超低温保存下藓类植物孢子萌发率的变化：以6种藓类植物为例

孙莉玮; 高雪笛; 许丹尔; 王书琴; 杨卓筠; 王健

doi:10.3969/j.issn.1000-5641.2019.03.015

超低温保存下藓类植物孢子萌发率的变化：以6种藓类植物为例

doi: 10.3969/j.issn.1000-5641.2019.03.015

华东师范大学生命科学学院, 上海 200241

基金项目:

国家自然科学基金 31770371

华东师范大学本科生创业训练培育项目 2017PY-424

上海市大学生创新活动项目 201510269105

详细信息

作者简介:
孙莉玮, 女, 硕士研究生, 研究方向为苔藓植物分子地理及种质资源保存研究.E-mail:51161300063@stu.ecnu.edu.cn

通讯作者:
王健, 男, 副研究员, 主要从事苔藓植物分类及种质资源保存研究.E-mail:jwang@bio.ecnu.edu.cn

中图分类号: Q94.331
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出版历程
- 收稿日期: 2018-05-25
- 刊出日期: 2019-05-25

Variation in germination rates of moss spores using a cryopreservation technique: A case study of spores from six moss species

School of Life Sciences, East China Normal University, Shanghai 200241, China

摘要

摘要: 为了检验苔藓植物孢子超低温保存方法的适用性，该文对采自不同生境中的6种藓类孢子超低温保存前的干燥时间和低温预处理进行探索，并在最适处理条件下对其中5种藓类孢子进行不同时间梯度（1 d、15d、30 d、90 d和180 d）的超低温保存研究.结果表明：①经过干燥和低温预处理后，6种藓类孢子的最高萌发率均维持在较高水平（87.25%~96.21%）；②与低温预处理相比，干燥处理在孢子超低温保存中更为关键.孢子在只进行干燥处理情况下其最高平均萌发率为74.97%~96.21%，而只对其进行低温预处理时的最高平均萌发率变化较大，从21.73%到90.94%；③5种藓类孢子在液氮保存1d后的平均萌发率均最高（87.25%~96.21%），且随着保存时间的延长，所有藓类孢子的萌发率都有所下降，但仍维持在相对较高的水平（73.69%~86.60%）；④葫芦藓、丝瓜藓和长蒴藓孢子在保存30d后萌发率分别为86.49%、86.60%和84.98%，卵蒴真藓和丛生真藓孢子在保存180d后萌发率仍可达到73.69%和84.17%.研究表明，藓类孢子非常适合于超低温保存，而且可能代表一种简单、稳定、高效的苔藓植物种质资源保存材料.
- 苔藓植物 /
- 藓类 /
- 生境 /
- 孢子 /
- 超低温保存
Abstract: To evaluate the applicability of cryopreservation for bryophyte spores, this study explored the effect of dehydration time and low temperature pretreatment; the study was used to identify the optimal conditions for six moss spores collected from different habitats. In addition, varying time periods (1 d, 15 d, 30 d, 90 d, 180 d) of spore cryopreservation were studied under the optimal pretreatment conditions identified. The results showed that:①After dehydration and low temperature pretreatment, the highest germination rates of spores from the six moss species were maintained (87.25%~96.21%). ②Compared to low temperature pretreatment, dehydration seems to be the key point of success for spore cryopreservation. The average spore germination rates ranged from 74.97% to 96.21% when treated with dehydration alone; in comparison, germination rates ranged from 21.73% to 90.94% when pretreated with low temperature alone. ③The average germination rates of spores from five moss species maintained their respective highest level (87.25%~96.21%) after cryopreservation for 1 d and then decreased with the prolongation of storage time; nevertheless, the germination rates still remained at relatively high levels (73.69%~86.60%).④ The average germination rates of spores from Funaria hygrometrica, Pohlia elongate, and Trematodon longicollis after storage for 30 d were 86.49%, 86.60%, and 84.98%, respectively. The germination rates of spores from Bryum blindii and B. caespiticium reached 73.69% and 84.17%, respectively, after storage for 180 d. The study indicates that the procedure for spore cryopreservation could be a simple, stable, and efficient method for storage of bryophyte species in conservation programs.
- bryophyte /
- moss /
- habitats /
- spores /
- cryopreservation

HTML全文

图 1 不同干燥时间和低温预处理条件下6种藓类孢子超低温保存后萌发率比较

注: (a)长蒴藓; (b)葫芦藓; (c)卵蒴真藓; (d)丛生真藓; (e)丝瓜藓; (f)日本曲尾藓.萌发率用平均值和标准差表示, 字母不同表示差异显著($\alpha $=0.05)

Fig. 1 A comparison of germination rates for spores from six moss species after cryopreservation with varying dehydration times and low temperature pretreatments

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表 1 实验材料

Tab. 1 Materials used in experiment

种名	采集地	采集年月	生境	海拔/m
卵蒴真藓Bryum blindii Bruch & Schimp.	四川康定	2014.07	土生(湿润)	2 993
丛生真藓Bryum caespiticium Hedw.	贵州凯里	2014.07	土生(干旱)	2 594
日本曲尾藓Dicranum japonicum Mitt.	西藏上察隅	2013.05	朽木生(湿润)	1 919
葫芦藓Funaria hygrometrica Hedw.	上海华东师范大学	2014.04	土生(干旱)	35
丝瓜藓Pohlia elongata Hedw.	云南丽江	2014.07	朽木生(湿润)	3 000
长蒴藓Trematodon longicollis Michx.	广西上思	2014.04	土生(干燥)	185

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表 2 5种藓类孢子超低温保存不同时间后萌发率比较

Tab. 2 Comparison of germination rates for spores from five moss species under different

保存时间/d	平均萌发率/%
保存时间/d	长蒴藓 T. longicollis	葫芦藓 F. hygrometrica	丝瓜藓 P. elongata	卵蒴真藓 B. blindii	从生真藓 B. caespiticium
1	96.21$\pm $2.4$^{a}$	94.31$\pm $3.27$^{a}$	90.86$\pm $3.75$^{a}$	87.25$\pm $1.53$^{a}$	92.01$\pm $4.08$^{a}$
	(103.84)	(102.44)	(229.62)	(170.4)	(128.09)
15	93.17$\pm $4.31$^{a}$	77.03$\pm $11.25$^{b}$	76.97$\pm $3.37$^{b}$	53.19$\pm $0.88$^{d}$	79.56$\pm $10.90$^{b}$
	(100.55)	(83.66)	(194.52)	(102.87)	(106.32)
30	84.98$\pm $0.90$^{b}$	86.49$\pm $5.89$^{ab}$	86.60$\pm $3.03$^{a}$	78.84$\pm $3.66$^{b}$	89.72$\pm $1.14$^{ab}$
	(91.72)	(93.94)	(218.85)	(153.96)	(119.90)
90	—	—	—	62.39$\pm $4.38$^{c}$	71.79$\pm $5.66$^{b}$
				(121.83)	(95.94)
180	—	—	—	73.69$\pm $3.44$^{b}$	84.17$\pm $3.30$^{ab}$
				(143.89)	(112.48)
注: —表示缺乏实验数据.萌发率用平均值$\pm $标准差表示; 括号中的数值为相对保持率, 即LN保存后的平均萌发率/LN保存前的平均萌发率; 5种藓类孢子独立进行萌发率差异显著性比较, 同列字母不同者表示差异显著($\alpha $=0.05)

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