I am working on Yam in vitro regeneration through meristem culture from field originated. About 70% meristem from any accession of this yam species takes relatively too long to regenerate into shoot and delayed of expected Virus Free in vitro conservation and distribution to partners. Attempts towards establishing yam in vitro genebank by meristem culture methods have not met much success. Some meristem accessions took a year longer to regenerate into shoot due to somaclone like calusing, browning, sprouting but not shooting, and eventually turn dark and died.
Hi,
Probably you can find some answer here:
Bömer, M., Rathnayake, A. I., Visendi, P., Sewe, S. O., Sicat, J. P. A., Silva, G., ... & Seal, S. E. (2019). Tissue culture and next-generation sequencing: A combined approach for detecting yam (Dioscorea spp.) viruses. Physiological and molecular plant pathology, 105, 54-66.
Hi Ricardo,
Thank you very much. It really helpful, expecially on the virus cleaning side.
You're welcome. Also search for the Bernard Malaurie's works.
Hi Ricardo, I checked Bernard Malaurie's work and were relevant to my work.
I appreciate your recommendation.
From this paper: "When the maize KN1 gene was constitutively overexpressed (35S::ZmKN1) in Nicotiana tabacum, it resulted in a 3-fold increase in shoot organogenesis, relative to the NPTII-only control [43]. " There are also other methods described in this paper to increase genetic transformation efficiency of some plant species which are difficult for genetically transformation.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6409764/pdf/plants-08-00038.pdf
You should wait for a long time when using meristem culture method, especially when the meristem size is less than 0.5 mm.
Hi Nguyen, thank your for your advice. That is actually one of my major concern when meristem culture regenerate very quickly and other side as you said, wait for a long time which I guess that it might be either due to hormonal imbalance, varietal different, somaclonal variation, chromosomal rearrangement and other physiological factors. There are two keys area concern: 1) We expect meristem culture to clean virus infected mother plant. 2) plantlet regeneration from direct organogenesis without undergoing embryonic stages. As you know, not all Breeders, Researchers or Requestors can wait for a long time to receive virus free materials for their breeding work program. I have carried out a lot of experiment to indentify an appropriate medium and combination of plant growth regulators to regenerate shoot from meristem culture on yam Dioscorea spp. But shoot regeneration success rate is very low and still remain bottle neck. So, I want to be guided more from all you expertise in the house on how to link this problem with scientific approach to conduct an experiment?
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