Genetic Stability of Micropropagated Ginger Derived from Axillary Bud through Cytophotometric and RAPD Analysis

2008 ◽  
Vol 63 (9-10) ◽  
pp. 747-754 ◽  
Author(s):  
Sujata Mohanty ◽  
Manoj K. Panda ◽  
Enketeswara Subudhi ◽  
Laxmikanta Acharya ◽  
Sanghamitra Nayak
Keyword(s):  
Dna Content ◽  
Genetic Stability ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Nuclear Dna Content ◽  
Basal Medium ◽  
Axillary Buds ◽  
Axillary Bud ◽  
Multiplication Rate

A protocol was developed for the in vitro propagation of ginger (Zingiber officinale) cv. Suprava using dormant axillary buds from unsprouted rhizomes. The dormant axillary buds embedded in the rhizome nodes were induced to sprout when cultured on MS medium supplemented with 6-benzyladenine (BA) alone (1 - 6 mg/l) or with a combination of BA (1 - 6 mg/l) and indole-3-acetic acid (IAA) (0.5, 1 mg/l). In vitro sprouted buds were transferred to the multiplication medium containing various combinations of auxins and cytokinins. MS basal medium supplemented with BA (1 mg/l), IAA (1 mg/l) and adenine sulfate (100 mg/l) was found optimum for the in vitro multiplication of shoots producing (8.2 ± 0.2) shoots from a single explant within 30 days of culture. The multiplication rate remained unchanged in subsequent subcultures. Rooting of shoots occurred in the same multiplication media. Upon transfer of the in vitro culture to ex vitro in pots, 96% of plants survived and established successfully under natural conditions. Tissue culture-raised plantlets of ginger could be conserved in vitro through subculturing at an interval of 4 months. The genetic stability of micropropagated clones was evaluated at regular intervals of 6 months up to 24 months in culture using cytophotometric estimation of 4C nuclear DNA content and random amplified polymorphic DNA (RAPD) analysis. Cytophotometric analysis revealed a unimodal distribution of the DNA content with a peak corresponding to the 4C value (23.1 pg), and RAPD analysis revealed monomorphic bands showing the absence of polymorphism in all fifty regenerants analyzed, thus confirming the genetic uniformity among in vitro grown somaclones of Z. officinale. This study is of commercial significance as axillary bud explants are available throughout the year for initiating a fresh culture of the elite ginger cv. Suprava to be used as a source of true-to-type disease-free planting material thereby minimizing the adverse effect of repeated subculturing from the same explant source.

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 Regeneration of Panax ginseng C.A. Meyer by Organogenesis and Nuclear DNA Analysis of Regenerants

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 628b-628
Author(s):  
Hak-Tae Lim ◽  
Haeng-Soon Lee ◽  
Tage Eriksson
Keyword(s):  
Panax Ginseng ◽  
Dna Content ◽  
Nuclear Dna ◽  
Adventitious Shoots ◽  
Flow Cytometric Analysis ◽  
Nuclear Dna Content ◽  
Basal Medium ◽  
Regeneration Ability ◽  
Half Strength

Plant regeneration ability of ginseng (Panax ginseng) via organogenesis was studied morphologically and anatomically. Compact callus was introduced from four different types of explants—leaf, petiole, flower stalk, and root—of in vitro-grown plantlets. Petioles were found to be the best material for callus induction. Calli induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (1.0 mg·L–1) and kinetin (0.1 mg·L–1) were conditioned for 2 weeks on the same medium. These calli differentiated into adventitious shoots when cultured on half-strength MS basal medium plus kinetin at 1.0 mg·L–1 and STS at 2.5 mg·L–1. An addition of GA3 (1.0 mg·L–1) and BA (1.0 mg·L–1) to MS basal medium, however, induced high-frequency in vitro flowering (86.1%) and multiple shoot budding, which affected the normal, complete development of plantlets. Plantlets with well-developed root systems were obtained 6 weeks after regenerated shoots had been transplanted to half-strength MS20 medium containing IBA at 0.25 mg·L–1. Nuclear DNA content was measured to check the stability of their ploidy level. Based on DNA flow cytometric analysis, all of the regenerants were typically diploids as were the mothers plants, indicating that nuclear DNA content remained stable during cell differentiation.

scholarly journals Rescue of a non-viable accession and rapd analysis of recovered plants of Arachis retusa

2004 ◽  
Vol 39 (2) ◽  
pp. 197-199 ◽  
Author(s):  
Rachel Fatima Gagliardi ◽  
Georgia Pereira Pacheco ◽  
Carlos Alberto Oliveira ◽  
Leonardo Alves Carneiro ◽  
José Francisco Montenegro Valls ◽  
...  
Keyword(s):  
Genetic Stability ◽  
Rapd Analysis ◽  
In Vitro Regeneration ◽  
Random Amplified Polymorphic Dna ◽  
Germplasm Preservation ◽  
Arachis Species ◽  
Embryo Axes ◽  
Genomic Regions ◽  
Regeneration Processes

In vitro regeneration of Arachis retusa was examined for the purpose of germplasm renewal and conservation. Random amplified polymorphic DNA (RAPD) fingerprinting was used to evaluate the genetic stability of plants derived from embryo axes and apical segments. Ten arbitrary decamer primers were screened and five of them were selected. Ninety genomic regions were evaluated, with an average of 18 loci per clone. All amplified segments were monomorphic. The results indicate that recovered plants are genetically stable at the assessed genomic regions and that both regeneration processes are suitable for in vitro germplasm preservation of Arachis species.

scholarly journals Genetic stability of wild pear (Pyrus pyraster, Burgsd) after cryopreservation by encapsulation dehydration

2008 ◽  
Vol 18 (2) ◽  
pp. 136 ◽  
Author(s):  
E. CONDELLO ◽  
M.A. PALOMBI ◽  
M.G. TONELLI
Keyword(s):  
Genetic Stability ◽  
Rapd Analysis ◽  
Leaf Shape ◽  
Alginate Beads ◽  
Mother Plant ◽  
Multiplication Rate ◽  
Fresh Weight ◽  
Base Pairs ◽  
Fresh Weight Basis ◽  
Pyrus Pyraster

Shoot tips of Pyrus pyraster were successfully cryopreserved by encapsulation-dehydration. Na--alginate beads each containing one shoot tip, dehydrated for 2 days in 0.75M sucrose and desiccated to 20% moisture content (fresh weight basis), gave 60% recovery after exposure to liquid nitrogen. Regenerated shoots showed no differences in length and leaf shape compared to the mother plant. Multiplication rate and rooting ability of cryopreserved shoots were lower than those of untreated controls after one subculture, but were completely restored following the third subculture. Fifteen cryopreserved lines derived from single buds were used for genetic analyses by RAPDs and SSRs, in comparison with the mother plant. In RAPD analysis, of a total of 24 primers, only 15 showed reproducible and well resolved bands and were further used. These primers produced a total of 66 fragments ranging from about 500 to 2500 base pair size. SSR (microsatellite) marker amplification was performed using 19 primers which produced 57 reproducible fragments. Microsatellites fragments ranged from 60 to 600 base pairs. Both RAPDs and SSRs did not reveal any polymorphism between cryopreserved lines and the original genotype, suggesting that cryopreservation, using encapsulation-dehydration, does not affect genetic stability of wild pear.;

In vitro propagation of the bay laurel (Laurus nobilis.L) in Morocco

2014 ◽  
Vol 4 (3) ◽  
pp. 96-103
Author(s):  
Abdelali Chourfi ◽  
Tajelmolk Alaoui ◽  
Ghizlane Echchgadda
Keyword(s):  
Seed Germination ◽  
In Vitro Culture ◽  
In Vitro Propagation ◽  
Aerial Part ◽  
Basal Medium ◽  
Axillary Buds ◽  
Laurus Nobilis

Laurus nobilis L. is among the species which are most threatened by massive degradation in Morocco. The multiplication by seed or by cuttings gives very low percentages of recovery that is insufficient to meet the demand of growing market. In vitro culture proves to be a tremendous asset to solve this problem. Our work has focused on the study of seed germination of this species and its multiplication from microcuttings. Finally, we studied the ac-climatization ability of the plantlets resulting from this germination. The study of the germination, via the further measurement of the length of the aerial part and the roots and the number of axillary buds for nine weeks, showed that the MS basal medium was more efficient than media 1/2M.S and WPM. Among the eight tested hormones, IAA yielded the best growth of the plantlets. Hormonal combination of NAA and kinetin resulted into a per-centage of the greatest success in reaching 67 % micropropagation. The study also revealed that the MS basal medium in the presence of the IAA plants can acclimate most easily in two types of substrates with improved development in the peat alone.

scholarly journals In Vitro Propagation of Three Moroccan Prickly Pear Cactus Opuntia and Plant Establishment in Soil

2013 ◽  
Vol 5 (1) ◽  
pp. 39-44 ◽  
Author(s):  
Aissam EL FINTI ◽  
Rachida EL BOULLANI ◽  
Naima AIT AABD ◽  
Fouad MSANDA ◽  
Mohammed A. SERGHINI ◽  
...  
Keyword(s):  
Basal Medium ◽  
Prickly Pear ◽  
Multiplication Rate ◽  
In Vitro Micropropagation ◽  
Prickly Pear Cactus ◽  
Half Strength ◽  
Micropropagated Plants ◽  
Large Production ◽  
Pear Cactus

Opuntia is one of the most widespread cacti, primarily due to their edible fruit and vegetable mass used as feed. The high demand for young plants of Opuntia made it necessary to find a rapid method of multiplication of the cactus, the safest method consisting in vitro micropropagation of species belonging to this genus. With aim of large production of plant material, a propagation system of three important prickly pear cactus cultivar (Opuntia ficus-indica) in Morocco was developed. Segments of healthy young cladode (containing one areole) were cultivated in Murashige and Skoog medium (MS) containing adenine sulfate (40 mg/1), monosodium phosphate (50 mg/l), sucrose (50 g/l), phytagel (0.3%) and benzyladenine (BA) at 22.2 μM, to start the process of micropropagation. In vitro-developed shoots from areoles were used as secondary explants to induce shoot development in the MS medium with 5 mg/l of BA. All of the three studied cultivars showed an important multiplication rate in this medium. ‘Sidi Ifni M’ (‘Moussa’) cultivar shows the greatest number of shoots followed by ‘Sidi Ifni A’ (‘Aissa’) and ‘Delahia’ 17.26, 14.12 and 12.13 respectively. Rooting of in vitro-generated shoots was achieved most efficiently on half-strength MS basal medium supplemented with 0.5 mg/l of indole-3-butyric acid (IBA) or IAA. Rooting frequencies were in the range from 95 to 100% and the highest mean number of root (19.1) was obtained with IBA for ‘Delahia’ cultivar. All micropropagated plants were transferred to greenhouse and all of them survived acclimatization process and showed good overall growth.

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