scholarly journals Induksi dan Proliferasi Embriogenesis Somatik In Vitro pada Lima Genotipe Kedelai

2017 ◽  
Vol 44 (3) ◽  
pp. 261
Author(s):  
Adam Saepudin ◽  
Nurul Khumaida ◽  
Didy Sopandie ◽  
Dan Sintho Wahyuning Ardie

ABSTRACT<br /><br />Somatic embryo induction medium was reported to be genotype dependent for soybean. This study was aimed to obtain the optimum medium for embryo somatic induction and proliferation, and to regenerate somatic embryo of five soybean genotypes. Five soybean genotypes (Tanggamus, Anjasmoro, Yellow Biloxi, CG-22-10, and SP-10-4) were used in this study. The research was divided into four steps: (1) embryogenic callus induction of  five soybean genotypes, (2) embryogenic callus proliferation of five soybean genotypes, (3) optimation of embryo somatic induction on five soybean genotypes and (4) embryo somatic regeneration of five soybean genotypes. The induction experiment showed that based on number of embryogenic callus, the best somatic embryo-induction medium was 3% sucrose+ NAA 5 mg L-1+2,4-D 5 mg L-1+ Vitamin B5. Embryogenic callus number for each genotype tested was increased on proliferation media of 3% sukrosa + 2,4-D 5 mg L-1 + NAA 5 mg L-1+ Vit B5, and Yellow Biloxi gave the highest number of proliferated somatic embryos compared to other genotypes. Increasing number of globular somatic embryo of all genotypes was obtained from the optimation of somatic embryo induction media being used, and Tanggamus genotype gave the highest number of globular somatic embryo which followed by Yellow Biloxi genotype. Tanggamus and Yellow Biloxi genotypes were also successfully formed the four steps of somatic embryos (globular, heart, torpedo, and cotyledonary stages), but in regeneration medium of MS0 and media MS + sukrosa 10 g L-1 + GA3 2 mg L-1 + BAP 4 mg L-1 + Vit B5 only Tanggamus genotype was regenerated into plantlet.  <br /><br />Keywords: 2,4-D, NAA, somatic embryos, induction, proliferation<br /><br />

2020 ◽  
Vol 14 (2) ◽  
pp. 73-81
Author(s):  
Yelnititis Yelnititis ◽  

Kulim is one of woody plant that have multifunction as wood source and for spice and medicinal. Generative propagation of this plant have trouble because seed use limited. The use of leaf segment through somatic embryogenesis to solve the problem. The objective of this study is to obtain the best treatment to embryogenic callus induction. The modification of basal medium of Murashige and Skoog was used as growth medium. The experiment was conducted in three stages are callus induction, embryogenic callus and somatic embryo induction. The treatment of 2,4-D (3,0 – 12 mg/l) used for callus induction. For embriogenic callus induction used 2,4-D (3,0 – 12,0 mg/l) combined with NAA 0,5 mg/l. The treatment of thidiazuron (0,1 – 0,7 mg/l) used for somatic embryo induction. The result showed that the treatment of 2,4-D 6,0 mg/l is the best for callus induction with compact of texture, green, dry and non embryogenic. The treatment of combination 2,4-D 12.0 mg/l with NAA 0.5 mg/l is the best for friable callus induction. The treatment of 2,4-D 6.0 mg/l combined with NAA 0,5 mg/l is the best for embryogenic callus induction with very friable of texture, easy to separate, dry, smooth and glossy. Thidiazuron of 0,1 mg/l treatment is the best for somatic embryos induction with the average number of 7,8 somatic embryos.


Author(s):  
Muniappan V ◽  
Manivel P ◽  
Prabakaran V ◽  
Palanivel S ◽  
Parvathi S

Somatic embryogenesis was carried out epicotyl portion of the mature embryo/apical portion. The somatic embryo induction medium containing 2,4-D or NAA (10.0 to 50.0 mg/l). Of the two concentrations tested 2,4-D (30.0mg/l) recorded the highest percentage of response followed by NAA (30.0mg/l). But the highest number of somatic embryo were recorded in 30.0mg/l of 2,4-D followed by NAA. The apical portion of the mature embryo formed direct embryos without any intervention of callus. The maximum percentage of embryogenic cultures were noticed in 30.0mg/l of 2,4-D followed by NAA at 30.0mg/l. for the differentiation of somatic embryos, the embryogenic masses were transferred to medium without any growth regulator. The maximum number of somatic embryos per culture was recorded in 30 mg/l of 2,4-D followed by 30.0 mg/l of NAA. Keywords: Arachis hypogaea L.,Somatic Embryogenesis, 2,4-D and NAA


2017 ◽  
Vol 17 (1) ◽  
pp. 9
Author(s):  
Yosi Zendra Joni ◽  
Riry Prihatini ◽  
Darda Efendi ◽  
Ika Roostika

<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>


2010 ◽  
Vol 140 (2) ◽  
pp. 225-243 ◽  
Author(s):  
Priyanka Saha ◽  
Subhendu Bandyopadhyay ◽  
Sarmistha Sen Raychaudhuri

2018 ◽  
Vol 86 (1) ◽  
Author(s):  
Imron Riyadi ◽  
Darda EFENDI ◽  
Bambang S PURWOKO ◽  
Djoko SANTOSO

AbstractA right combination of cytokinin is able to support the process of callus differentiation to somatic embryo formation in plant somatic embryogenesis. Liquid culture application could increase the efficiency of in vitro culture process on plants. This research aimed to determine the best concentration of TDZ combined with kinetin for callus differentiation to  somatic embryo of sago palm on three culture methods. Plant material used was embryogenic callus derived from tips meristem culture from sucker of Alitir sago palm. Callus was cultured on modified MS media added with: 0.0, 0.1, 0.5 and 1.0 mg/L TDZ combined with 0.5 mg/L kinetin for 12 weeks with subcultures every 6 weeks. Three culture methods used were suspension, temporary immersion system (TIS), and solid media. There were 12 treatments with 4 replicates. The results showed that the highest number of somatic embryos was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin in 6 weeks (167.3 embryos/flask) and 12 weeks (389.2 embryos/flask) with its fresh weight of 18.4 g and 29.1 g, respectively. The highset survival rate in final culture (12 weeks) was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin (100%). The shortest time for somatic embryos expression was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin in two weeks after culture. Histological analysis of early-stage somatic embryos showed the presence of dense and compact cellular arrangements which formed growth spot axis for shoot or SAM (shoot apical meristem) and root or RAM (root apical meristem) that connected each other. [Key words: culture method, embryogenic callus, Metroxylon sagu Rottb., kinetin, sago palm, TDZ]   AbstrakAplikasi kombinasi sitokinin yang tepat dapat mendorong proses diferensiasi kalus membentuk embrio somatik pada proses embriogenesis somatik tanaman. Penggunaan metode kultur cair dapat meningkatkan efisiensi proses kultur in vitro tanaman. Penelitian ini bertujuan untuk menentukan konsentrasi TDZ terbaik dikombinasikan dengan kinetin dalam proses diferensiasi kalus membentuk embrio somatik tanaman sagu pada tiga metode kultur. Bahan tanam penelitian  berupa kalus embriogenik tanaman sagu asal kultur meristem pucuk dari anakan sagu jenis Alitir. Kalus dikulturkan pada media modifikasi dengan penambahan  TDZ dengan konsentrasi 0,1; 0,5; dan 1,0 mg/L dikombinasikan dengan kinetin 0,5 mg/L selama 12 minggu yang disubkultur pada umur 6 minggu. Metode kultur yang digunakan terdiri atas tiga macam yaitu: kultur suspensi, sistem perendaman sesaat (SPS) dan media padat. Perlakuan terdiri atas 12 kombinasi perlakuan dengan empat ulangan. Hasil penelitian menunjukkan bahwa rerata jumlah embrio somatik tertinggi dicapai pada perlakuan metode kultur SPS dengan TDZ 1,0 mg/L baik pada umur kultur 6 minggu (167,3 buah) maupun umur 12 minggu (389,2 buah). Rerata bobot segar tertinggi juga diperoleh pada perlakuan metode kultur SPS dengan TDZ 1,0 mg/L  pada umur kultur 6 minggu (18,4 g) dan  12 minggu (29,1 g). Rerata daya hidup kultur akhir (12 minggu) tertinggi  sebesar 100% diperoleh pada perlakuan SPS. Induksi embrio somatik  tercepat yakni setelah  dua minggu diperoleh pada  metode kultur SPS dengan TDZ 1,0 mg/L dikombinasikan dengan kinetin 0,5 mg/L. Analisis histologi embrio somatik stadium awal  menunjukkan adanya susunan sel yang rapat dan kompak yang menyusun semacam poros atau berkas titik tumbuh tunas atau SAM (shoot apical meristem) maupun akar atau RAM (root apical mersitem) yang saling terhubung.[Kata kunci: kalus embriogenik, metode kultur, kinetin, TDZ, sagu, Metroxylon sagu]


2017 ◽  
Vol 9 (4) ◽  
pp. 352-360
Author(s):  
SUPATMI SUPATMI ◽  
HANI FITRIANI ◽  
NURHAMIDARR RAHMAN ◽  
N. SRI HARTATI ◽  
ENNY SUDARMONOWATI

Supatmi, Fitriani H, Rahman N, Hartati NS, Sudarmonowati E. 2017. Robust in vitro propagation and regeneration of ubi kuning high beta carotene cassava genotype through somatic embryogenesis. Nusantara Bioscience 9: 352-360. Ubi kuning is a local genotype of cassava with high beta carotene content but the development of this genotype is still low because of plant disease susceptibility. Objectives of this study were to robust induce and regenerate somatic embryos of ubi kuning in vitro as well as to define a protocol of cyclic somatic embryogenesis of ubi kuning. Different size of leaf lobes, various concentration of picloram and different light conditions were tested to produce an effective and efficient somatic embryos (SEs).The best response of the induction of embryogenic callus was observed in leaf lobes explant with range size of 1-3 mm and >5mm cultured on induction medium (MS + 4% sucrose + 4 μM CuSO4 + 0.1 mM Glutamine + 0.8% Microagar) supplemented with either 10 or 18 mg/L picloram grown under dark light for 4 weeks. Retransferring embryogenic callus to the same medium supplemented with 16 mg/L picloram gave the advanced development of primary somatic embryos (PSEs) after 70 d grown under both dark and light condition treatments. A positive correlation between globular and cotyledon stages was obtained in all treatments (P≤ 0.01). The highest shoot and root growth (30% and 25%) was achieved in the regeneration of cotyledonary like-tissues cultured on callus embryogenic media (CEM) (MS basal+ 2.5 μM CuSO4 + 3% sucrose + 2.75 g/L phytagel) supplemented with 1.6 mg/L of BAP (6-Benzylaminopurine).


HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1078A-1078
Author(s):  
Qian Zhang ◽  
Jianjun Chen ◽  
Richard J. Henny

Homalomena `Emerald Gem' is an important ornamental foliage plant and widely used for interior plantscaping. Current propagation of this cultivar has been primarily carried out through in vitro culture by organogenesis; regeneration through somatic embryogenesis has not been documented. This report describes successful plant regeneration via direct somatic embryogenesis from explants of different organs. Somatic embryos formed at and around the cut surface of petiole, spathe, and peduncle explants. Embryos also appeared at the base between expanded ovaries of the spadix segment, and around midrib of leaf explants. The optimal treatments for somatic embryo occurrence from petiole, spathe, and peduncle explants were MS medium containing 0.2 mg/L NAA or 0.5 mg/L 2, 4-D with 2.0 mg/L CPPU, and for spadix explants were MS medium with 0.5 mg/L PAA and 2.5 mg/L TDZ. Somatic embryos appeared 6 to 8 weeks after culture and formed large embryo clumps in 3 to 4 months. Somatic embryos produced more secondary embryos and geminated on induction medium. Multiple shoot development and plant regeneration occurred from somatic embryo clusters on MS medium without hormone or with 2 mg/L BA and 0.2 mg/L NAA. The regenerated plants grew vigorously after transplanting to a soilless container substrate in a shaded greenhouse.


Plants ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 38 ◽  
Author(s):  
Ghulam Raza ◽  
Mohan B. Singh ◽  
Prem L. Bhalla

The efficient regeneration of plants from commercial genotypes is a pre-requisite for successful genetic transformation, to apply modern crop improvement techniques such as CRISPR-based genome editing. Plant regeneration through the somatic embryogenesis pathway offers an advantage over the organogenesis approach, avoiding the risk of developing chimeras. Plant genotype, explant type, and media compositions play an essential role in the in-vitro regeneration of plants. This study aimed to characterize the commercially grown Australian soybean genotypes for their potential to induce somatic embryos, embryo proliferation, maturation, germination, and plant regeneration. Overall, nine soybean cultivars belonging to different maturity groups were evaluated. Immature cotyledon ranging from 2–4 and 4–6 mm in size were used as explants for somatic embryogenesis induction. Maximum somatic embryo induction frequency (86%) was observed from 4–6 mm immature cotyledons of the cv. Jack (MG III), followed by 66%, 26%, 21%, and 6% in cultivars Williams (MG III), Snowy (MG III), MoonB1 (MG V), and PNR791 (MG V), respectively. On the other hand, cv. Snowy showed maximum somatic-embryo-inducing potential (67%) in 2–4 mm immature cotyledons followed by Williams, Jack, MoonB1, and PNR791. Somatic embryos from Jack, Williams, and Snowy cultivars were further tested for embryo proliferation, maturation, and germination. Maximum proliferation and maturation were observed in cv. Jack, followed by Snowy and Williams. However, cv. Snowy showed a significantly higher conversion of cotyledonary stage embryos to plantlets (85%), than both Jack and Williams cultivars (53% each). In conclusion, this study outlined a protocol for somatic embryogenesis and plant regeneration from three soybean cultivars. Our findings suggest commercial cv. Snowy could be a good candidate for developing transgenic plants through somatic embryogenesis.


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