scholarly journals Coconut Tissue Culture: The Indian Initiatives, Experiences and Achievements

CORD ◽  
2017 ◽  
Vol 33 (2) ◽  
pp. 11
Author(s):  
Anitha Karun
Keyword(s):  
Tissue Culture ◽  
Somatic Embryos ◽  
Culture Media ◽  
Zygotic Embryo ◽  
Shoot Meristem ◽  
Direct Organogenesis ◽  
Zygotic Embryos ◽  
Immature Inflorescence ◽  
High Yielding Varieties

Coconut is one of the principal crops of India cultivated in over 35 districts mainly in the southern states. The productivity of the crop is declining in many of the traditionally cultivated regions owing to ageing plantations as well as biotic and abiotic stresses. These plantations are to be replanted with high yielding varieties/hybrids for which adequate quantity of quality planting material is not available. Even though tissue culture research was initiated in many laboratories in the country, the work was eventually phased out in most of the laboratories for want of a repeatable protocol.  At ICAR-CPCRI, coconut tissue culture programs have been continuing for the past three decades. The attempts made include experimentation with different explants viz., immature inflorescence, plumular tissues, mature palm shoot meristem, ovary and anthers and different culture media supplemented with varying levels and types of hormones. Some of the successful protocols developed at the Institute include coconut zygotic embryo culture for collection and exchange of germplasm, cryopreservation and retrieval of zygotic embryos and pollen and plantlet regeneration from plumular tissues. Even though ICAR-CPCRI has succeeded in obtaining plantlets via direct organogenesis from inflorescence explants, the absence of friable calli formation from explants, the low rate of somatic embryo formation, large number of cultures turning to abnormal shoot development, non conversion of somatic embryos into plantlets, and formation of abnormal somatic embryos remain the major bottlenecks. Gene expression studies are being currently undertaken to decipher the molecular basis of in vitro recalcitrance.

scholarly journals Can Agricultural Biotechnology Help Guava Growing in Temperate Climate

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 861B-861
Author(s):  
Bipul K. Biswas* ◽  
Nirmal Joshee ◽  
Anand K. Yadav
Keyword(s):  
Tissue Culture ◽  
Somatic Embryos ◽  
Adventitious Shoots ◽  
Multiple Shoots ◽  
Cold Climate ◽  
Nodal Explants ◽  
Temperate Climate ◽  
Zygotic Embryos ◽  
Ms Medium

Guava (Psidium guajava L.), also called `apple of tropics,' is immensely nutraceutical and horticulturally important. Being a tropical plant, it cannot stand temperatures below 25° F and needs frost protection to grow in temperate regions. To adapt in cold climate, cold hardy guava cultivars are needed. Conventional ways are uneconomic in time and efforts. Still, transgenic plants developed using biotechnological approaches of tissue culture and rDNA technology, appear to have great potential. Thus, protocols for in vitro propagation of guava were developed via organogenesis and somatic embryogenesis using nodal explants from mature trees and young zygotic embryos, respectively. Nodal explants induced multiple shoots when cultured on MS medium fortified with KIN, BAP and Ad.S. Adding a (NO3)2 to medium was useful to prevent in vitro shoot tip browning of adventitious shoots. Rocker liquid culture greatly increased growth of multiple shoots compared to the agar-based medium. It appears to be a good tool for woody plant tissue culture. Induction of somatic embryos in guava was also achieved on MS medium supplemented with IAA auxin. About 80% to 90% somatic embryos germinated normally. To achieve Agro-bacterium-mediated gene transfer in guava, on-going co-cultivation of organogenic tissues of guava is to optimize protocols for freeze tolerance gene (CBF1, CBF2, CBF3) transfer. Plasmid vectors containing selectable markers (nptII gene for antibiotic selection and GUS reporter gene as scorable gene mediated selection), with CaMV 35S promoter gene has been introduced into guava tissues and the resultant plants showed antibiotic resistance. Details of the experimental procedures and up-to-date results will be discussed.

scholarly journals In Vitro Flowering of Plantlets Regenerated from Zygotic Embryo-derived Somatic Embryos of Ginseng

HortScience ◽  
1990 ◽  
Vol 25 (12) ◽  
pp. 1652-1654 ◽  
Author(s):  
Haeng S. Lee ◽  
Jang R. Liu ◽  
Seung G. Yang ◽  
Young H. Lee ◽  
Kwang-W. Lee
Keyword(s):  
Somatic Embryos ◽  
Zygotic Embryo ◽  
Ginseng Root ◽  
Zygotic Embryos ◽  
Ms Medium ◽  
Root Explants ◽  
Half Strength ◽  
Dichlorophenoxy Acetic Acid ◽  
Somatic Embryo Induction

Mature zygotic embryos dissected from ginseng (Panax ginseng C.A. Meyer) seeds were cultured on Murashige and Skoog (MS) medium containing various concentrations of 2,4-D and kinetin. Somatic embryos were induced directly from cotyledonary tissue and from intervening callus. The frequency of somatic embryo induction was up to 55% of zygotic embryo explants. Upon transfer onto half-strength MS medium supplemented with 1 mg BA/liter and 1 mg GA3/liter, most somatic embryos developed into plantlets. More than 50% of the plantlets flowered after 4 weeks of culture, and some developed immature fruits in vitro. These results indicate that adulthood of ginseng root explants is not a prerequisite for flowering of plantlets regenerated through somatic embryogenesis. Chemical names used: (2,4 -dichlorophenoxy) acetic acid (2,4-D); N-(2-furanylmethyl) -1H-purin-6-amine(kinetin); N-(phenylmethyl) -1H-purin-6-amine (BA); gibberellic acid (GA3).

DEVELOPMENT OF NEOLAMARCKIA CADAMBA (KELEMPAYAN) TISSUE CULTURE TECHNIQUES FOR SUSTAINABLE SUPPLY OF PLANTING MATERIALS FOR COMMERCIAL PLANTATION

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Siti Suhaila A. Rahman ◽  
Norwati Muhammad ◽  
Nor Hasnida Hassan ◽  
Haliza Ismail ◽  
Nazirah Abdullah ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Mass Production ◽  
Nodal Segments ◽  
Direct Organogenesis ◽  
Timber Species ◽  
Culture Techniques ◽  
Commercial Plantation ◽  
Planting Materials ◽  
Tissue Culture Techniques

Neolamarckia cadamba (kelempayan) is a multipurpose and fast growing timber species. The tree is grown for timber, paper-making and as ornamental plant. It is reported that its barks and leaves possesed medicinal values and its flowers are used in perfumes. The species is also known to be suitable for plywood, packing case, toys and short-fibred pulp. Therefore, mass production of high quality planting material of N. cadamba is important to support plantation program of this species. Here we presented mass production of N. cadamba through tissue culture techniques. Nodal segments derived from in vitro germinated seeds were used and induced direct organogenesis to produce shoots and roots using MS media (1962) and plant growth regulators (BAP and IBA) that are relatively cheaper than previously used methods. The tissue culture technique of N. cadamba developed may help in ensuring supply of planting materials that are feasible for commercial plantation purposes.

scholarly journals Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

2008 ◽  
Vol 43 (10) ◽  
pp. 1325-1330 ◽  
Author(s):  
Lucymeire Souza Morais-Lino ◽  
Janay Almeida dos Santos-Serejo ◽  
Sebastião de Oliveira e Silva ◽  
José Raniere Ferreira de Santana ◽  
Adilson Kenji Kobayashi
Keyword(s):  
Plant Regeneration ◽  
Suspension Culture ◽  
Cell Suspension ◽  
Cell Suspension Culture ◽  
Somatic Embryos ◽  
Culture Media ◽  
Ms Medium ◽  
Regenerated Plants ◽  
Male Flowers

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.

scholarly journals A Technique for the Cultivation of Insect Tissues

1928 ◽  
Vol 6 (1) ◽  
pp. 1-11
Author(s):  
J. G. H. FREW
Keyword(s):  
Tissue Culture ◽  
Culture Media ◽  
Body Fluids ◽  
The Body ◽  
Blow Fly ◽  
In Vitro Tissue Culture ◽  
Successful Technique

In vitro tissue culture Is shown to be a possible mode of experimentation with the tissues of the Blow Fly larva. Methods are described- whereby the tissues, and the body fluids requisite as culture media may be obtained free from bacteria. The imperfections of the technique are noted and the conclusion reached that a successful technique must depend on the rearing of bacteria-free larvae, for which a method Is briefly outlined. It Is shown that progress in this part of the work must await further physiological knowledge, particularly in respect to the nature of the body fluids.

scholarly journals High-frequency Direct Somatic Embryogenesis and Plantlet Regeneration from Leaves Derived from In Vitro-Germinated Seedlings of a Coffea arabica Hybrid Cultivar

HortScience ◽  
2016 ◽  
Vol 51 (9) ◽  
pp. 1148-1152 ◽  
Author(s):  
Jane Kahia ◽  
Margaret Kirika ◽  
Hudson Lubabali ◽  
Sinclair Mantell
Keyword(s):  
Tissue Culture ◽  
High Frequency ◽  
Somatic Embryos ◽  
Coffea Arabica ◽  
Alternative Methods ◽  
Direct Somatic Embryogenesis ◽  
Ms Medium ◽  
Embryogenic Cultures ◽  
Half Strength

Breeding work carried out during the period 1971–85 by the Coffee Research Institute, Ruiru, Kenya resulted in the release of a new improved hybrid Coffea arabica named Ruiru 11. The cultivar combines resistance to coffee berry disease (CBD) and leaf rust, with high yield and good cup quality attributes. The propagation by F1 hybrid seeds production, cuttings, and tip grafting do not produce enough planting materials. There was a need to explore alternative methods and tissue culture offers potential options. The objective of the study was to evaluate the effect of explant sources and cytokinins on induction and regeneration of somatic embryos. Eight different explants were cultured on half-strength Murashige and Skoog (MS) medium supplemented with 10 µm benzylaminopurine (BAP). The effect of kinetin, N6-(2-isopentyl) adenine (2iP) evaluated at (0, 0.5, 5, or 25 µm) or thidiazuron (TDZ) (0, 0.5, 1.0, or 5 µm) added in separate experiments was also evaluated. The percentage of embryogenic cultures and the numbers of embryos per explant were determined after 3 months’ culture. The explant type had a significant effect (P > 0.05) on the induction of somatic embryos. Explants from in vitro-germinated seedlings produced the highest embryogenic cultures (90%) and the highest mean number of embryos (19.36) per explant. Cytokinins strongly enhanced induction and regeneration of somatic embryos. TDZ at 1 µm produced the highest embryogenic cultures (100%) and the highest mean number of embryos (24.2). The embryos were germinated on half-strength MS medium without any hormones. A high (98%) survival rate of the regenerated plantlets was recorded over all the treatments in the greenhouse. This is the first report on induction of high-frequency direct somatic embryos from coffee juvenile tissues. This is of great significance in tissue culture and indeed molecular biology manipulations because it allows regeneration of coffee from several explants.

scholarly journals Cycads in vitro

2015 ◽  
Vol 24 (2) ◽  
pp. 287-301 ◽  
Author(s):  
Jaime A Teixeira Da Silva ◽  
Wynston Ray Woodenberg ◽  
Songjun Zeng
Keyword(s):  
Mass Production ◽  
Germplasm Conservation ◽  
Leaf Tissue ◽  
Habitat Destruction ◽  
Direct Organogenesis ◽  
Zygotic Embryos ◽  
In Vitro Production ◽  
Mature Trees ◽  
Initial Culture

The Cycadales are a group of botanical and evolutionary importance; however, many species face the threat of extinction due to poaching and habitat destruction. The current investigation reviews previous work on in vitro production of cycads, which holds great potential for mass production and germplasm conservation of these unique plants. Megagametophytes and zygotic embryos have been used as explants in most studies, while seedling tissue and new leaf tissue of mature trees have also been used. Callus, coralloid roots and somatic embryos have been formed in vitro but direct organogenesis appears to be the most promising method for mass production and germplasm preservation of the cycads, as recent studies have reported the acclimatization of numerous plantlets less than 200 days after initial culture of zygotic embryos.Plant Tissue Cult. & Biotech. 24(2): 287-301, 2014 (December)

Two alternative pathways of somatic embryo origin from polyembryonic mature stored seeds of Pinus koraiensis Sieb et Zucc.

1997 ◽  
Vol 75 (3) ◽  
pp. 509-512 ◽  
Author(s):  
P. V. Bozhkov ◽  
I. S. Ahn ◽  
Y. G. Park
Keyword(s):  
Somatic Embryo ◽  
Somatic Embryos ◽  
Zygotic Embryo ◽  
Pinus Koraiensis ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Strong Negative Correlation ◽  
Alternative Pathways ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryo Origin

Individual mature stored seeds of Pinus koraiensis sometimes contain several viable zygotic embryos originated through the processes of simple and cleavage polyembryony. To induce the embryonic process, isolated zygotic embryos were cultured on five different media all supplemented with 10 μM 2,4-dichlorophenoxyacetic acid and 5 μM 6-benzyladenine. Two alternative pathways of somatic embryo origin were revealed. The first pathway was associated with the production of a friable, translucent callus in the hypocotyls–cotyledon region of the dominant zygotic embryo. The second pathway was related to the proliferation of a translucent, moist, and mucilaginous tissue (termed embryonal–suspensor mass) in the suspensor region of the dominant zygotic embryo. Both types of tissues contained early somatic embryos. Regression analysis has shown a strong negative correlation between the frequencies of formation of embryogenic callus and embryonal–suspensor mass both at 3 and 8 weeks of culture (r = − 0.85; p = 0.07 and r = −0.71; p = 0.17, respectively). Key words: Pinus koraiensis; polyembryonal seeds; somatic embryogenesis; embryogénie callus; embryonal–suspensor mass.

Histological studies of cellular differentiation during somatic embryogenesis of coconut plumule-derived calli

2015 ◽  
Vol 43 (3) ◽  
Author(s):  
K. Lakshmi Jayaraj ◽  
U. Bhavyashree ◽  
T.P. Fayas ◽  
K.K. Sajini ◽  
M.K. Rajesh ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Callus Formation ◽  
Histological Study ◽  
Shoot Meristem ◽  
Vascular Bundles ◽  
Histological Studies ◽  
Meristematic Cells ◽  
Meristem Development

<div><table cellspacing="0" cellpadding="0" align="center"><tbody><tr><td align="left" valign="top"><p>Since coconut is   one of the most recalcitrant species to generate <em>in vitro</em>, it is   necessary to study in detail about the cellular changes that occur during   somatic embryogenesis to enhance our knowledge about this phenomenon. In the   present study, coconut plumular tissues, the shoot meristem including leaf   primordia, were used as explants for <em>in vitro </em>regeneration studies.   Histological studies were carried out in different stages of plumule culture.   No noticeable growth was observed in 15 days old cultures. After 30 days,   meristematic cells could be identified. Abundance of meristematic cells,   foremost to the development of callus structures, was observed after 45 days.   After 75 days, globular friable calli were formed and histological studies   revealed the presence of meristematic centers which eventually formed somatic   embryos. The histological study of matured somatic embryos formed after 120   days of callus initiation showed a clear meristematic zone of parenchyma   cells, surrounded by vascular bundles. Histological studies, carried out for   certain abnormalities like compact calli, abnormal somatic embryoids with   rudimentary shoots and multiplied roots, revealed the presence of intact   cotyledonary leaves which seemed to inhibit the apical meristem development   of somatic embryoids. The presence of vascular bundles in the early stages of   callus formation might lead to the direct formation of meristemoids. These   results could aid future studies leading to enhanced control of the somatic   embryogenic process and greater efficiency of somatic embryo and plantlet   formation in coconut.</p></td></tr></tbody></table></div>

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