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ARS Home » Pacific West Area » Corvallis, Oregon » National Clonal Germplasm Repository » Research » Publications at this Location » Publication #223230
Title: Medium Container and Genotype All Influence In Vitro Cold Storage of Apple Germplasm

Author
item KOVALCHUK, IRINA - INSTITUTE PLANT BIOLOGY
item LYUDVIKOVAL, YELENA - INSTITUTE PLANT BIOLOGY
item VOLGENIAL, MARIAM - INSTITUTE PLANT BIOLOGY
item Reed, Barbara

Submitted to: Plant Cell Tissue and Organ Culture
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/5/2008
Publication Date: 1/20/2009
Citation: Kovalchuk, I., Lyudvikoval, Y., Volgenial, M., Reed, B.M. 2009. Medium Container and Genotype All Influence In Vitro Cold Storage of Apple Germplasm. Plant Cell Tissue And Organ Culture. 96:127-136.

Interpretive Summary: The goal of this study was to evaluate tissue culture cold storage of apple by screening a range of diverse types and with more comprehensive testing of multiple parameters on two genotypes, cultivar ‘Grushovka Vernenskaya’ and wild selection TM-6. Stored plants were rated on a 6 point scale (0 low to 5 high) for plant appearance at 3 mo intervals after storage at 4 °C. Combinations of growth factors were studied in three types of containers (tissue culture bags, test tubes or jars). An initial screen of 16 genotypes stored in tissue culture bags indicated that plantlets could be stored at 4 °C for 9-14 months without subculture on standard medium with no plant growth regulators (PGRs). In subsequent studies on the two selected genotypes, the medium, container and genotype were all important factors in the length of storage. Overall it appears that cold storage of apple shoot cultures can be successful for 21 months in tissue culture bags with 25% MS nitrogen, 3% sucrose, and no PGRs or for 33 months in jars or tubes on MS with 3% sucrose and PGRs. Genetic analysis found no significant differences among plants stored for 39 months and non-stored controls.

Technical Abstract: The goal of this study was to evaluate the in vitro storage of apple germplasm by screening a range of genotypes and with more comprehensive testing of multiple parameters on two Kazakhstan genotypes, Malus domestica L. cultivar ‘Grushovka Vernenskaya’ and wild selection Malus sieversii (Ledeb.) M. Roem,TM-6. Stored plants were rated on a 6 point scale (0 low to 5 high) for plant appearance at 3 mo intervals after storage at 4 °C. Combinations of carbon source (sucrose and/or mannitol), nitrogen content (25, 50 or 100%) and plant growth regulators (ABA, BAP, IBA) were studied in three types of containers (tissue culture bags, test tubes or jars). An initial screen of 16 genotypes stored in tissue culture bags indicated that plantlets could be stored at 4 °C for 9-14 months without subculture on standard 3% sucrose Murashige and Skoog (1962) (MS) medium with no plant growth regulators (PGRs). In subsequent studies on the two selected genotypes, ANOVA indicated highly significant interactions among medium, container and genotype. ‘Grushovka Vernenskaya’ shoots on MS with no PGRs and 3% sucrose remained viable (ratings of =1) for 21 months of storage in bags. Storage on reduced nitrogen (MS with 25% nitrogen), PGRs, and 3% sucrose kept ‘Grushovka Vernenskaya’ shoot condition rated >2 at 21 months. Addition of 0.5 or 1 mg-1 abscisic acid (ABA) also improved plant ratings at 21 months. The longest storage for ‘Grushovka Vernenskaya’ was 33-39 months on MS with PGRs and 3% sucrose in either tubes or jars. Addition of abscisic acid (ABA) to the medium did not improve storage of plantlets in jars and tubes at 15 months. A wild-selected Kazakhstan genotype TM-6 stored best in tubes on 3% sucrose with PGRs or in jars on 2% mannitol and 2% sucrose. Overall it appears that cold storage of apple shoot cultures can be successful for 21 months in tissue culture bags with 25% MS nitrogen, 3% sucrose, and no PGRs or for 33 months in jars or tubes on MS with 3% sucrose and PGRs. RAPD analysis found no significant differences among plants stored for 39 months and non-stored controls.