Innovative Cryopreservation Techniques and Fire Blight Management in Apple Collections

The provided content discusses cryopreservation techniques for Malus and Prunus collections, focusing on bud segments and injury management. Measures to minimize fire blight in the PGRU apple collection are highlighted, including cryogenic storage of buds and repropagation on specific rootstocks. A pilot project on the recovery and longevity of cryopreserved apple buds over several years is also detailed, showing promising results in bud recovery after storage in liquid nitrogen. The overall emphasis is on conservation and preservation strategies for apple genetic resources.

• Cryopreservation
• Apple Collections
• Fire Blight Management
• Genetic Resources
• Bud Recovery

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1. CRYOPRESERVATION OF MALUS A BACK UP COLLECTION AT $1/ACCESSION/YR! Base collection (2146 acc.) at NCGRP, Ft. Collins, CO Processing 35 mm bud segments Hundreds of seedlings budded with cryopreserved buds testing viability Recovery of accessions by bud grafting Active collection (436 acc.) at PGRU Geneva, NY

2. CRYOPRESERVATION OF PRUNUS Base collection (52 acc.) at NCGRP, Ft. Collins, CO Injury to primary bud following LN exposure Active collection (8 acc.) at PGRU Geneva, NY Typical recovery scenario from axillary bud

3. MEASURES BEING USED TO MINIMIZE FIRE BLIGHT IN THE PGRU APPLE COLLECTION Cryogenic storage of dormant buds; as a result, ~ 90 accessions that have died (fire blight) have been rescued by direct grafting of cryopreserved buds Repropagation of collection to EMLA 7 rootstock: Replacing collection previously held on seedling and EMLA 9 rootstock Application of Apogee post-bloom to control vegetative growth reducing shoot blight Removal of all lesions in dormant season; timely annual copper spray; and corrective pruning of infected shoots throughout the growing season Continued use of antibiotics under defined protocols to minimize blossom blight

4. PILOT PROJECT: FORSLINE, P.L., C. STUSHNOFF, L.E. TOWILL, J.W. WADDELL, W.F. LAMBOY AND J. R. MCFERSON. 1998. RECOVERY AND LONGEVITY OF CRYOPRESERVED APPLE BUDS. J. AMER. SOC. HORT. SCI. 123:365-370. 84 accessions were processed periodically over 4-yr period 12/12/1988 8 accessions 02/07/1989 8 accessions 12/14/1990 12 accessions 01/15/1991 13 accessions 12/12/1989 6 accessions 01/19/1990 9 accessions 12/18/1991 13 accessions 01/18/1992 15 accessions

5. Pilot project to determine protocol for cryogenic storage of dormant buds: % bud recovery of 84 apple accessions (processed 1989-1992); tested after up to 8 years of storage in liquid nitrogen Recovery % Treatment Desiccated Control 85.3 aZ Storage one month 63.0 b Storage one year 64.2 b Storage two years 66.5 b Storage four years 68.6 b Storage after eightY years 68.3 b ZSeparation of grand means of 84 accessions at P < 0.01 by test for differences between two proportions (LSD = 7.2) Y Fifteen year testin process 2004 to 2007

6. ANNUAL CRYOPRESERVATIONOF MALUSACCESSIONS AT NCGRP FOLLOWING THE 4-YR PILOT PROJECT 1988-1992 M. x domestica 86 108 167 169 223 209 120 108 52 Year 1993 1994 1995 1996 1997 1998 1999 2000 2003 Hybrids 8 7 25 24 71 66 55 46 6 Other species 5 26 50 49 70 64 71 84 56 Total 99 141 242 242 364 339 246 238 114 2004 Total 22 1264 2 97 572 121 2146 310

7. SUCCESSFUL CRYOPRESEVATION FOR > 90% OF ACCESSIONS STORED AT NCGRP: THOSE WITH < 30% VIABILITY WILL BE REPROCESSED 1760 or 91% were successful 700 171 or 9% of total were unsuccessful 600 500 0-10 % 20-30 % 40-50 % 60-70 % 80-90 % 100% No. of accessions in 400 300 200 storage 0-10 20-30 40-50 60-70 80-90 100 100 0 Categories of success based on bud recovery via grafting

8. SUCCESSFUL CRYOPRESEVATION FOR > 95% OF ACCESSIONS STORED AT PGRU: THOSE WITH < 30% VIABILITY WILL BE REPROCESSED 418 or 96% were successful 180 18 or 4% of total were unsuccessful No. of accessions in storage 160 140 0-10 % 20-30 % 40-50 % 60-70 % 80-90 % 100% 120 100 80 60 40 20 0-10 20-30 40-50 60-70 80-90 100 0 Categories of success based on bud recovery via grafting