Abstract
Determination of genetic stability of in vitro-grown plantlets is needed for safe and large-scale production of mature trees. In this study, genetic variation of long-term micropropagated mature pistachio developed through direct shoot bud regeneration using apical buds (protocol A) and in vitro-derived leaves (protocol B) was assessed via DNA-based molecular markers. Randomly amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR), and amplified fragment length polymorphism (AFLP) were employed, and the obtained PIC values from RAPD (0.226), ISSR (0.220), and AFLP (0.241) showed that micropropagation of pistachio for different periods of time resulted in “reasonable polymorphism” among donor plant and its 18 clones. Mantel’s test showed a consistence polymorphism level between marker systems based on similarity matrices. In conclusion, this is the first study on occurrence of genetic variability in long-term micropropagated mature pistachio plantlets. The obtained results clearly indicated that different marker approaches used in this study are reliable for assessing tissue culture-induced variations in long-term cultured pistachio plantlets.
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Abbreviations
- AFLP:
-
Amplified fragment length polymorphism
- BA:
-
6-Benzyladenine
- IAA:
-
Indole acetic acid
- IBA:
-
Indole-3-butyric acid
- EMR:
-
Effective multiplex ratio
- Ib av :
-
Average band informativeness
- ISSR:
-
Inter-simple sequence repeat
- MI:
-
Marker index
- PCA:
-
Principal component analysis
- PC:
-
Principal coordinate
- PIC:
-
Polymorphic information content
- RAPD:
-
Randomly amplified polymorphic DNA
- RFLP:
-
Restriction fragment length polymorphism
- R p :
-
Resolving power
- SSR:
-
Simple sequence repeat
- TC:
-
Tissue culture
- UPGMA:
-
Unweighted pair-group method with arithmetic
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Acknowledgments
The authors are grateful to MSc. Emrah Kirdok for helping to statistical analysis. This research was funded by a grant no. KBAG 209 T030 from The Scientific and Technological Research Council of Turkey (TUBITAK).
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Supply. Fig. 1
Diagram of plant regeneration protocol A and B for mature P. vera L. (JPG 105 kb)
Supply. Fig. 2
All PCR programmes used in this study. (DOCX 218 kb)
Supply. Fig. 3
Examples of RAPD patterns assessed with K-01 primer; lanes D: Donor plant, G: Greenhouse material, 1-10: 5-years micropropagated plants, 20-26: 7-years micropropagated plants, “M: marker (some polymorphic bands were shown with arrowheads)”. (DOCX 281 kb)
Supply. Fig. 4
Examples of ISSR patterns assessed with ISSR-12 primer; lanes D: Donor plant, G: Greenhouse material, 1-10: 5-year micropropagated plants, 20-26: 7-years micropropagated plants, “M: marker (some polymorphic bands were shown with arrowheads)”. (DOCX 374 kb)
Supply. Fig. 5
Examples of AFLP patterns assessed with E-AAC and M-CTA primer combinations; lanes D: Donor plant, G: Greenhouse material, 1-10: 5-year micropropagated plants, 20-26: 7-years micropropagated plants, “M: marker (some polymorphic bands were shown with arrowheads)”. (DOCX 460 kb)
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Akdemir, H., Suzerer, V., Tilkat, E. et al. Detection of Variation in Long-Term Micropropagated Mature Pistachio via DNA-Based Molecular Markers. Appl Biochem Biotechnol 180, 1301–1312 (2016). https://doi.org/10.1007/s12010-016-2168-7
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DOI: https://doi.org/10.1007/s12010-016-2168-7