scholarly journals PEMBENTUKAN KALUS DAN EMBRIO SOMATIK KAKAO MENGGUNAKAN THIDIAZURON MELALUI SATU TAHAP INDUKSI KALUS

2020 ◽  
Vol 20 (4) ◽  
pp. 179 ◽  
Author(s):  
NUR AJIJAH

<p>ABSTRAK</p><p><br />Embriogenesis somatik kakao (Theobroma cacao L.) telah banyak<br />dilaporkan  dengan  penggunaan  zat  pengatur  tumbuh  (ZPT)  yang<br />bervariasi. Penggunaan thidiazuron untuk menginduksi embriogenesis<br />somatik kakao telah dilaporkan melalui dua tahap induksi kalus. Penelitian<br />ini bertujuan untuk mengevaluasi efektivitas thidiazuron menginduksi<br />embriogenesis somatik kakao melalui satu tahap induksi kalus. Penelitian<br />dilaksanakan di Laboratorium Kultur Jaringan Unit Pengembangan Benih<br />Unggul, Badan Litbang Pertanian, Bogor. Empat taraf thidiazuron (0; 2,5;<br />5,0; dan 10 µg/l) dikombinasikan dengan 2,4-D 2 mg/l<br />digunakan untuk<br />menginduksi kalus dan embrio somatik 3 klon kakao (TSH858, Sca6, dan<br />ICS13) menggunakan eksplan mahkota bunga dan staminoid. Media dasar<br />DKW tanpa ZPT digunakan sebagai kontrol. Penelitian disusun dalam<br />rancangan lingkungan acak lengkap dengan lima ulangan. Setiap unit<br />percobaan terdiri dari sepuluh eksplan. Peubah yang diamati meliputi<br />persentase pembentukan kalus umur 2 dan 4 minggu, penampakan visual<br />kalus, persentase eksplan membentuk embrio somatik, dan jumlah embrio<br />somatik per eksplan umur 10 dan 14 minggu. Kalus terbentuk pada media<br />dengan penambahan hanya 2,4-D atau 2,4-D + thidiazuron, namun embrio<br />somatik hanya terbentuk pada media dengan penambahan 2,4-D +<br />thidiazuron. Pembentukan kalus dan embrio somatik sangat dipengaruhi<br />oleh tipe eksplan dan genotipe. Klon Sca6 lebih responsif dibandingkan<br />TSH858 dan ICS13 dan eksplan staminoid lebih responsif dibandingkan<br />mahkota bunga. Hasil studi ini menunjukkan terdapat pengaruh interaksi<br />yang kuat antara ZPT, genotipe, dan tipe eksplan terhadap pembentukan<br />kalus dan embrio somatik kakao serta tidak terdapat perbedaan hasil yang<br />nyata antara pembentukan embrio somatik melalui satu dan dua tahap<br />induksi kalus.<br />Kata kunci: Theobroma cacao L., genotipe, eksplan, zat pengatur tumbuh</p><p>ABSTRACT</p><p><br />Somatic embryogenesis of cacao (Theobroma cacao L.) has been<br />widely reported with varied of plant growth regulators (PGR) used. The<br />use of thidiazuron in inducing somatic embryogenesis of cacao has been<br />reported through a two-step callus induction. The study aimed to evaluate<br />the effectiveness of thidiazuron in inducing somatic embryogenesis of<br />cacao through a one-step of callus induction. The study was conducted at<br />the tissue culture laboratory of Agricultural Seed Development Unit,<br />Indonesian Agency for Agricultural Research and Development, Bogor.<br />Four levels of thidiazuron (0; 2.5; 5.0; and 10 µg/l) in combination with 2<br />mg/l  2,4-D  were  used  for  inducing  callogenesis  and  somatic<br />embryogenesis of three cacao clones (TSH858, Sca6, and ICS13) using<br />petals and staminoids explants. DKW basal medium without PGR was<br />used as a control. The result showed that callus were formed on medium<br />containing only 2,4-D or 2,4-D + thidiazuron, while embryos were only<br />formed on medium containing 2,4-D + thidiazuron. The formation of<br />callus and somatic embryos were highly affected by explant types and<br />genotypes. Sca6 clone was more responsive than TSH858 and ICS13 and<br />staminoids were more responsive than petals. The results of this study<br />revealed that there was a strong interaction between the PGRs, genotypes,<br />and explant types on the formation of cacao callus and somatic embryos.<br />Results of this study also showed no significant difference between the<br />formation of somatic embryos through one and two steps of callus<br />induction.<br />Keywords: Theobroma cacao L., genotypes, explants, plant growth<br />regulators</p>

2017 ◽  
Vol 11 (1) ◽  
pp. 25-31 ◽  
Author(s):  
Kouassi Kan Modeste ◽  
◽  
Manlé Tokpapon Eliane ◽  
Koné Daouda ◽  
Soumahoro André Brahima ◽  
...  

Author(s):  
Sulistyani Pancaningtyas

Cocoa (Theobroma cacao L.) development using modern breeding techniques can be facilitated by propagation of planting material through somatic embryogenesis. Various factors that may affect embryogenesis are the composition of culture medium and culture condition. Hormone commonly used to initiate the formation of callus is auxin with type 2.4-D (2.4 Dichlorophenoxy acetic acid). The aim of this study was to determine the effect of the addition of 2.4 -D hormoneson the process of cocoa embryogenesis. The treatments were arragged in factorial combination in completely randomized design, which consisted of two factors. Thefirst factor was the concentration of auxin 2,4-D 25 %, 50 %, 75 %, and 100 %; and the second factor was cocoa clones; Sulawesi 01 and Sulawesi 02. The resultshowed that the addition of 2.4-D hormone up to 100% on somatic embryogenesis of cocoa for Sulawesi 01 clone was not significantly different from Sulawesi 02 clone for all parameters. While on the addition of 2.4-D, there was significant difference between Sulawesi 01 and 02. Cocoa embryogenic callus using the addition of 2.4-D (25%-100%) was significantly different from control. Increased concentrations of 2,4-D hormone which is applied onto media would inhibit the formation of the somatic embryo. Addition of 2.4 D 25%, encouraged towards non-embryogenic callus. Keywords: 2.4 Dichlorophenoxy acetic acid, embryogenic callus, somatic embryos, cocoa, medium culture, hormone


2008 ◽  
Vol 43 (10) ◽  
pp. 1433-1436 ◽  
Author(s):  
Thiago Édson Ribeiro da Silva ◽  
Luciana Cardoso Cidade ◽  
Fátima Cerqueira Alvim ◽  
Júlio Cézar de Mattos Cascardo ◽  
Marcio Gilberto Cardoso Costa

The objective of this work was to evaluated a procedure for somatic embryogenesis and regeneration of cacao (Theobroma cacao L.) elite clones. Petal explants from cacao clones TSH 565 and TSH 1188 were cultured on PCG and SCG-2 media, for calli growth. Somatic embryos were formed on the surface of embryogenic calli after transfer to embryo development (ED) medium. Clone TSH 565 showed a higher embryogenic potential than TSH 1188. The best combination of carbon source for embryo induction in ED medium was genotype-specific. Embryogenic callus formations increased in micropore tape-sealed Petri dishes, irrespective of cacao genotype. Mature somatic embryos were successfully converted into plantlets.


2021 ◽  
Author(s):  
Thiago Sanches Ornellas ◽  
Yohan Fritsche ◽  
Edison Cardona Medina ◽  
Miguel Pedro Guerra

Abstract Bamboos are an important worldwide non-timber forest product with current rising interest due to their environmentally friendly applications. Besides the consolidated uses of the sweet shoots and culms for structural uses, Dendrocalamus asper is an imposing ornamental bamboo for horticulture. The present work aimed to establish in vitro calli culture and plant regeneration through somatic embryogenesis starting from young inflorescences of the giant bamboo, D. asper. Pre-anthesis inflorescences were collected, disinfested, and subjected to callus induction on MS basal medium supplemented by 0 µM, 9 µM, 18 µM, 27 µM, and 36 µM of 2,4-D in combination with 9 µM of 2-iP or 9 µM Kin. The different obtained calli types were characterized and subcultured in 0 µM, 4.5 µM, 9 µM, and 18 µM of 2,4-D in combination with 9 µM of both cytokinins for multiplication and differentiation. Additionally, the explant incision and its inoculation orientation onto culture media were tested for callus induction improvement. The 2,4-D was essential for callus induction, and its combination with both cytokinins resulted in embryogenic callus induction and further somatic embryos regeneration. The subsequent reduction of this auxin to 4.5 µM resulted in somatic embryo maturation. Somatic embryos transferred to a plant growth regulator-free medium resulted in plantlet conversion. The present work showed the feasibility of using inflorescences as explants and the efficiency of using the 2-iP in combination with 2,4-D to callus induction and in vitro bamboo plant regeneration through somatic embryogenesis.


2009 ◽  
Vol 12 (17) ◽  
pp. 71-80
Author(s):  
Nam Ngoc Trinh ◽  
Sanh Du Nguyen

On the MS medium containing only 2,4-D, callus induction was inhibited. Moreover, these calli were friable and turned brown after two weeks of culture. the three-week old calli were then transferred to the MS medium containing NAA 0.5 mg/l and kinetin 1 mg/l. The somatic embryos with globular shape appeared after 10 days of culture, while the heart shape, torpedo_shape and cotyledonary_shape embryos appeared successively after 15 days of culture. The abnormal embryos occupied at a rate of 34.3% and rarely germinated to plantlets. On the MS medium without plant growth regulators or only with NAA, somatic embryos could not be induced. On MS medium supplemented with ethephon 3 mg/l somatic embryogenesis from calli was inhibited. Plantlets derived from the eggplant somatic embryos had a survival rate up to 95% when transferred to the pots.


HortScience ◽  
2017 ◽  
Vol 52 (1) ◽  
pp. 142-145 ◽  
Author(s):  
Modeste Kan Kouassi ◽  
Jane Kahia ◽  
Christophe N’guessan Kouame ◽  
Mathias Gnion Tahi ◽  
Edmond Kouablan Koffi

The effect of plant growth regulators on callus and somatic embryogenesis induction in four Cocoa (Theobroma cacao) genotypes was studied. Flower explants were harvested early in the morning and cultured on Driver and Kuniyuki Walnut (DKW) medium supplemented with 1 mg·L−1 of five auxins type (2,4 dichlorophenoxyacetic acid (2,4-D), 3,4 dichlorophenoxyacetic acid (3,4-D), 2,4,5 trichlorophenoxyacetic acid (2,4,5-T), 4-amino-3,5,6-trichloropicolinic acid (picloram), and 3,6-dichloro-2-methoxybenzoic acid (dicamba) in combination with 0.25 or 0.5 mg·L−1 of two cytokinins type (benzylaminopurine (BAP) and 6-furfurylaminopurine [kinetin (Kin)] in a factorial experiment. The plant growth regulators 2,4-D and 2,4,5-T proved to have a broad spectrum action on somatic embryogenesis induction compared with 3,4-D or picloram. There were no significant differences between the two concentrations of cytokinins. However, Kin was found to be more effective in promoting somatic embryogenesis than BAP. Combining 1 mg·L−1 2,4,5-T or 2,4-D with 0.25 mg·L−1 Kin had a broad spectrum action on embryogenesis induction. On the other hand, combining mg·L−1 picloram with 0.5 mg·L−1 Kin or 1 mg·L−1 3,4-D with 0.25 mg·L−1 Kin was only able to induce somatic embryogenesis in a few of the genotypes evaluated. The protocol developed during the current study differs from earleir works as the callus (derived from explants cultured on DKW media) was taken directly to embryo development media as opposed to earlier works in which the callus was taken through a secondary media before being transferred to an embryo development media.


1970 ◽  
Vol 19 (1) ◽  
pp. 89-99
Author(s):  
K. Choudhary ◽  
M. Singh ◽  
M. S. Rathore ◽  
N. S. Shekhawat

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l  of 2,4-D and 0.5 mg/l  of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture  legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)


HortScience ◽  
1990 ◽  
Vol 25 (5) ◽  
pp. 573-576 ◽  
Author(s):  
I.E. Yates ◽  
C.C. Reilly

The influence of stage of fruit development and plant growth regulators on somatic embryogenesis and the relation of cultivar response on somatic embryogenesis and subsequent plant development have been investigated in eight cultivars of pecan [Carya illinoensis (Wangenh.) C. Koch]. Explants from the micropylar region of the ovule were more embryogenic when removed from fruits in the liquid endosperm stage than were intact ovules from less-mature fruits or from cotyledonary segments of more-mature fruits. Explants conditioned on medium containing auxin alone or auxin + cytokinin produced more somatic embryos than medium containing cytokinin alone. Under the conditions of this study, frequency of embryogenesis, as well as the germination of somatic embryos leading to plant development, indicated appreciable variation among cultivars. Plant development was greatest by far from somatic embryos of `Schley' than other cultivars studied.


1970 ◽  
Vol 20 (2) ◽  
pp. 157-170 ◽  
Author(s):  
Richard M.S. Mulwa ◽  
Margaret M.A. Norton ◽  
Robert M. Skirvin

Abundant embryogenic callus was obtained from leaf and floral explants of "Chancellor" grape by continuous culture for 12 weeks on Nitsch and Nitsch basal medium supplemented with 9 μM 2, 4-D + 17 μM IASP + either 1 μM BA or 1 μM TDZ (ECIM) in darkness. They were successfully maintained by a five to six week subculture interval on NN medium containing 2 μM 2, 4-D + 0.2 μM TDZ + 4 μM IASP (LTMM). Near synchronous embryo developed from embryogenic callus on medium containing 10 μM IASP + 8 μM NOA + 1 μM TDZ + 1 μM ABA + 2.5 g/l AC (EDMM).  Individually separated somatic embryos were germinated on both NN and half strength of MS containing 0.5 μM BA + 0.025 μM NAA, respectively; normal plantlet conversion from embryos was low (35%).  Whole fruiting plants were obtained. Aberrant embryo development was characterized by failure to form functional shoot meristems following the initial cotyledon expansion during germination. These observations indicate that the embryo conversion stage of the regeneration is difficult and remains a limiting factor requiring more empirical experimentation for improvement in grape tissue culture.   Key words: Chancellor grape, Regeneration, Somatic embryogenesis   D.O.I. 10.3329/ptcb.v20i2.6895   Plant Tissue Cult. & Biotech. 20(2): 157-170, 2010 (December)


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