scholarly journals In vitro embryo culture to shorten the breeding cycle in lentil (Lens culinaris Medik)

2016 ◽  
Vol 127 (3) ◽  
pp. 585-590 ◽  
Author(s):  
Carolina Bermejo ◽  
Ileana Gatti ◽  
Enrique Cointry
Keyword(s):  
Embryo Culture ◽  
Lens Culinaris ◽  
Breeding Cycle

scholarly journals APPLICATION OF DIFFERENT METHODOLOGIES IN LENTIL (Lens culinaris MEDIK) BREEDING

2021 ◽  
Vol 32 (Issue 2) ◽  
pp. 32-39
Author(s):  
C.J. Bermejo ◽  
F. Maglia ◽  
T. Palacios ◽  
M.A. Espósito ◽  
F. Cazzola ◽  
...  
Keyword(s):  
Embryo Culture ◽  
Lens Culinaris ◽  
Agricultural Research ◽  
Yield Potential ◽  
Mass Selection ◽  
Breeding Methods ◽  
Genetic Base ◽  
Digital Phenotyping ◽  
New Varieties

Lentil (Lens culinaris Medik.) is a self-pollinating diploid (2n=2x=14) species belonging to the Fabaceae family. It is one of the oldest crops known, with 8,000 to 9,000 years of history and it is among the earliest domesticates from the Near East Fertile Crescent. The seeds have high nutritional value. This crop is an interesting substitute to wheat in cereal rotations but its importance is low due to a lack of suitable varieties with local adaptation. Some of the major problems that Argentinian lentil breeders face are the narrow genetic base of the current cultivated germplasm and its low yield potential. A lentil breeding program was initiated in 2004 to develop new varieties with adaptation to prevalent conditions in growing areas of Argentina. Germplasm was obtained from ICARDA (International Center for Agricultural Research in the Dry Areas) and local producers. Conventional breeding methods using hybridization and selection are being carried out to develop improved varieties, broad the genetic base, and isolate superior recombinant inbred lines. Two new varieties have been obtained, one of the macrosperm type (Boyerito FCA) and the other of the microsperm type (Tacuarita FCA) through the application of mass selection in F2 populations from the cross of selected materials. This program complements traditional breeding methods with biotechnological techniques such as transgenesis, use of molecular markers, in vitro embryo culture combined with the SSD method to shorten the breeding time, and digital phenotyping. Key words: Lentil, conventional methodologies, in vitro embryo culture, biotechnology techniques, digital phenotyping.

Multiple shoot induction in zygotic embryos: A strategy for acceleration of banana breeding

2021 ◽  
Author(s):  
Backiyarani Suthanthiram ◽  
Uma Subbaraya ◽  
Saranya Swaminathan ◽  
Durai Palani ◽  
Eugin Perianayagaraj Selvaraj ◽  
...  
Keyword(s):  
Embryo Culture ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Culture Technique ◽  
Breeding Cycle ◽  
Zygotic Embryos ◽  
Multiple Traits ◽  
Multiple Shoot Induction ◽  
Multiple Shoot Formation

Abstract The presence of residual female fertility in most of the parthenocarpic banana accessions, embryo culture technique encourages the banana breeder to develop new hybrids through conventional breeding. Desirable trait can be fixed in banana in the first generation of hybrid progenies, but their evaluation is a long process owing to non-availability of uniform suckers/planting material. The duration of the breeding cycle can be reduced by developing more number of plantlets per hybrid event under in vitro embryo culture. Thus efforts were taken to standardize the multiple shoot formation from single embryo through media manipulation and decortications of embryo culture derived single plants. Success on multiple shoot formation was achieved up to 75% and 95% in seeded accessions of M.acuminata and M.velutina embryos respectively. In general, enhanced regeneration efficiency and rate of multiple shoot formation were recorded in media with BAP concentration of 17.76µm. Single in vitro plantlets derived through embryo culture were decorticated to 3 cubic cm size that resulted in 100% multiple shoot formations. This technique has been implemented from the seeds obtained from controlled pollination (18 cross combinations) and open pollination (nine accessions) of various genomic groups (ABB, AAB, AA) and their genetic fidelity of multiple shoots were also confirmed through microsatellite markers. As this technique, i) enhances the regeneration of hybrid seeds; ii) overcomes the risk of field mortality of the hybrid progenies and iii) allows simultaneous evaluation for multiple traits, will accelerate the breeding program by reducing the time taken for the release of potential banana hybrids.

scholarly journals Modulation of Oxidative Stress and Hemostasis by Flavonoids from Lentil Aerial Parts

Molecules ◽  
2021 ◽  
Vol 26 (2) ◽  
pp. 497
Author(s):  
Jerzy Żuchowski ◽  
Agata Rolnik ◽  
Weronika Adach ◽  
Anna Stochmal ◽  
Beata Olas
Keyword(s):  
Oxidative Stress ◽  
Antioxidant Activity ◽  
Human Plasma ◽  
Crude Extract ◽  
Lens Culinaris ◽  
Bioactive Substances ◽  
Phenolic Fraction ◽  
Aerial Parts ◽  
Scientific Attention

While specific metabolites of lentil (Lens culinaris L.) seeds and their biological activity have been well described, other organs of this plant have attracted little scientific attention. In recent years, green parts of lentils have been shown to contain diverse acylated flavonoids. This work presents the results of the research on the effect of the crude extract, the phenolic fraction, and seven flavonoids obtained from aerial parts of lentils on oxidative damage induced by H2O2/Fe to lipid and protein constituents of human plasma. Another goal was to determine their effect on hemostasis parameters of human plasma in vitro. Most of the purified lentil flavonoids had antioxidant and anticoagulant properties. The crude extract and the phenolic fraction of lentil aerial parts showed antioxidant activity, only at the highest tested concentration (50 μg/mL). Our results indicate that aerial parts of lentils may be recommended as a source of bioactive substances.

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 (>84%) of concordance rate for ploidy and sex, sensitivity (>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 (>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 Znanstvena istraživanja u 21. stoljeću - etički aspekti uporabe laboratorijskih životinja i alternativni modeli

2022 ◽  
Vol 53 (5) ◽  
Author(s):  
Ivana Kmetič ◽  
Monika Roller ◽  
Marina Miletić ◽  
Teuta Murati
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Embryo Culture ◽  
Embryonic Stem ◽  
Whole Embryo Culture ◽  
Cell Test

U toksikološkim istraživanjima uz uporabu klasičnih (in vivo) istraživanja, primjenjuju se alternativni test sustavi. Korištenje laboratorijskih životinja, embrija, humanog i animalnog tkiva, kultura stanica i fetalnog seruma u istraživanjima smatra se etički problematičnim te se ograničava zakonima, pravilnicima i praksom. Razmatranjem načina kojima bi se neetičnost mogla izbjeći, došlo je do razvoja “3R” načela (akronim za tri pristupa koja bi se trebala provoditi pri istraživanjima na laboratorijskim životinjama), a to su: smanjenje/racionalizacija uporabe laboratorijskih životinja (engl. Reduction), načelo njihove zamjene (engl. Replacement) i poboljšanje uvjeta uzgoja, smještaja i skrbi za životinje (engl. Refinement). Većina je alternativnih testova toksičnosti još uvijek u postupku validacije. Pojedini in vitro testovi za istraživanja embriotoksičnosti (etički posebno osjetljivo područje) koja su priznala nadležna regulatorna tijela, su EST (engl. Embryonic Stem cell Test), WEC (engl. Whole- Embryo Culture) i MM (engl. MicroMass) test. Standardizacija protokola i uvođenje novih in vitro modela predstavlja važan segment napretka u toksikološkim istraživanjima. Znanstvena budućnost tu vidi mogućnost razvoja i implementacije načela etičnosti u istraživanja primjenjujući sustave koji će promišljeno i bez korištenja živih organizama dijelom nadomjestiti metode u biomedicini, veterinarskoj medicini, biotehnologiji i užem smislu - toksikologiji i farmakologiji.

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