scholarly journals Perbanyakan Massal Embrio Kalamondin Melalui Teknologi Somatik Embriogenesis Menggunakan Bioreaktor

2013 ◽  
Vol 22 (1) ◽  
pp. 1
Author(s):  
Nirmala Frianti Devy ◽  
Farida Yulianti ◽  
Hardiyanto Hardiyanto
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Multiplication Rate ◽  
Mass Propagation ◽  
Liquid Media ◽  
Embryogenic Calli ◽  
Research Results ◽  
The Real ◽  
Subtropical Fruit

<p>Sejauh ini, penelitian perbanyakan somatik embriogenesis baik untuk penyediaan semaian batang bawah maupun varietas komersial jeruk menghasilkan laju multiplikasi yang relatif lambat. Kombinasi antara perbanyakan melalui metode somatik embriogenesis dengan penggunaan bioreaktor, diharapkan mampu meningkatkan laju produksi kalus embrionik menjadi planlet.  Kajian awal dilakukan menggunakan nuselus Kalamondin (Citrus mitis Blanco) sebagai sumber kalus. Kalus yang dihasilkan diinduksi dan diperbanyak menjadi kalus embrionik dan embrio dengan cara dikulturkan pada shaker (100 rpm) serta bulb bioreactor. Tujuan penelitian ini ialah membandingkan produksi embrio Kalamondin melalui teknologi somatik embriogenesis pada kultur cair menggunakan shaker dan bioreaktor. Penelitian dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Jeruk dan Buah Subtropika, dari September 2008 sampai dengan Desember 2009.  Pada tahapan perbanyakan embrio dengan metode shaker, diperoleh bahwa rerata kemampuan kalus menghasilkan embrio dalam kultur selama 10 minggu ialah 18,12 embrio/g kalus. Dengan kisaran waktu yang sama, total embrio yang dihasilkan 3 g kalus/300 cc media cair di dalam bioreaktor menghasilkan 46 embrio/g kalus atau setara 2,53 kali dibandingkan metode shaker. Embrio yang tumbuh pada bioreaktor dapat berkembang hampir 100% menjadi planlet. Hasil penelitian ini membuktikan bahwa aplikasi bioreaktor untuk tujuan perbanyakan massal embrio Kalamondin memiliki pengaruh yang signifikan terhadap laju multiplikasinya.</p><p>ABSTRAK</p><p>So far, research on somatic embryogenesis for rootstock and citrus commercial varieties has been faced by low multiplication rate of embryos. Combination of somatic embryogenesis method and bioreactor hypothezed can increase multiplication rate of embryos and improve regeneration of embryogenic calli to produce plantlets.  Kalamondin explants were inducted and proliferated to be embryonic calli and embryos using both shaker (100 rpm) and bulb bioreactor. The aimed of this research was to compare the production of Kalamondin embryos through somatic embryogenesis method on liquid media using shaker and bulb bioreactor. Research was conducted at Tissue Culture Laboratory of Indonesian Citrus and Subtropical Fruit Research Institute from September 2008 to December 2009.  Kalamondin nucelus as a callus source was used in this research. Results of the study indicated that the average of embryos production through shaker technique within 10 weeks of culture incubation was 18.12 embryos/g callus, while application of bioreactor imrpoved embryo productivity up to  46 embryos/g calli (3 g/300 cc media). The multiplication rate using the bioreactor increased up to 2.53 fold compare to shaker method. Results of the study give the real evidence that application of biorector for in vitro mass propagation of Kalamondin embryos had high significant effect on embryo multiplication rate.</p>

scholarly journals Plant Regeneration of Allium victorialis var. platyphyllum Makino via Organogenesis and Somatic Embryogenesis

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 628a-628
Author(s):  
Hak-Tae Lim ◽  
Eun-Ae Lee ◽  
Won-Bae Kim
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Multiple Shoots ◽  
Root Induction ◽  
Ms Medium ◽  
Mass Propagation ◽  
Embryogenic Calli ◽  
Wild Vegetable ◽  
Shoot Tip Explants

This study was conducted to investigate the possibility of obtaining plantlets via somatic embryogenesis and organogenesis as means of in vitro mass propagation in Allium victorialis var. platyphyllum Makino, one of the most popular wild vegetable plants in Korea. Shoots formed directly when bulb explants of A. victorialis were cultured on MS medium containing 0.2 mg·L–1 NAA and 2.0 mg·L–1 zeatin under 16 hours (light)/8 hours (dark) illumination. The use of leaf and shoot tip explants was not successful, largely due to explant senescence in the present of plant growth regulators. Embryogenic calli were obtained from the bulb explants of A. victorialis on MS medium supplemented with 0.2 mg·L–1 NAA, 0.2 mg·L–1 BAP, and 1.0 mg·L–1 picloram after 4–5 weeks of culture in the dark at 27°C. Upon transfer to shoot-induced MS medium containing 0.2 mg·L–1 NAA and 2.0 mg·L–1 zeatin, embryogenic calli gave rise to numerous somatic embryos, which subsequently developed into multiple shoots after 3 months of culture under 16 hours(light)/8 hours (dark) illumination. For root induction, regenerated shoots were transferred to MS medium added with 1.0 mg·L–1 NAA. Regenerants with well-developed roots were potted in an artificial soil mixture of vermiculite (1) and perlite (1).

scholarly journals Barley somatic embryogenesis-an attempt to modify variation induced in tissue culture

2021 ◽  
Vol 28 (1) ◽  
Author(s):  
Renata Orłowska
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Taguchi Method ◽  
Culture Conditions ◽  
Dna Demethylation ◽  
Silver Ion ◽  
Donor Plant ◽  
In Vitro Tissue Culture ◽  
Induced Variation

Abstract Background Somatic embryogenesis is a phenomenon carried out in an environment that generates abiotic stress. Thus, regenerants may differ from the source of explants at the morphological, genetic, and epigenetic levels. The DNA changes may be the outcome of induction media ingredients (i.e., copper and silver ions) and their concentrations and time of in vitro cultures. Results This study optimised the level of copper and silver ion concentration in culture media parallel with the induction medium longevity step towards obtaining barley regenerants via somatic embryogenesis with a minimum or maximum level of tissue culture-induced differences between the donor plant and its regenerants. The optimisation process is based on tissue culture-induced variation evaluated via the metAFLP approach for regenerants derived under varying in vitro tissue culture conditions and exploited by the Taguchi method. In the optimisation and verification experiments, various copper and silver ion concentrations and the different number of days differentiated the tested trials concerning the tissue culture-induced variation level, DNA demethylation, and de novo methylation, including symmetric (CG, CHG) and asymmetric (CHH) DNA sequence contexts. Verification of optimised conditions towards obtaining regenerants with minimum and maximum variability compared to donor plants proved useful. The main changes that discriminate optimised conditions belonged to DNA demethylation events with particular stress on CHG context. Conclusions The combination of tissue culture-induced variation evaluated for eight experimental trials and implementation of the Taguchi method allowed the optimisation of the in vitro tissue culture conditions towards the minimum and maximum differences between a source of tissue explants (donor plant) and its regenerants from somatic embryos. The tissue culture-induced variation characteristic is mostly affected by demethylation with preferences towards CHG sequence context.

scholarly journals EFFECT OF DIFFERENT SOURCES OF PLANT GROWTH REGULATOR ON THE INDUCTION AND DEVELOPMENT OF MANGOSTEEN SOMATIC EMBRYOS

2017 ◽  
Vol 17 (1) ◽  
pp. 9
Author(s):  
Yosi Zendra Joni ◽  
Riry Prihatini ◽  
Darda Efendi ◽  
Ika Roostika
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Growth ◽  
Embryogenic Callus ◽  
Plant Growth Regulator ◽  
Somatic Embryos ◽  
Casein Hydrolysate ◽  
Malt Extract ◽  
Mass Propagation ◽  
Embryogenic Callus Induction

<p>Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regu-lators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l-1) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l-1) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l-1. The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l-1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l-1 TDZ and 0.7 mg l-1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.</p>

scholarly journals In vitro somatic embryogenesis of superior clones of robusta coffee from Lampung, Indonesia: Effect of genotypes and callus induction media

2020 ◽  
Vol 21 (8) ◽  
Author(s):  
Dwi Hapsoro ◽  
Rahmadyah Hamiranti ◽  
Yusnita Yusnita
Keyword(s):  
Somatic Embryogenesis ◽  
Callus Induction ◽  
Leaf Explants ◽  
Regeneration Medium ◽  
Embryogenic Calli ◽  
Robusta Coffee ◽  
Primary Callus ◽  
Ascorbic Acids ◽  
Superior Clones

Abstract. Hapsoro D, Hamiranti R, Yusnita Y. 2020. In vitro somatic embryogenesis of superior clones of robusta coffee from Lampung, Indonesia: Effect of genotypes and callus induction media. Biodiversitas 21: 3811-3817. This study aimed to investigate the effects of genotypes and primary callus induction media on somatic embryogenesis of superior robusta coffee clones of Lampung. Leaf explants of clones Tugusari, Komari, Tugino, and Wanto were cultured on two types of primary callus induction media (PCIM). PCIM1 consisted of half-strength MS salts, 30 gL-1 sucrose, added with (mgL-1) 0.1 thiamine-HCl, 0.5 nicotinic acids, 0.5 pyridoxine-HCl, 100 Myo-inositol, 200 ascorbic acids, 150 citric acids, and 1 benzyl adenine. PCIM2 consisted of NPCM salts, 30 gL-1 sucrose, added with (mgL-1) 15 thiamine-HCl, 1 nicotinic acid, 1 pyridoxine-HCl, 2 glycines, 130 Myo-inositol, 200 ascorbic acids, 150 citric acids, 1 2,4-dichlorophenoxyacetic acid, and 2 thidiazuron. The highest percentage (100%) of primary callus formation was found in Komari and Wanto clones. PCIM2 resulted in more primary calli than PCIM1. When subcultured to embryogenic callus induction medium, primary calli of clone Komari and Wanto developed into a high percentage of embryogenic calli, while those of the other two turned brown and died. PCIM2-derived primary calli developed into more embryogenic calli. When subcultured on somatic embryo (SE) regeneration medium, these calli underwent the formation of SE of various stages. When subcultured to plant regeneration medium, these SEs developed into plantlets.

scholarly journals Production high class of micro tuber potato seeds (Solanum tuberosum L.). by Using tissue culture technique

2009 ◽  
Vol 3 (2) ◽  
pp. 99-106
Author(s):  
A.A. Al-jibouri ◽  
A.A. Al-salhay
Keyword(s):  
Tissue Culture ◽  
Solanum Tuberosum L ◽  
Culture Technique ◽  
Potato Cultivars ◽  
Condition Numbers ◽  
Mass Propagation ◽  
High Class ◽  
Dormancy Period ◽  
Elisa Technique

The aim of this investigation was produced micro tubers of four potato cultivars Premiere, Bintje, Estima and Escort in vitro. Apical meristems (0.2-0.4 mm) of potato cultivars were excised and cultured on nutrient medium and incubated at 24±2 Cº and 1000 lux light intensity for 16 hrs per day. The developing plantlets were examined serological by using ELISA technique to eliminate the viral infected plantlets. The virus-free plantlets were chopped into pieces with single bud and re cultured on fresh medium for mass propagation. For micro tubers formation in test tubes, the cultures were transferred to another medium containing a high percent of sucrose (60g/L) with different concentrations of kinetin; the cultures were incubated under 16±2 Cº and 8 hrs photoperiod. The plantlets formed micro tubers after 8-10 weeks from culturing. The results showed significant differences among cultivar’s in their response to in vitro culture and micro tubers formation. The results also showed that the kinetin concentration had significant effect on micro tubers, and 1mg/l kinetin concentration was the best. The micro tubers were stored for 10 week at 4Cº to break down the dormancy period, and gave 100% germination under nursery condition. Numbers of tubers derived from micro tubers and normal tubers of these cultivars were compared at the end of season.

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 Mass Propagation of Feronia limonia L. through Tissue Culture

1970 ◽  
Vol 45 (1) ◽  
pp. 75-78 ◽  
Author(s):  
Shahina Islam ◽  
Mosfequa Zahan ◽  
Shahina Akter ◽  
Tanjina Akhtar Banu ◽  
Ahashan Habib ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Plant Growth ◽  
Key Words ◽  
Multiple Shoot ◽  
Shoot Tips ◽  
Nodal Explants ◽  
Ms Medium ◽  
Ex Vitro ◽  
Mass Propagation

An efficient mass propagation method for Feronia limonia was developed from excised shoot tips and nodal explants of in vitro grown seedlings. Explants were cultured on MS medium with different conc. of NAA, Kn, IAA and BAP singly or in combinations. Highest number of micro shoots and better plant growth were obtained from these two explants on MS medium supplemented with 0.2 mg/l BAP alone. The regenerated shoots were successfully rooted on MS medium supplemented with 0.5 mg/l NAA. The in vitro raised plantlets were successfully established in soil following the formation of roots with 100% survivability under ex vitro condition. Key words: Feronia limonia; Mass propagation; Node; Shoot tips; Multiple shoot DOI: 10.3329/bjsir.v45i1.5186 Bangladesh J. Sci. Ind. Res. 45(1), 75-78, 2010

scholarly journals MICROPROPAGATION OF MUSA ON A STATIONARY LIQUID MEDIUM

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 696g-696
Author(s):  
Joan P. Harris
Keyword(s):  
Liquid Medium ◽  
Mutation Rate ◽  
Multiplication Rate ◽  
In Vitro Growth ◽  
Liquid Media ◽  
Stationary Liquid ◽  
One Year

In vitro growth of banana, Musa sp., using a modified Murashige and Skoog liquid medium was compared with the same formulation solidified by the addition of agar. Plants grown on the stationary liquid weighed more and had a higher multiplication rate. The best volume of liquid and number of plants per vessel were determined. Four hundred plants grown in liquid media were studied for one year after planting in the field. Flowering occurred at 7 months and harvesting 3 months later. The mutation rate observed was below 5%.

scholarly journals Morphological, Histological, and Protein Profiling of Tea Embryo Axis at Early Stage of Culture

2021 ◽  
Vol 6 (3) ◽  
pp. 64403
Author(s):  
Ratna Dewi Eskundari ◽  
Taryono Taryono ◽  
Didik Indradewa ◽  
Yekti Asih Purwestri
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Apical Meristem ◽  
Protein Profile ◽  
Scientific Information ◽  
Protein Profiling ◽  
Root Apical Meristem ◽  
Embryo Axis ◽  
In Vitro Organogenesis

Tissue culture is an alternative choice of plant propagation either through somatic embryogenesis or in vitro organogenesis techniques. TRI2025 tea clone has been cultured successfully, however, the scientific information related to morphology, histology, and protein profile at an early event of culturing time has not been reported yet. This study aimed to determine the differences between those pathways, in the context of morphology, histology, and protein profile. The explants were the embryo axis of TRI2025 tea clone cultured on two different induction mediums; somatic embryogenesis and in vitro organogenesis induction medium. The results showed that most of the explants cultured on A medium developed to be a globular-like structure at 11-day after culture (DAC), while all explants cultured on B medium showed the initiation stage of in vitro organogenesis. Histological analysis showed meristem reconstruction at shoot apical meristem (SAM) and root apical meristem (RAM) at 11-DAC at explants cultured on B medium, while explants cultured on A medium showed callusing at 21-DAC. Protein profile analysis using SDS-PAGE showed protein bands of 54 and 81 KDa that only appeared at explants cultured on A medium start from 14-DAC, and those two protein bands thought to be a differentiator at the early stages of the two tissue culture techniques. Thus, these parameters can be used as early detection for plant tissue culture, especially in tea. 

scholarly journals Coffee Yield and Mineral Cycle in Intercropping of Coffea canephora and Some Species of Timber Shade Trees

2008 ◽  
Vol 24 (1) ◽  
Author(s):  
Adi Prawoto
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Phenolic Compound ◽  
Embryogenic Callus ◽  
Theobroma Cacao ◽  
Coffea Canephora ◽  
Coffee Yield ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Theobroma Cacao L

Cocoa (Theobroma cacao L.) like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D) with kinetin (kin). Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although  the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance

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