scholarly journals Kriopreservasi Tunas in Vitro Pepaya ‘Sukma’ Dengan Perlakuan Prakultur, Loading, Dan Dehidrasi Dengan Pvs2 Dan Modifikasinya

2021 ◽  
Vol 20 (2) ◽  
pp. 181-190
Author(s):  
Fitri Fatma Wardani ◽  
Joko Ridho Witono ◽  
Darda Efendi ◽  
Diny Dinarti
Keyword(s):  
Survival Rate ◽  
Genetic Variability ◽  
Liquid Nitrogen ◽  
Treatment Time ◽  
Ms Medium ◽  
High Genetic Variability ◽  
Environment Changes ◽  
In Vitro Shoots

Papaya has high genetic variability because it is an open-pollinated plant and has genotype and phenotypeare that are easily changed due to environment changes. Cryopreservation is a storing method of germplasm in liquid nitrogen (-196 oC) which can maintain the genotype and phenotype of germplasm. The experiment aimed to obtain the best preculture, loading, and dehydration for cryopreservation of papaya ‘Sukma’ in vitro shoots. For preculture, we planted shoots on MS media with 0.3 M and 0.4 M sucrose for 1, 2, and 3 days. In the loading treatment, we immersed shoots in loading solution (liquid MS+1.2M glycerol+0.4M sucrose) for 0, 10, 20, and 30 minutes. For dehydration, we immersed shoots in cryoprotectant (PVS2 and its modification) for 5, 10, and 15 minutes. Then, shoots were immersed in liquid nitrogen. The results showed thatshoots had the best survival rate while they had been precultured on MS medium with 0.3 M sucrose for 3 days. The best loading treatment time was 20–30 minutes. The best dehydration treatment was obtained by modification of PVS2 for 10 minutes. The shoots have not been able to recovery after cryopreservation, so it can be concluded that cryopreservation of in vitro papaya ‘Sukma’ shoots has not been successful.

scholarly journals In vitro Shoot Cultures of Tupistra albiflora K. Larsen

2018 ◽  
Vol 17 (5) ◽  
pp. 405-411
Author(s):  
Jiraporn PALEE
Keyword(s):  
Agar Medium ◽  
Multiple Shoots ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Naphthalene Acetic Acid ◽  
Root Induction ◽  
Shoot Cultures ◽  
Ms Medium ◽  
In Vitro Shoots

To evaluate an efficient protocol for the micropropagation of Tupistra albiflora K. Larsen, the effects of N6-benzylaminopurine (BA) and naphthalene acetic acid (NAA) concentrations on multiple shoot and root induction were examined. In vitro shoots were used as the explant materials which were cultured on Murashige and Skoog (MS) agar medium supplemented with 0, 1, 2, 3 and 4 mg/L BA for 4 weeks to induce multiple shoots. It was found that the MS medium containing 3 mg/L BA induced 100 % shoot formation with the highest number of 3.2 shoots per explant (2.4-fold significantly higher than the control). For root induction, in vitro shoots were cultured on MS agar medium supplemented with 0, 1, 2, 3 and 4 mg/L NAA for 8 weeks. The results showed that the MS medium containing 1 mg/L NAA induced 100 % root formation with the highest number of 6.6 roots per explant (1.8-fold significantly higher than the control).

scholarly journals Induksi PLB Anggrek Vanda sumatrana Schltr. Liar Pada Media MS dengan Penambahan BAP dan NAA serta Ploidisasi dengan Kolkisin

2015 ◽  
Vol 4 (4) ◽  
pp. 208
Author(s):  
Hanifah - Aini ◽  
Mansyurdin - Mansyurdin ◽  
Suwirmen - Suwirmen
Keyword(s):  
Plant Physiology ◽  
Tissue Culture ◽  
Acetic Acid ◽  
Survival Rate ◽  
Cell Biology ◽  
Colchicine Treatment ◽  
Ms Medium ◽  
Colchicine Solution ◽  
Department Faculty

The study about PLB induction of wild Vanda sumatrana Schltr. on MS media suplement with BAP and NAA and ploidisation by colchicine treatment was conducted from December 2014 until November 2015 at the Laboratory of Genetics and Cell Biology and Laboratory of Plant Physiology and Tissue Culture, Biology department, Faculty of Mathematic and Natural Science, Andalas University, Padang. The study aimed to 1) knowing the best concentration of 6-Benzyl amino purin (BAP) and α-Naphtalene acetic acid (NAA) for Protocorm Like Bodies (PLB)  induction from shoot tip of V. sumatrana, 2) knowing the PLB response of V. sumatrana to concentrations and soak period of colchicine and 3) find the effective concentrations and soak period of colchicine to induce tetraploid on PLB of V. sumatrana. Shoot tips from in-vitro cultured of V. sumatrana  were subcultured on Murashinge and Skoog (MS) medium supplement with 3 mg/l BAP + 0,5 mg/l NAA, 3 mg/l BAP and 1,5 mg/l BAP. PLB of diploid V. sumatrana from the best treatment were soaked in 0.05% and 0.1% colchicine for 24 and 48 hours respectively in MS liquid medium, as control were set PLB without colchicine treatment. The results showed that MS medium supplemented with 1.5 mg/l BAP was the best formula to induce PLB. The highest percentage of survival rate of PLB and percentage of survived PLB regenerated shoot was obtained from 0.05% colchicine with 24 hours soak period treatment. The effective treatment to induce tetraploid on PLB of V. sumatrana Schltr. was obtained from 0.05% colchicine solution for 24 hours soak period.

scholarly journals Propagation of Dendrobium aggregatum by green capsule culture

Lankesteriana ◽  
2013 ◽  
Author(s):  
S. Vijayakumar ◽  
G. Rajalkshmi ◽  
K. Kalimuthu
Keyword(s):  
Root Formation ◽  
Coconut Water ◽  
Immature Embryos ◽  
Ms Medium ◽  
Medium Supplement ◽  
In Vitro Shoots ◽  
Immature Seeds ◽  
Benzyl Amino Purine ◽  
Number Of Shoots

An efficient protocol for propagation of Dendrobium aggregatum using the axenic immature seeds, derived from green capsule, was developed. The immature embryos from 120 days old capsules after pollination were germinated on Murashige and Skoog (MS) medium supplement with various concentration of BAP alone or in combination with NAA along with coconut water, and the same media were used for induction, multiplication, elongation and rooting in vitro shoots. MS medium with the addition of 3% sucrose 1.5 mg L-1 Benzyl amino purine (BAP) and 15% coconut water (CW) favoured the higher rate of germination, more number of protocorm like bodies, production of maximum number of shoots, elongation of shoots, as well as root formation. During acclimatization, 95% of the plantlets survived after one month.

scholarly journals The in vitro shoot regeneration of Ehretia asperula Zol. and Mor.

2019 ◽  
Vol 2 (6) ◽  
pp. 75-83
Author(s):  
Le Thi Thuy Tien
Keyword(s):  
Woody Plant ◽  
Ms Medium ◽  
Benzyl Adenine ◽  
Shoot Initiation ◽  
In Vitro Shoots ◽  
Woody Plant Medium ◽  
Number Of Leaves ◽  
Growth Experiments ◽  
In Vitro Shoot Regeneration

Xa den young branches in the orchard were sterilized and used as explants for shoot initiation and growth experiments. The shoot induction was carried out with BA (benzyl adenine) or TDZ (thidiazuron). New shoots sprouted after one week of culture and the highest shoots (2.02 cm) were on MS medium (Murashige and Skoog medium) with BA 0.6 mg/L after 3 weeks. Furthermore, the number of leaves per shoot was also higher than other treatments (8.31 leaves per shoot). In vitro shoots were used in other experiments to investigate the effects of explants, biotin concentrations, minerals, type and concentration of cytokinins on the formation and elongation of shoots and clusters. When 5 mg/L biotin was added to MS medium, shoots grew better. The MS medium appeared to be most suitable for the initiation and elongating of shoots, followed by WPM (woody plant medium) and SH medium (Schenk and Hildebrandt medium), while B5 medium (Gamborg B5 medium) was the least effective. The spouting from three-week-old explants was earlier than others (4 and 5 weeks of age), which in turn affected on the shoot elongating. BA 1.5 mg/L was suitable to induce shoot clusters (3.91 shoots per explant) after 4 weeks.

scholarly journals Salinity-Induced Alterations in the Growth, Membranes and Osmolytes of Musa spp. cv. Caipira (AAA) During Micropropagation

2019 ◽  
Vol 11 (6) ◽  
pp. 171
Author(s):  
Yuri Lima Melo ◽  
Isabele Aragão Gomes Trindade ◽  
Monique Cristina Simão Lopes ◽  
Cibelley Vanúcia Santana Dantas ◽  
Josemir Moura Maia ◽  
...  
Keyword(s):  
Survival Rate ◽  
Plant Height ◽  
Exposure Time ◽  
Root Length ◽  
Growth Traits ◽  
In Vitro Rooting ◽  
Ms Medium ◽  
Musa Spp ◽  
Number Of Leaves

The aim of this study was to determine the concentration and exposure time to NaCl suitable for the micropropagation of banana, through the analysis of growth traits. Banana propagules were inoculated in MS medium with different concentrations of NaCl (0; 50; 75 and 100 mM) for 120 days (multiplication and rooting, 60 days each), with monthly subcultures. These propagules were measured for plant height, number of leaves, sprouting rate, average number of formed propagules, rooting rate, root length and survival rate. After 30 days, NaCl reduced sprouting rate at multiplication; the number of leaves, rooting rate and root length in rooting; and the height and propagules number in both phases. After 60 days, the NaCl affected the sprouting rate and propagules number in the multiplication; length of root in rooting; and the height and number of leaves in both phases. After 120 days, the reduction in the survival rate was proportional to the increase of NaCl in the medium. Thus, it is concluded that NaCl reduces most of the growth traits and the treatments with 75 and 100 mM NaCl affected multiplication and in vitro rooting more intensely.

scholarly journals Development of Shoot Cultures from Leaf Explant of Portulaca quadrifida L.

2019 ◽  
Vol 11 (1) ◽  
pp. 45-50 ◽  
Author(s):  
Ashutosh PATHAK ◽  
Aruna JOSHI ◽  
Asha SHARMA
Keyword(s):  
Acetic Acid ◽  
Carbon Sources ◽  
Leaf Explants ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Medicinal Value ◽  
In Vitro Shoots ◽  
Shoot Number ◽  
Response Variation

Portulaca quadrifida (Portulacaceae) is an annual succulent herb having medicinal value and is consumed as a vegetable or salads in India. In the present study, leaf explants were inoculated on Murashige and Skoog’s (MS) medium fortified with sucrose (3%) and combinations of N6-benzyladenine (6-BA) and kinetin (KIN) individually and in combination with 1-naphtalene acetic acid (NAA). Rapid regeneration was observed in medium fortified with combinations of 6-BA (8 µM) and NAA (1 µM) which formed 19.40 ± 0.64 shoots with 100% response. Variation in sucrose concentrations (4-6%) was tried but it failed to increase the shoot number. When the optimized medium was fortified with different carbon sources viz. dextrose, glucose and maltose, they could not evoked better response and sucrose proved to be more effective for regeneration. Rooting of in vitro shoots was achieved in ½MS + sucrose (1%) + indole-3-butyric acid (IBA, 2 µM).

114 CRYOTOP VITRIFICATION FOR IN VITRO-PRODUCED BUFFALO (BUBALUS BUBALIS) EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 174 ◽  
Author(s):  
A. De Rosa ◽  
L. Attanasio ◽  
L. Boccia ◽  
G. Pellerano ◽  
G. Campanile ◽  
...  
Keyword(s):  
Survival Rate ◽  
Liquid Nitrogen ◽  
Sucrose Solution ◽  
Bubalus Bubalis ◽  
Thin Strip ◽  
Embryo Survival ◽  
Vitro Fertilization ◽  
Group A ◽  
Group B

The aim of this study was to compare the efficiency of different combinations of cryoprotectants for vitrification of IVP buffalo (Bubalus bubalis) embryos by the cryotop method (Kuwayama et al. 2005 RBM Online 11, 300–308). In group A, we evaluated the vitrification and warming solutions previously used to vitrify buffalo embryos in French straws (Gasparrini et al. 2001 Theriogenology 55, 307). Embryos were equilibrated in 1.4 M glycerol for 5 min and in 1.4 M glycerol and 3.6 M ethylene glycol (EG) for an additional 5 min. After being transferred into 3.4 M glycerol and 4.6 M EG for 25 s, individual embryos were picked up in an extremely small volume (<0.1 �L) of vitrification solution and placed on the top of a very fine polypropylene strip (0.4 mm wide � 20 mm long � 0.1 mm thick) attached to a hard plastic handle, kindly provided by M. Kuwayama. Each embryo was placed onto the thin strip of the Cryotop and immediately submerged into liquid nitrogen. For warming, the strip of the Cryotop was immersed directly into a 0.5 M sucrose solution; embryos were retrieved and transferred into 0.25 M sucrose for 5 min before culture in SOF medium. In group B, we examined the vitrification and warming solutions previously used for OPS vitrification of buffalo embryos (De Rosa et al. 2006 Reprod. Fertil. Dev. 18, 153). Embryos were equilibrated in 7.5% EG + 7.5% dimethyl sulfoxide (DMSO) for 3 min before transfer into 16.5% EG + 16.5% DMSO + 0.5 M sucrose. After 25 s, they were placed on the cryotop, as previously described, and submerged into liquid nitrogen. For warming, embryos were recovered into a 0.25 M sucrose solution for 1 min, transferred into 0.15 M sucrose for 5 min, and cultured in SOF. IVP buffalo embryos of excellent quality that, by Day 7 of culture (Day 0 = in vitro fertilization), had reached the blastocyst stage (n = 44 and 53 for groups A and B, respectively), over 6 replicates, were vitrified. Embryo survival rate was determined as the percentage of vitrified-warmed embryos undergoing further development during a 24-h in vitro culture period. Differences between methods were analyzed by chi-square test. A significantly higher embryo survival rate was recorded in Group B compared to Group A (67.9 vs. 43.2% respectively; P < 0.05). In conclusion, it was demonstrated that cryotop vitrification, with the combination of cryoprotectants used in group B, is a valid tool to cryopreserve IVP buffalo blastocysts.

scholarly journals Clonal Micropropagation of representatives of the genus Dactylorhiza Neck. ex Nevski

2021 ◽  
Vol 4 (46) ◽  
pp. 17-17
Author(s):  
Alexander Saakian ◽  
◽  
Keyword(s):  
Survival Rate ◽  
Growth Regulators ◽  
In Vitro Propagation ◽  
Culture Medium ◽  
Culture Media ◽  
Ms Medium ◽  
Organic Additives ◽  
Clonal Micropropagation ◽  
Half Strength

Abstract The aim of this study is to develop and improve methods of in vitro propagation of representatives of Dactylorhiza: D.baltica , D. fuchsii. For the study, we used protocorms obtained by the asymbiotic germination of seed during 90 days. It has been established that half-strength of Murashige and Skoog (1962) medium (½ MS) supplemented with 1-2 mg/l 6-Benzylaminopurine(6-BAP), potato puree (20g/l), and charcoal (1g/l) effectively influenced the development of protocorms, and seedlings formation in the studied species. The result of the study showed that the survival rate of protocorms was high in all experimental culture media, but in D. fuchsii it was better at a concentration 2mg/l of 6-BAP (95.4%), while in D. baltica it was high at 1mg/l (87.0%). The highest percentage of multiple protocorms (68%) and the formation of new secondary protocorms in D. fuchsii (5,5±0,3 units) were observed on a culture medium containing 2 mg/l 6-BAP. The highest percent of rooting of D. fuchsii protosoms (78%) and length of roots (0.9cm) observed in ½ MS medium without growth regulators. During the development of D. baltica protosoms, the culture medium of ½ MS containing 1 mg/l 6-BAP had the best effect on the number of roots (1.8±0.1root/protosom), while the medium supplemented with 2mg/l of 6-BAP contributed to the formation of a larger number of new secondary protocorms (3,2±0,1protocorm/unit). During the subsequent cultivation of protosoms of D. baltica on a culture medium containing 1 mg/l it was observed an increase in the height of shoots (4,8±0,3 см), and the length of roots (2,2±0,1 см), wherein the number of newly formed protocorms was higher by 30% on the medium supplemented with 2 mg/l 6-BAP. Keywords: DACTYLORHIZA BALTICA, DACTYLORHIZA FUCHSII, IN VITRO, PROTOCORMS, ORGANIC ADDITIVES

scholarly journals Plant Regeneration from Protocorm Like Body (PLB) derived Callus of Dendrobium barbatulum Lindl.

2020 ◽  
Vol 30 (2) ◽  
pp. 243-252
Author(s):  
Ashok N Pyati
Keyword(s):  
Survival Rate ◽  
Plant Regeneration ◽  
Casein Hydrolysate ◽  
Ms Medium ◽  
Organic Additives ◽  
Coconut Husk ◽  
Cane Juice ◽  
Half Strength ◽  
Complex Organic

In vitro regenerative potential of protocorm like bodies (PLBs) of Dendrobium barbatulum were assessed. The formation of secondary PLBs from the primary PLBs that formed from the callus, it is derived from a few larger injured protocorm like bodies. The injured larger PLBs were cultured on half strength MS supplemented with different concentrations of BAP (0.32, 1.62, 3.23 and 16.15 μM), Zn (0.46, 2.28, 4.56 and 22.80 μM) and Kn (0.47, 2.33, 4.65 and 23.25 μM). The highest percentage of secondary PLBs (66.33%) were obtained on half strength MS medium supplemented with BAP 3.23 μM, after 5 weeks of culture. Then these clumps of PLBs were subcultured on half strength MS fortified with coconut water (CW), cane juice (CJ), peptone (P) and casein hydrolysate (CH) for regeneration. Among the complex organic additives 20% CW and 2.0 g/l P to half strength MS resulted in the development of plantlet 78.31 and 62.60%, respectively. Well developed plantlets were successfully acclimatized in the community pots having brick pieces, charcoal, decaying litter and coconut husk (1 : 1 : 1 : 1) gave maximum survival rate of 87.03%. Plant Tissue Cult. & Biotech. 30(2): 243-252, 2020 (December)

scholarly journals In vitro Propagation of Bacopa monnieri (L.)

2020 ◽  
pp. 32-39
Author(s):  
Poornima Raj ◽  
J. Anbumalarmathi ◽  
S. Aruna Sharmili
Keyword(s):  
Survival Rate ◽  
Growth Regulators ◽  
In Vitro Propagation ◽  
Bacopa Monnieri ◽  
Shoot Length ◽  
Ms Medium ◽  
Root Initiation ◽  
Shoot Induction ◽  
Propagation Technique

An experiment was conducted for standardization of in vitro propagation technique of Bacopa monnieri (L.), a medicinal herb of India. Healthy leaf segments of the herb were used as explants with basic Murashige and Skoog (MS) medium containing various combinations of different growth regulators for callus, shoot and root initiation. The best callus induction percentage (95.47%) was observed on MS + 0.5 mg/L NAA and 2.0 mg/L BAP (T3). The maximum number of shoots (8), shoot length (9.30 cm) and shoot induction percentage (90.48%) was achieved on MS + 3.0 mg/L BAP and 1.0 mg/L Kn (ST4). The maximum number of roots (8) and root length (7) was observed on MS + 1.5 mg/L IAA (RT5). The rooted micro shoots were successfully hardened and acclimatized in green house and subsequently established in soil with survival rate of 90%.

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