In vitro control of fasciation of nucellar embryos in the proliferating embryogenic cultures of a monoembryonic mango variety Chausa

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.

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Somaclonal Variation During Picea abies and P. omorika Somatic Embryogenesis and Cryopreservation

Acta Biologica Cracoviensia s Botanica ◽

10.1515/abcsb-2017-0003 ◽

2017 ◽

Vol 59 (1) ◽

pp. 93-103 ◽

Cited By ~ 2

Author(s):

Teresa Hazubska-Przybył ◽

Monika Dering

Keyword(s):

Somatic Embryogenesis ◽

Picea Abies ◽

Somaclonal Variation ◽

Genetic Stability ◽

Somatic Embryos ◽

Plant Material ◽

Stress Factors ◽

Embryogenic Cultures ◽

Embryogenic Lines

AbstractEmbryogenic cultures of plants are exposed to various stress factors bothin vitroand during cryostorage. In order to safely include the plant material obtained by somatic embryogenesis in combination with cryopreservation for breeding programs, it is necessary to monitor its genetic stability. The aim of the present study was the assessment of somaclonal variation in plant material obtained from embryogenic cultures ofPicea abies(L.) Karst. andP. omorika(Pančić) Purk. maintainedin vitroor stored in liquid nitrogen by the pregrowth-dehydration method. The analysis of genetic conformity with using microsatellite markers was performed on cotyledonary somatic embryos (CSE), germinating somatic embryos (GSE) and somatic seedlings (SS), obtained from tissues maintainedin vitroor from recovered embryogenic tissues (ETc) and CSE obtained after cryopreservation. The analysis revealed changes in the DNA of somatic embryogenesis-derived plant material of bothPiceaspp. They were found in plant material from 8 out of 10 tested embryogenic lines ofP. abiesand in 10 out of 19 embryogenic lines ofP. omorikaafterin vitroculture. Changes were also detected in plant material obtained after cryopreservation. Somaclonal variation was observed in ETc and CSE ofP. omorikaand at ETv stage ofP. abies. However, most of the changes were induced at the stage of somatic embryogenesis initiation. These results confirm the need for monitoring the genetic stability of plants obtained by somatic embryogenesis and after cryopreservation for both spruce species.

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Effect of carbon source on morphology and histodifferentiation of Araucaria angustifolia embryogenic cultures

Brazilian Archives of Biology and Technology ◽

10.1590/s1516-89132005000800005 ◽

2005 ◽

Vol 48 (6) ◽

pp. 895-903 ◽

Cited By ~ 36

Author(s):

Neusa Steiner ◽

Felipe do Nascimento Vieira ◽

Sara Maldonado ◽

Miguel Pedro Guerra

Keyword(s):

Carbon Source ◽

Somatic Embryos ◽

Zygotic Embryos ◽

Araucaria Angustifolia ◽

Embryogenic Cultures ◽

Important Species ◽

Induction Rate

The aim of the present work was to establish in vitro conditions for the induction, stabilization and proliferation of embryogenic cultures of A. angustifolia. Pre-cotyledonary staged zygotic embryos inoculated BM medium supplemented with 5 µM 2,4- D, 2 µM BAP and Kin, and 3% maltose or sucrose resulted in 66.7% induction rate. The rate of induction of embryogenic cultures was affected by the carbon source, as well the multiplication and morphology of the embryogenic cultures. Embryogenic cultures maintained in BM medium with maltose presented bipolar morphology. Globular somatic embryos were obtained BM medium with 9% (PEG) and (9%) maltose. These results could establish an in vitro regenerative protocol towards the conservation and improvement of this important species.

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Use of Plant Antimicrobial Peptides in in vitro Embryogenic Cultures of Larix sibirica

Biology Bulletin ◽

10.1134/s1062359020030097 ◽

2020 ◽

Vol 47 (3) ◽

pp. 225-236

Author(s):

I. N. Tretyakova ◽

E. A. Rogozhin ◽

M. E. Pak ◽

I. A. Petukhova ◽

A. S. Shuklina ◽

...

Keyword(s):

Antimicrobial Peptides ◽

Larix Sibirica ◽

Embryogenic Cultures

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In vitro conservation of oil palm somatic embryos for 20 years on a hormone-free culture medium: characteristics of the embryogenic cultures, derived plantlets and adult palms

Plant Cell Reports ◽

10.1007/s00299-009-0787-y ◽

2009 ◽

Vol 29 (1) ◽

pp. 1-13 ◽

Cited By ~ 29

Author(s):

K. Eugene Konan ◽

Tristan Durand-Gasselin ◽

Y. Justin Kouadio ◽

Albert Flori ◽

Alain Rival ◽

...

Keyword(s):

Oil Palm ◽

Somatic Embryos ◽

Culture Medium ◽

In Vitro Conservation ◽

Embryogenic Cultures

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In vitro conservation of embryogenic cultures of date palm using osmotic mediated growth agents

Journal of Genetic Engineering and Biotechnology ◽

10.1016/j.jgeb.2016.08.004 ◽

2016 ◽

Vol 14 (2) ◽

pp. 363-370 ◽

Cited By ~ 8

Author(s):

M.K. El-Bahr ◽

A. Abd EL-Hamid ◽

M.A. Matter ◽

A. Shaltout ◽

S.A. Bekheet ◽

...

Keyword(s):

Date Palm ◽

In Vitro Conservation ◽

Embryogenic Cultures

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Micropropagation Cultures for Genetic Transformation of Grapevine

HortScience ◽

10.21273/hortsci.41.4.972c ◽

2006 ◽

Vol 41 (4) ◽

pp. 972C-972

Author(s):

Manjul Dutt ◽

Dennis J. Gray ◽

Zhijian T. Li ◽

Sadanand Dhekney ◽

Marilyn M. Van Aman

Keyword(s):

Genetic Transformation ◽

Fluorescent Protein ◽

Marker Gene ◽

In Vitro Cultures ◽

35S Promoter ◽

Nptii Gene ◽

Embryogenic Cultures ◽

Wide Range ◽

Over Time

A major drawback to the use of embryogenic cultures for transformation of grapevine is that their ability to undergo genetic transformation is cultivar-dependent. Also, depending on cultivar, embryogenic cultures are difficult to impossible to maintain over time, reducing their utility for use in genetic transformation. An alternative to the use of embryogenic cultures for transformation of grapevine is the use of micropropagation cultures, which are easier to initiate from a wide range of grapevine cultivars and can be maintained over time without loss of function. Vitis vinifera `Thompson Seedless' was used as a model for genetic transformation using micropropagation cultures. In vitro cultures were initiated from apical meristems of actively growing vines and maintained in C2D medium containing 4 μM of 6-benzylaminopurine (C2D4B). Shoot tips and nodes were collected from proliferating in vitro cultures for transformation studies. A variety of wounding techniques, including nicking, sonication, and fragmenting of meristematic tissues was employed in order to enable Agrobacterium infection. We used a construct containing a bidirectional 35S promoter complex with a marker gene composed of a bifunctional fusion between an enhanced green fluorescent protein (EGFP) gene and a neomycin phosphotransferase (NPTII) gene in one direction and a hybrid lytic peptide gene in the other. Transgenic shoots growing in C2D4B medium containing 200 mg·L-1 each of carbenicillin and cefotaxime and 20 mg·L-1 of kanamycin were selected based on GFP fluorescence. Transgenic shoots were rooted and transferred to a greenhouse. To date, 18 transgenic lines have been generated. Details on the transformation procedure will be discussed.

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Optimizing Initiation and Maintenance of Vitis Embryogenic Cultures

HortScience ◽

10.21273/hortsci.44.5.1400 ◽

2009 ◽

Vol 44 (5) ◽

pp. 1400-1406 ◽

Cited By ~ 28

Author(s):

Sadanand A. Dhekney ◽

Zhijian T. Li ◽

Michael E. Compton ◽

Dennis J. Gray

Keyword(s):

Somatic Embryogenesis ◽

Culture Media ◽

Medium Composition ◽

Embryogenic Competence ◽

Embryogenic Cultures ◽

Vitis Riparia ◽

Vitis Rupestris ◽

In Vitro Micropropagation ◽

Embryogenic Response

Stamens and pistils from mature grapevines and leaves from in vitro micropropagation cultures were used to optimize parameters influencing somatic embryogenesis in Vitis. Embryogenic competence was dependent on species/variety, explant type and developmental stage, medium composition, and growth regulator concentration. Of varieties evaluated, a greater number produced embryogenic cultures from stamens and pistils (26) compared with leaves (six). Among the different stamen and pistil stages, Stage II and III explants produced the maximum embryogenic response regardless of genotype and medium composition. Of seven culture media tested, the highest embryogenic response was recorded from varieties cultured on MSI (18) and PIV (16) media. Experiments annually repeated over 3 to 10 years demonstrated reproducible results. Highly reliable protocols for somatic embryogenesis were obtained for 29 Vitis species and varieties, including 18 Vitis vinifera varieties, Vitis riparia, Vitis rupestris, Vitis champinii, and eight Vitis hybrids. Embryogenic cultures were maintained on X6 medium for a period of 6 months to 2 years depending on the variety and used in studies involving genetic transformation and transgenic plant regeneration.

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Shoot Micropropagation in Vitis as a Source of Leaf-derived Embryogenic Cultures

HortScience ◽

10.21273/hortsci.30.4.757f ◽

1995 ◽

Vol 30 (4) ◽

pp. 757F-757

Author(s):

M. Meyerson ◽

C.M. Benton ◽

D.J. Gray

Keyword(s):

Cabernet Sauvignon ◽

Embryogenic Cultures ◽

Du Bois ◽

In Vitro Micropropagation ◽

Shoot Number ◽

Significant Difference ◽

Micropropagated Plants ◽

Sterile Leaf

Micropropagation of Vitis bourquiniana Lenoir `Black Spanish', V. champini Planchon `Dog Ridge', Vitis hybrids (`Blanc du Bois', `Himrod', and `Niagara Seedless'), V. rotundifolia Michx. (`Carlos' and `Dixie'), and V. vinifera L. (`Autumn Seedless', `Cabernet Sauvignon', `Carignane', `French Colombard', `Ruby Cabernet', and `Tokay') was accomplished. Shoot tips taken from micropropagated plants in long-term culture were inoculated onto solidified C2D medium containing 5 μM benzyladenine. Culture times consisting of either one or two 4-week cycles were compared for effect on shoot number. A range of response among cultivars tested was noted. The best-responding variety was V. champini `Dog Ridge', with 5.8 shoots per apex. All other varieties were less prolific. When shoot micropropagation from nodal explants and apices was compared, so significant difference was noted. In vitro micropropagation offers rapid clonal production of grape and is a source of sterile leaf explant material for embryogenic cultures, which, in turn, are useful target for genetic transformation.

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Somatic embryogenesis and plant regeneration from immature embryos of Carica papaya × Carica cauliflora cultured in vitro

Canadian Journal of Botany ◽

10.1139/b91-240 ◽

1991 ◽

Vol 69 (9) ◽

pp. 1913-1918 ◽

Cited By ~ 27

Author(s):

M. H. Chen ◽

C. C. Chen ◽

D. N. Wang ◽

F. C. Chen

Keyword(s):

Somatic Embryogenesis ◽

Plant Regeneration ◽

Somatic Embryos ◽

Interspecific Hybrids ◽

Carica Papaya ◽

Immature Embryos ◽

Embryogenic Cultures ◽

Regenerated Plants ◽

Hybrid Embryo

Somatic embryos were induced directly on immature embryos of Carica papaya × Carica cauliflora hybrids cultured on modified Murashige and Skoog's medium. When transferred to medium supplemented with abscisic acid, individual somatic embryos proliferated numerous daughter embryos through repeated embryogenesis. Light microscopic study of the repeatedly embryogenic cultures showed that daughter embryos arose from single superficial cells of parent embryos. Plant regeneration occurred following transfer of somatic embryos to medium devoid of plant growth regulators. Regenerated plants were intermediate between C. papaya and C. cauliflora in several morphological respects and showed isozyme patterns specific to both species as well as some new bands, indicating that they are indeed interspecific hybrids. Key words: Carica, interspecific hybrid, embryo culture, somatic embryogenesis.

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