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 Regeneration and Agrobacterium-mediated Genetic Transformation of Tomato (Lycopersicon esculentum Mill.)

1970 ◽  
Vol 19 (1) ◽  
pp. 101-111 ◽  
Author(s):  
Rakha Hari Sarker ◽  
Khaleda Islam ◽  
M.I. Hoque
Keyword(s):  
Lycopersicon Esculentum ◽  
Genetic Transformation ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Multiple Shoot ◽  
Root Induction ◽  
Cotyledonary Leaf ◽  
Lycopersicon Esculentum Mill

Agrobacterium-mediated genetic transformation system has been developed for two tomato (Lycopersicon esculentum Mill.) varieties, namely Pusa Ruby (PR) and BARI Tomato-3 (BT-3). Prior to the establishment of transformation protocol cotyledonary leaf explants from the two varieties were cultured to obtain genotype independent in vitro regeneration. Healthy multiple shoot regeneration was obtained from the cut ends of cotyledonary leaf segments for both the varieties on MS containing 1.0 mg/l BAP and 0.1 mg/l IAA. The maximum root induction from the regenerated shoots was achieved on half the strength of MS medium supplemented with 0.2 mg/l IAA. The in vitro grown plantlets were successfully transplanted into soil where they flowered and produced fruits identical to those developed by control plants. Transformation ability of cotyledonary leaf explants was tested with Agrobacterium tumefaciens strain LBA4404 harboring binary plasmid pBI121, containing GUS and npt II genes. Transformed cotyledonary leaf explants were found to produce multiple shoots on MS containing 1.0 mg/l BAP and 0.1 mg/l IAA. Selection of the transformed shoots was carried out by gradually increasing the concentration of kanamycin to 200 mg/l since kanamycin resistant gene was used for transformation experiments. Shoots that survived under selection pressure were subjected to rooting. Transformed rooted plantlets were transferred to soil. Stable expression of GUS gene was detected in the various tissues from putatively transformed plantlets using GUS histochemical assay.  Key words: In vitro regeneration, transformation, tomato D.O.I. 10.3329/ptcb.v19i1.5004 Plant Tissue Cult. & Biotech. 19(1): 101-111, 2009 (June)

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 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 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 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 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 Agrobacterium?mediated Genetic Transformation of Two Varieties of Brassica juncea (L.) Using Marker Genes

2016 ◽  
Vol 26 (1) ◽  
pp. 55-65 ◽  
Author(s):  
Shirin Akter ◽  
Sanjida Rahman Mollika ◽  
RH Sarker ◽  
M Imdadul Hoque
Keyword(s):  
Genetic Transformation ◽  
Brassica Juncea ◽  
Bacterial Suspension ◽  
Marker Genes ◽  
Positive Interaction ◽  
Agrobacterium Strain ◽  
Local Varieties ◽  
Cotyledonary Leaf ◽  
Gus Histochemical Assay ◽  
Binary Plasmid

Protocol for Agrobacterium?mediated genetic transformation using hypocotyl and cotyledonary leaf with petiole from two local varieties of Brassica juncea was established by optimizing various factors influencing transformation. GUS histochemical assay revealed that the cotyledonary leaf with petiole and hypocotyl explants had positive interaction with the Agrobacterium strain LBA4404 containing the binary plasmid pBI121 which has marker genes like, GUS and nptII. Maximum transformation was obtained with bacterial suspension having an optical density of 0.8 at 600 nm, 30 min of incubation and 72 hours of co?cultivation. The transient and stable integration of the marker genes were confirmed through histochemical GUS assay, as well as PCR analysis.Plant Tissue Cult. & Biotech. 26(1): 55-65, 2016 (June)

scholarly journals In vitro regeneration and molecular characterization of some varieties of Lycopersicon esculentum Mill. in Bangladesh

2019 ◽  
Vol 54 (2) ◽  
pp. 117-124
Author(s):  
M Billah ◽  
TA Banu ◽  
M Islam ◽  
NA Banu ◽  
S Khan ◽  
...  
Keyword(s):  
Lycopersicon Esculentum ◽  
Molecular Characterization ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Cotyledonary Node ◽  
Cotyledonary Leaf ◽  
Lycopersicon Esculentum Mill ◽  
Regeneration Response

An efficient regeneration protocol was established for two varieties (BARI tomato-9 and BARI tomato-15) of tomato (Lycopersicon esculentum Mill.) using three explants namely cotyledonary node, cotyledonary leaf and hypocotyls. Among the three explants, maximum number of shoots was produced from cotyledonary leaf explants of BARI tomato-15 on MS with 2.0 mg/l BAP and 0.5 mg/l IAA. In this combination of BAP and IAA 86%, on an average, cotyledonary leaf explants showed regeneration response 14.12 shoots/explants. Explants from hypocotyl showed best results in MS medium with 2.0 mg/l BAP and 0.2 mg/l IAA in both the varieties. In case of cotyledonary node, BARI tomato-15 showed 6.0 shoot/explant on MS with 2.0 mg/l BAP and 1.0 mg/l IAA. Molecular characterization of total ten varieties of tomato in Bangladesh was done by using six arbitrary oligonucleotide RAPD primers. A total of 140 bands were produced where the highest genetic distance (0.6769) was found between BARI tomato-3 and Mintoo tomato and lowest distance (0.1035) was observed between BARI tomato-7 and BARI tomato-8. This result will be useful for designing future breeding programs. Bangladesh J. Sci. Ind. Res.54(2), 117-124, 2019

scholarly journals IN VITRO DIRECT MULTIPLE SHOOT INDUCTION FROM LEAF EXPLANTS OF SOLANUM PUBESCENS WILLD

2016 ◽  
Vol 4 (12SE) ◽  
pp. 23-28
Author(s):  
Ayyadurai V ◽  
Ramar K
Keyword(s):  
Leaf Explants ◽  
Multiple Shoots ◽  
Multiple Shoot ◽  
Root Induction ◽  
Ms Medium ◽  
Multiple Shoot Induction ◽  
Half Strength ◽  
Multiple Shoot Regeneration ◽  
Solanum Pubescens

Efficientin Vitro direct multiple shoot regeneration from Solanum pubescens was achieved from leaf explants on MS medium Sublimated with B5 vitamins and different concentrations and different combinations of PGRs like BAP, NAA and GA3. The maximum numbers of multiple shoots were achieved from leaf explants on 3.0 mg/l BAP + 1.0mg/l GA3. The regenerated shoots were transferred in to half strength MS medium fortified with IBA for root induction. Rooted plantlets were successfully acclimatized. This new and transfer into the field Conditions. Standardized and reproducible protocol useful the mass propagation of Solanum pubescens.

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