Using different pre-germination treatments, maximum response and plant establishment was obtained by treating the seeds with water (45°C) for 90 seconds. Experiments were laid on domestication studies and impact of hot water treatments on the subsequent growth. 

The endangered A. heterophyllum was successfully conserved and domesticated under ex situ conditions (greenhouse and open). Water treatment at 45° and 50° C for 90 seconds not only yielded high seed germination but also improved the growth of the plants over the control. This was evident from the correlation studies which revealed not only enhanced vegetative growth but also better reproductive yield (Fig. 5.5). Total available leaf area/ plant played a major role in the production of flowers and fruits rather than seeds. A significantly higher correlation between the number of flowers and fruits/plant than with seeds/fruit and number/mass (Fig. 5.6) indicated that investments were made towards production of fruits, yet a large percentage of the seeds underwent abortion.  

Germination of seeds was initiated during March-April under natural conditions. However, under ex situ, the germination and early seedling growth was in October-November and hence the duration and events of life cycle got modified and the seeds matured during late May-June (Fig. 5.7).    

The average biomass was also evaluated in plants germinated and established under greenhouse. 7.77 g and 9.56 g/ plant yield (fresh) were in 1 year and 2 years old respectively. Alkaloid content of the dried roots/rhizomes was 1.67 and 2.4 % in 1 and 2 years old plants respectively and was almost at par (2.9%) present in the rhizomes from Chamba. This is an important step towards the sustainable commercialization.

SHAPE \* MERGEFORMAT

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CRYO-PRESERVATION

Experiments were initiated on cryo-preservation of A. heterophyllum and P. hexandrum seeds collected in the year. In A. heterophyllum,the initial moisture percentage was low (6%) and were therefore directly stored under liquid nitrogen. The seeds were retrieved after regular intervals for evaluation of viability, germination and cryo injury. Seeds showed about 90-100 % germination after 30 days storage. However, there was a higher ion leakage in the seeds stored for 30 days as compared to the seeds stored for 10 days only. The seeds of Podophyllum had much higher initial moisture content (50%) and were therefore initially desiccated to 10 and 5 moisture levels and then stored in liquid nitrogen. Preliminary results showed that seeds stored with 10% moisture had only 50% viability as compared to 70% in the seeds stored with 5% moisture levels for 10 days. Further time interval studies and development of protocols for liquid nitrogen storage were undertaken. 

Fig. 5.7 Life cycle of Aconitum heterophyllum under ex situ at Palampur