scholarly journals Rapid in-vitro micropropagation of Bamboo (Dendrocalamus strictus) and its genetic fidelity testing using ISSR markers

2021 ◽  
Author(s):  
Shambhu Ram Khare ◽  
Pawankumar S. Kharate ◽  
Ritesh kumar Sahu ◽  
Zenu Jha
Keyword(s):  
Genetic Fidelity ◽  
Issr Markers ◽  
Mass Scale ◽  
Culture Technique ◽  
Nodal Segments ◽  
Tween 20 ◽  
Ms Medium ◽  
Dendrocalamus Strictus ◽  
Genetic Fidelity Testing

Bamboo is a versatile, arborescent, perennial and non-wood forest tree with tremendous commercial importance. For mass scale propagation of bamboo, the micropropagation is an effective way for producing elite, infection free and true-to-type planting material. Here, the nodal explants of Dendrocalamus strictus used to develop an effective protocol for micropropagation based on tissue culture technique. In this study, the sterilization treatment of 70% ethanol + Tween 20 + Bavistin  + Hgcl2 + PPM was successfully controlled the contamination up to 90 % as compared to other treatments. The shoots were initiated from nodal segments in MS medium supplemented with BAP (4 mg/l) and PPM (500µl/l). Shoot multiplication was found best with BAP (4 mg/l) and kinetin (2 mg/l) by using liquid MS medium. Whereas, rooting in solid MS medium has shown good results when supplemented with NAA (4mg/l). Healthy and disease-free plants were obtained after hardening under greenhouse conditions. Genetic fidelity testing by using ISSR markers reported that there was no variation in plantlets developed through micropropagation.

scholarly journals Efficient in vitro plant regeneration from leaf derived callus and genetic fidelity assessment of an endemic medicinal plant of Ranunculus wallichianus Wight & Arnn using RAPD and ISSR markers

2021 ◽  
Author(s):  
Srinivasan P ◽  
David Raja H ◽  
Tamilvanan R
Keyword(s):  
Callus Formation ◽  
Leaf Explants ◽  
Genetic Fidelity ◽  
Issr Markers ◽  
Ms Medium ◽  
Genetic Purity ◽  
Fidelity Assessment ◽  
Half Strength ◽  
Rapd And Issr

Abstract Ranunculus wallichianus is a medicinally important plant and an endemic species to Western Ghats of South India. An efficient and reliable indirect regeneration protocol system for R. wallichianus was developed from leaf explants in the present investigation. Leaf explants were cultured on both full-strength and half-strength MS (Murashige & Skoog) medium supplemented with different concentrations (1.0 mg L− 1 to 3.0 mg L− 1) of 2,4-D and NAA. Among the different concentrations tested, the highest percentage of yellowish green compact nodular callus formation was observed on half-strength MS medium with 2.0 mg L− 1 of 2, 4-D. Then, the in vitro raised organogenic callus was cultured on half strength MS medium containing various concentrations (1.0 mg L− 1 to 3.0 mg L− 1) of BA, KIN and TDZ with 0.5 mg L− 1 NAA and 10% CW for in vitro shoot regeneration. The highest percentage of regeneration response (97%) and maximum number of shoots formation (11.1 ± 0.13 shoots/culture with 9.2 ± 0.35 cm mean shoot length) were obtained from MS medium containing 2.5 mg L− 1 BA with 0.5 mg L− 1 NAA and 10% CW. The well elongated in vitro raised shoots were rooted in half strength MS medium with 2.5 mg L− 1 IBA + 250 mg L− 1 activated charcoal shows high frequency of root formation. The well rooted plantlets were successfully hardened and acclimatized with the survival rate of 94%. Clonal fidelity of in vitro raised plantlets was assessed by using DNA based RAPD and ISSR molecular markers. The total of 56 and 47 monomorphic bands were obtained from RAPD and ISSR markers respectively. This present in vitro propagation protocol system could be an effective for the conservation of R. wallichianus with their genetic purity and its further investigations.

scholarly journals Micropropagation and Assessment of Genetic Stability of In Vitro Raised Jojoba (Simmondsia chinensis Link.) Plants Using SCoT and ISSR Markers

2016 ◽  
Vol 25 (2) ◽  
pp. 165-179 ◽  
Author(s):  
SA Bekheet ◽  
AMM Gabr ◽  
AA Reda ◽  
MK El Bahr
Keyword(s):  
Genetic Fidelity ◽  
Issr Markers ◽  
Start Codon ◽  
Nodal Segments ◽  
Peat Moss ◽  
Adenine Sulphate ◽  
In Vitro Shoots ◽  
Micropropagated Plants ◽  
Number Of Shoots

This study is aimed at developing an efficient method for micropropagation of true-to-type jojoba plants. Nodal segments were used for in vitro shoots proliferation. Among three concentrations of BA used for multiplication, 1 mg/l BA gave the highest number of shoots. To enhance growth of shoots, combinations of BA and Kn were investigated. The greater value of shoot length and the maximum number of nodes were observed in the medium containing 1 mg/l BA + 1.5 mg/l Kn. Among different medium used to increase the rate of multiplication, the maximum number of shoots was recorded at ½ MS + Gamborg B5 (B5) vitamins + 2 mg/l Kn + 10 mg/l adenine sulphate (AS). Rooting was obtained upon supplementation of ½ Woody Plant Medium (WPM) with 1 mg/l IBA. Acclimation was achieved by transplanting rooted plantlets into pots containing peat moss and vermiculite. Start codon targeted (SCoT) and intersimple sequence repeat (ISSR) analyses were carried out to assess the genetic fidelity of micropropagated plantlets. All banding profiles of the two analyses from micropropagated plants were monomorphic and similar to the mother plant. Hence, this protocol can be employed for commercial micropropagation of jojoba without the risk of losing genetic stability.Plant Tissue Cult. & Biotech. 25(2): 165-179, 2015 (December)

scholarly journals Genetic Fidelity in Micropropagated Plantlets of Pimpinella tirupatiensis-an Endemic and Threatened Medicinal Plant Using RAPD and ISSR Markers

2016 ◽  
Vol 5 (06) ◽  
pp. 4661
Author(s):  
Srilakshmi A.* ◽  
Ugraiah A. ◽  
Gayatri C. M. ◽  
Rajanna L.
Keyword(s):  
Acetic Acid ◽  
Large Scale ◽  
Random Amplified Polymorphic Dna ◽  
Genetic Fidelity ◽  
Issr Markers ◽  
Scale Production ◽  
Ms Medium ◽  
Threatened Medicinal Plant ◽  
Issr Primers

Tissue cultured Pimpinella tirupatiensis plantlets were subjected to assessment of genetic stability considering the fact that associated in vitro stress might result in breakdown of control mechanism causing instability of the genome. We have used two DNA based molecular markers to assess the genetic fidelity of in vitro regenerated Pimpinella tirupatiensis through shoot tip from in vitro raised seedlings. The shoot tips upon transfer to MS medium containing different concentrations and combinations of 6- benzyl aminopurine (BAP) kinetin (KN), 2- isopentyl adenine (2- ip), α- napthalene acetic acid (NAA) and indole 3- acetic acid (IAA). The best morphogenic response was observed on MS medium fortified with BA (13.31 µM) and NAA (2.69 µM) which exhibited the highest regeneration frequency (90%), the maximum number of shoots/explants (6.50 ± 0.91) and shoot length (3.20 ± 0.20) within 5 weeks. Rooting was achieved within 15 days of shoot implantation on ½ strength MS media fortified with BAP (13.31 µM) and IBA (9.8 µM). The rooted plantlets were successfully acclimatized with 85% survival rate. Out of 20 RAPD and 3 ISSR primers screened, only 6 random amplified polymorphic DNA (RAPD) and all three inter simple sequence repeats (ISSR) primers produced clear reproducible and scorable bands. All banding profiles from micropropagated plants were monomorphic and similar to the mother plant indicating an absence of noticeable genetic variation in the regenerated plantlets. This study is of high significance as these could be commercially utilized for large scale production of true-to-type plantlets in Pimpinella tirupatiensis.

scholarly journals In vitro multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell. – a rare medicinal plant

2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Medicinal Plant ◽  
Large Scale ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ex Vitro ◽  
Culture Bottle ◽  
Hybanthus Enneaspermus ◽  
Half Strength

AbstractHybanthus enneaspermusis a rare medicinal plant. We defined a protocol for micropropagation,ex vitrorooting of cloned shoots and their acclimatization. Surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). Medium supplemented with 1.5 mg L−1BAP was found optimum for shoot induction from the explants and 6.4±0.69 shoots were regenerated from each node with 97% response. Shoots were further proliferated maximally (228±10.3 shoots per culture bottle with 7.5±0.43 cm length) on MS medium augmented with 1.0 mg L−1each of BAP and Kin within 4–5 weeks. The shoots were rootedin vitroon half strength MS medium containing 2.0 mg L−1indole-3 butyric acid (IBA). The cloned shoots were pulse-treated with 300 mg L–1 of IBA and cultured on soilrite® in a greenhouse. About 96% of the IBA-pulsed shoots rootedex vitroin soilrite®, each shoot producing 12.5±0.54 roots with 5.1±0.62 cm length. Theex vitrorooted plantlets showed a better rate of survival (92%) in a field study thanin vitrorooted plantlets (86%). A comparative foliar micromorphological study ofH. enneaspermuswas conducted to understand the micromorphological changes during plant developmental processes fromin vitrotoin vivoconditions in terms of variations in stomata, vein structures and spacing, and trichomes. This is the first report onex vitrorooting inH. enneaspermusand the protocol can be exploited for conservation and large-scale propagation of this rare and medicinally important plant.

scholarly journals Sterilisasi dan Induksi Daun Muda Durian (Durio zibethinus) Dalam Medium MS Dengan Penambahan Kinetin dan IAA Secara In Vitro

2021 ◽  
Vol 1 (1) ◽  
pp. 34-38
Author(s):  
Gatot Supangkat ◽  
Innaka Ageng Rineksane ◽  
Kurniawati Pamuji
Keyword(s):  
Vitamin C ◽  
Callus Formation ◽  
Tween 20 ◽  
Second Phase ◽  
Induction Phase ◽  
Ms Medium ◽  
Sterilization Method ◽  
Central Java ◽  
Durio Zibethinus

A research  to study the sterilization   method  and application   of Kinetin  and IAA to induce the Durian  young  leaf (Durio zibethinus) in MS  medium   was conducted in Balai Benih Induk Hortikultura in Salaman  Magelang  district  of Central  Java  started  on September  until December 2003. The Laboratory experiment   was arranged  in two phases,  which were  the optimation  phase of sterilization   and  induction   phase.  At  the  first  phase,  the  sterilization method  used  was  the modification   of Mulya  (2001) method.  The modification   use of sterilant,  vitamin  C antioxidant, Alcohol  70 %, Benlate, Agrept,  Tween-20  and Betadine  were done to obtain  effectiveness   of the sterilization.  Explants  planted  then in MS medium  for two weeks. Contamination   time, percentage of contamination   and viabilitas  (percentage of living explants)  were observed  then.  At the second phase,  the treatments were arranged  in a 3 x 3 factorial  completely   randomized   design  (CRD)  to observed  the influence  of Kinetin  and IAA combination.   The concentration   of Kinetin  observed were 2, 4, and 6 mg/I, where  as the IAA concentration   were 0.5,  1.0, and  1.5 mg/I. All treatments were  repeated  three  times,  with three samples  on each  replication.   The percentage   of browning explants, percentage  of contaminated   explants,  site of  contamination   and percentage of explants live were observed  at the end of incubation. The results  showed that sterilization  of Durian young leaves explants  with 1  g/l deterjent  for 15 minutes  then by 2 g/l Benlate  and Agrept  for 10 minutes,  then by 1  g/200 mg Vitamin C, then by Alcohol  70 % for 1  minute, then by 20% Clorox,  then by 2 drip of Tween-20  for 10 minute and then by Betadine  decreased  the contamination down to 50 %, and this kind of sterilization  was relatively better than  the other  kinds.  Application   of growth  regulators   were  not  able  to induce  explants growth,  but stimulated  callus formation  at the cutting surface though,  in the application  of Kinetin 4 mg/1 + IAA 0,5 mg/I, Kinetin 4 mg/1 + IAA  1,5 mg/1, Kinetin  6 mg/I+  IAA 0,5  mg/1 and Kinetin 6 mg/l+IAA   1,0 mg/I.

scholarly journals Micropropagation’s Complete Protocol of Red Araçá (Psidium cattleianum, Myrtaceae) from Germinated Seeds in vitro

2018 ◽  
Vol 10 (2) ◽  
pp. 234
Author(s):  
Cassio G. Freire ◽  
João P. P. Gardin ◽  
Cesar M. Baratto ◽  
Renato L. Vieira ◽  
Simone S. Werner
Keyword(s):  
Food Production ◽  
High Rate ◽  
Nodal Segments ◽  
Ms Medium ◽  
Brazilian Atlantic Forest ◽  
Psidium Cattleianum ◽  
Different Types ◽  
In Vitro Establishment ◽  
Germinated Seeds

Red Araçá’s (Psidium cattleianum) micropropagation processes have shown enormous potential both in terms of research and as a sustainable native resource to be used in the areas of food production, ecology, and pharmacology. Currently, however, despite that potential, research efforts involving this myrtaceae, native to the Brazilian Atlantic Forest, have been scarce. With that in mind, this study set out to establish micropropagation techniques that would allow the development of a feasible protocol to be used with Red Araçá, achieving its mircropropagation from in vitro germinated seeds. Different types of explants were tested for in vitro establishment. For the multiplication of nodal segments, different concentrations of BAP and IAA combinations were tested in an MS medium. Using the same medium, different concentrations of ampicillin were applied in order to determine its influence on the decontamination of the apical segments. The BAP and IAA combinations were also used to test their effects on the in vitro explants’ development and rooting. During pre-acclimatization, survival of in vitro rooted plants was tested in a nebulizer chamber, using a commercial substrate and that same substrate mixed with washed sand (1:1). In essence, it was indeed possible to develop a complete protocol for the micropropagation of the Red Araçá from seedlings obtained by in vitro germination. The in vitro introduction of the Red Araçá was rather efficient, independently of the type of explants used. As the BAP and IAA concentrations increased, so did the in vitro seedlings’ development (7 leaves explant-1) and rooting (67%). Additionally, the in vitro rooted plants exhibited a high rate of survival (80%) in the pre-acclimatization phase, independently of the substrate used.

scholarly journals In vitro regeneration protocol of Rauvolfia serpentina L.

2018 ◽  
Vol 53 (2) ◽  
pp. 133-138 ◽  
Author(s):  
S Khan ◽  
TA Banu ◽  
S Akter ◽  
B Goswami ◽  
M Islam ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Root Induction ◽  
Ms Medium ◽  
Regeneration System ◽  
Rauvolfia Serpentina ◽  
Number Of Shoots

An efficient in vitro regeneration system was developed for Rauvolfia serpentina L. through direct and indirect organogenesis from nodal and leaf explants. Among the different growth regulators, MS medium supplemented with 2.0 mg/l BAP, 0.5mg/l IAA and 0.02mg/l NAA found best for the multiple shoot formation from nodal segments. In this combination 98% explants produced multiple shoots and the average number of shoots per explants is 13∙4. The frequency of callus induction and multiple shoot induction from leaves was highest 88% in MS medium supplemented with 2.0 mg/l BAP, where mean number of shoots/explants was 12.5. The highest frequency of root induction (80%) and mean number of roots/plantlets (10) were obtained on half strength of MS medium containing 0.2 mg/l IBA. The rooted plantlets were transferred for hardening following acclimatization and finally were successfully established in the field.Bangladesh J. Sci. Ind. Res.53(2), 133-138, 2018

scholarly journals Regeneration of Plants from Apical Meristem Tips and Nodal Segments of Arachis pintoi

2003 ◽  
Vol 30 (2) ◽  
pp. 75-79 ◽  
Author(s):  
H. Y. Rey ◽  
L. A. Mroginski
Keyword(s):  
Apical Meristem ◽  
In Vitro Regeneration ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Regeneration Potential ◽  
Arachis Pintoi ◽  
Solid Media ◽  
One Step

Abstract The in vitro regeneration potential of shoot apical tips (2 to 3 mm in length), meristems (0.3 to 0.5 mm in length), and nodal segments (4 to 7 mm long with an axillary bud) of diploid (2n = 2x = 20) and triploid (2n = 3x = 30) cytotypes of Arachis pintoi was evaluated. Explants were cultured on MS medium supplemented with different concentrations and combinations of naphthaleneacetic acid (NAA) and benzyladenine (BA). In one experiment the effect of gibberellic acid was tested. The cultures were done in liquid and solid media. Plant regeneration can be readily achieved from all explants in one step of 30 d culture on MS + 0.01 mg/L each of NAA and BA or two steps consisting of 1) shoots regeneration through culture of explants on MS + 0.01 mg/L each of NAA and BA, and 2) induction of rooting in regenerated shoots by reculture on MS + 0.01 mg/L NAA. The plantlets were successfully transferred to pots in a greenhouse.

scholarly journals In vitro study of Tinospora cordifolia (Willd.) Miers (Menispermaceae)

2010 ◽  
Vol 6 ◽  
pp. 103-105 ◽  
Author(s):  
Aditi Singh ◽  
Saroj K Sah ◽  
Aunji Pradhan ◽  
Sabari Rajbahak ◽  
Niran Maharajan
Keyword(s):  
Medicinal Plant ◽  
Callus Induction ◽  
Shoot Growth ◽  
Tinospora Cordifolia ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Nodal Segment ◽  
Root Induction ◽  
Ms Medium

In vitro study was carried out in an important medicinal plant Tinospora cordifolia (Willd.) Miers belonging to the family: Menispermaceae. Vegetative parts such as stem, leaf and nodal explants were excised from an elite in vivo grown mature plant and thereafter cultured on Murashige-Skoog (MS) medium supplemented with different hormonal concentrations for callus induction and organogenesis. Callus formation occurred from nodal segments, leaf and inter-node explants when planted on different combinations of hormones. Tinospora cordifolia showed response for in vitro shoot growth from the nodal segment. The best shoot growth was observed on MS medium supplemented with kinetin (1.5 mg/l). Similarly, the best result for root induction was obtained on MS medium supplemented with 6-benzylaminopurine (1.0 mg/l) and naphthaleneacetic acid (2.5 mg/l). Key-words: callus induction; explants; medicinal plant; MS medium; tissue culture.DOI: 10.3126/botor.v6i0.2918 Botanica Orientalis - Journal of Plant Science (2009) 6: 103-105

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