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浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2019, Vol. 45 Issue (3): 296-305    DOI: 10.3785/j.issn.1008-9209.2018.07.041
园艺学     
鸢尾蒜种子无菌萌发及鳞茎形成
吕学思(),任梓铭,张栋,夏宜平()
浙江大学农业与生物技术学院园艺系，杭州 310058
Aseptic germination and bulblet formation of Ixiolirion tataricum (Pall.) Herb. seeds.
Xuesi Lü(),Ziming REN,Dong ZHANG,Yiping XIA()
Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China
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摘要：

以采自新疆的野生鸢尾蒜[Ixiolirion tataricum (Pall.) Herb.]种子为材料，比较研究了消毒方式、培养条件、种子预处理、贮藏条件对鸢尾蒜种子无菌萌发及离体鳞茎形成的影响。无菌播种结果表明：75%乙醇涡旋30 s，结合2% NaClO 5 min是种子的最佳消毒方式，污染率为0；鸢尾蒜种子在4 ℃条件下的萌发率在94.0%以上，而在25 ℃条件下则几乎不萌发。贮藏条件对种子萌发影响很大，4 ℃贮藏6个月后种子依旧能够正常萌发，而室温（25 ℃）贮藏6个月后种子完全丧失发芽力。筛选出的最有利于鸢尾蒜离体鳞茎形成的种子萌发培养基为MS+6-苄基腺嘌呤（6-benzylaminopurine, 6-BA）1.0 mg/L+萘乙酸（1-naphthylacetic acid, NAA）0.2 mg/L+蔗糖30 g/L，在该培养基上播种30 d后出芽整齐，然后转入25 ℃光照条件下，再转接至含60 g/L蔗糖的MS培养基上，可形成膨大的鳞茎。总之，该研究建立的鸢尾蒜无菌播种和鳞茎形成体系，为野生鸢尾蒜的离体资源保存与进一步开发利用提供了理论依据。
关键词： 鸢尾蒜;  无菌播种;  资源保存;  种子萌发;  鳞茎形成    
Abstract:

Ixiolirion tataricum (Pall.) Herb. is a unique and elite plant germplasm resource in China. The distribution of I. tataricum is limited in Xinjiang Uygur Autonomous Region. Ixiolirion. tataricum has strong resistance, high ornamental and medicinal values, which are of great potential in breeding. In this study, the seeds of I. tataricum collected from Xinjiang Uygur Autonomous Region were used as materials to compare the effects of sterilization method, culture conditions, gibberellin A3 (GA3) pretreatment and storage conditions on the aseptic germination and bulblet formation. The results showed that 30 s treatment with 75% ethanol followed by 5 min treatment with 2% NaClO was the best sterilization method for I. tataricum with 100% sterile seedlings. Using this sterilization method, the germination rate was more than 94.0% at 4 ℃, while almost no seed germinated at 25 ℃. Normally, seeds could germinate after storage for six months in the refrigerator at 4 ℃, but completely lost germination capacity at room temperature. We also found that MS+6-benzylaminopurine (6-BA) 1.0 mg/L+1-naphthylacetic acid (NAA) 0.2 mg/L+sucrose 30 g/L was the best germination medium for I. tataricum, which was beneficial to bulblet formation. Germinated in this medium, shoots could eventually expanded into bulbs after being transferred to light culture at 25 ℃ and then transferred to MS medium containing 60 g/L sucrose. This study provides an experimental basis for in vitro conservation of I. tataricum and to enhance the development and utilization of wild resources in China.
Key words: Ixiolirion tataricum (Pall.) Herb.    aseptic seeding    resource preservation    seed germination    bulblet formation
收稿日期: 2018-07-04 出版日期: 2019-06-25
CLC:  S 682.2  
基金资助: 浙江省“十三五”林木新品种选育(2016C02056-13);浙江省自然科学基金(LY17C150002)
通讯作者: 夏宜平     E-mail: lvxuesi2016@zju.edu.cn;ypxia@zju.edu.cn
作者简介: 吕学思（https://orcid.org/0000-0001-6425-3590），E-mail：lvxuesi2016@zju.edu.cn
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引用本文:

吕学思,任梓铭,张栋,夏宜平. 鸢尾蒜种子无菌萌发及鳞茎形成[J]. 浙江大学学报（农业与生命科学版）, 2019, 45(3): 296-305.

Xuesi Lü,Ziming REN,Dong ZHANG,Yiping XIA. Aseptic germination and bulblet formation of Ixiolirion tataricum (Pall.) Herb. seeds.. Journal of Zhejiang University (Agriculture and Life Sciences), 2019, 45(3): 296-305.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2018.07.041    或    http://www.zjujournals.com/agr/CN/Y2019/V45/I3/296

编号

Code

贮藏条件

Storage condition

预处理

Pretreatment

消毒流程

Sterilization flow

培养基

Medium

培养温度

Temperature/℃
S1 不贮藏 800 mg/L GA3 75%乙醇涡旋消毒30 s，结合0.1% HgCl2 3 min MS 4
S2 75%乙醇涡旋消毒30 s，结合0.1% HgCl2 5 min
S3 75%乙醇涡旋消毒30 s，结合0.1% HgCl2 8 min
S4 75%乙醇涡旋消毒30 s，结合2% NaClO 5 min
S5 75%乙醇涡旋消毒30 s，结合2% NaClO 10 min
S6 75%乙醇涡旋消毒30 s，结合2% NaClO 15 min
表1  消毒方式

编号

Code

贮藏条件

Storage condition

预处理

Pretreatment

消毒流程

Sterilization How

培养基

Medium

培养温度

Temperature/℃
T1 4 ℃，2个月 800 mg/L GA3

75%乙醇涡旋消毒30 s，

结合2% NaClO 5 min

 MS+6-BA 1.0 mg/L+NAA 0.2 mg/L 25
T2 4 ℃，2个月 800 mg/L GA3 MS+6-BA 1.0 mg/L+NAA 0.2 mg/L 4
T3 4 ℃，2个月 800 mg/L GA3 1/2 MS 25
T4 4 ℃，2个月 800 mg/L GA3 1/2 MS 4
T5 4 ℃，2个月 800 mg/L GA3 MS 25
T6 4 ℃，2个月 800 mg/L GA3 MS 4
T7 4 ℃，2个月 清水 MS 4
T8 25 ℃，2个月 800 mg/L GA3 MS 4
T9 25 ℃，2个月 清水 MS 4
T10 4 ℃，6个月 800 mg/L GA3 MS 4
T11 25 ℃，6个月 800 mg/L GA3 MS 4
表2  对种子的11种不同处理条件

编号

Code

污染率

Contamination rate/%

萌发时滞

Germination

time lag/d

萌发高峰期

Peak of

germination/d

发芽持续时间

Germination

time/d

萌发率

Germination rate/%

发芽势

Germination

energy/%

芽长

Shoot length/mm
S1 0 15.6±0.2a 17.4±0.4ab 6.8±0.2ab 96.0±2.4a 28.0±3.7c 7.25±0.82c
S2 0 16.4±0.5a 19.2±1.0a 8.8±1.5a 94.0±2.4a 32.0±4.9bc 5.25±1.02d
S3 0 16.4±0.2a 18.0±0.3ab 8.6±1.0a 94.0±2.4a 26.0±2.4c 3.95±0.77e
S4 0 13.8±0.4b 15.2±0.2c 3.2±0.7c 94.0±2.4a 52.0±8.0a 9.23±1.32b
S5 0 13.4±0.2b 15.2±0.2c 4.4±0.4bc 98.0±2.0a 48.0±5.8ab 9.89±0.66ab
S6 0 13.2±0.2b 15.0±0.0c 4.2±0.6c 96.0±4.0a 44.0±7.5abc 10.29±0.99a
表3  不同消毒方式下的种子萌发情况
图1  消毒方式对种子发芽的影响
图2   HgCl2 消毒条件下的种子在成苗阶段出现的细菌污染情况

编号

Code

萌发时滞

Germination

time lag/d

萌发高峰

Peak of

germination/d

发芽持续时间

Germination

time/d

萌发率

Germination

rate/%

发芽势

Germination

energy/%

芽长

Shoot length/

mm
T1 — — — 0.0±0.0b 0.0±0.0b —
T2 13.0±0.5a 14.6±0.5a 7.2±0.7a 94.0±4.0a 32.0±2.0a 5.35±0.82b
T3 — — — 4.0±2.4b 4.0±2.4b —
T4 13.0±0.5a 14.8±0.4a 4.6±0.5b 100.0±0.0a 42.0±3.7a 11.67±0.97a
T5 — — — 2.0±2.0b 2.0±2.0b —
T6 12.6±0.2a 13.0±0.0b 5.8±0.9ab 100.0±0.0a 38.0±3.7a 11.99±0.81a
表4  不同培养条件下的种子萌发情况
图3  培养条件对种子发芽的影响

编号

Code

萌发时滞

Germination

time lag/d

萌发高峰

Peak of

germination/d

发芽持续时间

Germination

time/d

萌发率

Germination rate/%

发芽势

Germination

energy/%

芽长

Shoot length/mm
T6 12.6±0.2b 13.0±0.0c 5.8±0.9b 100.0±0.0a 38.0±3.7a 11.99±0.81a
T7 13.8±0.6b 15.8±0.2b 5.6±0.7b 98.0±2.0ab 38.0±3.7a 10.65±1.00b
T8 15.4±0.4a 18.2±0.9a 17.0±2.6a 92.0±3.7ab 18.0±3.7b 5.19±1.59c
T9 16.8±0.6a 19.4±0.5a 15.0±3.6a 88.0±5.8b 18.0±3.7b 5.22±1.43c
表5  GA3与清水浸泡处理下的种子萌发情况

编号

Code

萌发时滞

Germination

time lag/d

萌发高峰

Peak of

germination/d

发芽持续时间

Germination

time/d

萌发率

Germination rate/%

发芽势

Germination

energy/%

芽长

Shoot length/mm
S4 13.8±0.4b 15.2±0.2b 3.2±0.7b 94.0±2.4ab 52.0±8.0a 9.23±1.32c
T6 12.6±0.2c 13.0±0.0c 5.8±0.9b 100.0±0.0a 38.0±3.7ab 11.99±0.81a
T8 15.4±0.4a 18.2±0.9a 17.0±2.6a 92.0±3.7b 18.0±3.7c 5.19±1.59d
T10 12.4±0.2c 13.8±0.4bc 5.2±0.7b 98.0±2.0ab 34.0±5.1bc 10.76±1.09b
T11 — — — 0.0±0.0c 0.0±0.0d —
表6  不同贮藏条件下的种子萌发情况
图4  转入25 ℃光照条件下培养的植株状态变化
图5  转入25 ℃光照条件下培养70 d后的植株状态
图6   100 d后不同培养基中植株的形态指标
图7  将植株转入MS培养基30 d后的鳞茎状态
1 钱啸虎 .中国鸢尾蒜属的研究.安徽师范大学学报(自然科学版),1980,4(1):157-158.
QIAN X H . A review of the Chinese species of Ixiolirion . Journal of Anhui Normal University (Natural Science), 1980,4(1):157-158. (in Chinese)
2 阎平 .新疆鸢尾蒜属的分类研究.石河子大学学报(自然科学版),1995(2):1-4.
YAN P . A study on taxonomy of genus Ixiolirion in Xinjiang. Journal of Shihezi University (Natural Science), 1995(2):1-4. (in Chinese with English abstract)
3 新疆植物志编辑委员会 .新疆植物志:第6卷.乌鲁木齐:新疆科技卫生出版社,1996:559-562.
Flora of Xinjiang Editorial Board . Flora of Xinjiang: vol.6. Urumqi: Xinjiang Science and Health Press, 1996:559-562. (in Chinese)
4 谢双全,田琳,阎平,等 .鸢尾蒜繁殖适应特征.江苏农业科学,2013,41(6):152-154.
XIE S Q , TIAN L , YAN P , et al . Reproductive adaptation characteristics of Ixiolirion . Jiangsu Agricultural Sciences, 2013,41(6):152-154. (in Chinese)
5 谢双全 .鸢尾蒜生物学特性及种子萌发特性的研究.乌鲁木齐:石河子大学,2013:31-39.
XIE S Q . A Study on biological characteristics of and seeds germination of Ixiolirion tataricum . Urumchi: Shihezi University, 2013:31-39. (in Chinese with English abstract)
6 CHANG C , CHEN C T , TSAI Y C , et al . A tissue culture protocol for propagation of a rare plant, Lilium speciosum Thunb. var. gloriosoides Baker. Botanical Bulletin of Academia Sinica, 2000,41(2):139-142.
7 任梓铭,夏宜平,李岳,等 .一种石蒜属植物外植体的消毒方法:CN105028197A.2015-11-11.
REN Z M , XIA Y P , LI Y , et al . A sterilization method of Lycoris explants: CN105028197A. 2015-11-11. (in Chinese)
8 田琳,田丽娜,谢双全,等 .准噶尔鸢尾蒜种子萌发特性研究.江苏农业科学,2012,40(5):364-366.
TIAN L , TIAN L N , XIE S Q , et al . Study on germination characteristics of Ixiolirion songaricum . Jiangsu Agricultural Sciences, 2012,40(5):364-366. (in Chinese)
9 田琳 .准噶尔鸢尾蒜形态结构及生物学特性研究.乌鲁木齐:石河子大学,2012:30-38.
TIAN L . Study of the morphological structure and biological characteristics of Ixiolirion songaricum . Urumqi: Shihezi University, 2012:30-38. (in Chinese with English abstract)
10 KLERK G J D . Micropropagation of bulbous crops: technology and present state. Floriculture and Ornamental Biotechnology, 2012(6):1-8.
11 朱宝疆,姜玉东 .植物生长物质与鳞茎形成之间的关系.科技致富向导,2011(21):223-224.
ZHU B J , JIANG Y D . Relationship of plant growth substances and bulb formation. Guide of Sci-tech Magazine, 2011(21):223-224. (in Chinese)
12 焦勇,孙杰英 .鸢尾蒜化学成分的研究(1).八一农学院学报,1988(1):83-87.
JIAO Y , SUN J Y . A study on chemical composition of Ixiolirion tataricum (Pall) Herb (I). Journal of August 1st Agricultural College, 1988(1):83-87. (in Chinese with English abstract)
13 黄文华,闫平,莫庸,等 .鸢尾蒜属植物过氧化物酶和酯酶同工酶研究.石河子大学学报(自然科学版),1995(3):27-32.
HUANG W H , YAN P , MO Y , et al . Studies on the isozymes of peroxidase and esterase of Ixiolirion . Journal of Shihezi University (Natural Science), 1995(3):27-32. (in Chinese with English abstract)
14 阎平,冯元忠,洪梅 .鸢尾蒜营养器官的解剖学研究.石河子大学学报(自然科学版),1994(3/4):1-6.
YAN P , FENG Y Z , HONG M . Atonomy research on vegetative organ of Ixiolirion tataricum . Journal of Shihezi University (Natural Science), 1994(3/4):1-6. (in Chinese with English abstract)
15 李岳,吴昀, ELMONGY M ,等 .野生郁金香种子无菌萌发及小鳞茎形成.浙江大学学报(农业与生命科学版),2017,43(3):307-316.
LI Y , WU Y , ELMONGY M , et al . Aseptic germination and bulblet formation of wild tulip (Tulipa sinkiangensi). Journal of Zhejiang University (Agriculture and Life Sciences), 2017,43(3):307-316. (in Chinese with English abstract)
16 苏家乐,李畅,陈璐,等 .不同预处理方法对牛皮杜鹃和小叶杜鹃种子萌发的影响.植物资源与环境学报,2011,20(4):64-69.
SU J L , LI C , CHEN L , et al . Effects of different pretreatment methods on seed germination of Rhododendron aureum and R . parvifolium. Journal of Plant Resources and Environment, 2011,20(4):64-69. (in Chinese with English abstract)
17 李娜,李子璇,曹小勇 .忽地笑种子萌发研究.种子,2012,31(10):18-21.
LI N , LI Z X , CAO X Y . Study on seed germination of L . aurea. Seed, 2012,31(10):18-21. (in Chinese with English abstract)
18 徐刚标 .植物种质资源离体保存研究进展.中南林学院学报,2000,20(4):81-87.
XU G B . Plant germplasm conservation in vitro . Journal of Central South Forestry University, 2000,20(4):81-87. (in Chinese with English abstract)
19 姚丽娟,杨燕萍,徐晓薇,等 .换锦花繁殖技术研究.北方园艺,2010(12):83-85.
YAO L J , YANG Y P , XU X W , et al . Study on the propagation technique of Lycoris sprengeri . Northern Horticulture, 2010(12):83-85. (in Chinese with English abstract)
20 彭俊翰 .马祖东引岛原生换锦花(石蒜科)之分类、生态与繁殖研究.中国，台北:台湾中兴大学,2013:110-115.
PENG J H . Studies of taxonomy, ecology and propagation of Lycoris sprengeri Comes ex Baker (Amaryllidaceae) Native to Dongyin, Matsu. Taipei， China: National Chung Hsing University, 2013:110-115. (in Chinese with English abstract)
21 王彩霞,欧阳彤,姜彦成 .新疆野生种与栽培种郁金香的杂交种子无菌萌发和鳞茎形成.植物生理学通讯,2009,45(11):1098-1100.
WANG C X , OUYANG T , JIANG Y C . Aseptic germination and bulblet formation of interspecific hybrids between wild species and cultivar of Tulip (Tulipa L.). Plant Physiology Communications, 2009,45(11):1098-1100. (in Chinese with English abstract)
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