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In vitro selection for drought tolerance in black pepper (piper nigrum L.)
The study entitled “In vitro selection for drought tolerance in black pepper” was carried out in the Department of Plant Biotechnology, College of Agriculture, Vellayani during 2014 to 2016.The objective was to develop drought tolerance in black pepper (var. Panniyur-4) by in vitro selection using p...
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| Formato: | Ph.D Thesis |
| Publicado em: |
Vellayani
Department of Plant Biotechnology, College of Agriculture
2017
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| Assuntos: |
| LEADER | 03789nam a22001697a 4500 | ||
|---|---|---|---|
| 082 | |a 660.6 |b LAK/IN | ||
| 100 | |a Lakshmi Krishna | ||
| 245 | |a In vitro selection for drought tolerance in black pepper (piper nigrum L.) | ||
| 260 | |a Vellayani |b Department of Plant Biotechnology, College of Agriculture |c 2017 | ||
| 300 | |a 81p | ||
| 502 | |a MSc | ||
| 520 | 3 | |a The study entitled “In vitro selection for drought tolerance in black pepper” was carried out in the Department of Plant Biotechnology, College of Agriculture, Vellayani during 2014 to 2016.The objective was to develop drought tolerance in black pepper (var. Panniyur-4) by in vitro selection using polyethylene glycol (PEG) and to characterize the tolerance. Callus was induced from different explants of black pepper namely leaf, nodes and shoot tip. Murashige and Skoog’s (MS) medium containing different combinations and concentrations of plant growth regulators (PGR) were used for callus induction. Percentage of callus induction varied among the type of explants used. In the present study leaf tissue was found to be most responsive to callus induction than other explants. Among the fourteen treatments, 1.0 mg L-1 of 6- benzyladenine (BA) recorded the highest callus induction of 75 % from leaves in a period of 45 days. The same treatment produced 50 % callus induction from nodes and 41.66 % from shoot tips. In vitro selection for drought tolerance was done by growing the calli in MS medium containing 1.0 mg L-1 of BA and different concentration of polyethylene glycol ie., 5 g L-1 to 100 g L-1 for 30 days. It was observed that increasing concentration of PEG resulted in reduction of growth and browning of the calli. The calli were able to survive in PEG up to a maximum concentration of 10 g L-1 (83.3 %). None of them survived beyond that concentration of PEG. The PEG tolerant calli were multiplied and maintained in MS supplemented with 1.0 mg L-1 of BA containing the respective concentration of PEG. PEG tolerant callus were subjected to regeneration in MS medium supplemented with different concentrations and combinations of PGRs, but they failed to show regeneration. Biochemical analysis of PEG tolerant calli showed significant difference in proline, total free amino acids, soluble protein and ascorbic acid content. Calli survived in 10 g L-1 of PEG showed the highest amount of proline content (0.87 ug g-1) than the one survived in 5 g L-1 of PEG. The amino acid (2.02 mg g-1), soluble protein (1.71 mg g-1) and ascorbic acid (9.23 mg 100g-1) content were significantly high in calli survived at 10 gL-1 of PEG. The increase in SOD content in PEG tolerant calli was not significant. The RNA isolated from PEG tolerant calli showed a maximum yield of 1600 ng/ul and purity value of 2.3. Amplification of cDNA using Actin gene confirmed the quality of cDNA. Differential expression p5cs (Pyrroline-5- carboxylate synthase) gene was observed in the PEG tolerant calli compared to control. RT-PCR using degenerate primers designed for p5csgene amplified a band of size approximately 120 bp and its expression was high in calli survived in 10 gL-1 of PEG. In the present study, the PEG tolerant calli developed showed high levels of proline, free amino acids, total soluble protein, SOD activity and ascorbic acid content, indicating the development of water stress tolerance. Expression of p5cs gene was also increased in PEG tolerant callus. Hence, this in vitro selection system can be used for the selection of drought tolerance in black pepper. | |
| 650 | |a Plant Biotechnology | ||
| 700 | |a Soni K B (Guide) | ||
| 942 | |2 ddc |c TH | ||
| 999 | |c 157949 |d 157949 | ||
| 952 | |0 0 |1 0 |4 0 |6 660_600000000000000_LAKIN |7 1 |8 REF |9 157739 |a KAUCLV |b KAUCLV |c THESES |d 2017-12-27 |o 660.6 LAK/IN |p 174102 |r 2017-12-27 |w 2017-12-27 |y TH | ||