Clonal propagation and cryogenic storage of the medicinal plant Stevia rebaudiana

Shatnawi, Mohamad; Shibli, Rida A.; Abu-Romman, Saeid; Al-mazra’awi, Mohammad S.; Ajlouni, Z. I. Al; Shatanawi, Wasfı; Odeh, W.

doi:10.5424/sjar/20110901-021-10

Cited by 25 publications

(29 citation statements)

References 9 publications

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“…BAP promoted multiplication of Vitis vinifera and the rate of multiplication with BAP at 0.8 mg/L B (Shatnawi et al, 2011a). Similar results were obtained by Shatnawi et al (2011b) on Setivia rebuandia. BAP had inhibitory effect on shoot elongation (Tables 1 and 2).…”

Section: In Vitro Shoot Proliferationsupporting

confidence: 88%

“…In vitro conservation is based on conditions which permit minimal rates of growth, generally reduces the culture temperature or uses growth retardant (Marin and Duran-Vila, 1991;Orlikowska, 1992;Shibli et al, 1999;Sheyab et al, 2010;Shatnawi et al, 2011b). Sucrose functions as both a carbon/energy source and as an osmotic agent (Brown et al, 1979;Bonnier and Van Tuyl, 1997).…”

Section: Introductionmentioning

confidence: 99%

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Clonal propagation and medium-term conservation of Capparis spinosa: A medicinal plant

Al-Mahmood¹,

Shatnawi²,

Shibli³

et al. 2012

J. Med. Plants Res.

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Successful in vitro multiplication of Capparis spinosa was achieved on Murashige and Skoog (MS) medium supplemented with benzyl amino purine (BAP) at 0.8 mg/L. The highest shoot length (35.6 mm) was obtained with the use of 0.4 mg/L BAP and 0.2 mg/L 1-naphthaleneanacetic acid (NAA). Kinetin at 2.0 mg/L produced a multiplication rate of 9.7 microshoots per explants with an average shoot length of 21.3 mm. In vitro rooting was successfully achieved on MS media supplemented with different concentration of indole-3-butyric acid (IBA), indole acetic acid (IAA) or NAA at various concentrations. However, rooting did not occur in the absence of IBA, IAA or NAA. A total of 85% survival was achieved when rooted explants acclimatized ex vitro using a mixture of 1 perlite: 1 peat. In another experiment, in vitro C. spinosa were successfully stored without serious losses by using MS medium supplemented with an appropriate concentration of osmoticum (sucrose, sorbitol, mannitol or glucose) at various concentration (0, 3, 6, 9 or 12%). Two types of plant material were used (in vitro plantlets and in vitro plantlets without tips). The results obtained show that the two type of plant material could be successfully maintained in vitro and optimum treatments were identified for each plant material. Further studies are still needed on medium term conservation to enhance the survival percentages of different plant material type.

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Section: In Vitro Shoot Proliferationsupporting

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Clonal propagation and medium-term conservation of Capparis spinosa: A medicinal plant

Al-Mahmood¹,

Shatnawi²,

Shibli³

et al. 2012

J. Med. Plants Res.

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“…Also, Shatnawi et al (2011) reported that BA had better effect than Kinetin for increasing the number of shoot and length of Stevia.…”

Section: Discussionmentioning

confidence: 99%

Micropropagation of Stevia rebaudiana Betroni and assessment of genetic stability of in vitro regenerated plants using inter simple sequence repeat (ISSR) marker

Soliman¹,

Metwali²,

Almaghrabi³

2014

Plant Biotechnology

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Stevia rebaudiana Bertoni has become a new sweetener crop in different regions in the world. Stevia produces few seeds, therefore, micropropagation is a prevalent method to obtain sufficient amount of uniform plants. In the present study, in vitro propagation of Stevia was attempted through multiple shoot regeneration from nodal segments cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of 6-benzyladenine (BA), α-napthyalene acetic acid (NAA), indole acetic acids (IAA) and thidiazuran (TDZ). The maximum of number axillary shoots per explant (3.24) and highest shoot length (3.12 cm) were observed with MS medium supplemented with 1.0 mg l −1 BA+0.05 mg l −1 NAA and 2.0 mg l −1 BA+0.05 mg l −1 NAA, respectively. Roots were produced within two weeks and the highest percentage (98.72%) of root induction, the maximum number of roots (9.46 roots/shoot) and root length (9.87 cm) were recorded on MS medium fortified with 0.5 mg l −1 IAA and 1.5 mg l −1 IAA, respectively. The ex-vitro plantlets were successfully acclimatized with a survival rate of 70% at the hardening phase. Inter simple sequence repeat (ISSR) markers were used to analyze the genetic stability of micro-propagated and mother plants of Stevia. Four ISSR primers generated clear, distinct and reproducible bands. All ISSR profiles from micro-propagated plants were monomorphic and similar to mother plants, while low variation was induced in the next sub-culture. The results indicated that Stevia plantlets regenerated using micropropagation techniques standardized at our lab were genetically stable especially in the early sub-cultures.

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“…The seeds of Stevia show a very less vigour and low germination percentage [8]. Propagation by seeds does not allow the production of homogeneous populations, resulting in great variability of important features like sweetening levels and composition [9][10][11]. Vegetative propagation is too slow and limited by the lower number of individuals that can be obtained simultaneously from a single plant due to pathogen accumulation in the tissues [12][13][14].…”

Section: Introductionmentioning

confidence: 99%

Molecular Analysis of Genetic Fidelity in Micropropagated Plants of <i>Stevia rebaudiana</i> Bert. Using ISSR Marker

Lata¹,

Chandra²,

Techen³

et al. 2013

AJPS

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Inter-simple sequence repeat (ISSR) markers were used to evaluate the genetic fidelity of in vitro propagated and hardened plants of Stevia rebaudiana Bert. Nodal segments containing axillary buds were used as explant and inoculated on Murashige and Skoog's (MS) medium containing 3% (w/v) sucrose, 0.8% (w/v) agar supplemented with various concentrations of benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) ranging from 0.20 to 2.00 mg•L −1. Maximum multiple shoots (93%) were obtained in MS medium supplemented with 0.20 mg L −1 TDZ. The best in vitro root induction (87%) was achieved on half strength MS medium without any growth regulator. The rooted plantlets were successfully established in soil and grown to maturity at the survival rate of 96% in the indoor grow room. For ISSR analysis, total genomic DNA was extracted from 20 mg fresh leaves of mother and randomly selected in vitro propagated plants. Out of fifteen arbitrary primers tested, each produced clear and scorable amplification products ranged in size from about 216 bp in UBC 811 to 1917 bp in (GGGGT) 3 M with an average of 4.5 products per primer. A total of 45 bands (number of plantlets analyzed multiplied by number of bands with all primers) were generated by the ISSR method. All the ISSR profiles from micropropagated plants were monomorphic and comparable to mother plants, confirming the genetic stability among micropropagated plants and mother plant. Chemical analysis, using high-performance liquid chromatography (HPLC), was done to further confirm the existence of qualitative and quantitative differences in the major secondary metabolites (rebaudioside A, stevioside and steviolbioside) between the mother plant and in vitro propagated plants. Our results clearly show similar chemical profiles and insignificant differences in the major secondary metabolites between the two types of plants. These results suggest that the micropropagation protocol followed in this study is appropriate and applicable for clonal mass propagation of true-to-type elite Stevia rebaudiana plants.

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Clonal propagation and cryogenic storage of the medicinal plant Stevia rebaudiana

Cited by 25 publications

References 9 publications

Clonal propagation and medium-term conservation of Capparis spinosa: A medicinal plant

Clonal propagation and medium-term conservation of Capparis spinosa: A medicinal plant

Micropropagation of Stevia rebaudiana Betroni and assessment of genetic stability of in vitro regenerated plants using inter simple sequence repeat (ISSR) marker

Molecular Analysis of Genetic Fidelity in Micropropagated Plants of <i>Stevia rebaudiana</i> Bert. Using ISSR Marker

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Clonal propagation and cryogenic storage of the medicinal plant Stevia rebaudiana

Cited by 25 publications

References 9 publications

Clonal propagation and medium-term conservation of Capparis spinosa: A medicinal plant

Clonal propagation and medium-term conservation of Capparis spinosa: A medicinal plant

Micropropagation of Stevia rebaudiana Betroni and assessment of genetic stability of in vitro regenerated plants using inter simple sequence repeat (ISSR) marker

Molecular Analysis of Genetic Fidelity in Micropropagated Plants of &lt;i&gt;Stevia rebaudiana&lt;/i&gt; Bert. Using ISSR Marker

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Molecular Analysis of Genetic Fidelity in Micropropagated Plants of <i>Stevia rebaudiana</i> Bert. Using ISSR Marker