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Proceedings of Brainstorming meeting and Training cum demonstration on Cryopreservation and in vitro conservation in Horticultural Crops held at IIHR, Bangalore on 21st and 22nd February 14

Proceedings of Brainstorming meeting and Training cum demonstration on Cryopreservation and in vitro conservation in Horticultural Crops held at IIHR, Bangalore on 21st and 22nd February 14


Dr. N .K. Krishna Kumar, DDG-H addressing the delegates of the Brainstorming Meeting


The Society for Promotion of Horticulture Bangalore in collaboration with Indian Institute of Horticultural Research, Hessaraghatta, Bengaluru and Bioversity International organized a Brainstorming Meeting and Training cum Demonstration on Cryopreservation and in- vitro conservation in Horticultural Crop Genetic Resources on 21st and 22nd February 2014. The main objective of the program was to assess use of cryopreservation and tissue culture techniques for conservation and management of PGR in horticulture crops, enhance the use of in vitro conservation and cryopreservation protocols for horticulture crop genetic resources and equip participants with essential knowledge necessary for developing and using in vitro & cryopreservation techniques. The Brainstorm meeting primarily focussed on presentations by experts in the field, followed by discussions and interactions. The meeting was followed by training cum demonstration which included in depth analysis of basic research in the plant cryopreservation and in vitro conservation already developed by IIHR and other ICAR institutions under Horticulture SMD as well as reports on state-of-the-art methods, research and successes achieved with selected horticultural crops.


The inaugural session of the meeting was held in the IIHR Auditorium. Dr. S. Ganeshan, Organizing Secretary of the Meeting (Head & Principal scientist, Division of PGR, IIHR.) welcomed the Delegates, Dignitaries and Participants and gave a brief overview of two day program. Dr. C.K. Narayana, Director, IIHR delivered the opening remarks citing the importance of conservation of PGR in light of its erosion due to various kinds of human activities and natural causes. Dr. V.A. Parthasarathy, National Project Coordinator, UNEP-GEF Programme, Bioversity International reflected on the significance and importance of cryopreservation. He emphasized the need to use state of the art technologies for genetic resources conservation.


Dr N. K. Krishna Kumar, Patron SPH & DDG (Hort.), ICAR stressed upon the need to train human resources and use cryopreservation technology in horticultural crops, which would usher in higher levels of genetic resource availability to breeders thereby paving way for development and release of new varieties and hybrids. Referring to the Coorg Honey Dew papaya variety, he said that the pollen collections made during the 80s at IIHR are still available in the Cryobank for use by breeders,


The Chief Guest Padmashree Dr. K. L. Chadha (Formerly DDG-Hort.) ICAR, released the “Compendium on cryopreservation and in vitro conservation in Horticultural crops”. In his inaugural address, he traced the historical perspective as to how the program on cryopreservation of pollen was conceived at IIHR through an FAO-UNDP project in the early 80s under his guidance as Director, IIHR.


Dr. K. Madhavi Reddy, Secretary, SPH and Principal Scientist, Division of Vegetable crops, IIHR, proposed vote of thanks, on behalf of the organization Committee.


Technical Session- I


The first technical session was chaired by Dr V.A. Parthasarathy, co-chaired by Dr P.N. Krishnan while Dr L.K. Bharathi was the rapporteur.


There were four presentations in this session:


The first presentation was on “Cryobanking of plant germplasm at Cryogenebank at NBPGR” presented by Dr Rekha Choudhury, Head, Tissue Culture & Cryopreservation Unit, NBPGR, New Delhi. She explained the quantum of work being carried out at NBPGR on cryopreservation of various crops specifically on cryopreservation of seeds and dormant buds of temperate crops. She emphasized on giving more focus and future thrust of research for conservation of crops such as Garcinia, Citrus, Syzigium and many more, which are currently difficult to conserve. She also detailed the constraints related to long term conservation of seeds such as loss of seed viability in seed bank stored at -20oC and expression of incompatibility of cryopreserved buds in grafting. During interactions, the advantage of cryopreserving orthodox seed material and need for storing heterozygous seed material was also discussed.




  1. Prioritization of crops and centres for in vitro conservation.
  2. Optimization of cryopreservation protocols to conserve species like Garcinia, Syzigium and Citrus which are difficult to conserve in vitro.


Second presentation was made by Dr P.N. Krishnan, Head, Biotechnology, JNTBGRI, on the topic “In vitro and Cryopreservation research at JNTBGRI”. He described various activities of   in vitro cryopreservation of rare, endangered and threatened species at JNTBGRI. Long term conservation of shoot tips, seeds, embryos of RET species, pollen & seeds of orchid species (as synthetic seeds)
 was also presented by him.




  1. NRC Orchids, Sikkim and CPCRI Kasaragod can initiate collaboration with JNTBGRI, Thiruvananthapuram.


In the third presentation Dr K. Nirmal Babu, Coordinator-Spices, IISR, Kozhikode explained in detail the intricacies of cryopreservation of spices in his lecture on “In vitro conservation and Cryopreservation in spices”. He emphasized on reducing the evaporation and growth rate of plants in cryopreservation and importance of proper labeling and documentation. If such basic steps are followed meticulously, any number of accessions could be stored for a long time. He also emphasized on cell line, pollen and DNA banking for cryopreservation of rare material. He also mentioned that in case of vegetatively propagated spice crops like ginger or vanilla where seed setting is absent, the only means of conservation is through cryopreservation.





  1. Advancing technique of microrhizome production as a means of in vitro conservation

The last presentation of the session was made by Dr Anitha Karun, Head, Crop Improvement, CPCRI, on the "Present status and future prospects of palm genetic resource conservation by cryopreservation”. She made a detailed presentation on the embryo culture protocols developed in coconut and tissue culture protocols of areca nut and oil palm. She highlighted the importance of coconut plumule culture where all plants are genetically uniform. She also emphasized the need for initial drying/desiccation of pollen before cryopreservation is carried out.



  1. Protocols developed for coconut cryopreservation may also be extended to areca nut and oil palm for cryopreservation



Technical Session- II


The Technical Session- II was chaired by Dr. P.S.Naik, Director, Indian Institute of Vegetable Research, Varanasi and Co-Chaired by Dr. Rekha Choudhary Head, Tissue-culture and Cryopresevation at NBPGR, New Delhi.


There were five presentations in this session


First presentation was on “In- vitro conservation of potato germplasm experiences at CPRI, Shimla”, by Dr. Jai Gopal, Director, NRCOG. In his presentation the achievements related to conservation of potato grmplasm made by CPRI was explained. In general the cultivars are maintained through vegetative propagation, in order to maintain their genetic purity. The major constraint in field genebank conservation is requirement of large area and therefore alternatively elite parental lines are conserved as in vitro propagated micro plantlets under disease free culture conditions. Dr. Jai Gopal also explained the different micropropagation techniques optimized at CPRI Shimla. Using this method plants can be conserved for 2-3 years. Plantlet maintained at low temperature is ideal in conservation of potato genetic resource. Alternative method is production of micro tubers for storage up to one year.



  1. The micro tuber propagation technique can be adopted in other tuber crops for multiplication and conservation.


The next presentation was on “Cryopreservation of Genetic Resources in Potato” by Dr. P.S.Naik, Director, IIVR, Varanasi. He explained the traditional methods for conservation of Potato genetic resources. He also mentioned the advantages of in vitro conservation of potato over traditional methods viz. medium and long term conservation. He further explained the important steps and care to be taken while going for cryopreservation of Potato and also emphasized on the various issues related to cryopreservation.




  1. Linkage and co-ordination is required between the institutions involved in vitro and cryo-preservation of agricultural crops.
  2. It is necessary to prioritize the crops before going for Cryopreservation.


The third presentation was on “Pollen cryo banking- implication in genetic conservation and plant breeding” which was presented by Dr. S. Ganeshan, Head, PGR IIHR, Bangalore. Dr. Ganeshan explained the different levels of conservation. He detailed the advantages and need of pollen cryo-preservation with appropriate examples. He also presented the methodology to be followed for collection of pollen for cryopreservation and explained various uses of cryo preserved pollen. He also explained various post storage viability tests. He briefed the house on the pollen cryopreservation activities presently being carried out at IIHR.


The next presentation was on “In -vitro conservation in Horticultural Crops” presented by  Dr.P.E. Rajasekharan, Principal Scientist, IIHR, Bangalore. He gave an overview of the in vitro multiplication and conservation methodologies carried out at IIHR where in they have also taken care of conserving the RET medicinal plants. He also mentioned the pros and cons of in vitro conservation and explained the protocols developed for in vitro multiplication of RET plants.


The last presentation was on “Banana Conservation strategies” presented by Dr. Saraswathi, from NRC Banana. She presented an overview of the germplasm accessions conserved at various research organizations in India. She informed that 1603 accessions are maintained. She mentioned that though FGB is easy method of conservation of banana but it is commonly affected by diseases and natural calamities. She also briefed on the in vitro conservation activities going on at NRCB, Trichi.



Seeds of wild species of banana to be conserved by cryopreservation.


Crops prioritized for further action

 Tissue/ organ   Conservation


Cryopreservation                                                       Researchable crops for developing protocols

Papaya                                                                                                 Mango

Guava                                                                                                  Garcinia

Coconut                                                                                               Date palm

Citrus                                                                                                   Nutmeg

Banana                                                                                                Litchi

Areca nut                                                                                             Ber

Sweet Potato                                                                                        Jamun

Potato                                                                                                  Cherry

Black Pepper                                                                                       Garlic

Cardamom                                                                                           Oil palm

Turmeric                                                                                              Amla

Ginger                                                                                                  Moringa

Cassava                                                                                               Annona muricata





Jack Fruit



Aloe vera

Chlorophytum spp.

Piper logum


Pointed gourd


                             Plenary session in progress


Training cum demonstration on Cryopreservation and in vitro conservation in Horticultural Crops:


For the benefit of the participants of the brains storming meeting , a practical session on pollen cryopreservation and in vitro conservation was conducted on 22nd February 14 in the Pollen storage laboratory of Division of Plant Genetic Resources. In the beginning the participants were taken to RET field gene bank for showing the flowering and collection of pollen from field. Following these they were brought to the laboratory and shown how to extract pollen and do the viability assessment using various methods (hanging drop and cellophane).  The requirements for the same were also explained. The trainees were asked to do the viability assessment o the extracted pollen. Following this they were taken to the facilities and explained how cryopreservation is done. All these were done in the morning. All these were carried out under the leadership of  Dr. P. E. Rajasekharan Principal Scientist division of PGR.



Participants in field learning pollen collection and back in the laboratory involved in germinating pollen


In the afternoon the in vitro conservation practical exercises were conducted and trainees were also shown  the facilities. Dr. Anuradha Sane explained how genetic fidelity test were done for tissue cultured plants




Updated on 05.03.2014