Simultaneous encapsulation of seed and mycorrhizal fungi for long-term storage and propagation of terrestrial orchids

2008 ◽  
Vol 56 (7) ◽  
pp. 609 ◽  
Author(s):  
Karen D. Sommerville ◽  
John P. Siemon ◽  
Chris B. Wood ◽  
Catherine A. Offord
Keyword(s):  
Mycorrhizal Fungi ◽  
Ex Situ Conservation ◽  
Alginate Beads ◽  
Ex Situ ◽  
Storage Duration ◽  
Long Term Storage ◽  
Mycorrhizal Associations ◽  
Terrestrial Orchids

Ex situ conservation of threatened terrestrial orchids requires the simultaneous conservation of their mycorrhizal associations. A method for encapsulating both seed and fungi in alginate beads (known as encapsulation–dehydration) was applied to the storage and propagation of two endangered orchid species in NSW, Australia—Pterostylis saxicola D.L.Jones & M.A.Clem. and Diuris arenaria D.L.Jones. We tested the effect of storage duration and temperature on fungal recovery and germination potential in vitro, and recorded survival for seedlings subsequently transferred to potting mix. Storage at 23°C significantly reduced fungal recovery and germination for both species after only 3 months (P < 0.05), whereas storage at 4°C significantly reduced fungal recovery for P. saxicola after 6 months (P < 0.05). Storage for 6 months at −18 and −196°C had no significant effect on the fungal recovery and germination percentages of either species. All beads transferred directly from in vitro culture to potting mix resulted in the establishment of at least one seedling, and production of a healthy tuberoid, when transferred near the commencement of the natural growing season. The encapsulation–dehydration method may have a practical application for use in ex situ conservation of other terrestrial orchids, as well as their mycorrhizal fungi.

scholarly journals Ex situ conservation of plant species of the Red Book of Russia in botanical gardens

2021 ◽  
Vol 2 (44) ◽  
pp. 11-11
Author(s):  
Alexander Saakian ◽  
◽  
Keyword(s):  
Plant Species ◽  
Russian Federation ◽  
Ex Situ Conservation ◽  
Ex Situ ◽  
Endangered Plants ◽  
Botanical Gardens ◽  
Rare And Endangered ◽  
The Russian Federation

Russian botanical gardens are actively involved in the conservation of rare and endangered plants. They pay special attention to the species included in the Red Book of the Russian Federation. At present, 377 species of higher plants are cultivated in living collections of botanical gardens out of 514 species presented in the Red Book of Russia, which is 73%. Thus, the Russian Federation has practically met the requirement of goal 8 of the Global Strategy for Plant Conservation. The vast majority of rare plant species are represented by samples in the collections of many botanical gardens and the reliability of their protection under cultural conditions is beyond doubt. In the Main Botanical Garden of the Russian Academy of Sciences, an in vitro collection of rare and endangered plants is preserved, including 82 species, which is 17.3% of the total number of angiosperms included in the Red Book of the Russian Federation. As a result of many years of research, the features of the cultivation and preservation of ex situ plants belonging to different families have been revealed. The main methodological aspects at the stages of obtaining a sterile culture, micropropagation proper and long-term deposition are reflected. The compositions of nutrient media and cultivation factors were optimized for slowed growth of explants of the studied cultures and preservation of their viability. Keywords: RARE AND ENDANGERED PLANT SPECIES, RED BOOK OF THE RUSSIAN FEDERATION, EX SITU CONSERVATION, LONG-TERM IN VITRO CONSERVATION

scholarly journals Propagation and reintroduction of Caladenia

2009 ◽  
Vol 57 (4) ◽  
pp. 373 ◽  
Author(s):  
Magali Wright ◽  
Rob Cross ◽  
Kingsley Dixon ◽  
Tien Huynh ◽  
Ann Lawrie ◽  
...  
Keyword(s):  
Conservation Planning ◽  
Mycorrhizal Fungi ◽  
Ex Situ ◽  
Terrestrial Orchid ◽  
Natural Habitats ◽  
Nurse Plants ◽  
Mycorrhizal Associations ◽  
Symbiotic Germination ◽  
Growing Conditions

Many Caladenia species have been reduced to extremely small and/or fragmented populations, and reintroduction/translocation into natural or rehabilitated habitats, by using ex situ propagated plants or via direct seeding, represents an important adjunct in conservation planning. However, Caladenia species are some of the most difficult terrestrial orchid taxa to propagate, in part because of the specificity of the mycorrhizal associations and the need to provide growing conditions that suit both the mycorrhizal fungi and Caladenia plants. The present paper reviews recent advances in Caladenia propagation and reintroduction methods, including in vitro seed germination, transferral from in vitro to nursery environments, ex vitro symbiotic germination (germination in inoculated nursery media), nursery cultivation, the use of nurse plants and reintroduction of Caladenia into natural habitats by using seed, dormant tubers or growing plants. Techniques discussed in the present paper increase the options for future Caladenia conservation programs, especially for those species currently on the brink of extinction.

Codiversification of orchids (Pterostylidinae) and their associated mycorrhizal fungi

2011 ◽  
Vol 59 (5) ◽  
pp. 480 ◽  
Author(s):  
J. Tupac Otero ◽  
Peter H. Thrall ◽  
Mark Clements ◽  
Jeremy J. Burdon ◽  
Joseph T. Miller
Keyword(s):  
Mycorrhizal Fungi ◽  
Mycorrhizal Fungus ◽  
Distinct Group ◽  
Host Group ◽  
Mycorrhizal Associations ◽  
As Species ◽  
Terrestrial Orchids ◽  
Nuclear Its ◽  
Germination Experiments

Fungal symbionts involved in mycorrhizal associations are known to vary considerably in both specificity and the level of benefits conferred on their plant hosts. For orchids, association with a suitable mycorrhizal fungus is vital for successful germination, growth and establishment. Using an evolutionarily distinct group of Australasian terrestrial orchids, the Pterostylidinae (Cranichiadeae: Orchidaceae), we assessed potential codiversification and the level of response between this diverse host group (~250 species) and their associated fungal symbionts. All fungal isolates recovered (~200 from 41 host species covering all major orchid clades) were identified as species of Ceratobasidium, which clustered into strongly supported groups using nuclear (ITS) and mitochondrial (ML 4–5) gene sequences. Three clades within the Pterostylidinae phylogeny showed associations with specific fungal clades. The results suggest the occurrence of local adaptation by the fungal symbionts to the orchid host, particularly in diverse and widespread host taxa. Results of cross-inoculation in vitro germination experiments revealed correlations between certain mycorrhizal fungal clades and particular orchid taxa, with germination generally being most effective when seeds were inoculated with fungal strains from the same clade as found naturally associated with the orchid species. We found only general congruence between the orchid and fungal phylogenies, suggesting that strict codivergerence between these orchids and their mycorrhizal associates has not occurred at the broad level of resolution studied.

scholarly journals Media Composition and Light Affect Storability and Poststorage Recovery of Micropropagated Hosta Plantlets

HortScience ◽  
2000 ◽  
Vol 35 (6) ◽  
pp. 1159-1162 ◽  
Author(s):  
Sandra B. Wilson ◽  
Nihal C. Rajapakse ◽  
Roy E. Young
Keyword(s):  
Storage Temperature ◽  
Visual Quality ◽  
Dry Weight ◽  
Poor Quality ◽  
Photon Flux ◽  
Storage Duration ◽  
Long Term Storage ◽  
Leaf Yellowing

Hosta (Hosta tokudama Makeawa `Newberry Gold') plantlets were micropropagated photoautotrophically (without sucrose in medium) or photomixotrophically (with 2% sucrose in medium) for 3 weeks at 23 °C under 80 μmol·m-2·s-1 photosynthetic photon flux (PPF) prior to long-term storage. Plantlets were stored for 4, 8, or 12 weeks at 5, 10, or 22 °C in darkness or under white (400-800 nm), blue (400-500 nm), or red (600-700 nm) light at or near light compensation points. Illumination during storage was necessary to maintain dry weight and regrowth potentials of plantlets in vitro, but light quality had no effect on these parameters. All photoautotrophic plantlets stored in darkness were of poor quality at the time of removal from storage and died when transferred to the greenhouse. Dark-stored photomixotrophic plantlets survived storage for 12 weeks at 5 °C, but declined in appearance (visual quality) as the storage duration increased. Decline in visual quality was greater when plantlets were stored at 10 and 22 °C. Leaf dry weight of illuminated plantlets increased and percentage of leaf yellowing decreased as storage temperature increased. Recovery of illuminated plantlets from photomixotrophic storage was best when plantlets were stored at 22 °C. These plantlets were characterized by increased visual quality (color and form) and increased dry weight compared with those in other treatments. After 60 days in the greenhouse, the dry weight of these plantlets was similar for 4-, 8-, and 12-week storage durations, indicating flexibility in storage time if specific light and temperature provisions are met.

scholarly journals Kriopreservasi untuk Konservasi Plasma Nutfah Tanaman: Peluang Pemanfaatannya di Indonesia

2016 ◽  
Vol 3 (2) ◽  
pp. 80
Author(s):  
Semuel Leunufna
Keyword(s):  
Ex Situ Conservation ◽  
Germplasm Collection ◽  
Ex Situ ◽  
Seed Plants ◽  
High Recovery ◽  
High Recovery Rate ◽  
Long Term Preservation ◽  
Collection Methods

<p>Increasing rate of plant germplasm lost in<br />Indonesia has promoted the implementation of various<br />methods for their conservation. Cryopreservation is a technique<br />applicable for a long-term preservation (base collection)<br />of plants possessing non-orthodox (recalcitrant and<br />semi-recalcitrant) seeds and those propagated vegetatively.<br />The technique can be used as an alternative method for<br />orthodox seed plants preservation in the ex situ conservation<br />system. Although field and in vitro collection methods can<br />be applied for the non-orthodox seed plants, a number of<br />disadvantages possesed by these methods, especially in the<br />tropics or the developing countries, deny their use for the<br />establishment of a long-term germplasm collection. Successful<br />implementation of the cryopreservation technique is<br />supported by the development of protocols, which are able<br />to provide a high recovery rate for species understudy, using<br />vitrification based methods which are simple, economical,<br />applicable to complex organs, and able to implement a high<br />number of explants per experiment. The availability of infrastructures<br />including in vitro culture laboratories, continue<br />supply of liquid nitrogen is highly supporting the use of<br />cryopreservation technique in Indonesia.</p>

Long-term storage of mycorrhizal fungi and seed as a tool for the conservation of endangered Western Australian terrestrial orchids

2001 ◽  
Vol 49 (5) ◽  
pp. 619 ◽  
Author(s):  
A. L. Batty ◽  
K. W. Dixon ◽  
M. Brundrett ◽  
K. Sivasithamparam
Keyword(s):  
Liquid Nitrogen ◽  
Silica Gel ◽  
Mycorrhizal Fungi ◽  
Seed Viability ◽  
Seed Storage ◽  
Ex Situ ◽  
Testing Procedures ◽  
Terrestrial Orchids ◽  
The Impact

The impact of seed drying, seed storage and development of testing procedures for seed viability assessment was undertaken for a selection of common taxa with congeners that are rare and endangered (Caladenia, Diuris, Pterostylisand Thelymitra). Freshly collected seed showed significantly lower levels of germination compared with seed that had been subjected to drying over silica gel for 24 h. Seed dried over silica gel for 24 h and plunged into liquid nitrogen exhibited a further increase in germination levels. Germination of seed stored at 4, 18 or 22˚C for 1 year was substantially higher than freshly collected seed (4 weeks after dehiscence), but germination was highest overall after storage of dried seed in liquid nitrogen (–196˚C). Mycorrhizal fungi that promote the germination and growth of plants were also successfully preserved in liquid nitrogen. The use of cryoprotectants on fungal isolates had no observable deleterious effects on fungal regeneration. Histochemical staining procedures (tetrazolium, fluorescein diacetate and Evans blue) substantially overestimated seed viability, relative to symbiotic seed germination, for most seed treatments indicating a need for re-evaluation of the effectiveness of staining procedures for testing viability. The implications of the long-term ex situ storage of orchid seed and fungal symbionts for the conservation of endangered orchids is discussed.

scholarly journals In vitro propagation of endangered Dianthus taxa

2015 ◽  
pp. 83-98
Author(s):  
Marija Markovic ◽  
Mihailo Grbic ◽  
Matilda Djukic
Keyword(s):  
In Vitro Culture ◽  
In Vitro Propagation ◽  
Ex Situ Conservation ◽  
Ex Situ ◽  
Medium Term ◽  
Research And Practice

The review of recent researches regarding the in vitro culture of 30 endangered Dianthus taxa is presented in this paper. Various in vitro protocols developed for selected rare and threatened Dianthus taxa are analysed in order to provide a useful synthesis of the data obtained with the main principles, techniques and recommendations for futher research and practice. The recapitulated data presented in this review can be used as a tool for the micropropagation of other endangered Dianthus taxa, enabling their propagation and obtaining a sufficient amount of plants for reintroduction. In addition, the obtained results represent the basis for ex situ conservation of the investigated taxa, especially for medium-term and long-term conservation (cryopreservation).

Endogone pisiformis : croissance, morphologie et conservation des cultures

1990 ◽  
Vol 68 (4) ◽  
pp. 910-915 ◽  
Author(s):  
Yolande Dalpé
Keyword(s):  
Mycorrhizal Fungi ◽  
Pure Culture ◽  
Culture Media ◽  
Arbuscular Mycorrhizal ◽  
Culture Collection ◽  
Long Term Storage ◽  
Vesicular Arbuscular

The species Endogone pisiformis can now be maintained in pure culture collection in its vegetative form. In vitro, morphological observations of the mycelium revealed the presence of ramified, coenocytic hyphae, which bore both thin- and thick-walled swellings analogous to the vesicles and chlamydospores of vesicular-arbuscular mycorrhizal fungi of the genera Glomus and Sclerocystis. Similar mycelial structures also have been observed, in vivo, on Sphagnum plants along vegetative hyphae still attached to sporocarps. Natural, semisynthetic, and synthetic culture media supported the growth of E. pisiformis for a few successive transfers, but the strains then rapidly lost their vitality. With the addition of thiamine to the culture media, strains completely recovered their vitality. Using thiamine, long-term storage by lyophilisation or under mineral oil is possible. Key words: Endogone pisiformis, fungal morphology, fungal pure culture, thiamine.

scholarly journals Efficient Cryopreservation of Populus tremula by In Vitro-Grown Axillary Buds and Genetic Stability of Recovered Plants

Plants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 77
Author(s):  
Elena O. Vidyagina ◽  
Nikolay N. Kharchenko ◽  
Konstantin A. Shestibratov
Keyword(s):  
Survival Rate ◽  
Axillary Buds ◽  
Long Term Storage ◽  
Average Survival ◽  
Transgenic Lines ◽  
Ssr Loci ◽  
One Step ◽  
The One

Axillary buds of in vitro microshoots were successfully frozen at –196 °C by the one-step freezing method using the protective vitrification solution 2 (PVS2). Microshoots were taken from 11 transgenic lines and three wild type lines. Influence of different explant pretreatments were analyzed from the point of their influence towards recovery after cryopreservation. It was found out that the use of axillary buds as explants after removal of the apical one increases recovery on average by 8%. The cultivation on growth medium of higher density insignificantly raises the regenerants survival rate. Pretreatment of the osmotic fluid (OF) shows the greatest influence on the survival rate. It leads to the increase in survival rate by 20%. The cryopreservation technology providing regenerants average survival rate of 83% was developed. It was based on the experimental results obtained with explant pretreatment. Incubation time in liquid nitrogen did not affect the explants survival rate after thawing. After six months cryostorage of samples their genetic variability was analyzed. Six variable simple sequence repeat (SSR) loci were used to analyze genotype variability after the freezing-thawing procedure. The microsatellite analysis showed the genetic status identity of plants after cryopreservation and of the original genotypes. The presence of the recombinant gene in the transgenic lines after cryostorage were confirmed so as the interclonal variation in the growth rate under greenhouse conditions. The developed technique is recommended for long-term storage of various breeding and genetically modified lines of aspen plants, as it provides a high percentage of explants survival with no changes in genotype.

scholarly journals Cryopreservation of Woody Crops: The Avocado Case

Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 934
Author(s):  
Chris O’Brien ◽  
Jayeni Hiti-Bandaralage ◽  
Raquel Folgado ◽  
Alice Hayward ◽  
Sean Lahmeyer ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Ex Situ Conservation ◽  
Plant Genetic Resources ◽  
Ex Situ ◽  
Conservation Strategies ◽  
Recalcitrant Seed ◽  
In Vitro Storage ◽  
Germplasm Collections ◽  
Seed Crops

Recent development and implementation of crop cryopreservation protocols has increased the capacity to maintain recalcitrant seeded germplasm collections via cryopreserved in vitro material. To preserve the greatest possible plant genetic resources globally for future food security and breeding programs, it is essential to integrate in situ and ex situ conservation methods into a cohesive conservation plan. In vitro storage using tissue culture and cryopreservation techniques offers promising complementary tools that can be used to promote this approach. These techniques can be employed for crops difficult or impossible to maintain in seed banks for long-term conservation. This includes woody perennial plants, recalcitrant seed crops or crops with no seeds at all and vegetatively or clonally propagated crops where seeds are not true-to-type. Many of the world’s most important crops for food, nutrition and livelihoods, are vegetatively propagated or have recalcitrant seeds. This review will look at ex situ conservation, namely field repositories and in vitro storage for some of these economically important crops, focusing on conservation strategies for avocado. To date, cultivar-specific multiplication protocols have been established for maintaining multiple avocado cultivars in tissue culture. Cryopreservation of avocado somatic embryos and somatic embryogenesis have been successful. In addition, a shoot-tip cryopreservation protocol has been developed for cryo-storage and regeneration of true-to-type clonal avocado plants.

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