scholarly journals Growth Evaluation of In-Vitro Propagated Embryo of Morinda Citrifolia L. Seeds

2021 ◽  
Vol 25 (1) ◽  
pp. 109-111
Author(s):  
J.O. Afolabi ◽  
D.A. Adegboyega ◽  
Y.O. Fasakin
Keyword(s):  
Embryo Culture ◽  
Basal Medium ◽  
Shoot Development ◽  
Morinda Citrifolia ◽  
Culture Initiation ◽  
The Media ◽  
Number Of Leaves ◽  
Growth Evaluation ◽  
Benzyl Amino Purine

The dormant nature of Morinda citrifolia seeds is a limitation to its efficient in-vitro plantlet multiplication. Hence, the use of embryo culture for successful in-vitro culture initiation. Matured embryo of freshly collected noni seeds were cultured on Murashige and Skoog basal medium supplemented with kinetin (Kn) and Benzyl amino purine (BAP) in the range of A: control (no addition); B: 0.5 mg/l Kn+1.0 mg/l BAP; C: 1.0 mg/l Kn+2.0 mg/l Bap; D: 1.5 mg/l Kn+3.0 mg/l BAP and E: 2.0 mg/l Kn+4.0 mg/l BAP. The results at 4 weeks after inoculation (WAI) showed that germination was faster from medium A without hormone whereas highest percentage germination was obtained from both medium D and E with 80 %. Medium B and C had 65 % each while medium A gave the least (40%). The development of the plantlets showed that longest shoot (3.9 cm) from medium A was closely related to 3.58 cm from Medium B while root lengths (2.28 cm) and number of adventitious roots (26) from medium A were significantly higher than other media at 12 WAI. Highest number of nodes (2.25) obtained from medium D was comparable to Media C and B while medium A had the least at 12 WAI. Number of leaves obtained was similar between the media at 12 WAI. These results indicated that using embryo is reliable for fast in-vitro propagation and shoot development of noni plant with optimum cytokinins (0.5/1.0 mg/l Kn/BAP) application. Keywords: Culture initiation, Cytokinins, Embryo culture, Plantlet, Shoot development

scholarly journals In vitro Propagation of Noni (Morinda citrifolia L.) Through Embryo Culture

2020 ◽  
Vol 30 (2) ◽  
pp. 199-207
Author(s):  
JO Afolabi ◽  
EO Oloyede ◽  
AO Akala
Keyword(s):  
Embryo Culture ◽  
In Vitro Propagation ◽  
Zygotic Embryo ◽  
Shoot Length ◽  
Morinda Citrifolia ◽  
Distilled Water ◽  
Number Of Leaves ◽  
And Control ◽  
Mature Zygotic Embryo

The chances of shoot-regeneration from embryo culture of Morinda citrifolia L. seeds was investigated. Germination on different strengths of MS and control (Sterile distilled water) started by two weeks after inoculation (WAI). At 6 WAI, 90% of the embryo had germinated from 25% MS followed by 80% in control, 70% from 50% MS and 40% each from 100 and 75% MS. Similarly, the same MS media strengths with basal application of 2.0/1.0 mg/l BAP/Kn affected the growth of regenerated Noni-plantlets. The longest shoot length (3.46 cm) and the number of nodes (1.75) were obtained from 75% MS while the highest number of leaves (7.25) was obtained in 100% MS between 4 and 12 WAI. The lowest value for these parameters were observed in 25% MS. This showed that mature zygotic embryo is good explant for the establishment of highly viable and re-generable plantlets of Noni. Plant Tissue Cult. & Biotech. 30(2): 199-207, 2020 (December)

scholarly journals Production of bergenin, an active chemical constituent in the callus of Bergenia ciliata (Haw.) Sternb.

2012 ◽  
Vol 8 ◽  
pp. 40-44 ◽  
Author(s):  
Umesh Krishna Shrestha ◽  
Bijaya Pant
Keyword(s):  
Leaf Explants ◽  
Chemical Constituent ◽  
Wild Plants ◽  
Plant Science ◽  
Naphthalene Acetic Acid ◽  
Active Chemical ◽  
In The Wild ◽  
The Media ◽  
Benzyl Amino Purine

In vitro culture of Bergenia ciliata (Haw.) Sternb. was carried out for the examination of bergenin content. Leaf explants were cultured in MS (Murashige and Skoog) basal media supplemented with or without phytohormones. The hormonal series maintained were in the range of 0-2 mg l-1 for BAP (6-benzyl amino purine) and 0-1.5 mg l-1 for NAA (α-naphthalene acetic acid). Bergenin content of in vitro grown tissues of B. ciliata was compared with that of wild plants collected from three different localities of Nepal. The best growth of callus and plantlets occurred in the media containing BAP 1.0 mg l-1 + NAA 1.0 mg l-1 and BAP 1.5 mg l-1 + NAA 1.0 mg l-1. Production of bergenin was high in the media supplemented with 1.0 mg l-1 BAP + 1.5 mg l-1 NAA (3.40 μg g-1) and 2.0 mg l-1 BAP + 1.5 mg l-1 NAA (3.05 μg g-1) under experimental condition. The bergenin content in the wild plants collected from Langtang, Jumla and Godawari was found to be 4.28 μg g-1, 4.53 μg g-1 and 3.64 μg g-1 respectively. This study shows that the in vitro cultured callus of B. ciliata is capable of synthesizing bergenin in quantity comparable to that of the wild plant.doi: http://dx.doi.org/10.3126/botor.v8i0.5557 Botanica Orientalis – Journal of Plant Science (2011) 8: 40-44

scholarly journals PENGARUH BAP (Benzyl Amino Purine) DAN AIR KELAPA TERHADAP PERTUMBUHAN TUNAS PUCUK DAN KANDUNGAN SULFORAFAN BROKOLI (Brassica oleracea L. var. italica Plenck) SECARA IN-VITRO

2018 ◽  
Vol 14 (1) ◽  
pp. 439
Author(s):  
Alfrida ., Maninggolang ◽  
Jeany Sh. Polii-Mandang ◽  
Wenny ., Tilaar
Keyword(s):  
Brassica Oleracea ◽  
Coconut Water ◽  
Shoot Bud ◽  
Randomized Design ◽  
Wet Weight ◽  
Number Of Leaves ◽  
Brassica Oleracea L ◽  
Benzyl Amino Purine ◽  
Number Of Shoots

This study aims to know the effect of Benzyl Amino Purine (BAP) and Coconut Water on shoot bud growth and Broccoli Sulforaphane content (Brassica oleracea L. var italic Plenck). The study was conducted in the laboratory of Biotechnology Department of Aquaculture, Faculty of Agriculture of Sam Ratulangi University, Manado, that conducted from August-December 2017. This study used a Complete Randomized Design (RAL), consisting of 8 treatments and each repeated as many 4 times, so we get 32 unit experiment. The variables observed were number of buds, number of leaves, plant height, wet weight, root number and Sulforaphane content analysis. The result of research shows that analysis of variance showed that in the use of Benzyl Amino Purine (BAP) concentration 3 ppm tends to increase the number of leaves aged 4 Weeks After Culture (MSK) and increase the number of shoots age 2 and 6 Weeks After Culture (MSK). Benzyl Amino Purine (BAP) 3 ppm can increase the wet weight of age 6W eeks After Culture ((MSK). Coconut water 20% tends to increase the number of leaves at age 6 Weeks After Culture (MSK) and increase the number of shoots aged 6 Weeks After Culture (MSK), while for combination of 3 ppm Benzyl Amino Purine (BAP) and coconut water 20% tends to increase the number of leaves aged 2 Weeks After Culture (MSK) and the number of shoots aged 2 Weeks After Culture (MSK). Combination of coconut water and Benzyl Amino Purine (BAP) is not detected by the content of Sulforaphane.

scholarly journals Mass Propagation of Nauclea diderrichii (de Willd. & Th. Dur) Merr. Seedlings through Tissue Culture Technique

2021 ◽  
Vol 31 (1) ◽  
pp. 51-60
Author(s):  
RI Oyediran ◽  
JO Afolabi ◽  
DB Olomola ◽  
FO Akanni
Keyword(s):  
Tissue Culture ◽  
Adventitious Shoots ◽  
Basal Medium ◽  
Culture Technique ◽  
Seedling Production ◽  
Mass Propagation ◽  
In Vitro Plantlets ◽  
Number Of Leaves ◽  
Nauclea Diderrichii

Nauclea diderrichii is a tree species of economic importance. However, its plantation establishment is limited by inadequate seedling production. Hence, there is ample scope of tissue culture for its mass propagation. Its in vitro plantlets development as affected by media strengths indicated that 100 % seed germination was obtained in full MS basal medium while the least (3.35 %) was from quarter-strength at 8 Weeks after inoculation (WAI). The effects of BAP and NAA assessed on the growth of its sub-cultured plantlets showed that highest number of leaves (17) and adventitious shoots (3) were obtained from MS basal medium supplemented with 0.1 mg/l BAP only. Whereas, highest shoot length (3.61 cm) and average number of roots (5/plantlet) were obtained from the same medium without hormone(s) at 8 WAI. Further sub-culturing into MS with 0.05 mg/l NAA resulted into plantlets having optimum shoot and massive root growth ready for acclimatization in 6 WAI. The plantlets were successfully acclimatized using coconuthusk/ topsoil mixture with 90 % survival. Plant Tissue Cult. & Biotech. 31(1): 51-60, 2021 (June)

scholarly journals 319IN VITRO MATURATION OF EQUINE OOCYTES IN A COMPLETELY DEFINED MEDIUM SUPPLEMENTED WITH PROGESTERONE

2004 ◽  
Vol 16 (2) ◽  
pp. 279
Author(s):  
B. Merlo ◽  
E. Iacono ◽  
F. Prati ◽  
G. Mari
Keyword(s):  
Polar Body ◽  
Basal Medium ◽  
Defined Medium ◽  
Chi Square ◽  
Nuclear Maturation ◽  
Maturation Rate ◽  
Chi Square Test ◽  
The Media ◽  
Cytoplasmic Maturation

A completely defined medium for in vitro maturation (IVM) of equine oocytes has not yet been developed, since most of the media used for IVM are supplemented with serum or BSA. Furthermore, in this species there is no report about the influence of progesterone on maturation, although it has already been used as supplement (500ngmL−1) in EMMI (Maclellan LJ et al., 2001, Theriogenolgy 55, 310 abst). The aims of this study were to develop a completely defined medium for equine oocyte maturation and to investigate the effect of progesterone on nuclear maturation. Equine oocytes were collected by follicular scraping of abattoir-derived ovaries between April and June. The basal medium for maturation was SOFaa supplemented with pFSH-LH 0.1IUmL−1 (Pluset, Laboratorios Calier, Barcelona, Spain), EGF* 50ngmL−1, ITS (Insulin, Transferrin, Sodium selenite), L-cysteine 1.2mM, Maturation SOF (MSOF). Compact cumulus-oocyte complexes were selected, washed three times in H-SOF and matured in one of the following media (15–20 oocytesmL−1): (1) MSOF+FCS 10% (MSOF-FCS), (2) MSOF+progesterone 100ngmL−1 (MSOF-P4), (3) MSOF. After 24h of culture in 5% CO2 in air at 38.5°C, the oocytes were denuded by gently pipetting in a 0.25% trypsin solution, washed and stained with Hoechst 33258 (10μgmL−1 in PBS) for 30min at room temperature. Oocytes were examined under a fluorescent microscope to assess nuclear maturation. Only oocytes with an evident polar body and metaphase II plate (MII) were considered mature. The experiment was done in 6 replicates. Chi Square test was used for statistical analysis (Statistica for Windows – Stat Soft Inc., Tusla, OK, USA). Significance was assessed for P<0.05. The results of this study show that MSOF can be considered a suitable completely defined medium for IVM of equine oocytes. Adding progesterone significantly (P<0.05) increases the nuclear maturation rate at 24h of culture. It can be speculated that although cumuls cells produce this hormone, supplementation is useful to reach progesterone concentrations similar to those present in follicular fluid (early dominant 63.4±19.3ngmL−1, healthy preovulatory follicle 1094.3±170.9ngmL−1; Gerard N et al., 2002, Reproduction 124, 241–248). Further studies are needed to investigate the influence of progesterone on cytoplasmic maturation and to test the effect of different progesterone concentrations and time of maturation in a completely defined system.*All chemicals were purchased from Sigma, St. Louis, MO, USA, unless otherwise stated. Table 1 Maturation of equine oocytes in different media

120 EFFECT OF INDIVIDUAL CULTURE SYSTEM USING WOW OR CELL-TAK ON DEVELOPMENT OF IN VITRO-PRODUCED BOVINE EMBRYOS

2009 ◽  
Vol 21 (1) ◽  
pp. 160
Author(s):  
S. Matoba ◽  
P. Lonergan
Keyword(s):  
Embryo Culture ◽  
Culture System ◽  
Basal Medium ◽  
Bovine Embryos ◽  
Paracrine Effects ◽  
Individual Culture ◽  
Cell Adhesive ◽  
Developmental Rates ◽  
One Way Anova

The culture of embryos individually in vitro is generally associated with poorer developmental rates. However, the ability to do this successfully would greatly facilitate studies where identification of individual embryos, or the embryos from a particular donor, is necessary. The objective of this study was to examine the effect of culture system on the development of individual IVP bovine embryos. Presumptive zygotes (n = 1301, 6 replicates), produced by IVM/IVF, were used. The aim of Experiment 1 was to compare development of bovine embryos in SOF or CR1aa supplemented with 5% FCS. Zygotes were cultured in droplets under oil as follows: (i) 20/25 μL, (ii) 20/100 μL or (iii) 20/100 μL individually in the Well of the Well (WOW) system (Vajta et al. 2000 Mol. Reprod. Dev. 55, 254–264). Twenty WOW were prepared in a 100 μL droplet of medium under oil using a sterile rod. The aim of Experiment 2 was to compare development of embryos cultured in groups but individually identifiable on the cell adhesive Cell-Tak (Stokes et al. 2005 Dev. Biol. 284, 62–71) or in the WOW system. Zygotes were cultured as follows: (i) 20/20 μL, (ii) 20/20 μL with Cell-Tak, (iii) 20/100 μL with Cell-Tak or (iv) 20/100 μL in WOW. A drop of Cell-Tak (1 μL/20 μL medium) was placed on the base of the dish, dried for 20 min, washed with sterile water and dried completely. Once dried, the area was covered with 10 μL of FCS-free medium and groups of 20 zygotes were placed on the Cell-Tak in a 5 × 4 grid formation a maximum of 160 μm apart. Then, an additional 10 μL or 90 μL medium supplemented with FCS was added to give a final volume of 20 or 100 μL. Cleavage and blastocyst rates were assessed on Day 2 and Days 7–9, respectively. Data (means ± SE) were analyzed by one way ANOVA. In Experiment 1, there were no differences between SOF and CR1aa with respect to culture of embryos individually in WOW (P > 0.05); therefore, SOF was used as the basal medium for Experiment 2. There were no differences (P > 0.05) between the cleavage and blastocyst rate among drop sizes and individual culture systems; individual culture, irrespective of the system used (Cell-Tak or WOW), resulted in the similar developmental rates to the control. In conclusion, individual embryo culture offers the opportunity to study embryo development in a more powerful manner. Furthermore, the use of the cell adhesive Cell-Tak may be more practical because it removes the potential variability associated with well dimensions in the WOW system and may improve any potential paracrine effects during embryo culture. Further studies are required to establish the viability of such embryos after transfer. Table 1.Effect of individual culture system on development of IVP bovine embryos Supported by Science Foundation Ireland.

scholarly journals In vitro meristem-tip culture and regeneration approaches in Congolese cassava accessions (Manihot esculenta Crantz cv. Boma and cv. Mpelo-Nlongi)

2015 ◽  
Vol 3 (2) ◽  
pp. 72
Author(s):  
J.-Roger Bansimba Mukiese ◽  
Aimé Diamuini Ndofunsu ◽  
Freddy Bulubulu ◽  
Alexandre Mbaya Ntumbula ◽  
Sébastien Luyindula Ndiku
Keyword(s):  
Growth Regulators ◽  
Manihot Esculenta ◽  
Callus Formation ◽  
Shoot Development ◽  
Media Composition ◽  
Manihot Esculenta Crantz ◽  
Adenine Sulphate ◽  
Meristem Tip Culture ◽  
The Media

<p>Shiny dome-like structures measuring less than 1mm in length were excised aseptically from shoot tip buds of infected of two cassava (<em>Manihot esculenta</em> Crantz) local cultivars (Boma and Mpelo Nlongi) and cultivated <em>in vitro</em> in two types of media with different combination of growth hormone: Murashige and Skoog supplemented of sucrose (20 g/l), Naphtalenacetic acid (NAA, 10 μM), Ben-zylaminopurine (BAP, 0.66 μM) as well as Gibberellic acid (GA3, 0.1 μM) with 80 mg/l of Adenine sulphate and MS-free growth regulators. After four weeks, data were scored: 29.5% responding explant with callus formation and 20.5% responding explants to shoot development in the medium with growth regulators for the cultivar Boma whereas the cultivar Mpelo-Nlongi presented 5.7% and 25.7% respectively of callus formation and shoot development. The cultivar Boma presented a tendency more pronounced for the callus formation rather than with the shoot development contrary to the cultivar Mpelo-Nlongi. In regards of this experiment, it was shown that the media composition and genotype are essential factors, which influence in vitro growth, mainly the shoot development, in the culture of meristems for cassava local accessions.</p>

scholarly journals Response of in vitro propagated fig (Ficus carica L.) shoots to the concentrations of benzyl amino purine and coconut water

2021 ◽  
Vol 922 (1) ◽  
pp. 012067
Author(s):  
Sophia ◽  
M Hayati ◽  
E Kesumawati
Keyword(s):  
Callus Formation ◽  
Formation Rate ◽  
Water Concentration ◽  
Shoot Formation ◽  
Coconut Water ◽  
Contamination Rate ◽  
Two Factors ◽  
Number Of Leaves ◽  
Benzyl Amino Purine

Abstract In this study, several concentrations of benzyl amino purine (BAP) and coconut water (CW) were investigated along with the interaction between two factors to the growth of in vitro propagated fig shoots. The investigated factors consisted of BAP concentration: 0, 1, 3, 5 mg L−1 and coconut water concentration: 0, 100, 200, 300 ml L−1. A total of 16 treatment combinations with 6 replications resulting in 96 experimental units consisting of a single fig shoot explant per culture medium. The observed parameters including living explant rate, contamination rate, browning rate, day of first shoot emergence, shoot formation rate, explant height addition, number of leaves, callus formation rate, and number of roots were conducted every week from 1 to 8 weeks after proliferation (WAP). The result indicated that in 8 WAP, the living explant rate reached 23.95%. The combination of concentration 200 ml L−1 CW and 3 mg L−1 BAP + 200 ml L−1 CW-induced early emergence of new shoots at 7 days after proliferation (DAP). The highest shoot formation rate (100%) was observed at a concentration of 300 mL L−1CW. The highest explant height addition (7.10 cm) was observed at a concentration of 200 mL L−1 CW. The highest number of leaves (5.80) was observed at a concentration of 1 mg L−1 BAP + 200 mL L−1 CW. The highest callus formation rate (50%) was observed at a concentration of 100 ml L−1CW and 300 ml L−1 CW. The highest number of roots (17) was observed in the control.

scholarly journals Kajian Penambahan BahanOrganik Pada Media Tanam VW Pada Organogenesis AnggrekDendrobium SecaraIn Vitro

2018 ◽  
Vol 1 (1) ◽  
Author(s):  
Siti Rahmah ◽  
Tintrim Rahayu ◽  
Ari Hayati
Keyword(s):  
Tissue Culture ◽  
Organic Matter ◽  
Optimization Technique ◽  
Media Composition ◽  
Growth Optimization ◽  
The World ◽  
The Media ◽  
Number Of Leaves ◽  
Bean Sprouts

In vitro tissue culture is a growth optimization technique of Dendrobium orchid with according to media composition. Nutritions in the media are important for dendrobium orchid. Dendrobium orchid  include plant from orchidaceae family its spread throughout the world like indonesia. Its features are easily planted, intersest is continuous and varied, easily assembled, the flower crown is not easy to fall and wither. Research aimed at obtaining media compositions that are easily available and able to fulfill the needs of orchid plants. The research was conducted using descriptive methods to compare different trearment; Vacin & Went and VW media with adding organic matter; extract bean sprouts, potato extrac, and water coconut; wich is conducted for eight weeks after planting. The result of addition organic matter on VW media was different toward organogenesis of orchid. The average number of shoots is 1.8; the number of leaves average of 6.8 and the number of roots average of 3.6 formed from two until eight weeks after culture.Keywords: tissue culture, growing media, Dendrobium orchid, organogenesis.ABSTRAKKultur jaringan in vitro adalah salah satu teknik optimalisasi pada pertumbuhan tanaman angrek Dendrobium dengan menyesuaikan komposisi medianya. Nutrisi yang terdapat di dalam media sangat penting bagi pertumbuhan anggrek. Anggrek Dendrobium termasuk tanaman dari keluarga Orchidaceae yang penyebarannya sampai ke pelosok dunia seperti Indonesia. Keistimewaanya mudah ditanam, bunganya terus-menerus dan bermacam-macam, mudah disusun, serta mahkota bunga tidak mudah jatuh dan layu. Penelitian yang bertujuan untuk mendapatkan komposisi media yang mudah didapat dan mampu memenuhi kebutuhan tanaman anggrek. Penelitian dilakukan menggunakan metode deskriptif untuk membandingkan perlakuan media yang berbeda yaitu media Vacin & Went, dan VW dengan penambahan bahan organik; ekstrak tauge kacang hijau, ekstrak kentang, dan air kelapa muda; yang dilakukan selama delapan minggu setelah tanam. Hasil penambahan bahan organik pada media VW berbeda terhadap organogenesis eksplan anggrek. Jumlah tunas rata-rata 1,8; Jumlah daun rata-rata 6,8 dan jumlah akar rata-rata 3,6 yang terbentuk dari dua minggu setelah kultur (MSK) sampai minggu terakhir pengamatan delapan MSK.Kata kunci: kultur jaringan, media tanam, angrek Dendrobium,organogenesis.

Ovule and Embryo Culture of Arachis hypogaea and Interspecific Hybrids1

1988 ◽  
Vol 15 (2) ◽  
pp. 98-104 ◽  
Author(s):  
H. T. Stalker ◽  
M. A. Eweda
Keyword(s):  
Embryo Culture ◽  
Interspecific Hybrids ◽  
Shoot Growth ◽  
Basal Medium ◽  
Lower Percentage ◽  
Naphthaleneacetic Acid ◽  
Wide Crosses ◽  
In Vitro Techniques ◽  
Basal Media

Abstract Interspecific hybridization in Arachis is difficult between species within sectional groups and nearly impossible among more distantly related species. Embryos usually abort early in the reproductive cycle; thus in vitro techniques are necessary to recover many desirable hybrid combinations in the genus. The objectives of this investigation were to develop techniques whereby mature plants could be recovered from otherwise aborting embryos. First, ovule culture was performed using eight genotypes, three levels of kinetin, and the two basal media Murashige and Skoog (MS) and N6. Two-tenths mg/L kinetin in media resulted in 24% of the ovules swelling to a size of 3-4 mm which could be used for excising embryos. Embryo culture was next performed on five genotypes. The transfer series (I) 0.2 mg/L kinetin for 21 days, (2) 0.5 mg/L 6-benzylamino-purine (BAP) for 14 days and, (3) MS without growth regulators resulted in 34.6% of ovules producing plants across genotypes; other transfer series either resulted in a lower percentage of plant recovery and/or tissues of some genotypes which did not survive to maturity. The BAP medium induced shoot growth, while root growth was induced on the MS without growth regulator medium. Approximately 90% of embryos transferred to a mist system after 7-9 weeks in vitro survived transplanting to soil. Two interspecific hybrids were recovered from incompatible hexaploid x diploid crosses, but only after roots were induced using a MS basal medium with 4 mg/L 1-naphthaleneacetic acid:2 mg/L indole-3-butyric acid in a fourth tissue transfer. The experiments illustrated the feasibility of rescuing embryos of A. hypogaea and interspecific peanut hybrids. The process is slow and will be most applicable to wide crosses which cannot be obtained by more conventional methods.

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