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 ◽  
...  

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.

2012 ◽  
Vol 3 (1) ◽  
pp. 6
Author(s):  
Beena D.B. ◽  
Rathore T.S.

Multiple shoots emerged from the nodal shoot segments of the field-grown candidate plus clump explants of <em>Bambusa pallida </em>Munro when cultured on Murashige and Skoog (MS) liquid medium with additives (ascorbic acid 50 mg/L + citric acid 25 mg/L + cysteine 25 mg/L) and combined use of α-naphthalene acetic acid (NAA) 1.34 μM + thiodiozuron 1.125 μM in a 2-week period. Further shoot multiplication was achieved in MS liquid medium with additives + NAA 1.34 μM + 6-benzylaminopurine 4.4 μM at 25±2°C and 33.78 μmol photons m-2 s-1 light illumination for a 12-h photoperiod. These shoots were rooted within four weeks in MS/2 basal salt medium with additives + 2% sucrose +1% glucose, and 0.6% agar by pulse treatment of shoots with indole 3 butyric acid 0.5 mg/mL for 30 min prior to inoculation. Rooted plants were successfully hardened in the mist chamber. Survival rate during hardening was more than 95%. Micropropagated plants achieved a height of 25-30 cm with 3-4 tillers (shoots) with miniature rhizome in a 4-month period. Genetic stability was observed in the micropropagated plants.


HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 579d-579
Author(s):  
Steve McCulloch

Briggs Nurseries, Inc. has used micropropagation as method of vegetative propagation for over 20 years. Genetic stability and uniformity of plants that are produced and sold is of the utmost concern to the commercial plant propagator. Genetic stability may be accomplished by ensuring that all shoots formed in vitro are of axillary origin and by reducing shoot proliferation rates through the use of lower cytokinin concentrations in the culture medium. Excision and removal of callus during transfer is also necessary to ensure that shoots develop from axillary buds. Various factors that may influence genetic variability and its frequency of in vitro derived plants will be discussed with an emphasis on how to reduce them. Three sources of variation with tissue culture derived plants will also be reviewed (Swartz, 1991): a) source plant variability, b) genetic changes in vitro, and c) epigenetic or physiological adaptation.


2014 ◽  
Vol 2014 ◽  
pp. 1-4 ◽  
Author(s):  
Dahanayake Nilanthi ◽  
Yue-Sheng Yang

Echinacea purpurea (purple coneflower) is being used for the preparation of more than 240 extracts, salves, and tinctures to help cure diseases like rabies, cold, and upper respiratory infections. Hence, efforts were made to develop a culture medium for successful in vitro culturing of cornflower and to regenerate buds and induce roots to enable mass propagation of selected clones. Of the three levels of sucrose tested as a supplement to MS media (Murashige and Skoog’s medium, 1962) 3% showed better rooting of buds and appeared morphologically normal and identical as compared to those grown at higher and lower concentrations (2 and 4%). The additives hydrolyzed lactabumin (0.0, 100, 300, and 900 mgL−1), peptone (0.0, 100, 300, and 900 mgL−1), and yeast (0.0, 100, 300, and 900 mgL−1) to media containing 0.3 mgL−1 BA (6-benzyladenine) and 0.01 mgL−1 NAA (naphthaleneacetic acid-plant growth regulators) has negatively influenced proliferation of shoots. The higher concentrations of the above have delayed the development of plantlets. Shoot multiplication was enhanced by coconut water with 2% being the best among 4 and 8% tested. Shoot organogenesis was not influenced by copper sulphate (0, 1.5, 3, 6, and 12 mgL−1) and silver nitrate (0.0, 0.5, 2.5, and 12.5 mgL−1) supplements and at higher concentrations of the above inhibited plant growth.


2017 ◽  
Vol 41 (6) ◽  
Author(s):  
Ricardo Gallo ◽  
Aloisio Xavier ◽  
Luciana Coelho de Moura ◽  
Brener de Almeida Oliveira ◽  
Heloisa Rocha do Nascimento ◽  
...  

ABSTRACT This study aims to evaluate the effect of IBA concentrations and microcuttings successive collections in the micropropagation of Eucalyptus grandis x E. urophylla and Eucalyptus urophylla x E. globulus clones. Clumps containing six to eight buds of clones established in vitro were transferred to a 250 mL glass flask in JADS semisolid medium. Successive collections were performed every 20 days for Eucalyptus grandis x E. urophylla clone and every 30 days for Eucalyptus urophylla x E. globulus clone. The following variables were evaluated under in vitro conditions: number of shoots > 0.5 cm, number of microcuttings > 2 cm, length of the longest microcutting, and shoots vigor. Under ex vitro conditions, in the greenhouse and shade house, the following variables were evaluated: seedling height, percentage of survival, stem diameter, percentage of root observed at the lower end of the tube, and seedling vigor. In full sun (ex vitro), the following variables were analyzed: seedling height, stem diameter, survival, number of roots, root volume, seedling vigor, and shoot and root dry matter. Good in vitro microcuttings productivity was observed over the successive collections. IBA levels were adjusted for each clone, ranging from 0.25 to 0.50 mg L-1 for Eucalyptus grandis x E. urophylla clone, and from 0.75 to 1.0 mg L-1 for Eucalyptus urophylla x E. globulus clone. IBA concentrations led to residual effects under ex vitro conditions, providing good rooting and survival for Eucalyptus grandis x E. urophylla and Eucalyptus urophylla x E. globulus clones at IBA concentrations between 0.25 and 0.50 mg L-1 and between 0.50 and 1.0 mg L-1, respectively.


Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 712
Author(s):  
Marzena Nowakowska ◽  
Žaklina Pavlović ◽  
Marcin Nowicki ◽  
Sarah L. Boggess ◽  
Robert N. Trigiano

Helianthus verticillatus (Asteraceae), whorled sunflower, is a perennial species restricted to a few locations in the Southeastern United States. Habitat loss has caused H. verticillatus to become rare, and since 2014, it has been federally listed as an endangered species. As a part of the recovery plan for the restoration and protection of H. verticillatus, an efficient micropropagation protocol based on axillary shoot proliferation was developed. Various concentrations of 6-benzylaminopurine (BAP; 0 to 4.44 µM) were examined for their morphogenetic potential in the regeneration of six genotypes of H. verticillatus from the nodal explants derived from greenhouse-grown plants. Both the BAP concentration and genotype had significant effects on the regeneration capacity of H. verticillatus. Although the induced buds were observed on ½-strength Murashige and Skoog medium without plant growth regulators, a higher rate of induction and bud development were achieved on media with either 0.88 or 2.22 µM BAP, regardless of the genotype. Successful rooting of the induced shoots was achieved within four weeks after the transfer from the induction medium to the fresh ½-strength MS medium, but the rooting efficiency was dependent on the plant’s genetic background. Regenerated plantlets, with well-developed shoots and roots, were acclimatized successfully to greenhouse conditions with a 97% survival rate. Simple sequence repeats (SSRs) markers were employed to assess the genetic uniformity of the micropropagated plants of H. verticillatus. No extraneous bands were detected between regenerants and their respective donor plants, confirming the genetic fidelity and stability of regenerated plants. To our knowledge, the protocol developed in this study is the first such report for this endangered species.


2013 ◽  
Vol 35 (8) ◽  
pp. 2467-2474 ◽  
Author(s):  
Sampath Kasthurirengan ◽  
Lifen Xie ◽  
Chun Hong Li ◽  
Yok King Fong ◽  
Yan Hong

2018 ◽  
Vol 42 (6) ◽  
Author(s):  
Denys Matheus Santana Costa Souza ◽  
Aloisio Xavier ◽  
Wagner Campos Otoni ◽  
Natane Amaral Miranda ◽  
Joane Helena Maggioni

ABSTRACT Micropropagation via axillary bud proliferation is recommended for rejuvenation or reinvigoration of selected clones, as well as for improving clonal seedlings rooting. The success of a micropropagation protocol depends on the in vitro introduction, since following phases, multiplication, elongation, and rooting can only take place once the aseptic crop with vegetative vigor has been established. This study aims to assess the effect of light on the in vitro introduction of hybrid clones of Corymbia torelliana x C. citriodora e Corymbia citriodora x C. torelliana by the micropropagation technique through proliferation by axillary buds. The mini-stumps, suppliers of explants for in vitro introduction, were conducted in semi-hydroclonal mini-clonal hedge. Nodal segments from three Corymbia torelliana x C. citriodora (TC01, TC02 e TC03) clones and one Corymbia citriodora x C. torelliana (CT01) clone were collected, disinfested and inoculated in JADS culture medium, in order to compare the effects of light quality from a dark/fluorescent lamp, a fluorescent lamp, and white and red/blue LEDs. At 30 days after inoculation, the following characteristics were evaluated: average contamination percentage, oxidation, non-reactive explants, shoot length and average number of shoots per explant greater than 0.5 cm. Gathered data showed that the use of red/blue LED light source obtained the best results in all assessed characteristics in the in vitro introduction.


2014 ◽  
Vol 62 ◽  
pp. 100-106 ◽  
Author(s):  
Mohammad Faisal ◽  
Abdulrahman A. Alatar ◽  
Ahmad K. Hegazy ◽  
Sulaiman A. Alharbi ◽  
Mohammad El-Sheikh ◽  
...  

2017 ◽  
Vol 4 (2) ◽  
pp. 17
Author(s):  
. Karyanti ◽  
Minda Kartini

Effect of Thidiazuron and Casein Hydrolysate on In Vitro Shoot Multiplication of Satoimo (Colocasia esculenta (L.) Schott var antiquorum)ABTRACTSatoimo (Colocasia esculenta (L.) Schott var antiquorum) is an alternative substitute of rice which has a big potential to be developed in Indonesia as an export commodity to Japan. Satoimo production needs to be increased to meet the demands for of the plant seeds. Plant propagation can be done using optimal media to stimulate the formation of shoots through, amongst others, the addition of thidiazuron (TDZ) and casein hydrolysate into the culture medium. This study aimed to determine the optimal concentration of TDZ and casein hydrolysate for in vitro multiplication of satoimo shoots. This research used RAL method with 2 factorials, namely the addition of TDZ at 0; 0.2; 0.6 mg/L concentrations, and of casein hydrolysate at 0; 150; 300; 450 mg/L concentrations. The results showed that the use of 0.6 mg/L TDZ and 150 mg/L casein hydrolysate resulted in the highest number of shoots, with the shoot average number of 6.9 per explant.Keywords: Casein hydrolysate, optimal medium, Satoimo, shoot multiplication, TDZ  ABTRAKSatoimo (Colocasia esculenta (L.) Schott var antiquorum) merupakan salah satu bahan alternatif pengganti beras yang memiliki peluang besar untuk dikembangkan di Indonesia, salah satunya sebagai komoditas ekspor ke negara Jepang. Produksi satoimo perlu ditingkatkan untuk memenuhi kebutuhan bibit tanaman tersebut. Perbanyakan tanaman dapat dilakukan menggunakan media yang optimal untuk merangsang pembentukan tunas, salah satunya dengan penambahan thidiazuron (TDZ) dan hidrolisat kasein pada media tanam. Penelitian ini bertujuan untuk mengetahui konsentrasi TDZ dan hidrolisat kasein yang optimal untuk perbanyakan tunas satoimo secara in vitro. Penelitian ini menggunakan metode RAL dengan 2 faktorial yaitu konsentrasi TDZ yang terdiri dari 0; 0,2; 0,6 mg/L dan konsentrasi hidrolisat kasein yang terdiri dari 0; 150; 300; 450 mg/L. Hasil penelitian menunjukkan bahwa pemberian TDZ 0,6 mg/L dan hidrolisat kasein 150 mg/L menghasilkan jumlah tunas tertinggi, dengan rata-rata tunas yang terbentuk 6,9 per eksplan.Kata kunci: Hidrolisat kasein, multiplikasi tunas, optimasi media, Satoimo, TDZ 


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