Encapsulation of Date Palm Somatic Embryos: Synthetic Seeds

Author(s):  
Shawky A. Bekheet
2014 ◽  
Vol 23 (2) ◽  
pp. 147-155 ◽  
Author(s):  
Dinesh Giri ◽  
Sushma Tamta

This protocol has been developed for somatic embryogenesis in Hedychium spicatum. Simultaneously, a method has also been developed for the production of synthetic seeds by using somatic embryos. Direct somatic embryos were developed on cotyledon explants of zygotic embryos on MS supplemented with high concentration of NAA (20.0 µM). Induction of secondary embryogenesis was best in 2,4-D supplemented medium fortified with activated charcoal. Germination of somatic embryos was enhanced by using GA3. Besides this, round and semi-hard beads of somatic embryos (synthetic seeds) could be produced by using 2% Na-alginate and 100 mM calcium chloride and more than 30% germination of synthetic seeds was achieved in MS. Well acclimated plants produced via somatic embryogenesis and/or synthetic seeds were transferred to field where more than 60% survived. This simple study enabled us to obtain a number of plantlets throughout the year each cycle requiring a short period of time. Besides propagation, this study provided an ex situ method for conservation of this vulnerable Himalayan species.D. O. I.http://dx.doi.org/10.3329/ptcb.v23i2.17506Plant Tissue Cult. & Biotech. 23(2): 147-155, 2013  (December)


2014 ◽  
Vol 24 (1) ◽  
pp. 27-36 ◽  
Author(s):  
Zahida Qamar ◽  
Md. Belal Hossain ◽  
Idrees A. Nasir ◽  
Bushra Tabassum ◽  
Tayyab Husnain

Synthetic seeds of cauliflower cv. Chillout were developed by encapsulating mature somatic embryos in neutral gel media. Somatic embryos were obtained by optimizing callus and cell suspension cultures of cauliflower. Friable, yellowish embryogenic calli were obtained on MS supplemented with 2 mg/l  2,4-D and 0.5 mg/l BAP using hypocotyl as explants, while calli were regenerated in media consisting of 5 mg/l BAP, 2 mg/l Kn and 6 mg/l GA3. Somatic embryo-genesis was induced in cell suspension culture where auxins were removed in successive steps triggering  conversion of globular cells into the heart, torpedo stage (71%) and finally into cotyledonary/somatic embryos (28%). The mature somatic embryos were encapsulated by mixing mature cell suspension with sodium alginate and calcium chloride mixture (1 : 4). Developed synthetic seeds germinated into complete plantlets when placed in neutral gel media.  Germination efficiency of synthetic seeds decreased to about 50 per cent after 12 weeks of storage at 4ºC followed by a rapid decrease to zero per cent after 16 weeks. It was also observed that cauliflower plantlets from synthetic seeds survived successfully when transferred to soil demonstrating  that cauliflower synthetic seeds is a promising step towards their  in vivo direct use. Plant Tissue Cult. & Biotech. 24(1): 27-36, 2014 (June) D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19193


2018 ◽  
Vol 86 (2) ◽  
Author(s):  
Rizka Tamania SAPTARI ◽  
. SUMARYONO

 Date palm (Phoenix dactylifera L.) is the most important crop in the dry areas of the Middle East and North Africa. This palm has been introduced to many countries but has not been grown commercially in Indonesia. Date palm propaga-tion by seeds is easy but its progenies are varied and a half of them are male trees that will not produce fruits. Meanwhile, the propagation by offshoots is impractical and technically difficult. Tissue culture makes it possible to massproduce of genetically identicalsuperior date palms. This research aimed to develop somatic embryogenesis (SE) of date palm using shoot tipand young leaves of date palm seedling as explants. Steps on somatic embryogenesis are explant sterilization, callus initiation and proliferation, somatic embryos induction and maturation, and plantlets matura-tion and rooting. Calli emerged from shoot tip explants after  9 weeks of culture in a modified MS medium supplemented with 10 mg/L 2,4-D, 1 mg/L or  3 mg/L 2-iP, and 1.5 g/L active charcoal. The callus was able to bear somatic embryo in the modified MS medium without hormones. Somatic embryos then developed into plantlets, and roots of plantlets were effectively initiated in the medium supplemented with 0.5 mg/L NAA and 1 mg/L IBA.[Keywords:sterilization,  callogenesis, somatic embryo induction, plantlet rooting, clonal propagation]. Abstrak  Tanaman kurma (Phoenix dactyliferaL.) merupakan tanaman terpenting di wilayah kering Timur Tengah dan Afrika Utara. Palma ini telah menyebar ke banyak negara, namun belum ditanam secara komersial di Indonesia. Perbanyakan kurma dengan biji sangat mudah tetapi turunannya sangat beragam dan setengahnya merupakan tanaman jantan yang tidak berbuah. Perbanyakan dengan anakan (offshoots) secara komersial tidak praktis dan relatif sulit. Kultur jaringan memungkinkan untuk dihasilkan secara massal bibit tanaman kurma varietas unggul yang secara genetik seragam. Penelitian ini bertujuan untuk mengembangkan embriogenesis somatik menggunakan eksplan pucuk tunasdan daun muda dari bibit tanaman kurma. Pengembangan embriogenesis somatik terdiri dari tahap sterilisasi eksplan, inisiasi dan proliferasi kalus, induksi dan maturasi embrio somatik, serta pembesaran dan pembentukan akar planlet. Kalus terbentuk dari eksplan pucuk tunassetelah 9 minggu dikultur pada medium MS modifikasi yang ditambahkan 2,4-D 10 mg/L,  2-iP 1 mg/L atau 3 mg/L, dan arang aktif 1,5 g/L.Kalus berhasil diinduksi menghasilkanembrio somatik pada medium MS modifikasi tanpa penggunaan hormon. Embrio somatik kemudian berkembang hingga menjadi planlet, dan akar planlet secara efektif terinisiasipada medium yang ditambahkan NAA 0,5 mg/L dan IBA1 mg/L.  [Kata kunci :sterilisasi,  kalogenesis, induksi embrio somatik, pengakaran planlet, propagasi klonal].


2016 ◽  
Vol 83 (2) ◽  
Author(s):  
. SUMARYONO ◽  
Rizka T SAPTARI

Abstract         Synthetic seed technology has been developed to combine the practical use of ordinary seeds and clonal plant materials. Single somatic embryos of tea (Camellia sinensis L.) clone GMB 9 were encap-sulated using sodium alginate and CaCl2 at different concentrations to produce synthetic seeds.  Encapsulating matrices with and without somatic embryos were made of WP medium and 20 g/L sucrose with   1, 2 or 3%  sodium alginate and 50 or 100 mM CaCl2. Encapsulating matrices without somatic embryos were then tested its physical characteristics, whereas the capsules with somatic embryos were observed its germination rate and secondary embryo formation every week up to six weeks. The results showed that the concentrations of sodium alginate and CaCl2 were affected significantly the physical characteristics of encap-sulating matrix produced. The sufficient level of hardness and highest germination rate was obtained from 2% sodium alginate and 50 mM CaCl2. Sodium alginate at 1% or less produced soft, leaky and oval encapsulating matrices which were not suitable for synthetic seeds. Sodium alginate 3% and 100 mM CaCl2  produced rounded and very hard encapsulating matrices and inhibited the germination of somatic embryos. Germination rates of tea synthetic seeds and somatic embryos without encapsulation were 5 to 20% after six weeks. Abstrak         Teknologi benih sintetik dikembangkan untuk memadukan kepraktisan penggunaan benih biasa dengan bahan tanam klonal. Embrio somatik tunggal dari tanaman teh (Camellia sinensis L.) klon GMB 9 dienkapsulasi menggunakan natrium alginat   dan CaCl2 untuk membuat benih sintetik. Matriks kapsul dengan dan tanpa embrio somatik dibuat dari medium WP dengan sukrosa 20 g/L dicampur dengan natrium alginat 1, 2, atau 3% dan CaCl2 50  atau 100 mM.  Matriks  kapsul  tanpa embrio somatik kemudian diuji sifat fisiknya, sedangkan matriks kapsul dengan embrio somatik diamati daya kecambah dan pembentukan embrio somatik sekunder setiap minggu sampai dengan enam minggu. Hasil penelitian menunjukkan bahwa konsentrasi natrium alginat dan CaCl2 sangat berpengaruh terhadap sifat fisik matriks kapsul yang dihasilkan. Tingkat kekerasan yang memadai dan menghasilkan persentase perkecambahan yang tinggi diperoleh dengan alginat 2% dan CaCl2 50 mM. Natrium alginat pada konsentrasi 1% atau kurang menghasilkan matriks kapsul yang  lunak, mudah bocor dan berbentuk lonjong sehingga tidak sesuai untuk benih sintetik. Natrium alginat konsentrasi 3% pada larutan CaCl2 100 mM menghasilkan benih sintetik yang bulat, sangat keras dan menghambat perkecambahan embrio somatik teh.  Daya kecambah benih sintetik dan embrio somatik teh tanpa enkapsulasi berkisar  5 - 20% setelah enam minggu. 


2015 ◽  
Vol 3 (1) ◽  
pp. 34-38
Author(s):  
Paritala Vikram ◽  
Ilfah Husna Abdullah Ripain ◽  
Kishore K. Chiruvella ◽  
Mohammed Arifullah

Hempedu Bumi, Andrographis paniculata a popular Malay ethnomedicinal plant used to treat various kinds of diseases like common cold, fever, liver disease and snake bite. Andrographolide a bitter compound present in the plant posses many medicinal properties like anti-inflammatory, antiviral, anti-HIV and promising anticancer activity. Large amount of the compound is being extracted from the plants so there is a need for commercial cultivation of this plant, but seed dormancy proved to be a major obstacle for this process. So production of non-dormant synthetic seeds is an alternative to the dormant natural seeds for the mass cultivation of this plant. Hence the present study is aimed to produce somatic embryos and synthetic seeds using plant tissue culture technique. The zygotic embryos from immature seeds of A. paniculata were inoculated on to MS media containing various auxins (NAA, IBA, 2, 4-D), and cytokinins (BAP, TDZ and Kinetin) for the induction of somatic embryos. Out of all the treatments used, only 1.5 mg/L NAA + 0.75 mg/L Kinetin produced somatic embryos after 6 weeks of culture and the number of somatic embryos per explant was 9.8 ± 0.6. NAA alone produced rhizogenic calli and NAA along with BAP produced hard friable white callus, 2, 4-D alone and 2, 4-D with cytokinins (BAP, Kinetin) produced creamy callus, while IBA alone and in combination with cytokinins (BAP, Kinetin), TDZ resulted in germination of embryos into seedlings. The somatic embryos produced were coated with sodium alginate solution and made into synthetic seeds. The synthetic seeds that have been produced can be used as planting material and for conservation of the plant for future uses.


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