IN VITRO CULTURE AND CRYOPRESERVATION PROGRAM

 

PROGRAM GOALS: The research encompasses four related areas:

 

1.       Development of improved and broadly applicable methods for tissue culture of crop germplasm at the repository.  The specific objective is the identification of media, growth regulators, and other growth factors for the culture of a range of genotypes within each genus represented at NCGR, Corvallis.

 

2.       Development of low temperature or other slow growth storage techniques for in vitro cultures.  The specific objective is to provide conditions for in vitro backup of the germplasm collections that can also be used throughout the world germplasm community.  A cold storage collection is monitored to collect data on the storage life of individual accessions in culture.  Alternative methods of room temperature storage are explored.

 

 

3.       Development of cryopreservation methods to facilitate the future storage of germplasm in liquid nitrogen.  Experiments are performed to evaluate pretreatment techniques, cryoprotectants, freezing protocols and recovery stages.  New techniques and protocols are developed or adapted for the range of plant response.  Germplasm screening determines the applicability of a specific technique to a broad range of genotypes.  Technology transfer of cryogenic techniques to other genetic resources labs.

 

4.       Evaluation of the genetic stability of in vitro-stored plants and cryopreserved meristems.

 

PUBLICATIONS:

 

IN VITRO CULTURE

1.      Reed, B. M.  Multiplication of Rubus germplasm in vitro: A screen of 256 accessions.  Fruit Varieties Journal 44(3):141-148.  1990.

2.      Reed, B.M.  Micropropagation of meadowfoam (Limnanthes spp).  Plant Cell Reports.  10:94-96. 1991.

3.      Reed, B.M. and A. Abdelnour-Esquivel.  The use of zeatin to initiate in vitro cultures of Vaccinium species and cultivars.  HortScience.  26(10):1320-1322.  1991.

4.      Yu, X. and B.M. Reed.  A micropropagation system for hazelnuts (Corylus species).  HortScience 30:120-123.  1995.

5.      Yeo, D. Y. and B.M. Reed.  Micropropagation of Pyrus rootstocks.  HortScience 30:620-623.  1995.

6.      Reed, B.M.  In vitro rooting response of Pyrus germplasm.  HortScience 30: 1392-1294.  1995.

7.      Tsao, C.W.V., J. D. Postman, and B.M. Reed.  Virus infections reduce in vitro multiplication of ‘Malling Landmark’ raspberry.  In Vitro Cellular and Developmental Biology – Plant 36:65-68.  1999.

8.      Bell, R. L. and B. M. Reed. In vitro Tissue Culture of Pear: Advances in Techniques for Micropropagation and Germplasm Preservation.  International Pear Congress 2000.  Acta Horticulturae.  596: 412-418.  2002.

9.      Reed, B.M. and K.E. Hummer. Genetic Stability of Strawberries in Culture, p. 98-101.  In: S.C. Hokanson and A.R. Jamieson (eds.).  Strawberry Research to 2001. ASHS Press, Alexandria, VA.  (Proceedings of the 5th North American Strawberry Conference) 2002.

10.  Tsao, C.W.V. and B.M. Reed.  Gelling agents, silver nitrate, and sequestrene iron influence adventitious shoot and callus production from Rubus Leaves.  In Vitro Cellular and Developmental Biology – Plant 38:29-32.  2002

 

IN VITRO CONTAMINATION

1.      Reed, B.M., P.M. Buckley and T.N. DeWilde.  Detection and eradication of endophytic bacteria from micropropagated mint plants.  In Vitro Cellular and Developmental Biology - Plant 31P:53-57.  1995.

2.      Buckley, P.M., T.N. DeWilde and B.M. Reed.  Characterization and identification of bacteria isolated from micropropagated mint plants.  In Vitro Cellular and Developmental Biology 31P:58-64.  1995.

3.      Reed, B.M. and P. Tanprasert.  Detection and control of bacterial contaminants of plant tissue cultures.  A review of recent literature.  Plant Tissue Culture and Biotechnology.  1:137-142 1995.

4.      Tanprasert, P. and B.M. Reed.  Detection and identification of bacterial contaminants from strawberry runner explants.  In Vitro Cellular and Developmental Biology - Plant 33:221-226.  1997.

5.      Tanprasert, P. and B.M. Reed.  Determination of minimal bactericidal and effective antibiotic treatment concentrations for bacterial contaminants from micropropagated strawberries.  In Vitro Cellular and Developmental Biology - Plant 33:227-230.  1997.

6.      Reed, B.M., J. Mentzer, P. Tanprasert, and X. Yu.  Internal bacterial contamination of micropropagated hazelnut:  Identification and antibiotic treatment.  In: Pathogen and Microbial Contamination management in Micropropagation   (Proceedings of the Second international Symposium on Bacteria and Bacteria-like Contaminants of Plant Tissue Cultures).  1997. pp. 171-176. Tanprasert, P. and B.M. Reed.  Detection and identification of bacterial contaminants of strawberry runner explants.  In: Pathogen and Microbial Contamination management in Micropropagation (Proceedings of the Second international Symposium on Bacteria and Bacteria-like Contaminants of Plant Tissue Cultures).  1997. pp. 141-145. 

7.      Tanprasert, P. and B.M. Reed.  Detection and identification of bacterial contaminants of strawberry runner explants.  Plant Cell, Tissue and Organ Culture 52: 53-55.  1998.

8.      Reed, B.M., Mentzer, J., Tanprasert, P., and X. Yu.  Internal bacterial contamination of micropropagated hazelnut: identification and antibiotic treatment.  Plant Cell, Tissue and Organ Culture 52: 67-70.  1998.

9.      Reed, B.M. and M.A.L. Smith (Eds).  Designing a Micropropagation System.  In Vitro Cellular and Developmental Biology – Plant 35: 275-284.1999.

10.  Reed, B.M. and P.M. Buckley.  Tissue Culture Contaminants Handbook.  USDA-ARS, Corvallis, OR 1999.  (Lab manual)

11.  Reed, B.M., Tsao, C.V., and C.L. Paynter.  (Eds) Lab Manual for the Tissue Culture Lab at the National Clonal Germplasm Repository, USDA-ARS, Corvallis, Oregon.  1999.  (Lab manual)

12.  Wang, N. and B.M. Reed. Development, Detection, and Elimination of Verticillium dahliae in Mint Shoot Cultures.  HortScience.  38:67-70. 2002.

 

 

COLD STORAGE OF IN VITRO CULTURES

1.      Reed, B.M.  Application of gas-permeable bags for in vitro cold storage of strawberry germplasm.  Plant Cell Reports 10:431-434.  1991.

2.      Reed, B.M. Cold storage of strawberries in vitro: A comparison of three storage systems.  Fruit Varieties Journal 46(2):98-102.  1992.

3.      Reed, B.M.  Improved survival of in vitro stored Rubus germplasm.  J. Amer. Soc. Hort. Sci. 118:890-895.  1993.

4.      Kumar, M.K., Barker, R.E., and Reed, B.M.  Morphological and molecular analysis of genetic stability in micropropagated Fragaria X ananassa cv. Pocahontas.  In Vitro Cellular and Developmental Biology - Plant  35: 254-258.  1999.

5.      Reed, B.M.  The In vitro genebank of temperate fruit and nut crops at the National Clonal Germplasm Repository-Corvallis.  In: F. Engelmann (ed).  Management of Field and In Vitro germplasm collections.  Proceedings of a Consultation Meeting, 15-20 January 1996,CIAT, Cali Columbia.  International Plant Genetic resources Institute, Rome, Italy.  1999.

6.      Hummer, K.E. and Reed, B.M.  Establishment and operation of a temperate clonal field genebank.  In: F. Engelmann (ed).  Management of Field and In Vitro germplasm collections.  Proceedings of a Consultation Meeting, 15-20 January 1996,CIAT, Cali Columbia.  International Plant Genetic resources Institute, Rome, Italy.  1999.

7.      Reed, B.M.  In vitro conservation of temperate tree fruit and nut crops.  In:  Plant Conservation Biotechnology pp. 139-154.  (Ed. E.E. Benson).  Taylor and Francis, London.  July 1999. 

8.      Reed, B.M.  In vitro storage conditions for mint germplasm.  HortScience 34:250-252.  1999.

9.      Reed, B.M.  The Basics of In Vitro Storage and Cryopreservation.  USDA-ARS, Corvallis, OR 1999.  (Lab manual)

10.  Reed, B.M. and K.E. Hummer.  Conservation of Germplasm of Strawberry (Fragaria Species) In: Y.P.S. Bajaj (Ed.) Biotechnology in Agriculture and Forestry Vol. 32.  pp. 354-370.  1995

11.  Reed, B.M., Paynter, C.L., DeNoma, J., and Y. Chang.  Techniques for medium- and long-term storage of pear (Pyrus sp.) genetic resources.  IPGRI Plant Genetic Resources Newsletter (peer reviewed).  15:1-5. 1998.

12.  Reed, B.M. Photoperiod Improves Long-Term Survival of In-Vitro Stored Strawberries.  HortScience. 37: 811-814. 2002.

 

 

CRYOPRESERVATION

1.      Reed, B. M. and H. B. Lagerstedt.  Freeze preservation of apical meristems of Rubus in liquid nitrogen.  HortScience.  22(2):302-303.  1987.

2.      Reed, B. M.  Cold acclimation as a method to improve survival of cryopreserved Rubus meristems.  Cryo-Letters.  9:166-171. 1988.

3.      Reed, B.M.  The effect of cold hardening and cooling rate on the survival of apical meristems of Vaccinium species frozen in liquid nitrogen.  Cryo-Letters 10:315-322.  1989.

4.      Reed, B. M. Survival of in vitro-grown apical meristems of Pyrus following cryopreservation.  HortScience 25(1):111-113.  1990.

5.      Reed, B.M. Germplasm cryopreservation:  Dependent on genotype or technique?  V Jornadas: Criopreservacion de Tejidos, Proceedings of the Spanish Society for Cryobiology.  November 1992.  pp 4-5 (In Spanish).  (Proceedings)

6.      Yu, X.L. and B.M. Reed.  Improved shoot multiplication of mature hazelnut (Corylus avellana L.) in vitro using glucose as a carbon source.  Plant Cell Reports 12:256-259.  1993.

7.      Reed, B.M.  Responses to ABA and cold acclimation are genotype dependent for cryopreserved blackberry and raspberry meristems.  Cryobiology 30:179-184.  1993.

8.      Reed, B.M.  The importance of pretreatment in the cryopreservation of plant tissues.  VI Jornadas Criopreservacion de Tejidos, Proceedings of the Interdisciplinary Spanish Society for Cryobiology Meeting November 1993.  pp 12-15.  (In Spanish).  (Proceedings)

9.      Normah, M.N., B.M. Reed and X. Yu.  Seed storage and cryopreservation behavior in hazelnut (Corylus avellana L. cv. Barcelona).  Cryo-Letters 15:315-322.  1994.

10.  Reed, B.M., M.N. Normah and X. Yu.  Stratification is necessary for successful cryopreservation of axes from stored hazelnut seed.  Cryo-Letters 15: 377-384.  1994.

11.  Reed, B.M. Cryopreservation of Seed:  Problems with some seed types.  VII Jornadas Criopreservacion de Tejidos, Proceedings of the Interdisciplinary Spanish Society for Cryobiology Meeting.  October 1994.  pp. 7-9 (In Spanish).  (Proceedings)

12.  Reed, B.M. and X. Yu.  Cryopreservation of in vitro-grown gooseberry and currant meristems.  Cryo-Letters 16:131-136.  1995.

13.  Reed, B.M.  The role of protein in the cryopreservation of plant cells and tissues.  VIII Jornadas Criopreservacion de Tejidos, Proceedings of the Spanish Society for Cryobiology, November 1995.  (In Spanish).  (Proceedings)

14.  Benson, E.E., B.M. Reed, R.M. Brennan, K.A. Clacher and D. A. Ross.  Use of Thermal analysis in the evaluation of cryopreservation protocols for Ribes nigrum L. germplasm.  Cryo-Letters 17:347-362.  1996.

15.  Reed, B.M.  Cryopreserved plant genebanks:  A dream come true.  IX Jornadas Criopreservacion de Tejidos, Proceedings of the Spanish Society for Cryobiology, November 1996.  (In Spanish).  (Proceedings)

16.  Reed, B.M. Chapter 3.  Pretreatment strategies for the cryopreservation of plant tissues.  In:  In vitro Conservation of Plant Genetic Resources.  (Eds.  MN Normah, MK Narimah, MM Clyde).  Plant Biotechnology laboratory, Faculty of Life Sciences, Universiti Kebangsaan, Malaysia.  1996.

17.  Luo, J. and B. M. Reed.  Abscisic acid-responsive protein, bovine serum albumin, and proline pretreatments improve recovery of in vitro currant shoot-tip meristems and callus cryopreserved by vitrification.  Cryobiology 34:240-250.  1997.

18.  Reed, B.M and Y. Chang.  Chapter 4.  Medium and long-term storage of in vitro cultures of temperate fruit and nut crops.  In: Conservation of Plant Genetic Resources In vitro.  Vol. 1 pp. 67-105.  (Eds.  M. K. Razdan and E. C. Cocking) M/S Science Publishers, Inc., USA 1997.

19.  Reed, B.M., DeNoma, J., Luo, J., Chang, Y., and L.E. Towill.  Cryopreservation and long-term storage of pear (Pyrus L.) genetic resources.  In vitro Cellular and Developmental Biology - Plant 34: 256-260.  1998.

20.  Chang, C. and B.M. Reed.  Extended cold acclimation and recovery medium alteration improve regrowth of Rubus shoot tips following cryopreservation.  Cryo-Letters 20:371-376.  1999.

21.  Chang Y. and B. M. Reed.  Extended Alternating-Temperature Cold Acclimation and Culture Duration Improve Pear Shoot Cryopreservation Cryobiology 40:311-322.  2000.

  1. Chang Y. and B.M. Reed.  Cold acclimation improves the cryopreservation of in vitro-grown Pyrus and Rubus meristems.  In: Cryopreservation of Tropical Germplasm.  Current research progress and application, Engelmann F. and Takagi H., eds., Japan International Research Center for Agricultural Sciences and International Plant Genetic Resources Institute, Rome, Italy, 382-384.  2000.

23.  Chang Y., R. Barker, and B. M. Reed.  Cold acclimation improves recovery of cryopreserved grass (Zoysia and Lolium sp.).  Cryo-Letters 21:107-116.  2000.

  1. Dumet D., Chang Y., Reed B.M. and E.E. Benson.  Replacement of cold acclimatization with high sucrose pretreatment in black currant cryopreservation.  In: Cryopreservation of Tropical Germplasm.  Current research progress and application, Engelmann F. and Takagi H., eds., Japan International Research Center for Agricultural Sciences and International Plant Genetic Resources Institute, Rome, Italy, 385-387.  2000.
  2. Dumet, D., W. Block, R. Worland, B.M. Reed, E.E. Benson.  Profiling cryopreservation protocols for Ribes ciliatum using differential scanning calorimetry.  Cryo-Letters 21:367-378.  2000.
  3. Reed B.M. Genotype consideration in temperate fruit crop cryopreservation.  In: Cryopreservation of Tropical Germplasm.  Current research progress and application, Engelmann F. and Takagi H., eds., Japan International Research Center for Agricultural Sciences and International Plant Genetic Resources Institute, Rome, Italy, 200-204.  2000.
  4. Reed B.M., Brennan R.M. and E.E. Benson.  Cryopreservation:  An in vitro method for conserving Ribes germplasm in international gene banks.  In: Cryopreservation of Tropical Germplasm.  Current research progress and application, Engelmann F. and Takagi H., eds., Japan International Research Center for Agricultural Sciences and International Plant Genetic Resources Institute, Rome, Italy, 470-473.  2000.
  5. Reed B.M., DeNoma J. and Y. Chang.  Application of cryopreservation protocols at a clonal genebank.  In: Cryopreservation of Tropical Germplasm.  Current research progress and application, Engelmann F. and Takagi H., eds., Japan International Research Center for Agricultural Sciences and International Plant Genetic Resources Institute, Rome, Italy, 246-249.  2000.

29.  Chang Y. and B.M. Reed.  Preculture Conditions Influence Cold Hardiness and Regrowth of Pyrus cordata Shoot Tips after Cryopreservation.  HortScience. 36: 1329-1333.  2001.

30.  Reed, B.M.  Implementing Cryogenic Storage of Clonally Propagated Plants.  CryoLetters.  22:97-104. 2001.

31.  Reed, B.M. and K.E. Hummer.  Long-term storage of hazelnut embryonic axes in liquid nitrogen.  Acta Horticulturae.  556: 177-179.  2001

  1. Reed, B.M., D. Dumet, J. M.  DeNoma and E.E. Benson.  Validation of cryopreservation protocols for plant germplasm conservation: A pilot study using Ribes L. Biodiversity and Conservation 10(6): 939-949.  2001.

33.  Reed, B.M., S. Schwanke, and R. Shala.  Pear seeds retain viability after liquid nitrogen immersion.  HortScience. 36: 1121-1122.  2001.

34.  Reed, B.M. Implementing Cryopreservation for Long-Term Germplasm Preservation in Vegetatively Propagated Species, p. 22-36.  In: L.E. Towill and Y.P.S. Bajaj (eds.).  Biotechnology in Agriculture and Forestry: Cryopreservation of Plant Germplasm II, Vol. 50.  Springer-Verlag, Berlin.  2002.

35.  Reed, B.M. and K.E. Hummer. Cryopreservation of Ribes, p. 323-343. In: L.E. Towill and Y.P.S. Bajaj (eds.).  Biotechnology in Agriculture and Forestry: Cryopreservation of Plant Germplasm II, Vol. 50.  Springer-Verlag, Berlin.  2002.