scholarly journals Silver nanoparticles induce genetic, biochemical, and phenotype variation in chrysanthemum

2020 ◽  
Vol 143 (2) ◽  
pp. 331-344
Author(s):  
Alicja Tymoszuk ◽  
Dariusz Kulus
Keyword(s):  
Silver Nanoparticles ◽  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Issr Markers ◽  
Phenotype Variation ◽  
Inter Simple Sequence Repeats ◽  
Tremendous Progress ◽  
Genetic Level ◽  
User Friendly

Abstract Despite the tremendous progress in breeding, novel and user-friendly techniques of plant improvement are desirable. The study aimed to analyze the usefulness of silver nanoparticles (AgNPs) in the breeding of chrysanthemum: one of the top ornamental plant species. In vitro regeneration of adventitious shoots from internodes of chrysanthemum ‘Lilac Wonder’ was induced on the modified Murashige and Skoog (MS) medium supplemented with 0.6 mg L−1 6-benzylaminopurine (BAP), 2 mg L−1 indole-3-acetic acid (IAA) and AgNPs at 0, 5, 10 and 20 ppm concentration. The efficiency of callogenesis and caulogenesis were analyzed after 10 weeks of culture. The concentration of chlorophylls, carotenoids, and phenolic compounds in shoots and calli were estimated. Plants obtained from 20 ppm AgNPs treatment were additionally analyzed on the genetic level using randomly amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers. In vitro rooted shoots were acclimatized in the glasshouse and subjected to biochemical and phenotype stability evaluation. AgNPs at the highest concentration (20 ppm) suppressed both callogenesis and caulogenesis in vitro. The concentration of metabolites in callus was stable, regardless of AgNPs treatment, except for carotenoids which production was enhanced by 20 ppm AgNPs. In contrast, the content of chlorophyll a and b in shoots varied depending on AgNPs treatment. Polymorphic loci were detected in 12 and 9 AgNPs-treated-plants by RAPD and ISSR markers, respectively (one of which was common to both marker systems). Rooting and acclimatization were fully successful in all experimental combinations. Phenotype alternations were detected in six plants; one from 10 ppm AgNPs treatment and five from 20 ppm treatment. They included variation in pigment content (anthocyanins and carotenoids) and/or inflorescence shape. Interestingly, only two plants revealed both genetic and phenotype polymorphisms. No genetic or phenotype variation was detected in the control plants. In conclusion, AgNPs can be used in chrysanthemum breeding.

scholarly journals In Vitro Regeneration of Coral Tree from Three Different Explants Using Thidiazuron

2019 ◽  
Vol 29 (6) ◽  
pp. 946-951 ◽  
Author(s):  
Saad B. Javed ◽  
Abdulrahman A. Alatar ◽  
Mohammad Anis ◽  
Mohamed A. El-Sheikh
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Issr Markers ◽  
Shoot Tip ◽  
Shoot Number ◽  
Efficient Alternative ◽  
Coral Tree ◽  
Hypocotyl Tissue ◽  
The Cost

The coral tree (Erythrina variegata) is a multipurpose horticultural plant with a plethora of medicinally important alkaloids. Regeneration via tissue culture can provide an efficient alternative to seed-grown plantlets and reduce the cost of the plant significantly. Thidiazuron (TDZ) is an efficient plant growth regulator and is effective in numerous species. However, the response to it varies with the type and position of the tissue on the plantlet treated. This study was carried out to ascertain the best tissue types for micropropagation of the coral tree using TDZ. Three tissue types (shoot tip, nodal, and hypocotyl), originating from different strata of the plantlet were evaluated. Adventitious shoots were observed in all three explants at the tested concentrations. However the quality and the shoot number varied significantly with the type of explant. Explants with a meristematic zone (shoot tip and nodal) were more responsive to the treatment compared with hypocotyl tissue lacking preexisting meristem. Nodal explants produced the maximum number of shoots (about eight) per explant after 4 weeks of culture, whereas shoot tips produced about only five shoots per explant at an equimolar concentration (1.5 µm). Approximately three shoots were observed in hypocotyl explants. Moreover, growth and rooting of the regenerated shoots was influenced by the origin of the explants. The molecular characterization of the regenerants using intersimple sequence repeat (ISSR) markers revealed genetic homogeneity among regenerants. An efficient micropropagation method for the coral tree is described.

scholarly journals Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 755
Author(s):  
Angela Ricci ◽  
Luca Capriotti ◽  
Bruno Mezzetti ◽  
Oriano Navacchi ◽  
Silvia Sabbadini
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Cultures ◽  
Efficient System ◽  
Regeneration Rate ◽  
Factors Affecting

In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.

scholarly journals Genotype-dependent mass somatic embryogenesis: a chance to recover extinct populations of Pulsatilla vulgaris Mill.

2021 ◽  
Author(s):  
Justyna Żabicka ◽  
Piotr Żabicki ◽  
Aneta Słomka ◽  
Monika Jędrzejczyk-Korycińska ◽  
Teresa Nowak ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Critical Point ◽  
Adventitious Shoots ◽  
Genetic Fidelity ◽  
Issr Markers ◽  
Molecular Techniques ◽  
Ex Situ ◽  
Experimental Conditions ◽  
Regenerated Plantlets

Abstract The paper presents a technique for micropropagation of endangered in Europe and extinct in Poland Pulsatilla vulgaris for ex situ conservation of the genetic resources. Genotype-dependent induction of somatic embryogenesis and rooting was revealed in series of two experiments (I and II) performed under the same experimental conditions. Shoot tips of seedlings were the best explants in both experiments and Murashige and Skoog (MS) medium supplemented with 0.25 or 0.5 mg L−1 BAP was suitable for induction of somatic embryos (SE) and adventitious shoots. Mass SE was obtained in experiment I after explants transfer on ½ MS (2% sucrose) + 0.45 mg L−1 B1 and extending culture to 2–3 months without passages. Rooting of adventitious shoots was a critical point. Out of seven rooting media used in experiment I, only two, ½ MS hormone free (2% sucrose) + 0.45 mg L−1 B1 or MS + 5 mg L−1 NAA + 3.76 mg L−1 B2 resulted in altogether 36.4% rooted shoots. In experiment II, somatic embryogenesis, rooting and acclimatization of adventitious shoots failed. Regenerated plantlets and seedlings converted from SE from experiment I were acclimatized to ex vitro conditions. Both genome size, determined by flow cytometry, and genetic diversity analyzed by ISSR markers, confirmed the compatibility of regenerants from experiment I with P. vulgaris initial seedlings and commercial cultivar. Regenerants obtained in experiment II differed genetically from the regenerants of experiment I and cultivar. Propagated in vitro tissues/organs (SE, adventitious shoots) of P. vulgaris could be a source of material for cryopreservation, artificial seed production and/or for acclimatization of regenerated plantlets and could be used for restoration of the extinct populations. Key Message The micropropagation technique via organogenesis and somatic embryogenesis of endangered in Europe pasqueflower was developed as a tool for species recovery. The critical point is that somatic embryogenesis is genotype-dependent, which affects the repeatability of the experiments and also imposes applying molecular techniques to confirm the genetic fidelity of the regenerants with the initial material and other genotypes.

scholarly journals In vitro Regeneration of Japanese Burdock (Arctium lappa L.) from Cotyledon and Leaf Derived Explants

2022 ◽  
Vol 31 (2) ◽  
pp. 123-134
Author(s):  
Mustafa Abul Kalam Azad ◽  
Md Arifuzzaman ◽  
Md Mobarok Hossain ◽  
Md Sohel Arman ◽  
Muhammad Nurul Amin
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Direct Shoot Regeneration ◽  
Indirect Organogenesis ◽  
Arctium Lappa ◽  
Sand Mixture ◽  
The Third ◽  
Rooting Medium

Considering the vegetable and medicinal values, a micropropagation protocol has been established for Japanese Burdock (Arctium lappa L.) by culturing the explants of cotyledon and leaf obtained from in vitro grown seedlings. Direct shoot regeneration was achieved from cotyledon and leaf explants on MS fortified with 4.0 μM BAP and 2.0 μM IBA or NAA after 5 weeks of culture. In addition, both the explants also formed callus from their cut margins within 6 weeks of cultivation on medium complemented with 6.0 μM BAP and 4.0 μM IBA or NAA. Adventitious shoots were also redeveloped through indirect organogenesis from the cotyledon and leaf-derived callus within 10 weeks of culture on MS containing 4.0 μM BAP and 2.0 μM IBA or NAA. The highest rate of shoot reproduction was attained at the third subculture, and more than 12.6 shoots were formed per callus clump. Within 4 weeks of transfer to the rooting medium on MS containing 6.0 μM IBA, the cultured micro-shoots produced highest 5.3 roots per cultured shoot. Rooted plantlets were successfully established on a soil-composed-sand mixture under natural condition with 93.3% survival rate Plant Tissue Cult. & Biotech. 31(2): 123-134, 2021 (December)

In vitro regeneration and assessment of genetic fidelity of acclimated plantlets by using ISSR markers in PPR-1 (Morus sp.): An economically important plant

2018 ◽  
Vol 241 ◽  
pp. 313-321 ◽  
Author(s):  
Gulab Khan Rohela ◽  
Phanikanth Jogam ◽  
Aftab Ahmad Shabnam ◽  
Pawan Shukla ◽  
Sadanandam Abbagani ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Genetic Fidelity ◽  
Issr Markers

scholarly journals In vitro Regeneration of the Medicinal Herb of Nilgiri shola, Acmella calva L. from Leaf Derived Callus

1970 ◽  
Vol 17 (2) ◽  
pp. 109-114 ◽  
Author(s):  
P. Senthilkumar ◽  
S. Paulsamy ◽  
K. K. Vijayakumar ◽  
K. Kalimuthu
Keyword(s):  
Key Words ◽  
Callus Culture ◽  
Callus Induction ◽  
Plant Tissue ◽  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Basal Medium ◽  
Root Initiation ◽  
Organogenic Callus

Callus culture was initiated from leaf discs on MS supplemented with BAP, NAA and Kn. The highest frequency (95%) of organogenic callus induction was observed in MS containing BAP at 3.0 mg/l and NAA at 0.3 mg/l. Development of adventitious shoots occurred when the calli were subcultured on MS supplemented with BAP alone at the rate of 3.0 mg/l (80%) and BAP with NAA at the rate of 3.0 and 0.3 mg/l, respectively (95%) and BAP with Kn at the rate of 3.0 and 0.3 mg/l, respectively (70%). The root initiation and the rate of growth were higher in the basal medium containing BAP at 3.0 and 2.5 mg/l. The plantlets were establish successfully in the hardening medium composed by coir pith and soil (1 : 1).Key words: Acmella calva, Leaf derived callus, Nilgiris, India  Plant Tissue Cult. & Biotech. 17(2): 109-114, 2007 DOI: 10.3329/ptcb.v17i2.2566

scholarly journals Assessment of Genetic Stability on In Vitro And Ex Vitro Plants of Ficus Carica Var. Black Jack Using Issr and Damd Markers

2021 ◽  
Author(s):  
Ankita Rajendra Parab ◽  
Chew Bee Lynn ◽  
Sreeramanan Subramaniam
Keyword(s):  
Stability Analysis ◽  
Genetic Stability ◽  
Similarity Index ◽  
Issr Markers ◽  
Ficus Carica ◽  
Ex Vitro ◽  
Regenerated Plants ◽  
Inter Simple Sequence Repeats ◽  
Dna Primers

Abstract In vitro propagation has been significant in producing a large number of genetically stable regenerated plants. Regenerated Ficus carica var. Black Jack plantlets were established using woody plant medium (WPM) supplemented with 20 µM 6-Benzylaminopurine (BAP) and 8 µM Indole-3-acetic acid (IAA) under different light treatments such as normal fluorescent white light (60 µmol.m− 2.s− 1), and four different LED spectra, white (400– 700nm), blue (440nm), red (660nm) and blue + red (440nm + 660nm). Genetic stability analysis was performed on the in vitro and ex vitro plants of Ficus carica var. Black Jack. Ten (10) primers of each ISSR and DAMD molecular marker were used to assess the genetic stability of the eight (8) samples of Ficus carica var. Black Jack, acquired over two years. The findings of this study revealed that inter simple sequence repeats (ISSR) and directed amplification of minisatellite DNA (DAMD) markers (DNA primers) are efficient in determining the polymorphism and monomorphism percentage among the in vitro and ex vitro samples of Ficus carica var. Black Jack. ISSR markers showed 97.87% of monomorphism whereas DAMD markers showed 100% monomorphism. Polymorphism of 2.13% was observed for the UBC840 ISSR – DNA primer which was negated under the genetic similarity index analysis for the eight samples. It is recommended that genetic stability analysis should be performed for long-term maintenance of micropropagated plants.

scholarly journals Comparison of Phytochemical and Antioxidant Activities in Micropropagated and Seed-derived Salvia miltiorrhiza Plants

HortScience ◽  
2018 ◽  
Vol 53 (7) ◽  
pp. 1038-1044 ◽  
Author(s):  
Bo-Ling Liu ◽  
Zhi-Bin Fan ◽  
Ze-Qun Liu ◽  
Xun-Hong Qiu ◽  
Yan-Hong Jiang
Keyword(s):  
Seed Germination ◽  
Antioxidant Activities ◽  
Salvia Miltiorrhiza ◽  
In Vitro Regeneration ◽  
Antioxidant Properties ◽  
Adventitious Shoots ◽  
Chemical Properties ◽  
Naphthalene Acetic Acid ◽  
Ms Medium

Salvia miltiorrhiza (commonly known in China as Danshen) is widely used in traditional Chinese medicine, and it is applied in the treatment of many diseases, particularly cardiovascular disease. Commercial propagation of Danshen is carried out either through seed germination or in vitro regeneration (micropropagation). However, it is not clear if the different propagation methods affect the chemical properties of the derived plants. In the present study, we first established a highly efficient tissue culture system for Danshen propagation. The addition of 1.0 mg·L−1 6-benzyladenine (BA) and 0.1 mg·L−1 α-naphthalene acetic acid (NAA) to Murashige and Skoog (MS) medium was optimal for inducing adventitious shoots; the highest rate of rooting was recorded on MS medium with 0.2 mg·L−1 NAA, on which the survival rate of transplanted plantlets was 95%. Next, we assessed antioxidant properties in the different tissues of plants of the same age, derived from micropropagation or seed germination, and measured tanshinone, total phenol, and total flavonoid contents. Our results showed that tissues of micropropagated plantlets had higher antioxidant activities than tissues of seed-derived plantlets; the micropropagated plantlets also had higher tanshinone contents in their roots. Thus, a rapid and efficient micropropagation system was established for Danshen, and it can be used for cultivating this plant to obtain therapeutic compounds.

scholarly journals In vitro regeneration and flowering of Portulaca grandiflora Hook

2019 ◽  
Vol 25 (4) ◽  
pp. 443-449 ◽  
Author(s):  
Carla Fernandes Cruz ◽  
Wolffe Ferreira dos Santos ◽  
Claudinei da Silva Souza ◽  
Marcelo Dias Machado ◽  
Ilio Fealho de Carvalho ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Ornamental Plant ◽  
Nodal Segments ◽  
Flower Buds ◽  
Floral Buds ◽  
Explant Source ◽  
Portulaca Grandiflora ◽  
Ornamental Species

Abstract P. grandiflora is a known ornamental plant with abundant flowering. The flowers exhibit varied coloration with distinct forms and simple folded petals and/or multiple. The objective of this work was to induce regeneration via organogenesis and in vitro flowering of P. grandiflora. Nodal segments of seedlings germinated in vitro were used as explant source for regeneration. Kinetin (KIN) and 6-Benzylaminopurine (BA) were used for the induction of organogenesis. The treatments supplemented with 1.0 and 1.5 mg L−1 BA induced the highest number of adventitious shoots with an average number of 7.0 (±1.55) e 5.4 (±0.83), respectively. The microcuttings obtained from regenerated shoots produced floral buds. The floral buds were located in the axillary and terminal regions of the microcuttings and developed in approximately 10 days of cultivation until the anthesis. The highest number of flower buds was observed in the presence of 0.75 mg L−1 of gibberellic acid. This study opens new perspectives for the establishment of biotechnological tools to be applied for this important ornamental species.

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