scholarly journals In vitro propagation of chia (Salvia hispanica L.) and assessment of genetic fidelity using random amplified polymorphic DNA and intersimple sequence repeat molecular markers

2019 ◽  
Vol 7 (1) ◽  
pp. 42-47 ◽  
HortScience ◽  
2014 ◽  
Vol 49 (1) ◽  
pp. 82-88 ◽  
Author(s):  
Samira Samarfard ◽  
Mihdzar A. Kadir ◽  
Saleh B. Kadzimin ◽  
Halimi M. Saud ◽  
Seyed Ali Ravanfar ◽  
...  

Protocorm-like bodies (PLBs) multiplication is one of the most preferable in vitro methods to increase the number of orchids that produce very few seeds or seeds that are not able to germinate. In the present study the effects of chitosan and thidiazuron (TDZ) combinations on multiplication, differentiation, and genetic stability of Phalaenopsis gigantea PLBs were investigated using different media. Initial PLBs were cultured in solid New Dogashima (ND) medium and Vacin and Went (VW) medium supplemented with different concentrations of chitosan (0, 5, 10, 15, 20, and 25 mg·L–1) and TDZ (0, 0.1, and 0.5 mg·L–1). The highest mean number of PLBs (353 PLBs) was observed in ND medium with 10 mg·L–1 chitosan and 0.1 mg·L–1 TDZ combination after 20 weeks of culture. Some PLBs differentiated into mature PLBs with a profusion of leaves on the apical region, and tiny plantlets started to develop after 10 weeks of culture. The highest mean number of shoots was observed in VW supplemented with 10 mg·L–1 chitosan and 0.5 mg·L–1 TDZ (16 shoots). Intersimple sequence repeat (ISSR) markers were used to determine the genetic fidelity among mother plant and PLBs obtained from each subculture stage of solid ND medium supplemented with 10 mg·L–1 chitosan and 0.1 mg·L–1 TDZ (the optimal treatment for PLB proliferation). Dissimilarity of 5% occurred between the mother plant and PLBs obtained after 16 weeks of culture. The range in the similarity coefficient varied from 0.80 to 1.0, and only 20% dissimilarity occurred between mother plant and PLBs after 20 weeks of culture.


Author(s):  
Asmaa Abdelsalam ◽  
Ehab Mahran ◽  
Kamal Chowdhury ◽  
Arezue Boroujerdi

Abstract Background Anarrhinum pubescens Fresen. (Plantaginaceae) is a rare plant, endemic to the Saint Catherine area, of South Sinai, Egypt. Earlier studies have reported the isolation of cytotoxic and anti-cholinesterase iridoid glucosides from the aerial parts of the plant. The present study aimed to investigate the chemical profiling of the wild plant shoots as well as establish efficient protocols for in vitro plant regeneration and proliferation with further assessment of the genetic stability of the in vitro regenerated plants. Results Twenty-seven metabolites have been identified in wild plant shoots using the Nuclear Magnetic Resonance (NMR) spectroscopy. The metabolites include alkaloids, amino acids, carbohydrates, organic acids, vitamins, and a phenol. In vitro propagation of the plant was carried out through nodal cutting-micropropagation and leaf segment-direct organogenesis. The best results were obtained when nodal cutting explants were cultured on Murashige and Skoog medium with Gamborg B5 vitamins supplemented with 6-benzylaminopurine (BAP) (1.0 mg/L) and naphthaleneacetic acid (NAA) (0.05 mg/L), which gave a shoot formation capacity of 100% and a mean number of shoots of 27.67 ± 1.4/explant. These shoots were successfully rooted and transferred to the greenhouse and the survival rate was 75%. Genetic fidelity evaluation of the micropropagated clones was carried out using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) molecular markers. Jaccard’s similarity coefficient indicated a similarity as high as 98% and 95% from RAPD and ISSR markers, respectively. Conclusions This study provides the chemical profiling of the aerial part of Anarrhinum pubescens. Moreover, in vitro regeneration through different tissue culture techniques has been established for mass propagation of the plant, and the genetic fidelity of the in vitro regenerated plants was confirmed as well. Our work on the in vitro propagation of A. pubescens will be helpful in ex situ conservation and identification of bioactive metabolites.


2017 ◽  
Vol 45 (2) ◽  
pp. 383-391 ◽  
Author(s):  
Constantinos SALIS ◽  
Ioannis E. PAPADAKIS ◽  
Spyridon KINTZIOS ◽  
Marianna HAGIDIMITRIOU

The behavior of six citrus rootstocks, Volkameriana, Citrumelo ‘Swingle’, Citrange ‘Carrizo’, Poncirus trifoliata ‘Serra’, Poncirus trifoliata ‘Rubidoux’ and Poncirus trifoliata ‘Flying Dragon’, in in vitro propagation was studied and compared for shoot proliferation and rooting. In addition, the genetic relationships among the rootstocks studied and other Citrus species, using the Inter-Simple Sequence Repeats (ISSR) molecular markers, were investigated. Nodal explants of three months old shoots were used in Murashige and Skoog medium supplemented with N6-benzyladenine (BA) for shoot proliferation and with naphthaleneacetic acid (NAA) for rooting. The rootstock Volkameriana showed a statistically significant higher number of shoots (1.81), shoot length (15.14 mm) and number of leaves per explant (5.81), while all three Poncirus trifoliata rootstocks showed the lowest numbers. The number of roots and root length per explant were evaluated at the end of the rooting phase. The rootstock ‘Swingle’ showed a higher number of roots per explant (4.2) followed by ‘Flying Dragon’ (3.93) and ‘Carrizo’ (3.23) rootstocks. The rootstocks ‘Swingle’ (140.8 mm), Volkameriana (148 mm) and ‘Flying Dragon’ (131.12 mm) had significantly higher root length per explant compared to ‘Carrizo’ (31 mm) and ‘Rubidoux’ (34.5 mm). The ISSR molecular marker technique used in the present study grouped successfully the different species, varieties and rootstocks studied, revealing their genetic variability. The genetic variability observed among the rootstocks ranged between 0.29 (Poncirus trifoliata ‘Serra’ and Citrumelo ‘Swingle’) and 0.60 (Volkameriana and Citrumelo ‘Swingle’). The response of the rootstocks studied in in vitro propagation however is not related to their genetic affinity.


Author(s):  
A. Z. Revutskaya ◽  
A. V. Holubenko ◽  
N. V. Nuzhyna ◽  
H. O. Rudik ◽  
N. Yu. Taran

Aim. Preparation of aseptic seedlings Salvia hispanica L., callus initiation in vitro and establishment of primary explants suitable for the callus production. Methods. Seeds are sprouted on our own modification of conventional methods. The non-hormonal Murashige-Skoog agarized nutrient medium was used as basic medium for the experiments. Parts of one-month seedlings (roots, hypocotyl, cotyledon leaves) were used as explants for the use of the colza. We added growth regulators (BAP, 2,4-D) in different concentration combinations into the nutrient medium for callus initiation. Statistical processing was performed in Microsoft Office Excel. Results. Aseptic S. hispanica seedlings have been obtained. The callus growth was initiated on all types of explants, the dependence of the callus intensity on the type of explants and the growth regulators content in the nutrient medium was established. Morphogenic callus and root-regenerants have been obtained. Conclusions. Hypocotyl was the most suitable primary explant for callus growth. Seedlings, leaves and roots showed low morphogenetic capacity. The nutrient medium with an elevated 2,4-D content was the most effective for initiation of callus genesis and proliferation of non-morphogenous callus. A high concentration of 2,4-D in the medium improves S. hispanica callus growth but suppresses its morphogenic ability.Keywords: Salvia hispanica (Chia), in vitro culture, callus.


2016 ◽  
Vol 29 (1) ◽  
pp. 481-494 ◽  
Author(s):  
Arockiam Sagina Rency ◽  
Lakkakula Satish ◽  
Subramani Pandian ◽  
Periyasamy Rathinapriya ◽  
Manikandan Ramesh

2017 ◽  
Vol 221 ◽  
pp. 1954-1961 ◽  
Author(s):  
Valeria Menga ◽  
Mariana Amato ◽  
Tim D. Phillips ◽  
Donato Angelino ◽  
Federico Morreale ◽  
...  

Biologia ◽  
2014 ◽  
Vol 69 (5) ◽  
Author(s):  
Ziba Bakhtiar ◽  
Mohammad Mirjalili ◽  
Ali Sonboli ◽  
Mahdi Farimani ◽  
Mahdi Ayyari

AbstractThymus persicus (Ronniger ex Rech. f.) Jalas is a valuable and endangered natural source of antitumor pentacyclic triterpenoids, i.e., betulinic acid, oleanolic acid and ursolic acid, which grows in northwest Iran. As the plant has a low propagation rate in nature, a suitable method for in vitro-propagation is needed. With the aim of identifying a suitable system for regenerating T. persicus via direct organogenesis, Murashige & Skoog (MS) medium supplemented with different plant growth regulators (PGRs) was tested. In vitro-grown shoot tips were exposed to the cytokinins 6-benzylaminopurine (BAP), kinetin (KN), and thidiazuron (TDZ), alone or in combination with the auxins 1-naphthalene-acetic acid (NAA), 2,4-dichlorophenoxyacetic (2,4-D), indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA). The highest shoot formation (7.1 ± 0.9) was obtained with a medium fortified with 8.9 μM BAP plus 2.7 μM NAA. Regenerated shoots were easily rooted on the different tested media, with the most abundant (16.6 ± 1.4) and strongest roots obtained on half-strength MS medium containing 2.5 μM IBA. The rooted plantlets were successfully acclimatized (76.6%) in a greenhouse before transference to natural conditions. Homogeneity and phytochemical productivity of the in vitro regenerated plantlets were confirmed by random amplified polymorphic DNA (RAPD) profiles and high performance liquid chromatography (HPLC), respectively.


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