scholarly journals Efficient Tissue Culture Protocol for Magnolia lucida (Magnoliaceae) and Confirmation of Genetic Stability of the Regenerated Plants

Plants ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 997
Author(s):  
Lu Kang ◽  
Keyuan Zheng ◽  
Yuqing Xie ◽  
Yanwen Deng ◽  
Yina Yu ◽  
...  
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Continuous Light ◽  
Inter Simple Sequence Repeat ◽  
Parent Material ◽  
Naphthaleneacetic Acid ◽  
Multiplication Rate ◽  
Regenerated Plants ◽  
Benzalkonium Bromide ◽  
Optimal Medium

Magnolia lucida (Magnoliaceae) is classified as an endangered species by the International Union for Conservation of Nature. It has high commercial value owing to its attractive tree shape and flowers. We adopted an excellent genotype of M. lucida as the parent material and established a mini-cut orchard through grafting to provide trunk shoots explants over the long-term. Optimal sterilization was achieved using a combination of 75% ethanol for 30 s, one percent benzalkonium bromide for five minutes, and 0.1% mercuric chloride for five minutes. Modified Murashige and Skoog medium (ML) was the optimal medium for the growth of M. lucida. Addition of one mg/L of 6-benzyl adenine (BA) and 0.05 mg/L of α-naphthaleneacetic acid (NAA) to the medium increased the shoot induction rate to 95.56%, and the ML medium containing 0.4 mg/L BA and 0.04 mg/L NAA achieved the maximum multiplication rate (284.56%). Dark treatment for seven days, followed by continuous light treatment could better resolve the challenge of difficult rooting in M. lucida plants. Using random amplified polymorphic DNA and inter simple sequence repeat markers, we confirmed the genetic uniformity and stability of the regenerated plants. Our protocol should be helpful for the propagation and conservation of this endangered plant.

scholarly journals Micropropagation of Inula germanica L. from the Seedlings Explants

2018 ◽  
Vol 46 (1) ◽  
pp. 52-57 ◽  
Author(s):  
Alina TREJGELL ◽  
Monika KAMIŃSKA ◽  
Karolina LISOWSKA ◽  
Andrzej TRETYN
Keyword(s):  
Solid Medium ◽  
Positive Impact ◽  
Shoot Tips ◽  
Fluorescent Light ◽  
First Year ◽  
Naphthaleneacetic Acid ◽  
Multiplication Rate ◽  
Indolebutyric Acid ◽  
Regenerated Plants

This is the first communication of micropropagation system for Inula germanica using seedling explants germinated in vitro. The development of this system gives the possibility of future reintroduction of I. germanica providing a way to stabilize or re-establish its population. Shoot tips and fragments of cotyledons, hypocotyls and roots were isolated from ten-day-old seedlings. Explants were put on MS medium containing 1.0 mg l-1 benzylaminopurine and 0.1 mg l-1 naphthaleneacetic acid and cultured under continuous white fluorescent light (45 μmol.m-2.s-1) at 26 ± 1 °C. The highest percentage of shoot organogenesis (83.3%) was recorded for hypocotyl, while the highest average number of shoots per explant (12.0) was recorded for shoot tips. In subsequent subcultures, multiplication rate decreased to 3.0-4.9 shoots per explant. Less than 19% shoots were able to root on the solid medium without auxins. The highest rooting efficiency (69.3%) was recorded for solid medium supplemented with indolebutyric acid, but growth of roots was inhibited. The percentage of rooted shoots (62.2%) and number of roots per shoot (2.4 per shoot) into the liquid medium were comparable to medium with 0.1 mg·l-1 indolebutyric acid. showing a positive impact on the process of acclimatization. The regenerated plants were able to flowering in the first year after acclimatization. Developed micropropagation system for I. germanica is efficient and can be a useful tool for the active protection of this species.

Micropropagation and assessment of genetic stability in Celastrus paniculatus: An endangered medicinal plant

Biologia ◽  
2013 ◽  
Vol 68 (4) ◽  
Author(s):  
Sunil Senapati ◽  
Subhashree Aparajita ◽  
Gyana Rout
Keyword(s):  
Medicinal Plant ◽  
Genetic Stability ◽  
In Vitro Regeneration ◽  
Random Amplified Polymorphic Dna ◽  
Basal Medium ◽  
Naphthaleneacetic Acid ◽  
Multiplication Rate ◽  
Endangered Medicinal Plant ◽  
Celastrus Paniculatus

AbstractA highly efficient protocol for in vitro regeneration of an indigenous, endangered medicinal plant Celastrus paniculatus was achieved using nodal explants. Murashige and Skoog (MS) basal medium supplemented with 0.5 mg/L 6-benzylaminopurine (BAP) and 0.1 mg/L naphthaleneacetic acid (NAA) showed maximum percentage of shoot multiplication (83.4%) with 8.2 shoots/explants. Maximum rooting of 73.3% with 4.8 roots/shoot was achieved on half-strength MS media supplemented with 0.5 mg/L indole-3-acetic acid (IAA) and the percentage of survival was 91% after acclimatization. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) marker study confirmed genetic stability for in vitro raised explants by showing 100% monomorphism. High multiplication rate associated with genetic stability ensure the efficacy of the present in vitro clonal propagation protocol of this important medicinal plant species.

scholarly journals Genetic Stability of the Endangered Species Salix lapponum L. Regenerated In Vitro during the Reintroduction Process

Biology ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 378
Author(s):  
Marzena Parzymies ◽  
Magdalena Pogorzelec ◽  
Katarzyna Głębocka ◽  
Elwira Śliwińska
Keyword(s):  
Genetic Stability ◽  
Inter Simple Sequence Repeat ◽  
Multiplication Rate ◽  
Natural Habitats ◽  
Relict Species ◽  
Flow Cytometric ◽  
Regenerated Plants ◽  
Simple Sequence ◽  
Salix Lapponum

Salix lapponum L. is a boreal relict species, threatened with extinction in Poland. An 80% decrease in the number of its stands was confirmed in the last half-century, so that to prevent the loss of downy willow, attempts were made to reintroduce this species in natural habitats. Micropropagation was chosen as a first stage of its active conservation. S. lapponum shoots were collected and disinfected with NaOCl, AgNO3, or HgCl2 or with a two-step disinfection with NaOCl and then placed on MS medium with BA 1 mg·dm−3 and IBA 0.1 mg·dm−3. Regenerated shoots were cultivated with addition of BA, KIN, or 2iP, alone or in combination with auxins, to find the highest multiplication rate. Inter-simple sequence repeat (ISSR) analysis and flow cytometric analyses were conducted on in vitro regenerated plants to check their genetic stability. Disinfection was quite difficult and the use of HgCl2 was the most efficient. The highest multiplication rate was obtained in presence of KIN at 0.5 mg·dm−3 + IAA at 0.5 mg·dm−3. The analysis confirmed the genome size stability, which is in agreement with the results obtained by ISSR, revealing no somaclonal variation in plantlets and therefore allowing the use of the obtained plants for reintroduction.

Long-term in vitro Storage Technique of Valuable Birch Genotypes and Plant Production on its Basis

2019 ◽  
pp. 57-67
Author(s):  
T.M. Tabatskaya ◽  
N.I. Vnukova
Keyword(s):  
Plant Production ◽  
High Survival ◽  
Ex Vitro ◽  
Regenerative Capacity ◽  
In Vitro Storage ◽  
Interspecific Differences ◽  
Regenerated Plants ◽  
Growing Conditions

A technique for the long-term (up to 27 years) in vitro storage of valuable birch genotypes under normal (25 °C, 2.0 klx, 16-h day and 8-h night) and low temperature (4 °C, 0.5 klx, 6-h day and 18-h night) growing conditions on hormone-free media has been described. The study explored for the first time the influence of different strategies to store the clones of Betula pubescens and B. pendula var. сarelica (6 genotypes) on the regenerative capacity of collection samples, adaptive potential of regenerated plants and plant production by the in vitro and ex vitro techniques. It was established that both storage strategies provided a persistently high survival rate (82-100%) and regenerative capacity of in vitro shoots (the multiplication coefficient of 4.2-6.3 and rhizogenic activity of 90-100%). The clones retained their characteristics of height growth under the in vitro and ex vitro conditions, and demonstrated intraclonal homogeneity and lack of signs of somaclonal variability. The plants showed substantial interspecific differences at the stage of multiplication and transfer to the greenhouse. The highest percentage of acclimated plants (75-98% depending on the clone genotype) was obtained after planting of micro plants straight in the greenhouse, which simplified the technology and made plant production less costly. long-term in vitro storage, birch, species, genotype, micropropagation, ex vitro adaptation, plant material

scholarly journals Long term results of accelerated 9 mW corneal crosslinking for early progressive keratoconus: the Siena Eye-Cross Study 2

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Cosimo Mazzotta ◽  
Frederik Raiskup ◽  
Farhad Hafezi ◽  
Emilio A Torres-Netto ◽  
Ashraf Armia Balamoun ◽  
...  
Keyword(s):  
Visual Acuity ◽  
Corneal Thickness ◽  
Continuous Light ◽  
Clinical Results ◽  
Asymmetry Index ◽  
Long Term Results ◽  
Distance Visual Acuity ◽  
Long Term Stability ◽  
Uncorrected Distance Visual Acuity

Abstract Purpose To assess clinical results of the 9 mW/5.4 J/cm2 accelerated crosslinking (ACXL) in the treatment of progressive keratoconus (KC) over a span of 5 years. Methods The prospective open non-randomized interventional study (Siena Eye-Cross Study 2) included 156 eyes of 112 patients with early progressive KC undergoing the Epi-Off 9 mW/5.4 J/cm2 ACXL at the Siena Crosslinking Centre, Italy. The mean age was 18.05 ± 5.6 years. The 20-min treatments were performed using the New KXL I (Avedro, Waltham, USA), 10 min of 0.1% HPMC Riboflavin soaking (VibeX Rapid, Avedro, Waltham, USA) and 10 min of continuous-light UV-A irradiation. Uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), Kmax, coma, minimum corneal thickness (MCT), surface asymmetry index (SAI), endothelial cell count (ECC) were measured, and corneal OCT performed. Results UDVA and CDVA improved significantly at the 3rd (P = 0.028), Δ + 0.17 Snellen lines and 6th postoperative month, respectively (P < 0.001), Δ + 0.23 Snellen lines. Kmax improved at the 6th postoperative month (P = 0.03), Δ − 1.49 diopters from the baseline value. Also, coma aberration value improved significantly (P = 0.004). A mild temporary haze was recorded in 14.77% of patients without affecting visual acuity and without persistent complications. Corneal OCT revealed a mean demarcation line depth at 332.6 ± 33.6 μm. Conclusion The 5-year results of Epi-Off 9 mW/5.4 J/cm2 ACXL demonstrated statistically significant improvements in UCVA and CDVA, corneal curvature and corneal higher-order aberrations which confers a long-term stability for progressive ectasia. Based on the results of the Siena Eye-Cross Study 2, the 9 mW/5.4 J/cm2 ACXL is a candidate to be  the natural evolution of Epi-Off CXL treatment for the management of early progressive corneal ectasia, and thus optimize clinic workflow.

Species-diagnostic and species-specific DNA sequences evenly distributed throughout pine and spruce chromosomes

Genome ◽  
2010 ◽  
Vol 53 (10) ◽  
pp. 769-777 ◽  
Author(s):  
Melanie Mehes-Smith ◽  
Paul Michael ◽  
Kabwe Nkongolo
Keyword(s):  
Dna Sequences ◽  
Random Amplified Polymorphic Dna ◽  
Inter Simple Sequence Repeat ◽  
Issr Markers ◽  
Pinus Monticola ◽  
The Family ◽  
Species Specific ◽  
Rapd And Issr ◽  
Specific Sequences

Genome organization in the family Pinaceae is complex and largely unknown. The main purpose of the present study was to develop and physically map species-diagnostic and species-specific molecular markers in pine and spruce. Five RAPD (random amplified polymorphic DNA) and one ISSR (inter-simple sequence repeat) species-diagnostic or species-specific markers for Picea mariana , Picea rubens , Pinus strobus , or Pinus monticola were identified, cloned, and sequenced. In situ hybridization of these sequences to spruce and pine chromosomes showed the sequences to be present in high copy number and evenly distributed throughout the genome. The analysis of centromeric and telomeric regions revealed the absence of significant clustering of species-diagnostic and species-specific sequences in all the chromosomes of the four species studied. Both RAPD and ISSR markers showed similar patterns.

scholarly journals Jacob’s ladder Polemonium caeruleum L. and black salsify Sсorzonera hispanica L. in vitro culture

2018 ◽  
Vol 22 ◽  
pp. 216-221
Author(s):  
O. V. Bulko ◽  
L. G. Lioshina
Keyword(s):  
In Vitro Culture ◽  
Root Culture ◽  
Medium Composition ◽  
Cultivation Medium ◽  
Plant Cultivation ◽  
Regenerated Plants ◽  
Composition Modification ◽  
Jacob's Ladder ◽  
Optimal Medium

Aim. Micropropagation of Jacob’s ladder Polemonium caeruleum L. and black salsify Scorzonera hispanica L., obtaining root culture and regenerated plants. Methods. In vitro plant cultivation, medium composition modification for micropropagation, inoculation of explants with agrobacterial strains. Results. In vitro cultures of Jacob’s ladder and black salsify have been obtained, the optimal medium composition has been determined for the effective plants multiplication, rooting and growth, root cultures and regenerated plants of studied species have been obtained. Conclusions. Obtained technology of in vitro culture establishment of P. caeruleum and S. hispanica can be used for plants microclonal propagation so as root culture and regenerated plants acquiring due to the agrobacterial transformation – for further studies of secondary metabolism of these plants. Keywords: P. caeruleum L., S. hispanica L., micropropagation, phytohormones, root culture.

scholarly journals Analysis of Genetic Variability Amongst Polyploid Genotypes of Pteris vittata L. From Various Geographic Locales of India

2021 ◽  
Vol 9 ◽  
Author(s):  
Jyoti Mathur ◽  
P. B. Khare ◽  
Apurva Panwar ◽  
S. A. Ranade
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Genomic Structure ◽  
Inter Simple Sequence Repeat ◽  
Pteris Vittata ◽  
Group Method ◽  
Bootstrap Analysis ◽  
Pair Group ◽  
Fern Species ◽  
Outgroup Taxon ◽  
Effective Multiplex Ratio

Pteris vittata L. is very common and a widely distributed species belongs to the family Pteridaceae. Various cytotypes from diploid to octaploid is available in this fern species. The present work has been carried out for genetic diversity in this fern both within and between the cytotypes. The molecular analysis at inter- as well as intra-species has been carried out with 57 accessions of P. vittata as well as of other species of Pteris with Microsorium punctatum considered as an out group taxon. For the present study 48 P. vittata (36 tetraploid and 12 pentaploid) and five of other species (four P. cretica, one P. pellucida, one P. tremula, one P. quadriaurita, and two P. ensiformis) accessions were used. The UPGMA (unweighted pair group method with arithmetic mean) dendrograms were generated for each method separately, as well as for all methods cumulatively, after a 1000 replicate bootstrap analysis. In order to determine the utility of each of the method, a comparative statistical assessment was done and marker index (MI), expected average heterozygosity, fraction of polymorphic loci and effective multiplex ratio (EMR) were calculated in case of each of the methods used in the present study. At the level of individual methods highest MI was obtained for directed amplification of minisatellites DNA (DAMD) method. Our findings of the present study concluded that out of the three methods Random Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeat (ISSR), and Directed Amplification of Minisatellite DNA (DAMD), DAMD was the best in term of polymorphism and heterozygosity as scores exhibited highest MI. The different accessions of P. vittata collected from different phytogeographical regions falls into six groups. Out of six clusters, one cluster is of pentaploid cytotype, four clusters are of tetraploid cytotype and one for outgroup taxon (M. punctatum). The result thus showed that within tetraploid, heterozygosity with variable genomic structure exists.

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