scholarly journals Improved Survival of in Vitro-stored Rubus Germplasm

1993 ◽  
Vol 118 (6) ◽  
pp. 890-895 ◽  
Author(s):  
Barbara M. Reed
Keyword(s):  
Tissue Culture ◽  
Germplasm Collection ◽  
Suitable Condition ◽  
Shoot Cultures ◽  
Medium Term ◽  
Glass Tubes ◽  
Cold Sensitive ◽  
And Storage ◽  
Plastic Tissue Culture

Medium-term in vitro cold storage of Rubus germplasm was investigated using various temperatures, photoperiods, and storage containers. Shoot cultures of several Rubus taxa were grown either in tissue-culture hags or 20 × 150-mm glass tubes. Cultures stored at 10C in darkness were in poor condition after 6 months. Overall survival and condition ratings were significantly better for bags than tubes when cultures were kept at 4C. Contamination was present in 14% of the tubes, but only 3% of the bags. Addition of a 12-hour photoperiod to 4C storage significantly improved both condition ratings and survival percentages of many individual genotypes. Evaluation of the 250-accession germplasm collection after 12 months at 4C (dark) showed 92% of accessions in bags and 85% in tubes in suitable condition to remain in storage. Storage of cold-sensitive genotypes in tissue-culture bags at 25C with a 16-hour daylength was extended to 9 months when the MS-medium nitrogen level was reduced to 25% of standard concentration. Survival of `Mandarin' raspberry stored for 9 months improved from 40% at 4C (100% N) to 90% at 25C (25% N). Results of these studies suggest that most Rubus germplasm can be stored safely at 4C with 12 hours of light. Plastic tissue-culture bags are preferred over tubes due to higher survival and lower contamination rates. Storage at 25C on reduced-nitrogen medium is an alternative method for cold-sensitive genotypes.

scholarly journals COLD STORAGE OF IN VITRO RUBUS GERMPLASM

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 695f-695
Author(s):  
Barbara M. Reed
Keyword(s):  
Tissue Culture ◽  
Cold Storage ◽  
Survival Rates ◽  
Storage Condition ◽  
Storage Conditions ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Glass Tubes ◽  
Plastic Tissue Culture

In vitro cold storage of Rubus germplasm was investigated using several environmental conditons and types of storage containers. Shoot cultures of Rubus species and cultivars were grown in either tissue culture bags or 20 × 150 mm glass tubes and compared for plant condition and survival under various storage conditions. Cultures stored at 10 C in the dark were in poor condition after 6 months. Cultures kept at 4 C were in much better condition and had higher survival rates after 18 months when stored with a 12 h daylength rather than total darkness. Overall there were no differences in survival or condition between cultures in tubes and bags. Contamination rates were 15% in tubes and 0% in bags. Plants in tissue culture bags could be stored for 9 months at 25 C with 16 h light when the nitrogen level of the MS medium was reduced to 25% and the medium volume was increased from 10 to 20 ml per bag. Genotype differences were apparent under all conditions tested. The best storage condition for Rubus germplasm was 4 C with 12 h light. Plastic tissue culture bags were preferred over tubes due to lower contamination rates.

scholarly journals Amending storage vessel and media increase transfer interval of Musa spp. tissue culture plantlets

1969 ◽  
Vol 97 (1-2) ◽  
pp. 1-14
Author(s):  
Brian M. Irish ◽  
Ricardo Goenaga ◽  
Barbara M. Reed
Keyword(s):  
Tissue Culture ◽  
Storage Time ◽  
Germplasm Collection ◽  
Monthly Basis ◽  
Musa Spp ◽  
Glass Tubes ◽  
And Storage ◽  
Field Plots ◽  
Transfer Interval

Musa spp. are some of the most important fruit food crops in the world. The USDA-ARS TARS maintains a Musa spp. germplasm collection of 164 accessions in field plots and in medium-term storage in vitro. Accessions maintained in vitro require routine sub-culturing as nutrient medium is lost due to uptake by the plant. Culture transfer intervals occur every six months and the transfer process is a resource and time consuming effort. To lengthen the transfer interval, an experiment was conducted to evaluate storage medium modifications and storage vessels on four Musa spp. accessions. Treatments consisted of glass tubes, glass tubes with Parafilm®, and plastic culture bags with three medium alterations: Murashige and Skoog (MS) medium, % strength MS and MS with 4% D-mannitol. Treatment effects were estimated by measuring plantlet's overall appearance, shoot and leaf number, and rooting on a monthly basis. All medium formulations for all four accessions, in glass tubes with Parafilm® and in culture bags showed significantly increased sub-culture interval times. The % MS treatment initially retarded plantlet development and showed the shortest storage time for all accessions. Storage time could be extended to 12 months with tissue culture bags, and to over 16 months with sealed tubes. The simplicity of using culture bags for distribution and the decrease in contamination during storage in bags were identified as additional advantages.

scholarly journals Ex Vitro Rooting of American Chestnut Improves Acclimatization Survival and Plantlet Quality

2016 ◽  
Vol 34 (3) ◽  
pp. 75-79 ◽  
Author(s):  
Allison D Oakes ◽  
Tyler R. Desmarais ◽  
William A. Powell ◽  
Charles A. Maynard
Keyword(s):  
Tissue Culture ◽  
Keystone Species ◽  
Ex Vitro Rooting ◽  
Plant Production ◽  
Shoot Cultures ◽  
Ex Vitro ◽  
Regulatory Review ◽  
Survival Percentage ◽  
Virus Indexing

Tissue culture of plants has many applications, from producing genetically identical horticultural varieties, to production of secondary metabolites, to virus indexing, and most relevantly, developing novel traits by genetic transformation. Using Agrobacterium-mediated transformation on somatic embryos, blight-resistant American chestnuts [Castanea dentata (Marsh.) Borkh.] have been developed as shoot cultures in plant tissue culture. Rooting tissue-cultured shoots and acclimatizing the rooted plantlets are key steps in tree production. In this study, in vitro and ex vitro rooting methods were compared. The ex vitro method resulted in a lower initial rooting percentage but an overall higher survival percentage, resulting in higher potted plant production. The higher survival was likely due to partial acclimatization taking place before the plantlets were transplanted into potting mix. After 8 weeks, plantlets rooted via the ex vitro method were taller, and had more, and larger, leaves than the in vitro-rooted plantlets. These trees are currently in high demand for inoculation studies for federal regulatory review and eventually for restoration of this keystone species to its native habitat.

SHOOT TISSUE CULTURE OF APPLE: COMPARATIVE RESPONSE OF FIVE CULTIVARS TO CYTOKININ AND AUXIN

1982 ◽  
Vol 62 (3) ◽  
pp. 689-694 ◽  
Author(s):  
W. D. LANE ◽  
J. M. McDOUGALD
Keyword(s):  
Tissue Culture ◽  
Acute Treatment ◽  
Shoot Cultures ◽  
Shoot Tissue ◽  
Shoot Production ◽  
Comparative Response ◽  
Apple Cultivars ◽  
Rooting Response ◽  
Scion Cultivar

Shoot cultures of five apple cultivars, M.27, M.9, M.26, MM.111 and Macspur, a strain of McIntosh, were established in vitro and their response to different concentrations of cytokinin (benzyladenine, BA) and auxin (naphtheleneacetic acid, NAA) were measured. At the three BA concentrations tested (1.0, 5.0 and 10 μM) cultivars differed in the number of shoots produced and in their requirements for BA for optimum shoot production. M.27 produced the most shoots followed by Macspur, M.9 and M.26. The best concentration of BA for shoot production was 5.0 μM for Macspur and M.26 but slightly higher for M.27 and M.9. Rooting response was tested at NAA concentrations of 0.1, 0.33, 1.0, 3.3, 10 and 33 μM. The range of concentrations in which rootstock cultivars rooted was broader than for the scion cultivar Macspur and the percent rooting of rootstocks (about 85%) was higher than Macspur (58%). The most rooting occurred at 1.0 or 3.3 μM NAA. M.9 produced callus, which prevented rooting, when chronically exposed to NAA so a procedure of acute treatment was used. This allowed root initials to form but avoided callogenesis. Possible reasons for the different responses of the cultivars to cytokinin and auxin are discussed.

scholarly journals In Vitro Storage Conditions for Mint Germplasm

HortScience ◽  
1999 ◽  
Vol 34 (2) ◽  
pp. 350-352 ◽  
Author(s):  
Barbara M. Reed
Keyword(s):  
Tissue Culture ◽  
Good Condition ◽  
Storage Conditions ◽  
Ms Medium ◽  
Mentha Arvensis ◽  
N Concentration ◽  
Temperature Regimes ◽  
Murashige And Skoog Medium ◽  
Cold Sensitive

Four tissue-cultured mints, Mentha arvensis L., M. spicata L., M. suaveolens Ehrh. hybrid, and M. suaveolens cv. Variegata, were evaluated for survival during storage in media containing three concentrations of N and in four light and temperature regimes. Shoots were placed in plastic, five-chamber, tissue-culture bags on Murashige and Skoog medium (1962) containing 25%, 50%, and 100% of the normal N concentration (MS-N) and stored at 4 °C and –1 °C in darkness, at 4 °C with a 12-hour photoperiod, and at 25 °C with a 16-hour photoperiod. Shoots of all four genotypes stored at 25 °C were in excellent condition after 6 months but required subculture after 18 months. Condition ratings of stored shoots varied with genotype and N concentration. Cultures survived longest at 4 °C with a 12-hour photoperiod on 50% MS-N. Under this regime, all four genotypes were rated in good condition at 30 months but declined to poor condition by 36 months. Based on these data, I recommend that mint cultures be stored on MS medium with 50% MS-N at 4 °C with a 12-hour photoperiod. This regime should provide a minimum of 24 to 36 months of storage before subculture is required. Cold-sensitive genotypes could be stored for 18 months at 25 °C on 50% MS-N medium.

scholarly journals In Vitro Shoot Regeneration of Georgia Plume, Elliottia racemosa, from Multiple Genotypes Collected from Wild Populations

HortScience ◽  
2011 ◽  
Vol 46 (2) ◽  
pp. 287-290
Author(s):  
Carrie A. Radcliffe ◽  
James M. Affolter ◽  
Hazel Y. Wetzstein
Keyword(s):  
Tissue Culture ◽  
Shoot Regeneration ◽  
Adventitious Shoots ◽  
Shoot Elongation ◽  
Shoot Cultures ◽  
Regeneration System ◽  
Low Genetic Diversity ◽  
Wide Range ◽  
In Vitro Shoot Regeneration

Georgia plume (Elliottia racemosa, Ericaceae) is a threatened, woody plant endemic to Georgia's Coastal Plain region in the southeastern United States. Populations of the plant have a fragmented distribution within a restricted range and are characterized by low genetic diversity and a lack of sexual recruitment. Georgia plume cannot be effectively propagated using conventional methods. We have developed an in vitro shoot regeneration system that is effective with explants obtained from mature plants in the wild. The objective of this study was to determine the efficacy of using this in vitro protocol to regenerate proliferating shoot cultures from 34 georgia plume genotypes obtained from divergent populations. Young expanding leaves were cultured on Gamborg's media supplemented with 10 μM thidiazuron and 5 μM indole-3-acetic acid. After 8 weeks, tissues were transferred to a shoot elongation medium with 25 μM 2-isopentenyl adenine. Of the 34 genotypes tested, 91% formed shoot primordia and 85% regenerated shoots within 6 months of inoculation. This study verifies that tissue culture can be used to produce adventitious shoots from a wide range of georgia plume genotypes. Within a coordinated conservation program, tissue culture is a feasible system to use for safeguarding and reintroduction of genetically diverse plant material, which may be critical to the survival of this rare species.

Comparison of Methods Used for the Removal of Dmso following Cryopreservation and the Development of an Automated Protocol

1997 ◽  
Vol 6 (2) ◽  
pp. 163-172 ◽  
Author(s):  
Jonathan R.T. Lakey ◽  
Garth L. Warnock ◽  
Ray V. Rajotte
Keyword(s):  
Tissue Culture ◽  
Low Temperature ◽  
Graft Function ◽  
T Test ◽  
Slow Step ◽  
Slow Cooling ◽  
Glass Tubes ◽  
Automated Protocol

Current methods of islet isolation are limited, thus requiring islets to be pooled from multiple donors to provide sufficient islet mass to permit insulin independence following islet transplantation. Low temperature banking is one approach used to pool islet preparations. Recently, we developed a method for bulk cryopreservation of islets in a single freezer bag system that is less labor-intensive and more readily kept sterile. As a further improvement to this bulk cryopreservation protocol we examined islet survival following slow-step dilution or our standard sucrose dilution protocol. Known numbers of canine islets were cryopreserved in DMSO by slow cooling to -40°C, storing at -196°C, and rapid thawing. When islets were frozen and thawed in glass tubes the recovery of islets after 48 h of tissue culture was significantly higher when the DMSO was removed using either a slow step (71.7 + 2.7%) or a modified slow step (75.7 + 3.9%) protocol as compared with the standard sucrose dilution protocol (65.7 + 3.0%) (p < 0.05, unpaired t-test). Insulin secretion in vitro and in vivo graft function was similar between the experimental groups. Similarly, when islets were frozen then thawed in freezer bags, islet recovery following 48 h postcryopreser-vation tissue culture at 37° C was 74.8 + 2.4% for slow-step dilution compared with 66.2 + 2.7% for the standard sucrose dilution group (p < 0.05, unpaired t-test). Islets thawed in the freezer bag using the modified slow-step dilution protocol showed equivalent functional viability during static incubation to nonfrozen controls. Bulk cryopreservation of isolated islets in single blood freezer bags is a practical alternative to cryopreservation in glass tubes. Development of an automated protocol for the slow stepwise removal of the cryoprotectant from islets in freezer bags will facilitate low temperature tissue banking of islets.

Medium-term conservation of redwood (Sequoia sempervirens (D. Don.) Endl.) in vitro shoot cultures and encapsulated buds

2011 ◽  
Vol 127 (3) ◽  
pp. 431-435 ◽  
Author(s):  
E.A. Ozudogru ◽  
E. Kirdok ◽  
E. Kaya ◽  
M. Capuana ◽  
A. De Carlo ◽  
...  
Keyword(s):  
Sequoia Sempervirens ◽  
Shoot Cultures ◽  
Medium Term ◽  
In Vitro Shoot Cultures

scholarly journals MULTIPLIKASI IN VITRO ANGGREK HITAM (Coelogyne pandurata Lindl.) PADA PERLAKUAN KOMBINASI NAA DAN BAP

2018 ◽  
Vol 5 (1) ◽  
pp. 75 ◽  
Author(s):  
Roni Kartiman ◽  
Dewi Sukma ◽  
Syarifah Iis Aisyah ◽  
Agus Purwito
Keyword(s):  
Tissue Culture ◽  
Shoot Multiplication ◽  
Basic Medium ◽  
Reproduction Rate ◽  
Shoot Cultures ◽  
Natural Reproduction ◽  
Indigenous Plant ◽  
Over Exploitation ◽  
Germinating Seeds

In Vitro Multiplication of  Black Orchid (Coelogyne pandurata Lindl.) Using the Combination of NAA and BAPABSTRACTBlack orchid is an indigenous plant from Kalimantan, Indonesia. It becomes endangered because of forest over-exploitation and its low natural reproduction rate. Tissue culture is considered to offer a solution to conserve and propagate this species. The aim of this research was to evaluate the effect of Naphtalene Acetic Acid (NAA) and 6-Benzile Amino Purine (BAP) on shoots multiplication of black orchid. The basic medium used was a half of Murashige & Skoog (MS) composition supplemented with 150 mLL-1 coconut water. Initial explants used were 6-month-old shoots of germinating seeds. The shoot cultures were incubated for 23 weeks. Results showed that the best combination for shoot multiplication was NAA 0.0 mgL-1 with BAP 0.2 mgL-1. Shoot grew better on medium with BAP and without NAA while roots growth was better on medium without the two plant growth regulators. The addition of BAP up to 0.3 mgL-1 increased the leaf number, which however decreased at higher BAP concentration.Keywords: BAP, black orchid, Coelogyne pandurata, multiplication, NAA ABSTRAKAnggrek hitam merupakan flora langka asli Kalimantan, Indonesia. Keberadaa anggrek ini di alam semakin langka akibat eksploitasi berlebihan dan sulitnya perbanyakan secara alami. Kultur jaringan merupakan metode untuk mengatasi kelangkaan anggrek ini. Penelitian ini bertujuan untuk mengetahui pengaruh kombinasi NAA dan BAP terhadap multiplikasi anggrek hitam. Media dasar yang digunakan adalah ½ MS dengan penambahan air kelapa 150 mLL-1. Eksplan yang digunakan adalah tunas hasil semai biji umur 6 bulan. Kultur tunas diinkubasi selama 23 minggu. Hasil penelitian menunjukkan bahwa kombinasi terbaik untuk multiplikasi tunas adalah NAA 0 mgL-1 dengan BAP 0,2 mgL-1. Tunas tumbuh lebih baik dalam media dengan penambahan BAP tanpa NAA, sedangkan akar pada media tanpa NAA dan BAP. Penambahan BAP sampai 0.3 mgL-1 mampu meningkatkan jumlah daun, namun menurun dengan penambahan di atas konsentrasi tersebut.Kata Kunci: anggrek hitam, BAP, Coelogyne pandurata, multiplikasi, NAA

RAPID IN VITRO PROPAGATION OF WESTERN GHATS CULTIVAR - COLOCASIA ESCULENTA FOR THE HIGH FREQUENCY OF PLANTLETS

2019 ◽  
Vol 7 (2) ◽  
Author(s):  
Jyothi R ◽  
Srinivasa Murthy K M ◽  
Hossein . ◽  
Veena .
Keyword(s):  
Tissue Culture ◽  
Wide Distribution ◽  
Colocasia Esculenta ◽  
Limiting Factor ◽  
Promising Alternative ◽  
Rapid Multiplication ◽  
Medical Plant ◽  
Total Fat ◽  
Planting Materials

Colocasia esculenta is commonly known as Taro, it is referred to as cocoyam in Nigeria. They are cherished for their rich taste, nutritional and medicinal properties. Every 100 g of taro corms possess 112 Kcal, 26.46 g carbohydrate, 1.50 g protein, 0.20 g total fat and 4.1g fiber (USDA National Nutrient Data Base). Besides its nutritional value, taro is used as a medical plant and provides bioactive compounds used as an anti-cancer drugs. Traditionally, cocoyams are vegetative propagated from tuber fragments, a practice that encourages pathogen distribution. Colocasia esculenta is a widely distributed food crop in the humid tropics and subtropics. Despite of its wide distribution, Taro plants are commonly infected with DsMV and other pathogens. This virus induces conspicuous mosaic, malformation, dwarfing or feathering on leaves in taro. As the results of infection, it reduces the quality and yield of taro production greatly. This virus is thus considered as a major limiting factor in the production of taro. Here plays the importance of  tissue culture plays a major role in producing the disease resistant plants round the year with high quality. For rapid multiplication and production of quality planting materials, tissue culture technology offers promising alternative compared to the traditional production methods. KEYWORDS: Colocasia esculenta, Virus, Pathogens, Conventional propagation, Micropropagation, Yield, Rapid multiplication, Quality

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