麻栎种子和胚方脱水敏感性及其低温贮藏技术

doi:10.11707/j.1001-7488.LYKX20240144

Abstract

Abstract:

Objective: This study aims to investigate the desiccation sensitivity as well as the critical moisture content of the seed and embryonic axis of Quercus acutissim and explore low-temperature preservation techniques for Q. acutissima seeds and ultra-low temperature preservation techniques for the embryonic axis, in order to provide technical guidance for the preservation of recalcitrant seeds such as Q. acutissima. Method: Wild Q. acutissim seeds were used as experimental materials, the silica gel weight reduction method was used to rapidly dehydrate the seeds to the target moisture content, and then the germination indexes of seeds and embryonic axes with different moisture contents were determined. The differential scanning calorimetry (DSC) was employed to determine the thermodynamic parameters of embryonic axes and cotyledons with different moisture contents, and the critical moisture content of crystallization in the embryonic axis was evaluated according to the relationship between moisture content and corresponding enthalpy. The low-temperature sealed storage method was used to preserve seeds with different moisture contents, and the vitrification method was used for cryopreservation of the embryonic axis. Result: 1) When the initial moisture content of the seeds and embryonic axes decreased from 34.90% and 44.14% to 10.00%, their germination percentage decreased from 93.00% and 90.00% to 5.00% and 52.00%, respectively. 2) The DSC results of Q. acutissim embryonic axis and cotyledon with different moisture contents showed that with the decrease of moisture content of embryonic axis and cotyledon, the crystallization initiation temperature, peak temperature and enthalpy value showed a regular downward trend, which were significantly positively correlated with moisture content. The average enthalpy value of the embryonic axis was higher than that of the cotyledon, and the free moisture content of the embryonic axis was higher than that of the cotyledon. It was preliminarily determined that the theoretical critical moisture content of crystallization in this batch of the embryonic axes was 11.72%. 3) The effects of dehydration on the physiological and biochemical characteristics of Q. acutissima seeds showed that as dehydration progressed, the activities of superoxide dismutase (SOD) and peroxidase (POD), as well as the contents of proline (PRO) and malondialdehyde (MDA) in seeds initially increased and then decreased. 4) Q. acutissim seeds with initial moisture content stored at 4 ℃ for 21 days showed no change in the germination vitality, and the seeds with severe dehydration (10%–20%) were still able to tolerate a certain degree of low temperature damage. The results of cryopreservation of embryonic axis with different moisture contents showed that moisture content had a significant impact on the results of cryopreservation. When pretreated with PVS2 for 30 min before cryopreservation, the embryonic axis with 10.00% and 15.00% moisture content had a 5.00% and 3.00% survival rate, respectively, and the thermodynamic characteristics of the embryonic axis with different moisture content changed. The embryonic axis with 15.00% moisture content regained free moisture. Conclusion: The germination rate of Q. acutissim seeds and embryonic axes decreases with the decrease of moisture content, which is a typical characteristics of recalcitrant seeds with the dehydration sensitivity. Dehydration has a significant effect on the physiological and biochemical characteristics of the seeds. The theoretical critical moisture content of crystallization in Q. acutissim embryonic axis is 11.72%. Cryopreservation of Q. acutissimembryonic axis is possible, but the method still needs adjustment to increase the recovery of germination rate, and then is applied to the long-term preservation of the recalcitrant seed.

Key words: Quercus acutissim, desiccation sensitivity, cryopreservation, recalcitrant seeds, differential scanning calorimetry (DSC).

CLC Number:

Mingjia Zhang,Boqiang Tong,Kai Qu,Yang Xian,Chengcheng Cui,Yongzheng Wang,Yicun Zang,Biao Han. Desiccation Sensitivity and Low-Temperature Preservation Techniques of Quercus acutissim Seeds and Embryonic Axes[J]. Scientia Silvae Sinicae, 2025, 61(4): 129-139.

Figures/Tables 12

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References 0

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含水量 Moisture content (%)	实际含水量范围 Actual moisture content range (%)	萌发率 Germination percentage (%)	萌发指数 Germination index	萌发时间 Germination time/d	根长 Length of root/mm	发霉数量 Mildew number
34.90	—	93.00±0.04c	4.36±0.04c	7.49±1.24a	6.59±0.43a	1.66±0.15a
30.00	29.14~30.89	82.00±0.02c	4.12±1.32c	7.81±0.80a	7.02±0.85a	1.66±0.15 a
25.00	24.69~26.05	85.00±0.09c	2.24±0.14b	12.05±0.32ab	6.98±1.63a	1.00±0.00a
20.00	18.32~21.35	42.00±0.02b	1.94±0.06ab	10.34±0.11a	4.32±0.66b	4.33±0.33c
15.00	13.98~15.87	19.00±0.10a	1.03±0.33a	15.92±0.32b	5.18±0.16ab	3.00±0.00b
10.00	8.33~12.50	5.00±0.01a	0.33±0.04a	15.45±1.79b	2.69±0.45b	4.33±0.33c

种子部位 Seed site	含水量 Moisture content (%)	冷却结晶阶段Cooling crystallisation			加热熔融阶段Warming melt
种子部位 Seed site	含水量 Moisture content (%)	起始温度 Onset temperature/℃	峰值温度 Peak temperature/℃	热焓 Enthalpy/(J·g^?1)	起始温度 Onset temperature/℃	峰值温度 Peak temperature/℃	热焓 Enthalpy/(J·g^?1)
胚轴 Embryonic axes	10.00	无结晶峰No peak
	15.00	?34.01±5.84a	?43.50±4.50a	81.60±13.6a	?80.71±67.97a	?10.07±4.30a	48.23±24.01a
	20.00	?32.40±2.51a	?38.70±5.90a	85.50±3.50a	?19.08±0.35a	?9.75±1.05a	49.00±1.00a
	25.00	?31.92±4.02b	?37.52±0.48a	101.37±6.93a	?17.10±0.39a	?8.14±0.44ab	60.11±7.88a
	30.00	?26.29±0.94b	?28.65±3.76ab	142.69±0.02b	?12.62±3.57a	?5.04±2.21ab	75.35±2.34a
	55.33	?18.01±1.73c	?16.30±2.72b	358.14±13.41c	?5.80±0.14a	?0.67±0.11b	143.40±6.28b
子叶 Cotyledon	10.00	无结晶峰No peak
	15.00	无结晶峰No peak
	20.00	?32.19±3.48a	?29.30±1.92a	92.30±5.84a	?15.16±1.31a	?7.13±0.55a	40.05±4.51a
	25.00	?26.70±2.20ab	?23.05±1.05ab	114.11±2.11a	?10.99±0.48b	?4.33±0.22b	42.31±6.68a
	30.00	?21.47±3.02ab	?21.31±2.79ab	129.11±12.89a	?10.06±0.44bc	?4.32±0.21b	42.01±6.38a
	44.25	?18.00±2.21b	?18.62±2.09b	220.06±81.64a	?6.69±1.18c	?2.19±1.01b	80.52±11.95b

含水量 Moisture content (%)	胚轴热焓 Embryonic axes enthalpy/(J·g^?1)			子叶热焓 Cotyledon enthalpy/(J·g^?1)			平均热焓 Average enthalpy/(J·g^?1)
含水量 Moisture content (%)	冷却结晶 Cooling crystallisation	加热熔融 Warming melt	平均热焓 Average enthalpy	冷却结晶 Cooling crystallisation	加热熔融 Warming melt	平均热焓 Average enthalpy	平均热焓 Average enthalpy/(J·g^?1)
10.00	—	—	—	—	—	—	—
15.00	81.60	48.23	65.42	—	—	—	—
20.00	85.50	49.00	71.75	92.30	40.05	71.18	88.90
25.00	101.37	60.11	80.91	114.11	42.31	78.21	107.74
30.00	142.69	75.35	109.02	129.11	42.01	85.56	135.90
34.94	358.14	143.40	250.77	220.06	80.52	150.29	289.10

温度 Temperature/℃	萌发率Germination rate (%)
温度 Temperature/℃	初始含水量Initial moisture content	30.00%	25.00%	20.00%	15.00%	10.00%
4	93.33±4.40a	68.00±2.77a	58.00±10.13b	55.00±10.40b	18.00±4.40b	15.00±2.88c
–20	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00
–40	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00
–80	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00

胚方大小 Embryonic axis size/g	胚方体积 Embryonic axis square volume/cm³	萌发率 Germination percentage（%）	染菌率 Contamination rate（%）
2.50~3.00	3.00~3.20	88.00±4.4a	48.33±8.33a
1.50~2.00	1.20~1.50	85.00±0.00a	41.66±7.26a
0.05~1.00	0.30~0.40	92.00±1.66a	3.44±1.66b

Desiccation Sensitivity and Low-Temperature Preservation Techniques of Quercus acutissim Seeds and Embryonic Axes

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含水量 Moisture content (%)	初始萌发率 Initial germination rate (%)	超低温保存成活率Cryopreservation survival rate (%)
含水量 Moisture content (%)	初始萌发率 Initial germination rate (%)	15 min	30 min	45 min	60 min
10.00	52.00±3.33a	0.00±0.00b	5.00±0.02a	0.00±0.00b	0.00±0.00b
15.00	65.00±5.00b	0.00±0.00b	3.00±0.33b	0.00±0.00b	0.00±0.00b
20.00	88.00±3.33c	0.00±0.00b	0.00±0.00b	0.00±0.00b	0.00±0.00b
25.00	90.00±2.89c	0.00±0.00b	0.00±0.00b	0.00±0.00b	0.00±0.00b
30.00	92.00±1.67c	0.00±0.00b	0.00±0.00b	0.00±0.00b	0.00±0.00b
初始含水量Initial moisture content	90.00±2.87c	0.00±0.00b	0.00±0.00b	0.00±0.00b	0.00±0.00b

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