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 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 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 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 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 Establishment of in vitro regeneration protocol for Adhatoda vasica Nees

2016 ◽  
Vol 51 (1) ◽  
pp. 75-80 ◽  
Author(s):  
S Khan ◽  
S Akter ◽  
A Habib ◽  
TA Banu ◽  
M Islam ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Garden Soil ◽  
Root Induction ◽  
Ms Medium ◽  
Adhatoda Vasica

An in vitro regeneration protocol of Adhatoda vasica has been developed using excised nodal segments and juvenile leaves for multiple shoots regeneration directly or through callus induction. Explants were cultured on MS medium with different concentrations of IAA, NAA, BAP, GA3 and Kn singly or in combinations. MS medium supplemented with BAP (10.0 mg/l) was found best for multiple shoot formation, in which 93.33% explants produced multiple shoots. After two months, maximum number of multiple shoots were 10.6 ± 1.82, highest length of plantlets was 5.2 ± 2.20 cm. 100% calli formation were observed on MS medium supplemented with IAA (0.05 mg/l) + NAA (0.05 mg/l) + BAP (1.0 mg/l). Callus initiation started after 14 days and gave light green colored callus. Best callus mediated shoot regeneration was found on MS+10.0 mg/l BAP medium. Root induction of in vitro raised shoots was best on ½ MS + IBA (1.0 mg/l). Well rooted plantlets were transferred to plastic pots containing garden soil and compost in a ratio of 2:1 for hardening. The ultimate survival rate under natural condition was about 80%.Bangladesh J. Sci. Ind. Res. 51(1), 75-80, 2016

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 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.

scholarly journals In vitro regeneration of three varieties of Brassica campestris L. grown in Bangladesh

2020 ◽  
Vol 55 (3) ◽  
pp. 181-188
Author(s):  
B Goswami ◽  
MI Hoque ◽  
S Khan ◽  
RH Sarker
Keyword(s):  
Shoot Regeneration ◽  
Brassica Campestris ◽  
In Vitro Regeneration ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Root Induction ◽  
Regeneration System ◽  
Cotyledonary Leaf ◽  
Best Response

A reproducible in vitro regeneration system was developed for three varieties of Brassica campestris L. namely Agrani, BINA Sarisha-10 and BINA Sarisha-6 using hypocotyl and cotyledonary leaf with petiole as explants. MS medium supplemented with 2.0 mg/l BAP and 0.2 mg/l IAA was found to be the best for the multiple shoot formation for all the three varieties. Among three varieties, BINA Sarisha-6 showed best response in terms of shoot regeneration as well as number of shoots per explant (9.0) using hypocotyls as explants. In case of Agrani and BINA Sarisha-10 highest number of shoot per explants were found 8.2 and 7.0, respectively. Interestingly in vitro regenerated shoots of BINA Sarisha-6 and BINA Sarisha-10 were produced in vitro flowers on shoot regeneration media. Best root induction in BINA Sarisha-6, Agrani and BINA Sarisha-10 was achieved on MS media supplemented with 0.5 mg/l IBA. After proper hardening, the in vitro regenerated plantlets were successfully transplanted into soil. Bangladesh J. Sci. Ind. Res.55(3), 181-188, 2020

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 High Efficiency In vitro Regeneration and Genetic Stability of Corallocarpus epigaeus - An Endangered Medicinal Plant

2020 ◽  
Vol 30 (2) ◽  
pp. 219-229
Author(s):  
Spoorthi Veera ◽  
Pavani Chirumamilla ◽  
Shasthree Taduri
Keyword(s):  
High Efficiency ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Multiple Shoots ◽  
Inter Simple Sequence Repeat ◽  
Multiple Shoot ◽  
Nodal Explants ◽  
Multiple Shoot Induction ◽  
Endangered Medicinal Plant

Indirect regeneration of plantlets from multiple shoot induction of Corallocarpus epigaeus was obtained from leaf and nodal explants on MS with different concentrations of BAP in combination with IAA/IBA or IBA alone. Among all the combinations, BAP and IBA exhibited maximum regeneration. High frequency of multiple shoots (89%) was obtained on BAP (2.0 mg/l) and IBA (1.5 mg/l) in nodal explants. Maximum mean shoot length of 6.8 ± 0.33 cm was obtained in nodal explants cultured on BAP (1.0 mg/l) + IBA (0.5 mg/l), followed by leaf explants with 6.7 ± 0.47 cm on BAP (3.0 mg/l) + IAA (2.5 mg/l). The highest frequency of rooting (88.3%) was obtained on NAA (1.0 mg/l) and IBA (2.0 mg/l) with 21.83 ± 0.57 mean number of roots. The well-rooted healthy plantlets were acclimatized with a survival rate of 80%. Inter simple sequence repeat (ISSR) analysis revealed the genetic similarity of in vitro raised plants with the mother plant. Plant Tissue Cult. & Biotech. 30(2): 219-229, 2020 (December)

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