全 文 :园 艺 学 报 2011,38(5):970–976 http: // www. ahs. ac. cn
Acta Horticulturae Sinica E-mail: yuanyixuebao@126.com
收稿日期:2010–10–08;修回日期:2011–03–21
基金项目:国家自然科学基金项目(30871730);教育部留学回国人员科研启动基金项目(教外司留[2009]8 号);中央高校基本科研业
务费专项
* 通信作者 Author for correspondence(E-mail:zhou2007@zju.edu.cn)
百合品种染色体倍性观察
李克虎 1,周桂雪 1,任贵玲 1,张线线 1,郭方其 2,周树军 1,*
(1 浙江大学农业与生物技术学院园艺系,农业部园艺植物生长发育与品质调控重点开放实验室,杭州 310058;
2浙江省农业科学院园艺研究所,杭州 310021)
摘 要:利用染色体常规制片技术分析了 45 个从荷兰引进的百合品种的染色体倍性,包括 21 个亚
洲百合、3 个麝香百合、7 个东方百合、8 个 LA 杂交品种、2 个 LO 杂交品种、4 个 OT 杂交品种。在 21
个亚洲百合中,12 个品种(‘Eleganza’、‘Monte Negro’、‘Renoir’、‘Black Bird’、‘Butter Pixie’、‘Orange
Matrix’、‘Petit Brigitte’、‘Pink Pixie’、‘Lollypop’、‘Red Latin’、‘Marrakech’和‘Pollyanna’)为二
倍体(2n = 2x = 24),5 个品种(‘Navona’、‘Orange Pixie’、‘Elodie’、‘Vermeer’和‘Lyon’)为三倍
体(2n = 3x = 36),4 个品种(‘Brunello’、‘Tresor’、‘Val di Sole’和‘Detroit’)为四倍体(2n = 4x = 48);
3 个麝香百合品种(‘White Heaven’、‘Gelria’和‘White Fox’)为二倍体(2n = 2x = 24);7 个东方百合
品种(‘Siberia’、‘Sorbonne’、‘Maru’、‘Dynamite’、‘Dordogne’、‘Montezuma’和‘Miss Lucy’)为二
倍体(2n = 2x = 24);8 个 LA 杂交品种(‘Brindis’、‘Courier’、‘Pavia’、‘Advantage’、‘Mestre’、‘Mombasa’、
‘Pirandello’和‘Serrada’)为三倍体(2n = 3x = 36);两个 LO 杂交品种(‘Triumphator’和‘White Triumph’)
为三倍体(2n = 3x = 36);4 个 OT 杂交品种中‘Yelloween’和‘Red Dutch’为二倍体(2n = 2x = 24),
‘Conca D’Or’和‘Golden City’为三倍体(2n = 3x = 36)。该结果表明,组内杂交品种(亚洲百合、东
方百合和麝香百合)中,除亚洲百合中有 5 个三倍体和 4 个四倍体品种外,其余 22 个为二倍体品种,因
而多数组内杂交品种为二倍体品种;而组间杂交品种(LA、OT 和 LO),只有 OT 中 2 个品种为二倍体,
其余 12 个皆为三倍体。
关键词:百合;品种;倍性
中图分类号:S 682.2 文献标识码:A 文章编号:0513-353X(2011)05-0970-07
Observation on Ploidy Levels of Lily Cultivars
LI Ke-hu1,ZHOU Gui-xue1,REN Gui-ling1,ZHANG Xian-xian1,GUO Fang-qi2,and ZHOU Shu-jun1,*
(1The Key Laboratory of Horticultural Plant Growth,Development and Quality Improvement,the Ministry of Agriculture
of the People’s Republic of China,Department of Horticulture,College of Agriculture and Biotechnology,Zhejiang
University,Hangzhou 310058,China;2Institute of Horticulture,Zhejiang Academy of Agricultural Sciences,Hangzhou
310021,China)
Abstract:45 lily cultivars imported from the Netherlands,including 21 Asiatic lilies,three
Longiflorum lilies,seven Oriental lilies,eight LA hybrid cultivars,two LO hybrid cultivars and four OT
hybrids cultivars,were analyzed using conventional chromosome preparation in this paper. Among the 21
5 期 李克虎等:百合品种染色体倍性观察 971
Asiatic cultivars,12 of them(‘Eleganza’,‘Monte Negro’,‘Renoir’,‘Black Bird’,‘Butter Pixie’,
‘Orange Matrix’,‘Petit Brigitte’,‘Pink Pixie’,‘Lollypop’,‘Red Latin’,‘Marrakech’,and
‘Pollyanna’)are diploid(2n = 2x = 24),five of them(‘Navona’,‘Orange Pixie’,‘Elodie’,‘Vermeer’
and‘Lyon’)are triploid(2n = 3x = 36),the rest four(‘Brunello’,‘Tresor’,‘Val di Sole’and‘Detroit’)
are tetraploid(2n = 4x = 48). All the three Longiflorum lilies(‘White Heaven’,‘Gelria’and‘White
Fox’)are diploid(2n = 2x = 24). All seven Oriental lilies(‘Siberia’,‘Sorbonne’,‘Maru’,‘Dynamite’,
‘Dordogne’,‘Montezuma’and‘Miss Lucy’)are also diploid(2n = 2x = 24). The eight LA hybrids
(‘Brindis’,‘Courier’,‘Pavia’,‘Advantage’,‘Mestre’,‘Mombasa’,‘Pirandello’and‘Serrada’)
are all triploid(2n = 3x = 36). Two LO hybrids(‘Triumphator’and‘White Triumph’)are also triploid
(2n = 3x = 36). Among the four OT hybrids,‘Yelloween’and‘Red Dutch’are diploid(2n = 2x = 24),
while‘Conca D’Or’and‘Golden City’are triploid(2n = 3x = 36). The results show that most of the lily
cultivars from intra-section hybridization(i.e.,Asiatic,Oriental and Longiflorum lilies)are diploid except
for five triploid and four tetraploid Asiatic lilies,by contrast,nearly all lily cultivars from inter-section
hybridization(i.e.,LA,OT and LO)are triploid except for two diploid OT cultivars.
Key words:lily;cultivar;ploidy
百合为重要的球根花卉,栽培品种多源于百合属的组内或组间杂交,其中组内杂交品种群主要
有亚洲百合(Asiatic lilies,A)、麝香百合(Longiflorum lilies,L)、喇叭百合(Trumpet lilies,T)
和东方百合(Oriental lilies,O),而组间杂交品种群有 LA(Longiflorum × Asiatic)、OT(Oriental ×
Trumpet)、LO(Longiflorum × Oriental)和 OA(Oriental × Asiatic)等。尽管我国有比较丰富的百
合野生资源(梁松筠 1995),但在新品种培育方面同荷兰相比相距甚远。培育具有自主知识产权的
百合品种是当前我国百合产业发展的一个重要方向。
在百合杂交育种中,知道亲本的染色体倍性对杂交成功与否非常重要。有关野生百合的染色体
倍性、常规核型及 C 带和以 rDNA 为探针的 FISH 核型已多有报道(Stewart,1947;图立古尔 等,
1996;Lim et al.,2001;王树红 等,2007;周树军 等,2008;胡凤荣 等,2009;刘冬云 等,2009;
荣立苹 等,2009)。这些报道表明除卷丹具有三倍体(2n = 3x = 36)的类群外,其余皆为二倍体(2n =
2x = 24)。然而,对百合栽培品种染色体倍性的报道仅限于一些二倍体品种(孙晓梅 等,2010)。
事实上,现在市场上的百合品种不仅数量多,而且既有二倍体,也有三倍体和四倍体,其中通过
麝香百合和亚洲百合杂交推出的品种多为三倍体(Zhou et al.,2008a)。由于百合染色体倍性从外
观上难以准确判断,因此,作者对目前国内市场出现的 45 个品种进行了倍性分析,供百合育种工
作参考。
1 材料与方法
1.1 材料来源
45 个百合品种均为从荷兰引进的品种,其中 21 个亚洲百合(A)品种、3 个麝香百合(L)
品种、7 个东方百合(O)品种、8 个 LA 杂交品种、2 个 LO 杂交品种和 4 个 OT 杂交品种。39
个品种购买自浙江虹越花卉有限公司,5 个由云南省农业科学院花卉研究所提供,1 个由北京农学
院提供(表 1)。
972 园 艺 学 报 38 卷
1.2 染色体制片
染色体分析采用常规压片法。待试管苗的根或温室栽培百合的茎生根约 1 ~ 3 cm 长时切下并用
蒸馏水清洗干净,再用 0.7 mmol · L-1 的放线菌酮室温预处理 4 h,然后用酒精︰乙酸(体积比 3︰1)
固定 12 h 后备用;将固定好的根尖用 1%的纤维素酶和果胶酶混合液软化后,用 2%醋酸洋红染色,
压片后用 Olympus(BH-2)观察,并对较好的中期分裂相照相以便对染色体进行计数。
2 结果与分析
在 21 个亚洲百合品种中,12 个品种(‘Eleganza’、‘Monte Negro’、‘Renoir’、‘Black Bird’、
‘Butter Pixie’、‘Orange Matrix’、‘Petit Brigitte’、‘Pink Pixie’、‘Lollypop’、‘Red Latin’、‘Marrakech’
和‘Pollyanna’)为二倍体(2n = 2x = 24),5 个品种(‘Navona’、‘Orange Pixie’、‘Elodie’、‘Vermeer’
和‘Lyon’)为三倍体(2n = 3x = 36),4 个品种(‘Brunello’、Tresor’、‘Val di Sole’和‘Detroit’)
为四倍体(2n = 4x = 48)。
3 个麝香百合(‘White Heaven’、‘Gelria’和‘White Fox’)都为二倍体(2n = 2x = 24)。
7 个东方百合品种(‘Siberia’、‘Sorbonne’、‘Maru’、‘Dynamite’、‘Dordogne’、‘Montezuma’
和‘Miss Lucy’)也都为二倍体(2n = 2x = 24)。
8 个 LA 杂交品种(‘Brindis’、‘Courier’、‘Pavia’、‘Advantage’、‘Mestre’、‘Mombasa’、
‘Pirandello’和‘Serrada’)皆为三倍体(2n = 3x = 36)。
两个 LO 杂交品种(即:‘Triumphator’和‘White Triumph’)都为三倍体(2n = 3x = 36)。
4 个 OT 杂交品种中‘Yelloween’和‘Red Dutch’为二倍体(2n = 2x = 24),‘Conca D’Or’和
‘Golden City’为三倍体(2n = 3x = 36)(表 1,图 1)。
该结果表明,组内品种(亚洲百合、东方百合和麝香百合)中,除亚洲百合中有三倍体和四倍
体品种,其余均为二倍体品种,因而组内品种主要为二倍体品种;然而,组间杂交品种(LA、OT
和 LO),只有 OT 百合的‘Yelloween’和‘Red Dutch’为二倍体品种,其余皆为三倍体品种,因此,
组间杂交品种三倍体占绝对优势。
3 讨论
由于不同组百合的农艺性状具有较大差异,如:亚洲百合花色丰富,花朵朝上,较抗病毒和枯
萎病;东方百合香气甜美,较抗灰霉病;麝香百合易于促成栽培,因此,培育组间杂交品种是百合
育种的一个重要方向。同时,由于多倍体同二倍体相比较在器官大小和生长势方面具有较大的优势,
因而,百合多倍体育种也是重要的方向。
百合组间杂交的 F1 代通常由于减数分裂不正常而高度不育(Zhou et al.,2008b),但有些可以
产生少量体细胞配子(2n 配子),从而产生三倍体的回交一代,这一过程称作有性多倍化
(Barba-Gonzalez et al.,2004,2005a,2005b),用基因组荧光原位杂交技术可证明荷兰百合育种公
司通过有性多倍化从回交一代中筛选了很多三倍体 LA 品种(Zhou et al.,2008a)。因此有理由相信
本试验中的 LA、OT 和 LO 三倍体品种可能主要来源于有性多倍化。
当然,本试验中的有些三倍体也可能来源于无性多倍化的途径,即:在百合育种中通常用安磺
灵对不育的 F1 代加倍使其恢复育性,然后再与二倍体回交获得三倍体(van Tuyl,1989;van Tuyl et
al.,1992)。
有性多倍化比无性多倍化有更多的优势:第一,有性多倍化大大缩短了百合的三倍体育种年限;
5 期 李克虎等:百合品种染色体倍性观察 973
第二,有性多倍化中 F1 产生的体细胞配子有基因组间染色体重组,从而大大增加了 2n 配子变异,
提高了后代筛选的机会(Ramanna & Jacobsen,2003)。这也正是市场上有较多三倍体组间杂交品种
的原因。
表 1 供试百合品种
Table 1 Lily cultivars used in this study
编号 Code 类型 Type 品种 Cultivar 来源 Origin 倍性 Ploidy
1 亚洲百合 Asiatic lilies Eleganza 虹越 Hongyue 2n = 2x = 24
2 Monte Negro 虹越 Hongyue 2n = 2x = 24
3 Renoir 虹越 Hongyue 2n = 2x = 24
4 Black Bird 虹越 Hongyue 2n = 2x = 24
5 Butter Pixie 虹越 Hongyue 2n = 2x = 24
6 Orange Matrix 虹越 Hongyue 2n = 2x = 24
7 Petit Brigitte 虹越 Hongyue 2n = 2x = 24
8 Pink Pixie 虹越 Hongyue 2n = 2x = 24
9 Lollypop 虹越 Hongyue 2n = 2x = 24
10 Red Latin 虹越 Hongyue 2n = 2x = 24
11 Marrakech 云南 Yunnan 2n = 2x = 24
12 Pollyanna 云南 Yunnan 2n = 2x = 24
13 Vermeer 虹越 Hongyue 2n = 3x = 36
14 Lyon 云南 Yuannan 2n = 3x = 36
15 Navona 虹越 Hongyue 2n = 3x = 36
16 Orange Pixie 虹越 Hongyue 2n = 3x = 36
17 Elodie 虹越 Hongyue 2n = 3x = 36
18 Tresor 虹越 Hongyue 2n = 4x = 48
19 Brunello 虹越 Hongyue 2n = 4x = 48
20 Val di Sole 虹越 Hongyue 2n = 4x = 48
21 Detroit 虹越 Hongyue 2n = 4x = 48
22 麝香百合 Longiflorum lilies White Heaven 虹越 Hongyue 2n = 2x = 24
23 Gelria 云南 Yunnan 2n = 2x = 24
24 White Fox 云南 Yunnan 2n = 2x = 24
25 东方百合 Oriental lilies Siberia 虹越 Hongyue 2n = 2x = 24
26 Sorbonne 虹越 Hongyue 2n = 2x = 24
27 Maru 虹越 Hongyue 2n = 2x = 24
28 Dynamite 虹越 Hongyue 2n = 2x = 24
29 Dordogne 虹越 Hongyue 2n = 2x = 24
30 Montezuma 虹越 Hongyue 2n = 2x = 24
31 Miss Lucy 虹越 Hongyue 2n = 2x = 24
32 LA 百合 LA lilies Brindis 虹越 Hongyue 2n = 3x = 36
33 Courier 虹越 Hongyue 2n = 3x = 36
34 Pavia 虹越 Hongyue 2n = 3x = 36
35 Advantage 虹越 Hongyue 2n = 3x = 36
36 Mestre 虹越 Hongyue 2n = 3x = 36
37 Mombasa 虹越 Hongyue 2n = 3x = 36
38 Pirandello 虹越 Hongyue 2n = 3x = 36
39 Serrada 虹越 Hongyue 2n = 3x = 36
40 LO 百合 LO lilies Triumphator 虹越 Hongyue 2n = 3x = 36
41 White Triumph 虹越 Hongyue 2n = 3x = 36
42 OT 百合 OT lilies Yelloween 虹越 Hongyue 2n = 2x = 24
43 Conca DOr 虹越 Hongyue 2n = 3x = 36
44 Red Dutch 虹越 Hongyue 2n = 2x = 24
45 Golden City 北京 Beijing 2n = 3x = 36
974 园 艺 学 报 38 卷
5 期 李克虎等:百合品种染色体倍性观察 975
图 1 百合 45 个品种的根尖细胞分裂中期染色体
1 ~ 21. 亚洲百合,其中 1 ~ 12 为二倍体,13 ~ 17 为三倍体,18 ~ 21 为四倍体;22 ~ 24. 麝香百合,为二倍体;25 ~ 31.东方百合,为二倍
体;32 ~ 39. LA 百合,为三倍体;40、41. LO 百合,为三倍体;42 ~ 45. OT 百合,42 和 44 为二倍体,43 和 45 为三倍体。
图 1 中编号与表 1 中编号相同。
Fig. 1 The chromosomes at metaphase of root tip cells of 45 lily cultivars
1–21:Asiatic lilies. Among them,1–12 are diploid,13–17 triploid and 18–21 tetraploid;22–24:Longiflorum lilies. All the three cultivars are
diploid;25–31:Oriental lilies. All of them are also diploid;32–39:LA lilies. All of them are triploid;40,41:LO lilies.
Both of them are triploid;42–45:OT lilies. Among them,42 and 44 are diploid,43 and 45 are triploid.
The number of each plate is correspondent to each cultivar code showed in table 1.
976 园 艺 学 报 38 卷
References
Barba-Gonzalez R,Lim K B,Ramanna M S,Visser R G F,van Tuyl J M. 2005a. Occurrence of 2n gametes in the F1 hybrids of Oriental × Asiatic
lilies(Lilium):Relevance to intergenomic recombination and backcrossing. Euphytica,143:67–73.
Barba-Gonzalez R,Lim K B,Ramanna M S,Visser R G F,van Tuyl J M. 2005b. The occurrence of intergenomic recombination in the F1 hybrids
of Oriental × Asiatic lily hybrids(Lilium)and its significance for genetic variation in the BC1 progenies as revealed by GISH and FISH analyses.
Genome,48:884–894.
Barba-Gonzalez R,Lokker B H,Lim K B,Ramanna M S,van Tuyl J M. 2004. Use of 2n gametes for the production of sexual polyploids from sterile
Oriental × Asiatic hybrids of lilies(Lilium). Theor Appl Genet,109:1125–1132.
Hu Feng-rong,Liu Guang-xin,Luo Feng-xia,Xi Meng-li,Xu Jin,Shi Ji-sen. 2009. Identification of Lilium hybrid by Giemsa C-banding and 45S
rDNA FISH. Journal of Nanjing Forestry University:Nat Sci Ed,33:30–34. (in Chinese)
胡凤荣,刘光欣,罗凤霞,席梦利,徐 进,施季森. 2009. 利用 Giemsa C–带和 45S rDNA FISH 的方法鉴定百合杂种. 南京林业大学
学报:自然科学版,33:30–34.
Liang Song-jun. 1995. Chorology of Liliaceae(s.str.)and its bearing on the Chinese flora. Acta Phytotaxonomica Sinica,33:27–51. (in Chinese)
梁松筠. 1995. 百合科(狭义)植物的分布区对中国植物区系研究的意义. 植物分类学报,33:27–51.
Lim K B,Wennekes J,De Jong J H,Jacobsen E,van Tuyl J M. 2001. Karyotype analysis of Lilium longiflorum Thunb and Lilium rubellum Baker
by chromosome banding and fluorescence in situ hybridization. Genome,44:911–918.
Liu Dong-yun,Zhang Xiao-man,Yang Nan,Zhang Li-juan,Shao Shuai. 2009. Study on karyotypes of Lilium davidii var. unicolor and Lilium
concolor var. buschianum Baker. Journal of Northwest A & F University:Nat Sci Ed,37:102–106. (in Chinese)
刘冬云,张晓曼,杨 楠,张利娟,邵 帅. 2009. 兰州百合和有斑百合的核型研究. 西北农林科技大学学报:自然科学版,37:102–
106.
Ramanna M S,Jacobsen E. 2003. Relevance of sexual polyploidization for crop improvement–A review. Euphytica,133:3–18.
Rong Li-ping,Lei Jia-jun,Zheng Yang,Gao Yu-fu. 2009. Study on karyotypes of Lilium species native to northeast China. Journal of Jilin
Agricultural University,31(6):711–716. (in Chinese)
荣立苹,雷家军,郑 洋,高玉福. 2009. 东北地区野生百合的核型研究. 吉林农业大学学报,31 (6):711–716.
Stewart R N. 1947. The morphology of somatic chromosomes in Lilium. Amer J Bot,34:9–26.
Sun Xiao-mei,Jia Lian,Yang Hong-guang,Zhang Li-jie,Cui Wen-shan. 2010. Three cultivars karyotype analysis of Lilium × formolongi. Acta
Horticulturae Sinica,37 (6):1003–1008. (in Chinese)
孙晓梅,贾 莲,杨宏光,张丽杰,崔文山. 2010. 新铁炮百合 3 个品种的核型分析. 园艺学报,37 (6):1003–1008.
Tolgor,Liu Li-bo. 1996. Studies of karyotypes of 5 species in Lilium from Jilin. Journal of Wuhan Botanical Research,14 (1):6–12. (in Chinese)
图立古尔,刘立波. 1996. 吉林产 5 种百合的核型研究. 武汉植物学研究,14 (1):6–12.
van Tuyl J M. 1989. Research on mitotic and meiotic polyploidization in lily breeding. Herbertia,45:97–103.
van Tuyl J M,Meijer H,van Diën M P. 1992. The use of oryzalin as an alternative for colchicine in vitro chromosome doubling of Lilium and Nerine.
Acta Hort,325:625–630.
Wang Shu-hong,Li Qiao-xia,Zhao Qing-fang,Liang Wan-fu. 2007. The karyotypes analysis of four ornamental Lilium. Journal of Northwest
Normal University:Nat Sci Ed,43 (4):93–97. ( in Chinese)
王树红,李巧峡,赵庆芳,梁万福. 2007. 四种观赏百合的核型分析. 西北师范大学学报:自然科学版,43 (4):93–97.
Zhou S,Ramanna M S,Visser R G F,van Tuyl J M. 2008a. Genome composition of triploid lily cultivars derived from sexual polyploidization of
Longiflorum × Asiatic hybrids(Lilium). Euphytica,160:207–215.
Zhou S,Ramanna M S,Visser R G F,van Tuyl J M. 2008b. Analysis of the meiosis in the F1 hybrids of Longiflorum × Asiatic(LA)of lilies(Lilium)
using genomic in situ hybridization. Journal of Genetics and Genomics,35:687–695.
Zhou Shu-jun,van Tuyl J M,Zang De-kui,Xia Yi-ping,Li Fang. 2008. Physical localization of 45S rDNA on the chromosomes of 4 species of the
genus Lilium. Acta Horticulturae Sinica,35 (6):859–862. (in Chinese)
周树军,van Tuyl J M,臧德奎,夏宜平,李 方. 2008. 45S rDNA 在 4 种百合属植物染色体上的物理定位. 园艺学报,35 (6):859–862.