scholarly journals Double Haploid Development and Assessment of Androgenic Competence of Balkan Pepper Core Collection in Bulgaria

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2414
Author(s):  
Stanislava Grozeva ◽  
Gancho Pasev ◽  
Vesela Radeva-Ivanova ◽  
Velichka Todorova ◽  
Valentina Ivanova ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Double Haploid ◽  
Genetic Improvement ◽  
Core Collection ◽  
Culture Medium ◽  
Doubled Haploids ◽  
Caps Marker ◽  
Regenerated Plants ◽  
Direct Embryogenesis ◽  
Pepper Breeding

This study was designed to assess the androgenic potential of 180 pepper accessions and 11 progenies (four F1 and seven BC) possessing PMMoV resistance in order to complement an ongoing pepper breeding program. The experiment was carried out in 10 replications with 20 anthers for each accession in two different induction mediums from 2017 to 2019. The highest androgenic response was observed in culture medium 17-2 but differences between two mediums were nonsignificant. From a total of 191 genotypes, 102 genotypes expressed a potential for direct embryogenesis. Embryo induction was seen to be genotype-dependent and decreased in the following order: Pumpkin > Conical > Bell or blocky > Round > Elongate as the most responsive genotypes with over 10% reacted anthers being observed in CAPS-23, CAPS-29, CAPS-127, CAPS-157, CAPS-169, F1 and BC 887 derived from CAPS-23. The number of regenerated plants was higher in the conical group and least in the round varietal group. Regenerated plants were examined visually and by flow cytometry for identification of spontaneous doubled haploids (DH) and haploids. Those originating from F1 and BC progenies were additionally evaluated by a CAPS marker targeting L4 allele for resistance against PMMoV. Obtained results revealed two groups consisting of homozygous susceptible and resistant plants. Therefore, use of anther culture in ongoing breeding will greatly facilitate the pepper genetic improvement.

Regeneration of double haploid plants from unpollinated ovary cultures of watermelon

2019 ◽  
Author(s):  
Yingchun Zhu ◽  
Dexi Sun ◽  
Yun Deng ◽  
Weihua Li ◽  
Guolin An ◽  
...  
Keyword(s):  
Double Haploid ◽  
Culture Medium ◽  
Chromosome Doubling ◽  
Citrullus Lanatus ◽  
Breeding Cycle ◽  
Regenerated Plants ◽  
Dark Culture ◽  
Homozygous Diploid ◽  
Optimized Conditions ◽  
Haploid Plants

Abstract Background: Watermelon (Citrullus lanatus), a major fresh fruit, is planted worldwide. Because double haploid plants may be used as parents to shorten watermelon breeding cycle, the present study optimized conditions for regenerating haploid plants from ovaries without pollination.Results: The results revealed that, the 10% sodium hypochlorite sterilized for 10 min is best for ovary enlargement. In addition, a dark culture period of 14 days promoted the ovary enlargement. The MS medium containing 0.5 mg/L NAA, 1.0 mg/L 6-BA and 0.5mg/L KT promoted the embryoid differentiation. The M2 medium containing 0.02 mg/L TDZ, 0.5 mg/L NAA, 0.5 mg/L 6-BA could be used for producing complete plants. The different genotypes affected the embryoid induction. The present study obtained regenerated plants that were established in field. Flow cytometry analyses revealed that the regenerated plants were haploid, diploid or tetraploid plant. The seedlings which were treated with culture medium can increase the chance of chromosome doubling. The SSR marker analyses showed that the diploid and tetraploid plants were homozygous at all six loci tested, indicating that these regenerated plants were double- or tetra-haploid plants.Conclusions: Haploid and homozygous diploid can be obtained through the culture of unpollinated ovary of watermelon, which is an effective way to innovate watermelon germplasm. The present study provides homozygous plants for future watermelon breeding.

scholarly journals Androgenic studies in the production of haploids and doubled haploids in Capsicum spp.

2020 ◽  
Vol 73 (1) ◽  
pp. 9047-9056 ◽  
Author(s):  
Manuel Alejandro Sánchez ◽  
Yacenia Morillo Coronado ◽  
Ana Cruz Morillo Coronado
Keyword(s):  
Genetic Variability ◽  
Double Haploid ◽  
Genetic Improvement ◽  
Doubled Haploids ◽  
Horticultural Crop ◽  
Breeding Programs ◽  
Agronomic Characteristics ◽  
The World ◽  
Capsicum Spp ◽  
The Tropics

Capsicum spp. is a horticultural crop of agronomic interest and is considered the fourth most important vegetable in the world. It is an important nutritional and medicinal source, and its production generates employment in the tropics. In this species, the genetic variability is wide and with great potential, which has been exploited to generate outstanding varieties. Breeding programs seek different alternatives to accelerate the production of improved varieties with desirable agronomic characteristics. These objectives can be achieved with the production of haploid and double haploid plants via androgenesis or gynogenesis, being androgenesis the approach most used for paprika cultures. The purpose of this review is to present the results of different researches in obtaining haploids and doubled haploids in cultivars of Capsicum spp. and its impact on the genetic improvement of this crop. 

scholarly journals Morphogenesis of Anthers Isolated From Cabbage (Brassica Oleracea L.) In Vitro

2022 ◽  
Author(s):  
Rima Kirakosyan ◽  
Elena Kalashnikova
Keyword(s):  
Culture Medium ◽  
Root Meristem ◽  
Reproductive Organs ◽  
Biologically Active ◽  
Regenerated Plants ◽  
Direct Embryogenesis ◽  
Morphogenetic Potential ◽  
Cytological Method ◽  
Active Substances

This study aimed to optimize the steps of obtaining regenerated cabbage plants by direct embryogenesis from isolated anthers and ovaries. Stepwise pretreatment of inflorescences was usedfor the studied hybrids and inbred lines. First, the inflorescences were placed in water and kept at a temperature of +4-6∘C for one day without the use of biologically active substances. Then the inflorescences were placed in a solution of the drug Dropp (10 mg/l) and cultivated for two days. After that, the anthers and ovaries were isolated from the flower buds and cultured on the MS culture medium at a temperature of + 32∘C for one day. The cultivation of the isolated explants on a nutrient medium (containing 0.01 mg/lof Dropp, 1.0 mg/lof NAA, 500 mg/lof asparagine, 100 mg/l of tyrosine, and 10 g/l of sucrose)led to an increase in their morphogenetic potential in the culture of anthers and ovaries (by 3.42% and 5.54%, respectively).A cytological method was usedto demonstrate the haploid nature of the regenerating plants. The number of chromosomes in the root meristem andleaves, and the chloroplasts in the closing cells of the stomatawere calculated. Keywords: cabbage, culture in vitro, regenerated plants, anthers, ovaries, reproductive organs

scholarly journals Cytokinin Mediated Increased In Vitro Production of Secondary Metabolites with Special Reference to Solasodine in Solanum erianthum

2021 ◽  
Vol 8 (02) ◽  
pp. e62-e68
Author(s):  
Jeeta Sarkar ◽  
Nirmalya Banerjee
Keyword(s):  
Secondary Metabolites ◽  
High Performance ◽  
Culture Medium ◽  
In Vitro Production ◽  
Leaf Extracts ◽  
Plant Parts ◽  
Regenerated Plants ◽  
Vitro Production ◽  
Regenerated Plantlets

AbstractSteroid alkaloid solasodine is a nitrogen analogue of diosgenin and has great importance in the production of steroidal medicines. Solanum erianthum D. Don (Solanaceae) is a good source of solasodine. The aim of this study was to evaluate the effect of different cytokinins on the production of secondary metabolites, especially solasodine in the in vitro culture of S. erianthum. For solasodine estimation, field-grown plant parts and in vitro tissues were extracted thrice and subjected to high-performance liquid Chromatography. Quantitative analysis of different secondary metabolites showed that the amount was higher in the in vitro regenerated plantlets compared to callus and field-grown plants. The present study critically evaluates the effect of the type of cytokinin used in the culture medium on solasodine accumulation in regenerated plants. The highest solasodine content (46.78±3.23 mg g-1) was recorded in leaf extracts of the in vitro grown plantlets in the presence of 6-γ,γ-dimethylallylamino purine in the culture medium and the content was 3.8-fold higher compared to the mother plant.

scholarly journals ASSESSMENT OF GENETIC STABILITY OF MICROPROPAGATED Eucalyptus globulus Labill HYBRID CLONES BY MEANS OF FLOW CYTOMETRY AND MICROSATELLITES MARKERS

2017 ◽  
Vol 41 (1) ◽  
Author(s):  
Leandro Silva Oliveira ◽  
Aloisio Xavier ◽  
Wagner Campos Otoni ◽  
José Marcello Salabert Campos ◽  
Lyderson Facio Viccini ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Microsatellite Markers ◽  
Genetic Stability ◽  
Culture Medium ◽  
Eucalyptus Grandis ◽  
Genetic Fidelity ◽  
Shoot Multiplication ◽  
Micropropagated Plants ◽  
Hybrid Clones

ABSTRACT Flow cytometry and microsatellite markers were used to determine a genetic fidelity of micropropagated plants from the two Eucalyptus urophylla x E. globulus clones and a Eucalyptus grandis x E. globulus clone derived from adult material. Clones were repeatedly subcultured for 25 subcultures on MS medium supplemented with BA (2.22 µM) and ANA (0.05 µM) for in vitro shoot multiplication. The elongation was performed in MS culture medium supplemented with AIB (2.46 µM) and BA(0.22 µM). The ex vitro rooting and acclimatization phases were lead at the same time. The micropropagated clones showed genetic stability by flow cytometry and microsatellite markers. The results proved that micropropagation, for purposes of rejuvenation, can be a viable technique to generate genetically stable or identical E. globulus hybrid clones.

scholarly journals Improved the Agrobacterium tumefaciens-mediated transformation of cucumber by a modified the using of antibiotics and acetosyringone

2020 ◽  
Author(s):  
Li'ang Chai ◽  
Changxia Du ◽  
Huaifu Fan ◽  
Chen Liu ◽  
Yuyang Si
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Culture Medium ◽  
Early Stage ◽  
Marker Gene ◽  
Vegetable Crops ◽  
Regenerated Plants ◽  
Npt Ii ◽  
Screening Efficiency ◽  
Speed Up ◽  
Polymerase Chain

Abstract Background: Cucumber (Cucumis sativus) is one of the most important vegetable crops in the world. As conventional breeding of cucumber is very challenging, genetic engineering is an alternative option to introduce important traits such as enhanced stress resistance and nutritional value. However, the efficiency of the transformation system depends on genotypes, transformation conditions, selection agents, etc. This study aims to speed up the process of Agrobacterium-mediated transformation of cucumber. ‘ Xintai mici ’, a very popular and typical north China-type cucumber variety, was transformed with Agrobacterium GV3101. The strain carried pCAMBIA2300s plasmid, a double vector with the marker gene of neomycin phosphotransferase II ( npt II). Results: The research results indicated that cefotaxime sodium was suitable for inhibiting Agrobacterium in the stage of screening and bud elongation. Timentin was best used during rooting stage. Furthermore, 25 mg/L kanamycin was used in the early stage of screening and increased to 50 mg/L for further screening. At the bud elongation and rooting stage, 75 and 100 mg/L kanamycin was used respectively to improve the screening efficiency. In order to obtain the highest regeneration frequency of resistant buds, 50, 150, and 100 μM acetosyringone were added in the pre-culture medium, infection solution, and co-culture medium respectively. To confirm the presence of the transgenes, DNA from npt II transgenic cucumber plants was analyzed by polymerase chain reaction after transplanting resistant regenerated plants. Conclusions: We finally achieved an 8.1% conversion, which was among the highest values reported until date using cucumber ‘ Xintai mici ’. Thus an effective protocol for Agrobacterium tumefaciens -mediated genetic transformation of cucumber was optimized.

scholarly journals Comparison of Generic Fluorescent Markers for Detection of Extracellular Vesicles by Flow Cytometry

2018 ◽  
Vol 64 (4) ◽  
pp. 680-689 ◽  
Author(s):  
Leonie de Rond ◽  
Edwin van der Pol ◽  
Chi M Hau ◽  
Zoltan Varga ◽  
Auguste Sturk ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Extracellular Vesicles ◽  
Culture Medium ◽  
Breast Adenocarcinoma ◽  
Analytical Sensitivity ◽  
Adenocarcinoma Cell Line ◽  
Acetoxymethyl Ester ◽  
Fluorescent Markers ◽  
Conditioned Culture Medium ◽  
Carboxyfluorescein Succinimidyl Ester

Abstract BACKGROUND Extracellular vesicles (EVs) in biofluids are potential biomarkers of disease. To explore the clinical relevance of EVs, a specific generic EV marker would be useful, one that does not require antibodies and binds to all EVs. Here we evaluated 5 commonly used generic markers for flow cytometry. METHODS Flow cytometry (A60-Micro, Apogee) was used to evaluate the ability of the generic EV markers calcein acetoxymethyl ester, calcein acetoxymethyl ester violet, carboxyfluorescein succinimidyl ester (CFSE), 4-(2-[6-(dioctylamino)-2-naphthalenyl]ethenyl)-1-(3-sulfopropyl)pyridinium (di-8-ANEPPS), and lactadherin to stain EVs from MCF7 human breast adenocarcinoma cell line-conditioned culture medium [epithelial cell adhesion molecule positive (EpCAM+)] or platelet EVs from human plasma [integrin β3 positive (CD61+)]. Side scatter triggering was applied as a reference, and the influence of non-EV components (proteins and lipoproteins) was evaluated. RESULTS Di-8-ANEPPS, lactadherin, and side scatter detected 100% of EpCAM+ MCF7 EVs. Lactadherin and side scatter detected 33% and 61% of CD61+ EVs, respectively. Di-8-ANEPPS detected platelet EVs only if soluble protein was first removed. Because all generic markers stained proteins, at best 33% of platelet EVs in plasma were detected. The calcein markers and CFSE were either insensitive to EVs in both samples or associated with swarm detection. CONCLUSIONS None of the generic markers detected all and only EVs in plasma. Side scatter triggering detected the highest concentration of plasma EVs on our A60-Micro, followed by lactadherin. The choice between scatter or lactadherin primarily depends on the analytical sensitivity of the flow cytometer used.

scholarly journals Development of a Core Collection of Strawberry Cultivars Based on SSR and CAPS Marker Polymorphisms

2017 ◽  
Vol 86 (3) ◽  
pp. 365-378 ◽  
Author(s):  
Takuya Wada ◽  
Yuji Noguchi ◽  
Sachiko Isobe ◽  
Miyuki Kunihisa ◽  
Takayuki Sueyoshi ◽  
...  
Keyword(s):  
Core Collection ◽  
Caps Marker ◽  
Strawberry Cultivars

scholarly journals Androgenesis of Red Cabbage in Isolated Microspore Culture In Vitro

Plants ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1950
Author(s):  
Anna Mineykina ◽  
Ludmila Bondareva ◽  
Alexey Soldatenko ◽  
Elena Domblides
Keyword(s):  
Culture Medium ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Antioxidant Properties ◽  
F1 Hybrids ◽  
Anthocyanin Content ◽  
Isolated Microspore Culture ◽  
Culture In Vitro ◽  
Red Cabbage

Red cabbage belongs to the economically important group of vegetable crops of the Brassicaceae family. A unique feature of this vegetable crop that distinguishes it from other members of the family is its unique biochemical composition characterized by high anthocyanin content, which gives it antioxidant properties. The production mainly uses F1 hybrids, which require constant parental lines, requiring 6–7 generations of inbreeding. Culture of isolated microspores in vitro is currently one of the promising methods for the accelerated production of pure lines with 100% homozygosity. The aim of this study is to investigate the factors and select optimal parameters for successful induction of red cabbage embryogenesis in isolated microspore culture in vitro and subsequent regeneration of DH plants. As a result of research, for the first time, it was possible to carry out the full cycle of obtaining DH plants of red cabbage from the induction of embryogenesis to their inclusion in the breeding process. The size of buds containing predominantly microspores at the late vacuolated stage and pollen at the early bi-cellular stage has to be selected individually for each genotype, because the embryoid yield will be determined by the interaction of these two factors. In the six samples studied, the maximum embryoid yield was obtained from buds 4.1–4.4 mm and 4.5–5.0 mm long, depending on the genotype. Cultivation of microspores was carried out on liquid NLN culture medium with 13% sucrose. The maximum number of embryoids (173.5 ± 7.5 pcs./Petri dish) was obtained on culture medium with pH 5.8 and heat shock at 32 °C for 48 h. Successful embryoid development and plant regeneration by direct germination from shoot apical meristem were achieved on MS culture medium with 2% sucrose and 0.7% agar, supplemented with 6-benzylaminopurine at a concentration of 1 mg/L. Analysis of the obtained regenerated plants, which successfully passed the stage of adaptation to ex vitro conditions by flow cytometry, showed that most of them were doubled haploids (up to 90.9%). A low number of seeds produced by self-fertilization in DH plants was observed.

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