scholarly journals In vitro Regeneration and Agrobacterium-mediated Genetic Transformation of a Cultivated Potato Variety Using Marker Genes

2020 ◽  
Vol 30 (1) ◽  
pp. 149-160
Author(s):  
Sanjida Rahman Mollika ◽  
RH Sarker ◽  
MI Hoque
Keyword(s):  
Genetic Transformation ◽  
Potato Variety ◽  
Nodal Segments ◽  
Direct Organogenesis ◽  
Pcr Analysis ◽  
Bacterial Suspension ◽  
Marker Genes ◽  
Nodal Segment ◽  
Root Induction

Agrobacterium-mediated genetic transformation was carried out for Asterix (BARI Alu- 25), a popular potato (Solanum tuberosum L.) variety cultivated in Bangladesh. For Direct organogenesis of shoots the best response was noted when nodal segments and microtuber discs of Asterix along with Diamant - another popular potato variety were cultured on MS with 4.0 mg/l BAP and 1.0 mg/l IAA. MS without plant growth regulators was most effective for root induction from the excised regenerated shoots. Following optimum root development, the in vitro regenerated plantlets were successfully established in soil. Agrobacterium tumefaciens strain LBA4404/pBI121 containing GUS and nptII genes showed maximum transformation response in nodal segment with bacterial suspension having an optical density of 0.6 at 600 nm in Asterix variety. Moreover, 30 min incubation followed by 72 hrs co-cultivation was found most effective for transformation as has been determined by transient GUS histochemical assay. Transformed shoots were selected using MS with 4.0 mg/l BAP, 1.0 mg/l IAA, 0.5 mg/l GA3, 300 mg/l carbenicillin and 200 mg/l kanamycin. Stable integration of GUS and nptII genes were confirmed by PCR analysis using the genomic DNA isolated from transformed shoots. Plant Tissue Cult. & Biotech. 30(1): 149-160, 2020 (June)

scholarly journals In vitro Regeneration and Agrobacterium-mediated Genetic Transformation of Local Varieties of Mungbean (Vigna radiata (L). Wilczek)

2019 ◽  
Vol 29 (1) ◽  
pp. 81-97
Author(s):  
Sujay Kumar Bhajan ◽  
Setara Begum ◽  
Mohammad Nurul Islam ◽  
M Imdadul Hoque ◽  
Rakha Hari Sarker
Keyword(s):  
Genetic Transformation ◽  
Vigna Radiata ◽  
Zygotic Embryo ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Direct Organogenesis ◽  
Pcr Analysis ◽  
Root Induction ◽  
Half Strength

An efficient Agrobacterium-mediated transformation compatible in vitro regeneration protocol was developed for two important varieties of mungbean (Vigna radiata (L.) Wilczek) cultivated in Bangladesh, namely Binamoog-5 and BARI Mung-6. Two different zygotic embryo derived explants, such as cotyledonary node (CN) and cotyledon attached decapitated embryo (CADE) were used for direct organogenesis of shoot. MS supplemented with 4.0 μM BAP was found to be the best for the development of highest number of multiple shoots from CADE in both the varieties of mungbean. While in case CN the best shoot formation was achieved on MS containing 4.0 μM BAP and 0.5 μM NAA in both varieties. Half strength of MS with 2.0 μM IBA was found to be most effective for producing healthy root from regenerated shoots. Following root induction, the in vitro raised plantlets were successfully transplanted to soil for their establishment. Considering overall responses, genetic transformation efficiency was found to be better with CADE explant using Agrobacterium tumefaciens strain LBA4404 harboring the binary plasmid pBI121 conferring GUS and nptII genes. Different factors influencing transformation was optimized during this study. Selection of transformed shoots was carried out by gradually increasing the concentration of kanamycin and such transformed shoots were eventually selected using 200 mg/l kanamycin. Stable expression of the GUS gene was detected in various parts of regenerated transformed plantlets. Transformed shoots were rooted on half strength MS containing 2.0 μM IBA and 100 mg/l ticarcillin. Rooted transformed plantlets were successfully transferred to soil. Stable integration of GUS and nptII genes in the putative transformed shoots was confirmed through PCR analysis. Plant Tissue Cult. & Biotech. 29(1): 81-97, 2019 (June)

scholarly journals Genetic Transformation of a Local Tomato (Solanum lycopersicum L.) Variety of Bangladesh

2015 ◽  
Vol 25 (1) ◽  
pp. 87-97
Author(s):  
Pronabananda Das ◽  
Aneesa Ansari ◽  
Mohammad Nurul Islam ◽  
RH Sarker
Keyword(s):  
Genetic Transformation ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Bacterial Suspension ◽  
Marker Genes ◽  
Root Induction ◽  
Multiple Shoot Induction ◽  
Tomato Variety ◽  
Cotyledonary Leaf

An in vitro regeneration and Agrobacterium?mediated genetic transformation protocol was optimized for a local tomato variety, BARI Tomato?8 using cotyledonary leaf and hypocotyls explants. The explants were treated with various growth regulators in MS at different concentrations and combinations. Highest number of multiple shoot induction was observed from both the explants cultured in MS supplemented with 8.88 ?M BAP and 0.57 ?M IAA. Half strength of MS supplemented with 1.14 ?M IAA was found to be the best for root induction from excised shoots. Agrobacterium mediated genetic transformation was carried using pBI121 plasmid harboring ??glucuronidase (GUS) reporter and nptII selectable marker genes. Transient GUS assay confirmed that both the explants pre?cultured for two days showed best transformation efficiency in bacterial suspension having optical density (OD) of 0.8 (at 600 nm) for 15 min and co?cultivation period of 3 days. The shoots regenerated from transformed cotyledonary leaf explants survived at 200 mg/l kanamycin selection. The presence of expected amplicon corresponding to the GUS gene was confirmed by PCR. This protocol paves a way for developing disease resistant tomato variety using target gene/s.Plant Tissue Cult. & Biotech. 25(1): 87-97, 2015 (June)

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

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 High Frequency In vitro Regeneration of Gynura procumbens (Lour.) Merr.

2017 ◽  
Vol 27 (2) ◽  
pp. 207-216
Author(s):  
Tanjina Akhtar Banu ◽  
Barna Goswami ◽  
Shahina Akter ◽  
Mousona Islam ◽  
Tammana Tanjin ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Root Induction ◽  
Petiole Explants ◽  
Number Of Shoots

An efficient rapid in vitro regeneration protocol was described from nodal segment, leaf and petiole explants. MS medium supplemented with 1.0 mg/l BAP and 0.5 mg/l IAA was found best for the multiple shoot formation from nodal segments. In this combination 99% explants produced multiple shoots and the average number of shoots per explants was 20.1 ± 1.96. For petiole and leaf explants best response was observed on MS supplemented with 2.0 mg/l BAP, 1 mg/l IAA and 0.5 mg/l Kn. Petiole explants produced highest mean number of shoots/explant (22.9 ± 1.728) among the three explants when the explants were cultured on MS with 2.0 mg/l BAP, 1 mg/l IAA and 0.5 mg/l Kn. The highest frequency of root induction (100%) and mean number of roots/plantlets (11.75) were obtained on MS. The rooted plantlets were transferred for hardening following acclimatization and finally were successfully established in the field.Plant Tissue Cult. & Biotech. 27(2): 207-216, 2017 (December)

scholarly journals IN VITRO PROPAGATION OF SHOOTS AND CALLUS INDUCTION OF GYMNEMA SYLVESTRE R. BR. “AN IMPORTANT ANTI-DIABETIC PLANT”

2018 ◽  
Vol 10 (3) ◽  
pp. 60 ◽  
Author(s):  
Mohsina Syedy ◽  
Krishnendra Singh Nama
Keyword(s):  
Callus Induction ◽  
Research Work ◽  
Multiple Shoots ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Root Induction ◽  
Ms Medium ◽  
Petiole Explants ◽  
Leaf Petiole

Objective: The objective of this research was to establish and develop a protocol for the mass multiplication and callus induction of an anti-Diabetic plant-G. sylvestre R. Br.Methods: Sterilized explants (Nodal segment and leaf) were used for the initiation of culture. They were cultured on MS medium supplemented with a variety of PGRs (BAP, Kn, IBA, 2,4-D) individually or in combinations.Results: The induction of multiple shoots from nodal segments were highest in MS medium supplemented with 2.0 mg/l Kn and in BAP Maximum shoots were obtained on MS medium fortified with 1 mg/l BAP. For rooting different concentration of IBA were used and highest rooting was recorded on MS medium supplemented with 2.0 mg/l IBA. The rooted Plantlets were hardened initially in culture room conditions and then transferred to mist house. Leaf petiole explants were used for the purpose of callus induction. Best growth was observed in MS medium supplemented with 2,4-D. 1.0 mg/l 2,4-D+0.5 mg/l BAP, 1.0 mg/l 2,4-D+1.5 mg/l Kn.Conclusion: The results obtained in this research work clearly indicated that Kn is a better choice than BAP for the culture initiation. 2 mg/l IBA was proved best for root induction. For callus induction, 1 mg/l 2,4-D gave good results and when callus was sub-cultured on 2,4-D with BAP or Kn then 1.0 mg/l 2, 4-D+1.5 mg/l Kn proved best for mass propagation of callus.

scholarly journals Micropropagation of Salacia reticulata - an Endangered Medicinal Plant

2014 ◽  
Vol 23 (2) ◽  
pp. 221-229 ◽  
Author(s):  
G. Dhanasri ◽  
M. Srikanth Reddy ◽  
B. Naresh ◽  
Devi Cherku
Keyword(s):  
Medicinal Plant ◽  
High Performance ◽  
Antioxidant Properties ◽  
Shoot Multiplication ◽  
Nodal Segments ◽  
Direct Organogenesis ◽  
Root Induction ◽  
Over Exploitation ◽  
Salacia Reticulata

An efficient protocol of axillary bud proliferation and direct organogenesis has been developed for Salacia reticulata, a highly important medicinal plant. Over-exploitation for its antidiabetic and antioxidant properties concentrated in roots and stem has caused it to be endangered, thereby the need for its conservation. Propagation of S. reticulata in vitro is a promising way for its conservation. To develop the micropropagation protocol, the germplasm was screened for selection of a suitable ecotype with high content of mangiferin estimated with High Performance Liquid Chromatography technique. Nodal segments were cultured on MS supplemented with different growth regulators. The most efficient  shoot multiplication was obtained with the supplementation of BA and IAA (3.5 + 0.5 mg/l). Elongation of the micro-shoots was achieved by subculture every 20 days. The elongated micro-shoots were efficiently rooted in vitro on half strength MS supplemented with IBA. Plantlets were successfully established in the soil in 6 - 8 weeks and were morphologically similar to those of the source plant. The protocols developed presently for direct shoot regeneration and root-induction could be successfully applied for development of high quality planting stocks. D. O. I. http://dx.doi.org/10.3329/ptcb.v23i2.17523 Plant Tissue Cult. & Biotech. 23(2): 221-229, 2013  (December)

scholarly journals Enhanced Regeneration Through ex vitro Rooting and Agrobacterium-mediated Genetic Transformation of Eggplant (Solanum melongena L.)

2021 ◽  
Vol 31 (1) ◽  
pp. 97-108
Author(s):  
Sabina Yesmin ◽  
MI Hoque ◽  
RH Sarker
Keyword(s):  
Genetic Transformation ◽  
Solanum Melongena ◽  
Leaf Explants ◽  
Ex Vitro Rooting ◽  
Bacterial Suspension ◽  
Root Induction ◽  
Ex Vitro ◽  
Regeneration System ◽  
Cotyledonary Leaf

Regeneration of in vitro multiple shoots was achieved through organogenesis on MS supplemented with 2.0 mg/l BAP and 0.5 mg/l Kn from cotyledonary leaf explants of two local varieties of eggplant (Solanum melongena L.). Elongation of regenerated shoots was obtained on growth regulator free MS. In vitro root induction from excised regenerated shoots was less effective on MS with or without plant growth regulators. On the other hand regenerated shoots treated with 10 mM IBA for 5 min were found to be effective for ex vitro rooting in sterilized soil. Following sufficient development of roots, the ex vitro rooted plantlets were acclimatized in growth room condition, and were transferred to the field having 100% survival rate. The regeneration system developed was utilized for Agrobacterium-mediated genetic transformation using Agrobacterium tumefaciens strain LBA4404/pBI121 containing GUS and nptII genes. Adequate transformation response was obtained from cotyledonary leaf segments with bacterial suspension having an optical density of 0.50 at 600 nm with 30 min incubation followed by co-cultivation period of 72 hrs in Nayantara (BARI Begun-5) variety of eggplant. Selection of transformed shoots was carried out on MS supplemented with 2.0 mg/l BAP, 0.5 mg/l Kn, 300 mg/l carbenicillin and 100 mg/l kanamycin. Stable integration of GUS and nptII genes in Nayantara were confirmed through PCR analysis using the genomic DNA isolated from transformed shoots. Plant Tissue Cult. & Biotech. 31(1): 97-108, 2021 (June)

scholarly journals Targeting a Lipid Desaturation Enzyme, SCD1, Selectively Eliminates Colon Cancer Stem Cells through the Suppression of Wnt and NOTCH Signaling

Cells ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 106
Author(s):  
Yeongji Yu ◽  
Hyejin Kim ◽  
SeokGyeong Choi ◽  
JinSuh Yu ◽  
Joo Yeon Lee ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Notch Signaling ◽  
Cultured Cells ◽  
Pcr Analysis ◽  
Marker Genes ◽  
Dependent Manner ◽  
Lipid Desaturation ◽  
Novel Target

The elimination of the cancer stem cell (CSC) population may be required to achieve better outcomes of cancer therapy. We evaluated stearoyl-CoA desaturase 1 (SCD1) as a novel target for CSC-selective elimination in colon cancer. CSCs expressed more SCD1 than bulk cultured cells (BCCs), and blocking SCD1 expression or function revealed an essential role for SCD1 in the survival of CSCs, but not BCCs. The CSC potential selectively decreased after treatment with the SCD1 inhibitor in vitro and in vivo. The CSC-selective suppression was mediated through the induction of apoptosis. The mechanism leading to selective CSC death was investigated by performing a quantitative RT-PCR analysis of 14 CSC-specific signaling and marker genes after 24 and 48 h of treatment with two concentrations of an inhibitor. The decrease in the expression of Notch1 and AXIN2 preceded changes in the expression of all other genes, at 24 h of treatment in a dose-dependent manner, followed by the downregulation of most Wnt- and NOTCH-signaling genes. Collectively, we showed that not only Wnt but also NOTCH signaling is a primary target of suppression by SCD1 inhibition in CSCs, suggesting the possibility of targeting SCD1 against colon cancer in clinical settings.

scholarly journals Influence of Cytokinins in Combination with GA3on Shoot Multiplication and Elongation of Tea Clone Iran 100 (Camellia sinensis(L.) O. Kuntze)

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Reza Azadi Gonbad ◽  
Uma Rani Sinniah ◽  
Maheran Abdul Aziz ◽  
Rosfarizan Mohamad
Keyword(s):  
Camellia Sinensis ◽  
Woody Plants ◽  
Clonal Propagation ◽  
Shoot Multiplication ◽  
Shoot Elongation ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Liquid Form ◽  
Solid Media

The use ofin vitroculture has been accepted as an efficient technique for clonal propagation of many woody plants. In the present research, we report the results of a number of experiments aimed at optimizing micropropagation protocol for tea (Camellia sinensis(L.) O. Kuntze) (clone Iran 100) using nodal segments as the explant. The effect of different combinations and concentrations of plant growth regulators (PGR) (BAP, TDZ, GA3) on shoot multiplication and elongation was assessed. The influence of exposure to IBA in liquid form prior to transfer to solid media on rooting of tea microshoots was investigated. The results of this study showed that the best treatment for nodal segment multiplication in terms of the number of shoot per explant and shoot elongation was obtained using 3 mg/L BAP in combination with 0.5 mg/L GA3. TDZ was found to be inappropriate for multiplication of tea clone Iran 100 as it resulted in hyperhydricity especially at concentrations higher than 0.05 mg/L. Healthy shoots treated with 300 mg/L IBA for 30 min followed by transfer to 1/2 strength MS medium devoid of PGR resulted in 72.3% of shoots producing roots and upon transferring them to acclimatization chamber 65% survival was obtained prior to field transfer.

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