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. 2023 Oct 9;12(19):3507.
doi: 10.3390/plants12193507.

Shoot Organogenesis and Regeneration from Leaf Seedlings of Diospyros oleifera Cheng

Yang Liu  1   2   3   4   5 , Naifu Zhou  6 , Chengrui Luo  1   4   5 , Qi Zhang  3 , Peng Sun  3 , Jianmin Fu  3 , Shuzhan Li  1   4   5 , Ze Li  1   2   4   5
Affiliations

Shoot Organogenesis and Regeneration from Leaf Seedlings of Diospyros oleifera Cheng

Yang Liu et al. Plants (Basel). .

Abstract

Persimmons (Diospyros) are economically important trees that are widely cultivated for wood production in China, and Diospyros oleifera Cheng is the main persimmon grafting stock. However, an efficient tissue culture system has not been perfected for D. oleifera due to the limits of proliferation and rooting cultures. Therefore, this study examined the effects of different plant growth regulators and concentrations on the primary culture of young embryos, induction of leaf callus, differentiation of adventitious shoots, and rooting culture of D. oleifera. The optimal formula for young embryo germination was 1/2 Murashige and Skoog (MS) medium containing 0.5 mg/L gibberellic acid (GA3); after 25 days, the sprouting rate of the young embryos was 67.3%. The best medium for leaf callus induction was 1/2MS medium containing 2.0 mg/L of 6-benzylaminopurine (6-BA) and 0.5 mg/L of naphthaleneacetic acid (NAA), and the callus induction rate was 88.9%. Then, the callus was transferred to 1/2MS medium containing 2.0 mg/L of zeatin (ZT), 0.5 mg/L of NAA, and 2.0 mg/L of thidiazuron (TDZ) to induce adventitious shoots; after 25 days, 5.4 buds were produced per explant, and the induction rate of the adventitious shoots was 88.3%. The adventitious shoots were transferred to 1/2MS medium containing 2.0 mg/L of ZT, 2.0 mg/L of 6-(γ,γ-dimethylallylamino)purine (2iP), and 0.1 mg/L of indole acetic acid (IAA) for the proliferation culture, for which the multiplication coefficient approached 7.5. After multiplication, the adventitious shoots were inoculated into 1/2MS medium containing 1.0 mg/L of indole butyric acid (IBA), 0.5 mg/L of NAA, and 1.0 mg/L of kinetin (KT); the rooting rate was 60.2%, and the average number of roots was 6.9.

Keywords: Callus induction; adventitious shoots; disinfection; rooting; tissue culture.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Germination of seed embryos of Diospyros oleifera Cheng. (A) Seed embryo germination after 10 days. (B) Seedling growth at 25 days. (C) Secondary culture at 30 days.
Figure 2
Figure 2
Effect of adding different concentrations of sucrose to 1/2MS + 2.0 mg/L 6-BA + 0.5 mg/L NAA medium on leaf callus induction. The same letters in rows are not significantly different at p ≤ 0.05.
Figure 3
Figure 3
Leaf callus induction of Diospyros oleifera Cheng. (A) Leaf discs inoculated into culture medium. (B) Callus formed at the incision after 10 days of leaf culture. (C) Leaf discs were inoculated in culture medium for 20 days. (D) Complete callus tissue formed after 25 days. (E) Adventitious shoots after 15 days of callus culture. (F) Adventitious shoots after growth for 25 days.
Figure 4
Figure 4
Adventitious shoots induced on the leaf callus. (A) Callus-generated adventitious shoots at 15 days. (B) After 25 days, the indefinite buds grew and the leaves unfolded; adventitious shoots grown for (C) 30, (D) 40, and (E,F) 50 days.
Figure 5
Figure 5
Adventitious shoot proliferation culture of Diospyros oleifera Cheng. (A–D) First-generation proliferation culture; (E,F) second-generation proliferation culture. (A,E) Single adventitious shoots were planted in the culture medium and cultured for 10 (B,F), 20 (C,G), and 30 (D,H) days.
Figure 6
Figure 6
Growth of D. oleifera rooting culture under different plant growth regulators for 40 days in (A,E) 1/2MS + 1.0 mg/L IBA + 0.5 mg/L NAA + 1.0 mg/L KT; (B,F) 1/2MS + 1.0 mg/L IBA + 1.0 mg/L KT; (C,H) 1/2MS + 2.0 mg/L IBA; and (D,G) 1/2MS + 1.0 mg/L IBA + 0.5 mg/L NAA + 1.0 mg/L KT.
Figure 7
Figure 7
Rooting culture of D. oleifera in 1/2MS + 1.0 mg/L IBA + 0.5 mg/L NAA + 1.0 mg/L KT medium. The same letters in rows are not significantly different at p ≤ 0.05.
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