scholarly journals Micropropagation of Environmental Stress Tolerant Local Potato (Solanum tuberosum L.) Varieties of Bangladesh

2014 ◽  
Vol 24 (1) ◽  
pp. 101-109 ◽  
Author(s):  
Fazlima Parveen ◽  
Mahmuda Khatun ◽  
Aparna Islam
Keyword(s):  
Shoot Apex ◽  
Large Scale ◽  
Solanum Tuberosum L ◽  
In Vitro Regeneration ◽  
Nodal Explants ◽  
Direct Regeneration ◽  
Nodal Segment ◽  
Scale Production ◽  
Potato Varieties

In vitro response of four explants namely, leaf, shoot apex, nodal and internodal, in three stress tolerant Bangladeshi potato varieties, viz. Zaubilati, Shadaguti and Challisha were tested. Of all the varieties, Shadaguti responded best for all the explants. Among the explants nodal segment responded best, followed by shoot apex. For all these explants and varieties, shoot regeneration response was tested in response to two cytokines, BAP and Kn. When compared between BAP and Kn supplementation, Kn treatment performed better than that of BAP for nodal and shoot apex, while opposite was observed for remaining explants. Interestingly, hormone free basal PROP medium was found to be best for nodal explants of both Zaubilati and Challisha varieties. While nodal explants of Shadaguti showed the best result in 0.5 mg/l Kn. Shoot apex  performed best in 0.5 mg/l Kn  for all the varieties. As an explant both internode and leaf did not perform well. Direct regeneration from these explants was found best in PROP + 0.2 mg/l GA3 + 0.5 mg/l IAA;  1.0 mg/l BAP, 1.5 mg/l BAP and 2.0 mg/l BAP for Zaubilai, Challisha and Shadaguti, respectively. For rooting, of the in vitro grown shoots half strength of PROP medium supplemented with 0.5 mg/l IBA was found to be best. Cent percent survival of transplanted plantlets was recorded. The successful protocol for in vitro regeneration was developed which can be used for large scale production of these abiotic stress tolerant potato varieties. Plant Tissue Cult. & Biotech. 24(1): 101-109, 2014 (June) D. O. I. http://dx.doi.org/10.3329/ptcb.v24i1.19218

In vitro mass propagation of Piper betle L

2019 ◽  
Vol 48 (3) ◽  
pp. 559-566
Author(s):  
Salim Khan ◽  
Barna Goswami ◽  
Shahina Akter ◽  
Mousona Islam ◽  
Afsana Huq Noon ◽  
...  
Keyword(s):  
Nodal Explants ◽  
Direct Regeneration ◽  
Leaf Segment ◽  
Piper Betle ◽  
Nodal Segment ◽  
Root Induction ◽  
Regeneration System ◽  
Petiole Explants ◽  
Best Response

An efficient in vitro regeneration system was developed for Piper betle L. through direct and indirect organogenesis from nodal segment, leaf segment and petiole explants. Highest direct regeneration was recorded when nodal explants were cultured on MS with 1.0 mg/l BAP and 1.0 mg/l Kn where 80% explants produced multiple shoots and the average number of shoots per explants were 3.20. Remarkable results on callus induction and shoot initiation were observed when the explants cultured on MS + 2.0 mg/l BAP + 0.5 mg/l Kn + 1.0 mg/l IAA. It was observed that nodal explants were showed best response on shoot/explants 13.2 ± 4.5 after 8 weeks of callus culture on MS medium with 0.5 mg/l BAP. The best response towards root induction was observed on half strength of MS with 0.25 mg/l IBA. The well rooted plants were successfully acclimatized and transferred to soil.

scholarly journals Agrobacterium-mediated Genetic Transformation for Local Cultivars of Potato (Solanum tuberosum L.) Using Marker Genes

1970 ◽  
Vol 20 (2) ◽  
pp. 145-155 ◽  
Author(s):  
Rita Sarah Borna ◽  
M. I. Hoque ◽  
R. H. Sarker
Keyword(s):  
Solanum Tuberosum ◽  
Genetic Transformation ◽  
Solanum Tuberosum L ◽  
In Vitro Regeneration ◽  
Direct Regeneration ◽  
Pcr Analysis ◽  
Marker Genes ◽  
Transformation Rate ◽  
Best Responses

Genetic transformation using nodal and internodal segments from three economically important potato (Solanum tuberosum L.) varieties namely, Diamant, Cardinal and Granola was conducted using an Agrobacterium tumefaciens strain LBA4404 harbouring binary plasmid pBI12 containing the GUS and nptII genes. Node and internodal segments were used for direct regeneration as well as regeneration with the intervention of callus. best responses were  obtained for direct regeneration of shoots when the explants were cultured on MS supplemented with 4.0 mg/l BAP +1.0 mg/l IAA, 1.5 mg/l BAP  + 0.5 mg/l IAA and 5.0 mg/l BAP +1.0 mg/l IAA in Diamant, Cardinal  and  Granola, respectively. In Diamant spontaneous in vitro microtuberization was obtained from these proliferated shoots. Further culturing of these in vitro grown green microtubers regenerated a large number of shoots on MS containing 4.0 mg/l BAP +1.0 mg/l IAA. By combining the best treatments, this protocol yielded an average transformation rate of 87% of treared explants. Stable expression of GUS gene was visualized in the various parts of transformed shoots through histochemical assay. Genomic DNA was isolated from transformed shoots and stable integration of the GUS and nptII genes was confirmed by PCR analysis.   Key words:  Potato, in vitro regeneration, transformation   D.O.I. 10.3329/ptcb.v20i2.6894   Plant Tissue Cult. & Biotech. 20(2): 145-155, 2010 (December)

scholarly journals IN VITRO REGENERATION OF POTATO (Solanum tuberosum L.)

2011 ◽  
Vol 24 (1) ◽  
pp. 07-14
Author(s):  
M. M. H. Molla ◽  
K. M. Nasiruddin ◽  
M. Al-Amin ◽  
M. S. Haque ◽  
D. Khanam
Keyword(s):  
Agricultural Research ◽  
Solanum Tuberosum L ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Zeatin Riboside ◽  
Direct Regeneration ◽  
Step Procedure ◽  
In Vitro Shoots ◽  
Nodal Cuttings

The experiment was conducted during the period from January to December, 2008 at the Laboratory of Biotechnology, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh. A two step procedure was followed for direct plant regeneration of potato. Murashige and Skoog (MS) basal medium supplemented with 0.5 mg l-1 IBA and 30 g sugar was used for in vitro potato plants development via nodal cuttings. Leaves and internodes of in vitro grown potato variety Asterix was cultured in instant MS basal medium supplemented with 0.01 mg l-1 IAA, 0.20 mg l-1 GA3, 44.0 g l-1 CaCl2, 20 g sucrose. Eight different concentrations of Zeatin riboside (ZR) viz., 0, 0.1, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0 mg l-1 along with above supplements (step II) were tested for in vitro direct regeneration of potato. MS medium supplemented with 4-5 mg l-1 of ZR performed better in respect of shoot induction from internodal explants. All the internodes produced shoots directly from the basal and apex region within 18-21 days and from the leaves within 21-28 days in 2 - 5 mg l-1 ZR. Maximum 43.20 and 7.25 shoots were recorded from each internode and leaf explant, respectively. In vitro shoots treated with 0.5 mg l-1 of IBA produced roots profusely within 21 days.DOI: http://dx.doi.org/10.3329/bjpbg.v24i1.16307

scholarly journals In Vitro Regeneration and ISSR-Based Genetic Fidelity Analysis of Orthosiphon stamineus Benth

Agronomy ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 778 ◽  
Author(s):  
Hanisah Ali ◽  
Izzah Farhanah Musa ◽  
Nurul Atikhah Abu Bakar ◽  
Saiful Anuar Karsani ◽  
Jamilah Syafawati Yaacob
Keyword(s):  
In Vitro Regeneration ◽  
Genetic Fidelity ◽  
Epileptic Seizures ◽  
Black Soil ◽  
Inter Simple Sequence Repeat ◽  
Issr Markers ◽  
Direct Regeneration ◽  
Nodal Segment ◽  
Orthosiphon Stamineus

Orthosiphon stamineus has been widely used as traditional remedy for various illnesses and diseases, such as cardiovascular diseases and epileptic seizures. In this study, direct regeneration through nodal segment of this species was attempted using Kinetin (6-Furfurylaminopurine) and IAA (indole-3-acetic acid). Optimum regeneration media was identified as MS media supplemented with 2.0 mg L−1 Kin plus 0.5 mg L−1 IAA. This yielded the highest number of shoots (5.57 ± 0.42) and leaves (20.53 ± 1.91) per explant. Acclimatization of the resulting in vitro regenerants was successful in all potting mixtures tested. However, potting mixture PF (1:1:1 ratio of black soil/red soil/compost) was identified as the best medium for acclimatization of this species, as it yielded 100% survival percentage after 90 days of acclimatization. Ten in vitro regenerants of O. stamineus were randomly collected after the third subculture and subjected to genetic variation analysis using inter-simple sequence repeat (ISSR) markers. Out of 20 ISSR markers tested, 10 working primers were observed to produce satisfactory amplification of bands, with an average of 7.11 bands per primer. A total of 610 bands were produced by the 10 primers. The percentage of polymorphism was observed to be very low, yielding only 7.32% polymorphism among all samples. Jaccard dissimilarity analysis was also conducted and very low genetic distance (about 0.1) was found among the in vitro regenerants and between the regenerants with the mother plant, thus ascertaining the clonal nature of the plantlets produced in this study.

scholarly journals Lentiviral Vectors for Delivery of Gene-Editing Systems Based on CRISPR/Cas: Current State and Perspectives

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1288
Author(s):  
Wendy Dong ◽  
Boris Kantor
Keyword(s):  
Large Scale ◽  
Lentiviral Vector ◽  
Clinical Applications ◽  
Scale Production ◽  
Base Editing ◽  
Epigenome Editing ◽  
Vector Systems ◽  
Dividing Cells

CRISPR/Cas technology has revolutionized the fields of the genome- and epigenome-editing by supplying unparalleled control over genomic sequences and expression. Lentiviral vector (LV) systems are one of the main delivery vehicles for the CRISPR/Cas systems due to (i) its ability to carry bulky and complex transgenes and (ii) sustain robust and long-term expression in a broad range of dividing and non-dividing cells in vitro and in vivo. It is thus reasonable that substantial effort has been allocated towards the development of the improved and optimized LV systems for effective and accurate gene-to-cell transfer of CRISPR/Cas tools. The main effort on that end has been put towards the improvement and optimization of the vector’s expression, development of integrase-deficient lentiviral vector (IDLV), aiming to minimize the risk of oncogenicity, toxicity, and pathogenicity, and enhancing manufacturing protocols for clinical applications required large-scale production. In this review, we will devote attention to (i) the basic biology of lentiviruses, and (ii) recent advances in the development of safer and more efficient CRISPR/Cas vector systems towards their use in preclinical and clinical applications. In addition, we will discuss in detail the recent progress in the repurposing of CRISPR/Cas systems related to base-editing and prime-editing applications.

Abstract 262: Derivation of Duchenne Muscular Dystrophy Disease Specific Cardiomyocytes From Patient Urine

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Xuan Guan ◽  
David L Mack ◽  
Claudia M Moreno ◽  
Fernando Santana ◽  
Charles E Murry ◽  
...  
Keyword(s):  
Stem Cells ◽  
Human Urine ◽  
Large Scale ◽  
Lentiviral Vector ◽  
Cellular Reprogramming ◽  
Urine Samples ◽  
Pcr Analysis ◽  
Scale Production ◽  
Mechanism Study

Introduction: Human somatic cells can be reprogrammed into primitive stem cells, termed induced pluripotent stem cells (iPSCs). These iPSCs can be extensively expanded in vitro and differentiated into multiple functional cell types, enabling faithful preservation of individual’s genotype and large scale production of disease targeted cellular components. These unique cellular reagents thus hold tremendous potential in disease mechanism study, drugs screening and cell replacement therapy. Due to the genetic mutation of the protein dystrophin, many DMD patients develop fatal cardiomyopathy with no effective treatment. The underlying pathogenesis has not been fully elucidated. Hypothesis: We tested the hypothesis that iPSCs could be generated from DMD patients’ urine samples and differentiated into cardiomyocytes, recapitulating the dystrophic phenotype. Methods: iPSCs generation was achieved by introducing a lentiviral vector expressing Oct4, Sox2, c-Myc and Klf4 into cells derived from patient’s (n=1) and healthy volunteers’ (n=3) urine. Cardiomyocytes were derived by sequentially treating iPSCs with GSK3 inhibitor CHIR99021 and Wnt inhibitor IWP4. Differentiated cardiomyocytes were subjected to calcium imaging, electrophysiology recording, Polymerase Chain Reaction (PCR) analysis, and immunostaining. Results: iPSCs were efficiently generated from human urine samples and further forced to differentiate into contracting cardiomyocytes. PCR analysis and immunostaining confirmed the expression of a panel of cardiac markers. Both normal and patient iPSC derived cardiomyocytes exhibited spontaneous and field stimulated calcium transients (up to 2Hz), as well as action potentials with ventricular-like and nodal-like characteristics. Anti-dystrophin antibodies stained normal iPSC-derived cardiomyocyte membranes but did not react against DMD iPSC-derived cardiomyocytes. Conclusions: Cardiomyocytes can be efficiently generated from human urine, through the cellular reprogramming technology. DMD cardiomyocytes retained the patient’s genetic information and manifested a dystrophin-null phenotype. Functional assessments are underway to determine differences that may exist between genotypes.

scholarly journals In vitro regeneration and induction of multiple shooting in Cicer arietinum L. using cotyledonary nodal explants

2015 ◽  
Vol 14 (13) ◽  
pp. 1129-1138 ◽  
Author(s):  
Prema Sunil Sruthi ◽  
Philip Robinson J ◽  
S KarthickBalan S ◽  
Anandhaprabhakaran M ◽  
Balakrishnan V
Keyword(s):  
Cicer Arietinum ◽  
In Vitro Regeneration ◽  
Nodal Explants ◽  
Multiple Shooting ◽  
Cicer Arietinum L

scholarly journals Assessment of In-vitro culture through Nodal explants of Dendrocalamus hamiltonii

2021 ◽  
Vol 2 (1) ◽  
pp. 130-133
Author(s):  
Abha Jha ◽  
◽  
Sunila Das ◽  
Keyword(s):  
Experimental Study ◽  
Growth Regulators ◽  
In Vitro Regeneration ◽  
Shoot Multiplication ◽  
Growth Hormones ◽  
Nodal Explants ◽  
Bud Break ◽  
Multiplication Rate ◽  
Dendrocalamus Hamiltonii

The present experimental study was aimed to overcome the traditional methods of propagation that limit the number of propagules by in-vitro regeneration through nodal explants of Dendrocalamus hamiltonii with a comparative study of growth regulators during the shooting and rooting process. Dendrocalamus hamiltonii is distributed from the Himalayas (Nepal) to the northern part of Burma. Collection of explants was done from different selected sites of CPTs. There was the use of HgCl2 and Ca (OCl)2 as sterilizing agents in different concentrations and its effect was visualized during the sprouting stage. Culm explants were cultured in a bottle containing White media (Wm) supplemented with BA and Kinetin for sprouting and IAA, IBA, NAA for rooting. There is also the use of IAA+IBA+NAA in combined form as a supplementary solution 0.1% HgCl2 treatment for 20-minute results into77.80% aseptic buds and 72% bud -break. Among the used growth-hormones, BA with concentration 0.25mg/l and 0.50mg/l respectively were appropriate for shoot-multiplication rate, 4.01±0.3 and 4.3±0.4 were ideal observation incorporation with BA (1.00mg/l) and BA (1.50mg/l) respectively. Maximum sprouting rate14.77±3.37with application of BA (2.00mg/l) and maximum shoot length4.3±0.4 is observed at BA (1.50mg/l). The applications of rooting hormone IAA+IBA+NAA in the concentration of 1.0 mg/l results in 72.5±0.3(rooting) and 11.1±0.3 (av. No. of the root).

scholarly journals Types and concentrations of cytokinins in the micropropagation of Anthurium maricense

2018 ◽  
Vol 12 (2) ◽  
pp. 117
Author(s):  
Cecília Moreira Serafim ◽  
Arlene Santisteban Campos ◽  
Priscila Bezerra Dos Santos Melo ◽  
Ana Cecília Ribeiro de Castro ◽  
Ana Cristina Portugal Pinto de Carvalho
Keyword(s):  
Light Intensity ◽  
Culture Medium ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Viable Option ◽  
Growth Room ◽  
In Vitro Induction

Faced with the demand for plants potted for their foliage, Anthurium maricense is seen as a viable option. However, most of the studies on obtaining micropropagated plantlets are for A. andraeanum, with nothing yet reported for A. maricense. The aim of this study therefore, was to evaluate the effect of four cytokinins in different concentrations, on the in vitro induction of shoots from nodal segments of A. maricense. The experimental design was completely randomised in a 4 x 4 factorial scheme, with four cytokinins (BAP, ZEA, CIN and 2iP) and 4 concentrations (0, 2.22, 4.44 and 6.66 μM), for a total of 16 treatments, with 6 replications of five test tubes, and using one nodal segment. Cultures were kept in a growth room at 25 ± 2°C, a photoperiod of 16 h and a light intensity of 30 μmolm-2 s-1 for 60 days. After this period, the number of shoots formed per node was evaluated. The addition of a cytokinin to the culture medium was determinant for the in vitro regeneration of shoots in A. maricense. The greatest estimated number of shoot formations in A. maricense were obtained in the culture media containing ZEA (3.87) and BAP (3.30), both at concentration of 6.66 μM.

Optimizing in vitro large scale production of giant reed (Arundo donax L.) by liquid medium culture

2014 ◽  
Vol 69 ◽  
pp. 21-27 ◽  
Author(s):  
Valeria Cavallaro ◽  
Cristina Patanè ◽  
Salvatore L. Cosentino ◽  
Isabella Di Silvestro ◽  
Venera Copani
Keyword(s):  
Liquid Medium ◽  
Large Scale ◽  
Arundo Donax ◽  
Giant Reed ◽  
Scale Production ◽  
Large Scale Production ◽  
Medium Culture ◽  
Arundo Donax L
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