草原龙胆生物学特性及遗传研究
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摘要
一、本研究在探讨草原龙胆生物学特性过程中进行了如下研究：采取不同的播种时间比较植株生长的差异；利用光学显微镜观察营养器官及繁殖器官的结构；利用石蜡切片法研究了花芽分化过程以及大孢子的发生、胚和胚乳的发育过程，并且根据花蕾生长数据分析了花的生长趋势；利用荧光显微法观察了在柱头上的花粉萌发情况；采用不同的光照、水分、温度等条件观察了种子的萌发与植物的生长特性。
对生物学特性研究的主要结果如下：北方地区播种时间选择在11月到12月份比较适宜；主枝与侧枝花蕾直径的发育进程呈现出S型的变化趋势，主枝与侧枝花蕾直径与花蕾长度发育的对应关系都经过较快的前期生长及平稳的后期生长过程；单瓣花和重瓣花花芽分化过程均可分为萼片原基分化期、花瓣原基分化期、雄蕊原基分化期和雌蕊原基分化期5个时期，其重瓣类型属于花冠重复类型；雌配子体发育属于蓼型，成熟种子的胚大部分发育到心形胚时期；种子处理采用48小时的浸种，提高萌发率最高；在有光照的条件下，20～25℃是种子萌发的适宜温度，萌发率和萌发势均较高，且幼苗后期生长健壮。
二、本研究在探讨草原龙胆的花色及花型遗传特性研究结果如下：
1 根据对花瓣中所含的色素种类进行测定的吸收峰可以初步确定，紫色所含色素可能由花青素构成；白色花瓣中可能存在类黄酮；红色花有两个吸收峰，可能由类黄酮和花青素两种色素，绿色花可能含有以叶绿素为主的色素类群。
2 利用常规的染色体制片方法，选择根尖以及长度为7mm-8mm的花苞中的花药进行有丝分裂和减数分裂过程的观察，并且观察到草原龙胆的染色体为72条。
3 利用全轮配模型初步探讨了草原龙胆的花色遗传特性，从后代表现可以初步看出，同一色系不同品系间杂交，后代表现亲本的花色；紫色具有明显的显性性状；其他花色之间进行杂交，后代颜色趋于父母本的中间色偏颜色较深的亲本。
4 根据现有草原龙胆品系中花型具有明显特征的四个品系(钟状、漏斗状、杯状和平状)，建立三步花型比较模型，逐步进行筛选，以确定未知种类的花型类型。
5 通过杂交的手段初步研究了四种花型之间的遗传特性，根据杂交结果可以初步看出，无论正反交，钟状与漏斗状品系的杂交后代都表现漏斗状；钟状与平状杂交后代花型表现为钟状，杯状与平状杂交后代花型表现为杯状。
Firstly, studies were made on biological characteristics of Lisianthus: By using the diffrerent time of planting seeds, the growth differences were compared. Vegetative organs and reproductive organs were observed with a microscope. Floral development, the megasporogenesis, and development of embryos and endosperms were observed microscopically after making FAA-fixed paraffin sections. The growth pattern of a flower was analyzing from data of perianth growth. Pollen germination was observed with a fluorescence microscope. Germination characteristics of seeds were investigated under controlled conditions of illumination, moisture and temperature.
The results that studies were made on biological characteristics are as followed. In northern regions where it is very cold in winter, the best sowing time of seeds ranged from November to December in a greenhouse. The growth in diameter of a perianth on primary and second branches showed a pattern of S shape. Growth in diameter and length of a perianth was rapid during a first half of a growth period but slower during a second half. Floral development in both types of flowers, i.e., single and double flowers, was divided into five stages in view of the beginning of following events: Appearance of (Stage 1) flower buttress and the initiation of development, (Stage 2) sepal primordia, (Stage 3) petal primordia, (Stage 4) stamen primordia, and (Stage 5) gynoecium primordia. In double flowers, their type of flower shape were corolla repeated The development of female gametophyte was of Polygonum type, and mature seeds only developed during a late period of globular embryo stage. Time of soaking seeds required f
or best germination was 48 hours. A high rate of seed germination was shown at 20 to 25 C in light conditions and healthy seedlings were produced.
Secondly, studies were made on genetic characteristics of flower color and shape.
1. Pigments in petals were examined by peak values of absorbance measured with a UV photometer. Peak absorbance of pigments from purple petals and pink petals were suggested the presence of cyanine. Peak absorbance of pigments from white petals was suggested the presence of flavonoids. Pigments from red petals showed two peaks, suggesting a mixture of flavonoids and cyanine. Pigments from green petals was caused by chlorophylls.
2. The process of mitosis and meiosis in root tips and anthers in a perianth, whose length was 7 to 8mm, were observed by using a method to make slices for chromosome observation. The results showed that number of chromosome was 72.
3. Inheritance characteristics of flower color was carried out using full-turns mating model. From the expression in individuals of progenies, it was suggested that crosses between different
lines with the same flower color produced progenies having the same color with that of parents. Purple color was obviously a dominant characteristic. Crosses between different lines with different flower color produced progenies having color between that of parents, but the color intensity tended to resemble that of a parent showing a darker color.
4. Using flower shapes of four varieties (clock form, funnel form, cup form and flat form) that had distinct characteristic, three stepwise comparison models were proposed. These models could help to show flower shape of unknown varieties.
5. The genetic characteristics of four shapes were determined tentatively through the results of hybridization. Crosses between clock form and funnel form produced progenies with funnel form, even in a reciprocal crossing. Crosses between clock form and flat form produced progenies with clock form. Crosses between cup form and flat form produced progenies with cup form.
对生物学特性研究的主要结果如下：北方地区播种时间选择在11月到12月份比较适宜；主枝与侧枝花蕾直径的发育进程呈现出S型的变化趋势，主枝与侧枝花蕾直径与花蕾长度发育的对应关系都经过较快的前期生长及平稳的后期生长过程；单瓣花和重瓣花花芽分化过程均可分为萼片原基分化期、花瓣原基分化期、雄蕊原基分化期和雌蕊原基分化期5个时期，其重瓣类型属于花冠重复类型；雌配子体发育属于蓼型，成熟种子的胚大部分发育到心形胚时期；种子处理采用48小时的浸种，提高萌发率最高；在有光照的条件下，20～25℃是种子萌发的适宜温度，萌发率和萌发势均较高，且幼苗后期生长健壮。
二、本研究在探讨草原龙胆的花色及花型遗传特性研究结果如下：
1 根据对花瓣中所含的色素种类进行测定的吸收峰可以初步确定，紫色所含色素可能由花青素构成；白色花瓣中可能存在类黄酮；红色花有两个吸收峰，可能由类黄酮和花青素两种色素，绿色花可能含有以叶绿素为主的色素类群。
2 利用常规的染色体制片方法，选择根尖以及长度为7mm-8mm的花苞中的花药进行有丝分裂和减数分裂过程的观察，并且观察到草原龙胆的染色体为72条。
3 利用全轮配模型初步探讨了草原龙胆的花色遗传特性，从后代表现可以初步看出，同一色系不同品系间杂交，后代表现亲本的花色；紫色具有明显的显性性状；其他花色之间进行杂交，后代颜色趋于父母本的中间色偏颜色较深的亲本。
4 根据现有草原龙胆品系中花型具有明显特征的四个品系(钟状、漏斗状、杯状和平状)，建立三步花型比较模型，逐步进行筛选，以确定未知种类的花型类型。
5 通过杂交的手段初步研究了四种花型之间的遗传特性，根据杂交结果可以初步看出，无论正反交，钟状与漏斗状品系的杂交后代都表现漏斗状；钟状与平状杂交后代花型表现为钟状，杯状与平状杂交后代花型表现为杯状。
Firstly, studies were made on biological characteristics of Lisianthus: By using the diffrerent time of planting seeds, the growth differences were compared. Vegetative organs and reproductive organs were observed with a microscope. Floral development, the megasporogenesis, and development of embryos and endosperms were observed microscopically after making FAA-fixed paraffin sections. The growth pattern of a flower was analyzing from data of perianth growth. Pollen germination was observed with a fluorescence microscope. Germination characteristics of seeds were investigated under controlled conditions of illumination, moisture and temperature.
The results that studies were made on biological characteristics are as followed. In northern regions where it is very cold in winter, the best sowing time of seeds ranged from November to December in a greenhouse. The growth in diameter of a perianth on primary and second branches showed a pattern of S shape. Growth in diameter and length of a perianth was rapid during a first half of a growth period but slower during a second half. Floral development in both types of flowers, i.e., single and double flowers, was divided into five stages in view of the beginning of following events: Appearance of (Stage 1) flower buttress and the initiation of development, (Stage 2) sepal primordia, (Stage 3) petal primordia, (Stage 4) stamen primordia, and (Stage 5) gynoecium primordia. In double flowers, their type of flower shape were corolla repeated The development of female gametophyte was of Polygonum type, and mature seeds only developed during a late period of globular embryo stage. Time of soaking seeds required f
or best germination was 48 hours. A high rate of seed germination was shown at 20 to 25 C in light conditions and healthy seedlings were produced.
Secondly, studies were made on genetic characteristics of flower color and shape.
1. Pigments in petals were examined by peak values of absorbance measured with a UV photometer. Peak absorbance of pigments from purple petals and pink petals were suggested the presence of cyanine. Peak absorbance of pigments from white petals was suggested the presence of flavonoids. Pigments from red petals showed two peaks, suggesting a mixture of flavonoids and cyanine. Pigments from green petals was caused by chlorophylls.
2. The process of mitosis and meiosis in root tips and anthers in a perianth, whose length was 7 to 8mm, were observed by using a method to make slices for chromosome observation. The results showed that number of chromosome was 72.
3. Inheritance characteristics of flower color was carried out using full-turns mating model. From the expression in individuals of progenies, it was suggested that crosses between different
lines with the same flower color produced progenies having the same color with that of parents. Purple color was obviously a dominant characteristic. Crosses between different lines with different flower color produced progenies having color between that of parents, but the color intensity tended to resemble that of a parent showing a darker color.
4. Using flower shapes of four varieties (clock form, funnel form, cup form and flat form) that had distinct characteristic, three stepwise comparison models were proposed. These models could help to show flower shape of unknown varieties.
5. The genetic characteristics of four shapes were determined tentatively through the results of hybridization. Crosses between clock form and funnel form produced progenies with funnel form, even in a reciprocal crossing. Crosses between clock form and flat form produced progenies with clock form. Crosses between cup form and flat form produced progenies with cup form.
引文
1 [日]安田齐，1989．花色之谜．北京：中国林业出版社
2 [日]大川清，2000．花专科育种栽培Eustoma．诚文堂新光社，10～14
3 包满珠，1991．植物花青素基因的克隆及应用．园艺学报，24(3)：279～284
4 程金水，2001．园林植物遗传育种学．北京：中国林业出版社
5 方开泰，全辉，陈庆云，1988．实用回归分析．北京：科学出版社，114～116
6 傅荣昭，马江生，曹光诚，等，1995．观赏植物色香形基因工程研究进展．园艺学报，22(4)：381～385
7 高信曾，1986．植物学试验指导(形态解剖部分)．北京：高等教育出版社，58～179
8 郝立勤，顾金凤，2002．云南花卉产业发展对策研究．云南科技管理，4：12～15
9 何凤仙，2000．植物学试验．北京：高等教育出版社，41
10 何廷农，陈世龙，2000．条纹龙胆的胚胎学研究．西北植物学报，20(6)：960～967
11 何廷农，刘建全，1999．线叶龙胆的胚胎学研究．西北植物学报，19(2)：234～240
12 何奕昆，1995．雁来红叶色转变与超微结构及色素含量的关系．四川师范学院学报，16(3)：86～90
13 黄素梅，蓝福生，姚军，等，2000．我国花卉产业现状及其发展前景探讨．广西植物，20(3)：277～283
14 黄胤怡，沈明山，李鹏，等，2002．蓝色花基因工程．植物生理学通讯，38(2)：203～206
15 及华，温春秀，高延厅，1996．洋桔梗的离体快速繁殖，32(6)：431～432
16 及华，1999．日本的花卉业及生产经营．河北科技．2：38～41
17 金波，东惠茹，1994．一品红花色的探讨．园艺学报，21(1)：87～90
18 李嘉钰．1999，中国牡丹与芍药．北京：中国林业出版社，103～116
19 李懋学，张敩方，1991．植物染色体研究技术．哈尔滨：东北林业大学出版社，1～58
20 李正理，张新英，1987．植物解剖学．北京：高等教育出版社，166～293
21 林功涛，李凤宜，2000．北京小菊品系选育及遗传规律探讨．中国花卉科技二十年，北京：科学技术出版社，434～444
22 刘穆，2000．种子植物形态解剖学导论．北京：科学出版社，98～319
23 刘青林，陈俊愉，1998．观赏植物花器官主要观赏性状的遗传与改良．园艺学报，25(1)：81～86
24 潘会堂，张启翔，2000．花卉种质资源与遗传育种研究进展．北京林业大学学报，22(1)：81～86
25 潘家驹，1992．作物育种学总论．北京：中国农业大学出版社，50～56
26 庞志中，2000．花色研究概况．北京农业科学，18(5)：34～387
27 宋晓荭，文绍金，韩文婷，等，2003．借鉴经验，强化特色，加快结构调整步伐．青海农技推广，2：61～62
28 孙振雷，1999．观赏植物育种学．北京：民族出版社，45～49
29 孙自然，游捷，2000．我国菊花新品系选育进展．中国花卉科技二十年，北京：科学技术出版社，414～423
30 唐道城，张志英，赵梁军，2001．一串红花芽形态分化进程．园艺学报，28(4)：367～369
31 陶嘉龄，郑光华，1991．种子活力．北京：科学出版社，108～110
32 王瑗玲，1998a．洋桔梗的栽培(一)．花木盆景，25(1)：26
33 王瑷玲，1998b．洋桔梗的栽培(二)．花木盆景，26(2)：22～23
34 王钦丽，卢龙斗，吴小琴，2002．花粉的保存及其生活力测定．植物学通报，19(3)：365～373
35 王少平，朱爱民，2002．浸种对波斯菊种子发芽的影响．种子，108(2)：39～40
36 王仲礼，刘林德，田伟国，1998．短柄五加大、小孢子发生和雌雄配子体发育的研究．植物研究，18(2)：177～183
37 王仲礼，刘林德，田伟国，1999．短柄五加胚和胚乳发育的研究．植物研究，19(1)：41～46
38 吴昌陆，胡南珍，1995．蜡梅花部形态和开花习性研究．园艺学报，22(3)：277～282
39 吴人坚，1987．植物学试验方法．上海：上海科学技术出版社，52～77
40 夏宜平，2001．鲜切花培育技术．上海：上海科学技术出版社，85～86
41 徐克学，1999．生物数学．北京：科学出版社，40～49
42 徐振华，郭伟珍，王鑫，1997．洋桔梗的种子育苗技术．河北林业科技，4：43
43 薛淮，刘敏，张纯花，等，2002，花卉分子育种研究进展．生物工程进展，22(2)：80～83
44 杨继，2000．植物生物学试验．北京：高等教育出版社，施普林格出版社，67～117
45 杨霞，徐康，1997．洋桔梗的组织培养及快速繁殖．植物生理学通讯，33(6)：435～438
46 杨永刚，周根余，2001．洋桔梗叶片体外再生系统的激素调控．上海师范大学学报(自然科学版)，3(1)：99～100
47 余地求，李宝健，1997．花色素苷生物合成的遗传和发育调控．植物生理学通讯，33(1)：71～77
48 张萍，申家恒，1998．高山红景天胚胎学研究．植物学研究，18(1)：39～45
49 张淑娟，刘与明，1996．洋桔梗F_1无菌播种和试管苗的快速繁殖．亚热带植物通讯，25(2)：13～16
50 张彦萍，1998．切花新秀—洋桔梗．植物杂志，2：20
51 赵梁军，2002．观赏植物生物学．北京：中国农业大学出版社，58～65
52 赵素君，韩志慧，孟广云，等，2003．洋桔梗在天津地区的栽培试验．天津农学院学报，10(1)：18～20
53 周根余，沈光华，程磊，2000．洋桔梗叶片培养中的全息现象．植物生理学通讯，36(3)：212～216
54 卓仁英，姜景民，2001．花木育种和繁殖技术研究概述．浙江林业科技，21(6)：6～10
55 祖元刚，张文辉，阎秀峰，等，1998．裂叶沙参与泡沙参解剖生态学的比较研究．植物研究，18(1) 46～54
56 Bradley J M, Rains S R, Manson J L et al., 2000. Flower pattern stability in genetically modified lisianthus
(Eustoma grandiflorum) under commercial growing conditions. New Zealand Journal of Crop and Horticultural Science,28(3): 175-184
57 Chao C H, Chen C C, Chang C A et al., 2000. Identification of a turnip mosaic virus isolate causing systemic yellow spotting on lisianthus. Plant Pathology Bulletin, 9(3): 115-122
58 Chao, Y. C., W. J. Liang, L. L. Huang et al., 1995. Bacterial wilt of Texas blue bell (Eustoma grandiflorum L.). Plant Pathol, 4:193-195
59 Chen C C, Chen Y K, Hsu H T, 2000. Characterization of a virus infecting lisianthus. Plant Disease, 84(5): 506-509
60 Chen H Y, Yang CH, 2000. Molecular characterization of MADS box genes regulating ovule evelopment and floral organ formation from Eustoma grandiflorum. Annual Meeting of The American Society of Plant Physiologists, 15:39
61 Cheong ChangHo, Yu KeeWon, Paek KeeYoeup et al., 1999. Randomly amplified polymorphic DNA (RAPD) analysis of the lisianthus (Eustoma grandiflorum Shinn.) variants obtained during tissue culture. Korean Journal of Horticultural Science and Technology, 17(3): 352-354
62 Cohen J, Lapidot M, Loebenstein G et al., 2001. First report of Sweet potato sunken vein virus occurring in Lisianthus.Plant Disease, 856(6): 79.
63 Derles S, Bradley M, Daviesk et al., 1995. Generation of novel pattens in Lisianthus flower using an antisense chalcone synthase. Acta Hortic, 420:26-28
64 Deroles S C, Bradley J M, Schwinn K E et al., 1998. An antisense chalcone synthase cDNA leads to novel colour patterns in lisianthus (Eustoma grandiflorum) flowers. Molecular Breeding, 4(1): 59-66
65 Hirai H, Mori G, 1999. The utilization of a spot air conditioner in raising of seedlings for rosetting prevention in Eustoma and Delphinium. Environment Control in Biology, 37(3): 191-196
66 Hisamatsu T, Koshioka M, Nishijima T et al., 1998. Identification of endogenous gibberellins and their role in rosetted seedlings of Eustoma grandiflorum. Journal of the Japanese Society for Horticultural Science, 67(6): 866-871
67 Huh KunYang, Bang Chang Seok, Song Jeong Seob et al., 1997. Effect of low temperature raising on flowering and cut flower quality of Eustoma grandiflorum. RDA Journal of Horticulture Science., 39(1): 107-111
68 Huh KunYang, Bang Chang Seok, Song Jeong Seob et al., 1998. Effect of long day treatments on the flowering of Eustoma grandiflorum raised at low temperature. RDA Journal of Horticulture Science Ⅱ, 40(2): 77-81
69 Ichimura K, 1998. Improvement of postharvest life in several cut flowers by the addition of sucrose. Japan Agricultural Research Quarterly, 32(4): 275-280
70 Ichimura K, Goto R, 2000. Acceleration of senescence by pollination of cut 'Asuka no nami' Eustoma flowers.
Journal of the Japanese Society for Horticultural Science, 69:166-170
71 Ichimura K, Korenaga M, 1998. Improvement of vase life and petal color expression in several cultivars of cut Eustoma flowers using sucrose with 8 hydroxyquinoline sulfate. Ornamental Plants and Tea, 13:31-39
72 Ichimura K, Shimamura M., Hisamatsu T, 1998. Role of ethylene in senescence of cut Eustoma flowers. Postharvest Biology and Technology, 14(2): 193-198
72 Kawabata S, Kusuhara Y, Li Y et al., 1999. The regulation of anthocyanin biosynthesis in Eustoma grandiflorum under low light conditions. Journal of the Japanese Society for Horticultural Science, 68(3): 519-526
73 Kritzman A, Beckelman H, Alexandrov S et al., 2000. Lisianthus leaf necrosis: a new disease of lisianthus caused by Iris yellow spot virus. Plant Disease, 84(11 ): 1185-1189
74 Lee JongWon, Kim TaeJoung, Kim JuHyoung et al., 2000. Effects of plant growth regulators on the growth and cut flower quality of Eustoma grandiflorum cv. Plantina Violet, 41(4): 419-422
75 Liao LiJen, Lin YuHan, Huang KuangLiang et al.,2001.Vase life of Eustoma grandiflorum as affected by aluminum sulfate. Botanical Bulletin of Academia Sinica, 42(1): 35-38
76 Markham K R, 2001. Cytoplasmic accumulation of flavonoids in flower petals and its relevance to yellow flower colouration. Phytochemistry, 58 (3), 403-416
77 Markham K R, Gould K S, Winefield C S et al., 2000. Anthocyanic vacuolar inclusions their nature and significance in flower colouration. Phytochemistry, 55(4): 327-336
78 Markham K R, Ryan K G, Gould K S et al., 2000. Cell wall sited flavonoids in lisianthus flower petals. Phytochemistry, 54(7): 681-687
79 McGovern R J, Bouzar H, Harbaugh B K, 2000a. Stem Blight of Eustoma grandiflorum caused by Sclerotium rolfsii. Plant Disease, 84(4): 490
80 McGovern R J, Seijo T E, Harbaugh B K et al., 2000b. Outbreaks of stem and leaf blight of Eustoma grandiflorum caused by a Phomopsis sp. in Florida. Plant Disease, 84(4): 491
81 McMillan R T Jr, Graves W, 2000. Impatiens necrotic spot tospo virus. Proceedings of the Florida State Horticultural Society, 112:290-291
82 Mercuri Antonio, Sacchetti Andrea, Benedetti Laura De, 2002. Green fluorescent flowers. Plant Science, 162 (4):647-655
83 Mino M, Oka Y, Yasaka M et al., 2003. Thermoinduction of genes encoding the enzymes of gibberellin biosynthesis and a putative negative regulator of gibberellin signal transduction in Eustoma grandiflorum. Plant Cell Reports, 22(2):159-163
84 Mototsugu Yanagida, Masanobu Mino, Masaki Iwabuchi et al., 2004. Reduced Glutathione is a Novel Regulator
of Vernalization Induced Bolting in the Rosette Plant Eustoma grandiflorum. Plant Cell Physiol, 45:129-137
85 Nielsen K M, Podivinsky E, 1997. cDNA cloning and endogenous expression of a flavonoid 3'5' hydroxylase from petals of lisianthus (Eustoma grandiflorum).Plant Science Limerick, 129(2): 167 -174
86 Ohkawa K, Maloupa E, 2000. Flower industry in Northeast Asia: development and introduction of new crops. Acta Horticulturae, 541 : 125-133
87 Ohkawa K, Sasaki E, Fischer G (ed.) et al., 1999. Eustoma (lisianthus) --its past, present, and future. Acta Horticulturae, 482:423-426
88 Ohki S, Sakai Y, Ooba K et al., 2000. Use of ion beams for in vitro mutation breeding of ornamental plants application in Eustoma grandiflorum. Acta Horticulturae, 508:75-82
89 Ohta K, Atarashi H, Shimatani Y et al., 2000. Effects of chitosan with or without nitrogen treatments on seedling growth in Eustoma grandiflorum (Raf.) Shinn. cv. Kairyou Wakamurasaki. Journal of the Japanese Society for Horticultural Science, 69(1): 63-65
90 Ohta K, Taniguchi A, Konishi N et al., 1999. Chitosan treatment affects plant growth and flower quality in Eustoma grandiflorum. HortScience, 34(2): 233-234
91 Oka M, Tasaka Y, Iwabuchi M et al., 2001. Elevated sensitivity to gibberellin by vernalization in the vegetative rosette plants of Eustoma grandiflorum and Arabidopsis thaliana. Plant Science., 160:(6):1237 - 1245
92 Oren Shamir M., Shaked Sachray L, Nissim Levi A et al., 1999. Anthocyanin pigmentation of lisianthus flower petals. Plant Science Limerick, 140(1 ): 81-86
93 Pecchia S, Forti M, Fanti S, et al., 2000. Outbreaks of Fusarium avenaceum on lisianthus (Eustoma russelianum) in Tuscany.Informatore Fitopatologico, 50(3): 61-64
94 Pergola G, Incrocci L, Serra F, 1998. Cold treatment of Eustoma russellianum seeds and seedlings: effects on growth and flowering. Italus Hortus, 5(3): 25-29
95 Rivas E B, Galleti S R, Duarte L M L et al., 2000. Virus and phytoplasm diseases of lisianthus.Summa Phytopathologica, 26(2): 257-262
96 Semeria L, Ruffoni B, Rabaglio M, 1996. Genetic transformation of Eustoma grandiflorum by Agrobacterium tumefaciens. Plant Cell, 67(1) :6
97 Shemluk M P, Kevan P G, 2001. Black flower colouration in wild Lisianthus nigrescens - its chemistry and ecological consequences. Plant Physiology and Biochemistry, 140:31-39
98 STEFAN VINCKIER, ERIK SMETS, 2003. Morphological and Ultrastructural Diversity of Orbicules in Gentianaceae, Ann. Bot., 92: 657-672.
99 Takahashi M, Nishihara M, Yamamura S, et al., 1998. Stable transformation of Eustoma grandiflorum by particle bombardment.Plant Cell Reports, 17(67): 504-507
100 Takezaki A, Fujino M, Nonaka M et al., 2000. The effects of temperature treatments on stem length of Eustoma grandiflorum. Acta Horticulturae, 515:151-157
101 Tanigawa T, Kobayashi Y, Matsui H et al., 2001. Effects of seedling age and high and low growth temperatures on bolting of Eustoma grandiflorum (Raf.) Shinn. cultivars. Journal of the Japanese Society for Horticultural Science, 70 (4): 501-509
102 Toppe B, Aanonsen E, Klemsdal S, 2004. First Report of Root Rot and Stem Blight Caused by Phytophthora palmivora in Eustoma grandiflorum. Plant Disease, 88(2): 224-228
103 Tsai Yu Tzeng, Hsing Yu Chen, Chang Hsien Yang, 2002. Ectopic Expression of Carpel Specific MADS Box Genes from Lily and Lisianthus Causes Similar Homeotic Conversion of Sepal and Petal in Arabidopsis. Plant Physiology, 130:1827-1836
104 Tsay JyhShyan, Kuo YinKang, Jeng YuShiow et al.,1999. Ultrastructural changes in the petals of senescing flowers of Eustoma russellianum. Journal of the Chinese Society for Horticultural Science, 45(4): 305-316
105 Uddin A F M, Jamal Hashimoto, Fumio Miwa, 2004. Seasonal variation in pigmentation and anthocyanidin phenetics in commercial Eustoma flowers. Scientia Horticulturae, 100(14), 103-116
106 Uddin AFMJ, Hashimoto F, Kaketani M et al., 2001. Analysis of light and sucrose potencies on petal coloration and pigmentation of lisianthus cultivars (in vitro). Scientia Horticulturae, 89(1): 73-82
107 Wolcan S, Lori G, Ronco L, 2001. First report of Fusarium solani causing stunt on lisianthus. Plant Disease, 85(4): 443
108 Yang H C, Hsieh T F, 1998. The occurrence and chemical control of downy mildew on Eustoma grandiflorum Salisb. Plant Protection Bulletin Taipei, 40(1): 37-48
109 Zaccai Michele, Edri Nurit, 2002. Floral transition in lisianthus (Eustoma grandiflorum). Scientia Horticulturae, 95(2): 333-341
2 [日]大川清，2000．花专科育种栽培Eustoma．诚文堂新光社，10～14
3 包满珠，1991．植物花青素基因的克隆及应用．园艺学报，24(3)：279～284
4 程金水，2001．园林植物遗传育种学．北京：中国林业出版社
5 方开泰，全辉，陈庆云，1988．实用回归分析．北京：科学出版社，114～116
6 傅荣昭，马江生，曹光诚，等，1995．观赏植物色香形基因工程研究进展．园艺学报，22(4)：381～385
7 高信曾，1986．植物学试验指导(形态解剖部分)．北京：高等教育出版社，58～179
8 郝立勤，顾金凤，2002．云南花卉产业发展对策研究．云南科技管理，4：12～15
9 何凤仙，2000．植物学试验．北京：高等教育出版社，41
10 何廷农，陈世龙，2000．条纹龙胆的胚胎学研究．西北植物学报，20(6)：960～967
11 何廷农，刘建全，1999．线叶龙胆的胚胎学研究．西北植物学报，19(2)：234～240
12 何奕昆，1995．雁来红叶色转变与超微结构及色素含量的关系．四川师范学院学报，16(3)：86～90
13 黄素梅，蓝福生，姚军，等，2000．我国花卉产业现状及其发展前景探讨．广西植物，20(3)：277～283
14 黄胤怡，沈明山，李鹏，等，2002．蓝色花基因工程．植物生理学通讯，38(2)：203～206
15 及华，温春秀，高延厅，1996．洋桔梗的离体快速繁殖，32(6)：431～432
16 及华，1999．日本的花卉业及生产经营．河北科技．2：38～41
17 金波，东惠茹，1994．一品红花色的探讨．园艺学报，21(1)：87～90
18 李嘉钰．1999，中国牡丹与芍药．北京：中国林业出版社，103～116
19 李懋学，张敩方，1991．植物染色体研究技术．哈尔滨：东北林业大学出版社，1～58
20 李正理，张新英，1987．植物解剖学．北京：高等教育出版社，166～293
21 林功涛，李凤宜，2000．北京小菊品系选育及遗传规律探讨．中国花卉科技二十年，北京：科学技术出版社，434～444
22 刘穆，2000．种子植物形态解剖学导论．北京：科学出版社，98～319
23 刘青林，陈俊愉，1998．观赏植物花器官主要观赏性状的遗传与改良．园艺学报，25(1)：81～86
24 潘会堂，张启翔，2000．花卉种质资源与遗传育种研究进展．北京林业大学学报，22(1)：81～86
25 潘家驹，1992．作物育种学总论．北京：中国农业大学出版社，50～56
26 庞志中，2000．花色研究概况．北京农业科学，18(5)：34～387
27 宋晓荭，文绍金，韩文婷，等，2003．借鉴经验，强化特色，加快结构调整步伐．青海农技推广，2：61～62
28 孙振雷，1999．观赏植物育种学．北京：民族出版社，45～49
29 孙自然，游捷，2000．我国菊花新品系选育进展．中国花卉科技二十年，北京：科学技术出版社，414～423
30 唐道城，张志英，赵梁军，2001．一串红花芽形态分化进程．园艺学报，28(4)：367～369
31 陶嘉龄，郑光华，1991．种子活力．北京：科学出版社，108～110
32 王瑗玲，1998a．洋桔梗的栽培(一)．花木盆景，25(1)：26
33 王瑷玲，1998b．洋桔梗的栽培(二)．花木盆景，26(2)：22～23
34 王钦丽，卢龙斗，吴小琴，2002．花粉的保存及其生活力测定．植物学通报，19(3)：365～373
35 王少平，朱爱民，2002．浸种对波斯菊种子发芽的影响．种子，108(2)：39～40
36 王仲礼，刘林德，田伟国，1998．短柄五加大、小孢子发生和雌雄配子体发育的研究．植物研究，18(2)：177～183
37 王仲礼，刘林德，田伟国，1999．短柄五加胚和胚乳发育的研究．植物研究，19(1)：41～46
38 吴昌陆，胡南珍，1995．蜡梅花部形态和开花习性研究．园艺学报，22(3)：277～282
39 吴人坚，1987．植物学试验方法．上海：上海科学技术出版社，52～77
40 夏宜平，2001．鲜切花培育技术．上海：上海科学技术出版社，85～86
41 徐克学，1999．生物数学．北京：科学出版社，40～49
42 徐振华，郭伟珍，王鑫，1997．洋桔梗的种子育苗技术．河北林业科技，4：43
43 薛淮，刘敏，张纯花，等，2002，花卉分子育种研究进展．生物工程进展，22(2)：80～83
44 杨继，2000．植物生物学试验．北京：高等教育出版社，施普林格出版社，67～117
45 杨霞，徐康，1997．洋桔梗的组织培养及快速繁殖．植物生理学通讯，33(6)：435～438
46 杨永刚，周根余，2001．洋桔梗叶片体外再生系统的激素调控．上海师范大学学报(自然科学版)，3(1)：99～100
47 余地求，李宝健，1997．花色素苷生物合成的遗传和发育调控．植物生理学通讯，33(1)：71～77
48 张萍，申家恒，1998．高山红景天胚胎学研究．植物学研究，18(1)：39～45
49 张淑娟，刘与明，1996．洋桔梗F_1无菌播种和试管苗的快速繁殖．亚热带植物通讯，25(2)：13～16
50 张彦萍，1998．切花新秀—洋桔梗．植物杂志，2：20
51 赵梁军，2002．观赏植物生物学．北京：中国农业大学出版社，58～65
52 赵素君，韩志慧，孟广云，等，2003．洋桔梗在天津地区的栽培试验．天津农学院学报，10(1)：18～20
53 周根余，沈光华，程磊，2000．洋桔梗叶片培养中的全息现象．植物生理学通讯，36(3)：212～216
54 卓仁英，姜景民，2001．花木育种和繁殖技术研究概述．浙江林业科技，21(6)：6～10
55 祖元刚，张文辉，阎秀峰，等，1998．裂叶沙参与泡沙参解剖生态学的比较研究．植物研究，18(1) 46～54
56 Bradley J M, Rains S R, Manson J L et al., 2000. Flower pattern stability in genetically modified lisianthus
(Eustoma grandiflorum) under commercial growing conditions. New Zealand Journal of Crop and Horticultural Science,28(3): 175-184
57 Chao C H, Chen C C, Chang C A et al., 2000. Identification of a turnip mosaic virus isolate causing systemic yellow spotting on lisianthus. Plant Pathology Bulletin, 9(3): 115-122
58 Chao, Y. C., W. J. Liang, L. L. Huang et al., 1995. Bacterial wilt of Texas blue bell (Eustoma grandiflorum L.). Plant Pathol, 4:193-195
59 Chen C C, Chen Y K, Hsu H T, 2000. Characterization of a virus infecting lisianthus. Plant Disease, 84(5): 506-509
60 Chen H Y, Yang CH, 2000. Molecular characterization of MADS box genes regulating ovule evelopment and floral organ formation from Eustoma grandiflorum. Annual Meeting of The American Society of Plant Physiologists, 15:39
61 Cheong ChangHo, Yu KeeWon, Paek KeeYoeup et al., 1999. Randomly amplified polymorphic DNA (RAPD) analysis of the lisianthus (Eustoma grandiflorum Shinn.) variants obtained during tissue culture. Korean Journal of Horticultural Science and Technology, 17(3): 352-354
62 Cohen J, Lapidot M, Loebenstein G et al., 2001. First report of Sweet potato sunken vein virus occurring in Lisianthus.Plant Disease, 856(6): 79.
63 Derles S, Bradley M, Daviesk et al., 1995. Generation of novel pattens in Lisianthus flower using an antisense chalcone synthase. Acta Hortic, 420:26-28
64 Deroles S C, Bradley J M, Schwinn K E et al., 1998. An antisense chalcone synthase cDNA leads to novel colour patterns in lisianthus (Eustoma grandiflorum) flowers. Molecular Breeding, 4(1): 59-66
65 Hirai H, Mori G, 1999. The utilization of a spot air conditioner in raising of seedlings for rosetting prevention in Eustoma and Delphinium. Environment Control in Biology, 37(3): 191-196
66 Hisamatsu T, Koshioka M, Nishijima T et al., 1998. Identification of endogenous gibberellins and their role in rosetted seedlings of Eustoma grandiflorum. Journal of the Japanese Society for Horticultural Science, 67(6): 866-871
67 Huh KunYang, Bang Chang Seok, Song Jeong Seob et al., 1997. Effect of low temperature raising on flowering and cut flower quality of Eustoma grandiflorum. RDA Journal of Horticulture Science., 39(1): 107-111
68 Huh KunYang, Bang Chang Seok, Song Jeong Seob et al., 1998. Effect of long day treatments on the flowering of Eustoma grandiflorum raised at low temperature. RDA Journal of Horticulture Science Ⅱ, 40(2): 77-81
69 Ichimura K, 1998. Improvement of postharvest life in several cut flowers by the addition of sucrose. Japan Agricultural Research Quarterly, 32(4): 275-280
70 Ichimura K, Goto R, 2000. Acceleration of senescence by pollination of cut 'Asuka no nami' Eustoma flowers.
Journal of the Japanese Society for Horticultural Science, 69:166-170
71 Ichimura K, Korenaga M, 1998. Improvement of vase life and petal color expression in several cultivars of cut Eustoma flowers using sucrose with 8 hydroxyquinoline sulfate. Ornamental Plants and Tea, 13:31-39
72 Ichimura K, Shimamura M., Hisamatsu T, 1998. Role of ethylene in senescence of cut Eustoma flowers. Postharvest Biology and Technology, 14(2): 193-198
72 Kawabata S, Kusuhara Y, Li Y et al., 1999. The regulation of anthocyanin biosynthesis in Eustoma grandiflorum under low light conditions. Journal of the Japanese Society for Horticultural Science, 68(3): 519-526
73 Kritzman A, Beckelman H, Alexandrov S et al., 2000. Lisianthus leaf necrosis: a new disease of lisianthus caused by Iris yellow spot virus. Plant Disease, 84(11 ): 1185-1189
74 Lee JongWon, Kim TaeJoung, Kim JuHyoung et al., 2000. Effects of plant growth regulators on the growth and cut flower quality of Eustoma grandiflorum cv. Plantina Violet, 41(4): 419-422
75 Liao LiJen, Lin YuHan, Huang KuangLiang et al.,2001.Vase life of Eustoma grandiflorum as affected by aluminum sulfate. Botanical Bulletin of Academia Sinica, 42(1): 35-38
76 Markham K R, 2001. Cytoplasmic accumulation of flavonoids in flower petals and its relevance to yellow flower colouration. Phytochemistry, 58 (3), 403-416
77 Markham K R, Gould K S, Winefield C S et al., 2000. Anthocyanic vacuolar inclusions their nature and significance in flower colouration. Phytochemistry, 55(4): 327-336
78 Markham K R, Ryan K G, Gould K S et al., 2000. Cell wall sited flavonoids in lisianthus flower petals. Phytochemistry, 54(7): 681-687
79 McGovern R J, Bouzar H, Harbaugh B K, 2000a. Stem Blight of Eustoma grandiflorum caused by Sclerotium rolfsii. Plant Disease, 84(4): 490
80 McGovern R J, Seijo T E, Harbaugh B K et al., 2000b. Outbreaks of stem and leaf blight of Eustoma grandiflorum caused by a Phomopsis sp. in Florida. Plant Disease, 84(4): 491
81 McMillan R T Jr, Graves W, 2000. Impatiens necrotic spot tospo virus. Proceedings of the Florida State Horticultural Society, 112:290-291
82 Mercuri Antonio, Sacchetti Andrea, Benedetti Laura De, 2002. Green fluorescent flowers. Plant Science, 162 (4):647-655
83 Mino M, Oka Y, Yasaka M et al., 2003. Thermoinduction of genes encoding the enzymes of gibberellin biosynthesis and a putative negative regulator of gibberellin signal transduction in Eustoma grandiflorum. Plant Cell Reports, 22(2):159-163
84 Mototsugu Yanagida, Masanobu Mino, Masaki Iwabuchi et al., 2004. Reduced Glutathione is a Novel Regulator
of Vernalization Induced Bolting in the Rosette Plant Eustoma grandiflorum. Plant Cell Physiol, 45:129-137
85 Nielsen K M, Podivinsky E, 1997. cDNA cloning and endogenous expression of a flavonoid 3'5' hydroxylase from petals of lisianthus (Eustoma grandiflorum).Plant Science Limerick, 129(2): 167 -174
86 Ohkawa K, Maloupa E, 2000. Flower industry in Northeast Asia: development and introduction of new crops. Acta Horticulturae, 541 : 125-133
87 Ohkawa K, Sasaki E, Fischer G (ed.) et al., 1999. Eustoma (lisianthus) --its past, present, and future. Acta Horticulturae, 482:423-426
88 Ohki S, Sakai Y, Ooba K et al., 2000. Use of ion beams for in vitro mutation breeding of ornamental plants application in Eustoma grandiflorum. Acta Horticulturae, 508:75-82
89 Ohta K, Atarashi H, Shimatani Y et al., 2000. Effects of chitosan with or without nitrogen treatments on seedling growth in Eustoma grandiflorum (Raf.) Shinn. cv. Kairyou Wakamurasaki. Journal of the Japanese Society for Horticultural Science, 69(1): 63-65
90 Ohta K, Taniguchi A, Konishi N et al., 1999. Chitosan treatment affects plant growth and flower quality in Eustoma grandiflorum. HortScience, 34(2): 233-234
91 Oka M, Tasaka Y, Iwabuchi M et al., 2001. Elevated sensitivity to gibberellin by vernalization in the vegetative rosette plants of Eustoma grandiflorum and Arabidopsis thaliana. Plant Science., 160:(6):1237 - 1245
92 Oren Shamir M., Shaked Sachray L, Nissim Levi A et al., 1999. Anthocyanin pigmentation of lisianthus flower petals. Plant Science Limerick, 140(1 ): 81-86
93 Pecchia S, Forti M, Fanti S, et al., 2000. Outbreaks of Fusarium avenaceum on lisianthus (Eustoma russelianum) in Tuscany.Informatore Fitopatologico, 50(3): 61-64
94 Pergola G, Incrocci L, Serra F, 1998. Cold treatment of Eustoma russellianum seeds and seedlings: effects on growth and flowering. Italus Hortus, 5(3): 25-29
95 Rivas E B, Galleti S R, Duarte L M L et al., 2000. Virus and phytoplasm diseases of lisianthus.Summa Phytopathologica, 26(2): 257-262
96 Semeria L, Ruffoni B, Rabaglio M, 1996. Genetic transformation of Eustoma grandiflorum by Agrobacterium tumefaciens. Plant Cell, 67(1) :6
97 Shemluk M P, Kevan P G, 2001. Black flower colouration in wild Lisianthus nigrescens - its chemistry and ecological consequences. Plant Physiology and Biochemistry, 140:31-39
98 STEFAN VINCKIER, ERIK SMETS, 2003. Morphological and Ultrastructural Diversity of Orbicules in Gentianaceae, Ann. Bot., 92: 657-672.
99 Takahashi M, Nishihara M, Yamamura S, et al., 1998. Stable transformation of Eustoma grandiflorum by particle bombardment.Plant Cell Reports, 17(67): 504-507
100 Takezaki A, Fujino M, Nonaka M et al., 2000. The effects of temperature treatments on stem length of Eustoma grandiflorum. Acta Horticulturae, 515:151-157
101 Tanigawa T, Kobayashi Y, Matsui H et al., 2001. Effects of seedling age and high and low growth temperatures on bolting of Eustoma grandiflorum (Raf.) Shinn. cultivars. Journal of the Japanese Society for Horticultural Science, 70 (4): 501-509
102 Toppe B, Aanonsen E, Klemsdal S, 2004. First Report of Root Rot and Stem Blight Caused by Phytophthora palmivora in Eustoma grandiflorum. Plant Disease, 88(2): 224-228
103 Tsai Yu Tzeng, Hsing Yu Chen, Chang Hsien Yang, 2002. Ectopic Expression of Carpel Specific MADS Box Genes from Lily and Lisianthus Causes Similar Homeotic Conversion of Sepal and Petal in Arabidopsis. Plant Physiology, 130:1827-1836
104 Tsay JyhShyan, Kuo YinKang, Jeng YuShiow et al.,1999. Ultrastructural changes in the petals of senescing flowers of Eustoma russellianum. Journal of the Chinese Society for Horticultural Science, 45(4): 305-316
105 Uddin A F M, Jamal Hashimoto, Fumio Miwa, 2004. Seasonal variation in pigmentation and anthocyanidin phenetics in commercial Eustoma flowers. Scientia Horticulturae, 100(14), 103-116
106 Uddin AFMJ, Hashimoto F, Kaketani M et al., 2001. Analysis of light and sucrose potencies on petal coloration and pigmentation of lisianthus cultivars (in vitro). Scientia Horticulturae, 89(1): 73-82
107 Wolcan S, Lori G, Ronco L, 2001. First report of Fusarium solani causing stunt on lisianthus. Plant Disease, 85(4): 443
108 Yang H C, Hsieh T F, 1998. The occurrence and chemical control of downy mildew on Eustoma grandiflorum Salisb. Plant Protection Bulletin Taipei, 40(1): 37-48
109 Zaccai Michele, Edri Nurit, 2002. Floral transition in lisianthus (Eustoma grandiflorum). Scientia Horticulturae, 95(2): 333-341