The contribution of in vitro technology and cryogenic storage to conservation of indigenous plants

2007 ◽  
Vol 55 (3) ◽  
pp. 345 ◽  
Author(s):  
Eric Bunn ◽  
Shane Turner ◽  
Maggie Panaia ◽  
Kingsley W. Dixon
Keyword(s):  
Tissue Culture ◽  
Mycorrhizal Fungi ◽  
Ex Situ ◽  
Culture Methods ◽  
Germplasm Storage ◽  
Indigenous Plants ◽  
Cryogenic Storage ◽  
Propagation Methods ◽  
Research Initiatives

In vitro culture has enabled a variety of recalcitrant and threatened plant taxa to be micropropagated in the absence of viable conventional propagation methods. Cryogenic storage research has provided alternative protocols for efficient long-term germplasm storage for many plant species. Recent advances in tissue-culture methods such as somatic embryogenesis have enabled the production of >20 000 somatic embryos of a recalcitrant native Australian rush in a few months, far higher than other in vitro methods for these types of plants. Cryogenic protocols are reported for >30 species of Australian vascular plants, seed and numerous mycorrhizal fungi (mainly orchid spp.), greatly extending the range and type of material that can be stored through the application of cryogenic methods. The role of in vitro and cryogenic research initiatives in botanic gardens for plant biodiversity conservation and restoration is discussed, using examples of successful ex situ conservation through tissue-culture and cryogenic-storage research.

scholarly journals Application of Horticultural and Tissue Culture Methods for Ex Situ Conservation of Endangered Primula farinosa L.

2020 ◽  
Vol 89 (1) ◽  
Author(s):  
Ewa Sitek ◽  
Barbara Nowak ◽  
Michał Fecowicz ◽  
Zbigniew Gajewski ◽  
Piotr Dańda ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Culture Conditions ◽  
Ex Situ ◽  
In Vitro Cultivation ◽  
Culture Methods ◽  
Transient Nature ◽  
Micropropagated Plants ◽  
The Difference

Our study aimed at active conservation of the last location of <em>Primula farinosa</em>, an endangered species in Poland, and assessed reproduction by seeds and plant propagation on sterile media in tissue culture conditions. We identified gibberellic acid (GA<sub data-id="subscript-1">3</sub>) as the key factor stimulating germination of <em>P. farinosa</em> seeds. Growing juvenile plants under controlled temperature of 18/16 °C day/night yielded good quality plant material without mycorrhization. In tissue culture, the most favorable medium for shoot propagation was MS supplemented with the lowest tested concentration of indole-3-butyric acid (IBA; 0.05 mg dm<sup data-id="superscript-1">−3</sup>) and 6-benzyl-aminopurine (BAP; 0.1 mg dm<sup data-id="superscript-2">−3</sup>). The rooting ability of shoots was high and comparable for all auxins used. 2C DNA content of seed-derived and micropropagated plants did not indicate any change in the ploidy level during in vitro cultivation. Plants derived from seeds and tissue cultures were compared in a 2-year study. Of all the characteristics compared, only the number of flowers per inflorescence was lower for micropropagated plants when compared with the seed-origin plants in the first year of observation. The difference was of transient nature and was not observed in the second year of the study. Effective protocols for in vivo and in vitro propagation of <em>P. farinosa</em> were developed, which can be used in practical species protection.

scholarly journals Micropropagation of the Narrow Endemic Hladnikia pastinacifolia (Apiaceae)

2016 ◽  
Vol 75 (2) ◽  
pp. 244-252 ◽  
Author(s):  
Jana Ambrožič-Dolinšek ◽  
Terezija Ciringer ◽  
Mitja Kaligarič
Keyword(s):  
Tissue Culture ◽  
Growth Regulators ◽  
Ex Situ ◽  
Distribution Area ◽  
Ms Medium ◽  
Axillary Shoots ◽  
Culture Techniques ◽  
Narrow Endemic ◽  
Propagation Methods

Abstract The monotypic Hladnikia pastinacifolia Rchb. is a narrow endemic species, with an extremely small distribution area in Slovenia, prone to any kind of threat that could lead to species extinction. Tissue culture techniques are proposed as a conservation measure for rapid propagation and ex-situ conservation. Tissue culture was initiated from seeds and juvenile plants obtained from natural sites on a solid Murashige and Skoog (MS) medium, with and without growth regulators. We tested various combinations and concentrations of growth regulators, and the best proliferation of axillary shoots, on average 14, was obtained on MS medium with 5 μM BAP and 3 μM IBA and 3% sucrose. Rooting was achieved after transferral of the shoots to an MS medium with 2 μM IBA and 3% sucrose. The rooted plants were acclimatized on a mixture of limestone sand, potting soil and vermiculite in a ratio of 10:2:2, with pH in the range of 7.5–8.0. In vitro propagation methods provide an important opportunity for the propagation and preservation of H. pastinacifolia by rapidly increasing the number of plants, without disturbing the wild population.

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.

scholarly journals Artificial Cultivation System for Gastrodia spp. and Identification of Associated Mycorrhizal Fungi

2017 ◽  
Vol 9 (4) ◽  
pp. 27 ◽  
Author(s):  
Chie Shimaoka ◽  
Hirokazu Fukunaga ◽  
Seishu Inagaki ◽  
Shinichiro Sawa
Keyword(s):  
Mycorrhizal Fungi ◽  
Flowering Plants ◽  
Ex Situ ◽  
Natural Conditions ◽  
Cultivation System ◽  
Horticultural Plants ◽  
The Relationship ◽  
Single Tuber ◽  
Artificial Cultivation

The Orchidaceae are the largest and most diverse family of flowering plants on earth, and include some of the most important horticultural plants. While mycoheterotrophic orchids belonging to the genus Gastrodia are known to be provided with carbon through mycorrhizal fungi, the relationship between the plants and fungi is poorly understood. Furthermore, it is challenging to cultivate Gastrodia spp. in vitro. In this study, we present an efficient method for germinating Gastrodia pubilabiata (Gp), Gastrodia nipponica (Gn), and Gastrodia confusa (Gc) plants in vitro, which results in the production of a protocorm and tuber, as under natural conditions. The Gp and Gc plants produced flowers 126 and 124 days after germination, respectively, and set seed under our artificial conditions. In addition, Gp plants flowered up to three times a year from a single tuber. Using our artificial cultivation system, we identified some of the mycorrhizal fungi associated with these plants. Gastrodia spp. appear to obtain carbon from many kinds of mycorrhizal fungi. Our artificial cultivation method is a rapid and efficient means of growing Gastrodia spp. In addition to having applications in research and commercial nurseries, this method could be used to conserve Gastrodia spp. in ex situ, many of which are endangered.

Tests by Tissue Culture Methods on the Nature of Immunity Transplanted Skin

1948 ◽  
Vol s3-89 (7) ◽  
pp. 239-252
Author(s):  
P. B. MEDAWAR
Keyword(s):  
Tissue Culture ◽  
Cell Division ◽  
Epidermal Cell ◽  
Cell Suspensions ◽  
Acquired Immunity ◽  
Culture Methods ◽  
Division Frequency

The transplantation of skin from one rabbit to another elicits a reaction that conforms in main outline with that of an actively acquired immunity. The experiments described in this paper were designed to test the hypothesis that the regression of such grafts is secured by the action of antibodies demonstrable in vitro. Skin from adult rabbits has therefore been cultivated in the presence of serum and growing mesenchymal tissues derived solely from rabbits heavily and specifically immunized against it. Immune sera and tissues are without effect on the survival, cell-division frequency and migratory activities of explanted skin, and agglutinins for epidermal cell suspensions are not demonstrable in immune sera. With certain stated qualifications, it has therefore been concluded that the occurrence of free antibodies is not a sufficient explanation of the regression of skin homografts in vivo.

scholarly journals 83ANALYSIS OF RAPID COOLING V. SLOW COOLING COMBINED WITH ICE CRYSTAL SEEDING FOR CRYOPRESERVATION OF PRIMATE EMBRYONIC STEM CELLS

2004 ◽  
Vol 16 (2) ◽  
pp. 163
Author(s):  
S. Baran ◽  
C. Ware
Keyword(s):  
Tissue Culture ◽  
Es Cells ◽  
Embryonic Stem ◽  
Ice Crystal ◽  
Culture Methods ◽  
Slow Cooling ◽  
Rapid Cooling ◽  
Alkaline Phosphatase Staining

Primate embryonic stem (ES) cells have the ability to self-renew indefinitely while maintaining the ability to differentiate. This unique property allows scientists to study the factors necessary for stem cell self-renewal and differentiation in vitro that reflect in vivo processes. Work with primate ES cells is handicapped by the poor survival (1–5%) of rhesus and human ES cells following standard tissue culture methods of rapid cryopreservation. The purpose of this study was to compare and contrast two cryopreservation techniques, slow cooling combined with ice crystal seeding commonly used for mammalian embryos v. rapid cooling commonly used for tissue culture, to find a method for efficient primate ES cell cryopreservation. A combination of trials was run to compare dimethyl sulfoxide (DMSO) v. ethylene glycol as a cryoprotectant, a cooling rate of 0.3°C per minute following ice crystal seeding at −7°C v. placement at −80°C with no seeding, and rapid thaw with step-wise cryoprotectant removal v. one-step sucrose cryoprotectant removal. Cell survival was assessed through a combination of cell surface markers, alkaline phosphatase staining and morphology to look for undifferentiated cells and quantitate survival. All cryopreservations were performed with the same cell density. The survival of the cells with slow embryo-style cooling in DMSO with a step-wise cryoprotectant removal was 64.0% v. 12.8% with rapid cooling.

scholarly journals Study of Seaweed Kappaphycus alvarezii Explants Growth in the Different Salinity Concentrations

2021 ◽  
Vol 13 (1) ◽  
pp. 97
Author(s):  
Muhammad Aris ◽  
Fatma Muchdar ◽  
Rusmawati Labenua
Keyword(s):  
Tissue Culture ◽  
Kappaphycus Alvarezii ◽  
Environmental Parameters ◽  
Culture Method ◽  
Salinity Treatment ◽  
Culture Methods ◽  
Culture Techniques ◽  
Randomized Design ◽  
Salinity Levels

HighlightThe best salinity for the thallus growth of K. alvarezii is 32 mg/LThe salinity indicates the osmotic balance of K. alvarezii seaweedThis osmoregulation process affects the nutrient absorption of K. alvarezii seaweedThis study aims to develop the availibility of superior seeds of K. alvareziiAbstract Kappaphycus alvarezii is one of the leading commodities in Indonesian waters. Demand for this commodity is quite high, as reflected in the increasing volume of exports each year. Fulfillment of these demands is obtained from the production of cultivation. Generally farmers get natural seaweed seedlings, namely cuttings from existing seaweed. The continuous use of seeds from nature can cause deterioration in the quality and quantity. Handling the problem of quality deterioration from seaweed seeds originating from nature, can be overcome by multiplying the seeds through tissue culture methods in vitro. In term of tissue culture techniques method, the most important thing to note is environmental parameters. An environment parameter that changes suddenly such as salinity can inhibit the growth of seaweed. Thus, this work is attampting the different salinity treatment on seaweed explants K. alvarezii. This study aims at determining (weight) the explants of K. alvarezii with a comparison of the different salinity levels in the in vitro tissue culture method. The method used in this study was a completely randomized design (CRD) with the different salinity treatments namely 30, 31, 32, 33, and 34 ppt. The results showed that the different salinities influenced the growth rate of K. alvarezii seaweed explants with the best explant growth at the salinity of 31 ppt, while the lowest growth value was obtained at 34 ppt

scholarly journals Dünyada ve Türkiye’de Zeytinde Yapılan Doku Kültürü Çalışmaları

2020 ◽  
Vol 8 (3) ◽  
pp. 645
Author(s):  
Zeliha Çiftçi ◽  
Mizgin Ay ◽  
Ebru Sakar
Keyword(s):  
Tissue Culture ◽  
Culture Methods ◽  
Important Place ◽  
Fruit Species ◽  
Mediterranean Countries ◽  
Micro Propagation ◽  
History Of ◽  
The Mediterranean ◽  
Olive Varieties

Known as the world’s most healthy and natural source of vegetable oil, the history of olives dates back to 10,000 years ago. The homeland of olives, a member of the Oleacea family, is Upper Mesopotamia and Southern Asia, including Southeastern Anatolia and Syria. Olives, BC It started to be cultivated on the eastern shores of the Mediterranean in the year 3000 and is one of the first fruit species cultivated in the Mediterranean region. In this respect, olive has an important place in the economy, nutrition and culture of Mediterranean countries. Currently, in most olive growing countries, olive, leafy stem or cuttings are rooted or by propagating stem shoots from seed or clonal stem. However, the so-called table olives are very difficult or completely impossible to root. The olives, which are very difficult to root, should be supported with biotechnological approaches such as micropropagation method in order to increase the product productivity. So far, many fruit species have been propagated in vitro using tissue culture methods and at the same time, some olive varieties have been successfully propagated by micro-propagation method. It made in tissue culture in the world and Turkey Olives have been compiled resources to work for the researchers in this study.

The use of in vitro methods for plant genetic conservation

1982 ◽  
Vol 11 (2) ◽  
pp. 67-72 ◽  
Author(s):  
C. P. Wilkins ◽  
T. Bengochea ◽  
J. H. Dodds
Keyword(s):  
Tissue Culture ◽  
Genetic Material ◽  
Genetic Conservation ◽  
Fundamental Importance ◽  
Culture Methods ◽  
In Vitro Methods ◽  
Controlled Conditions

The preservation of genetically stable tissue for future propagation is of fundamental importance to plant breeders. In many cases this can be done by storing seed under carefully controlled conditions but there are many plants for which this is not possible or may not be economically feasible. This article reviews current techniques of long-term conservation of plant genetic material by tissue culture methods.

scholarly journals Flower Power: Floral reversion as a viable alternative to nodal micropropagation in Cannabis sativa

2020 ◽  
Author(s):  
A.S. Monthony ◽  
S. Bagheri ◽  
Y. Zheng ◽  
A.M.P. Jones
Keyword(s):  
Tissue Culture ◽  
Large Scale ◽  
Cannabis Sativa ◽  
Insect Pests ◽  
Vegetative State ◽  
Germplasm Storage ◽  
Meristematic Tissue ◽  
Mother Plants ◽  
Photoperiod Sensitive

AbstractThe legalization of Cannabis sativa L. for recreational and medical purposes has been gaining global momentum, leading to a rise in interest in Cannabis tissue culture as growers look for large-scale solutions to germplasm storage and clean plant propagation. Mother plants used in commercial propagation are susceptible to insect pests and disease and require considerable space. While micropropagation can produce disease free starting material in less space, current published in vitro micropropagation methods are not robust and few report high multiplication rates. Further, these micropropagation methods rely on photoperiod-sensitive plants which can be maintained in a perpetual vegetative state. Current methods are not adaptable to long-term tissue culture of day-neutral cultivars, which cannot be maintained in perpetual vegetative growth. In this study, we chose to develop a micropropagation system which uses C. sativa inflorescences as starting materials. This study used two cannabis cultivars, two plant growth regulators (PGR; 6-benzylaminopurine and meta-topolin) at different concentrations, and two different numbers of florets. Here we show that floral reversion occurs from meristematic tissue in C. sativa florets and that it can be used to enhance multiplication rates compared to existing in vitro methods. Floret number was shown to have a significant impact on percent reversion, with pairs of florets reverting more frequently and producing healthier explants than single florets, while cultivar and PGR had no significant effect on percent reversion. Compared with our previously published nodal culture studies, the current floral reversion method produced up to eight times more explants per tissue culture cycle. Floral reversion provides a foundation for effective inflorescence-based micropropagation systems in C. sativa.

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