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 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 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 In vitro regeneration system in brinjal (Solanum melongena L.) for stress tolerant somaclone selection

1970 ◽  
Vol 7 (2) ◽  
pp. 253-258 ◽  
Author(s):  
A Ferdausi ◽  
UK Nath ◽  
BL Das ◽  
MS Alam
Keyword(s):  
Callus Induction ◽  
Biochemical Markers ◽  
Solanum Melongena ◽  
Basal Medium ◽  
Shoot Tip ◽  
Root Induction ◽  
Regeneration System ◽  
Short Period ◽  
Regenerated Plantlets

Brinjal is the second most important vegetable crop after potato in Bangladesh in respect of total areas and third in production. It also plays a vital role in the national economy as a cash crop. An experiment was conducted with two cultivars of brinjal namely Jhumky and Islampuri to observe the callus induction ability of different explants-shoot tip, hypocotyl and midrib in MS basal media supplemented with different concentrations and combinations of growth regulators. The rate of callus induction from shoot tip, hypocotyl and midrib were 82.78%, 74.88% and 78.71%, respectively. Highest rate of callus induction was found in shoot tip. Variety Islampuri showed higher rate of callus induction (80.62%). Among the treatments 2mg/l NAA showed the best performance in callus proliferation. Cytokinin (0.5 mg/l BAP) showed highest percentage of shoot regeneration (57.13%). For root induction, MS basal medium was proved to be better treatment for average number (12-15) of roots. The survival rate of transferred regenerated plantlets after hardening was higher in Jhumky (80%). Regenerated plantlets from callus of both the varieties exhibited 4-9 times higher proline, 2-3 times lower vitamin C and 2-3 times higher iron (Fe) content compared to their seed derived seedlings. This experiment showed that it is possible to develop shoot and fruit borer tolerance brinjal genotypes through somatic embryogenesis that was selected based on biochemical markers within the very short period of time. These findings will be helpful for further selection of the selected variants in field condition in the next phase of the study. Keywords: Somaclonal variants; Biochemical markers; Brinjal DOI: 10.3329/jbau.v7i2.4731 J. Bangladesh Agril. Univ. 7(2): 253-258, 2009

scholarly journals In vitro Micropropagation of Abutilon indicum L. through Leaf Explants

1970 ◽  
Vol 19 (2) ◽  
pp. 177-184 ◽  
Author(s):  
Jyoti Ranjan Rout ◽  
Manorama Mishra ◽  
Ritarani Das ◽  
Santi Lata Sahoo
Keyword(s):  
Callus Induction ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Plantlet Regeneration ◽  
Root Induction ◽  
Ex Vitro ◽  
In Vitro Micropropagation ◽  
Half Strength ◽  
Abutilon Indicum

The present investigation was conducted to develop a protocol for rapid callus induction and plant regeneration from leaf explant of Abutilon indicum L. Callus induction and plantlet regeneration at various frequencies were observed on MS using different concentrations of 2,4-D alone or in combination with BAP and Kn. The highest percentage of callus induction was observed with 2.5 mg/l 2,4-D (90) and with combination of 0.5 mg/l Kn (85). Optimum shoot formation was observed on same medium but supplemented with 2.0 mg/l Kn and 1.0 mg/l NAA (11.2). Rooting experiments with half strength of MS revealed that NAA was more suitable for root induction compared to IBA and IAA. The healthy in vitro rooting plantlets were successfully transferred to the field. The survival of the plantlets under ex vitro condition was 87%. Key words: Abutilon indicum, Callus induction, Leaf explants, Micropropagation D.O.I. 10.3329/ptcb.v19i2.5435 Plant Tissue Cult. & Biotech. 19(2): 177-184, 2009 (December)

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 An Efficient Tissue Culture Regeneration System for Georgia Plume, Elliottia racemosa, a Threatened Georgia Endemic

HortScience ◽  
2008 ◽  
Vol 43 (2) ◽  
pp. 447-453 ◽  
Author(s):  
Seong Min Woo ◽  
Hazel Y. Wetzstein
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Dry Weight ◽  
Medium Composition ◽  
Induction Medium ◽  
Ex Vitro ◽  
Regeneration System

Georgia plume, Elliottia racemosa Muhlenb. ex. Elliott, is an extremely rare small tree or shrub endemic to Georgia, which is being severely affected by habitat loss and lack of sexual recruitment. In vitro plant regeneration of Georgia plume has not been previously reported and may be a method for the conservation and propagation of this threatened species. Studies evaluated the effects of sterilization methods, explant types, medium composition, and light environment on plant regeneration. An efficient plant regeneration system was developed in which adventitious shoot buds were induced using young, expanding leaf explants placed on an induction medium supplemented with 10 μm thidiazuron and 5 μm indole-3-acetic acid with Gamborg's B5 salts. Shoot elongation was promoted on a medium with 25 μm (2-isopentenyl) adenine incorporated into Woody Plant Medium. In vitro rooting studies evaluated continuous and pulse auxin treatments and ex vitro rooting trials after KIBA (indole-3-butric acid, potassium salt) dips. A 5-day pulse treatment on 100 or 150 μm indole-3-butyric acid produced ≈90% rooting of shoots with greater shoot and root dry weight than other pulse times. High rooting frequencies were obtained under in vitro and ex vitro conditions with over 85% survival of plantlets transferred to greenhouse conditions. The culture protocol was found to be effective with material collected from mature specimens in the wild from divergent populations. Tissue culture appears to be a promising approach for the propagation and conservation of this rare and threatened plant.

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 Optimization of IBA for Efficient Survival of Micropropagated Garcinia indica Choisy Shoots Using in-vitro and ex-vitro Rooting Techniques

2019 ◽  
pp. 1-9
Author(s):  
Gauri Mulik ◽  
Shubha Manvi ◽  
Gauri Ingale ◽  
Varsha A. Parasharami
Keyword(s):  
Tissue Culture ◽  
Farmyard Manure ◽  
Ex Vitro Rooting ◽  
In Vitro Rooting ◽  
Root Induction ◽  
Ex Vitro ◽  
Interesting Phenomenon ◽  
National Chemical Laboratory ◽  
Garcinia Indica

Aims: The study focuses on optimization of concentration and time of rooting hormone exposure of IBA (Indole-3-Butyric Acid) for efficient survival of tissue culture raised Garcinia indica Choisy plantlets for in-vitro and ex-vitro rooting techniques. Study Design: The subcultured microshoots of Garcinia indica were subjected to in-vitro and ex-vitro rooting trials by treating them with IBA of varying concentration and time, to standardize these particular parameters required by this auxin to induce rooting. Place and Duration of Study: Rooting trials were carried out in Plant tissue culture-Biochemical Sciences Division of CSIR- National Chemical Laboratory, Pune 411008 between June 2018 and April 2019. Methodology: Regularly subcultured five to six years old shoots from female trees of Garcinia indica were used for the study. Various concentrations of IBA in correlation with time were used for in-vitro and ex-vitro root induction. The rooted plantlets were then transferred to polyhouse for acclimatization and will further be planted in open field locations in June 2019. Results: Induction of rooting was observed within thirty days of treatment with IBA. It was observed that 500ppm of IBA gave 30% rooting for in-vitro rooting trials whereas 2000ppm of IBA induced 80% rooting for shoots given ex-vitro rooting treatments. An interesting phenomenon that was observed for 70% of the shoots which failed to induce rooting by in-vitro treatment was that they survived with 100% rooting success under ex-vitro rooting conditions. The hardened plantlets were successfully acclimatized in the polyhouse with survival rate of 90% and were further transferred to polythene bags with rooting mixtures of sand: farmyard manure: coco peat (1:1:2). These plantlets have been healthy for the last 6 to 9 months and will be transported for field trials in June 2019. Conclusion: Ex-vitro rooting technique was found to be more effective than in-vitro rooting. Thus, by optimizing the rooting hormone parameters, female plants of Garcinia indica can be successfully grown using tissue culture technology and can be propagated in large numbers to increase the female plant number in plantations.

FACTORS AFFECTING THE REGENERATION OF PEPPER (CAPSICUM ANNUUM L.)

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1120G-1120
Author(s):  
J. L. Jacobs ◽  
C. T. Stephens
Keyword(s):  
Growth Hormone ◽  
Silver Nitrate ◽  
Callus Formation ◽  
Leaf Explants ◽  
Regeneration System ◽  
Factors Affecting ◽  
Nitrate Concentrations ◽  
Leaf Differentiation ◽  
Whole Plants

Several growth hormone combinations and silver nitrate concentrations were examined for their effect on regeneration of different pepper genotypes. Primary leaf explants from in vitro seedlings were cultured on a revised Murashige and Skoog medium supplemented with auxin, cytokinin and 1.6% glucose. Combinations of different concentrations of indole-3-acetic acid (IAA), 0-5 mg/l, and 6-benzylaminopurine (BAP), 0-5 mg/l, were tested to determine the most effective medium for shoot primordium formation. Experiments with IAA and BAP did not result in a specific growth hormone combination appropriate for regeneration of all genotypes tested. Of the silver nitrate concentrations tested, 10 mg/l resulted in the best shoot and leaf differentiation and reduced callus formation. Differences in organogenic response of individual genotypes were evaluated on a single regeneration medium. Whole plants were regenerated from 11 of 63 genotypes examined. Based on these experiments, a reproducible regeneration system for pepper was developed with a total of 500 plants regenerated to date.

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