try to find a solution in this paper
http://ipps.org/Papers/NewZealand/seelye.pdf
It is well documented in the previous literature that, when in vitro mango plantlets get transferred to the autotrophic mode i.e. transfer to the soil, the reason for less survival or difficulty in hardening is due to divergent or different leaf anatomy or morphology from the normal grown plants, malfunctioning of stomata, poor development of cuticular wax and of palisade tissue and with low photosynthetic efficiency mainly due to high humidity (90%) and sugar containing medium obtained in in vitro. Thus, they are amenable to transplantation shock.
Regards
Shiwani Kaushal
Dear Rajesh,
Nothing is tough. Stop taking too many precautions. Try to root shoots in half to one-fourth strength MS/B5 or any other medium you are using and in case you are getting plantlets directly with both root and shoot then transfer them to half to one forth strength medium. Or else, after getting well rooted plantlets, transfer them into medium which is half to one fourth strength, please remove sucrose or any other sugar your are using and all organics at the time of rooting and/or transfer after plantlets develop healthy root system. Another, important aspect you need to remembers is regarding the size of the plantlets - we prefer transferring well rooted shoot/plantlet with shoot length of 4-5 cm). Ones you have plantlets acclimatized to such conditions, please wash the roots of each plantlet well with tap water (if your plantlets are healthy, there would be no problem in washing with tap water). Simply plant these plantlets in a manner similar to what we stated in our papers (with such simple protocols, we have been successful in establishing even tougher systems like chickpea with ease - you can get 100% success).
Please download Prasad et al. and Firoz et al. papers from our ResearchGate site.
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