scholarly journals Efficient regeneration protocol for callus and shoot induction from recalcitrant Phaseolus vulgaris L. explants under optimum growth conditions

2021 ◽  
Vol 8 (4) ◽  
Author(s):  
Alfred Besra ◽  
Jolly Basak
Keyword(s):  
Phaseolus Vulgaris ◽  
Callus Induction ◽  
Growth Conditions ◽  
Root Tip ◽  
Induction Medium ◽  
Root Induction ◽  
Shoot Induction ◽  
Phaseolus Vulgaris L ◽  
Transgenic Lines ◽  
Shoot Induction Medium

Callus is the most significant morphogenic response obtained in plant tissue culture studies. It can be used for micropropagation or to create transgenic lines. Phaseolus vulgaris L. (common bean) is one of the economically important crops with a great nutritional value. However, very little effort has been made to regenerate callus from P. vulgaris explants. Six explants were used namely root tip, leaves, plumule, radicle, cotyledon and embryo to develop a callus from P. vulgaris. The minimum days for callus induction was 10 days in plumule, radicle and embryo explants, while the maximum was 15 days in cotyledon explants with the callus induction percentage of 75%. The largest callus was found to be 2.77 gm in weight and 2.5 cm in diameter in MS medium. Medium with different concentrations of plant growth regulators (PGRs) showed different growth pattern in callus induction. Culture medium with 1.50 mg/l of BAP, 0.50 mg/l of 2, 4-D and 0.10 mg/l of NAA showed the best result in callus induction. Higher concentration of BAP (2.00 mg/l), along with 0.25 mg/l of 2, 4-D was ideal for shoot regeneration and maturation. Shoot induction medium along with 2.00 mg/l of NAA concentrations were found to be best for rooting. It was found that plumule and radicle favor callus induction, however, they are also potent for shoot and root induction. Knowledge gained in this study will be useful in developing a standard protocol for plant regeneration from P. vulgaris explants and will also be useful in creating transgenic line of P. vulgaris.

A micropropagation protocol for Melaleuca alternifolia (tea tree)

1996 ◽  
Vol 36 (6) ◽  
pp. 755 ◽  
Author(s):  
SE List ◽  
PH Brown ◽  
CS Low ◽  
KB Walsh
Keyword(s):  
Multiple Shoots ◽  
Induction Treatment ◽  
Induction Medium ◽  
Root Induction ◽  
Shoot Induction ◽  
Melaleuca Alternifolia ◽  
Tea Tree ◽  
Shoot Induction Medium ◽  
Basal Salts ◽  
Stem Segments

Micropropagation of Melaleuca alternifolia (tea tree) was achieved using stem segments from mature plants. A comparison of the source of explant material, and the form of cytolunin and mineral nitrogen on the induction of multiple shoots per node is reported. Further, the effect of shoot induction treatment, the age of plant used for explants and the value of the presence of charcoal, sucrose and light on subsequent root induction in plantlets is considered. A culture procedure involving nodes sourced from 1.5 to 3 cm from the shoot tip, cultured into a shoot induction medium of Murashige and Skoog (MS) basal salts supplemented with 4.5 �mol 6-benzylaminopurine/L and 60 mmol sucrose/L for 12 weeks, and then subcultured for root induction on medium containing MS basal salts, 60 mmol sucrose/L and 0.1 or 1 �mol indole-acetic acid/L for 8 weeks, is recommended.

Development of an Agrobacterium-mediated Transformation System for `Beurre Bosc' Pear

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 461d-461
Author(s):  
Richard L. Bell ◽  
Ralph Scorza ◽  
Chinnathambi Srinivasan
Keyword(s):  
Shoot Proliferation ◽  
Induction Medium ◽  
Shoot Induction ◽  
Transformation System ◽  
Gus Histochemical Assay ◽  
Young Leaves ◽  
Leaf Tissues ◽  
Shoot Induction Medium ◽  
Basal Salts

An efficient regeneration/transformation system was developed for `Beurre Bosc' pear. Young leaves were harvested from in vitro shoots proliferated on a medium containing MS basal salts and 5 BAP, 0.5 μM IBA, and 0.6M3. Shoot regeneration was optimized using a modification of the medium of Chevreau and Leblay (1993). Explants were cultured on shoot induction medium contained 10 μM TDZ and 1 μM IBA for 4 weeks in the dark, and then transfered to a similar, but auxinless, regeneration medium until shoots developed, usually after an additional 4 to 8 weeks. Leaf tissues were transformed by co-cultivation for 3 days with Agrobacterium tumefaciens EHA101 carrying a pGA482 plasmid containing NPTII, GUS, and rolC genes, followed by cultivation on SIM containing 300 mg/L timentin. Putative transgenic plants were selected on shoot induction medium containing 80mg/L kanamycin, and multiplied on shoot proliferation medium. Four clones were confirmed as transgenic using the GUS histochemical assay and Southern blots for the NPTII and rolC genes. Plants of each clone have been rooted and successfully transfered to the greenhouse for further analysis of gene expression.

scholarly journals Silicon Promotes Adventitious Shoot Regeneration and Enhances Salinity Tolerance ofAjuga multifloraBunge by Altering Activity of Antioxidant Enzyme

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Iyyakkannu Sivanesan ◽  
Byoung Ryong Jeong
Keyword(s):  
Salinity Tolerance ◽  
Shoot Regeneration ◽  
Microscopic Analysis ◽  
Adventitious Shoot ◽  
Induction Medium ◽  
Shoot Induction ◽  
Electron Microscopic ◽  
Sod Activity ◽  
Scanning Electron Microscopic Analysis ◽  
Shoot Induction Medium

We investigated the effect of Si concentration on shoot regeneration and salinity tolerance ofAjuga multiflora. Addition of Si to the shoot induction medium significantly increased the frequency of shoot induction. The average number of shoots regenerated per explant decreased on the medium containing NaCl alone, while there was less decrease when the shoot induction medium was supplemented with both NaCl and Si. The shoot induction percentage increased linearly with increasing concentration of Si in the NaCl containing medium. Addition of Si to the shoot induction medium significantly increased SOD, POD, APX, and CAT activity in regenerated shoot buds as compared with the control. The inclusion of Si to the NaCl containing medium significantly increased the SOD activity in leaves and roots, while it decreased POD, APX, and CAT activity in both organs. Scanning electron microscopic analysis showed that there are no distinct differences in the structure of stomata between the control and Si-treated plants. However, NaCl treatment significantly affected the structure and number of stomata as compared to the control. Wavelength dispersive X-ray analysis confirmed the high Si deposition in trichomes of plants grown in the Si containing medium but not in plants grown in the medium without Si.

scholarly journals Establishment of a Rapid and Efficient Micropropagation System for Succulent Plant Haworthia turgida Haw.

HortScience ◽  
2017 ◽  
Vol 52 (9) ◽  
pp. 1278-1282 ◽  
Author(s):  
Boling Liu ◽  
Hongzhou Fang ◽  
Chaorong Meng ◽  
Ming Chen ◽  
Qingdong Chai ◽  
...  
Keyword(s):  
Callus Induction ◽  
Adventitious Shoots ◽  
Germplasm Conservation ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Root Induction ◽  
Ms Medium ◽  
Succulent Plant

In the present study, the effect of plant growth regulators (PGRs) on callus regeneration, adventitious shoot differentiation, and root formation of Haworthia turgida Haw. was investigated. The greatest callus induction percentage (95.6%) was achieved with leaf explants inoculated on Murashige and Skoog (MS) medium with 1.0 mg·L−1 6-benzyladenine (BA) and 0.1 mg·L−1 1-naphthaleneacetic acid (NAA), and this callus induction medium supplemented with 2.5 mg·L−1 thidiazuron (TDZ) was optimal for callus proliferation. The maximum number of shoots (25.7) was obtained when the callus was cultured on MS medium supplemented with 1.0 mg·L−1 BA and 0.2 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D). The highest number of roots per shoot (6.2) and highest rooting frequency (82.0%) were obtained when adventitious shoots were inoculated on MS medium with 0.05 mg·L−1 NAA. Regenerated plantlets were transferred to a mixture of vermiculite and soil and acclimated in a greenhouse. The survival rate of the transplanted plantlets was about 91.6%. The rate of ex vitro rooting was 83.3%, indicating that this technique is effective for root induction in H. turgida. This study has established a rapid and efficient micropropagation system that can be beneficial for commercial cultivation and germplasm conservation of H. turgida.

scholarly journals Efficient Plant Regeneration from Cotyledon and Midrib Derived Callus in Eggplant (Solanum melongena L.)

1970 ◽  
Vol 14 ◽  
pp. 31-38 ◽  
Author(s):  
M Rahman ◽  
M Asaduzzaman ◽  
N Nahar ◽  
MA Bari
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Key Words ◽  
Somatic Embryo ◽  
Callus Induction ◽  
Somatic Embryos ◽  
Solanum Melongena ◽  
Induction Medium ◽  
Root Induction ◽  
Ms Medium

Somatic embryos were obtained from cotyledon and midrib explants of Solanum melongena L., cultivar Loda. For callus induction, medium was supplemented with different concentrations of auxin singly or in combination with BAP. The best callusing 83-85% was obtained from both of the explants cultured on MS medium containing 2.0 mgl-1NAA + 0.05 mgl-1BAP. Somatic embryogenesis and shoot regeneration was achieved after transferring the calli to MS medium supplemented with BAP, GA3, NAA and Zeatin. Cotyledon derived calli showed better performance (87%) for regeneration than that of midrib (82%) when sub cultured on MS medium having 2.0 mgl-1 Zeatin + 1.0 mgl-1 BAP. For root induction, MS + 3.0 mgl-1 IBA was proved to be better treatment for average number (14-15) and mean length (12 cm) of roots than those of other treatments. Key words: Eggplant; cotyledon; midrib; callus induction; somatic embryo J. bio-sci. 14: 1-9, 2006

scholarly journals A New Method for Rapid In Vitro Propagation of Apple and Pear

HortScience ◽  
2001 ◽  
Vol 36 (6) ◽  
pp. 1102-1106 ◽  
Author(s):  
V.R. Bommineni ◽  
H. Mathews ◽  
S.B. Samuel ◽  
M. Kramer ◽  
D.R. Wagner
Keyword(s):  
Clonal Propagation ◽  
Shoot Multiplication ◽  
Induction Medium ◽  
Shoot Induction ◽  
Clonal Multiplication ◽  
Propagation Methods ◽  
Shoot Induction Medium ◽  
Tools And Techniques ◽  
Stem Slices

Improved in vitro clonal propagation methods are valuable tools for nurseries and growers, and are essential for manipulation and improvement of tree fruit germplasm using the tools and techniques of biotechnology. We have developed a rapid shoot multiplication procedure for clonal propagation of apple, Malus ×domestica cv. Gale Gala and pear, Pyrus communis L. cv. Bartlett. Rapid clonal multiplication was achieved after the following series of steps: pre-conditioning of micropropagated shoots, sectioning pre-treated stems into thin slices, placing slices onto shoot induction medium and incubating directly under cool-white fluorescent lights or after a brief dark incubation. Multiple induction of shoots recovered from stem slice explants within three weeks of culture. A maximum of 37% of cultured apple stem slices, and 97% of pear stem slices, showed induction of shoots. More shoots were recovered on phytagel solidified shoot induction medium than on agar. Cultured stem slices of both apple and pear showed maximum recovery of shoots from shoot induction medium supplemented with thidiazuron (TDZ) compared to medium supplemented with BAP and kinetin. Under ideal conditions, pear stems generated four times the shoots as the same quantity or length of apple shoots. Micropropagated shoots were rooted and transferred to the greenhouse and field nursery for further evaluation. Chemical names used: N-phenyl-N′-1,2,3-thidiazol-5-ylurea (thidiazuron or TDZ); 6-benzylaminopurine (BAP).

scholarly journals GENOTYPIC DIFFERENCES IN MORPHOGENIC POTENTIAL OF CULTURED LEAF EXPLANTS OF LYCOPERSICON HIRSUTUM

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1064G-1064
Author(s):  
John R. Stommel
Keyword(s):  
Callus Induction ◽  
Wild Species ◽  
Indoleacetic Acid ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Induction Medium ◽  
Genotypic Differences ◽  
Lycopersicon Hirsutum ◽  
Shoot Induction ◽  
Wild Tomato

Cultured leaf explants obtained from 36 accessions of the wild tomato Lycopersicon hirsutum were evaluated for morphogenic capacity in response to 3 cytokinins [zeatin, benzylamino purine (BA) and kinetin] in combination with indoleacetic acid (IAA). Morphogenic responses within this wild species were accession-dependent, Cotyledon tissue, in comparison to true leaf explants, were superior for callus and shoot formation. Optimal callus induction medium varied with accession, but most often contained 13.3 μM BA plus 1.7 μM IAA. Media containing 4.6 or 9.2 μM zeatin plus 0.1 μM iaa were optimal shoot induction media. Explants of L. hirsutum f. typicum accessions 126445, 127826, 128644, and 390663 and L. hirsutum f. glabratum accessions 365904, 365905, and 365906 exhibited the highest levels of shoot formation.

scholarly journals GENOTYPIC DIFFERENCES IN MORPHOGENIC POTENTIAL OF CULTURED LEAF EXPLANTS OF LYCOPERSICON HIRSUTUM

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1064g-1064
Author(s):  
John R. Stommel
Keyword(s):  
Callus Induction ◽  
Wild Species ◽  
Indoleacetic Acid ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Induction Medium ◽  
Genotypic Differences ◽  
Lycopersicon Hirsutum ◽  
Shoot Induction ◽  
Wild Tomato

Cultured leaf explants obtained from 36 accessions of the wild tomato Lycopersicon hirsutum were evaluated for morphogenic capacity in response to 3 cytokinins [zeatin, benzylamino purine (BA) and kinetin] in combination with indoleacetic acid (IAA). Morphogenic responses within this wild species were accession-dependent, Cotyledon tissue, in comparison to true leaf explants, were superior for callus and shoot formation. Optimal callus induction medium varied with accession, but most often contained 13.3 μM BA plus 1.7 μM IAA. Media containing 4.6 or 9.2 μM zeatin plus 0.1 μM iaa were optimal shoot induction media. Explants of L. hirsutum f. typicum accessions 126445, 127826, 128644, and 390663 and L. hirsutum f. glabratum accessions 365904, 365905, and 365906 exhibited the highest levels of shoot formation.

scholarly journals Agrobacterium-mediated Transformation of Three Elite Hybrid Aspens

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 535B-535
Author(s):  
M.J. Bosela ◽  
J.P. Schnurr ◽  
Z.-M. Cheng ◽  
W.A. Sargent
Keyword(s):  
Shoot Formation ◽  
Histochemical Staining ◽  
Kanamycin Resistance ◽  
Induction Medium ◽  
Shoot Induction ◽  
Transformation Protocol ◽  
High Frequencies ◽  
Transformation Experiment ◽  
Shoot Induction Medium ◽  
Hybrid Clones

Three elite hybrid aspen, Populus grandidentata × P. canescens, P. tremuloides × P. tremula, and P. tremuloides × P. davidiana, have been transformed with Agrobacterium tumefaciens strains LBA4404 and EHA105 carrying kanamycin resistance and GUS genes. The leaves of micropropagated shoots were co-cultivated with Agrobacterium for 65 to 72 hr and then transferred to callus-induction medium with 80–120 mg/L kanamycin in the dark. After 2 weeks, the leaves were transferred to shoot-induction medium under 18-hr photoperiod. Regenerated shoots were verified for transformation by histochemical staining and PCR. Transformed shoots rooted and were transplanted to soil. The three hybrid clones differed widely in their medium requirements for regeneration and in their competence for transformation. The leaves of P. grandidentata × P. canescens callused vigorously on a wide variety of media. In a typical transformation experiment, 30% to 60% of infected leaves produced putatively transformed calli (up to 10 calli per leaf). The origin of these calli and the frequency of shoot formation depended on the Agrobacterium strains. The calli from EHA105-infected leaves produced shoots within six weeks of co-cultivation and at high frequencies (70% to 90%). However, the calli from LBA4404-infected leaves produced shoots more slowly and at much lower frequencies (5% to 10%). Delaying selection for 2 weeks was found to lower the transformation frequency. Putatively transformed calli were obtained from P. tremuloides × P. tremula, and P. tremuloides × P. davidiana hybrids at frequencies of only 2% to 3%. The calli regenerated from P. tremuloides × P. davidiana leaves were very small, but they continued to grow upon being transferred to shoot-induction media and have started to produce shoots. The calli from leaves of P. tremuloides × P. tremula were much larger and they produced shoots more quickly. This transformation protocol is currently being used to introduce rooting genes into these hybrids to improve their rooting from hardwood cuttings.

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