scholarly journals Kultur Apeks untuk Penyediaan Bibit Unggul Tebu Varietas PS864 dan PS881

2016 ◽  
Vol 10 (2) ◽  
pp. 45
Author(s):  
Deden Sukmadjaja ◽  
Yati Supriyati ◽  
Saptowo J. Pardal
Keyword(s):  
In Vitro Culture ◽  
Genetic Resources ◽  
Genetic Stability ◽  
Shoot Proliferation ◽  
Basic Medium ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Experimental Station ◽  
South Sulawesi

<p>In vitro culture<br />techniques have become alternative to help overcome the<br />problems those are often encountered in the provision of<br />seeds through conventional means. Micropropagation<br />through apex culture in sugarcane has several advantages,<br />such as the produced plants have higher genetic stability,<br />high multiplication rate, and more healthy seeds (especially<br />virus-free)., The aims of the the research were to produce<br />seeds of two varieties of sugarcane, namely PS864 and<br />PS881, through apex culture. Laboratory-scale research was<br />conducted at the Indonesian Center for Agricultural<br />Biotechnology and Genetic Resources Research and<br />Development (ICABIOGRAD), Bogor, while sowing seeds<br />nursery was done in the Experimental Station of Gowa,<br />South Sulawesi Assessment Institute for Agricultural<br />Technology. The experiments consisted of initiation and<br />regeneration of apexes, shoots multiplication, rooting<br />induction, and acclimatization of plantlets. Research results<br />showed the initiation and regeneration of PS864 and PS881<br />through apex culture could be done on MS basic medium<br />containing 0.5 mg/l BAP. Shoot proliferation of both varieties<br />increased in the second subculture. Addition of 1 mg/l BAP<br />into medium in the second subculture resulted in higher<br />average number of shoots than that of 5 mg/l BAP, both for<br />PS864 and PS881. Addition of 1 mg/l and 5 mg/l kinetin<br />showed no significant differences for shoot numbers<br />compared to that of PS864 in medium containing 1 mg/l<br />BAP. The average number of PS881 shoots in multiplication<br />media containing 5 mg/l kinetin was higher than that of 1<br />mg/l kinetin. Increased concentrations of NAA and IBA from<br />0.1 mg/l to 0.5 mg/l in the MS medium were correlated to the<br />increased number of roots in PS864 shoots. Meanwhile, only<br />increased concentration of NAA that affected rooting percentage<br />of PS881. Acclimatization showed the percentage of<br />the plantlets grown in polybags was higher than that directly<br />grown in planting bed. The primary seeds (G0) produced in<br />these experiments were ready to be reproduced again to<br />obtain further stages.</p>

scholarly journals Effects of Cytokinins on in vitro Culture of Bauhinia purpurea L.

2020 ◽  
pp. 161-166
Author(s):  
Belai Meeta Suwal Singh
Keyword(s):  
In Vitro Culture ◽  
Analytical Procedure ◽  
Growth Parameters ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Experimental Purpose ◽  
Bauhinia Purpurea

Bauhinia purpurea L. is a moderate-sized tree with multipurpose value tree yields gum, its bark contains tannin and leaves are used as fodder. It is distributed in sub-Himalayan tracts. The main objective of this study was to evaluate the effects of different cytokinins on growth parameters of B. purpurea and develop a standard micropropagation protocol for nodes and shoot proliferations. The cytokinins used in this study were N-Benzyl-9(2-tetrahydropyranyl) (BPA), 6-furfurylaminopurine, Kinetin (Kn), 6-(4-Hydroxy-3-methyl-trans-2-butenylaminopurine) (Zeatin) (Zin), 2-isopentenyl aminopurine, (2-iP) and 6-benzylaminopurine (BAP) at four different concentrations (0.5, 1.0, 2.0 and 5.0 µM). Murashige and Skoog (1962) (MS) medium was used for the experimental purpose. Multiplication rate of plants was recorded after 8 weeks of culture. Such propagated best grown plants were acclimatized and transferred to the field. All the collected data were worked out statistically with SPSS, a system of analytical procedure

scholarly journals In Vitro Clonal Propagation of Vanilla (Vanilla planifolia ‘Andrews’)

HortScience ◽  
2008 ◽  
Vol 43 (2) ◽  
pp. 454-458 ◽  
Author(s):  
Hilda E. Lee-Espinosa ◽  
Joaquín Murguía-González ◽  
Benjamín García-Rosas ◽  
Ana L. Córdova-Contreras ◽  
Antonio Laguna-Cerda ◽  
...  
Keyword(s):  
Adventitious Shoots ◽  
Shoot Proliferation ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Naphthalene Acetic Acid ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Vanilla Planifolia ◽  
Number Of Shoots

A complete and efficient regeneration protocol was developed for Vanilla planifolia ‘Andrews’, an endangered orchid species that represents an important crop in several tropical countries. Axillary buds excised from the first to the eighth node, considering the first to fourth nodes as “young” (zone 1) and the fifth to eighth as “mature” (zone 2), were cultured on Murashige and Skoog (MS) medium supplemented with 5.73, 7.64, 9.55, or 11.46 μm 6-benzylaminopurine (BA) for shoot induction and in combination with 4.45 μm naphthalene acetic acid (NAA) to induce multiple shoot proliferation. Cytokinin concentration and bud position in the stem had a significant effect on the number of shoots regenerated. The greatest shoot formation per explant, for the two tested zones, was obtained with 9.55 μm BA on MS medium supplemented with 100 mg·L−1 myoinositol, 150 mg·L−1 citric acid, 100 mg·L−1 ascorbic acid, and 20 g·L−1 sucrose. Young buds from zone 1 were able to form an average of 18.5 ± 2.4 shoots per explant, whereas buds from zone 2 induced a maximum of 11.0 ± 1.0 shoots per explant. Plants of 2 to 3 cm height developed a root system in half-strength MS medium supplemented with 0.44 μm NAA and, once they reached 5 cm height on average, were transferred to greenhouse conditions for their acclimatization where a 100% rate survival was achieved. The optimal use of both young and mature buds from each mother plant to induce adventitious shoots permitted a marked increase in the number of shoots per explant. By using all buds from the upper stem part (zone 1 + zone 2) and subculturing every 90 d, the multiplication rate was 1.1 to 1.86 × 105 shoots per bud per year.

scholarly journals In vitro culture of zygotic embryos and seeds of Caesalpinia ferrea Martius

Hoehnea ◽  
2018 ◽  
Vol 45 (4) ◽  
pp. 663-668
Author(s):  
Daniel da Silva ◽  
Angela Maria Imakawa ◽  
Suely de Souza Costa ◽  
Paulo de Tarso Barbosa Sampaio
Keyword(s):  
In Vitro Culture ◽  
Culture Medium ◽  
Germination Rate ◽  
Zygotic Embryos ◽  
Multiplication Rate ◽  
Ms Medium ◽  
In Vitro Germination ◽  
Growth Room ◽  
Morphogenetic Responses

ABSTRACT The aim of this study was to evaluate the in vitro germination of zygotic embryos and seeds of Caesalpinia ferrea Martius and the morphogenetic responses of the explants to different concentrations of growth regulators. Seeds and zygotic embryos were inoculated in MS culture medium and kept in a growth room at a temperature of 25 ± 2 ºC for 16 hours of photoperiod for 30 days. The seeds had a higher in vitro germination rate than the explants from zygotic embryos. However, zygotic embryos in MS medium supplemented with 0.9 mg L-1 BAP had the highest percentage of regeneration (50%), number of shoots (3.25), buds (2.85) and leaves (3.15), multiplication rate (27.75), and length of shoots (1.96 cm). The in vitro culture of zygotic embryos and seeds made possible the multiplication of a higher number of healthy seedlings. Thus, it can be used as an alternative technique for the propagation of this species.

scholarly journals Effects of Sugars on in vitro Culture of Bauhinia purpurea L.

2015 ◽  
Vol 15 (2) ◽  
pp. 47-50
Author(s):  
Belai Meeta Singh
Keyword(s):  
In Vitro Culture ◽  
Science And Technology ◽  
Shoot Length ◽  
The Other ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Bauhinia Purpurea

Bauhinia purpurea L. is a moderate sized tree with multipurpose value. It is distributed in sub-Himalayan tracts. Propagation of this plant was tested on different sugars, maltose, sucrose, glucose, galactose, lactose and fructoseeach with MS medium. Multiplication rate of plants were recorded after 8 weeks of culture. Sucrose produced 2.20 nodes and 10.05 mm shoot length the other sugars did not induce shoot length elongation and node developments.DOI: http://dx.doi.org/njst.v15i2.12113Nepal Journal of Science and Technology Vol. 15, No.2 (2014) 47-50

scholarly journals Development of in vitro culture establishment conditions and micropropagation of grapevine rootstock cultivar ‘Ruggeri-140’

2021 ◽  
Vol 39 ◽  
pp. 03002
Author(s):  
Gayane Melyan ◽  
Andranik Barsegyan ◽  
Narek Sahakyan ◽  
Kima Dangyan ◽  
Yuri Martirosyan
Keyword(s):  
In Vitro Culture ◽  
Shoot Proliferation ◽  
Culture Conditions ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ex Vitro ◽  
Resistant Rootstock

Optimization of in vitro culture conditions of grapevine phylloxera-resistant rootstock cultivar ‘Ruggeri-140’(Vitisberlandieri x Vitisrupestris) was carried out. Among the different sterilization treatments, maximum aseptic cultures were obtained for both explants apical tips and nodal segments when treated with Ca(ClO)2 at concentration of 1.5 % for 10 minutes plus 70 % ethanol for 30 s (T7). The maximum shoot proliferation was observed both in apical and nodal meristems cultured on MS medium supplemented with 1.0 mg/l BAP. MS/2 medium containing 1.0 mg/l indole-3-butric acid (IBA) gave the highest rooting percentage (100%) with the highest mean number and length of roots. The ex vitro survival of rooted micro shoots was 75.0%.

scholarly journals Introduction to in vitro culture and callus initiation in Salvia hispanica L. (chia)

2019 ◽  
Vol 17 (1) ◽  
pp. 33-37
Author(s):  
A. Z. Revutskaya ◽  
A. V. Holubenko ◽  
N. V. Nuzhyna ◽  
H. O. Rudik ◽  
N. Yu. Taran
Keyword(s):  
In Vitro Culture ◽  
Growth Regulators ◽  
Nutrient Medium ◽  
Callus Growth ◽  
Basic Medium ◽  
High Concentration ◽  
Salvia Hispanica ◽  
Callus Initiation ◽  
Salvia Hispanica L

Aim. Preparation of aseptic seedlings Salvia hispanica L., callus initiation in vitro and establishment of primary explants suitable for the callus production. Methods. Seeds are sprouted on our own modification of conventional methods. The non-hormonal Murashige-Skoog agarized nutrient medium was used as basic medium for the experiments. Parts of one-month seedlings (roots, hypocotyl, cotyledon leaves) were used as explants for the use of the colza. We added growth regulators (BAP, 2,4-D) in different concentration combinations into the nutrient medium for callus initiation. Statistical processing was performed in Microsoft Office Excel. Results. Aseptic S. hispanica seedlings have been obtained. The callus growth was initiated on all types of explants, the dependence of the callus intensity on the type of explants and the growth regulators content in the nutrient medium was established. Morphogenic callus and root-regenerants have been obtained. Conclusions. Hypocotyl was the most suitable primary explant for callus growth. Seedlings, leaves and roots showed low morphogenetic capacity. The nutrient medium with an elevated 2,4-D content was the most effective for initiation of callus genesis and proliferation of non-morphogenous callus. A high concentration of 2,4-D in the medium improves S. hispanica callus growth but suppresses its morphogenic ability.Keywords: Salvia hispanica (Chia), in vitro culture, callus.

IN VITRO RESPONSE BY Terminalia arjuna GENOTYPES DURING MICROPROPAGATION

2021 ◽  
Vol 9 (1) ◽  
pp. 44-50
Author(s):  
Meena Choudhary ◽  
◽  
Ashok Gehlot ◽  
Sarita Arya ◽  
Inder Dev Arya ◽  
...  
Keyword(s):  
Shoot Proliferation ◽  
In Vitro Rooting ◽  
Terminalia Arjuna ◽  
Ms Medium ◽  
Demand And Supply ◽  
Half Strength ◽  
Modified Ms Medium ◽  
In Vitro Response ◽  
Different Doses

Terminalia arjuna is an important tree of the medicinal and sericulture industry, commonly known as Arjun. It’s bark is rich in secondary metabolites makes this plant highly valuable in medicine industry to treat cardiovascular disease. Overexploitation due to high demand in medicine, low seed germination, limitations of the conventional method of propagation push this plant towards being endangered. To conserve germplasm of such tree species and meet the requirement in medicinal industry, some non-conventional propagation method like micropropagation has been developed. The present work highlighted the effect of three genotypes (G-1, G-2, and G-3) on tissue culture of T. arjuna situated at Jodhpur, Rajasthan, India. In vitro shoot proliferation was achieved on a modified MS medium enriched with BAP + additives. Among the tested genotypes, Genotype -1 showed maximum bud break response (100%) followed by G-3 (93.33 %) and G-2 (86.66%). Further multiplication of these shoots on modified MS medium containing BAP + NAA + additives gave 11.38±0.26 (G-1), 9.44±0.21 (G-2) and 10.22±0.32 (G-3) shoots. In vitro rooting was done by pulse treatment with IBA for 10 min prior to transfer on hormone free half strength MS medium containing 0.1% activated charcoal. Maximum in vitro rooting was obtained in G-1 (80%) followed by G-3 (71.11%) and G-2 (68.88%). In the present study, it was observed that optimum growth in all three genotypes required different doses of Plant Growth Regulator. Thus, by identifying and multiplying the best performing genotypes the gap between demand and supply of such medicinal plant can be fulfilled.

scholarly journals Optimization of Micropropagation Protocol for Goji Berry (Lycium barbarum L.)

2016 ◽  
Vol 73 (2) ◽  
pp. 141 ◽  
Author(s):  
Alexandru Fira ◽  
Nirmal Joshee ◽  
Victoria Cristea ◽  
Manuela Simu ◽  
Monica Harta ◽  
...  
Keyword(s):  
Shoot Proliferation ◽  
Optimal Number ◽  
Wheat Starch ◽  
Ms Medium ◽  
Ex Vitro ◽  
Benzyl Adenine ◽  
Lycium Barbarum ◽  
Floating Cell ◽  
The Media

Micropropagation of Lycium barbarum cv. 'Ningxia N1' was achieved. The cultures were by initiated by axenical seed germination. The highest shoot proliferation was obtained on the MS media with 1.33 or 2.22 µM benzyl adenine, gelled with wheat starch as an agar alternative. The treatments with 2.22 µM benzyl adenine ensured proliferation rates superior to the ones with 1.33 μM benzyl adenine, but the latter provided longer and more robust shoots. Use of large microcuttings as an explant onto the multiplication media ensured higher in vitro explant survival, higher number of shoots regeneration and more vigorous plantlets. The microcuttings inserted vertically into the media yielded superior growth and multiplication as compared to the microcuttings placed horizontally. The non-rooted, elongated shoots from the treatment 1.33 μM benzyl adenine were either rooted in vitro on a hormone-free MS medium with starch or used for direct ex vitro rooting and acclimatization. The optimal number of microcuttings/vessel for in vitro rooting was 40 and the rooted plantlets were efficiently acclimatized ex vitro by three methods: float hydroculture in floating cell trays, floating perlite, and in Jiffy7 pellets.

In vitro differentiation of theca cells from ovarian cells isolated from postmenopausal women

2020 ◽  
Vol 35 (12) ◽  
pp. 2793-2807
Author(s):  
P Asiabi ◽  
M M Dolmans ◽  
J Ambroise ◽  
A Camboni ◽  
C A Amorim
Keyword(s):  
Growth Factors ◽  
In Vitro Culture ◽  
Protein Detection ◽  
Basic Medium ◽  
Specific Marker ◽  
Mrna Levels ◽  
Theca Cells ◽  
Ovarian Cortex ◽  
Steroid Dehydrogenase

Abstract STUDY QUESTION Can human theca cells (TCs) be differentiated in vitro? SUMMARY ANSWER It is possible to differentiate human TCs in vitro using a medium supplemented with growth factors and hormones. WHAT IS KNOWN ALREADY There are very few studies on the origin of TCs in mammalian ovaries. Precursor TCs have been described in neonatal mice ovaries, which can differentiate into TCs under the influence of factors from oocytes and granulosa cells (GCs). On the other hand, studies in large animal models have reported that stromal cells (SCs) isolated from the cortical ovarian layer can also differentiate into TCs. STUDY DESIGN, SIZE, DURATION After obtaining informed consent, ovarian biopsies were taken from eight menopausal women (53–74 years of age) undergoing laparoscopic surgery for gynecologic disease not related to the ovaries. SCs were isolated from the ovarian cortex and in vitro cultured for 8 days in basic medium (BM) (G1), enriched with growth factors, FSH and LH in plastic (G2) or collagen substrate without (G3) or with (G4) a GC line. PARTICIPANTS/MATERIALS, SETTING, METHODS To confirm TC differentiation, relative mRNA levels for LH receptor (Lhr), steroidogenic acute regulatory protein (Star), cholesterol side-chain cleavage enzyme (Cyp11a1), cytochrome P450 17A1 (Cyp17a1), hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 (Hsd3b1) and hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 2 (Hsd3b2) were assessed. Immunohistochemistry was also performed for their protein detection and a specific marker was identified for TCs (aminopeptidase-N, CD13), as were markers for theca and small luteal cells (dipeptidyl peptidase IV (CD26) and Notch homolog 1, translocation-associated (NOTCH1)). Finally, we analyzed cell ultrastructure before (Day 0) and after in vitro culture (Day 8), and dehydroepiandrosterone (DHEA) and progesterone levels in the medium using transmission electron microscopy (TEM) and ELISA, respectively. MAIN RESULTS AND THE ROLE OF CHANCE Results obtained from qPCR showed a significant increase (P &lt; 0.05) in mRNA levels of Lhr in F2 (floating cells in G2) and G4, Cyp17a1 in G1 and F1 (floating cells in G1) and Hsd3b2 in G1, G2, G3 and G4. Immunohistochemistry confirmed expression of each enzyme involved in the steroidogenic pathway at the protein stage. However, apart from G1, all other groups exhibited a significant (P &lt; 0.05) rise in the number of CD13-positive cells. There was also a significant increase (P &lt; 0.05) in NOTCH1-positive cells in G3 and G4. Ultrastructure analyses by TEM showed a distinct difference between groups and also versus Day 0. A linear trend with time revealed a significant gain (q &lt; 0.001) in DHEA concentrations in the medium during the culture period in G1, G2, G3 and G4. It also demonstrated a statistical increase (q &lt; 0.001) in G2, G3 and G4 groups, but G1 remained the same throughout culture in terms of progesterone levels. LARGE SCALE DATA N/A LIMITATIONS, REASONS FOR CAUTION Shorter periods of in vitro culture (e.g. 2, 4 and 6 days) could have led to increased concentrations of differentiated TCs in G2, G3 and G4. In addition, a group of cells cultured in BM and accompanied by COV434 cells would be necessary to understand their role in the differentiation process. Finally, while our results demonstrate that TCs can be differentiated in vitro from cells isolated from the cortical layer of postmenopausal ovaries, we do not know if these cells are differentiated from a subpopulation of precursor TCs present in ovarian cortex or ovarian SCs in general. It is therefore necessary to identify specific markers for precursor TCs in human ovaries to understand the origin of these cells. WIDER IMPLICATIONS OF THE FINDINGS This is a promising step toward understanding TC ontogenesis in the human ovary. Moreover, in vitro-generated human TCs can be used for studies on drug screening, as well as to understand TC-associated pathologies, such as androgen-secreting tumors and polycystic ovary syndrome. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (FNRS) (C.A.A. is an FRS-FNRS Research Associate; grant MIS #F4535 16 awarded to C.A.A.; grant 5/4/150/5 awarded to M.M.D.; grant ASP-RE314 awarded to P.A.) and Foundation Against Cancer (grant 2018-042 awarded to A.C.). The authors declare no competing interests.

scholarly journals Micropropagation and in vitro conservation of Alcantarea nahoumii (Bromeliaceae), an endemic and endangered species of the Brazilian Atlantic Forest

2020 ◽  
Vol 42 ◽  
pp. e52940
Author(s):  
Simone Sacramento dos Santos Silva ◽  
Everton Hilo de Souza ◽  
Fernanda Vidigal Duarte Souza ◽  
Cristina Ferreira Nepomuceno ◽  
Maria Angélica Pereira de Carvalho Costa
Keyword(s):  
Atlantic Forest ◽  
Culture Media ◽  
In Vitro Conservation ◽  
Naphthaleneacetic Acid ◽  
High Survival ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Mature Seeds ◽  
Stem Segments

Alcantarea nahoumii (Leme) J. R. Grant is a species native to the Atlantic Forest that stands out for ornamental purposes. The objective of this work was to evaluate the in vitro germination of A. nahoumii seeds and establish a micropropagation protocol for production of seedlings so as to minimize the effects of predatory extractivism and develop an in vitro conservation system. Mature seeds were disinfested, established in three culture media (MS, MS½ and MS⅓) and incubated at four temperatures (20, 25, 30 and 35ºC) in a germination chamber. In the micropropagation experiment, stem segments were introduced in MS medium supplemented with 0.5 μM of 1-naphthaleneacetic acid (NAA) and 0.0, 2.2, 4.4 and 6.6 μM of 6-benzylaminopurine (BAP). For the in vitro conservation, plantlets were established in MS or MS½ medium supplemented with 15 g L-1 or 30 g L-1 of sucrose. The plants were acclimated with commercial substrate. The highest seed germination percentages were promoted by temperature conditions of 20 and 25ºC, with MS culture medium. The highest multiplication rate of shoots was obtained from the treatment without addition of the growth regulator or when combined with 2.2 μM of BAP + 0.5 μM of NAA. The acclimation of the plants occurred with high survival rate. The species can be conserved in vitro under slow growth condition for 24 months when incubated in MS medium supplemented with 30 g L-1 of sucrose.

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