scholarly journals Effect of Bead Composition, PVS Type, and Recovery Medium in Cryopreservation of Bleeding Heart ‘Valentine’—Preliminary Study

Agronomy ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 891
Author(s):  
Dariusz Kulus
Keyword(s):  
Calcium Alginate ◽  
Dry Weight ◽  
Synthetic Seeds ◽  
Shoot Tips ◽  
Ms Medium ◽  
Sterile Water ◽  
Murashige And Skoog Medium ◽  
Preliminary Study ◽  
Lamprocapnos Spectabilis ◽  
Recovery Medium

Bleeding heart (Lamprocapnos spectabilis (L.) Fukuhara) is a valuable ornamental and medicinal perennial. To date, there are few studies focused on cryopreservation of this species, although it could be useful in storage and breeding. This research is aimed at analyzing the effect of bead composition, type of plant vitrification solution (PVS), and the recovery medium of cryopreservation of bleeding heart. Shoot tips of L. spectabilis ‘Valentine’ were used in the study. The explants were precultured on modified Murashige and Skoog medium (MS; 1962), supplemented with 9% sucrose, 1.0-mg·L−1 kinetin (KIN), and 2.62-mg·L−1 abscisic acid. Next, in the first experiment, the shoot tips were embedded in 3% calcium alginate, based either on an MS medium or distilled sterile water. The produced synseeds were inoculated on the recovery medium with 3.0-mg·L−1 KIN, 0.5-mg·L−1 6-benzyladenine (BA), or cytokinin–free control. Based on the results of the first study, in the second experiment, precultured shoot tips were embedded in 3% calcium alginate based on MS medium and dehydrated with PVS2 or PVS3 for various durations. The pre-treated explants were plunged in liquid nitrogen and, after rewarming, inoculated on the recovery MS medium with 0.5-mg·L−1 BA. PVS3 was more effective in securing the shoot tips than PVS2. The highest recovery level (68.3%) was reported after a 150-min pretreatment with PVS3. Explants from this experimental combination also proliferated the highest number of shoots, as well as those with the greatest length. On the other hand, a higher share of dry weight was found in PVS2-derived shoots (13.5–18.2%) compared with plants produced after PVS3 treatment (10.6–11.4%). The obtained results here can serve as a good basis for further studies related to synthetic seeds and cryopreservation of bleeding heart.

scholarly journals Gold nanoparticles affect the cryopreservation efficiency of in vitro-derived shoot tips of bleeding heart

2021 ◽  
Author(s):  
Dariusz Kulus ◽  
Alicja Tymoszuk
Keyword(s):  
Gold Nanoparticles ◽  
Alginate Bead ◽  
Stability Index ◽  
Shoot Tips ◽  
Start Codon ◽  
Lamprocapnos Spectabilis ◽  
Colloid Dispersion ◽  
Recovery Medium ◽  
Alginate Matrix

AbstractThe popularity of nanoparticles (NPs) is continuously increasing. To date, however, there has been little research on the application of NPs in plant cryopreservation, i.e. storage of tissues in liquid nitrogen (LN). The aim of this study is to analyze the effect and evaluate the usefulness of gold nanoparticles (AuNPs) in regard to cryobiology studies. In vitro-derived shoot tips of Lamprocapnos spectabilis ‘Valentine’ were cryopreserved with the encapsulation-vitrification protocol. Gold nanoparticles (at 10–30 ppm concentration; 13 nm in size) were added either into the preculture medium; to the protective bead matrix during encapsulation; or to the recovery medium after rewarming of samples. The control plants were produced from cryopreserved explants non-treated with nanoparticles or treated with colloid dispersion medium without NPs. A non-LN-treated standard was also considered. The influence of AuNPs on the cryopreservation efficiency was determined by evaluating the recovery rate of explants and their morphogenic response; the membrane stability index (MSI); the concentration of pigments in shoots; and the antioxidant enzymes activity. The genetic stability of the plant material was evaluated using Start Codon Targeted Polymorphism (SCoT) markers. It was found that 10 ppm of AuNPs added into the alginate bead matrix improved the recovery level of LN-derived shoot tips (70.0%) compared to the non-NPs-treated cryopreserved control (50.5%). On the other hand, the presence of nanoparticles in the recovery medium had a deleterious effect on the survival of explants. AuNPs usually had no impact on the MSI (73.9–85.9%), except for those added into the recovery medium at the concentration of 30 ppm (decline to 55.8%). All LN-derived shoots were shorter and contained less chlorophyll and carotenoids than the untreated standard. Moreover, the application of AuNPs affected the enzymatic activity in L. spectabilis. Minor genetic variation was found in 8.6% of plants if AuNPs were added either into the preculture medium (at 10 and 20 ppm) or to the alginate matrix (at 30 ppm). In conclusion, AuNPs added at a lower concentration (10 ppm) into the protective bead matrix can significantly improve the cryopreservation efficiency in L. spectabilis with no alternation in the DNA sequence.

scholarly journals Optimation of Auxin and Cytokinin on Enhanced Quality and Weight of Coffea liberica Somatic Embryos

2021 ◽  
Vol 37 (1) ◽  
Author(s):  
Fitria Ardiyani ◽  
Edy Setiti Wida Utami ◽  
Hery Purnobasuki
Keyword(s):  
Somatic Embryo ◽  
Somatic Embryos ◽  
Dry Weight ◽  
Development Stage ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Skoog Medium ◽  
Murashige And Skoog Medium ◽  
Coffea Liberica ◽  
Germination Stage

Coffea liberica is a variety of coffee that tolerant to marginal land, especially peatlands. One of propagation methods in C. liberica is somatic embryogenesis(SE) which producing large number of true-to-type plant seedlings in a short time. This research aimed at studying the effect of application of plant growthregulator (PGR) on quality and weight of somatic embryo of C. liberica. Somatic embryo in development stage was induced by Murashige and Skoog medium containing cytokinin as benzyl amino purin (BAP) and auxin as 2,4-dichlorophe-noxyacetic acid (2,4-D). While cotyledonary embryo in germination stage was induced by Murashige and Skoog medium containing cytokinin (BAP) and auxins as 2,4-D, indole acetic acid (IAA) and naphthaleneacetic acid (NAA). The resultsshowed that the application of auxins and cytokinins on development stage affected the formation of embryos, texture of calli, color of calli and embryos, and weight of somatic embryo. It also influenced the shoot and root formation, color and weight of geminating embryos of C. liberica at the germinating stage. During the development stage, addition of 1 mg/L BAP in the absence of 2,4-D in MS medium produced the highest quality of somatic embryo of C. liberica. This medium also produced heaviest somatic embryos but with lighter callus. While in germination stage, all medium treatments produced a typical germinating embryo. Coffea liberica germinating embryo growth optimally on MS medium containing 0.5 mg/L BAP as a single chemical or 0.5 mg/L BAP in combination with 0.5 mg/L IAA for shooting development. Whereas on rooting development, addition of 0.5 mg/L NAA on MS medium produced an optimal germinating embryo. Moreover, germination embryo of C. liberica recorded the highest in terms of dry weight on MS media with addition of 0.5 mg/L BAP. Application of appropriate concentration of auxin and cytokinin is needed to support the formation of somatic embryo and germinating embryo.

scholarly journals Callus Induction and Synthetic Seed Development in Draceana sanderiana Sanderex Mast: Lucky Bamboo

2019 ◽  
pp. 1-8
Author(s):  
Mohmad Amin ◽  
Abdul Mujeeb
Keyword(s):  
Sodium Alginate ◽  
Seed Development ◽  
Callus Induction ◽  
Callus Formation ◽  
Synthetic Seeds ◽  
Shoot Tips ◽  
Synthetic Seed ◽  
Ms Medium ◽  
Seed Formation ◽  
Light Yellow

The study was carried out for callus induction and synthetic seed development from the shoot tips of Draceana sanderiana sander ex Mast. The shoot tips were subjected to different concentrations (0.25, 0.5 &1.0 mg/l) of 2,4-D on MS medium. The research findings revealed that the 2,4-D at concentrations of 0.25 mg/l was more suited for the profuse callus formation. The friable and light yellow callus was induced within 2 weeks of culture at 0.25 mg/l of 2,4-D on MS medium as compared to the other two concentrations of 2,4-D i.e.; 0.5 and 1.0 mg/l. Similarly the effect of sodium alginate and calcium chloride percentage on synthetic seed formation was observed, it was found that somatic embryos formed from shoot tips via callus kept in 2.5% sodium alginate and 100 milli molar CaCl2 produced synthetic seeds with firm spherical beads. The study leads to the formation of synthetic seeds of Draceana sanderiana which can be used for the conservation of germplasm through cryopreservation and the micro propagation of the said plant species.

scholarly journals Preparation of Synthetic Seeds of Citrus jambhiri Using in vitro Regenerated Multiple Plantlets

2020 ◽  
pp. 22-29
Author(s):  
Priyanka Sharma ◽  
Bidhan Roy
Keyword(s):  
Germplasm Conservation ◽  
Synthetic Seeds ◽  
Shoot Tips ◽  
Nodal Segments ◽  
Synthetic Seed ◽  
Ms Medium ◽  
Meristematic Tissue ◽  
Citrus Jambhiri ◽  
Cacl2 Solution

Biotechnological tools are useful for true-to-type propagation. Shoot tips encapsulation is potential for plant development from pre-existing meristematic tissue. MS medium fortified with 1 and 2 mg/L of BAP (6-bezylaminopurine) was found to be suitable for in vitro mass-multiplication of plantlets (10.18 and 13.05 plantlets/explant, respectively) of Citrus jambhiri from nodal segments. Nodal segments were more appropriate than the shoot tips for in vitro multiplication of plantlets. Synthetic seeds were prepared using 2.5% sodium alginate dropping into 3.0% CaCl2 solution. Maximum germination was recorded when beaded shoot tips were cultured on MS medium fortified with 1 and 2 mg/L of BAP (96.67 and 100.00%, respectively). However, the germination of synthetic seeds was found to be comparatively high than the earlier findings. The results support the use of encapsulated unipolar explants for synthetic seed preparation. These type of capsules could be useful in exchange of sterile material between laboratories, germplasm conservation and direct plant propagation.

scholarly journals Micropropagtion of two Strawberries (Fragaria anariassa L.) Cultivars

2008 ◽  
Vol 2 (1) ◽  
pp. 42-50
Author(s):  
Dhaher Abas ◽  
Jabbar abas Hassan ◽  
Abd aljasem. M.Jasem
Keyword(s):  
Nutrient Medium ◽  
Dry Weight ◽  
Shoot Tips ◽  
Peat Moss ◽  
Ms Medium ◽  
Fresh Weight

Shoot tips from two strawberry (Fragaria anariassa L.) cultivars, Gabi and Edpeerenwere cultured on MS medium supplemented with 0, 0.5, 1.0, 1.5 or 2.0mg /1 of BA tostudy their effects on number, length, fresh weight and dry weight of shoots. Thestudy also included measuring the effect of different concentrations of IBA (0 , 0.1 ,0.3 , 0.5 or 1.0) mg / l on rooting percentage, roots number and length. This studyalso included the acclimatization processes for plantlets in sand and a mixture ofsand and peat moss media. Results indicated that addition of BA to the nutrientmedium led to a significant increase in all investigated traits in comparison with the control with obvious superiority of 2 mg /l which reflected positively on the meannumber of shoots (20.0) shoots / plant, the fresh and dry weight were 1237.73 and136.4 mg respectively, especially with Gabi cultivar, the result showed that all thelevels of IBA in the nutrient medium led to 100% rooting after 6 weeks of plantingof both cultivars compared with 50% of in control. It was also noticed significantincreases in the roots number and shoot lengths when IBA concentration wasincreased in the medium. The best growth for the rooted plantlets was found to be1:1 sand: peat moss which showed survival percentages of 80 and 90% after 4weeksof planting Erdpeern and Gabi cultivars respectively.

scholarly journals Critical vitrification steps towards survival of Garcinia hombroniana (Clusiaceae) shoot tips after cryopreservation

2018 ◽  
Vol 66 (3) ◽  
pp. 1314
Author(s):  
Norafarain Sulong ◽  
Nurul Farhana Shahabudin ◽  
Normah Mohd Noor
Keyword(s):  
Water Content ◽  
Liquid Nitrogen ◽  
Shoot Tips ◽  
The Other ◽  
Skoog Medium ◽  
Murashige And Skoog Medium ◽  
Vitrification Solution ◽  
Cryopreservation Protocol ◽  
Recovery Medium

A cryopreservation protocol was developed for in vitro shoot tips of Garcinia hombroniana using the vitrification technique. Four critical steps in the technique were investigated, namely preculture, loading, dehydration with Plant Vitrification Solution 2 (PVS2), and unloading. Shoot tips precultured for 48 hr gave significantly higher survival (75 %) compared to 24 hr preculture (50 %) after cryopreservation. Treatment with 1 M glycerol plus 0.4 M sucrose as a loading solution gave higher survival (45.83 %) compared to the other treatments (0.4 M sucrose + 2 M glycerol; 0.4 M sucrose). Shoot tips dehydrated with PVS2 for 25 min gave the highest survival after immersion in liquid nitrogen. Stepwise PVS2 treatment for 15 min with 50 % PVS2 followed by 10 min with 100 % PVS2 solution improved survival of the shoot tips after cryopreservation (41.67 %). Murashige and Skoog medium with 0.4 M sucrose gave significantly higher survival (66.67 %) than MS with 1.2 M sucrose (25 %) as an unloading solution. Water content was shown to decrease throughout the whole vitrification steps from 6.83 ± 1.66 g g-1 dw for fresh shoot tips down to 2.93 ± 0.28 g g-1 dw after PVS2 treatment. Further study on each step including recovery medium is required to improve the survival. Nevertheless, the present study showed the potential of using the vitrification technique for cryopreservation of G. hombroniana.

scholarly journals Effect of N-methyl-N-nitro-N-nitrosoguanidine (NTG) on callus induction and survival rate on mature embryos of beans (Phaseolus vulgaris)

2009 ◽  
Vol 3 (2) ◽  
pp. 91-98
Author(s):  
Sattar A. Shlahi ◽  
Zahra N. Hashim Al- Hattab
Keyword(s):  
Survival Rate ◽  
Callus Induction ◽  
Indole Acetic Acid ◽  
Casein Hydrolysate ◽  
Dry Weight ◽  
Ms Medium ◽  
Fresh Weight ◽  
Benzyl Adenine ◽  
Mature Embryos ◽  
Survival Percentage

This research was conducted to study the effect of the chemical mutagen N-methyl-N-nitro-N-nitrosoguanidine on the percentage of callus induction and survival from mature beans embryos harvester cultivar. Seeds were treated with (0.2 or 0.4) millimolar of the mutagen NTG in combination with 0.0, 4 or 8% of ethanol, pH 5 ±2 0. for 24 h. Calli were induced on mature embryos by using MS medium with 0.5 mg/l of Benzyl adenine (BA), 1 mg/l Indole acetic acid (IAA) and 100 mg/l from each of Casein hydrolysate, Glycine, Asparagine, Tyrosine, and Myo-Inositol. Results showed that the hypocotyl surpassed the radical and the plume significantly in terms of survival reached 56.3%. Mutagen treatments showed asignificant effect on calli survival. Treatment with 8% Ethanol was lethal for all explants. While treatment with 0.4 mM NTG without Ethanol gaved the highest survival rate. The interaction between the treatments and the explants showed that the lowest survival percentage was which 8.8% that was for shoots treated with 0.2 mM of 4% Ethanol. Calli induced on hypocotyls treated with 0.4 mM NTG without Ethanol gave the highest fresh weight (347.2) mg while the lowest was (60) mg for calli induced on the radical treated with 0.4 mM NTG with 4% Ethanol. Moreover the highest dry weight was 22.5 mg for calli induced from hypocotyls treated with 0.4 millimolar NTG without Ethanol that was higher than the control 17.2 mg.The lowest dry weight obtained from calli induced on the radical treated with 0.4 mM NTG with 4% Ethanol was 3 mg. In conclusion the results showed that 0.4 mM NTG without Ethanol gave the highest survival rate and the highest fresh and dry weight for calli induced on the hypocotyl.

scholarly journals Stilbenes in the different organs of Vitis vinifera cv. Merlot grafted on Teleki Kober 5BB rootstock

OENO One ◽  
2017 ◽  
Vol 51 (3) ◽  
pp. 323 ◽  
Author(s):  
Gyöngyi Németh ◽  
Orsolya Hegyi ◽  
Attila Dunai ◽  
Laszlo Kocsis
Keyword(s):  
Vitis Vinifera ◽  
Field Study ◽  
Primary Source ◽  
Dry Weight ◽  
Shoot Tips ◽  
Font Size ◽  
Plant Organs ◽  
Wide Range ◽  
Berry Skins ◽  
Winter Buds

<p style="text-align: justify;"><span style="font-size: medium;"><strong>Aim: </strong>To determine which of the grapevine organ is the richest in stilbene, like the t-piceid, t-resveratrol and ε-viniferin and would it be useful as by-product of viticulture for processing stilbene concentrate.</span></p><p style="text-align: justify;"><span style="font-size: medium;"><strong>Methods and results: </strong>Nine organs, such as cane, buds, shoot tips, inflorescences, clusters at veraison, matured berry skins, seeds and cluster stems, of Merlot grafted on Teleki Kober 5BB rootstock were collected during a year and its ethanolic extractions were analyzed via HPLC-FLD. Stilbene content of the different organs occurred in a wide range, the lowest was 3.15 mg/kg dry weight (dw) measured in the seeds and the highest 2265 mg/kg dw in the buds.</span></p><p style="text-align: justify;"><span style="font-size: medium;"><strong>Conclusion:</strong> The present research demonstrated that different grapevine organs contain different amount of stilbene in the vineyard. The winter-buds, the roots and the matured cane internodes of Merlot are significantly richer in t-resveratrol and t-ε-viniferin than the green vegetative and generative parts we examined in one growing year. </span></p><p style="text-align: justify;"><span style="font-size: medium;"><strong>Significance and impact of the study: </strong>According to our knowledge there was no stilbenes measuring in several plant organs of a vine cultivar in field study, in vineyard. Our research pointed out the pruned cane could be primary source of stilbene of the health-industry. Beside that grape root is a by-product of grapevine nursery is also rich in stilbene. </span></p>

scholarly journals Comparative study on meristem culture of three potato cultivars diamant, cardinal and granula and their shoot formation

2016 ◽  
Vol 1 (3) ◽  
pp. 537-544
Author(s):  
Marufa Khatun ◽  
Md Shahadat Hossain ◽  
Md Khalekuzzan ◽  
Amina Rownaq
Keyword(s):  
Comparative Study ◽  
Growth Response ◽  
Potato Cultivar ◽  
Shoot Formation ◽  
Shoot Tips ◽  
Potato Cultivars ◽  
Meristem Culture ◽  
Ms Medium ◽  
Surface Sterilization ◽  
Apical Meristems

The main aim of this study was to analyze the comparative study on meristem culture of three important potato cultivars, diamant, cardinal and granula and their shoot formation using meristem tips. Apical meristems were isolated from shoot tips of 25-35 days old field grown plants. After surface sterilization the meristems (0.3 mm) were isolated from the shoot tips. After isolation the apical meristems were placed quickly on “M” shaped filter paper bridge in culture tubes containing liquid MS medium supplemented with various concentrations and combinations of different plant growth regulators. Meristems showed their first growth response by increase in size and became greenish white in colour. They continued their growth and developed shoots with roots. In the present investigation, among the three cultivars of potato, cardinal were the best responsive cultivars for the resuming new growth of cultured meristems on MS medium supplemented with KIN (0.4 mg l-1) + GA3 (0.5 mg l-1). Cultivar diamant showed comparatively better growth response in the primary culture and granula showed better results in shoot length formation in MS0 medium. Cultivar cardinal proved to be best potato cultivar in case of meristem culture than diamant and granula cultivars.Asian J. Med. Biol. Res. December 2015, 1(3): 537-544

scholarly journals Adjustment of Mineral Elements in the Culture Medium for the Micropropagation of Three Vriesea Bromeliads from the Brazilian Atlantic Forest: The Importance of Calcium

HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 106-112 ◽  
Author(s):  
Alice Noemí Aranda-Peres ◽  
Lázaro Eustáquio Pereira Peres ◽  
Edson Namita Higashi ◽  
Adriana Pinheiro Martinelli
Keyword(s):  
New Media ◽  
Mineral Composition ◽  
Culture Media ◽  
Dry Weight ◽  
Defined Medium ◽  
Ms Medium ◽  
Media Formulation ◽  
Half Strength

Many different species of Bromeliaceae are endangered and their conservation requires specific knowledge of their growth habits and propagation. In vitro culture of bromeliads is an important method for efficient clonal propagation and in vitro seed germination can be used to maintain genetic variability. The present work aims to evaluate the in vitro growth and nutrient concentration in leaves of the epiphyte bromeliads Vriesea friburguensis Mez, Vriesea hieroglyphica (Carrière) E. Morren, and Vriesea unilateralis Mez, which exhibit slow rates of growth in vivo and in vitro. Initially, we compared the endogenous mineral composition of bromeliad plantlets grown in half-strength Murashige and Skoog (MS) medium and the mineral composition considered adequate in the literature. This approach suggested that calcium (Ca) is a critical nutrient and this was considered for new media formulation. Three new culture media were defined in which the main changes to half-strength MS medium were an increase in Ca, magnesium, sulfur, copper, and chloride and a decrease in iron, maintaining the nitrate:ammonium rate at ≈2:1. The main difference among the three new media formulated was Ca concentration, which varied from 1.5 mm in half-strength MS to 3.0, 6.0, and 12 mm in M2, M3, and M4 media, respectively. Consistently, all three species exhibited significantly higher fresh and dry weight on M4, the newly defined medium with the highest level of Ca (12 mm). Leaf nitrogen, potassium, zinc, magnesium, and boron concentrations increased as Ca concentration in the medium increased from 1.5 to 12 mm.

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