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United States Department of Agriculture

Agricultural Research Service



2011 Annual Report

1a. Objectives (from AD-416)
Objective 1: Develop physiological and biophysical approaches and tools to assess changes in plant germplasm viability and the potential causes during genebank preservation. Objective 2: Develop statistical genetic strategies and tools to sample and preserve plant genetic diversity in genebank collections and in situ reserves.

1b. Approach (from AD-416)
The Preservation of Plant Genetic Diversity in Ex Situ Genebank program scientists conduct research to improve the biological and genetic integrity of genebanked germplasm and to standardize procedures for handling accessions and reporting associated data. Interrelated research goals will allow curators to preserve viability of conservation targets (Objective 1) and rationalize and validate the genetic diversity and integrity of those targets (Objective 2). Using taxa that are empirically tractable systems, we will • define tolerances to preservation stresses of selected propagules, • develop methods to improve survival or reliably predict loss of viability over time, • model the effects of mortality and regeneration on genetic composition, and • develop sampling strategies for wild-collected germplasm that maximize genetic diversity while minimizing curator inputs for storage and regeneration. A central theme is identifying appropriate conservation targets that capture desired genetic diversity, remain viable during storage and are available to the user when needed. A conservation target is a group of propagules (such as seeds or pollens) or an individual propagule (such as an explant) that comprises an accession valued for specific genes, genetic richness (number and frequency of alleles) or an allelic combination (genotype). PGPRU scientists and their collaborators will investigate major conceptual issues of repository biology and standardization using within-unit expertise in biophysics, plant physiology, cell and molecular biology and population genetics and National Plant Germplasm System (NPGS) curators’ expertise on reproductive biology, phenotypic diversity, history and cultivation of their assigned collections. Our central position within NPGS allows us to develop protocols and predictive tools that are applicable to a wide variety of species and propagules.

3. Progress Report
Interrelated research allows curators to preserve viability of conservation targets and rationalize and validate the genetic diversity and integrity of those targets. Survival of germplasm exposed to cryostorage conditions becomes increasingly possible through research on mechanisms of damage and protection of cells, tissues and organs. Genotypes vary significantly in response to stress, protection and recovery media and this variation requires considerable effort to optimize preservation procedures for individual accessions. Understanding the basis for different responses among plant cells will lead to greater efficiency in cryopreservation efforts. Major advances in developing recovery conditions for diverse citrus and avocado shoot tips has helped to streamline assessments of cryopreservation procedures. Efforts continue to describe the early stage of seed aging that is currently regarded as “asymptomatic.” Kinetics of aging and other reactions that occur under dry conditions, such as dormancy breaking and color changes, are correlated with biophysical measurements related to molecular mobility and phase behavior and biochemical changes such as emission of volatiles and degradation products of lipids, proteins and nucleic acids. Moisture and temperature effects of these reactions are used to predict the duration of the asymptomatic phase. Aging of preserved inventories is inevitable and older inventories must be regenerated. Both the aging and regeneration process can lead to loss of genetic identity of inventories. Population genetic tools are an essential component of NCGRP research to link efficiency and accountability to preservation methods. The extent and mechanisms of genetic change during genebanking are studied by changes in quantitative traits and genomic analysis of neutral markers and expressed sequences. Consequences of seed aging and regeneration have been modeled, and these models are now being tested with inventory grow outs. The expected product is an assessment of the degree to which random changes and directed selection affect genetic integrity of genebanked samples and the time period in which these changes are expected to occur. This assessment of the effectiveness of genebanking is critical to accommodate the increased attention of wild-collected accessions. As collections become larger and larger, it becomes increasingly important to develop tools to express, quantify, distill and communicate the genetic diversity of NPGS collections. These tools necessarily require integration of genetic and geospatial data. Advanced methods in statistical genetic structure estimation and GIS mapping play a critical role in visualizing patterns of diversity and help locate potential gaps in collections.

4. Accomplishments

Review Publications
Reeves, P.A., Richards, C.M. 2011. Species delimitation under the general lineage concept: An empirical example using wild North American hops (cannabaceae: Humulus lupulus). Systematic Biology. 60:45-59.

Gross, B.L. 2011. MADS-box out of the black box. Molecular Ecology. 20:25-26.

Richards, C.M., Lockwood, D.R., Volk, G.M., Walters, C.T. 2010. Modeling demographics and genetics in ex situ collections during seed storage and regeneration. Crop Science. 50:2440-2447.

Volk, G.M. 2010. Advantages for the use of standardized phenotyping in databases. HortScience 45:1310-1313.

Bassil, N.V., Volk, G.M. 2010. Standardized Phenotyping: Advantages to Horticulture, Introduction to the Workshop. HortScience. 45(9):1306.

Gross, B.L., Strasburg, J.L. 2010. Cotton domestication: Dramatic changes in a single cell. BioMed Central Biology. 8:137-139.

Njuguna, W., Hummer, K.E., Richards, C.M., Davis, T.M., Bassil, N.V. 2011. Genetic diversity of diploid Japanese strawberry species based on microsatellite markers. Genetic Resources and Crop Evolution. 58:1187-1198.

Mira, S., Gonzalez-Benito, E., Hill, L.M., Walters, C.T. 2010. Characterization of volatile production during storage of lettuce (Lactuca sativa) seed. Journal of Experimental Botany. 61:3915-3924.

Walters, C.T., Ballesteros, D., Vertucci, V. 2011. Structural mechanics of seed deterioration: Standing the test of time. Plant Science. 179:565-573.

Ballesteros, D., Estrelles, E., Walters, C.T., Ibars, A.M. 2011. Effect of temperature on green spore longevity for the ferns Equisetum ramosissimum and Osmunda regalis. CryoLetters. 32:89-98.

Thorp, K.R., Dierig, D.A., French, A.N., Hunsaker, D.J. 2010. Analysis of hyperspectral reflectance data for monitoring growth and development of lesquerella. Industrial Crops and Products. 33(2):524-531.

Postman, J.D., Volk, G.M., Aldwinckle, H. 2010. Standardized plant disease evaluations will enhance resistance gene discovery. HortScience. 45(9):1317-1320.

Last Modified: 10/19/2017
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