A successful application of the embryo rescue technique as a model for studying crosses between Salix viminalis and Populus species

2011 ◽  
Vol 59 (4) ◽  
pp. 382 ◽  
Author(s):  
Agnieszka Bagniewska-Zadworna ◽  
Maciej Zenkteler ◽  
Elzbieta Zenkteler ◽  
Maria K. Wojciechowicz ◽  
Abdelali Barakat ◽  
...  
Keyword(s):  
Embryo Rescue ◽  
Female Parent ◽  
Botanical Garden ◽  
Intergeneric Hybrids ◽  
Salix Viminalis ◽  
Putative Hybrid ◽  
Ms Medium ◽  
Hybrid Nature ◽  
Rescue Technique

Embryos and plants from the crosses Salix viminalis L. as the female parent and Populus alba L., P. violascens Dode or P. tremula L. as the male parent were obtained by in vitro embryo rescue technique. F1 intergeneric progeny were obtained by overcoming post-zygotic barriers caused by the deficiency of endosperm as nutritive tissue during the first stage of embryo development. Ovules containing immature heart-stage embryos as well as early cotyledonary embryos were isolated and rescued in modified 1/2 MS medium supplemented with 3% sucrose to ensure maturity. Seedlings were cultured under in vitro conditions in 1/2 MS medium that contained 0.2 mg l–1 naphtaleneacetic acid and plantlets after rooting were transferred first into pots and subsequently to the experimental field in the Botanical Garden in Poznań, Poland. After 2 years, the putative hybrid nature of the plants was checked by analysing morphological characters and molecular markers. Scanning electron microscopy, flow cytometry and random amplification of polymorphic DNA screening of individuals confirmed the hybrid nature of the S. viminalis × P. alba and S. viminalis × P. violascens progeny and of 25% of the analysed plants from the cross S. viminalis × P. tremula. This study showed the feasibility of overcoming pre- and post-fertilisation barriers to achieve intergeneric hybrids between species from two genera of Salicaceae. The in vitro system to produce hybrids with characters of both parents could be of great importance for increasing biomass production. This study also opens new opportunities to improve other traits in trees, such as resistance to pathogens inherited from one parent.

scholarly journals (240) Cytological Analyses of BC1 Hybrids Derived from IMR 2n Gamete of Interspecific Hybrid Between L. auratum and L. henryi

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1040D-1041 ◽  
Author(s):  
Mi-Young Chung ◽  
Jae-Dong Chung ◽  
Jaap Van Tuyl ◽  
Ki-Byung Lim
Keyword(s):  
Embryo Rescue ◽  
Female Parent ◽  
Genome Composition ◽  
Germination Ability ◽  
Fertile Pollen ◽  
Rescue Technique ◽  
2N Gamete ◽  
Chromosome Segments ◽  
Morphological Phenotype

Intergenomic F1 hybrids between L. auratum and L. henryi were made and backcrossed to Oriental hybrids to produce BC1 progenies. The F1 intergenomic hybrids produced a relevant frequency of 2n-gamete and fertile pollen. Analyses of pollen size, viability, and germination ability were checked under microscopic observation. GISH analysis confirmed that F1 intergenomic hybrids possess a set of chromosomes from both parents and showed an intermediate morphological phenotype. Twenty BC1 plants were generated by in vitro embryo rescue technique, and analyzed for genome composition by GISH. All plants were triploid, showing 12 from female parent (2× Oriental) and 24 from male (2× F1 hybrid). Based on the GISH analyses, 19 BC1 plants were derived from FDR 2n-gamete and only one plant was derived from IMR 2n-gamete of F1 hybrid. The genome composition of 19 BC1 plants from FDR 2n-gamete possess 12 Orientals + 12 L. auratum + 12 L. henryi chromosomes with some extent of homoeologous recombination between L. auratum and L. henryi. However, one plant from IMR 2n-gamete origin contains an odd number of parental chromosomes from F1 hybrid, showing 12 Oriental + 14 L. auratum + 10 L. henryi chromosomes. In this case, two L. auratum chromosomes recombinant with L. henryi chromosome segments were added and L. henryi counterpart chromosomes were deleted, respectively.

scholarly journals Enhanced Seed Germination of Ilex dumosa R. (Aquifoliaceae) through In Vitro Culture of Cut Pyrenes

HortScience ◽  
2011 ◽  
Vol 46 (2) ◽  
pp. 278-281 ◽  
Author(s):  
Natalia R. Dolce ◽  
Luis A. Mroginski ◽  
Hebe Y. Rey
Keyword(s):  
In Vitro Culture ◽  
Embryo Rescue ◽  
Germination Percentage ◽  
Effective Technique ◽  
Cutting Surface ◽  
Rescue Technique ◽  
Growth Room ◽  
Scalpel Blade ◽  
Culture Protocol

An in vitro culture protocol was developed that increased the germination percentage and decreased the lag time to germination for Ilex dumosa R. pyrenes as a tool for replacing the laborious task of embryo rescue technique. This method involves transversely cutting surface-sterilized pyrenes with a scalpel blade, then placing the micropylar one-third end with the rudimentary embryo (≈0.25 mm long) on solidified (agar 0.65%) quarter-strength salts and vitamins of Murashige and Skoog, 1962 medium with 3% sucrose, and incubating in a growth room at 27 ± 2 °C with a 14-h photoperiod (116 μmol·m−2·s−1). Most of the cut pyrenes (greater than 50%) germinated within the first month after inoculation and achieved maximum germination (≈70%) in 2 months compared with whole pyrenes, which began to germinate 3 months after sowing and required more than 8 months for maximum germination (37%). Moreover, the germination percentage of cut pyrenes was significantly higher than the germination of isolated embryos (34%). Thus, the cut pyrenes culture is a simpler and more effective technique than embryo rescue. Easily, on average, a trained operator is able to culture ≈1000 cut pyrenes per day instead of ≈100 isolated embryos.

scholarly journals Embryo rescue from interspecific crosses in apple rootstocks

2006 ◽  
Vol 41 (6) ◽  
pp. 969-973 ◽  
Author(s):  
Adriana Cibele de Mesquita Dantas ◽  
José Itamar Boneti ◽  
Rubens Onofre Nodari ◽  
Miguel Pedro Guerra
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Embryo Rescue ◽  
Casein Hydrolysate ◽  
Female Parent ◽  
Interspecific Crosses ◽  
Open Pollination ◽  
Apple Rootstocks ◽  
Hand Pollination

The objetive of this work was to rescue immature embryos of apple rootstocks Malus prunifolia (Marubakaido) and Malus pumila (M9) after 40-60 days of pollination and to put them into MS culture media supplemented with agar (6 g L-1) and casein hydrolysate (500 mg L-1). Embryos originated from interspecific crosses and open pollination showed differences in the in vitro responses, depending on the female parent, the developmental stage of the embryo, and the culture medium composition. Embryos of the M. pumila rootstock, rescued within 40 days after pollination and put in culture medium supplemented with indolacetic acid (IAA), gibberellic acid (GA3), kinetin and maltose, resulted in a normal development of plantlets. However, embryos originating from hand-pollination, cultivated in medium supplemented with 14 µM IAA, 5 µM kinetin and 1.5 µM Ga3 (MS1), mainly those of M. prunifolia x M. pumila, showed a high percentage of rusted embryos (96.2%). Embryos from open pollination of M. prunifolia and M. pumila formed calluses. It was possible to identify the influence of the female parent by the enhanced development of M. pumila shoots derived from open or hand-pollination. The crossing of responsive species and the use of the technique of embryo culture provided a rapid and uniform germination and, consequently, the development of fully normal seedlings.

A study on genomic relationships of Agropyron trachycaulum with Elymus scabriglumis, E. innovatus, and Hordeum procerum

Genome ◽  
1988 ◽  
Vol 30 (4) ◽  
pp. 525-528 ◽  
Author(s):  
P. K. Gupta ◽  
H. S. Balyan ◽  
George Fedak
Keyword(s):  
Chiasma Frequency ◽  
Embryo Rescue ◽  
Intergeneric Hybrids ◽  
Genomic Constitution ◽  
Rescue Technique ◽  
Genomic Relationships ◽  
Chromosome Associations ◽  
Synthetic Hybrids ◽  
The Mean ◽  
Agropyron Trachycaulum

Synthetic hybrids of Agropyron trachycaulum (2n = 4x = 28) with Elymus scabriglumis (2n = 6x = 42), E. innovatus (2n = 4x = 28), and Hordeum procerum (2n = 6x = 42) were produced through the embryo rescue technique, at relative frequencies of 7.1, 4.2, and 1.8%, respectively, of pollinated florets. The mean chromosome associations in two hybrids with E. scabriglumis was 11.71II + 1.47III + 0.15IV (mean chiasma frequency = 22.75), in one hybrid with H. procerum was 7.27II + 0.63III + 0.09IV (mean chiasma frequency = 10.20) and in a hybrid with E. innovatus was 2.8II (2.19 chiasma frequency). The data suggested two common genomes (S and H) with E. scabriglumis, one common genome (H) with H. procerum, and no common genome with E. innovatus. The evidence obtained suggested a SSHHYY genomic constitution for E. scabriglumis and two genomes different from SH for E. innovatus. The evidence also suggested that in H. procerum there may not be two related genomes present (derived from H. jubatum) as speculated in some earlier studies.Key words: intergeneric hybrids, Agropyron, Elymus, Hordeum, chromosome pairing.

A Sorghum bicolor × S. macrospermum hybrid recovered by embryo rescue and culture

2005 ◽  
Vol 53 (6) ◽  
pp. 579 ◽  
Author(s):  
H. James Price ◽  
George L. Hodnett ◽  
Byron L. Burson ◽  
Sally L. Dillon ◽  
William L. Rooney
Keyword(s):  
Nuclear Dna ◽  
Embryo Rescue ◽  
Nuclear Dna Content ◽  
Female Parent ◽  
Male Sterile ◽  
Leaf Pubescence ◽  
Culture Techniques ◽  
Close Phylogenetic Relationship ◽  
Improvement Programs

Although exotic germplasm is extensively used in sorghum improvement programs, Sorghum species classified in sections other than Eu-sorghum have not been utilised as germplasm because of strong reproductive barriers involving pollen–pistil incompatibilities. S. macrospermum is of particular interest to sorghum breeders because of its close phylogenetic relationship and cytogenetic similarities to S. bicolor and its resistance to important sorghum pests and pathogens, such as sorghum midge and sorghum downy mildew. A vegetatively vigorous interspecific hybrid was obtained from a cross between a cytoplasmic male-sterile S. bicolor plant and S. macrospermum by using embryo rescue and in vitro culture techniques. The hybrid was morphologically intermediate to S. bicolor and S. macrospermum in leaf width, leaf pubescence, plant height, inflorescence morphology, chromosome number and nuclear DNA content. It was male-sterile like its ATx623 parent. The hybrid produced no offspring when used as the female parent in a backcross with S. bicolor. This is the first confirmed hybrid between S. bicolor and S. macrospermum, and to our knowledge, it is the first reported hybrid between S. bicolor and any Sorghum species outside the Eu-sorghum section.

Optimum age of siliques for rescue of hybrid embryos from crosses between Brassica oleracea, B. rapa and B. carinata

2004 ◽  
Vol 84 (4) ◽  
pp. 965-969 ◽  
Author(s):  
M. H. Rahman
Keyword(s):  
Survival Rate ◽  
Embryo Rescue ◽  
Interspecific Cross ◽  
Female Parent ◽  
Maximum Efficiency ◽  
Interspecific Crosses ◽  
Seed Color ◽  
Female Embryo ◽  
Genome Species ◽  
Rescue Technique

Interspecific hybrids were produced from crosses involving Brassica rapa var. Yellow Sarson (AA), Canadian B. rapa (AA) cultivars, B. oleracea var. alboglabra (CC), B. oleracea var. italica (CC), rapidcycling B. oleracea (CC) and B. carinata (BBCC) to introgress the yellow seed color or self-incompatibility genes into B. napus. Embryo rescue technique was employed for this purpose. In interspecific crosses where a CC genome species was used as the female parent, a significantly lower number of ovules were fertilized compared to the crosses where the BBCC genome species was female. Embryo growth and development were slower in the crosses where the CC genome species was female than in the crosses where BBCC genome species was female. The efficiency of the embryo rescue technique from 16 to 40 d after pollination (DAP) with 4-d intervals was investigated in the crosses using CC genome species as female. The highest numbers of embryos were rescued between 20 and 28 DAP depending upon the specific cross undertaken. However, the survival rate of the embryos rescued at 20 DAP was very low compared to the embryos rescued at 24 and 28 DAP. The survival rate of the embryos rescued at 32 DAP was generally high, but the number of embryos rescued at this stage was significantly lower than at 24 and 28 DAP. No embryos were obtained at 16 DAP or at 40 DAP. Thus, using the CC genome species as female parent, the maximum efficiency of the embryo rescue technique was achieved when embryos were rescued between 24 and 28 DAP. In the case of crosses using the BBCC genome species as female, rescue of hybrid embryos was successful at 18 and 22 DAP. Key words: Brassica, interspecific cross, embryo rescue, embryo development

scholarly journals In vitro embryo rescue of interspecific hybrids of oil palm (Elaeis oleifera x Elaeis guineensis)

2011 ◽  
Vol 2 (2) ◽  
pp. 1-6 ◽  
Author(s):  
Sergio Augusto Oliveira Alves ◽  
Oriel Filgueira de Lemos ◽  
Benedito Gomes dos Santos Filho ◽  
André Luís Lopes da Silva
Keyword(s):  
Oil Palm ◽  
Activated Charcoal ◽  
Interspecific Hybrids ◽  
Elaeis Guineensis ◽  
Germination Rate ◽  
Embryo Rescue ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Elaeis Oleifera

The oil palm (Elaeis guineennsis Jacq.) is the plant with major oil yield in tropical areas over the world. Pará state is the major producer of Brazil. However, the production of resistant hybrids (Elaeis oleifera x Elaeis guinennsis) to several diseases is necessary, in spite of less productive, is a viable alternative for areas where occur more aggressive diseases. The aim of this research was to rescue in vitro embryos of four varieties of interspecific hybrids of oil palm (Elaeis oleifera x Elaeis guineensis) in different media. Four varieties were tested: (1) Cj-2141; (2) CI-2061; (3) Cj-502 and (4) Cj-494. Four media were tested: (T1) half strength MS medium (½ MS), (T2) MS, (T3) MS + 0.5mg L-1 1-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP) and (T4) ½ MS+ 0.5mg.L-1 NAA and BAP. All media were supplemented with 0.17g.L-1 NaH2PO4, activated charcoal (0.25%), sucrose (3%) and solidified with phytagel (0.2%). The most germination rate of interspecific hybrids embryos was obtained on ½ MS medium, supplemented with 0.17g.L-1 NaH2PO4, activated charcoal (0.25%), sucrose (3%) and solidified with phytagel (0.2%). The variety CI-2061 presents the best capacity to develop normal seedlings.

Cytological analysis of hybrid embryos of intergeneric crosses between Salix viminalis and Populus species

2010 ◽  
Vol 58 (1) ◽  
pp. 42 ◽  
Author(s):  
Agnieszka Bagniewska-Zadworna ◽  
Maria K. Wojciechowicz ◽  
Maciej Zenkteler ◽  
Stanisław Jeżowski ◽  
Elżbieta Zenkteler
Keyword(s):  
Laser Scanning ◽  
Populus Trichocarpa ◽  
Embryo Rescue ◽  
Laser Scanning Microscopy ◽  
Salix Viminalis ◽  
Confocal Laser ◽  
Hybrid Plants ◽  
Populus Species ◽  
Intergeneric Hybridisation

Intergeneric hybridisation between Salix viminalis L. as the female and four Populus species (Populus trichocarpa, P. tremula, P. × canadensis and P. simonii) as male pollen donors was performed by in vitro stigma pollination. To overcome postzygotic barriers, transfer of hybrid embryos to new medium is necessary. We carried out detailed ultrastructural analyses to establish: (i) at which stage of embryo development the first signs of programmed cell death (PCD) could be detected; and (ii) at which stage the lack of serious or irreversible changes guaranteed that advanced development of hybrid plants could occur after embryo rescue. Transmission electron microscopy and confocal laser scanning microscopy revealed the presence of both developing and degenerating embryos. Developing globular, heart-shaped, and early cotyledonary embryos contained cells of correct ultrastructure. The only sign of intergeneric hybridisation was a delay in development for a few days, in comparison with control embryos. The earliest indicators of embryo degeneration were noted at 9 days after pollination (DAP). The most common indicators were excessive embryo vacuolisation, which was characterised by a large number of vesicles and formation of small vacuoles, as well as enlarged central vacuoles. Extended plastid thylakoids, folding of the cell wall, and autophagosomes were observed. Our detailed investigation of PCD in hybrid embryos enabled us to conclude that the embryo rescue technique was most effective in intergeneric willow × poplar crosses if applied between 9 and 16 DAP.

Progress in backcrossing between Carica papaya × Vasconcellea quercifolia intergeneric hybrids and C. papaya

2006 ◽  
Vol 46 (3) ◽  
pp. 419 ◽  
Author(s):  
R. A. Drew ◽  
S. V. Siar ◽  
C. M. O'Brien ◽  
A. G. C. Sajise
Keyword(s):  
Pollen Fertility ◽  
Carica Papaya ◽  
Embryo Rescue ◽  
The Philippines ◽  
Intergeneric Hybrids ◽  
Tropical Fruit ◽  
Hybrid Plants ◽  
Large Numbers ◽  
Backcross Populations

Papaya is a major tropical fruit crop worldwide, however, all genotypes are susceptible to Papaya ringspot virus type P (PRSV-P). Protocols have been developed to produce large numbers of intergeneric hybrids between Carica papaya L. and Vasconcellea quercifolia, which is PRSV-P resistant. Although pollen fertility of these hybrids was low, backcross populations were produced in both Australia and the Philippines when papaya flowers were pollinated with pollen from 11 hybrid plants that had been identified as having some pollen fertility and were PRSV-P resistant. Plantlets were produced after embryo rescue and culture in vitro. Second backcross generations (BC2) were easier to produce than first backcross (BC1) generations as pollen fertility was >80% in male BC1 plants. Variation in time to PRSV-P symptom expression and severity of virus symptoms in the backcross generations was observed in both countries. Commercially acceptable fruit were produced in the BC2 generation. The results demonstrate that efforts in wide hybridisation to transfer PRSV-P resistance to C. papaya, are better directed towards crosses between C. papaya and V. quercifolia than with other Vasconcellea species.

scholarly journals Polyploidization Facilitates BiotechnologicalIn VitroTechniques in the GenusCucumis

2010 ◽  
Vol 2010 ◽  
pp. 1-8
Author(s):  
Dagmar Skálová ◽  
Vladan Ondřej ◽  
Ivana Doležalová ◽  
Božena Navrátilová ◽  
Aleš Lebeda
Keyword(s):  
Ploidy Level ◽  
Embryo Rescue ◽  
Mesophyll Protoplast ◽  
Interspecific Crosses ◽  
Putative Hybrid ◽  
Genetic Barriers ◽  
Hybrid Character ◽  
Positive Effect

Prezygotic interspecific crossability barrier in the genusCucumisis related to the ploidy level of the species (cucumber (C. sativus), ; muskmelon (C. melo) and wildCucumisspecies, ). Polyploidization of maternal plants helps hybridization among otherCucumisspecies by overcoming prezygotic genetic barriers. The main objective of this paper is to compare the results of several methods supporting interspecific crosses in cucumber without and with polyploidization (comparison between diploid (2x) and mixoploid (2x/4x) cucumber maternal plants). Mixoploid plants were obtained afterin vivoandin vitropolyploidization by colchicine and oryzalin. Ploidy level was estimated by flow cytometry. Embryo rescue,in vitropollination, and isolation of mesophyll protoplast were tested and compared. Positive effect of polyploidization was observed during all experiments presented by higher regeneration capacity of cultivated mixoploid cucumber embryos, ovules, and protoplasts. Nevertheless, the hybrid character of putative hybrid accessions obtained after crossin vivoandin vitropollination was not confirmed.

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