scholarly journals Seed and Embryo Growth in Pod Cultures of Phaseolus vulgaris and P. vulgaris × P. acutifolius

HortScience ◽  
1990 ◽  
Vol 25 (10) ◽  
pp. 1288-1291 ◽  
Author(s):  
Ana Maria Sabja ◽  
David W.S. Mok ◽  
Machteld C. Mok
Keyword(s):  
Interspecific Hybrid ◽  
Embryo Rescue ◽  
Precocious Germination ◽  
Developmental Arrest ◽  
Hybrid Embryo ◽  
Murashige Skoog ◽  
Cotyledonary Stage ◽  
Pod Culture ◽  
Culture Procedure

Interspecific hybrid embryos resulting from crosses between Phaseolus species generally fail to reach maturity, and embryo rescue techniques are required to recover plants. To determine if ovary (pod) culture could permit interspecific hybrid embryo development, pods of P. vulgaris L. × P. acutifolius A. Gray and P. vulgaris L. (control) were cultured upright, supported by glasswool, in modified Murashige-Skoog liquid medium. The weight of seeds and length of embryos in P. vulgaris pods increased significantly during culture. However, usually only one or occasionally two seeds located at the middle positions of the pods developed to maturity. The same pod culture procedure promoted precocious germination of early cotyledonary-stage P. vulgaris × P. acutifolius embryos without further maturation of the embryos. These findings confirm that developmental arrest of the interspecific hybrid embryos in vivo is due to intrinsic deficiencies.

Interspecific hybridization of Medicago sativa L. and M. rupestris M. B. using ovule–embryo culture

1985 ◽  
Vol 27 (2) ◽  
pp. 238-245 ◽  
Author(s):  
T. J. McCoy
Keyword(s):  
Medicago Sativa ◽  
Embryo Culture ◽  
Interspecific Hybrid ◽  
Interspecific Hybrids ◽  
Embryo Rescue ◽  
Culture Method ◽  
Hybrid Plants ◽  
Successful Recovery ◽  
Hybrid Embryo ◽  
Medicago Sativa L

An ovule–embryo culture method was used to produce the first interspecific hybrids between alfalfa (Medicago sativa L.) and Medicago rupestris M. B. Culture of fertilized ovules from the cross diploid (2n = 2x = 16) M. sativa (jpjp) × diploid (2n = 2x = 16) M. rupestris began 14 days after pollination. After 5 days in culture, the interspecific hybrid embryo was removed and transferred to fresh medium, where development into a plant occurred. Forty-six M. sativa – M. rupestris F1 hybrids have been recovered using this technique. All but one of the 46 F1 hybrids were diploid (2n = 2x = 16); the only exception was tetraploid (2n = 4x = 32). The most frequent meiotic configurations observed in the F1 hybrid plants were eight bivalents or seven bivalents and two univalents, indicating significant homology between M. sativa and M. rupestris genomes. However, pollen stainability (0–12%) and pollen germination (0–6%) were extremely low. Similar to the production of the F1, no first backcross (BC1) plants were obtained from seed; however, the ovule–embryo culture method was found to be a very effective method for recovering BC1 plants and hundreds of BC1 plants have been produced. The BC1 plants from crossing the F1 with diploid M. sativa were predominantly diploid. Medicago rupestris can now be considered a potential germplasm source for alfalfa improvement. The ovule–embryo culture method represents the first successful recovery of Medicago interspecific hybrids via some form of embryo rescue. Importantly, it appears this technique can be applied to other interspecific hybrid combinations in the Medicago genus.Key words: Medicago, alfalfa, embryo culture, interspecific hybrid.

scholarly journals In vivo culture of bovine embryos and quality assessment of in vivo vs. in vitro produced embryos

2012 ◽  
Vol 50 (No. 4) ◽  
pp. 149-158 ◽  
Author(s):  
V. Havlicek ◽  
M. Lopatarova ◽  
S. Cech ◽  
R. Dolezel ◽  
T. Huber ◽  
...  
Keyword(s):  
Quality Assessment ◽  
Recovery Rate ◽  
Bovine Embryos ◽  
In Vivo Culture ◽  
Blastocyst Rate ◽  
Culture Procedure ◽  
Recovery Methods

Routine access to the bovine oviduct for in vivo culture accomplishes various demands on embryo production for scientific as well as commercial purposes. The experiments conducted in the present study focused on the efficiency of recovery methods after temporary in vivo culture of bovine embryos in oviducts of the homologous species using transvaginal endoscopy (Experiment I) and on the quality assessment of recovered blastocysts (Experiment II). In Experiment I in vitro matured oocytes were fertilized, cultured for 1 to 3 days and transferred unilaterally into the ipsilateral oviducts of 54 heifers by the means of transvaginal endoscopy. After 4 to 6 days of in vivo culture embryos were re-collected either by non-surgical flushing of uterine horns (U-group) or by combined flushing of the oviducts and uterine horns (OU-group). In total the recovery rate was 38.4% (780/2029). After flushing at day seven, 106 blastocysts (blastocyst rate: 13.6% ) were found. The additional 24 h of in vitro culture (day eight) resulted in 153 blastocysts (blastocyst rate: 19.6% ). The recovery rate in the OU-group was twice as efficient as in the U-group (390/1358 vs. 390/671, P < 0.01). The recovery rates among the different stages of transferred embryos did not differ significantly; likewise cross-effects among the stages and the recovery methods were non-significant. The recovery methods (P < 0.001) and the interaction between the recovery methods and the stages of transferred embryos (P < 0.01) had an influence on blastocyst yields on day seven (U-group 37/1358 vs. OU-group 69/671) and day eight (U-group 48/1358 vs. OU-group 105/671). In Experiment II embryo quality was assessed by the survival rate of blastocysts after freezing in ethylene glycol. Day seven embryos were produced in vitro (in vitro group D7) or by IVM/IVF followed by a combined culture procedure (2 to 3 days in vitro prior to 4 to 5 days in vivo) (in vivo group D7) or after superovulation and collection at day seven (superovulation group). Embryos from in vitro group D7 re-expanded only for 6 h after thawing, embryos from in vivo group D7 and superovulation group were alive for 24 h and 72 h of culture, respectively. Only embryos derived by superovulation showed hatching activity. Blastocysts from the in vitro group D7 and the in vivo group D7 that were held in culture medium for additional 24 h (day eight) showed an analogous post-thawing culture behaviour. In conclusion, the results of the present study demonstrated that some embryos transferred for in vivo culture remain in the oviduct even at day seven. Hence, combined flushing of oviducts and uterine horns after in vivo culture in the bovine oviduct is necessary for effective embryo re-collection. The quality of recovered embryos after temporary in vivo culture assessed by cryotolerance was in-between those produced in vitro or recovered after superovulation.

In vitro efficiency of embryo rescue of intra- and interspecific hybrid crosses of sweet cherry and Chinese cherry cultivars

2021 ◽  
Vol 275 ◽  
pp. 109716
Author(s):  
Yan-Jun Wu ◽  
Quan-Qing Song ◽  
Yue Yuan ◽  
Fang-Qi Guo ◽  
Kai-Xiang Wu ◽  
...  
Keyword(s):  
Interspecific Hybrid ◽  
Sweet Cherry ◽  
Embryo Rescue

scholarly journals Cloning, characterization and analysis by RNA interference of various genes of the Chelonus inanitus polydnavirus

2005 ◽  
Vol 86 (4) ◽  
pp. 973-983 ◽  
Author(s):  
Marianne Bonvin ◽  
Dorothee Marti ◽  
Stefan Wyder ◽  
Dejan Kojic ◽  
Marc Annaheim ◽  
...  
Keyword(s):  
Rna Interference ◽  
Host Feeding ◽  
Symbiotic Association ◽  
High Similarity ◽  
Prepupal Stage ◽  
Developmental Arrest ◽  
Larval Parasitoid ◽  
Viral Genes ◽  
Segmented Genome

Successful parasitism of some endoparasitic wasps depends on an obligately symbiotic association with polydnaviruses. These unique viruses have a segmented genome consisting of circles of double-stranded (ds) DNA and do not replicate in the parasitized host. They are produced in the wasp's ovary and injected into the host along with the egg. Chelonus inanitus is an egg–larval parasitoid; its polydnavirus (CiV) has been shown to protect the parasitoid larva from the host's immune system and to induce developmental arrest in the prepupal stage. The genome of CiV consists of at least 10–12 segments and five have been sequenced up to now. Here, the complete (CiV12g2) or partial (CiV12g1, CiV16.8g1) cloning of three new CiV genes is reported. All three occur only on one viral segment and have no similarity to other known polydnavirus genes, with the exception of a high similarity of CiV12g1 to CiV14g1 and CiV12g2 to CiV14g2. Furthermore, the first attempt of in vivo application of RNA interference to study the function of polydnavirus genes is shown. Injection of dsRNA of two late- and one early- and late-expressed CiV genes into CiV/venom-containing host eggs partially rescued last-instar larvae from developmental arrest. Injection of the same dsRNAs into parasitized eggs partially reduced parasitoid survival, mainly by preventing the successful emergence of the parasitoid from the host. These viral genes thus seem to be involved in inducing developmental arrest and in keeping the cuticle soft, which appears to be necessary for parasitoid emergence and host feeding.

In Vitro Culture of Arachis hypogaea Peg Tips1

1992 ◽  
Vol 19 (2) ◽  
pp. 78-82 ◽  
Author(s):  
Tallury P. S. Rau ◽  
H. T. Stalker ◽  
H. E. Pattee ◽  
P. Reece
Keyword(s):  
Arachis Hypogaea ◽  
Interspecific Hybrid ◽  
Embryo Rescue ◽  
Basal Medium ◽  
Early Growth ◽  
Growth Stages ◽  
Globular Stage ◽  
Arachis Hypogaea L ◽  
Indole 3 Acetic Acid

Abstract Arachis hypogaea L. cv. NC 4 was used as a model plant system in an effort to develop an in vitro embryo rescue protocol which could have application to interspecific hybrid embryos, which often abort at very early growth stages. Embryo growth and development was studied in 1- to 4-day-old peg tips containing proembryos equivalent to a stage where many interspecific hybrid embryos abort. Three independent experiments were conducted to 1) determine the most favorable basal media, 2) evaluate the effects of auxins and cytokinins on growth, and 3) determine a favorable combination of auxins and cytokinins for in vitro peanut embryo growth. The results indicated that MS (Murashige and Skoog) medium with 3% sucrose was the most favorable basal medium among seven media and two sucrose concentrations analyzed. IAA (indole-3-acetic acid) at 1.5 mg L-1 in combination with a range of KN (kinetin) levels from 0.5 to 1.25 mg L-1 were the growth regulator combinations of choice. Proembryo growth reached the multicellular globular stage, but differentiation into heart-shaped embryos did not occur.

Embryo Rescue Culture Medium for Improved Synthesis of 4x Wheat × Rye Hybrids

1994 ◽  
Vol 42 (4) ◽  
pp. 493
Author(s):  
CH Balatero ◽  
NL Darvey
Keyword(s):  
Embryo Rescue ◽  
Favourable Effect ◽  
Embryo Formation ◽  
Step Procedure ◽  
Hybrid Embryo ◽  
One Step ◽  
Well Differentiated ◽  
Semi Solid ◽  
Optimum Recovery

The cross-incompatibility barrier between 4x wheat and rye has limited the genetic base for triticale breeding. Experiments designed to improve the synthesis of wheat-rye amphihaploids were conducted. The effects of 2,4-D on crossability and 3x hybrid embryo differentiation, and the influence of one-step and two-step media on the culture of immature 3x embryos in vitro, were investigated. Application of 10 mg L-1 2,4-D slightly improved seed set but significantly reduced the frequency of normal embryos. In contrast to the reported favourable effect of 2,4-D on haploid embryo formation in wheat × maize crosses, the application of 2,4-D in the present study offers no real advantage on amphihaploid embryo formation from 4x wheat × rye crosses. For small and immature wheat-rye hybrid (3x) embryos, optimum recovery in vitro was obtained via a two-step procedure consisting of a semi-solid MN medium followed by MS medium supplemented with IAA (1 mg L-1) and BAP (1 mg L-1). For bigger and well-differentiated embryos, the use of a one-step Gamborg's B5 medium was sufficient.

scholarly journals Albinism in F1 Hybrids Hinders Geneflow Between Cultivated Chickpea (Cicer arietinum L.) and the Tertiary Gene Pool Species, Cicer Pinnatifidum

2021 ◽  
Author(s):  
Shivali Sharma ◽  
Shivaji Ajinath Lavale ◽  
Benjamin Kilian
Keyword(s):  
Gene Pool ◽  
Genetic Factors ◽  
Embryo Rescue ◽  
F1 Hybrids ◽  
Interspecific Crosses ◽  
Seed Formation ◽  
Albino Phenotype ◽  
Tertiary Gene Pool ◽  
Chickpea Cultivars

Abstract Wild Cicer species, especially those in the tertiary gene pool, carry useful alleles for chickpea improvement. The aim of this study was to evaluate the crossability and geneflow between three chickpea cultivars (as female parents) and four cross-incompatible Cicer pinnatifidum accessions (as pollen parents) from the tertiary gene pool. Ten crosses were conducted. One fully developed healthy F1 seed was harvested in vivo from the ICC 4958 × ICC 17269 cross, but the seedling developed an albino phenotype at 4–5 days after germination. Unlike other crosses, those involving the cultivar ICCV 96030 generated a large number of pods with comparatively large ovules. One albino plantlet was obtained from the ICCV 96030 × ICC 17269 cross by embryo rescue. Crosses involving ICCV 10 resulted in flower drop and poor pod set. These variable genotype-specific responses of pod, ovule, and seed development indicate that genetic factors affect the formation of interspecific hybrids. Although pod and seed formation in these interspecific crosses can be improved, geneflow between these materials is hindered by a strong genetic factor conferring albinism in the F1 hybrids.

scholarly journals Regulation of Anti–double-stranded DNA B Cells in Nonautoimmune Mice: Localization to the T–B Interface of the Splenic Follicle

1997 ◽  
Vol 186 (8) ◽  
pp. 1257-1267 ◽  
Author(s):  
Laura Mandik-Nayak ◽  
Anh Bui ◽  
Hooman Noorchashm ◽  
Ashlyn Eaton ◽  
Jan Erikson
Keyword(s):  
B Cells ◽  
Lupus Erythematosus ◽  
Turnover Rate ◽  
Cell Repertoire ◽  
Developmental Arrest ◽  
Double Stranded Dna ◽  
B Cell Repertoire ◽  
Dna Antibodies ◽  
Unique Surface

Systemic lupus erythematosus (SLE) and the MRL-lpr/lpr murine model for SLE are characterized by the presence of serum anti–double-stranded (ds)DNA antibodies (Abs), whereas nonautoimmune individuals have negligible levels of these Abs. To increase the frequency of anti-DNA B cells and identify the mechanisms involved in their regulation in nonautoimmune mice, we have used Ig transgenes (tgs). In the present study, we used the VH3H9 heavy (H) chain tg which expresses an H chain that was repeatedly isolated from anti-dsDNA Abs from MRL-lpr/lpr mice. Because the VH3H9 H chain can pair with endogenous L chains to generate anti–single-stranded DNA, anti-dsDNA, and non-DNA B cells, this allowed us to study the regulation of anti-dsDNA B cells in the context of a diverse B cell repertoire. We have identified anti-dsDNA B cells that are located at the T–B interface in the splenic follicle where they have an increased in vivo turnover rate. These anti-dsDNA B cells exhibit a unique surface phenotype suggesting developmental arrest due to antigen exposure.

Hybrid Embryo Rescue in Crop Improvement

2015 ◽  
pp. 363-384 ◽  
Author(s):  
Leela Sahijram ◽  
B. Madhusudhana Rao
Keyword(s):  
Embryo Rescue ◽  
Crop Improvement ◽  
Hybrid Embryo

Abscisic Acid and Sucrose Control of Velvetleaf (Abutilon theophrasti) Ovule Development in Vitro

Weed Science ◽  
1984 ◽  
Vol 32 (6) ◽  
pp. 798-801 ◽  
Author(s):  
Laura K. Thompson ◽  
Gerald R. Leather ◽  
Maynard G. Hale
Keyword(s):  
Abscisic Acid ◽  
Embryo Development ◽  
Parental Control ◽  
Abutilon Theophrasti ◽  
Ovule Development ◽  
Precocious Germination ◽  
Development In Vitro ◽  
Days After Anthesis

The culture of ovules excised from velvetleaf (Abutilon theophrasti Medic., ♯4 ABUTH) capsules 5 days after anthesis was used to measure the effects of abscisic acid (ABA) and sucrose on embryo development and prevention of precocious germination. ABA at 1 × 10-7 M combined with 6% sucrose in the medium for the first 14 days of culture increased embryo development but prevented precocious germination. Higher concentrations of ABA inhibited embryo development. Without ABA, precocious germination increased directly with the concentration of sucrose in the medium, and embryos died. In vivo, ABA reached its highest concentration in ovules 5 days after anthesis but was undetectable after 16 days. Parental control of embryo development may involve ABA and an increasing concentration of osmoticum as seeds dehydrate during maturation.

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