Technology for Rapid Propagation in Vitro of Ancient Four-season Rhododendron Tree in Longyuan of Pingnan

doi:10.3969/j.issn.1000-2561.2020.04.017

Abstract

Abstract:

Rapid propagation in vitro was explored using the apical buds of an ancient Four-season Rhododendron tree, more than 400 years, from Longyuan Village, Tangkou Town, Pingnan County as the explants to find the effects of sterilizing methods and time of explants on the development of sterile cultures, effects of kinds and ratios of plant growth regulators added in the media on the proliferation of shoots and rooting of rootless shoots. The optimum sterilization conditions were the explants treated with 75% of ethanol for 30 seconds and then dipped with 0.1% of HgCl₂for 8 minutes. The optimum sampling period for explants was about middle to late May. After four times of bottle transfer success rate of re-sterilizing reached 71% through rescuing of the contaminated explants in the development of sterile cultures by treating with 75% of ethanol for 20 seconds and then dipping with 0.1% of HgCl₂for 5 minutes. The optimum components of the medium in the primary culture was WPM medium added 3.0 mg/L of zeatin (ZT), 0.1 mg/L of NAA and 0.5 mg/L of GA₃, on which the average number of induced valid buds was 3.45. In the proliferation culture, the clustered shoots were induced from apical and stem segments with axillary buds of sterile plant-lets inoculated on Anderson medium added 0.5 mg/L of TDZ and 0.1 mg/L of NAA, multiplication coefficient of stem segments with axillary buds was higher than that of apical buds, reached 13.23. In the rooting culture of rootless shoots, the optimum medium was WPM medium added 0.1 mg/L of ZT and 0.5 mg/L of NAA, on which the rooting rate reached 92.6%. The results would provide theoretical and technical supports for conservation, exploitation and utilization of the ancient Four-season Rhododendron tree.

Key words: Pingnan Four-season Rhododendron, ancient tree, rapid propagation in vitro

CLC Number:

Q813.1

HU Jihong, CHEN Guixin, YANG Huiting, KANG Jianban, GAN Daikui. Technology for Rapid Propagation in Vitro of Ancient Four-season Rhododendron Tree in Longyuan of Pingnan[J]. Chinese Journal of Tropical Crops, 2020, 41(4): 755-763.

Figures/Tables 11

References 26

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编号 No.	酒精消毒时间 Sterilizing time of ethanol time/s	HgCl₂ 消毒时间Sterilizing time of HgCl₂/min	平均污染率 Average contamination rate/%	平均死亡率 Average mortality rate/%	平均成功率 Average success rate/%
1	30	5	68.3^a	14.9^d	16.8^d
2	30	6	65.2^a	18.8^c	16.0^d
3	30	7	58.4^b	15.8^d	25.8^c
4	30	8	33.5^c	21.9^b	44.6^a
5	30	9	31.3^c	29.9^a	38.8^b

编号 No.	酒精消毒时间 Sterilizing time of ethanol time/s	HgCl₂ 消毒时间Sterilizing time of HgCl₂/min	平均污染率 Average contamination rate/%	平均死亡率 Average mortality rate/%	平均成功率 Average success rate/%
1	30	5	68.3^a	14.9^d	16.8^d
2	30	6	65.2^a	18.8^c	16.0^d
3	30	7	58.4^b	15.8^d	25.8^c
4	30	8	33.5^c	21.9^b	44.6^a
5	30	9	31.3^c	29.9^a	38.8^b

编号 No.	取材时间 Sampling period time/month	消毒时间 Sterilizing time/min	接种数 Number of inoculated buds	平均污染率 Average contamination rate/%	平均死亡率 Average mortality rate/%	平均成功率 Average success rate/%
1	10	8	75	33.5^b	21.9^b	44.6^b
2	3	8	75	25.8^c	48.7^a	25.5^c
3	5	8	75	22.5^d	10.8^c	66.7^a
4	6	8	75	56.3^a	25.5^b	18.2^d

编号 No.	取材时间 Sampling period time/month	消毒时间 Sterilizing time/min	接种数 Number of inoculated buds	平均污染率 Average contamination rate/%	平均死亡率 Average mortality rate/%	平均成功率 Average success rate/%
1	10	8	75	33.5^b	21.9^b	44.6^b
2	3	8	75	25.8^c	48.7^a	25.5^c
3	5	8	75	22.5^d	10.8^c	66.7^a
4	6	8	75	56.3^a	25.5^b	18.2^d

编号 No.	酒精消毒时间 Sterilizing time of ethanol time/s	HgCl₂消毒时间Sterilizing time of HgCl₂/min	次氯酸钠消毒时间 Sterilizing time of sodium hypochlorite /min	接种数 Number of inoculated buds	平均污染率 Average contamination rate/%	平均死亡率 Average mortality rate/%	平均成功率 Average success rate/%
1	30	-	-	45	79.5^a	20.5^c	0^c
2	-	-	15	45	15.2^c	84.8^a	0^c
3	20	-	5	45	60.0^b	40.0^b	0^c
4	20	5	-	45	21.4^c	7.6^e	71.0^a
5	30	5	-	45	19.9^c	15.5^d	64.6^b