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. 2022 Aug 12:13:914652.
doi: 10.3389/fpls.2022.914652. eCollection 2022.

Efficient regeneration of mature castanopsis hystrix from in vitro stem explants

Heng Zhang  1   2 , Mengqing Guo  1   2 , Qiaona Wu  1   2 , Mengqiu Zhao  1   2 , Ruiping Li  1   2 , Xiaomei Deng  1   2 , Ruchun Xi  1   2
Affiliations

Efficient regeneration of mature castanopsis hystrix from in vitro stem explants

Heng Zhang et al. Front Plant Sci. .

Abstract

Castanopsis hystrix is one of the main timber trees grown in China. However, severe shortage of natural seeds and the difficulty of explant regeneration has limited seedling supply. As such, there is a need for research on asexual multiplication of C. hystrix. This study established a rapid propagation technology system for C. hystrix genotypes, including explant treatment, proliferation, and rooting. HZ (a modified MS medium) supplemented with 4.4 μM BA and 0.5 μM IBA was found to be the optimal medium for shoot sprouting. The maximum proliferation coefficient and the number of effective shoots was obtained on HZ medium supplemented with 2.6 μM BA and 1.0 μM IBA, were 3.00 and 5.63, respectively. A rooting rate of 83.33% was achieved using half-strength HZ medium supplemented with 3.2 μM NAA. Adding vitamin C (80 mg⋅l-1) for 7 days in a dark environment reduced the browning rate, while increasing the proliferation rate. Additionally, through cytological observation, we established how and where adventitious roots occur. The survival rate of transplanted plantlets was > 90%. This is the first report of an in vitro regeneration technique that uses stem segments of mature C. hystrix as explants.

Keywords: Castanopsis hystrix; adventitious roots; browning; disinfection; in vitro regeneration; mature tree.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Effects of different types and concentrations of anti-browning agents on shoot proliferation and browning. Group a (left) was cultured directly under light. Group b (right) was transferred to light culture after a 7-day darkness treatment. (A) Proliferation coefficient; (B) browning rate. Different uppercase letters in the same column indicate a significant difference, as per Duncan’s multiple range test (p ≤ 0.05, n = 3 indicates three replicates).
FIGURE 2
FIGURE 2
Micropropagation of mature Castanopsis hystrix trees using axillary node explants. (A) Mother plant; (B) the miniature grafting nursery; (C) shoot sprouting; (D,E) shoot proliferation; (F) rooting; (G) plantlet acclimatization for 30 days; (H) plantlet acclimatization for 90 days.
FIGURE 3
FIGURE 3
Histology observation on differentiating roots formation of Castanopsis hystrix plantlets. Ri, pith; Rp, root primordium; Ar, adventitious root; Ca, callus. (a) Rooted on day 4; (b) the cambium cells were active on day 5; (c) callus appeared; (d) root primordium appeared on day 7; (e,f) root primordium differentiated; (g) adventitious root broken through the cortex on day 11; (h) adventitious root broken through callus on days 12–13; (i) the adventitious root vascular connected to the stem vascular.
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