In Vitro Rapid Propagation of Nelumbo nucifera

doi:10.11983/CBB21020

Abstract

Abstract: Using fresh embryos isolated from developing lotus seeds of 18 days after pollination as explants, a rapid propagation system of lotus was established through primary culture, subculture plantlets hardening and transplantation. Results showed that the primary culture medium of MS+0.5 mg·L^-1 6-BA+0.5 mg·L^-1 NAA+30 g·L^-1 sucrose+0.5 g·L^-1 activated carbon+0.8 g·L^-1agar was the optimum for the explant growth. The explant induction rate was up to 85% after cultured for 60 days, Qiuhongyang had the highest amount of stem nodes. In the subculture, the optimal concentration of sucrose in the medium was 80 g·L^-1. When dividing aseptic seedling for subculture, cutting with two stem nodes showed the highest multiplication coefficient. The multiplication coefficient of different varieties was ranged from 4.0 to 6.7, and Qiuhongyang was the highest (6.7). Aseptic seedlings were suggested to be subcultured every 50 days, with high multiplication rates for up to 6 rounds of subcultures. The rooted plantlets were transplanted to pots containing medium of peat:pond soil=1:2 (v/v) during May to July, and the survival rate of transplanted plants was higher than 83.9%. Based on these results, using this in vitro propagation system, it is estimated that 1 465 seedlings be developed from a single lotus seed within one year. This study has established a rapid in vitro propagation system for lotus, which provides the basis for the factory scale production of lotus plantlets.

Key words: lotus, aseptic seedlings, rapid propagation, plant regeneration

Yaqian Xiong, Xianbao Deng, Huihui Zhang, Dong Yang, Heng Sun, Juan Liu, Mei Yang. In Vitro Rapid Propagation of Nelumbo nucifera[J]. Chinese Bulletin of Botany, 2021, 56(5): 605-613.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB21020

https://www.chinbullbotany.com/EN/Y2021/V56/I5/605

Figures/Tables 8

Table 1 The component of the primary culture medium

Types	NAA (mg∙L^-1)	6-BA (mg∙L^-1)	Sucrose (g∙L^-1)	Activated charcoal (g∙L^-1)
I	0.5	0.5	30	0.5
II	0.5	1	30	0.5
III	0.5	0.5	80	0.5
IV	0.5	0.5	30	1

Table 2 The combinations of plant hormones in subculture medium

Types	NAA (mg∙L^-1)	6-BA (mg∙L^-1)
1	0.5	0.5
2	0.3	0.5
3	0.1	0.5
4	0.5	1.5
5	1	0.5
6	0.3	1

Figure 1 Primary culture of Qiuhongyang immature embryo explants from developing seeds of 18 days after pollination (A) Immature embryo explants from developing seeds of 18 days after pollination; (B) Immature embryo cultured one day after inoculation; (C) The rooted plantlets at 10 days after primary culture; (D) The aseptic seedling at 60 days after primary culture. Bars=1 cm

Table 3 Effects of medium components on the growth of Qiuhongyang aseptic seedlings in the primary culture (60 days after inoculation)

Types	The amount of inoculation	Rooting rate (%)	The amount of nodes	The amount of apical bud	The amount of leaves
I	331	100	3.9±0.36 a	0.9±0.03 a	4.3±0.48 a
II	134	98	4.0±0.05 a	0.7±0.04 b	3.8±0.27 a
III	122	95	3.2±0.19 b	0.7±0.01 b	4.3±0.13 a
IV	300	95	3.6±0.33 ab	0.9±0.13 a	3.9±0.22 a

Figure 2 Effects of lotus varieties on the number of stem nodes (A) and the induction rate (B) of lotus aseptic seedlings in the primary culture Different lowercase letters indicate significant differences (P<0.05).

Figure 3 The multiplication coefficient and mortality rate of subculture of aseptic seedlings (A) Effect of dividing methods in Qiuhongyang aseptic seedlings on the multiplication coefficient and mortality rate of subculture (x: Cuttings with two nodes, y: Cuttings with one node and one growing tip, z: Cuttings with one nodes, w: Cuttings with one growing tip); (B) Effects of plant hormone combinations in Qiuhongyang on the multiplication coefficient and mortality rate of subculture, combinations of plant hormones (No.1-6) are in Table 2; (C) Effect of lotus varieties on the multiplication coefficient and mortality rate of subculture. Different lowercase letters indicate significant differences at P<0.05.

Figure 4 Hardening and transplanting of Qiuhongyang aseptic seedlings (A1), (A2) Aseptic seedlings at 50 days of subculture; (B1), (B2) Plantlets at 10 days after transplanting; (C1) Standing leaves started to emerge at 40 days after transplanting; (C2) Rhizomes at 40 days after transplanting; (C3) Rhizome taken from plants grew in ponds. (A1), (A2), (B1), (B2) Bars=1 cm, (C1)-(C3) Bars=10 cm

Table 4 Effects of lotus varieties on the survival rate and the number of leaves developed after transplanting

Lotus varieties	Survival rate (%)		The amount of expanded leaves		The amount of standing leaves
Lotus varieties	10 d	20 d	10 d	20 d	50 d
Qiuhongyang	93.7±3.0 a	83.9±1.6 a	0.4±0.10 b	1.8±0.07 b	0.7±0.04 a
Qiurihonghua	93.3±6.7 a	93.3±6.7 a	0.7±0.07 a	2.7±0.15 a	0.5±0.04 ab
WBG_S1	95.8±7.2 a	93.3±6.3 a	0.5±0.00 b	1.7±0.05 b	0.3±0.06 b
Baihuajianlian	87.8±6.3 a	87.2±18.0 a	0.5±0.11 b	2.5±0.40 a	0.5±0.13 ab

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