Plant Regeneration from in Vitro-Selected Salt Tolerant Callus Cultures of Solanum Tuberosum L (original) (raw)
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In vitro selection of salt tolerant cell lines in Solanum tuberosum L
Biologia Plantarum, 2007
Cell lines able to grow on media containing 50, 100, 150 or 200 mM NaCl were established from potato callus cultures by direct recurrent selection or gradual selection. In callus subjected to direct selection only small clusters of cells survived on medium with 150 or 200 mM NaCl, whereas on 100 mM small cell portions appear necrotic. When cell lines were obtained by successive subcultures on media with increased concentrations of NaCl, salt-tolerant calli were more compact and developed a greenish colour free from necrotic areas. The response of calli lines grown on media with NaCl was compared to control line. The NaCl-tolerant calli showed a decrease in relative growth rate and water content, with higher reductions in the 150 mM tolerant callus. Lipid peroxidation was increased in 50 mM and 100 mM NaCl-tolerant calli, while in 150 mM tolerant callus remained similar to 100 mM values. There was a significant increase in ascorbic acid content in 100 mM and 150 mM NaCl-tolerant calli as compared to the 50 mM, that was two-fold the value found in the control. Also, the contents of soluble and insoluble proteins increased in salt-tolerant lines. SDS-PAGE of soluble proteins showed the synthesis of specific polypeptides in the presence of NaCl in culture medium and the synthesis of a new polypeptide.
New Zealand Journal of Crop and Horticultural Science, 2001
A comparative growth analysis was made between salt‐tolerant plants obtained by recurrent selection methods in vitro and Solanum tuberosum L. ‘Kennebec’ original mother plants (wild type). They were grown at three different salt concentrations (0.5, 25, and 100 mMNaCl) by hy‐droponic culture. Fourteen days after the saline treatment was applied, no differences in growth were observed between clones. However, 14 days later (Day 28), the comparative analysis between clones revealed intra‐especific differences. The salt‐tolerant line (clone 150) exhibited greater tolerance to the highest salt doses. Plants of this clone had greater biomass than wild type (clone T) at all salt concentrations tested. The relative growth rate (RGR) and components, as well as the relative accumulation rate of water (RARwater) and K+ and Na+ content, should explain, at least in part, the salt‐tolerance of clone 150.
In Vitro regeneration in two potato (Solanum tuberosum L.) varieties Cardinal and Heera
Two potato varieties (Cardinal and Heera) were used in ln Vitro regeneration experiment. Leaf and internodes were used as explants to observe the callus induction and plantlet regeneration ability of two potato varieties The explants were cultured on MS medium supplemented with different concentrations and combinations of growth regulators. The percentage of callus induction ranged from 20 to 100 %. Highest percentage of callus induction (100%) was observed in MS medium supplemented with 5.0 mg/L NAA and 2.0 mg L-1 BAP. Cardinal showed higher callusing than Heera. Internode explants of cv. Cardinal required minimum time (9.67 days) for callus initiation in MS medium supplemented with 5.0 mg/L NAA and 4.0 mg L-1 BAP. Cardinal showed the best performance for regeneration. internode of Cardinal had the highest number of regenerated plants per vial (3.0) in MS medium supplemented with 3.0 mg L-1 NAA and 4.0 mg L-1 BAP, where as leaf of Heera gave minimum number (0.67) of plantlets per vial in the medium with 5.0 mg L-1 NAA and 2.0 mg L-1 BAP
The identification of the best culture medium and the most suitable tomato variety was perform ed by In vitro regeneration. In the present study, attem pts have been made to develop an efficient protocol for regeneration of V1 (ROMA VF ), V2 (Baromasi Hybrid Tom ato: JHOLOK) and V3 (T omato Hybrid F1 JAGU R) varieties with their tolerance against abiotic stress (salt) to obtain stress tolerant tom ato. The cotyledon induced in MS medium which was supplemented with T1 (MS+1mg/l BA+0.5mg/l NAA), T2 (MS + 1.5mg/l BA + 1mg/l NAA) and T3 (2mg/l BA + 1.5mg/l N AA) from seeds of those varieties. Maximum percentage (75%) of cotyledon induction was observed in T1 for V1, V2 and T3 for V3 respectively. Cotyledon derived from different concentrations of BA and NAA were cultured on MS m edium supplem ented with T3 (1. 5 mg/l NAA, 2 mg/l BA) and TL (2.0 mg/l Kinetin) for plantlet regeneration. It was observed that MS m edia supplemented with T3 + TL produced lowest percentage of shoot (20%), callus (10%) and root (0%) in case of V2 and highest percentage of shoot (80%), callus(60%) and root (20%) in case of V3. Callus were transferred to the regener ation medium supplemented with NaCl (0 to 75mM) in order to examine their responses to salinity, the above three varieties, showed a significant decline in the callus growth. Present studies have shown that Hybrid F1 JAGUR variety is more responsive in regeneration and salt tolerant than others.
A stable salt-tolerant potato cell line, able to grow on media containing 60-450 mM NaCl (i.e. low to high salinity) was selected. Callus grown on 120 or 150 mM NaCl showed higher fresh weights than the rest of the treatments. Replacing NaCl by KCl or Na 2 SO 4 showed that reductions in fresh weight were mainly due to the presence of Na + ions. When PEG 6000 was added to the medium instead of salt, the salt tolerant cell lines were unable to overcome the PEG-induced water stress. Whole plants, regenerated from salt tolerant callus, exhibited salt stress tolerance as evidenced by their higher fresh and dry weights when watered with 90 mM NaCl, and they also produced more tubers per plant under salt stress. Salt-tolerant plants differed phenotypically from control plants both in terms of leaf shape, tuber flesh and skin colour, which was reddish. In addition, DNA fingerprinting by RAPDs, with 70 different primers, confirmed that the salt tolerant regenerants also differed genotypically from the control, salt sensitive Kennebec potato plants from which they had been selected.
Plant Cell Tissue and Organ Culture, 1998
A stable salt-tolerant potato cell line, able to grow on media containing 60–450 mM NaCl (i.e. low to high salinity) was selected. Callus grown on 120 or 150 mM NaCl showed higher fresh weights than the rest of the treatments. Replacing NaCl by KCl or Na2SO4 showed that reductions in fresh weight were mainly due to the presence of Na+ ions. When PEG 6000 was added to the medium instead of salt, the salt tolerant cell lines were unable to overcome the PEG-induced water stress. Whole plants, regenerated from salt tolerant callus, exhibited salt stress tolerance as evidenced by their higher fresh and dry weights when watered with 90 mM NaCl, and they also produced more tubers per plant under salt stress. Salt-tolerant plants differed phenotypically from control plants both in terms of leaf shape, tuber flesh and skin colour, which was reddish. In addition, DNA fingerprinting by RAPDs, with 70 different primers, confirmed that the salt tolerant regenerants also differed genotypically from the control, salt sensitive Kennebec potato plants from which they had been selected.
Biocatalysis and Agricultural Biotechnology, 2020
Callus induction and its subsequent regeneration is an important issue to study the genetic variability in breeding and biotechnological program. Under this study, various physiological characteristics of calli were evaluated based on ten (10) indigenous and six (6) exotic potato genotypes. Correlation matrix, mean rank, and standard deviation of rank related to the physiological states were also studied. According to the mean rank and standard deviation of rank, potato genotypes were classified into three distinct groups such as good (rank sum ranges 4.65-10.17), fair (10.53-13.96) and poor (14.21-16.40). In this study, Surjamukhi, Granola, Sheelbilati, Arun and Sindurkouta exhibited good callusing while Courage, Diamant, Jamalu, Sadaguti, Patnai and Cardinal showed fair index. Lalpakri, Asterix, Chollisha, Dohazari and Ausha displayed low callusing index. Among the tested genotypes ANOVA, DMRT and correlation coefficient of all in vitro callus inductions characteristics were found highly significant at p < 0.01 levels. Calli of the studied genotypes were cultured in MS medium in addition with BAP (5.0 mg/L) + IAA (2.0 mg/L) + GA 3 (1.0 mg/L) for regeneration. Maximum shoot regeneration was recorded in Sadaguti (40%) with a highest number of shoot (21.3). Meanwhile, lowest regeneration rate was obtained in Chollisha (19.99%) and the shoot number was 11.0. The standardized protocols and the message of the present findings on callus induction and their regeneration efficiency may be helpful to evaluate the rest indigenous and exotic cultivars of potatoes and its further improvement through biotechnological approaches in Bangladesh.
Effect of Different Medium on Callus Induction and Regeneration in Potato Cultivars
The present study was undertaken to develop an effective protocol for optimum callus induction and plant regeneration in 4 potato varieties; Arnova, Burren, Provento and Riviera. Different combinations of hormones (2mg/l BA +2.5mg/l NAA; 2mg/l BA+ 2mg/l 2,4-D ; 2mg/l 2,4-D) with control treatment (hormone free) were tested for callus induction. After first, second and third subculture, callus were transferred to regeneration media that contained different combinations of hormones (2.5mg/l BA+5mg/l GA3; 3mg/l BA+0.5mg/l GA3+ 0.03mg/l NAA; 0.22mg/l TDZ+ 0.49mg/l NAA; 5 mg/l TDZ) with control treatment (without hormone). Data of % callus induction, number of days required for callus induction, callus morphology, callus fresh weight, number of days required for regeneration, % regeneration, number of shoots/callus clump, shoot length, number of nodes/shoot and number of leaves/shoot were taken. Stem segments of one clone from Provento, Burren and Riviera were planted on tuberization medium to study the effect of varieties on microtuber induction potential. Results showed that there were significant differences among varieties; Burren and Riviera had the highest % callus induction, fresh weight and number of days for callus induction. A medium containing 2 mg/l BA + 2.0 mg/l 2,4-D and 2 mg/l 2,4-D alone gave good response and a good callus proliferation. The results concerning with regeneration revealed that when callus transferred to regeneration media after first subculture, excellent regeneration was observed in a medium with 3mg/l BA+ 0.5mg/l GA3 + 0.03 mg/l NAA in Burren , Provento and Riviera, while in Arnova variety shoot regenerated only on media containing 0.22mg/l TDZ+ 0.49mg/l NAA. Shoot formation completely failed when callus after second and third subculture transferred to regeneration media. For microtubrizaition, differences were detected among the cultivars in all characteristics studied except tuber number.
African Journal of …, 2010
tuberosum L.) cultivar Diamant. The tuber segments were used as explants and cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of -naphthalene acetic acid (NAA), 2,4-Dichlorophenoxy acetic acid (2,4-D), benzyl adenine (BA) and thidiazeron (TDZ) alone and 2,4-D in combinations with BA for callus induction. The best degree for callus formation (6.0) was obtained on MS medium supplemented with 2,4-D alone at 3.0 mg/l or 2,4-D in combination with BA both at 2.0 mg/l. MS media supplemented with different levels of BA and TDZ were employed for shoot regeneration. MS medium containing 5.0 mg/l TDZ was the best for days to shoot initiation, the highest percentage of callus with shoot (81%) and highest number of shoot per callus (3.4). Callus derived shoots were rooted most effectively in half-strength MS medium containing 0.5 mg/l IBA. The success of plant tissue culture for in vitro culture of potato was encouraged by acclimatization of the plantlets in the greenhouse conditions. Regenerated plants were morphologically uniform with normal leaf shape and growth pattern.
Callus Formation and Organogenesis of Potato (Solanum tuberosum L.) Cultivar Almera
The Journal of Phytology, 2010
A procedure for plant regeneration from callus culture of potato, Solanum tuberosum L. is described. Calli were induced from 1.0 cm 2 tuber segment of potato cultivar Almera on Murashige and Skoog's medium (MS) supplemented with different levels (1.0-5.0 mg/l) of 2, 4-dichlorophenoxy acetic acid (2, 4-D). The highest degree of callus formation (3.0) and hundred percent (100%) of explants produced nodular calli on MS medium within 7-12 days when supplemented with 2.0-5.0 mg/l of 2, 4-D. Calli were differentiated into shoot-primordia when subcultured on MS medium supplemented with 1.5-5.0 mg/l of thidiazuron (TDZ) and 2.0-5.0 mg/l of benzyladenine (BA). The best result for number of shoot per callus (3.3 ± 0.3) and longest shoot (0.8 ± 0.1) were obtained by using TDZ at 5.0 mg/l. Callus derived shoots were rooted most effectively in full-strength MS medium containing 1.0 mg L-1 IBA. The success of plant tissue culture for in vitro culture of potato was encouraged by acclimatization of the plantlets in the greenhouse conditions. Regenerated plants were morphologically uniform with normal leaf shape and growth pattern.