and Muhammad, 2005), starch with low concentration of gelrite (Zimmerman, 1995), combination of starch: semolina: potato powder (2:1:1) (Prakash et al., 2002), Isubgo (Babbar and Jain, 1998;
Jain et al., 1997), sago (Bhattacharya et al., 1994), Xanthan gum (Babbar and Jain, 2006) and guar gum (Babbar et al., 2005; Jain et al., 2005) were used as alternative gelling agents. please find the attached file.
I use Food Grade Agar for my tissue culture works , I bought 500g for 400,000 Rial (800,000 Rial per Kilo) Not very cheap but cheaper than Agar from Merk or other companies .
I use 5-6g/L Agar for 16*100 test tubes and 50ml falcon tubes and it give a good gel for bigger culture vessels 7g is better , it give a good clear gel as clear as the gel of Merk's Agar ! you can get it from confectionery supplies shop .
The concentration of agar in medium depend on the experiment but generally I used 7-8 g/L phytoagar with the tissue culture experiment and it is giving hard gel. the best amount I used 1kg phtoagar from sigma company/UK.
Dear Dr. Kahrizi. I think what kind of agar to be used depends on what you culture, for some plants culture, there sometimes is specific. The better choose may be that you should try the agar from high price to low price. Unfortunately, what I used for picea asperate is gelling gum which is a very expensive one but suitable for culture.
Good Luck !
Look for Colagar. It is semi porecessed seawweds. So, you have to extract the agar.
You boil the algae, filter and it is the fitrated solution that you can use as culture medium.
We use Agargel(Sigma a mix of pytogel and agar) at 0.7% and this gives a good strong gel which goes far due to its low percentage concentration and saves a bit without compromising quality.
Agar verses Phytagel:
Phytagel (Sigma P8169 @ $256.00/KG) use at 1.5 to 2.5 g/L; much clearer/more transparent than agar; requires calcium or other divalent cation to gel (not a problem with plant culture media but may be a concern with some bacteriologic media); increasing calcium concentration increases gel strength.
Agar (plant tissue culture grade; Sigma A7921 @ $442.00/KG) use at 6-12 g/L
Phytagel costs $0.38 to $0.64 / liter of media
Plant Culture-grade Agar costs $2.65 to $5.30 / liter of media
In many plant industry companies, we have started not to use agar anymore. For example, using rock-wool as a medium, instead of agar, is gaining wide acceptance for many species. For your consideration - see Fig 9.5 and Fig 9.9 (taken from Hew and Yong, 2004)
You can use sterilized stone pebbles or brick chips, or folded blotting paper...it is a low cost method..
What we have been calling Gelrite is now Gelzan. The surviving Gelrite is a hand sanitizer.
Phytagel is better and cheaper option instead of Agar, for maize tissue culture .
When we talk about a cheaper alternative of agar, we cannot compare simply the cost of agar and alternative material. We have to consider the concentration used in the medium without affecting protocol efficiency. If a cheaper alternative reduces the efficiency of protocol and ultimately cost of plant production (Cost per plant) is not decreases cannot be considered as cheaper alternative. Thus a cheaper alternative may be effective in one protocol but the same material may be costlier in other tissue culture protocol.
I recently reviewed a paper. The author had used and recommended isubgol obtained from the seeds of Plantago.
It seemed to be very cheap.
Has anyone tried polyacrylamide gel? all diff grades from snow to chunks to round balls. The ag grade is K-salt and cross-linked more to persist for months in the field use. vs. Na-acrylamide.
Polyacrylamide gel is better fir keeping water away from baby's butt. I tried it for sprouting seeds without success/
china makes a ton of cheap, crappy stuff. but there are some US players. there could be phytotoxicity from the Na or (god forbid) free acrylamide. used in forestry, agronomic crops, and in nursery--like moisture control potting mix. probably needs fine tuning for use in micropropagation or greenhouse transitioning. sometimes called Hydrophilic Hydrogels. Some made from starch instead of acrylamide, some biodegradable.
Rodney
A poly-starch hydrogel has possibilities. Where does one find it?
Free article on Springerlink
http://link.springer.com/article/10.1023%2FA%3A1010267929629#page-1
We cannot say alternative to agar but alternative support material to use in place of agar, it is cotton, you can try it in quantity which can support explant or cultured tissue to be elevated enough to grow and absorb nutrients through cotton. it works well, it has been used in tissue culture of sugarcane and published as well.
Larry: starch gels hard to find. google Reon/Zeba, BioSAP™ from ADM. PAM from diapers going into the landfill may be creating a hazard. starch gel guys are trying to replace the PAM market in diapers. Diaper use still no. 1. Ag/nursery use is less.
Type of gelling agent depends on the mode of experiment. In almost all cases agar is used as gelling agent. But if want to study rooting process, it is better to use gelrite and phytagel.
Guar gum--about 1:1 with agar. It's supposed to be more fluid with superior diffusion, leading to better plant response. The bonus is the cost cutting, although the cost has gone up 20 fold since oil drillers started using it. And it is generally impure, may need cleanup prior to use.
QWe use wheat starch at 50 g/l and it was a success for many species.
I used 50g/l cassava starch in combination with alittle agar, I did this for my msc om banana tissue culture and has also worked on sweet potato and eucalyptus
For grapevine, I used perlite with liquid MS media (not above the level of perlite, otherwise the plant will be moving).
It works very well for cuttings, to make them root and get plantlets.
Please tell us what you do exactly, cause if you make callus cultures, this won't work of course.
I would say, several types of gelling agents should be tested and then choose the ones that gave the best results. In our case, starch was OK for many species.
For what species ? I am amazed that nobody ever gives relevant details for a question or an answer.
Price of gelling agent is differ within the country...in my country gelrite or phytogel is the most expensive compared to agar.raw agar from seaweed is the chipest one.but may need higher amount..it about RM5/kg.tq
Hi Gaber,
What agarose concentration do you use for rice? I am curious...
Try with Kithul ( Caryota urens ) Starch
PS: Kitul is a palm endemic to Sri Lanka's rain forests.
Mixtures of gelling agents can sometimes work well. For example, a starch/Gelrite mixture worked well for apple and raspberry micropropagation - quite a bit better than agar. They used 50 g/L corn starch + 0.5 g/L Gelrite. They estimated that this mixture was only 10-15% of the cost of agar. However, they test the effects of starch or Gelrite alone.
Zimmerman RH, Bhaardwaj SV, Fordham IM (1995) Use of starch-gelled medium for tissue culture of some fruit crops. Plant Cell Tiss. Org. Cult. 43: 207-213
Liquid medium with glass beeds
jack fruit pulp with half it normal moisture, supplemented with nutrients based on crop and cycle
I repeat, it depends on the species. tests must be done and economic calculations have to be done in case of positive results with certain experimental treatments.
It also depends on the region, country etc.
We are marching with wheat starch, here in Romania but in countries with other climates and other traditions and possibilities the situation may be diffeent regarding the cost and availability of gelling agents.
I suggest trying Psyllium husk, tapioca pearls, guar gum, gum katyra, gum arabic, carob locust gum, wheat, corn or potato starch and many others. This is a vast area of research and quite a new one. It is like discovering a new ocean. There are already many articles available on this topic
How do you use gelatin ? We tried gelatin and we also tried Pectin among many others and these two did not gel the media at any concentration.
We use jelly moss instead of agar. It proves to be good even for species such as Santalum album. Sago also found to be an alternative. Those things make low cost per plant in production
If by "Jelly Moss" you mean carrageenan, there are many carrageenans from different Genera and even different Kingdoms. For consistant results you have to use the same one such as GP-812.
Alexandru, could you possibly tell me how to use the alternatives like Tapioca pearls or gum katyra?
u can use isubghol or corn starch as well, they showed good results during my work
You may want to use alginate. You will find sodium alginate in the market, but it can be coverted to calcium alginate, wich is the gel form. Prepare a 1% solution of sodium alginate and pour it in dialisis bags, Then inmerse the bags in 10% Calcium Cloride solution over night and you wil get a gel the next day. Let me know the outcome. Good luck.
If you are interested in trying carrageenan like an alternative for agar in plant tissue culture, I can supply you with samples of pure grades. It's important to know if you want to obtain a liquid medium (a very low dose of lambda carrageenan could work) or a semisolid medium (where iota carrageenan should be the best option).
Cassava/tapioca - pearls/sago/sabudana has proven cost effective
https://goo.gl/3fwxpa https://goo.gl/8yjG3T https://goo.gl/ED8sBf
Dear Danial,
Sorry, sorry, I meant carrageenan. They are a very good option.
low cost gelling agent is very important, but sometimes, the results may vary due to different solidification agent,
Gelling agents provide support to the cultured plants in vitro. In routine, standard quality (purity) agar is the universal gelling agent used globally. To switch for the support matrix, PU foam in most economic for the tissue culture. Still, the varietal response may be variable or even leads to abnormal growth e.g.vitrification. The second alternate will be the gelrite, which is effective and being used as 1/3 (by weight) as compared to agar. The price may be more but it will be cost effective.
Danial Kahrizi please give me your email and telephone number details in order to send you the offered samples (they are requested by the courier service).
there are many options for that try cassava starch. but what about the temperature it is very important agent to detect which kind of gelling agent you want to use for which kind of work. Melting and solidification temp. are very importance for work successful.
Hi
As we all know, the best is agar but as a low cost choice confectiinary agar is an approperiate alternative.
you can also use some growth matrix just to support root stablishment. perlite is ok. but I never rcommand it for commercial scale do to it is labour-costly-alternative! !
If your plant can uptake nutrients and give a good response in the media, low-quality agar is ok in my sense. Since we discard the gel while transfer plants to other flask it is economically friendly.
Numerous articles have referred to agar alternatives such as starch, guar gum, gum katira, xanthan gum, Agar Agar, sago, agarose, gelrite, alginates, starch-gelrite mixture but we use in commercial level of the composition of edible Agar-Based with agar which is much cheaper, and we have not seen any problems.
Hi Danial, have you tried some of the suggested agar replacements?
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