Hello, I am not familiar with this problematic so far.
However, I am interested in rapid and precise method, how to recognize the most responsive genotype of pea (even wild peas) to Rhizobium and other inoculants.
Is there any published LABORATORY method how to select out the best "partners"? I know the best result should be taken in field, however, this is the first step. Sorry, if my question is not satisfactorily clear...
Thanks, Milan
Hi Milan,
SORRY! I don't know much about plant response to Rhizobium and other inoculants.
Fugui
Hello Dear milan
regard to the growth promoting power of Rhizobium to the plant i think you should notice the the growth factor of plant.
Dear milan,
In my opinion, First of all, you should focus on the number of genotypes of pea as well as the number of Rhizobium strains that you want to work on it.
This will generate a high number of trials. once you have this data, you should carry out pots trials for each pea genotype VS each Rhizobium strains. with replicates of course.
the parameters that you can follow in order to measure the response are :
the length of shoots and roots.
the dry weight of roots and shoots.
The number of nodules/color of nodules(pink or white): the pink ones have a high nitrogen fixation activity.
Measure the concentration of nitrogen(AMMONIUM) in leaves.
the last parameter is the ARA test: is the most accurate test for nitrogen fixation activity but a little bit complicated and requires Gc-MS equipment and some gazes such as ethylene and acetylene.
Good luck Dear Milan,
I wish you the best,
Dear Dr. Milan, according to my opinion you should go for qualitative and quantitative estimation of different growth promoting traits of rhizobium and based on these estimation you should select the most potential strain. Thereafter, you should go for inoculation in the desired genotypes of your plant.
Regarding this you can follow the paper of Marques et al, 2010. And the attached file would also help you to some extent.
Regards,
Sosanka
Article Phosphorus Solubilization and Plant Growth Promotion Ability...
Thank Sosanka, just one more question - where can I get some information about "qualitative and quantitative estimation of .....promoting traits..."? Is somewhere some kind of database. And if i will chose some strain, is there any recommendation what is the best way how to inoculate seeds? (I am really not familiar with this topic). Or if this information is is Marques et al., 2013, can you please send me more detailed specification of this paper?
Thanks again
Okk. I am attaching the publications for your needfull.
Best of luck!
You can go for pot experiments with the various strains vs the genotypes to determine the strains and genotypes that fix the highest amount of nitrogen. You can also asses root and shoot biomass as well as nodule number and dry weight. These are good indicators to be assessed for symbiotic efficiecy
Thanks Muhammed, I can imagine I have several strains. However, in pot experiments, is there any rule, how "much" inoculant I have to give to each pot? what should be the pH for different kind of soils? And, in the beginning, I was asking for LABORATORY method, not for pot experiments....Anyway, thanks
The best reference, Book:
Somasegaran, P and Hoben, H.J.
Methods in Legume-Rhizobium Technology. , NifTAL, Univ. Hawaii, and
Same authors:
Handbook of Rhizobia. 1994.
Springer.
Dear Milan,
All the answers above need to be taken into consideration in regard to your goals. I simply had few practical comments.
In your experiment, you have two partners; the plant and the rhizobium. Their symbiosis is under biochemical communications that allow or stop the nodule development. For one strain of rhizobium, you may find some genotypes that produce too much nodules to too few nodules. A different strain of rhizobium will not give the same results.
You need to standardize your process. Planting seed in growth pouches is very useful, even for large seeds. Also, pot experiments can be selected. You need a substrate that can be repeated over time (ex. : mixture of perlite, vermiculite, etc.). Your inoculum should be growth on Yeast Manitol Medium; then harvest it and dilute it to about 100 000 000 rhizobium per ml. You can prepare it in advance and keep it at 4 C. You have to water your plant with N free nutritive solution to maximize seedling emergence and rhizobium survival. At seeding, simply place 1 ml of inoculum per seed. You can do a second inoculation one week later. Often, it is after the development of the first true leaf that nodules start to develop. Nodulation is checked 3-4 weeks after seed germination and is very good at early bloom.
If you have a lot of genotypes and a lot of rhizobial strains, keep your harvest simple. Rhizobium should fixes nitrogen and the plant should have a better growth. So, the plant weights (dry and fresh weights) should give your answer.
Good luck and best regards,
Dear prof. Beauchamp, thanks fore detailed answer, which finally give me some basic for the laboratory-based method. Of course, I will also do a field (or pot) experiment, however, the pre-selection is always necessary...
Milan
Dear Milan,
Your goal appears to select a method for rhizobium inoculation for pea genotype selections. I will suggest you to test your best commercial rhizobium strains with your best pea cultivars. [For more information on rhizobium inoculation, please read -Methods for Testing Legume Inoculant on line at : http://www.inspection.gc.ca/plants/fertilizers/sampling-and-testing/methods-for-testing/eng/1308033966900/1308034369347).] You will find the best «strain-pea» combination that give the best yields under your climatic conditions. This will decrease the number of strains to be check. Your « Positive Control » will be the best strain with the best cultivar that you will use to compare the yields of your pea genotypes. Your negative control will be the best cultivar well fertilized with mineral nitrogen.
All the best,
Dear all,
For the same experiment with using sand and peat, should I sterilize the soil or can i work with it without sterilization?
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