scholarly journals Plant regeneration from hypocotyls of black carrot via direct somatic embryogenesis and determination of its genetic stability by RAPD and iPBS methods

Author(s):  
Burcu Çetin

A study was carried out to investigate plant regeneration from hypocotyls of black carrot (Daucus carota spp. sativus), an industrially and medicinally important plant via somatic embryogenesis, and the genetic stability of obtained plantlets with the random amplified polymorphic DNA (RAPD) and inter primer binding sites (iPBS) methods. Hypocotyl explants isolated from in vitro germinated seeds were incubated initially at Murashige and Skoog (MS) medium containing 0.5 mg/L or 1 mg/L 2,4-D for 2 weeks and then at hormone-free MS medium in order to obtain somatic embryos, which were observed after 15 days on the explants. Performed molecular analyses showed that all bands obtained from plantlets through micropropagation were monomorphic. This protocol where black carrot was determined to be clonally reproduced through somatic embryogenesis may lead the way for further research regarding germplasm conservation and breeding studies.

Author(s):  
Ghan Singh Maloth ◽  
Rajinikanth Marka ◽  
Rama Swamy Nanna

In the present study it was reported on direct somatic embryogenesis and plant regeneration from cotyledon and leaf explants of Turkey berry/pea egg plant (Solanum torvum SW), a medicinally important plant. Somatic embryogenesis has several advantages over other routes of in vitro plant regeneration. Somatic embryogenesis was induced directly from cotyledon and leaf explants on MS medium fortified with BAP (0.5 mg/L)+NAA (0.5-6.0 mg/L). High percentage of somatic embryogenesis (90%), maximum number of somatic embryos formation (62±0.18)  along with high percentage (76%) conversion of somatic embryos into bipolar embryos was observed on cotyledon explants in 0.5 mg/L BAP+2.5 mg/L NAA. At the same concentration of BAP (0.5 mg/L)+NAA (2.5 mg/L) also resulted  on the maximum percentage of somatic embryogenesis (92%), the highest number of somatic embryos formation (88±0.15) and the highest percentage (76%) of somatic embryos conversion into bipolar embryos in leaf explants. A mixture of globular, heart and torpedo-shaped embryos were germinated on MS medium supplemented with 0.5 mg/L IAA+1.0-4.0 mg/L BAP. Maximum germination frequency (75±0.14) of somatic embryos and plantlet formation was found in 0.5 mg/L IAA+2.0 mg/L BAP, but they didn’t germinate on ½ MSO and MSO media. The survival rate of regenerated plants after field transfer was recorded to be 75%. These regenerated plants were found morphologically similar to donor plants. The present protocol can be used for conservation of the species and also for genetic transformation experiments in S. torvum.


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.


2016 ◽  
Vol 24 (1) ◽  
pp. 29-36 ◽  
Author(s):  
Jelili Opabode ◽  
Oluyemisi Akinyemiju

Abstract An efficient and reproducible micropropagation protocol of Solanecio biafrae (Oliv. & Hiern) C. Jeffrey has been developed from nodal stem segments. Shoot development was obtained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP) alone and in combination with zeatin and 1-naphthaleneacetic acid (NAA). Elongated shoots were rooted in the presence of zeatin or 3-indole-butyric acid (IBA) alone or in combinations. The highest number of explants forming shoots (100%) as well as the highest number of shoots per explant (3.4) and the longest shoots (22 mm) were recorded on medium containing 4.0 mg·dm−3 BAP, 2.0 mg·dm−3 NAA, and 1.0 mg·dm−3 zeatin. About 76% of shoots formed roots on half-strength MS medium free of plant growth regulators. The best root formation (approximately 88%) was recorded on the medium containing 1.0-1.5 mg·dm−3 IBA. The micropropagated shoots with well-developed roots were efficiently acclimatized under greenhouse conditions. The random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) amplification products were monomorphic in micropropagated plants and similar to those of mother plant showing their genetic uniformity. This is the first report of micropropagation of S. biafrae, which will facilitate in vitro mass propagation, conservation, and germplasm exchange of this endangered African vegetable.


2017 ◽  
Vol 17 (2) ◽  
pp. 91
Author(s):  
Meutia Zahara

Abstract Phalaenopsis orchids are recognized as the most popular orchid genus in the world, especially in horticultural industry due to their large, colorful, and durable flowers as well as their wider adaptability to room conditions. The characteristics of seedling propagated by vegetative means are not uniform; therefore, propagation through tissue culture is desirable. Although the micro propagation of Phalaenopsis has shown very good development, but the wide spread of micro propagation still limited due some problems such as the exudation of phenolic compounds, the PGR concentration, the media used, somaclonal variation, the chosen explants, etc. This paper endeavor to include some important investigations based on the common explants used; leaf and flower stalk. Keywords: Micropropagation, Phalaenopsis, leaf explant, flower stalk ReferencesAnonymous. Orchid (Orchidaceae). Diakes tanggal 13 Januari 2013 dari http://www.rainforest-alliance.org/kids/species-profiles/orchid. Rainforest Alliance. 2002.Pillon, Y.; Chase, M. W.Taxonomic exaggeration and its effects on orchid conservation. Conservation Biology. 2007, 21, 263–265.Thengane, S. R.; Deodhar, S. R.; Bhosle, S. V.; Rawal, S. K. Direct somatic embryogenesis and plant regenaration in Garciniaindica Chois’. Current Science. 2006, 91(8), 1074-1078.Yuswanti, H.; Dharma, I. P.; Utama. ; Wiraatmaja, I. W. Mikropropagasi anggrek Phalaenopsis dengan menggunakan eksplan tangkai bunga. AGROTROP. 2015, 5(2): 161-166.Raynalta, E.; Sukma, D.  Pengaruh komposisi media dalam perbanyakan protocorm like bodies, pertumbuhan plantlet, dan aklimatisasi Phalaenopsis amabilis. J. Hort. Indonesia. 2013, 4(3): 131-139.Kosir, P.; Skof, S.; Luthar, Z. Direct Shoot Regeneration from Nodes of Phalaenopsis of Orchids. Acta Agriculturae Slovenica. 2004, 83, 233–242.Arditti, J. R. ; Ernst. Micropropagation of Orchids. Wiley-Interscience. New York, 1993.Park, Y. S.;Kakuta, S.; Kano, A.; Okabe, M.Efficient propagation of protocorm-like bodies of Phalaenopsis in liquid medium. Plant Cell, Tissue and Organ Culture. 1996, 45, 79–85.Park, S. Y. ; Yeung, E. C.; Chakrabarty, D. ; Paek, K. Y. An efficient direct induction of protocorm-like bodies from leaf subepidermal cells of Doritaenopsis hybrid using thin-section culture. Plant Cell Reports. 2002, 21, 46–51.Zahara, M.; Datta, A.; Boonkorkaew, P. Effects of sucrose, carrot juice and culture media on growth and net CO2 exchange rate in Phalaenopsis hybrid ‘Pink’. ScientiaHorticulturae. 2016,205, 17–24.Hee, K. H.; Loh, C. S.; Yeoh, H. H. In vitro flowering and rapid in vitro embryo production in Dendrobium Chao Praya Smile (Orchidaceae). Plant Cell Reports. 2007, 26, 2055–2062.Kannan, N. An in vitro study on micropropagation of Cymbidium orchids. Current Biotica. 2009, 3, 244–250.Steward, Jr. N. C. Plant Biotechnology and Genetics. Willey, A john Willey & Sons, INC., Publication. 2008.George, E. F.; Sherington, P. D.Biotechnology by tissue culture. Exegetics Ltd. 1994.Nursyamsi. Teknik kultur jaringan sebagai alternatif perbanyakan tanaman untuk mendukung rehabilitasi lahan. Makalah pada ekspose hasil-hasil penelitian balai penelitian kehutanan makasar. Makasar, 2010.Aditi, J. F. L. S.; Krikorian, A. D. Orchid mircropropagation: the path from laboratory to commercialization and an account of several unappreciated investigators. Botanical Journal of of the Linnean Society. 1996, 122: 183-241.Gunawan, L. W. Teknik Kultur Jaringan Tanaman. Pusat Antar Universitas (PAU) Bioteknologi IPB. 1998. Bogor.Chugh, S. Guha, S.; Rao, I. U. Micropropagation of orchids: A review on the potential of different explants. Scientia Horticulturae. 2009, 122, 507–520.Ramdan. Kultur daun dan pangkal batang in vitro anggrek bulan raksasa (Phalaenopsis gigantea J.J.Smith) pada beberapa media kultur jaringan. Departemen agronomi dan hortikultura, Fakultas pertanian IPB. 2011.Latip, M. A. R.; Murdad, Z. A.; Aziz, L. H.; Ting, L. M.; Govindasamy.; R. Pipin. Effects of N6-Benzyladenine and Thidiazuron on Poliferation of Phalaenopsis gigantea Protocorm. AsPac J. Mol. Biol. Biotechnol. 2010, 18(1): 217-220 p.Niknejad, A.; Kadir, M. A.; Kadzimin, B. S. In vitro plant regeneration from protocorms-like bodies (PLBs) and callus of Phalaenopsis gigantea (Epidendroidaceae: Orchidaceae). African Journal of Biotechnology.2010, 10, 11808–11816.Chen, J. T.; Chang, W. C. Direct somatic embryogenesis and plant regeneration from leaf explants of Phalaenopsis amabilis. Biologia Plantarum. 2006, 50, 169–173.Zahara, M. Disertasi doktor: The Effects of Plant Growth Regulators and Natural Additives on Direct Shoot Regeneration and Plantlet Growth of Phalaenopsis hybrid ‘Pink’. Asian Institute of Technology, Pathumthani. Thailand. 2016.Xu, C. J.; Li, H.; Zhang, M. G. Preliminary studies on the elements of browning and the changes in cellular texture of leaf explant browning in Phalaenopsis. Acta Horticulturae Sinica. 2005, 32, 1111–1113.Tokuhara, K; Mii, M. Induction of embryonic callus and cell suspension culture from shoot tips excised from flower stalk buds of Phalaenopsis (Orchidaceae). In Vitro Cellular & Developmental Biology–Plant. 2001, 37, 457–461Balilashaki, K.; Naderi, R.; Kalantari, S.; Soorni, A. Mircropropagation of Phalaenopsis amabilis cv Cool ‘Breeze’ with using flower stakl nodes and leaves of sterile obtained from node cultures. IJFAS, 2014.Semiarti, E.; Indrianto, A.; Purwanto, A. Agrobacterium-Mediated transformation of Indonesian orchids for  micropropagation, genetic transformation, Prof. MarÃa Alvarez (Ed.), ISBN: 978-953-307-364-4, InTech, 2011. Available from: http://www.intechopen.com/books/ genetic-transformation/agrobacterium-mediated-transformation-ofindonesian-orchids-for-micropropagation.


2008 ◽  
pp. 22-33 ◽  
Author(s):  
Marilyn Belarmino ◽  
Jocelyn Gonzales

A study was conducted to establish a reliable procedure for somatic embryogenesis and plant regeneration from callus cultures of purple food yam (Dioscorea alata L.). The procedure involved three steps; (1) culture of nodal stem segments from greenhousegrown plants to generate in vitro plantlets; (2) induction of callus from the leaf, petiole and nodal stem tissues; and (3) initiation of somatic embryo from callus. Results showed that the agar-solidified Murashige and Skoog (MS) medium containing 30 gl-1 sugar, 0.1 gl-1 α-cysteine , 10 mgl-1 calcium pantothenic acid, 2.0 mgl-1 asparagine, 2.0 mgl-1 arginine, 80.0 mgl-1 adenine sulfate (AdSO4) and 0.1 mgl-1 naphthalene acetic acid (NAA) effectively broke dormancy of lateral buds of nodal stem cultures from both ‘VU-2’ and ‘Kinampay‘ varieties. Production of multiple adventitious shoots occurred after transfer of in vitro nodal pieces to the same medium added with 1.0 mgl-1 benzylamino purine (BAP) or, MSA medium. Callus was effectively induced from the vegetative tissues in MS medium added with 1.0 mgl-1 2,4-Dichlorophenoxy acetic acid (2,4-D) or, with picloram. Among the three types of explants, the nodal stem was the most suitable which produced purplish nodular embryogenic callus. A higher percentage of nodal stem-derived calli produced globular embryos in MS medium containing 1.0 mgl-1 2,4-D and 0.5 mgl-1 BAP, or in 1.0 mgl-1 picloram and 0.5 mgl-1 BAP than, in the plant growth regulator-free medium (control). The maturation of embryos was facilitated by one-month culture in MS medium containing 0.1 mgl-1 ABA and 100 mgl-1 glutamine. This step improved the germination of somatic embryos in one-half strength PGR-free MS medium containing 100 mgl-1 glutamine (regeneration medium). All somatic embryoderived plantlets were morphologically normal and established well in soil.


Author(s):  
D. Bele ◽  
Nishi Mishra ◽  
Sushma Tiwari ◽  
M. K. Tripathi ◽  
G. Tiwari

Nodal segments of sandalwood were cultured on MS medium amended with different plant growth regulators in varying concentrations to search out higher in vitro response leading to plantlet regeneration via somatic embryogenesis and/or organogenesis. Higher proportion of direct somatic embryogenesis, number(s) of somatic embryo per explant and plantlet regeneration via direct organogenesis were recorded on MS medium supplemented with a moderate concentration of TDZ (1.0 mgl-1) in combination with comparatively a lower concentration of NAA (0.5 mgl-1). A relative higher concentration of BAP (1.0-2.0 mgl-1) in combination with a lower concentration of NAA (0.5 mgl-1) promoted frequency of indirect somatic embryogenesis. Ratio of organ formation directly from surface of cultured explants was recovered from culture medium fortified with a higher concentration of BA at the concentration of 4.0 mgl-1 in combination with a lower concentration of NAA (0.5 mgl-1). Maximum plantlets regenerated via somatic embryogenesis (direct and/or indirect) on regeneration medium supplemented with 2.0 mgl-1TDZ  in combination with 1.0 mg l-1GA3, while plantlets in higher frequencies via indirect organogenesis was attained with regeneration medium amended with comparatively lower concentration of TDZ (1.0 mg l-1) in combination with 0.5 mgl-1 GA3 and 0.5     mgl-1 NAA. The plantlets were transferred to pots and hardened in Environmental Growth Cabinet and Net House during preliminary weaning period and transferred to field successfully. Morphologically normal plants were recovered.


HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 694c-694
Author(s):  
Sung-Do Oh ◽  
Won-Seob Song ◽  
Man-Sang Lee

From one week through 7 weeks after artificial pollination, immature ovules of yooza(Citrus junos Sieb. et Tanaka) were excised and cultured in vitro on MT media. Even though there was only a little difference in percentage of somatic embryo formation depending upon the time of excision, immature ovules of 4-week-old showed the highest ratio of somatic embryo formation without callus outgrowth. Various growth regulators or other stimulators were added to the MT media to increase the somatic embryogenesis, In general, BAP was more effective than 2,4-D for somatic embryo formation and the combinations of 0.01mg/l 2,4-D and 0,01 or 0.1mg/l BAP were particularly effective in stimulating somatic embryo formation. When 500mg/l malt extract was added to the medium, the percentage of somatic embryo formation increased reaching as high as 86.7%. Plant regeneration from somatic embryos reached to 66.7% on the medium containing 1.0mg/l zeatin. Isozyme banding patterns were also analyzed to confirm the variations of characteristics of the plantlets derived from direct somatic embryos.


2017 ◽  
Vol 29 (2) ◽  
pp. 143-153
Author(s):  
Anna Pindel

AbstractThe present study has described a simple protocol for efficient plant regeneration of Asparagus densiflorus ‘Sprengeri’ and ‘Myriocladus’ using single-node spear explants, and indirect organogenesis via callogenesis induced on internode explants. The results showed that the genotypes ‘Sprengeri’ and ‘Myriocladus’ regenerated to complete plants via nodal cultures and callus tissue, but the plant regeneration response was higher in secondary explants on MS medium with NAA + kinetin (1+1 mg dm-3) after transfer onto a multiplication medium with IAA+BAP (1+4 mg dm-3), and then onto a rooting medium supplemented with IBA (10 mg dm-3) or NAA + kinetin (1+1 mg dm-3). Primary explants of both cultivars showed high regenerative potential (via the callus stage) on MS medium with IAA+BAP. The cultivar Sprengeri also regenerated via somatic embryogenesis. Both kinds of ‘Meyeri’ explants have a morphogenetic potential for the formation of shoots, which, however, were not capable of rooting. This confirms that the explant, genotype and culture medium are determining factors in the in vitro plant regeneration system.


Sign in / Sign up

Export Citation Format

Share Document