Recurrent somatic embryogenesis and development of somatic embryos in Akebia trifoliata (Thunb.) Koidz (Lardizabalaceae)
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A simple and effective protocol was established for recurrent somatic embryogenesis and plant regeneration in Akebia trifoliata (Thunb.) Koidz. Somatic embryos directly formed from the root zone of the immature zygotic embryos cultured on MS medium devoid of plant growth regulator (PGR). The induction frequency of immature zygotic embryos was 52.5%, and the mean number of somatic embryos reached up to 19.5. Secondary somatic embryos arose on the primary embryos at a high frequency (95.8%), and this process maintained in a recurrent way on PGR-free MS medium. The highest mean number of somatic embryos also appeared in the primary embryos which can reach up to 44.7. While both of the embryogenic potential and mean number of embryos per explant displayed a gradual diminution with subculturing. The addition of 0.5 mg l−1 6-benzyladenine in the 1/2 MS regeneration medium can significantly improve the frequency of somatic embryos convert to plantlets. The microscopic analysis revealed that the development process of somatic embryos via the globular, heart, torpedo and cotyledonal stages in A. trifoliata, the histological analysis showed that somatic embryos directly initiated from root epidermal cells and there are no vascular connections between the somatic embryos and maternal tissue. Regenerated plantlets acclimated successfully to greenhouse conditions. Approximately 65% plantlets survived and displayed no morphological characteristics differences with seed-derived plants. The simple and effective protocols established in this study will promote large-scale clonal propagation and genetic improvement of A. trifoliata.
Somatic embryogenesis has never been reported in Akebia trifoliata (Thunb.) Koidz. The study showed the origin and development process of somatic embryos in Akebia trifoliata (Thunb.) Koidz.
KeywordsAkebia trifoliata Somatic embryogenesis Somatic embryos Development Hormone-free medium
This work was supported by the Plant Germplasm Innovation Program, Science and Technology Service Network Initiative, Chinese Academy of Sciences (ZSZC-007).
HH conceived and designed the experiments, SZ carried out the study and wrote a draft, HH and XY revised the manuscript, CZ, DL, ZW contributed to the writing of the manuscript. All authors read and approved the manuscript final version.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
- Abohatem MA, Bakil Y, Baaziz M (2017) Plant regeneration from somatic embryogenic suspension cultures of date palm. Date Palm Biotechnol Protoc 1637:203–214Google Scholar
- Burbulis N, Kupriene R (2005) Induction of somatic embryos on in vitro cultured zygotic embryos of spring Brassica napus. Acta Univ Latviensis 691:137–143Google Scholar
- Halperin W (1995) In vitro embryogenesis: some historical issues and unresolved problems. In: Thorpe TA (ed) In vitro embryogenesis in plants. Springer, Dordrecht, pp 1–16Google Scholar
- Klimaszewska K, Overton C, Stewart D, Rutledge RG (2011) Initiation of somatic embryos and regeneration of plants from primordial shoots of 10-year-old somatic white spruce and expression profiles of 11 genes followed during the tissue culture process. Planta 233:635–647PubMedCrossRefPubMedCentralGoogle Scholar
- Liu LP, Qian ZX (2002) Determination of nutritional components in fruit of Akebia trifoliate Koidz. J Southeast Guizhou Natl Teach Coll 20:39–41Google Scholar
- Lu D, Wei W, Zhou W, McGuigan LD, Ji F-y, Li X, Xing Y, Zhang Q, Fang K-f, Cao Q-q, Qin L (2017) Establishment of a somatic embryo regeneration system and expression analysis of somatic embryogenesis-related genes in Chinese chestnut (Castanea mollissima Blume). Plant Cell, Tissue Organ Cult 130:601–616CrossRefGoogle Scholar
- Merkle SA (1995) Strategies for dealing with limitations of somatic embryogenesis in hardwood trees. Plant Tissue Cult Biotechnol 1:112–121Google Scholar
- Normah M, Rohani E, Mohamed-Hussein Z (2013) Somatic embryogenesis in higher plants. Malays Appl Biol 42:1–12Google Scholar
- Norusis MJ (1998) SPSS/PC advanced statistics. SPSS Inc, ChicagoGoogle Scholar
- Park YS, Lelu-Walter MA, Harvengt L, Trontin JF, MacEacheron I, Klimaszewska K, Bonga JM (2006) Initiation of somatic embryogenesis in Pinus banksiana, P. strobus, P. pinaster, and P. sylvestris at three laboratories in Canada and France. Plant Cell, Tissue Organ Cult 86:87–101CrossRefGoogle Scholar
- Pavlović S, Vinterhalter B, Zdravković-Korać S, Vinterhalter D, Zdravković J, Cvikić D, Mitić N (2012) Recurrent somatic embryogenesis and plant regeneration from immature zygotic embryos of cabbage (Brassica oleracea var. capitata) and cauliflower (Brassica oleracea var. botrytis). Plant Cell, Tissue Organ Cult 113:397–406CrossRefGoogle Scholar
- Shen GL, Shao AJ, Huang LQ, Lin SF (2007) Studies on callus culture of Akebia trifoliata. China J Chin Mater Media 32:899–901Google Scholar
- Wang DZ, Li F, Yan J, Zhong HM (2004) Study and application of nutritional components of wild plant Var australis (Diels) Rehd. Amino Acids Biot Resour 26:16–17Google Scholar
- Wu LL, Ke BF, Gong C, Ma XL, Li JA (2015) Tissue culture and rapid propagation of Akebia trifoliate var. australis. Plant Physiol J 51:903–908Google Scholar
- Yeung EC (1995) Structural and developmental patterns in somatic embryogenesis. In: Thorpe TA (ed) In vitro embryogenesis in plants. Current plant science and biotechnology in agriculture, vol 20. Springer, DordrechtGoogle Scholar