观赏百合种质资源多样性研究
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摘要
观赏百合因其植株花形大、端庄飘逸，是切花中的优良品种，深受广大消费者的青睐，是目前国际市场上十分畅销的花卉之一，具有广阔的市场开发前景。
全球已发现百合共有96种左右，主要分布在中国、目本、美国和欧洲等温带地区。我国约分布有46种左右，是世界百合的分布中心。但是由于种种原因，我国在百合的育种方面非常薄弱，目前还没有培育出比较知名的杂交品种或品系，当前国内商业栽培种球主要依赖于国外进口。
本文对来自亚洲系、东方系和铁炮杂种系的一些著名品种进行分子标记、花粉形态扫描及组织培养的多样性研究，再结合已有的资料进行综合分析，旨在了解三种品系百合的遗传多样性状况、育种背景及相互间亲缘关系，为我国观赏百合的育种工作提供一些基础性资料。
用15个引物对三个品系来源的15个百合品种进行扩增，15个引物扩增出218个条带，其中品种间多态性条带为216条，占总扩增带数的99.08％。三个杂种系中，亚洲系各品种的多态性条带比率(PPB)为40.83％，Nei's基因多样度(h)为0.1654，Shannon多样性指数(Ⅰ)为0.2415；东方系各品种的PPB为42.20％，h为0.1600，Ⅰ为0.2381；铁炮系各品种的PPB为25.23％，h为0.1072，Ⅰ为0.1551。三个杂种百合品系中，铁炮系品种的遗传多样性水平最低，亚洲系品种的遗传多样性水平最高。
品种间Nei's氏遗传距离系数显示：不同品系百合品种之间的遗传距离明显大于同一品系百合品种之间的遗传距离，反映了同一品系来源的百合品种具有较近的亲缘关系，而不同品系来源的百合品种具有较远的亲缘关系。三个杂种系中，东方系各品种和亚洲系各品种之间的遗传距离较小，亲缘关系最近；铁炮系各品种与亚洲系各品种之间的亲缘关系最远。
应用扫描电镜对三个杂种系百合的部分品种和兰州百合、细叶百合的花粉进行形状、大小、外壁纹饰等方面的比较研究，结果显示：百合花粉的形状皆呈椭球体，具单沟萌发孔，沟长达至两端，花粉外壁皆为网状纹饰。不同品种、品系来源的百合花粉粒在大小、P／E值和外壁纹饰上存在不同程度的差异，对区分某些品种、品系和品种、品系之间亲缘关系的鉴定具有一定的参考价值。其中同一品系来源的百合品种，其花粉的主要特征相似性较大；不同来源的百合品种，其花粉主要特征差异显著。东方系品种的P／E值在0.375～0.405之间，亚洲系品种P／E值在0.401～0.402之间，铁炮系品种的P／E值在0.450～0.507之间。三个杂种系中，铁炮系品种的P／E最大，东方系和亚洲系品种的P／E较为接近。花粉表面纹饰显示：三种杂种系之间，东方系和亚
洲系百合纹饰较相似，网脊上都有瘤状或疵状颗粒，而铁炮系列百合网脊平滑，无颗
粒排布。反映了东方系和亚洲系百合具有较近的亲缘关系或遗传背景。
对三个品系中的部分常见品种进行组织培养及试管苗快速繁殖的研究。结果显
示:初代鳞片诱导及芽的增殖过程中，丛生芽和小鳞茎的形成率、生成数目及其生长
情况与百合基因型之间有很大的相关性，不同品系来源的百合品种在培养过程中表现
出较大的差异，而同一品系来源的百合品种则表现出较多的相似。
RAPD标记结果显示:花色、花型和百合品系遗传多样性、品种间亲缘关系有很
大的相关性，可以用来粗略评价百合品系的遗传多样性状况，鉴定品系内各品种间的
亲缘关系。RAPD分析涉及的品种中，亚洲系品种和东方系品种的花型、花色具多样
化;铁炮系品种的花色、花型非常单一。铁炮系百合的遗传多样性水平最低，东方系
和亚洲系百合的遗传多样性水平较高。品种间Nei，S氏遗传距离显示:同一品系中，
相同花色的百合品种具有较小的遗传距离;不同花色的百合品种其遗传距离也较大。
主要研究结论如下:
1.三个杂种系中，亚洲系和东方系百合遗传多样性水平较高，铁炮系百合遗传多样
性水平最低;
2.亚洲系和东方系百合品种的亲缘关系较近;
3.同一杂种系来源的百合品种在花粉形态、组织培养与植株再生等方面具有很高的
相似性;
4.花色、花型和百合品系遗传多样性、品种间亲缘关系有很大的相关性。
Lilium is a great genus which includes abundance species and cultivars. Lilium is also one of the important ornamentals crops and ranks seventh among the cut flowers auctioned in the world. Because of the glorious blooms and scent, they were mass-produced as potted plant, cut flower and courtyard plant.
China is the distribution centre of lilium .There are almost 46 out of 96 species in China. While for some reasons, the breeding of lily is undeveloped. At present, there are not famous varieties and strains.which are breeded in china. The bulbs which have been cultured for trade chiefly depend on import.
The Asiatic Oriental and Longiflorum hybrids of Lilium are the world-wide hybrid strains , which are largely traded in the international market. In the present study, diversity of RAPD markers, pollen character and micropropagation of lily cultivars were studied. By combination of results in the present study with others that have been conducted in other previous research and analysis of diversity, breeding background of the above three strains , relationships among the cultivars, some conclusion expected to be drawed for the breeding of ornamentals lily in China.
Based on 15 random primers , RAPD polymorphism of 15 lilium cultivars were detected. Of 218 bands, 216 were polymorphic (99.08%) . The PPB, h and /of the Asiatic cultivars were 40.83%, 0.1654, 0.2415, respectively; the PPB, h and I of the Oriental cultivars were 42.20%, 0.1600, 0.2381, respectively; the PPB, h and /of the Longiflorum cultivars were 25.23%, 0.1072, 0.1551 respectively. The lily cultivars of the Asiatic hybrids had the highest genetic diversity; Longiflorum hybrids had the lowest genetic diversity.
Neis genetic distance among the cultivars showed that value of gernetic distance among the cultivars from the same strain were lower than that from different strains. The results revealed that cultivars from the same strain had much closer relationships; the cultivars from different strains had less close relationships. Among these three strains, gernetic distance between Asiatic cultivars and Oriental cultivars was the lowest, which was the highest between the Asiatic cultivars and Longiflorum cultivars. The Oriental strain and Asiatic strain had relatively closer relationship.
The pollen grains of some cultivars from the three hybrid strains were examined under
scanning electron microscope (SEM). The pollen character and gross morphology in Lilium were discussed: The pollen grains were generally elliptic and monocolpate(distal). Evidence from pollen morphology indicated that the pollen size, P/E value and exine sculpturing among species, cultivars were different in varying degrees, which could be used to distinguish cultivars. The main character of cultivars from the same strain are similar, which revealed that these cultivars had closer relationships. The exine sculpturing also revealed that there was more similarity between Asiatic Hybrids and Oriental Hybrids.
In present study, The difference in clonal propagation of the three strains cultivars were discussed through culture of scales. The induction proportion and the growth status of the buds and little bulbs, the proliferation of the buds and their growth status had direct correlation on genotype of lily. Cultivars from the same hybrid strain had more similarity .
The study showed the flower colour and shape were related with genetic diversity and relationships of lily cultivars. The Oriental and Asiatic hybrid cultivars were rich in flower colour and shape, they also had high genetic diversity. The Longiflorum hybrid cultivars was poor in flower colour and shape, they also had lower genetic diversity. In the same strain, cultivars that were same in flower colour were more familiar than others. The above results could be used to distinguish the relationship of lily cultivars from the same strains.
The main results are as follows:
1. The lily cultivars of the Asiatic hybrids strains had the highest genetic diversity; Longiflorum hybrid strains had the lowest gene
全球已发现百合共有96种左右，主要分布在中国、目本、美国和欧洲等温带地区。我国约分布有46种左右，是世界百合的分布中心。但是由于种种原因，我国在百合的育种方面非常薄弱，目前还没有培育出比较知名的杂交品种或品系，当前国内商业栽培种球主要依赖于国外进口。
本文对来自亚洲系、东方系和铁炮杂种系的一些著名品种进行分子标记、花粉形态扫描及组织培养的多样性研究，再结合已有的资料进行综合分析，旨在了解三种品系百合的遗传多样性状况、育种背景及相互间亲缘关系，为我国观赏百合的育种工作提供一些基础性资料。
用15个引物对三个品系来源的15个百合品种进行扩增，15个引物扩增出218个条带，其中品种间多态性条带为216条，占总扩增带数的99.08％。三个杂种系中，亚洲系各品种的多态性条带比率(PPB)为40.83％，Nei's基因多样度(h)为0.1654，Shannon多样性指数(Ⅰ)为0.2415；东方系各品种的PPB为42.20％，h为0.1600，Ⅰ为0.2381；铁炮系各品种的PPB为25.23％，h为0.1072，Ⅰ为0.1551。三个杂种百合品系中，铁炮系品种的遗传多样性水平最低，亚洲系品种的遗传多样性水平最高。
品种间Nei's氏遗传距离系数显示：不同品系百合品种之间的遗传距离明显大于同一品系百合品种之间的遗传距离，反映了同一品系来源的百合品种具有较近的亲缘关系，而不同品系来源的百合品种具有较远的亲缘关系。三个杂种系中，东方系各品种和亚洲系各品种之间的遗传距离较小，亲缘关系最近；铁炮系各品种与亚洲系各品种之间的亲缘关系最远。
应用扫描电镜对三个杂种系百合的部分品种和兰州百合、细叶百合的花粉进行形状、大小、外壁纹饰等方面的比较研究，结果显示：百合花粉的形状皆呈椭球体，具单沟萌发孔，沟长达至两端，花粉外壁皆为网状纹饰。不同品种、品系来源的百合花粉粒在大小、P／E值和外壁纹饰上存在不同程度的差异，对区分某些品种、品系和品种、品系之间亲缘关系的鉴定具有一定的参考价值。其中同一品系来源的百合品种，其花粉的主要特征相似性较大；不同来源的百合品种，其花粉主要特征差异显著。东方系品种的P／E值在0.375～0.405之间，亚洲系品种P／E值在0.401～0.402之间，铁炮系品种的P／E值在0.450～0.507之间。三个杂种系中，铁炮系品种的P／E最大，东方系和亚洲系品种的P／E较为接近。花粉表面纹饰显示：三种杂种系之间，东方系和亚
洲系百合纹饰较相似，网脊上都有瘤状或疵状颗粒，而铁炮系列百合网脊平滑，无颗
粒排布。反映了东方系和亚洲系百合具有较近的亲缘关系或遗传背景。
对三个品系中的部分常见品种进行组织培养及试管苗快速繁殖的研究。结果显
示:初代鳞片诱导及芽的增殖过程中，丛生芽和小鳞茎的形成率、生成数目及其生长
情况与百合基因型之间有很大的相关性，不同品系来源的百合品种在培养过程中表现
出较大的差异，而同一品系来源的百合品种则表现出较多的相似。
RAPD标记结果显示:花色、花型和百合品系遗传多样性、品种间亲缘关系有很
大的相关性，可以用来粗略评价百合品系的遗传多样性状况，鉴定品系内各品种间的
亲缘关系。RAPD分析涉及的品种中，亚洲系品种和东方系品种的花型、花色具多样
化;铁炮系品种的花色、花型非常单一。铁炮系百合的遗传多样性水平最低，东方系
和亚洲系百合的遗传多样性水平较高。品种间Nei，S氏遗传距离显示:同一品系中，
相同花色的百合品种具有较小的遗传距离;不同花色的百合品种其遗传距离也较大。
主要研究结论如下:
1.三个杂种系中，亚洲系和东方系百合遗传多样性水平较高，铁炮系百合遗传多样
性水平最低;
2.亚洲系和东方系百合品种的亲缘关系较近;
3.同一杂种系来源的百合品种在花粉形态、组织培养与植株再生等方面具有很高的
相似性;
4.花色、花型和百合品系遗传多样性、品种间亲缘关系有很大的相关性。
Lilium is a great genus which includes abundance species and cultivars. Lilium is also one of the important ornamentals crops and ranks seventh among the cut flowers auctioned in the world. Because of the glorious blooms and scent, they were mass-produced as potted plant, cut flower and courtyard plant.
China is the distribution centre of lilium .There are almost 46 out of 96 species in China. While for some reasons, the breeding of lily is undeveloped. At present, there are not famous varieties and strains.which are breeded in china. The bulbs which have been cultured for trade chiefly depend on import.
The Asiatic Oriental and Longiflorum hybrids of Lilium are the world-wide hybrid strains , which are largely traded in the international market. In the present study, diversity of RAPD markers, pollen character and micropropagation of lily cultivars were studied. By combination of results in the present study with others that have been conducted in other previous research and analysis of diversity, breeding background of the above three strains , relationships among the cultivars, some conclusion expected to be drawed for the breeding of ornamentals lily in China.
Based on 15 random primers , RAPD polymorphism of 15 lilium cultivars were detected. Of 218 bands, 216 were polymorphic (99.08%) . The PPB, h and /of the Asiatic cultivars were 40.83%, 0.1654, 0.2415, respectively; the PPB, h and I of the Oriental cultivars were 42.20%, 0.1600, 0.2381, respectively; the PPB, h and /of the Longiflorum cultivars were 25.23%, 0.1072, 0.1551 respectively. The lily cultivars of the Asiatic hybrids had the highest genetic diversity; Longiflorum hybrids had the lowest genetic diversity.
Neis genetic distance among the cultivars showed that value of gernetic distance among the cultivars from the same strain were lower than that from different strains. The results revealed that cultivars from the same strain had much closer relationships; the cultivars from different strains had less close relationships. Among these three strains, gernetic distance between Asiatic cultivars and Oriental cultivars was the lowest, which was the highest between the Asiatic cultivars and Longiflorum cultivars. The Oriental strain and Asiatic strain had relatively closer relationship.
The pollen grains of some cultivars from the three hybrid strains were examined under
scanning electron microscope (SEM). The pollen character and gross morphology in Lilium were discussed: The pollen grains were generally elliptic and monocolpate(distal). Evidence from pollen morphology indicated that the pollen size, P/E value and exine sculpturing among species, cultivars were different in varying degrees, which could be used to distinguish cultivars. The main character of cultivars from the same strain are similar, which revealed that these cultivars had closer relationships. The exine sculpturing also revealed that there was more similarity between Asiatic Hybrids and Oriental Hybrids.
In present study, The difference in clonal propagation of the three strains cultivars were discussed through culture of scales. The induction proportion and the growth status of the buds and little bulbs, the proliferation of the buds and their growth status had direct correlation on genotype of lily. Cultivars from the same hybrid strain had more similarity .
The study showed the flower colour and shape were related with genetic diversity and relationships of lily cultivars. The Oriental and Asiatic hybrid cultivars were rich in flower colour and shape, they also had high genetic diversity. The Longiflorum hybrid cultivars was poor in flower colour and shape, they also had lower genetic diversity. In the same strain, cultivars that were same in flower colour were more familiar than others. The above results could be used to distinguish the relationship of lily cultivars from the same strains.
The main results are as follows:
1. The lily cultivars of the Asiatic hybrids strains had the highest genetic diversity; Longiflorum hybrid strains had the lowest gene
引文
1．邹喻苹，葛颂，王晓东．系统与进化植物学中的分子标记[M]．北京：科学出版社，2001，30～51．
2．黄敏玲，陈传林．透百合离体快速繁殖．植物生理学通讯，1993，29(6)．
3．杨成德，种康，敬兰花．百合鳞片不同部位的小鳞茎分化与调节因子调节研究．兰州大学学报(自然科学版)，1988，24(3)：95～99．
4．秦贺兰，游捷，高俊平．菊花18个品种的RAPD分析．园艺学报，2002，29(5)：488～490．
5．阮颖，周朴华，刘春林．九种李属植物的RAPD亲缘关系分析．园艺学报，2002，29(3)：218～223．
6．伍宁丰，李汝刚，伍晓明，朱莉等．中国甘蓝油菜遗传多样性的RAPD分子标记生物多样性．1997，5(4)：246～250．
7．黄富祥，王跃思．试论生物多样性保护理论与实践面临的困难及现实出路．生物多样性，2001，9(4)：399～406．
8．埃尔特曼 孢粉学手册[M]。科学出版社1978，
9．惠东威等．利用RAPD对大豆属植物系统学研究的初报．科学通报，1994，39(2)：175-178．
10．解新明，云锦凤．植物遗传多样性及其检测方法．中国草地，2000(6)：51-59．
11．孔令让等．粗山羊草随扩增多态性DNA研究．植物学报，1998，40(3)：223-227．
12．何川生，何兴金，葛颂等，李天飞，许凌云，许美玲，许介眉．烤烟品种资源的RAPD分析．植物学报，2001，43(6)：610-614．
13．马克平．试论生物多样性的概念．生物多样性，1993，1(1)：20-22．
14．钱迎倩，马克平．生物多样性研究的原理与方法[M]．北京：科学出版社，1993，99-113．
15．沈浩，刘登义．遗传多样性概述．生物学杂志，2001(6)：5-7．
16．施立明．遗传多样性及其保护．生物科学信息，1990，(2)：158-164．
17．桂枝，袁庆华，蔺海明等，RAPD标记技术及其在牧草中的应用。草业科学2001，18(2)：50～56．
18．曹家树，1997生物多样性的进化原理及保护对策．生物多样性5(30)：220～223．
19．赵祥云，陈新露珠，方海等．用RAPD标记评价百合品种间遗传关系。北京农学院学报，1995，10(2)：58～63。
20．陶铭，组织培养中畸形胚状体及超度含水态苗的研究．西北植物学报，2001，21(5)：1048-1058．
21．王开发、王宪曾，孢粉学概论[M]。北京大学出版社1983．
22．孙坤，马瑞君，杨永利．川蔓藻科植物的花粉形态研究．西北师范大学学报(自然科学版)，2002，38(1)：58～61．
23．刘杰，莫结胜，刘公社，齐冬梅，李芳芳．向日葵种质资源的随机扩增多态性DNA(RAPD)研究．植物学报，2001，43(2)：151-157．
24．孙坤，陈家宽，张志耘．水雍科植物的花粉形态研究．植物研究．2002，22(1)：33～38．
25．杨德奎，山东米口袋属花粉形态的研究．山东科学，2002，15(1)：17～19．
26．陈为明，宋为民．卷丹和青岛百合的组织培养及植株分化．植物生理学通讯．1982，2：35～36．
27．黄家福，陈振光．百合快速繁殖条件的优化．福建农业大学学报．1999，28(2)：152-156．
28．汤庚国，向其柏．樟科植物花粉形态研究．植物分类学报，1995，33(2)：161～170．
29．史肖白，蔡剑华．刺梨花粉形态观察．上海农业学报，1994，10(1)：88～92．
30．党荣理，姜彦成．新疆繁缕族漆姑草亚族植物花粉形态学研究．1995，12(2)：82～85．
31．党荣理，姜彦成．新疆裸果木亚科植物花粉形态学研究．1996，16(1)：61～64．
32．付桂香，顾志平，冯瑞芝，陈四保．国产葛属植物花粉形态的研究．热带亚热带植物学报，1996，4(2)：25～30．
33．施宗明，范眸天．芒果品种的花粉形态．云南植物研究，1998，20(1)：63～66．
34．张西丽，周厚高，周焱，宁云芬．百合品种间的数量分类研究．广西植物，2000，20(4)：325～328．
35．黄锦岭，刘宪章．开口箭属一新种及其核型和花粉形态．植物分类学报，1996，34(6)：592～596．
36．刘志铃，张友玉，李润堂，张映南．张家界地方梨花粉形态扫描电镜观察．湖南师范大学自然科学学报，2001，24(2)：76～79．
37．陈俊愉．主编，中国花卉品种分类学[M]．北京：中国林业出版社．
38．陈俊愉．1990，中国花经[M]．上海文化出版社．
39．龙雅宜等编著．1999，百合，球根花卉之王[M]．金盾出版社．
40．陈灵芝．中国的生物多样性——现状极其保护对策．北京：科学出版社，1993，1-9，99-113，210-212．
41．黄作喜，王祥宁，李克，等．百合鳞片扦插繁殖措施研究．天津农业科学，2001，7(4)：34～36．
42．邹喻苹，汪小全等．几种濒危植物及其近缘类群总DNA的提取与鉴定．植物学报，1994，36(7)：528～532．
43．邹喻苹，蔡美琳等．芍药属牡丹组的系统学研究——基于RAPD分析．植物分类学报，37(3)：220～227．
44．黄济民，赵晓艺．玫红百合为亲本育成百合种间杂种[J]．园艺学报，1990，2：153～156．
45．张西丽，周厚高，周焱．百合远缘杂交育种研究现状．广西农业生物科学，1999，18(2)：157-160．
46．马惠，郭扶兴，周俊彦，罗新谈．百合叶片愈伤组织的诱导和植株再生．植物生理学通讯，1992，28(4)：284～287．
47．杨柏云，罗丽萍，蔡奇英．切花百合的组织培养的研究。南昌大学学报(理科版)．2000，24(4)：323～326．
48．于晓英，吴铁明，倪沛，杨梦玲等．百合幼胚的离体培养和植株再生．湖南农业大学学报，2000，26(4)：296～288．
53．陈善娜，高建莉，郑志英．百合子房的组织培养．云南大学学报(自然科学版)1997，19(4)：374～375
49．周厚高，张西丽，王中仁，等．几个百合品种遗传亲缘关系的等位酶分析[J]．西南农业大学学报，1999，12(4)：92～95．
50．辛广，侯冬岩，刘琳琳，朱风妹．5种东方百合过氧化物酶同工酶的分析．鞍山师范学院学报，2003，5(6)：59～61．
51．高健，生物多样性的功能．盐城工学院学报．2001，14(4)：53～55．
52. Williams Kubelik A R, Livak K J. DNA polymorphisms amplified by arbitrary primers are useful as genetic makers. 1990, Nucl Acids Res 18:6231～6235.
53．张淑娟，刘与明．组织培养法快速繁殖新铁炮百合F1植株．植物生理学通讯，1998，34(5)：36．
54. Welsh J, Mcclelland M. Fingerprinting genomes using PCR with arbitrary primes. 1990, Nucl Acids Res, 18:7231～7218.
55. Okazaki K, YujiUmada. Interspecific Hybrids of Lilium longiflorum and
L.×formolongi with L.rubellum and L.japonicum through Embryo Culture. J Japan SocHort Sci, 1992, 60(4):997-1002.
56. JIA Jian-Hang, WANG Ping, JIN De-Min, QU Xue-Ping, WANG Qian, LI Chuan-You, WENG Man-Li, WANG Bing. The Application of RAPD Marker in Diversity Detection and Variety Identification Of Porphyra. Acta Botanica Sinica, 2000, 42(4):403-407.
57. DU Dao-Lin, SU Jie, ZHOU Peng, ZHENG Xue-Qin, HUANG Big-Zhi, LI Feng-Nian. RAPD Analysis of 33 Varieties of Banana. Acta Botanica Sinica, 2001, 43(10):1036-1042.
58. LUO Su-Lan, HE Pu-Chao, ZHENG Xue-Qin, ZHOU Peng, Genetic Diversity in Wild Grapes Native to China Based on Randomly Amplified Polymorphic DNA (RAPD) Analysis. Acta Botanica Sinica, 2001, 43(2): 158-163.
59. C. Wawrosch·P.R. Malla·B. Kopp. Clonal propagation of Lilium nepalenseD.Don, a threatened medicinal plant of Nepal. Plant Cell Reports (2001)20:285-288.
60. A.-M. Arzate-Fern(?)ndez·T. Nakazaki·Y. Okumoto, T. Tanisaka. Efficient callus induction and plant regeneration from filaments with anther in lily (Lilium longiflorum Thunb.) Plant Cell Reports (1997) 16: 836-840.
61. A. Tribulato·P. C. Remotti·H. J. M. L(?)ffier J. M. van Tuyl. Somatic embryogenesis and plant regeneration in Lilium longiflorum Thunb. Plant Cell Reports (1997) 17: 113-118.
62. Duong Tan Nhut, Bui Van Le, Michio Tanaka, K. Tran Thanh Van. Shoot induction and plant regeneration from receptacle tissues of Lilium longiflorum Scientia Horticulturae 87 (2001) 131-138.
63. Mitsugu Horita Hitomi Morohashi Fuminori Komai. Regeneration of flowering plants from difficile lily protoplasts by means of a nurse culture. Planta (2002) 215: 880-884.
64．马小军，汪小全，徐昭玺，肖培根，洪德元．人生不同栽培群体遗传关系的RAPD分析．植物学报，2000，42(6)：587-590．
65. Masumi Yamagishi, Hiromi Abe, Michiharu Nakano, Akira Nakatsuka. PCR-based molecular markers in Asiatic hybrid lily. Scientia Horticulturae (2002)96: 225-234
66. Haw, S＞G. The lilies of China: the genera Lilium Cardiocrinum, Nomochairs and Nothlirion, B.T.Batsford Ltd, London. 1986
67. Schall, B. A. et al. Comparison of methods for assessing genetic variation in plant conservation biology, In Falk D. A. and K. E. Holsinger(eds.) Genetics and Conservation of Rare Plants, New York: Oxford University Press. 1991, 123-134.
68. Gareth Lloyd-Jone, David W.F. Hunter. Characterization of fluoranthene-and pyrene-degrading Mycobacterium-like strains by RAPD and SSU sequencing. FEMS Microbiology Letters 1997,153 51-56.
69. Hui-Lee Wong, Hock-Hin Yeoh,Saw-Hoon Lim and Kwok-Chang Lionel Looi. Desing of primers for RAPD analysis of cassava manihot esculenta. Phytochemistrv, 1997,46 (5):805-810.
70. Pooja Jain, Zafar K. Khan,, Esha Bhattacharya, Shirish A. Ranade Variation in random amplified polymorphie DNA (RAPD)profiles specific to fluconazole-resistant and-sensitive strains of Candida albicans. Diagnostic Microbiology and Infectious Disease 41 (2001) 113-119.
71. Shi Liming, Genetic diversity and its preservation. Chinese Biodiversity, 1993, 1(1):23～31.
72. Singh R K, Genetic resource and the role of international collaboration in rice breeding. Genome, 1999.42:635～641.
73. Abo-elwafa A, Murai K, Shimada T. 1995. Intra-and Inter-specific variation in Lens revealed by RAPD marker. Theor Appl Genet, 90: 335～340.
74. Buso G S C, Rangel P H, Ferreira M E. 1998. Analysis of genetic variability of South American wild rice population (Oryza glumaepatula) with isozames and RAPD marker. Mol Ecol., 7:107～117.
75．曹孜义，刘国民．实用植物组织培养技术教程．甘肃科学技术出版社，1998．
76. Koller B, Lehmann A, MeDermolt J M. 1993. Identification of apple cultivars using RAPD marker. Theor Appl Genet. 85:901～904.
77. Torres A M, Weeden N F, Martin A. 1993. Linkage among isozyme, RFLP and RAPD marker in vicia faba. Theor Appl Genet..85:937～945.
78. North C, Will A B. Interspecific hybrids of Lilium lankongense Franchel produced by Embryo-cuiture. Euphytica, 1969, 18:430～4342.
79. Ronald W G, Ascher P D. Lilium'Black Beauty'-A Potential Hibrid in Lilium. uphytica, 1976, 25: 285～2913.
80. Asano Y, Hiroshi Myodo. Studies on Crosses between Distantly Related Species of Lilies. For the Intrastylar Poll ination Techique. J Japan Soc Hort Sci, 1997, 46(1):59～654.
81. Tuyl J M, Van Dien M R Application of in virtro Pollination, Ovary Culture and Embryo Rescue for Overcoming Incongriuity Barriers in Interspesific Lilium Crosses. Plant Science, 1991, 74:115～1265.
82. Matso, E, et al. Cultural Practices Influencing Premature Daughter Leaf and/or Shoot Emergence in Scale-Propagated Easter Lily. Hortscience, 1982, 17(2): 196～198.
83. Matso, E, et al. Factors Influencing Stem Root Emergence during Scale Propagation in the Easter Lily. Hortscience, 1983, 18(1): 78～79.
84. Matso, E, et al. Effect of Root Removal and Bulblet Size on Leaf Emergence from Scale Bulblets of Easter Lily. Hortscience, 1986, 21(4): 1033～1034.
85. Matso, E, and J. M. Erly Scale Propagation Results in Forcible Bulbs of Easter Lily. Hortscience, 1986, 21(4): 1003～1007.
86．郭志刚、张伟，花卉生产技术原理及其应用丛书，球根类[M]．中国林业大学出版社，清华大学出版社．
87. Yumi Obata, Yoshiji Niimi, Masaru Nakano, Keiichi Okazaki, Ichiro Miyajima. Interspecific hybrids between Lilium nobilissimum and L. regale produced via ovules-with-placental-tissue culture. Scientia Horticulturae 84 (2000) 191～204.
88. Amaury-M, Arzate-Fernandez, Tetsuya Nakazaki, Hirotada Yamagata, Takatoshi Tanisaka. Production of double-haploid plants from Lilium longiflorum Thunb. Anther culture. Plant Science 123(1997) 179～187.
89. D. T. Nhut. Micropropagation of lily (Lilium longiflorum) via in vitro stem nodeand pseudo-bulblet culture. Plant Cell Reports. (1998) 17:913～916.
90. Fernandez A M,Nakazaki T. Development of diploid and triploid interspecific hybrids between Lilium longiflorum and L. Concolor by ovary slice culture. Plant Breeding, 1996,115:167～171.
91. Li Q M, Xu Z F, He T H. The study on the conservation genetics of endangered Vatica guangxiensis (Dipterocarpanceae), Acta Botanica Sinica 2002, 44(2): 246～249.
2．黄敏玲，陈传林．透百合离体快速繁殖．植物生理学通讯，1993，29(6)．
3．杨成德，种康，敬兰花．百合鳞片不同部位的小鳞茎分化与调节因子调节研究．兰州大学学报(自然科学版)，1988，24(3)：95～99．
4．秦贺兰，游捷，高俊平．菊花18个品种的RAPD分析．园艺学报，2002，29(5)：488～490．
5．阮颖，周朴华，刘春林．九种李属植物的RAPD亲缘关系分析．园艺学报，2002，29(3)：218～223．
6．伍宁丰，李汝刚，伍晓明，朱莉等．中国甘蓝油菜遗传多样性的RAPD分子标记生物多样性．1997，5(4)：246～250．
7．黄富祥，王跃思．试论生物多样性保护理论与实践面临的困难及现实出路．生物多样性，2001，9(4)：399～406．
8．埃尔特曼 孢粉学手册[M]。科学出版社1978，
9．惠东威等．利用RAPD对大豆属植物系统学研究的初报．科学通报，1994，39(2)：175-178．
10．解新明，云锦凤．植物遗传多样性及其检测方法．中国草地，2000(6)：51-59．
11．孔令让等．粗山羊草随扩增多态性DNA研究．植物学报，1998，40(3)：223-227．
12．何川生，何兴金，葛颂等，李天飞，许凌云，许美玲，许介眉．烤烟品种资源的RAPD分析．植物学报，2001，43(6)：610-614．
13．马克平．试论生物多样性的概念．生物多样性，1993，1(1)：20-22．
14．钱迎倩，马克平．生物多样性研究的原理与方法[M]．北京：科学出版社，1993，99-113．
15．沈浩，刘登义．遗传多样性概述．生物学杂志，2001(6)：5-7．
16．施立明．遗传多样性及其保护．生物科学信息，1990，(2)：158-164．
17．桂枝，袁庆华，蔺海明等，RAPD标记技术及其在牧草中的应用。草业科学2001，18(2)：50～56．
18．曹家树，1997生物多样性的进化原理及保护对策．生物多样性5(30)：220～223．
19．赵祥云，陈新露珠，方海等．用RAPD标记评价百合品种间遗传关系。北京农学院学报，1995，10(2)：58～63。
20．陶铭，组织培养中畸形胚状体及超度含水态苗的研究．西北植物学报，2001，21(5)：1048-1058．
21．王开发、王宪曾，孢粉学概论[M]。北京大学出版社1983．
22．孙坤，马瑞君，杨永利．川蔓藻科植物的花粉形态研究．西北师范大学学报(自然科学版)，2002，38(1)：58～61．
23．刘杰，莫结胜，刘公社，齐冬梅，李芳芳．向日葵种质资源的随机扩增多态性DNA(RAPD)研究．植物学报，2001，43(2)：151-157．
24．孙坤，陈家宽，张志耘．水雍科植物的花粉形态研究．植物研究．2002，22(1)：33～38．
25．杨德奎，山东米口袋属花粉形态的研究．山东科学，2002，15(1)：17～19．
26．陈为明，宋为民．卷丹和青岛百合的组织培养及植株分化．植物生理学通讯．1982，2：35～36．
27．黄家福，陈振光．百合快速繁殖条件的优化．福建农业大学学报．1999，28(2)：152-156．
28．汤庚国，向其柏．樟科植物花粉形态研究．植物分类学报，1995，33(2)：161～170．
29．史肖白，蔡剑华．刺梨花粉形态观察．上海农业学报，1994，10(1)：88～92．
30．党荣理，姜彦成．新疆繁缕族漆姑草亚族植物花粉形态学研究．1995，12(2)：82～85．
31．党荣理，姜彦成．新疆裸果木亚科植物花粉形态学研究．1996，16(1)：61～64．
32．付桂香，顾志平，冯瑞芝，陈四保．国产葛属植物花粉形态的研究．热带亚热带植物学报，1996，4(2)：25～30．
33．施宗明，范眸天．芒果品种的花粉形态．云南植物研究，1998，20(1)：63～66．
34．张西丽，周厚高，周焱，宁云芬．百合品种间的数量分类研究．广西植物，2000，20(4)：325～328．
35．黄锦岭，刘宪章．开口箭属一新种及其核型和花粉形态．植物分类学报，1996，34(6)：592～596．
36．刘志铃，张友玉，李润堂，张映南．张家界地方梨花粉形态扫描电镜观察．湖南师范大学自然科学学报，2001，24(2)：76～79．
37．陈俊愉．主编，中国花卉品种分类学[M]．北京：中国林业出版社．
38．陈俊愉．1990，中国花经[M]．上海文化出版社．
39．龙雅宜等编著．1999，百合，球根花卉之王[M]．金盾出版社．
40．陈灵芝．中国的生物多样性——现状极其保护对策．北京：科学出版社，1993，1-9，99-113，210-212．
41．黄作喜，王祥宁，李克，等．百合鳞片扦插繁殖措施研究．天津农业科学，2001，7(4)：34～36．
42．邹喻苹，汪小全等．几种濒危植物及其近缘类群总DNA的提取与鉴定．植物学报，1994，36(7)：528～532．
43．邹喻苹，蔡美琳等．芍药属牡丹组的系统学研究——基于RAPD分析．植物分类学报，37(3)：220～227．
44．黄济民，赵晓艺．玫红百合为亲本育成百合种间杂种[J]．园艺学报，1990，2：153～156．
45．张西丽，周厚高，周焱．百合远缘杂交育种研究现状．广西农业生物科学，1999，18(2)：157-160．
46．马惠，郭扶兴，周俊彦，罗新谈．百合叶片愈伤组织的诱导和植株再生．植物生理学通讯，1992，28(4)：284～287．
47．杨柏云，罗丽萍，蔡奇英．切花百合的组织培养的研究。南昌大学学报(理科版)．2000，24(4)：323～326．
48．于晓英，吴铁明，倪沛，杨梦玲等．百合幼胚的离体培养和植株再生．湖南农业大学学报，2000，26(4)：296～288．
53．陈善娜，高建莉，郑志英．百合子房的组织培养．云南大学学报(自然科学版)1997，19(4)：374～375
49．周厚高，张西丽，王中仁，等．几个百合品种遗传亲缘关系的等位酶分析[J]．西南农业大学学报，1999，12(4)：92～95．
50．辛广，侯冬岩，刘琳琳，朱风妹．5种东方百合过氧化物酶同工酶的分析．鞍山师范学院学报，2003，5(6)：59～61．
51．高健，生物多样性的功能．盐城工学院学报．2001，14(4)：53～55．
52. Williams Kubelik A R, Livak K J. DNA polymorphisms amplified by arbitrary primers are useful as genetic makers. 1990, Nucl Acids Res 18:6231～6235.
53．张淑娟，刘与明．组织培养法快速繁殖新铁炮百合F1植株．植物生理学通讯，1998，34(5)：36．
54. Welsh J, Mcclelland M. Fingerprinting genomes using PCR with arbitrary primes. 1990, Nucl Acids Res, 18:7231～7218.
55. Okazaki K, YujiUmada. Interspecific Hybrids of Lilium longiflorum and
L.×formolongi with L.rubellum and L.japonicum through Embryo Culture. J Japan SocHort Sci, 1992, 60(4):997-1002.
56. JIA Jian-Hang, WANG Ping, JIN De-Min, QU Xue-Ping, WANG Qian, LI Chuan-You, WENG Man-Li, WANG Bing. The Application of RAPD Marker in Diversity Detection and Variety Identification Of Porphyra. Acta Botanica Sinica, 2000, 42(4):403-407.
57. DU Dao-Lin, SU Jie, ZHOU Peng, ZHENG Xue-Qin, HUANG Big-Zhi, LI Feng-Nian. RAPD Analysis of 33 Varieties of Banana. Acta Botanica Sinica, 2001, 43(10):1036-1042.
58. LUO Su-Lan, HE Pu-Chao, ZHENG Xue-Qin, ZHOU Peng, Genetic Diversity in Wild Grapes Native to China Based on Randomly Amplified Polymorphic DNA (RAPD) Analysis. Acta Botanica Sinica, 2001, 43(2): 158-163.
59. C. Wawrosch·P.R. Malla·B. Kopp. Clonal propagation of Lilium nepalenseD.Don, a threatened medicinal plant of Nepal. Plant Cell Reports (2001)20:285-288.
60. A.-M. Arzate-Fern(?)ndez·T. Nakazaki·Y. Okumoto, T. Tanisaka. Efficient callus induction and plant regeneration from filaments with anther in lily (Lilium longiflorum Thunb.) Plant Cell Reports (1997) 16: 836-840.
61. A. Tribulato·P. C. Remotti·H. J. M. L(?)ffier J. M. van Tuyl. Somatic embryogenesis and plant regeneration in Lilium longiflorum Thunb. Plant Cell Reports (1997) 17: 113-118.
62. Duong Tan Nhut, Bui Van Le, Michio Tanaka, K. Tran Thanh Van. Shoot induction and plant regeneration from receptacle tissues of Lilium longiflorum Scientia Horticulturae 87 (2001) 131-138.
63. Mitsugu Horita Hitomi Morohashi Fuminori Komai. Regeneration of flowering plants from difficile lily protoplasts by means of a nurse culture. Planta (2002) 215: 880-884.
64．马小军，汪小全，徐昭玺，肖培根，洪德元．人生不同栽培群体遗传关系的RAPD分析．植物学报，2000，42(6)：587-590．
65. Masumi Yamagishi, Hiromi Abe, Michiharu Nakano, Akira Nakatsuka. PCR-based molecular markers in Asiatic hybrid lily. Scientia Horticulturae (2002)96: 225-234
66. Haw, S＞G. The lilies of China: the genera Lilium Cardiocrinum, Nomochairs and Nothlirion, B.T.Batsford Ltd, London. 1986
67. Schall, B. A. et al. Comparison of methods for assessing genetic variation in plant conservation biology, In Falk D. A. and K. E. Holsinger(eds.) Genetics and Conservation of Rare Plants, New York: Oxford University Press. 1991, 123-134.
68. Gareth Lloyd-Jone, David W.F. Hunter. Characterization of fluoranthene-and pyrene-degrading Mycobacterium-like strains by RAPD and SSU sequencing. FEMS Microbiology Letters 1997,153 51-56.
69. Hui-Lee Wong, Hock-Hin Yeoh,Saw-Hoon Lim and Kwok-Chang Lionel Looi. Desing of primers for RAPD analysis of cassava manihot esculenta. Phytochemistrv, 1997,46 (5):805-810.
70. Pooja Jain, Zafar K. Khan,, Esha Bhattacharya, Shirish A. Ranade Variation in random amplified polymorphie DNA (RAPD)profiles specific to fluconazole-resistant and-sensitive strains of Candida albicans. Diagnostic Microbiology and Infectious Disease 41 (2001) 113-119.
71. Shi Liming, Genetic diversity and its preservation. Chinese Biodiversity, 1993, 1(1):23～31.
72. Singh R K, Genetic resource and the role of international collaboration in rice breeding. Genome, 1999.42:635～641.
73. Abo-elwafa A, Murai K, Shimada T. 1995. Intra-and Inter-specific variation in Lens revealed by RAPD marker. Theor Appl Genet, 90: 335～340.
74. Buso G S C, Rangel P H, Ferreira M E. 1998. Analysis of genetic variability of South American wild rice population (Oryza glumaepatula) with isozames and RAPD marker. Mol Ecol., 7:107～117.
75．曹孜义，刘国民．实用植物组织培养技术教程．甘肃科学技术出版社，1998．
76. Koller B, Lehmann A, MeDermolt J M. 1993. Identification of apple cultivars using RAPD marker. Theor Appl Genet. 85:901～904.
77. Torres A M, Weeden N F, Martin A. 1993. Linkage among isozyme, RFLP and RAPD marker in vicia faba. Theor Appl Genet..85:937～945.
78. North C, Will A B. Interspecific hybrids of Lilium lankongense Franchel produced by Embryo-cuiture. Euphytica, 1969, 18:430～4342.
79. Ronald W G, Ascher P D. Lilium'Black Beauty'-A Potential Hibrid in Lilium. uphytica, 1976, 25: 285～2913.
80. Asano Y, Hiroshi Myodo. Studies on Crosses between Distantly Related Species of Lilies. For the Intrastylar Poll ination Techique. J Japan Soc Hort Sci, 1997, 46(1):59～654.
81. Tuyl J M, Van Dien M R Application of in virtro Pollination, Ovary Culture and Embryo Rescue for Overcoming Incongriuity Barriers in Interspesific Lilium Crosses. Plant Science, 1991, 74:115～1265.
82. Matso, E, et al. Cultural Practices Influencing Premature Daughter Leaf and/or Shoot Emergence in Scale-Propagated Easter Lily. Hortscience, 1982, 17(2): 196～198.
83. Matso, E, et al. Factors Influencing Stem Root Emergence during Scale Propagation in the Easter Lily. Hortscience, 1983, 18(1): 78～79.
84. Matso, E, et al. Effect of Root Removal and Bulblet Size on Leaf Emergence from Scale Bulblets of Easter Lily. Hortscience, 1986, 21(4): 1033～1034.
85. Matso, E, and J. M. Erly Scale Propagation Results in Forcible Bulbs of Easter Lily. Hortscience, 1986, 21(4): 1003～1007.
86．郭志刚、张伟，花卉生产技术原理及其应用丛书，球根类[M]．中国林业大学出版社，清华大学出版社．
87. Yumi Obata, Yoshiji Niimi, Masaru Nakano, Keiichi Okazaki, Ichiro Miyajima. Interspecific hybrids between Lilium nobilissimum and L. regale produced via ovules-with-placental-tissue culture. Scientia Horticulturae 84 (2000) 191～204.
88. Amaury-M, Arzate-Fernandez, Tetsuya Nakazaki, Hirotada Yamagata, Takatoshi Tanisaka. Production of double-haploid plants from Lilium longiflorum Thunb. Anther culture. Plant Science 123(1997) 179～187.
89. D. T. Nhut. Micropropagation of lily (Lilium longiflorum) via in vitro stem nodeand pseudo-bulblet culture. Plant Cell Reports. (1998) 17:913～916.
90. Fernandez A M,Nakazaki T. Development of diploid and triploid interspecific hybrids between Lilium longiflorum and L. Concolor by ovary slice culture. Plant Breeding, 1996,115:167～171.
91. Li Q M, Xu Z F, He T H. The study on the conservation genetics of endangered Vatica guangxiensis (Dipterocarpanceae), Acta Botanica Sinica 2002, 44(2): 246～249.