Successful Cryopreservation of Vitis Shoot Tips: Novel Pre-treatment Combinations Applied to Nine Species.

Background: Secure back-up of Vitis genetic resource collections requires cryopreservation methods that give long-term survival of clonal germplasm having diverse genetic backgrounds.

Objective: This work sought to increase survival of Vitis shoot tips exposed to liquid nitrogen using combinations of pretreatments and cryoprotection procedures. The new procedure should give high survival of shoot tips from a wide range of Vitis species.

A high-quality apple genome assembly reveals the association of a retrotransposon and red fruit colour.

A complete and accurate genome sequence provides a fundamental tool for functional genomics and DNA-informed breeding. Here, we assemble a high-quality genome (contig N50 of 6.99 Mb) of the apple anther-derived homozygous line HFTH1, including 22 telomere sequences, using a combination of PacBio single-molecule real-time (SMRT) sequencing, chromosome conformation capture (Hi-C) sequencing, and optical mapping.

Chloroplast heterogeneity and historical admixture within the genus Malus.

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Premise Of The Study: The genus Malus represents a unique and complex evolutionary context in which to study domestication. Several Malus species have provided novel alleles and traits to the cultivars. The extent of admixture among wild Malus species has not been well described, due in part to limited sampling of individuals within a taxon.

Cryopreservation of Populus trichocarpa and Salix dormant buds with recovery by grafting or direct rooting.

Background: Methods are needed for the conservation of clonally maintained trees of Populus and Salix. In this work, Populus trichocarpa and Salix genetic resources were cryopreserved using dormant scions as the source explant.

Objective: We quantified the recovery of cryopreserved materials that originated from diverse field environments by using either direct sprouting or grafting.

Factors affecting stress tolerance in recalcitrant embryonic axes from seeds of four Quercus (Fagaceae) species native to the USA or China.

Background And Aims: Quercus species are often considered 'foundation' components of several temperate and/or subtropical forest ecosystems. However, the populations of some species are declining and there is considerable urgency to develop ex situ conservation strategies. In this study, the storage physiology of seeds within Quercus was explored in order to determine factors that affect survival during cryopreservation and to provide a quantitative assessment of seed recalcitrance to support future studies of this complex trait.

Medium- and long-term storage of the Pycnanthemum (mountain mint) germplasm collection.

The United States of America collection of mountain mint (Pycnanthemum Michx.) is held at the USDA-ARS National Clonal Germplasm Repository (NCGR) in Corvallis, Oregon as seed, potted plants and tissue cultures and a long-term storage collection is preserved at the USDA-ARS National Center for Genetic Resources Preservation (NCGRP) in Fort Collins, Colorado. The clonal collection is comprised of 34 accessions as potted plants that are duplicated with 31 accessions stored as in vitro cultures at 4 degrees C in tissue culture bags for medium-term storage at NCGR and as cryopreserved shoot tips in liquid nitrogen at NCGRP for long-term storage.

Cryopreservation of eucalyptus genetic resources.

The long-term preservation of forest genetic resources is a vital part of preserving our forest crops for future generations. Unfortunately, there are few genebanks dedicated to forest trees and very few methods for long-term preservation of forest genetic resources collections aside from field plantings of a limited number of seed-derived or elite clonal individuals. The use of cryopreservation for the long-term storage of elite germplasm is increasingly being used for the long-term preservation of clonal agronomic crops but for forest trees, such as Eucalyptus, the methodology for cryopreservation of diverse genetic resources collections has not been established.

Development of DArT marker platforms and genetic diversity assessment of the U.S. collection of the new oilseed crop lesquerella and related species.

The advantages of using molecular markers in modern genebanks are well documented. They are commonly used to understand the distribution of genetic diversity in populations and among species which is crucial for efficient management and effective utilization of germplasm collections. We describe the development of two types of DArT molecular marker platforms for the new oilseed crop lesquerella (Physaria spp.

Cryopreservation of citrus shoot tips using micrografting for recovery.

The USDA-ARS National Plant Germplasm System (NPGS) and the University of California Citrus Variety Collection maintain more than 888 unique accessions representing 132 taxa of Citrus, Fortunella, and citrus wild species within field, screenhouse, and greenhouse collections. We have identified a cryopreservation method by which Citrus genetic resources that are not maintained in vitro can be successfully conserved. Shoot tips were excised from actively growing vegetative flushes of protected trees.

Rice domestication: histories and mysteries.

Domesticated rice (Oryza sativa) is one of the world's most important food crops, culturally, nutritionally and economically (Khush 1997). Thus, it is no surprise that there is intense curiosity about its genetic and geographical origins, its response to selection under domestication, and the genetic structure of its wild relative, Oryza rufipogon. Studies of Oryza attempting to answer these questions have accompanied each stage of the development of molecular markers, starting with allozymes and continuing to genome sequencing.

Detailed characterization of mechanical properties and molecular mobility within dry seed glasses: relevance to the physiology of dry biological systems.

Slow movement of molecules in glassy matrices controls the kinetics of chemical and physical reactions in dry seeds. Variation in physiological activity among seeds suggests that there are differences in mobility among seed glasses. Testing this hypothesis is difficult because few tools are available to measure molecular mobility within dry seeds.

Increased efficiency using the encapsulation-dehydration cryopreservation technique for Arabidopsis thaliana.

Arabidopsis thaliana shoot tips were successfully cryopreserved using encapsulation-dehydration cryopreservation methods. Between one and seven shoot tips were encapsulated within 4 mm calcium-alginate beads. Beads were formed in the presence of 2 M glycerol + 0.

Application of functional genomics and proteomics to plant cryopreservation.

Plant cryobiology has primarily emerged from the classical fields of cryobiology and plant stress physiology. Cryopreservation tools are now available to geneticists for germplasm preservation and the field itself is advancing significantly through the use of molecular techniques. Long-term preservation of vegetatively propagated tissues can minimize the risks of long-term maintenance under tissue culture or field conditions.

Increased efficiency using the encapsulation-dehydration cryopreservation technique for Arabidopsis thaliana.

Arabidopsis thaliana shoot tips were successfully cryopreserved using encapsulation-dehydration cryopreservation methods. Between one and seven shoot tips were encapsulated within 4 mm calcium-alginate beads. Beads were formed in the presence of 2 M glycerol + 0.

Implications of the pH and temperature of diluted, cooled boar semen on fresh and frozen-thawed sperm motility characteristics.

Boar semen is typically collected, diluted and cooled for AI use over numerous days, or frozen immediately after shipping to capable laboratories. The storage temperature and pH of the diluted, cooled boar semen could influence the fertility of boar sperm. Therefore, the purpose of this study was to determine the effects of pH and storage temperature on fresh and frozen-thawed boar sperm motility end points.

Cryopreservation of dormant buds from diverse Fraxinus species.

Ash (Fraxinus) is an economically important tree genus in the landscape industry, as well as a key component of North American forests, especially in the North Central United States and adjacent regions in Canada. In recent years, the Emerald Ash Borer beetle (Agrilus planipennis) has significantly threatened the survival of native North American Fraxinus species. A dormant-bud cryopreservation technique has been developed as a method to conserve specific clones of ash.

High viability of dormant Malus buds after 10 years of storage in liquid nitrogen vapour.

Three hundred and sixty two Malus accessions from the Canadian Clonal Genebank of Plant Gene Resources of Canada were cryopreserved as dormant buds at the USDA-ARS National Center for Genetic Resources Preservation in 1996. According to grafting data collected on 165 of these accessions in 1999, 80 percent of the accessions had at least 40 percent viability. A subsample of these accessions was processed for cryopreservation by either adjusting the moisture content of the budwood sections containing dormant buds to 32 or 37 percent moisture (fresh weight basis) or by not drying the budwood sections (46 percent moisture fresh weight basis) prior to cooling.

Hydration of Cuphea seeds containing crystallised triacylglycerols.

Seeds that exhibit intermediate storage behaviour seem to die under conventional -18°C storage conditions. Cuphea wrightii A. Gray, C.

Cryopreservation of Arabidopsis thaliana shoot tips.

Development of a successful shoot tip cryopreservation method for Arabidopsis thaliana L. will enable researchers to use molecular tools to study processes important for successful cryopreservation in this model organism. We demonstrate that Arabidopsis can be successfully cryopreserved using either plant vitrification solution 2 (PVS2) or plant vitrification solution 3 (PVS3) as cryoprotectants prior to rapidly cooling shoot tips in liquid nitrogen (LN).

Massive cellular disruption occurs during early imbibition of Cuphea seeds containing crystallized triacylglycerols.

The transition from anhydrobiotic to hydrated state occurs during early imbibition of seeds and is lethal if lipid reserves in seeds are crystalline. Low temperatures crystallize lipids during seed storage. We examine the nature of cellular damage observed in seeds of Cuphea wrightii and C.

Triacylglycerol phase and 'intermediate' seed storage physiology: a study of Cuphea carthagenensis.

Seeds with 'intermediate' storage physiology store poorly under cold and dry conditions. We tested whether the poor shelf life can be attributed to triacylglycerol phase changes using Cuphea carthagenensis (Jacq.) seeds.

Cryopreservation of apple using non-desiccated sections from winter-collected scions.

Winter vegetative buds of Malus species are cryopreserved at USDA-ARS NCGRP to backup genetic resources maintained by field collections. The method uses desiccation of nodal sections prior to cooling but is time and labor intensive, and can damage materials if excessive. Here we tested cooling sections without prior desiccation to improve the efficiency of handling.

Cryopreservation of Malus germplasm using a winter vegetative bud method: results from 1915 accessions.

Cryopreservation using a winter vegetative bud method is being applied to the Malus collection maintained in the field at the USDA-ARS Plant Genetic Resources Unit, Geneva, New York. Winter hardy materials are sent to the USDA-ARS National Center for Genetic Resources Preservation, Fort Collins, CO, for processing. To date 1915 accessions, representing 30 species and 16 interspecific hybrids, have been tested.

Longevity of cryogenically stored seeds.

Though cryogenic storage is presumed to provide nearly infinite longevity to cells, the actual shelf life achieved under ultra-cold temperatures has not been addressed theoretically or empirically. Here, we report measurable changes in germination of dried seeds stored under liquid nitrogen conditions for >10 years. There was considerable variability in the extent of deterioration among species and accessions within a species.