scholarly journals In-vitro Callus Induction and Regeneration of Brinjal (Solanum melongena L.) through Cotyledon

Author(s):  
Shreedhar Ganapati Bhat ◽  
G. Arulananthu ◽  
N. Ramesh

Brinjal is one of the most popular, nutritional and vegetable crops in the world. It plays a vital role in the national economy as a cash crop. Tissue culture techniques used for in-vitro plant regeneration through cotyledon explants of eggplant (Solanum melongena L.) with different combinations of plant growth hormones BAP (4.44, 6.66, 8.88, 11.10 and 13.32µM) and IAA (0.57, 1.14 and 1.71µM) used for in-vitro regeneration of brinjal. The cotyledon explants used in this study, the highest callus induction found on BAP 8.88 µM and IAA 1.14 µM. The callus induction occurred after 15days from initiation, shoot induction occurred after 30 days from initiation and shoot elongation was carried out on the same medium, shoot elongation occurred after 45 days from initiation. MS hormone-free medium found best for root regeneration, the elongated shoots were selected and transferred to a test tube containing MS hormone-free rooting medium and the elongated shoots produce roots after 15 days. Then the rooted plantlets were transferred to poly-cup with a pre-sterilized mixture of coco peat for primary hardening under poly-tunnel for 10days. Subsequently, there generated plantlets acclimatized under the greenhouse. Then, hardened plants transferred to the open field for further development. This plant regeneration method can be useful for the production of the disease-free plant.

2012 ◽  
Vol 60 (1) ◽  
pp. 47-55 ◽  
Author(s):  
A. Ali ◽  
T. Yossef ◽  
A. El-Banna

The present study was carried out for developing an efficient in vitro callus induction and plant regeneration system in four different tomato genotypes (Solanum lycopersicum Mill., previous name: Lycopersicon esculentum), Advantage II, Edkawy, Castle Rock and Super Strain B, using hypocotyl and cotyledon explants. The effects of two cytokinins, BA (benzyl adenine) and Kin (kinetin), on callus induction and plant regeneration frequency were investigated when added to MS medium in combination at varying concentrations. All concentrations of the two cytokinins were suitable for callus induction and plant regeneration. The frequency of callus induction and plant regeneration from both cotyledon and hypocotyl explants reached 100% for all tested genotypes. Cotyledons produced a higher average number of shoots per explants than hypocotyls for all the genotypes in the five concentrations of combined cytokinins. The average number of shoots per explant in Super Strain B was found to be the highest (42 and 60 for the hypocotyl and cotyledon explants, respectively). Supplementing MS medium with 1.0 mg L−1 kinetin and 1.0 mg L−1 benzyl adenine was found to be optimum for producing the highest number of shoots per explant from hypocotyls and cotyledons in the tomato genotypes investigated. The proposed medium showed a significant superiority over the reference media.


1970 ◽  
Vol 36 (3) ◽  
pp. 397-406 ◽  
Author(s):  
BP Ray ◽  
L Hassan ◽  
KM Nasiruddin

The effect of different explants and concentrations of BAP and NAA on induction of callus and plant regeneration of brinjal cv. Jhumki were investigated. The treatment combinations were BAP (0. 2.0. 3.0, and 4.0 mg/l) and NAA (0. 0.1, 0.5, and 1.0 mg/l). The rate of callus formation varied in different treatments. The highest amount of callus (48.66%) was produced on MS medium containing 2.0 mg/l BAP and 0.5 mg/l NAA from stem, and 8.2 days required for callus induction. The highest fresh weight of callus was 1.12g from stem and 0.48g from root. The number of shoot regenerated through callus from stem containing 2.0 mg/l BAP and 0.5 mg/l NAA was 3.4 (23.287%) and days required for 38.8 days. All regenerated plantlets survived in normal environment. Keywords: NAA; BAP; regeneration; brinjal. DOI: http://dx.doi.org/10.3329/bjar.v36i3.9268 BJAR 2011; 36(3): 397-406


2019 ◽  
Vol 20 (9) ◽  
pp. 2079 ◽  
Author(s):  
Brenda A. López-Ruiz ◽  
Vasti T. Juárez-González ◽  
Estela Sandoval-Zapotitla ◽  
Tzvetanka D. Dinkova

In vitro plant regeneration addresses basic questions of molecular reprogramming in the absence of embryonic positional cues. The process is highly dependent on the genotype and explant characteristics. However, the regulatory mechanisms operating during organ differentiation from in vitro cultures remain largely unknown. Recently, miRNAs have emerged as key regulators during embryogenic callus induction, plant differentiation, auxin responses and totipotency. Here, we explored how development-related miRNA switches the impact on their target regulation depending on physiological and molecular events taking place during maize Tuxpeño VS-535 in vitro plant regeneration. Three callus types with distinctive regeneration potential were characterized by microscopy and histological preparations. The embryogenic calli (EC) showed higher miRNA levels than non-embryogenic tissues (NEC). An inverse correlation for miR160 and miR166 targets was found during EC callus induction, whereas miR156, miR164 and miR394 displayed similar to their targets RNA accumulation levels. Most miRNA accumulation switches took place early at regenerative spots coincident with shoot apical meristem (SAM) establishment, whereas miR156, miR160 and miR166 increased at further differentiation stages. Our data uncover particular miRNA-mediated regulation operating for maize embryogenic tissues, supporting their regulatory role in early SAM establishment and basipetala growth during the in vitro regeneration process.


2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Shakra Jamil ◽  
Rahil Shahzad ◽  
Ghulam Mohyuddin Talha ◽  
Ghazala Sakhawat ◽  
Sajid-ur-Rahman ◽  
...  

Sugarcane contributes 60–70% of annual sugar production in the world. Somaclonal variation has potential to enhance genetic variation present within a species. Present study was done to optimize an in vitro propagation protocol for sugarcane. The experiments included four varieties, 9 callus induction media, 27 regeneration media, and 9 root induction media under two-factor factorial CRD. Data were recorded on callus induction, embryogenic callus formation, shoot elongation (cm), root induction, and plant regeneration. Statistically significant differences existed between genotypes and treatments for callus induction (%), embryogenic callus formation (%), shoot elongation (cm), root induction, and plant regeneration (%). All parameters showed dependency on genotypes, culture media, and their interaction. Highest callus induction (95%) embryogenic callus formation (95%) was observed in callus induction media 5. Highest plantlet regeneration (98.9%) capacity was observed in regeneration media 11 whereas maximum shoot elongation (12.13 cm) and root induction (8.32) were observed in rooting media 4. G1 showed best response for all traits and vice versa for G4. Hence it was concluded that G1, callus induction media 5, regeneration media 11, and rooting media 4 are the best conditions for in vitro propagation of sugarcane.


HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 572c-572
Author(s):  
Hak-Tae Lim ◽  
Kei-youn Lee ◽  
Yeoung-Sook Yoo ◽  
Duck-Chun Yang

Since in vitro regeneration and transformation systems in hot pepper (Capsicum annuum L.) have not been available, the application of new genetic manipulations has been limited. Here we report an efficient procedure to regenerate whole pepper plants and to generate transgenic plants expressing a foreign gene was established. High frequency of plant regeneration was observed when hypocotyl and cotyledon explants were cultured on MS/B5 medium supplemented with NAA 0.05 mg·L–1 plus zeatin 2.0 mg·L–1, NAA 0.05 mg·L–1 plus zeatin 2.0 mg·L–1, IBA 10.0 mg·L–1 plus BA 1.0 mg·L–1, IAA 0.02 mg·L–1 plus zeatin 3.0 mg·L–1. An addition of AgNO3 5–10 μm to these media improved the regeneration rate by about 10%. For plant transformation, hypocotyl and cotyledon explants of pepper were preconditioned on kanamycin-free shoot induction medium for 48 hours. Then, co-cultivation with Agrobacterium tumeaacience was done on the co-culture medium for 2 days. The explants were then blotted in sterile filter paper and placed on shoot induction and selection medium containing kanamycin sulfate (100 mg·L–1) and carbenicillin (500 mg·L–1). PCR showed that the introduced ADA gene was integrated and stably expressed in the regenerated plants. ADA enzyme activities were checked by spectrophotometric analysis.


Author(s):  
RK Niroula ◽  
BP Sah ◽  
HP Bimb ◽  
S Nayak

This study was under taken to elucidate the effect of genotypes and media compositions on callus induction from mature rice seeds. Three different callus induction media, designated as A (N6 mineral salts + N6 vitamins, 2 mg/l each + myoinositol, 100 mg/l + 2,4-D, 2.5 mg/l + kinetin, 0.5mg/l + AgNO3, 10 mg/l + maltose, 50 gm/l); B (MS organic salts + N6 mineral salts + NAA, 4 mg/l + kinetin, 2 mg/l + AgNO3, 5mg/l and sucrose, 60 gm/l); and C (B media without AgNO3), and six rice genotypes viz. Jumlimarshi, Tilki, Jethobudo, Manshara, Masuli and Pahenle were evaluated. The modified N6 medium supplemented with 2, 4-D, 2.5 mg/l and AgNO3, 10 mg/l exhibited better performance in callus induction. Among genotypes, callus induction frequency was higher (100%) in Masuli, Tilki and Jumlimarshi regardless of media tested. The positive effect of AgNO3 was only observed in medium A for quality callus induction and subsequent plant regeneration. The genotype Tilki performed better regarding plant regeneration (27.77%). Therefore, it is suggested that application of medium A is advantageous to accomplish overall efficiency of callus induction and plant regeneration from seeds of various rice genotypes. Key words: 2, 4-D; AgNO3; dehulled rice; in vitro; regeneration J. Inst. Agric. Anim. Sci. 26:21-26 (2005)


HortScience ◽  
2008 ◽  
Vol 43 (1) ◽  
pp. 215-219 ◽  
Author(s):  
Yeh-Jin Ahn ◽  
Grace Qianhong Chen

An efficient plant regeneration protocol using cotyledon explants was established for castor (Ricinus communis L.), an important oilseed crop. Mature seed-derived cotyledon explants produced adventitious shoots when placed on Murashige and Skoog (MS) medium containing thidiazuron (TDZ). The rate of shoot regeneration was maximal (≈25 shoots per explant) when explants were cultured on shoot induction medium supplemented with 5 μm TDZ and preincubated in the dark for the first 7 days before transferring to the day/night cycle (16/8 h). Only the proximal ends of cotyledon explants produced adventitious shoots, although green calli were observed in cotyledon veins. After 4 weeks in culture, explants with well-developed shoot buds were transferred to MS medium without plant growth regulators for the shoot elongation and development. At ≈4 months after culture initiation, shoots (2 cm in length) were transferred to root induction medium (MS medium supplemented with 5 μm indole-3-butyric acid) where they developed roots in 4 to 6 weeks. Plantlets were transferred to soil and acclimatized to greenhouse conditions. Histological analysis showed the adventitious induction of the shoots originated from the cortical and epidermal cell layers of the cotyledon explants.


2015 ◽  
Vol 25 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Kee Hwa Bae ◽  
Eui Soo Yoon

Lychnis cognate Maxim and Lychnis fulgens Fish. Ex Spreng are two valued ornamental plants in Korea. Soaking of seeds in GA3 solution remarkably promoted germination up to 60%, but the control (0 mg/l) was not effective (> 5%). To select an adequate temperature for seed germination, seeds, previously soaked in a 1000 mg/l GA3 for 24 hrs, were incubated at 15, 20, 25, and 30°C. Seed germination of over 20% was obtained at 15, 20, and 25°C, but only 10% at 30°C. These results indicate that the seeds of L. cognate and L. fulgens are in a such dormant state that they hardly germinate even by dormancy breaker (GA3) and low (15 ? 25°C) temperature treatment. The highest callus induction was observed in the leaf explants of the seedlings on MS containing specific concentrations of 3.0 mg/l BA and 1.0 mg/l NAA. The adventitious shoot was formed < 90% of calli on 1/2 WPM medium. The height of in vitro propagated plantlet was no different media used for regeneration. This in vitro propagation protocol should be useful for conservation of endangered and ornamental plant.Plant Tissue Cult. & Biotech. 25(1): 1-12, 2015 (June)


Biologia ◽  
2010 ◽  
Vol 65 (4) ◽  
Author(s):  
Sara Rostampour ◽  
Haleh Sohi ◽  
Ali Dehestani

AbstractPersian poppy (Papaver bracteatum Lindl.) is an important commercial source of medicinal opiates and related compounds. In this research, calli were induced from seeds, roots, cotyledons and hypocotyls of P. bracteatum at a high efficiency. The optimized callus induction media consisted of the Murashige and Skoog (MS) basic media supplemented with 1.0 mg/L 2, 4-dichlorophenoxyacetic acid (2,4-D), 0.1 mg/L kinetin and 15 mg/L ascorbic acid. The concentrations of 2,4-D and ascorbic acid were found critical to callus induction and proliferation. Subsequent subcultures resulted in excellent callus proliferation. Ascorbic acid at concentration 15 mg/L increased the callus proliferation significantly. Maximum callus growth was achieved when the explants were incubated at 25°C. MS salts at full strength were found inhibitory for callus induction, while ľ MS salts were found to favor callus induction. Shoot regeneration of calli in vitro was achieved on ľ MS medium containing 0.5 mg/L benzylamine purine and 1.0 mg/L naphthalene acetic acid. Analysis of alkaloid extracts from Persian poppy tissues by high-performance liquid chromatography showed that thebaine accumulated in the tissues of plants. The thebaine alkaloid profile of the Persian poppy is a well-defined model to evaluate the potential for metabolic engineering of thebaine production in P. bracteatum.


1970 ◽  
Vol 35 (2) ◽  
pp. 331-341 ◽  
Author(s):  
MA Sayem ◽  
M Maniruzzaman ◽  
SS Siddique ◽  
M Al-Amin

The experiment was conducted to investigate the performance of three different genotypes (BARI Sarisha-6, BARI Sarisha-8, and BARI Sarisha-11) in two different media viz., MS and B5 with different concentrations of phytohormone (2, 4-D) for callus induction from uninucleate stage anthers of Brassica and subsequent plant regeneration in MS media with different concentrations of phytohormone (BAP and NAA). Among the genotypes, BARI Sarisha-8 showed the best performance for all the parameters of callus induction. The performance of BARI Sarisha-6 was poor compared to others. Maximum rate of callus induction (%) was observed in MS + 0.5 mg/L 2, 4-D followed by B5 + 0.5 mg/L 2,4-D. The media combination MS + 1.0 mg/L BAP 0.3 mg/L 2,4-D showed the best performance for maintenance of calli. Significant variations were observed among the genotypes and media composition for shoot regeneration. Among the genotypes, BARI Sarisha-8 showed the best performance for shoot regeneration followed by BARJ Sarisha-l1. The genotype BARI Sarisha-8 produced higher percent of shoots/calli and required minimum days for shoot initiation. Higher percent calli without shoot were produced by the genotype BARI Sarisha-6. The media combination MS + 2.0 mg/L BAP + 0.5 mg/L NAA showed the best performance for shoot regeneration and required maximum days for shoot initiation. Keywords: Regeneration; BARI Sarisha-6; BARI Sarisha-8; BARI Sarisha-11; anther culture; phytohormone  DOI: 10.3329/bjar.v35i2.5896Bangladesh J. Agril. Res. 35(2) : 331-341, June 2010


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