Muskmelon Embryo Rescue Techniques Using In Vitro Embryo Culture

2010 ◽  
pp. 107-115
Author(s):  
Hector Gordon Nuñez-Palenius ◽  
Rafael Ramírez-Malagón ◽  
Neftalí Ochoa-Alejo
Keyword(s):  
Embryo Culture ◽  
Embryo Rescue

scholarly journals Reproductive Behavior of Induced Allotetraploid ×Chitalpa and In Vitro Embryo Culture of Polyploid Progeny

2006 ◽  
Vol 131 (6) ◽  
pp. 716-724 ◽  
Author(s):  
Richard T. Olsen ◽  
Thomas G. Ranney ◽  
Zenaida Viloria
Keyword(s):  
Embryo Culture ◽  
Embryo Rescue ◽  
Intergeneric Hybrid ◽  
Culture Methods ◽  
Ploidy Levels ◽  
Tetraploid Level ◽  
Chilopsis Linearis ◽  
Basal Salts ◽  
Excised Embryos

×Chitalpa tashkentensis Elias & Wisura is a sterile intergeneric hybrid [Catalpa bignonioides Walt. × Chilopsis linearis (Cav.) Sweet]. To restore fertility in ×Chitalpa the following were evaluated: 1) oryzalin as a polyploidization agent, 2) fertility of induced polyploids, and 3) in vitro culture methods for embryo rescue of interploid crosses. Meristems of ×Chitalpa `Pink Dawn' were submerged in an aqueous solution of 150 μm oryzalin for 0, 6, 12, or 24 hours and ploidy analyzed via flow cytometry. As treatment duration increased, recovery of diploids decreased as mixoploids and shoot mortality increased. Two tetraploid shoots occurred in the 24-hour treatment. Four tetraploids and two cytochimeras were stabilized in total. Tetraploids flowered sparsely; however, cytochimeras flowered profusely and these were used to study fertility at the tetraploid level. Diploid ×Chitalpa `Pink Dawn' pollen was essentially nonviable, but cytochimera pollen stained and germinated equal to or greater than pollen of C. bignonioides and C. linearis `Bubba'. Cytochimera ×Chitalpa were selfed yielding tetraploid seedlings, crossed with C. bignonioides to yield triploids, but failed in reciprocal crosses with C. linearis `Bubba' and `Burgundy Lace'. To increase recovery of triploids, germination of triploid and tetraploid embryos was investigated, as either intact ovules or excised embryos, on Schenk and Hildebrandt (SH) basal salts supplemented with sucrose at 20, 40, and 80 g·L-1, presence or absence of 2% coconut-water, and gibberellic acid (GA3) at 0, 1, 2, or 4 μm, and harvested weekly beginning 2 weeks after pollination (WAP). Germination of triploids (cytochimera ×Chitalpa × diploid C. bignonioides) and tetraploids (selfed cytochimera ×Chitalpa) were greatest with excised embryos at 7 WAP on SH supplemented with sucrose at 20 g·L-1 and ≥1 μm GA3. Germination of triploids (diploid C. linearis × cytochimera ×Chitalpa) was <5% at 4, 5, or 6 WAP on the same medium as above. Oryzalin effectively induced polyploidy and restored fertility in ×Chitalpa `Pink Dawn'. Successful crosses between hybrid and parental taxa of different ploidy levels, coupled with embryo culture will facilitate a ×Chitalpa breeding program. Chemical names used: 4(dipropylamino)-3,5-dinitrobenzenesulfonamide (oryzalin).

scholarly journals Embryo Culture and Embryo Rescue in Brassica

2021 ◽  
Author(s):  
Mohammad Akmal
Keyword(s):  
Embryo Culture ◽  
Interspecific Hybrids ◽  
Higher Plants ◽  
Zygotic Embryo ◽  
Embryo Rescue ◽  
Culture Conditions ◽  
Reproductive Barriers ◽  
Whole Plant ◽  
Embryo Regeneration

Somatic embryogenesis is the best demonstration of totipotency in higher plants in which somatic cell produce whole plant like zygotic embryo. It is also demonstrated that immature, weak, hybrid or sometimes inviable embryos can be saved through in vitro culture to prevents its degradation. It may help to cross the reproductive barriers when interspecific hybrids developed. Brasssica is an economically valuable oil yielding and vegetable crop and India is the largest producer of oil seed rape in the world. Various factors affect the embryo rescue in Brassica like growth stage of the embryos, types and composition of the rescue medium etc. The embryo regeneration potential can improve through the modification of culture conditions in both zygotic as well as somatic embryo. Except the embryo culture other parts like ovule, ovary culture can also be done to developed interspecific hybrids. This chapter is focused on the embryo rescue techniques in the genus Brassica and summarizes possible ways of improving the technique used.

P–582 High level of concordance between invasive and non-invasive preimplantation genetic testing for aneuploidies (niPGT-A) at day5 and day6–7

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
A Biricik ◽  
V Bianchi ◽  
F Lecciso ◽  
M Surdo ◽  
M Manno ◽  
...  
Keyword(s):  
Data Analysis ◽  
Embryo Culture ◽  
Culture Media ◽  
Embryo Biopsy ◽  
Non Invasive ◽  
Culture Time ◽  
Ngs Data Analysis ◽  
High Level ◽  
Ngs Data

Abstract Study question To explore ploidy concordance between invasive and non-invasive PGTA (niPGT-A) at different embryo culture time. Summary answer High level (&gt;84%) of concordance rate for ploidy and sex, sensitivity (&gt;88%), and specificity (76%) were obtained for both day6/7 samples and day5 samples. What is known already The analysis of embryo cell free DNA (cfDNA) that are released into culture media during in vitro embryo development has the potential to evaluate embryo ploidy status. However, obtaining sufficient quality and quantity of cfDNA is essential to achieve interpretable results for niPGT-A. More culture time is expected to be directly proportional to the release of more cfDNA. But embryo culture time is limited due to in-vitro embryo survival potential. Therefore, it is important to estimate the duration of the culture that will provide the maximum cfDNA that can be obtained without adversely affecting the development of the embryo. Study design, size, duration A total of 105 spent culture media (SCM) from day5-day7 blastocyst stage embryos have been included in this cohort study. The cfDNA of SCM samples were amplified and analyzed for niPGT-A by NGS analysis. The SCM samples were divided into 2 subgroups according the embryo culture hours (Day5 and Day6/7 group). The DNA concentration, informativity and euploidy results have then been compared with their corresponding embryos after trophectoderm biopsy (TE) and PGT-A analysis by NGS Participants/materials, setting, methods Embryos cultured until Day3 washed and cultured again in 20µl fresh culture media until embryo biopsy on Day5, 6, or 7. After biopsy SCM samples were immediately collected in PCR tubes and conserved at –20 °C until whole genome amplification by MALBAC® (Yicon Genomics). The TE and SCM samples were analyzed by next-generation sequencing (NGS) using Illumina MiSeq® System. NGS data analysis has been done by Bluefuse Multi Software 4.5 (Illumina) for SCM and TE samples Main results and the role of chance Only the SCM samples which have an embryo with a conclusive result were included in this cohort (n = 105). Overall 97.1% (102/105) of SCM samples gave a successful DNA amplification with a concentration ranging 32.4–128.5ng/µl. Non-informative (NI) results including a chaotic profile (&gt;5 chromosome aneuploidies) were observed in 17 samples, so 83.3%(85/102) of SCM samples were informative for NGS data analysis. Ploidy concordance rate with the corresponding TE biopsies (euploid vs euploid, aneuploid vs aneuploid) was 84.7% (72/85). Sensitivity and specificity were 92,8% and 76,7%, respectively with no significant difference for all parameters for day 6/7 samples compared with day 5 samples. The false-negative rate was 3.5% (3/85), and false-positive rate was 11.7% (10/85). Limitations, reasons for caution The sample size is relatively small. Larger prospective studies are needed. As this is a single-center study, the impact of the variations in embryo culture conditions can be underestimated. Maternal DNA contamination risk cannot be revealed in SCM, therefore the use of molecular markers would increase the reliability. Wider implications of the findings: Non-invasive analysis of embryo cfDNA analyzed in spent culture media demonstrates high concordance with TE biopsy results in both early and late culture time. A non-invasive approach for aneuploidy screening offers important advantages such as avoiding invasive embryo biopsy and decreased cost, potentially increasing accessibility for a wider patient population. Trial registration number Not applicable

scholarly journals In vitro embryo rescue and plant regeneration following self-pollination with irradiated pollen in cassava (Manihot esculenta Crantz)

2015 ◽  
Vol 14 (27) ◽  
pp. 2191-2201
Author(s):  
Buttibwa Mary ◽  
S Kawuki Robert ◽  
K Tugume Arthur ◽  
Akol Jacinta ◽  
Magambo Stephen ◽  
...  
Keyword(s):  
Plant Regeneration ◽  
Manihot Esculenta ◽  
Embryo Rescue ◽  
Manihot Esculenta Crantz ◽  
Irradiated Pollen ◽  
Self Pollination

scholarly journals Adaptability and leaf anatomical features in oil palm seedlings produced by embryo rescue and pre-germinated seeds

2010 ◽  
Vol 22 (3) ◽  
pp. 209-215 ◽  
Author(s):  
Zanderluce G. Luis ◽  
Kadja Milena G. Bezerra ◽  
Jonny Everson Scherwinski-Pereira
Keyword(s):  
Oil Palm ◽  
Embryo Rescue ◽  
Morphological Characteristics ◽  
Growth Conditions ◽  
Epidermal Cells ◽  
In Vitro Cultivation ◽  
Vascular Bundles ◽  
Anatomical Plasticity ◽  
Germinated Seeds

Changes in the leaf structure of plants grown in different conditions have been reported, such as increase in size and density of stomata and reduction in stomatal control, amount of epicuticular wax, and mesophyll thickness, with a high diversity of intercellular spaces. However, these changes are highly variable depending on the physiological and morphological characteristics of each species. The objective of this work was to analyze the adaptability and anatomical plasticity of oil palm seedlings produced after embryo rescue and pre-germinated seeds. Expanded leaves were prepared for evaluation of morphometric data and anatomical structures. It was verified that the environmental conditions in vitro negatively influenced the stomata density, epidermal and hypodermal thickness, and the values for the expansion cells and leaf mesophile. Anatomically, the oil palm leaves present the same tissues composition in both growth conditions, with uniseriate epidermal cells, and tetracitic stomata occurring in both epidermal surfaces. Epidermal cells from in vitro plants are thinner than ones from greenhouse. The midrib of leaves from greenhouse plants are more developed and is composed by only one central vascular bundle, while plants from in vitro cultivation developed three to four collateral vascular bundles.

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