Transformation of an Australian Variety of Carica papaya Using Microprojectile Bombardment

1996 ◽  
Vol 23 (6) ◽  
pp. 679 ◽  
Author(s):  
RE Mahon ◽  
MF Bateson ◽  
DA Chamberlain ◽  
CM Higgins ◽  
RA Drew ◽  
...  
Keyword(s):  
Transient Expression ◽  
Somatic Embryos ◽  
Carica Papaya ◽  
Day Treatment ◽  
Microprojectile Bombardment ◽  
Target Tissue ◽  
System Transformation ◽  
Uida Reporter Gene ◽  
Osmotic Treatment ◽  
Regenerated Plantlets

We have developed a method for the stable transformation and regeneration of a dioecious Australian cultivar of Carica papaya (papaw or papaya) by microprojectile bombardment. This method was developed after investigation of both zygotic and somatic embryos as target tissue and optimisation of a number of parameters using transient expression of the uidA reporter gene. The tissue culture regime prior to bombardment was critical in optimisation of transformation. Factors such as age of embryos and various treatments prior to bombardment, increased transient expression by up to 22-fold. Highest uidA transient expression results were obtained when somatic embryos 3 weeks since last subculture on solid medium were given a 3 day treatment in liquid medium, and a 2 h osmotic treatment pre- and post-bombardment. Stably transformed plants were obtained 6 months after bombardment using this system. Transformation efficiency was high with two experiments yielding 45% and 37.5% of bombarded plates regenerating plantlets on media containing kanamycin. The presence of both the uidA and aphA genes, (selectable marker) which code for the enzymes β-glucuronidase (GUS), and neomycin phosphotransferase II (NPTII) respectively, was confirmed in regenerated plantlets by Southern hybridisation.

scholarly journals Biolistic-mediated transient gene expression in shoot apical meristems of the prickly-pear (Opuntia ficus-indica)

1999 ◽  
Vol 42 (3) ◽  
pp. 299-302 ◽  
Author(s):  
Romulo Marino Llamoca-Zárate ◽  
Luiz Ferreira Aguiar Ponte ◽  
Joerg Landsmann ◽  
Francisco de Assis Paiva Campos
Keyword(s):  
Gene Expression ◽  
Transient Expression ◽  
Transient Gene Expression ◽  
Dna Delivery ◽  
Prickly Pear ◽  
Target Tissue ◽  
Gus Gene ◽  
Transformation System ◽  
Opuntia Ficus Indica ◽  
Apical Meristems

We have demonstrated the transient expression of the GUS gene in cells of the meristematic apical dome of Opuntia ficus-indica. DNA delivery into the cells was achieved using a biolistic PDS-1000He instrument from Bio-Rad Laboratories. The transforming DNA was coated in tungsten particles with diameter of 1.3 m m and the distance between the flying disk and the target tissue was 7.5cm and the shooting pressure was adjusted to 1200 psi. This is the first demonstration that the biolistic transformation system can be used to express a transgene in a member of the Cactaceae.

Transient expression of marker genes in immature zygotic embryos of spring wheat (Triticum aestivum) through microprojectile bombardment

Genome ◽  
1991 ◽  
Vol 34 (3) ◽  
pp. 453-460 ◽  
Author(s):  
R. N. Chibbar ◽  
K. K. Kartha ◽  
N. Leung ◽  
J. Qureshi ◽  
K. Caswell
Keyword(s):  
Triticum Aestivum ◽  
Transient Expression ◽  
Microprojectile Bombardment ◽  
Triticum Aestivum L ◽  
Cauliflower Mosaic Virus ◽  
Expression Vectors ◽  
Marker Genes ◽  
Zygotic Embryos ◽  
Coding Region ◽  
Immature Zygotic Embryos

Transient expression of marker genes (cat and uidA) delivered by the Biolistics microprojectile bombardment technique has been detected in immature zygotic embryos of wheat (Triticum aestivum L.). The DNA expression vectors that gave maximal expression of both cat (pCaMVI1CN) and uidA (pCaMVI1GusN) genes had an alcohol dehydrogenase (Adh1) intron 1 cloned in between the cauliflower mosaic virus (CaMV35S) promoter and the coding region of the gene. Detection of chloramphenicol acetyltransferase (CAT) activity in response to cat gene was complicated by the presence of an inhibitor of CAT activity as well as an endogenous CAT-Iike activity. The results of enzymatic assays were confirmed by an ELISA technique using CAT-specific antibodies, whereas the β-glucuronidase (GUS) activity following the introduction of the uidA gene was confirmed by both histochemical and fluorometric techniques.Key words: cereal transformation, gene expression, ELISA, wheat.

scholarly journals Partial Organogenesis and Somatic Embryogenesis from Petiole Explants of Field-grown Papaya (Carica papaya L.)

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 629g-630 ◽  
Author(s):  
S. Jayasankar ◽  
U.L Yadava
Keyword(s):  
Somatic Embryogenesis ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Carica Papaya ◽  
Vascular Tissue ◽  
The Other ◽  
Ms Medium ◽  
Free Medium ◽  
Petiole Explants ◽  
Young Leaves

Petiole discs from young leaves of female papaya (L-45) plants were cultured in MS or B5-based media containing 0, 2.25, 4.5, 11.25, and 22.5 μm 2,4-D. Compact embryogenic callus emerged from vascular tissue of petiole discs in about 3 weeks. In MS medium, 66% and 51% explants formed embryogenic callus with 11.25 and 22.5 μm 2,4-D, respectively. On the other hand, 79% explants formed embryogenic callus in B5-based medium with 4.50 μm 2,4-D. However, explants became necrotic in B5-based medium with 22.5 μm 2,4-D. Subculturing callus in auxin-free medium resulted in the development of roots or somatic embryos. Microscopic observations revealed that the roots were produced only by the callus that had retained its continuity with the vascular tissue. This investigation revealed that petioles from field grown papaya plants are potential explants for somatic embryogenesis and 2-week exposure to 2,4-D is adequate for inducing morphogenesis. Additionally, an interaction between 2,4-D and the components in the MS and B5-based media was observed.

scholarly journals Effect of water stress induced by polyethylene glycol 6000 on the somatic embryos conversion into whole plantlets in cocoa (Theobroma cacao L.)

2021 ◽  
Vol 15 (1) ◽  
pp. 12-25
Author(s):  
Tokpapon Eliane Manlé ◽  
Kan Modeste Kouassi ◽  
Brahima André Soumahoro ◽  
Tchoa Koné ◽  
Kouablan Edmond Koffi ◽  
...  
Keyword(s):  
Climate Change ◽  
Polyethylene Glycol ◽  
Water Stress ◽  
Somatic Embryos ◽  
Stem Length ◽  
Improvement Program ◽  
Polyethylene Glycol 6000 ◽  
Regenerated Plantlets ◽  
Theobroma Cacao L

Rainfall scarcity due to climate change is a major constraint that limits cocoa productivity in Côte d'Ivoire. This work aims to regenerate cocoa plants tolerant to water stress using in vitro methods. Staminode and petal explants of the genotypes C1, C9, C14, C15, C16, C18 and C20 were used to produce somatic embryos through two methods. Firstly, somatic embryos were induced under stressfull conditions on media containing different concentrations of polyethylene glycol (PEG) 6000 (0; 25; 50; 75; 100 and 125 g/l) and secondly; under non-stressed conditions. Somatic embryos were placed on a conversion medium in the same stress condition. The number of regenerants decreased with the increase in the concentration of PEG with all genotypes. Only genotypes C1 and C15 regenerated plantlets under water stress conditions. The sensitive genotypes C9, C14, C16, C18 and C20 have not developed plantlets on media containing PEG. The plantlets produced under water deficit conditions exhibited a reduction in stem length and leaves number and an increase in length or offset of the high number of roots. The survival rate of regenerants during acclimatization was higher on the sandsubstrate. The selected genotypes could be used in an improvement program of cocoa production.Keywords: Climate change; plant regeneration; genotype; tolerance; drought; in vitro

scholarly journals Somatic Embryogenesis using Immature Zygotic Embryos from Developing Fruits of Papaya (Carica papaya)

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 783E-783
Author(s):  
S.K. Dhir ◽  
U.L. Yadava
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Carica Papaya ◽  
Induction Medium ◽  
Zygotic Embryos ◽  
Synthetic Seed ◽  
Factors Affecting ◽  
Secondary Embryos ◽  
Potential Use

An efficient protocol has been developed for the in vitro multiplication of papaya (Carica papaya L.) through somatic embryogenesis utilizing immature zgotic embryos. Somatic embryos were initiated on MS basel media supplemented with 5 mg·liter–1 2,4-D, 400 mg·liter–1 glutamine, and 6% sucrose. After culturing for 2 months, 65% of the explants became highly embryogenic. Each explant produced 50 to 80 embryos in 4 months on culture induction medium. Frequency of embryogenesis was increased (75 to 150 somatic embryos on 80% explants) upon supplementing medium with 4% maltose as a carbon source and 100 mg·liter–1 L-asparagine. The embryogenic callus appeared yellow and embryos at different stages of development were well-organized. On regular subculturing, these cultures continued to produce secondary embryos. Following their transfer to the hormone-free medium supplemented with 4% maltose, these embryos germinated. The somatic embryogenesis system is rapid, repetitive, and highly proliferative. Thus, this system may have a potential use in the development of synthetic seed and transgenic papaya plants. Details of important factors affecting somatic embryogenesis will be discussed.

scholarly journals Plant regeneration through somatic embryogenesis from calli derived from leaf bases of Laurus nobilis L. (Lauraceae)

2015 ◽  
Vol 24 (2) ◽  
pp. 213-221 ◽  
Author(s):  
Ahmad H Al Gabbiesh ◽  
M Ghabeish ◽  
I H ◽  
M Kleinwächter ◽  
D Selmar
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Root Formation ◽  
Callus Formation ◽  
Germplasm Conservation ◽  
Laurus Nobilis ◽  
The Media ◽  
Regenerated Plantlets ◽  
Culture Technology

Somatic embryogenesis was induced in embryo culture on half MS medium supplemented with NAA (8 mg/l) as the sole plant growth regulator after incubation of the media in the refrigerator at 4°C for two weeks to promote callus induction and somatic embryogenesis in Laurus nobilis. Both embryogenetic calli and somatic embryos were induced in the above selected medium. Embryo growth and development were stimulated by separation of embryos successfully from embryo clusters and transferred onto fresh half MS. Among the selected explants, only leaf bases were found to respond actively to plant regeneration, especially in inducing callus formation and in sustaining faster callus growth. Root formation of regenerated plantlets tended to decrease with time on regeneration media. Overall, 75% of the plantlets derived from the callus survived in the greenhouse; and they all grew to phenotypically normal plants. This procedure will enable the use of regeneration tissue culture technology for germplasm conservation of L. nobilis, a plant of high medicinal and commercial value.Plant Tissue Cult. & Biotech. 24(2): 213-221, 2014 (December)

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