农杆菌介导NPR1基因对百合遗传转化的研究
摘要
百合(Lilium spp.)不但花姿独特,同时还具备药用和食用价值,深受国内外市场的喜爱。随着百合栽培面积的不断扩大,这种易感病的植物常常引起大面积的病害,造成巨大的经济损失。然而传统的药剂防治不但增加人工成本,还会对环境造成污染,应用抗病品种是病害防治的最直接有效的方法。但采用传统的育种方法不但周期较长,受亲本影响较大,因此这种育种方法不能满足生产的需要。随着植物生物技术的不断发展,直接将外源目的基因导入植物材料中,这为百合培育新品种开辟了新的道路。
本研究以抗病能力差的东方百合“Sorbonne”为试材,对东方百合鳞片进行了直接再生不定芽系统的建立,同时以鳞片为外植体进行了愈伤组织的诱导,进行了愈伤组织经体细胞胚发生途径再生的研究。并对直接再生过程中的生根情况进行了比较,对鳞片愈伤的诱导体胚发育进行了较为深入的研究。在此基础上以百合鳞片为遗传转化受体,以鳞片直接再生芽方式为再生途径,将植物广谱抗病基因(NPR1基因)通过农杆菌介导转化百合,建立了稳定的遗传转化体系,得到了经PCR检测为阳性的植株。本研究的主要结果如下:
1.百合鳞片直接再生不定芽体系:以百合鳞片为外植体,最佳芽诱导培养基为MS+6-BA0.4mg/L+NAA1.0mg/L,芽诱导率达到96.7%;最佳生根培养基为MS+IBA0.5mg/L,生根率为95.2%。
2.百合愈伤组织体细胞胚发生途径的再生体系:以百合鳞片作为外植体,最佳愈伤诱导培养基为MS+2,4-D 2.0mg/L +6-BA 0.5mg/L+水解酪蛋白300mg/L,诱导率为68.3%;最佳胚性愈伤诱导培养基为MS+2,4-D2.0mg/L +6-BA0.5mg/L+水解酪蛋白150mg/L+谷氨酰胺150mg/L,诱导率为75.0%;将其放入MS+2,4-D 0.5 mg/L +KT 0.3 mg/L+蔗糖40g/L中培养,体胚形成率为60.0%,最佳体细胞胚萌发培养基为MS+6-BA0.5 mg/L。通过石蜡切片法对鳞片愈伤的发育过程观察发现,首先外植体表层细胞形成愈伤组织,随后在愈伤组织表面形成许多瘤状凸起,这些凸起继续发育成球形胚、心形胚,最后发育成熟形成完整植株。
3.百合遗传转化体系:以百合鳞片为转化受体,在用农杆菌侵染前预培养2天,侵染时菌液浓度为OD600=0.5,侵染时间为20min,20℃下共培养时间2天,共培养之后进行Km浓度为30―80―130mg/L的依次筛选,头孢霉素在除菌过程中使用浓度为400mg/L,直至抗性芽的形成。通过实验发现在共培养时,去除培养基中NH4NO3,并向其中加入100mg/L的乙酰丁香酮可提高转化率。
4.抗性植株的PCR检测:由直接芽再生途径得到的56株抗性植株中有3株呈阳性,阳性率为5.4%,占该再生系统总转化外植体900块的0.33%。
Lilies(Lilium spp.)not only have special beautiful flowers, but also have officinal and edible value. They are very popular for many people in the world. With the planting areas expanded gradually, the flowers often bring lots of disease on the field which are sensitive for illness. So they produced hugeness lost. But tradition preventions increase labour cost,meanwhile they could effect the environment. It is more effective and direct method for preventing the plant diseases to use using the disease-resistant varieties. Because the tradition breeding method need a long time for breeding and is effected by F1 generation greatly, the demand of the Lilies production couldn’t be meted. On the background of biology technique fast development, we could transfer new disease-resistant gene of other sources into the Lilies by using genetic engineering means .It will break a new path in the new disease-resistant breeding of Lilies.
This study took the less-disease resistant oriental lily‘Sorbonne’as material. We establish regeneration system of direct shoot method, introduce callus from Lilies bulbs。We observe the transformation from callus to somatic embryo. We compare the cases of shoot on different medias in the process of direct regeneration. We use the bulbs of Lilies as the explants of genetic transformation, The broad-spectrum disease resistance gene (NPR1 gene) transferred into Lilies mediated by the agrobacterium. we establish genetic transformation system of Lilies. Finally, we get positive plants after PCR. The main content and result of experiment are as follows:
1.The regeneration system though bulbs: we use the slice of Lilies bulbs as explants, MS basal medium supplemented with 6-BA0.4mg/L and NAA1.0mg/L was advantageous to shoot induction, the frequency of shoot induction is 96.7%; MS basal medium supplemented with IBA0.5 mg/L could develop their roots, the rate of robust plantlets is 95.2%.
2.The callus regeneration of Lilies by somatic embryogenesis: we use the bulbs of Lilies as explants, the good method for induce callus is MS basal medium supplemented with 2,4-D2.0mg/L and 6-BA0.5 mg/L and LH 300mg/L, the rate of produce callus is 68.3%. MS+2,4-D 2.0 mg/L +6-BA 0.5mg/L+LH150mg/L+Gln150mg/L is the best culture medium that the induction of embryogenesis callus ,and the rate of produce embryogenesis callus is 75.0% .Then transfer the embryogenesis callus into the culture media MS+2,4-D 0.5 mg/L +KT 0.3 mg/L + Sucrose 40g/L, which rate is 60.0%. Somatic embryo could germinate on the MS basal medium supplemented with 6-BA 0.5mg/L. The appearance and growth of somatic embryo were observed through paraffin slices. Histological observations showed that surface cells first experienced dedifferentiation and followed with callus-forming, then the top of callus appear a tubercular structure. These embryogenic cell masses experienced globular, heart stages and developed into whole plantlets .
3.The establishment of genetic transformation system: Taking the slice of Lilies bulbs as explants, pre-culture of callus for 2d;the agrobacterium tumefaciens solution was OD600=0.5,the time of infect was 20min,the time of co-culture was 2d at 20℃,the step up the concentration of Km 30mg/L from the first day,80mg/L from the third day,130mg/L from the sixth day. Concentration of Cef was 400mg/L in the process of sterilization, until the plantlet formed. MS added 100mg/L trans-Rerulic acid and subtracted NH4NO3 can improve the transformation ratio.
4.Detection of the transformed plant:For shoot regeneration,we gained 56 resistant plants.Three of them is PCR positive plant, positive percentage is 5.4% and transformation percentage is 0.33% in 900 explants.
本研究以抗病能力差的东方百合“Sorbonne”为试材,对东方百合鳞片进行了直接再生不定芽系统的建立,同时以鳞片为外植体进行了愈伤组织的诱导,进行了愈伤组织经体细胞胚发生途径再生的研究。并对直接再生过程中的生根情况进行了比较,对鳞片愈伤的诱导体胚发育进行了较为深入的研究。在此基础上以百合鳞片为遗传转化受体,以鳞片直接再生芽方式为再生途径,将植物广谱抗病基因(NPR1基因)通过农杆菌介导转化百合,建立了稳定的遗传转化体系,得到了经PCR检测为阳性的植株。本研究的主要结果如下:
1.百合鳞片直接再生不定芽体系:以百合鳞片为外植体,最佳芽诱导培养基为MS+6-BA0.4mg/L+NAA1.0mg/L,芽诱导率达到96.7%;最佳生根培养基为MS+IBA0.5mg/L,生根率为95.2%。
2.百合愈伤组织体细胞胚发生途径的再生体系:以百合鳞片作为外植体,最佳愈伤诱导培养基为MS+2,4-D 2.0mg/L +6-BA 0.5mg/L+水解酪蛋白300mg/L,诱导率为68.3%;最佳胚性愈伤诱导培养基为MS+2,4-D2.0mg/L +6-BA0.5mg/L+水解酪蛋白150mg/L+谷氨酰胺150mg/L,诱导率为75.0%;将其放入MS+2,4-D 0.5 mg/L +KT 0.3 mg/L+蔗糖40g/L中培养,体胚形成率为60.0%,最佳体细胞胚萌发培养基为MS+6-BA0.5 mg/L。通过石蜡切片法对鳞片愈伤的发育过程观察发现,首先外植体表层细胞形成愈伤组织,随后在愈伤组织表面形成许多瘤状凸起,这些凸起继续发育成球形胚、心形胚,最后发育成熟形成完整植株。
3.百合遗传转化体系:以百合鳞片为转化受体,在用农杆菌侵染前预培养2天,侵染时菌液浓度为OD600=0.5,侵染时间为20min,20℃下共培养时间2天,共培养之后进行Km浓度为30―80―130mg/L的依次筛选,头孢霉素在除菌过程中使用浓度为400mg/L,直至抗性芽的形成。通过实验发现在共培养时,去除培养基中NH4NO3,并向其中加入100mg/L的乙酰丁香酮可提高转化率。
4.抗性植株的PCR检测:由直接芽再生途径得到的56株抗性植株中有3株呈阳性,阳性率为5.4%,占该再生系统总转化外植体900块的0.33%。
Lilies(Lilium spp.)not only have special beautiful flowers, but also have officinal and edible value. They are very popular for many people in the world. With the planting areas expanded gradually, the flowers often bring lots of disease on the field which are sensitive for illness. So they produced hugeness lost. But tradition preventions increase labour cost,meanwhile they could effect the environment. It is more effective and direct method for preventing the plant diseases to use using the disease-resistant varieties. Because the tradition breeding method need a long time for breeding and is effected by F1 generation greatly, the demand of the Lilies production couldn’t be meted. On the background of biology technique fast development, we could transfer new disease-resistant gene of other sources into the Lilies by using genetic engineering means .It will break a new path in the new disease-resistant breeding of Lilies.
This study took the less-disease resistant oriental lily‘Sorbonne’as material. We establish regeneration system of direct shoot method, introduce callus from Lilies bulbs。We observe the transformation from callus to somatic embryo. We compare the cases of shoot on different medias in the process of direct regeneration. We use the bulbs of Lilies as the explants of genetic transformation, The broad-spectrum disease resistance gene (NPR1 gene) transferred into Lilies mediated by the agrobacterium. we establish genetic transformation system of Lilies. Finally, we get positive plants after PCR. The main content and result of experiment are as follows:
1.The regeneration system though bulbs: we use the slice of Lilies bulbs as explants, MS basal medium supplemented with 6-BA0.4mg/L and NAA1.0mg/L was advantageous to shoot induction, the frequency of shoot induction is 96.7%; MS basal medium supplemented with IBA0.5 mg/L could develop their roots, the rate of robust plantlets is 95.2%.
2.The callus regeneration of Lilies by somatic embryogenesis: we use the bulbs of Lilies as explants, the good method for induce callus is MS basal medium supplemented with 2,4-D2.0mg/L and 6-BA0.5 mg/L and LH 300mg/L, the rate of produce callus is 68.3%. MS+2,4-D 2.0 mg/L +6-BA 0.5mg/L+LH150mg/L+Gln150mg/L is the best culture medium that the induction of embryogenesis callus ,and the rate of produce embryogenesis callus is 75.0% .Then transfer the embryogenesis callus into the culture media MS+2,4-D 0.5 mg/L +KT 0.3 mg/L + Sucrose 40g/L, which rate is 60.0%. Somatic embryo could germinate on the MS basal medium supplemented with 6-BA 0.5mg/L. The appearance and growth of somatic embryo were observed through paraffin slices. Histological observations showed that surface cells first experienced dedifferentiation and followed with callus-forming, then the top of callus appear a tubercular structure. These embryogenic cell masses experienced globular, heart stages and developed into whole plantlets .
3.The establishment of genetic transformation system: Taking the slice of Lilies bulbs as explants, pre-culture of callus for 2d;the agrobacterium tumefaciens solution was OD600=0.5,the time of infect was 20min,the time of co-culture was 2d at 20℃,the step up the concentration of Km 30mg/L from the first day,80mg/L from the third day,130mg/L from the sixth day. Concentration of Cef was 400mg/L in the process of sterilization, until the plantlet formed. MS added 100mg/L trans-Rerulic acid and subtracted NH4NO3 can improve the transformation ratio.
4.Detection of the transformed plant:For shoot regeneration,we gained 56 resistant plants.Three of them is PCR positive plant, positive percentage is 5.4% and transformation percentage is 0.33% in 900 explants.
引文
鲍惟钊,吴敬音,王月芳等. 1984.百合组培诱变研究器官水平的多倍体诱变.江苏农业科学. (1):28～30
曾镭,刘燕.2007.植物组织培养中褐化问题的研究进展.安徽农学通报.13(14):49～50
陈秋萍.2002.福建省百合病害调查初报.福建林学院学报.20(2):162～164
陈学林,李卫锋,黄海涛.2007.荷兰百合索尔邦的离体组织培养.安徽农业科学.35(20):6045～6046
陈以峰.1996.植物体细胞胚胎发生的诱导与调节研究进展.生物学杂志.69(1):3～6
程爱芳.2006.百合(Lilium spp)的组培快速繁殖技术研究.武汉生物工程学院学报.2(1):27-29
程金水.2001.园林植物遗传育种学.中国林业出版社.309～312
程佑发,王勋陵.2001.枣树体细胞胚发生的组织学研究.西北植物学报.211: 142～145
褚云霞,张永春,杨红娟等.2002.百合花药愈伤组织诱导中的基因型差异.上海农业学报.18(3):17～20
邓晓玲,孔维文,冯震,郭明,李华平. 2004.甘蓝型油菜中NPR1基因的克隆、载体构建及其转化的研究.2004年中国植物病理学会年会论文集:570～571
丁兰,刘国安,田卫东. 2005.新铁炮百合组织培养和快速繁殖研究.西北师范大学学报.37(1): 80～82
范慈惠. 1981.不同光源对百合珠芽培养的影响.江苏农业科学.3:205～207
方华舟,洪万泓.2006.百合组织培养及快繁技术的研究与探讨.贵州林业科学.34(4): 27～29
郝建平,金竹萍,王永康等.2006.枣合子胚和体细胞胚发育过程的观察与比较.分子细胞生物学报.39(5):423～426
贺晓庆.2007.NPKl和NPR1基因的克隆及其转化辣椒的研究.西北农林科技大学硕士论文
胡凤仙. 2003.转拟南芥系统获得抗性调节基因(NPRl)烟草的获得及其遗传分析.广西大学硕十学位论文
胡风荣,席梦利,刘光欣,池坚.东方百合的器官发生与体胚发生的研究.南京林业大学学报(自然科学版).31(2):5～8
黄济明,赵晓艺,张国民等. 1990.玫红百合为亲本育成百合种间杂种.园艺学报.17 (2):153～156
黄济明. 1982.王百合×大卫百合种间远源杂种的育成.园艺学报.9(3):51～56
郎楷永. 2003.中国的百合.科技导报.7:24,37～40
李铁军,郁伟. 2002.百合主要病害的识别与防治.农业科技通讯.(1):30～31
李小军. 2005.农杆菌介导NPR1基因转化安祖花的研究.上海师范大学硕士论文
梁巧兰,徐秉良.2004.观赏百合根腐病病原鉴定及药剂筛选.甘肃农业大学学报.39(1):25～28
刘芬,王发林. 2001.兰州百合花丝组培诱导完整植株的研究.甘肃农业科技.6: 29～31
刘何,谢令琴,赵建军.2007.植物抗性相关基因的研究.华北农学报.22:17～20
刘菊华,金志强,徐碧玉等.2003.龙牙百合的遗传转化及再生.分子植物育种. 1(4):465～474
刘明志,林雪艳. 2002.激素对百合植株再生的影响.广西植物.22(2): 167～170
刘选明,周朴华,屈妹存等.1997.百合鳞片叶离体诱导形成不定芽和体细胞胚.园艺学报.24(4):353～358
卢其能. 2002.龙牙百合的组织培养和快速繁殖研究.江西农业学报.14(4): 43～46
陆家云.1995.药用植物病害.中国农业出版社.45～68
罗荡平. 2004.白叶枯病和稻瘟病拟南芥NPR1基因转入水稻使水稻获得广谱抗病性.广西大学硕士学位论文
罗凤霞,徐桂华,金丽丽等. 2000.新铁炮百合微繁的研究.沈阳农业大学学报.31(3): 254～257
罗其德. 2004.Genetic Modification to Enhance Tomato Resistance to Disease.广西大学硕士论文
毛元荣. 2004.洋桔梗叶片再生体系的建立及农杆菌介导的NPR1基因转化洋桔梗的初步研究.上海师范大学硕士论文
莫昭展,符韵林,戚萌. 2007.野百合鳞茎芽的诱导和增殖的初步研究.安徽农业科学. 35(31):9890～9892
欧阳秩,吴帮承.1996.观赏植物病害.中国农业出版社.23～65
乔永旭,陈超,张永平等.2007.东方百合“索邦”离体培养再生体系的建立.安徽农业科学.35(1):67～69
沈淑琳.1996.百合病毒及其检验.植物检疫.10(4):223～225
衰澍,贾勇炯,林宏辉. 2003.诱导植物体细胞胚发生的几个生理因素.植物生理学通讯.39(5):508～512.
唐东芹,钱虹妹,唐克轩等.2004.根癌农杆菌介导半夏凝集素基因pBIXPTA对百合的转化.上海交通大学学报.22(2):135～137
唐东芹,钱虹妹等. 2003.百合基因转化胚性愈伤组织受体系统的建立.浙江林学院学报.20(3):273～276
唐祥宁,游春平,刘福秀等.1997.江西百合病害调查与鉴定.江西农业学报.9(4):l～8
唐益苗. 2006.小麦及其近缘植物NPR1类似基因和反转录转座子基因片段的分离和特性分析.中国农业科学院博士论文
汪海洋,张慧珍,李炳武等.2002.百合病害的发生与防治.上海蔬菜.4:35～36
王爱勤,周歧伟,何龙飞等.1998.百合试管结鳞茎的研究.广西农业大学学报.17(1):71～75
王刚,杜捷,李桂英等. 2002.兰州百合和野百合组织培养及快速繁殖研究.西北师范大学学报(自然科学版).38(1): 69～71
王关林,方宏筠等. 2002.抗菌肽基因转化大白菜获得抗病转基因植株及稳定遗传.植物学报.44(8):951～955
王季林. 1987.百合鳞片不同部位繁殖系数的观察.江苏农业科学.1: 30
王清连,王敏,师海荣. 2005.植物激素对棉花体细胞胚胎发生的诱导及调节作用.生物技术通讯.15(6):577～579
王仁祥.2005.转基因植物研究进展.生物学教学.30(13):9～12
温春秀,及华. 1997.新铁炮百合((L. longiflorum×L. formosanum)的离体快速繁殖.植物生理学通讯.33(3): 198
吴乃虎编著. 2002.基因工程原理(第二版下册).北京:科学出版社.171～225.
吴若普,阮少宁等. 2003.百合胚状体形成途径及植株的再生研究.福建林学院学报. 23(4):322～325
徐碧玉,刘菊华,金志强.2005.类黄酮3',5'羟基化酶基因的克隆及转化铁炮百合.园艺学报.
许宝辉. 2002.百合子房组织培养研究.南科技大学学报.3:35～38
杨乃博. 1981.十二种植物的试管无性繁殖.植物学报.23(4):284
姚祥坦,曹家树,李晋豫,王神云.2004.植物抗病毒病育种策略.细胞生物学杂志.26:362～366
叶世森,林芳,宋建英. 2005.百合病害的研究综述.西南林学院学报.25(3):84～88
于湄,叶长明. 1999.转基因植物中RNA介导病毒抗性.植物学通报.16(2) :127～130
张德水,陈受宜. 1997.植物抗病性的分子生物学研究进展.植物病理学报.27(2):97～103
张丽珍.2005.NPR1基因的克隆及西瓜的遗传转化的研究.广西师范大学硕士论文.
张兴翠,周昌华,殷家明等. 2003.药用百合的多倍体诱导及快速繁殖.西南农业大学学报.25(1):14～17
张艳贞,魏松红,杜娟,王罡. 2000.植物抗病分子机制及抗病基因工程研究进展.沈阳农业大学学报.31(4):365～369
张艺萍,屈云慧,王祥宁等.2007.提高百合茎尖组织培养脱毒效率研究.广东农业科学.1:37～38
张于光,肖启明.2003.百合的主要病害及其防治.植物杂志.(3):16～17
张云,原雅玲,刘青林. 2001.百合品种改良与生物技术研究进展.北京林业大学学报. 23(6): 56～59
张中义. 1992.观赏植物真菌病害.四川科技出版社.26～57
赵庆芳,曾小英,丁兰等. 2003.东方百合组织培养和快速繁殖研究.西北师范大学学报(自然科学版).39(1):66～68
张玉静,王志刚,印东生,王平.2007.百合主要病害及防治.北方园艺.11:202
张志兰.2007.山茶花组织培养过程忠的防外植体褐化试验.西部林业科学.36(2):118～121
郑秀芳,谬为. 2002.抗菌肤及在植物抗病基因工程中的应用.生物学通报.37(10): 9～10
钟景辉,蔡秋锦,张飞萍等.2000.百合病害及其持续治理.森林病虫通讯.19(2):28～31,23.
赵欢蕊,刘雅莉,王跃进.2007.农杆菌介导的普利安娜百合鳞茎直接分化受体系统的建立.西北农业学报.16(4):142～147
周俊彦.1981.植物体细胞在组织培养中产生的胚状体I植物体细胞的胚状体发生.植物生理学报.7(4):391～395
周晓燕. 2002.百合主要病害及其防治.植物保护.28(1):57～58
周艳萍,郑红娟,贾桂霞. 2007.两个亚洲百合品种离体再生体系的建立.北京林业大学学报.29(1):123～127
诸葛强,王婕琛,陈英等. 2003.新疆杨高效遗传转化系统的建立.植物资源与环境学报. 12(4):6～10
邹新慧,何平.2002.植物抗病毒基因工程研究进展及存在的问题.江西农业报. 14(4):59～65
A. A. Watad,D.-J. Yun,T. Matsumoto,X. Niu,Y. Wu,A. K. Kononowicz,R. A. Bressan,P. M. Hasegawa . 1998 . Microprojectile bombardment-mediated transformation of Lilium longiflorum.Plant Cell Reports.17: 262～267
Adam M,Dimech,Rob Cross,Rebecca Ford.2007.Micropropagation of Gymea Lily (Doryanthes excelsa Correa) from New South Wales, Australia.Plant Cell, Tissue and Organ Culture.88: 157～165
Akira Kikuchi, Nobuya Sanuki, Katsumi Higashi.2006.Abscisic acid and stress treatment are essential for the acquisition of embryogenic competence by carrot somatic.Planta.223:637~645
Arzate F A M,Nakazaki T,Okumoto Y et al.1997.Efficient callus induction and plant regenration from filaments with anther in lily (L.longiflorum).Plant Cell Reports.16:836～840 BELLARDI M G,NANNI G.2002.Old and new viruses of lily in Italy.Acta Horticulturae.568:215～220
Cao. H.,Li. X.,Dong .X.1998.Generation of broad-spectrum disease resistance by over expression of an essential regulatory gene in systemic acquired resistance.Proc Natl Acad Sci USA.95:6531～6536
Carr JP,Gal DNA et al.1994.Replicase-mediated resistance to cucumber mosaic virus in transgenic plants involves suppression of both virus replication in the inoculated leaves and long distance move meat.Virology.199:439～447
Chern. M., Canlas.P.E., Fitzgerald. H. A., Ronald. P. C. 2005.Rice NRR, a negative regulator of disease resistance,interacts with Arabidopsis NPR1 and rice NH1.Plant Journal.43: 623～35
Chern. M.,Fitzgerald. H.A.,Canlas. P. E.,Navarre. D. A.,Ronald. P. C.2005. Over expression of a rice NPR1 homolog leads to constitutive activation of defense response and hypersensitivity to light. Mol Plant Microbe Interact.18: 511～520
Chern. M.S.,Fitzgerald. H.A.,Yadav. R.C.,Canlas. P. E..et al.2001.Evidence for a disease-resistance pathway in rice similar to the NPR1-mediated signaling pathway in Arabidopsis.Plant Journal.27:101～113
COHEN A,MEREDITHC P,SANIEWSKIM et al .1992.A grobacterium-mediated transformation of Lilium[J].ActaH ort.325:611～618
COHEN J,GERA A.1996.Virus diseases of lilies in Israel[J].Acta Horticulture.432:84～87
Cooper B,Lapidot M et al.1995.A defective movement protein of TMV in transgenic plants confers resistant to multiple viruses whereas the functional analog in creases susceptibility[J]. Virology.206:307～313
Cossio F. et al.1985.Further findings on the in vitro culture of embryos isolated walnut (Juglans regia L.)Hort.Abstracts.55 (10):7533
Dennis P. Stimart, Peter D. Ascher and Harold F.Wildins. 1982.Overcoming dormancy in L. longiflorum bulblets produced in tissue culture [J].J. Amer Soc. Hort. Sci.107(6): 1004～1007
Dong-Sheng Han, Yoshiji Niimi, Masaru Nakamo.1999.Production of doubled haploid plants through colchicine treatment of anther-derived haploid calii in the Asiatic hybrid lily‘ConnecticutKing'[J].J.Japan. Soc. Hort. Sci.68(5): 979～983
Duan YP.1997.Geminivirus resistance in transgenic tobacco erpressing mutated Bci protein[J].Molecular plant-Microbe Interaction.10:617～623
Golemboskid B,Lomonossof GP.1990.Plants transformed with a tobacco mosaic virus non-struetuctural gene sequence are resistant to the Virus.Proc Natol Acad sci USA.87:6311～6315
HoshiY , KondoM , KohayashiH . 2004 . Production of transgenic lily plants by Agrobacterium-mediated transformation.PLant Cell Report.22:359～364
Hsueh-Shih Lin,Marjo J. De Jeu , Evert Jacobsen.1998. Formation of shoots from leaf axils of Alstroemeria: The effect of the position on the stem.Plant Cell, Tissue and Organ Culture.52: 165～169
IGNAT EVA IP. 1999.Ontogenetic morphogenesis of lanceolate ceaved lily (tiger lily) grown from bulbils[J].Horticultural Abstracts.69(8): 71～75
Ivanova N.V.1981.Isolated Tissue Culture for Lily Propagation.Bynlleten, Glavnogo Botanicheskogo sada.121:87～92
Jach G,Gornhardt B.1995.Enhanced quantitative resistance against fungal disease by combinatorial expression of different barley antifungal proteins in transgenic tobacco.The plant.8(1): 97～109
Joung Hyang Young and Sung Moonsuk.1995.In vitro Propagation of L.oriental hybrid‘Casa Blanca'and L.leichtlinii var. tigrinum as influenced by growth regulators and cultural explant [J].RDA Journal of Agricultural Science Horticulture.37(1):378～383
Juan Bernardo, Pe′rez-Herna′ndez , Purificacio′n Rosell-Garc?′a . 2008 . Inflorescence proliferation for somatic embryogenesis induction and suspension-derived plant regeneration from banana (Musa AAA, cv.‘Dwarf Cavendish’) male flowers.Plant Cell Rep.
KeiieHi Okazaki, Yoshito Asano, Katsuji Oosawa.1994.Interspecific hybrids between Lilium Oriental hybrid and L. Asiatic hybrid produced by embryo culture with revised midia.Breeding science.44:59～64
KI HYUN RYU,HYE WON PANK,JANG KYUNG CHOI.2002.Characterization and sequenee analysis of a lily isolate of cucumber mosaic virus form lilium tsingtauense.Journal Plant Patho1.18(2):85～92
LANGEVELD S A,GERRITS M M,DERKS A F L M ,BOONEKAMP P M,BOL J F.1995.Transformation of lily by Agrobacterium.Euphytica.85:97～100
Li. J., Brader. G.,Palva. E.T.2004.The WRKY70 transcription factor: a node of convergence for jasmonate-mediated and salicylate-mediated signals in plant defense.Plant Cell.16:319～331
Lodge J K.1993.Broad-spectrum virus resistance in transgenic plants expressing pokeweed antiviral protein.Pro Natl Acad Sci USA.90:7089～7093
Logemann J,Jach G,Tommerup H. 1992.Expression of a barley ribosome- inactivating protein leads to increased fungal protection in transgenic tobacco plants.Bio/Technology.10: 305～308
Loretta Bacchetta , Patrizio C. Remotti , Claudia Bernardini & Francesco Saccardo. 2003. Adventitious shoot regeneration from leaf explants and stem nodes of Lilium.Plant Cell, Tissue and Organ Culture.74: 37～44
Makanda.R.,Essig. J.S.,Schapaugh. M. A.,Trick. H. N.,Shah. J.2006.Genetically Engineered Resistance to Fusarium Head Blight in Wheat by Expression of Arabidopsis NPR1. Mol plant Microbe Interact.19:123～129
Mei-Kuen Yip,Hsiang-En Huang,Mang-Jye Ger,Shih-Hua Chiu,Yuh-Chih Tsai,Chin-I Lin,Teng-Yung Feng.2007.Production of soft rot resistant calla lily by expressing a ferredoxin-like protein gene (pflp) in transgenic plants[J].Plant Cell Rep.26:449～457
Merel M. Langens-Gerrits,William B.M. Miller,Anton F. Croes,Geert-Jan de Klerk. 2003.Effect of low temperature on dormancy breaking and growth after planting in lily bulblets regenerated in vitro.Plant Growth Regulation.40: 267～275
MIYOSHI H,USAMI T,TANAKA I.1995.High level of GUS gene expression driven by pollen-specific promoters in electroporated lily pollen protoplasts.Sex Plant Rep.8(4):205～209
M. Ogaki,Y. Furuichi,K. Kuroda,D. P. Chin,Y.Ogawa.2008.Importance of co-cultivation medium pH for successful Agrobacterium-mediated transformation of Lilium×formolongi.Plant Cell Rep.27:699～705
Nadel B L, Altman A, liv M.1990.Regulation of Somatic Embryogenesis in Celery Cell Suspension Early delection of Embryogenic Potential and the Induction of Synchronized Cell Culture.Plant Cell Tissue and Organ Culture. 20:119～124
Nelson R. 1993.Tobacco mosaic virus infection of transgenic N icotiana plants is inhibited by antisense constructs directed at the 5'region of viral-RNA Gene. 127:227～232
NHUT D T.2001.Shoot induction and plant regenration from reeptacle tissues of Lilium longiflorum.Sci Hort.87:131～138
Niimi.2000.In vitro bulblet formation from leaf segment of lilies especially L. rubellum Baket.Journal of the Japanese Society for Hor.Sei.69(2): 161～165
Powell Abel p.1989.Protection against tobacco mosaic virus in transgenic plants that express tobacco mosaic virus antisense RNA.Proc Natl A cad Sci VSA.86:6949～6952
Production of transgenic lily plants by Agrobacterium-mediated transformation.Plant Cell Rep.22:359～364
Robb S M. 1957. The culture of excised tissue from bulb scales of lilium specioum Thun. J Exp Bot.8:348～352
S. KUMAR,M. KASHYAP ,D.R. SHARMA.2005.In vitro regeneration and bulblet growth from lily bulbscale explants as affected by retardants, sucrose and irradiance.BIOLOGIA PLANTARUM.49 (4): 629～632
Satoch S, Kamada H,Harada H.1986.Auxin Controlled Glycoprotein Relase Into the Medium of Embroygenetic Carrot cells.Plant Physiol.81:931～993
Scaramazzi F et al.1988. Comples Rendudes Seainces de la Societe de Biologic et de ses Fililaes.France.182(2): 186～195
Su-Sung Kim,Dong-II Shin,Hee-Sung Park.2007. Transientβ-glucuronidase expression in lily (Lilium longflorum L.) pollen via wounding-assisted Agrobacterium-mediated transformation.Biotechnol Lett.29:965～969
Takayama S and Misawa M. 1979.Differentiation in Lilium bulb grown in vitro, effect of various cultural conditions.Physiologia Plantarum.46(2): 184～190
Takayama S and Misawa M. 1983.A Scheme for mass propagation of Lilium in vitro.Scientia Horticulturae.18(4): 353～362
Takayma S et al.1980.Differentiation in Lilium bulbs scales grown in vitro, Effects of Activated charoal, Physiological get of bulbs and sucrose concentation on differentiation and scale leaf formation in vitro. Physiol plant.48: 121～125
Toshikazu Matsumoto.1996.Cryopveservation of in vitro-grow apical meristems of lily (Lilium L.) by encapsulation vitrification method.Plant tissue culture letters.13(1):34
TOSHINORI T,MASAHIRO N,MOTOAKI S,ATSUSHI S,KUNISUKE T,KOHE II,HIROMICHI M.1995.Successful expression in pollen of various plants pecies of in vitro synthesized mRNA introduced by particle bombardment[J].Plant Molecular Biology.28(2):337
Van TUYL J M.Van DIEN M P,Van CREIJ M G M,Van KLEINWEE T C M,FRANKENJ,BINO R J.1991.Application of in vitro pollination,ovary culture.ovule culture and embryo rescue for overcoming incongruity barriers in interspecific Liliumc rosses.Plant Science.74:115～126
WATAD A A,YUN D J,MATSUMOTO T,NIU X,WU Y,KONONOWICZ A K,BRESSAN R A,HASEGAWA P M.1998.Microprojectile bombardment-mediated transformation of Lilium longiflorum.Plant Cell Reports.17:262～267
Wisnienskil A,Powell PA.1990.Localand systemil spread of tobacco mosaic virus in transgenic tobacco plants.The plant cell.2:559～567
Woloshuk C P.1991.Pathogen-induced proteins with inhibitory activity toward Phytophthora infestans.Plant cell.3(6): 619～628
WU F S,FENG T Y .1999.Delivery of plasmid DNA into intact plant cells by electroporation of plasmolyzed cells.Plant Cell Rep.18(5):381～386
Y. Hoshi,M. Kondo,S. Mori,Y. Adachi,M. Nakano,H. Kobayashi.2004.Production of transgenic lily plants by Agrobacterium-mediated transformation.Plant Cell Rep.22:359～364
Yoshito Asano.1980.Studies on crosses between distantly related species of Lilies V. characteristics of newly obtained hybrids through embryo culture.J. Japan. Soc. Hort. Sci.49(2): 241～250
曾镭,刘燕.2007.植物组织培养中褐化问题的研究进展.安徽农学通报.13(14):49～50
陈秋萍.2002.福建省百合病害调查初报.福建林学院学报.20(2):162～164
陈学林,李卫锋,黄海涛.2007.荷兰百合索尔邦的离体组织培养.安徽农业科学.35(20):6045～6046
陈以峰.1996.植物体细胞胚胎发生的诱导与调节研究进展.生物学杂志.69(1):3～6
程爱芳.2006.百合(Lilium spp)的组培快速繁殖技术研究.武汉生物工程学院学报.2(1):27-29
程金水.2001.园林植物遗传育种学.中国林业出版社.309～312
程佑发,王勋陵.2001.枣树体细胞胚发生的组织学研究.西北植物学报.211: 142～145
褚云霞,张永春,杨红娟等.2002.百合花药愈伤组织诱导中的基因型差异.上海农业学报.18(3):17～20
邓晓玲,孔维文,冯震,郭明,李华平. 2004.甘蓝型油菜中NPR1基因的克隆、载体构建及其转化的研究.2004年中国植物病理学会年会论文集:570～571
丁兰,刘国安,田卫东. 2005.新铁炮百合组织培养和快速繁殖研究.西北师范大学学报.37(1): 80～82
范慈惠. 1981.不同光源对百合珠芽培养的影响.江苏农业科学.3:205～207
方华舟,洪万泓.2006.百合组织培养及快繁技术的研究与探讨.贵州林业科学.34(4): 27～29
郝建平,金竹萍,王永康等.2006.枣合子胚和体细胞胚发育过程的观察与比较.分子细胞生物学报.39(5):423～426
贺晓庆.2007.NPKl和NPR1基因的克隆及其转化辣椒的研究.西北农林科技大学硕士论文
胡凤仙. 2003.转拟南芥系统获得抗性调节基因(NPRl)烟草的获得及其遗传分析.广西大学硕十学位论文
胡风荣,席梦利,刘光欣,池坚.东方百合的器官发生与体胚发生的研究.南京林业大学学报(自然科学版).31(2):5～8
黄济明,赵晓艺,张国民等. 1990.玫红百合为亲本育成百合种间杂种.园艺学报.17 (2):153～156
黄济明. 1982.王百合×大卫百合种间远源杂种的育成.园艺学报.9(3):51～56
郎楷永. 2003.中国的百合.科技导报.7:24,37～40
李铁军,郁伟. 2002.百合主要病害的识别与防治.农业科技通讯.(1):30～31
李小军. 2005.农杆菌介导NPR1基因转化安祖花的研究.上海师范大学硕士论文
梁巧兰,徐秉良.2004.观赏百合根腐病病原鉴定及药剂筛选.甘肃农业大学学报.39(1):25～28
刘芬,王发林. 2001.兰州百合花丝组培诱导完整植株的研究.甘肃农业科技.6: 29～31
刘何,谢令琴,赵建军.2007.植物抗性相关基因的研究.华北农学报.22:17～20
刘菊华,金志强,徐碧玉等.2003.龙牙百合的遗传转化及再生.分子植物育种. 1(4):465～474
刘明志,林雪艳. 2002.激素对百合植株再生的影响.广西植物.22(2): 167～170
刘选明,周朴华,屈妹存等.1997.百合鳞片叶离体诱导形成不定芽和体细胞胚.园艺学报.24(4):353～358
卢其能. 2002.龙牙百合的组织培养和快速繁殖研究.江西农业学报.14(4): 43～46
陆家云.1995.药用植物病害.中国农业出版社.45～68
罗荡平. 2004.白叶枯病和稻瘟病拟南芥NPR1基因转入水稻使水稻获得广谱抗病性.广西大学硕士学位论文
罗凤霞,徐桂华,金丽丽等. 2000.新铁炮百合微繁的研究.沈阳农业大学学报.31(3): 254～257
罗其德. 2004.Genetic Modification to Enhance Tomato Resistance to Disease.广西大学硕士论文
毛元荣. 2004.洋桔梗叶片再生体系的建立及农杆菌介导的NPR1基因转化洋桔梗的初步研究.上海师范大学硕士论文
莫昭展,符韵林,戚萌. 2007.野百合鳞茎芽的诱导和增殖的初步研究.安徽农业科学. 35(31):9890～9892
欧阳秩,吴帮承.1996.观赏植物病害.中国农业出版社.23～65
乔永旭,陈超,张永平等.2007.东方百合“索邦”离体培养再生体系的建立.安徽农业科学.35(1):67～69
沈淑琳.1996.百合病毒及其检验.植物检疫.10(4):223～225
衰澍,贾勇炯,林宏辉. 2003.诱导植物体细胞胚发生的几个生理因素.植物生理学通讯.39(5):508～512.
唐东芹,钱虹妹,唐克轩等.2004.根癌农杆菌介导半夏凝集素基因pBIXPTA对百合的转化.上海交通大学学报.22(2):135～137
唐东芹,钱虹妹等. 2003.百合基因转化胚性愈伤组织受体系统的建立.浙江林学院学报.20(3):273～276
唐祥宁,游春平,刘福秀等.1997.江西百合病害调查与鉴定.江西农业学报.9(4):l～8
唐益苗. 2006.小麦及其近缘植物NPR1类似基因和反转录转座子基因片段的分离和特性分析.中国农业科学院博士论文
汪海洋,张慧珍,李炳武等.2002.百合病害的发生与防治.上海蔬菜.4:35～36
王爱勤,周歧伟,何龙飞等.1998.百合试管结鳞茎的研究.广西农业大学学报.17(1):71～75
王刚,杜捷,李桂英等. 2002.兰州百合和野百合组织培养及快速繁殖研究.西北师范大学学报(自然科学版).38(1): 69～71
王关林,方宏筠等. 2002.抗菌肽基因转化大白菜获得抗病转基因植株及稳定遗传.植物学报.44(8):951～955
王季林. 1987.百合鳞片不同部位繁殖系数的观察.江苏农业科学.1: 30
王清连,王敏,师海荣. 2005.植物激素对棉花体细胞胚胎发生的诱导及调节作用.生物技术通讯.15(6):577～579
王仁祥.2005.转基因植物研究进展.生物学教学.30(13):9～12
温春秀,及华. 1997.新铁炮百合((L. longiflorum×L. formosanum)的离体快速繁殖.植物生理学通讯.33(3): 198
吴乃虎编著. 2002.基因工程原理(第二版下册).北京:科学出版社.171～225.
吴若普,阮少宁等. 2003.百合胚状体形成途径及植株的再生研究.福建林学院学报. 23(4):322～325
徐碧玉,刘菊华,金志强.2005.类黄酮3',5'羟基化酶基因的克隆及转化铁炮百合.园艺学报.
许宝辉. 2002.百合子房组织培养研究.南科技大学学报.3:35～38
杨乃博. 1981.十二种植物的试管无性繁殖.植物学报.23(4):284
姚祥坦,曹家树,李晋豫,王神云.2004.植物抗病毒病育种策略.细胞生物学杂志.26:362～366
叶世森,林芳,宋建英. 2005.百合病害的研究综述.西南林学院学报.25(3):84～88
于湄,叶长明. 1999.转基因植物中RNA介导病毒抗性.植物学通报.16(2) :127～130
张德水,陈受宜. 1997.植物抗病性的分子生物学研究进展.植物病理学报.27(2):97～103
张丽珍.2005.NPR1基因的克隆及西瓜的遗传转化的研究.广西师范大学硕士论文.
张兴翠,周昌华,殷家明等. 2003.药用百合的多倍体诱导及快速繁殖.西南农业大学学报.25(1):14～17
张艳贞,魏松红,杜娟,王罡. 2000.植物抗病分子机制及抗病基因工程研究进展.沈阳农业大学学报.31(4):365～369
张艺萍,屈云慧,王祥宁等.2007.提高百合茎尖组织培养脱毒效率研究.广东农业科学.1:37～38
张于光,肖启明.2003.百合的主要病害及其防治.植物杂志.(3):16～17
张云,原雅玲,刘青林. 2001.百合品种改良与生物技术研究进展.北京林业大学学报. 23(6): 56～59
张中义. 1992.观赏植物真菌病害.四川科技出版社.26～57
赵庆芳,曾小英,丁兰等. 2003.东方百合组织培养和快速繁殖研究.西北师范大学学报(自然科学版).39(1):66～68
张玉静,王志刚,印东生,王平.2007.百合主要病害及防治.北方园艺.11:202
张志兰.2007.山茶花组织培养过程忠的防外植体褐化试验.西部林业科学.36(2):118～121
郑秀芳,谬为. 2002.抗菌肤及在植物抗病基因工程中的应用.生物学通报.37(10): 9～10
钟景辉,蔡秋锦,张飞萍等.2000.百合病害及其持续治理.森林病虫通讯.19(2):28～31,23.
赵欢蕊,刘雅莉,王跃进.2007.农杆菌介导的普利安娜百合鳞茎直接分化受体系统的建立.西北农业学报.16(4):142～147
周俊彦.1981.植物体细胞在组织培养中产生的胚状体I植物体细胞的胚状体发生.植物生理学报.7(4):391～395
周晓燕. 2002.百合主要病害及其防治.植物保护.28(1):57～58
周艳萍,郑红娟,贾桂霞. 2007.两个亚洲百合品种离体再生体系的建立.北京林业大学学报.29(1):123～127
诸葛强,王婕琛,陈英等. 2003.新疆杨高效遗传转化系统的建立.植物资源与环境学报. 12(4):6～10
邹新慧,何平.2002.植物抗病毒基因工程研究进展及存在的问题.江西农业报. 14(4):59～65
A. A. Watad,D.-J. Yun,T. Matsumoto,X. Niu,Y. Wu,A. K. Kononowicz,R. A. Bressan,P. M. Hasegawa . 1998 . Microprojectile bombardment-mediated transformation of Lilium longiflorum.Plant Cell Reports.17: 262～267
Adam M,Dimech,Rob Cross,Rebecca Ford.2007.Micropropagation of Gymea Lily (Doryanthes excelsa Correa) from New South Wales, Australia.Plant Cell, Tissue and Organ Culture.88: 157～165
Akira Kikuchi, Nobuya Sanuki, Katsumi Higashi.2006.Abscisic acid and stress treatment are essential for the acquisition of embryogenic competence by carrot somatic.Planta.223:637~645
Arzate F A M,Nakazaki T,Okumoto Y et al.1997.Efficient callus induction and plant regenration from filaments with anther in lily (L.longiflorum).Plant Cell Reports.16:836～840 BELLARDI M G,NANNI G.2002.Old and new viruses of lily in Italy.Acta Horticulturae.568:215～220
Cao. H.,Li. X.,Dong .X.1998.Generation of broad-spectrum disease resistance by over expression of an essential regulatory gene in systemic acquired resistance.Proc Natl Acad Sci USA.95:6531～6536
Carr JP,Gal DNA et al.1994.Replicase-mediated resistance to cucumber mosaic virus in transgenic plants involves suppression of both virus replication in the inoculated leaves and long distance move meat.Virology.199:439～447
Chern. M., Canlas.P.E., Fitzgerald. H. A., Ronald. P. C. 2005.Rice NRR, a negative regulator of disease resistance,interacts with Arabidopsis NPR1 and rice NH1.Plant Journal.43: 623～35
Chern. M.,Fitzgerald. H.A.,Canlas. P. E.,Navarre. D. A.,Ronald. P. C.2005. Over expression of a rice NPR1 homolog leads to constitutive activation of defense response and hypersensitivity to light. Mol Plant Microbe Interact.18: 511～520
Chern. M.S.,Fitzgerald. H.A.,Yadav. R.C.,Canlas. P. E..et al.2001.Evidence for a disease-resistance pathway in rice similar to the NPR1-mediated signaling pathway in Arabidopsis.Plant Journal.27:101～113
COHEN A,MEREDITHC P,SANIEWSKIM et al .1992.A grobacterium-mediated transformation of Lilium[J].ActaH ort.325:611～618
COHEN J,GERA A.1996.Virus diseases of lilies in Israel[J].Acta Horticulture.432:84～87
Cooper B,Lapidot M et al.1995.A defective movement protein of TMV in transgenic plants confers resistant to multiple viruses whereas the functional analog in creases susceptibility[J]. Virology.206:307～313
Cossio F. et al.1985.Further findings on the in vitro culture of embryos isolated walnut (Juglans regia L.)Hort.Abstracts.55 (10):7533
Dennis P. Stimart, Peter D. Ascher and Harold F.Wildins. 1982.Overcoming dormancy in L. longiflorum bulblets produced in tissue culture [J].J. Amer Soc. Hort. Sci.107(6): 1004～1007
Dong-Sheng Han, Yoshiji Niimi, Masaru Nakamo.1999.Production of doubled haploid plants through colchicine treatment of anther-derived haploid calii in the Asiatic hybrid lily‘ConnecticutKing'[J].J.Japan. Soc. Hort. Sci.68(5): 979～983
Duan YP.1997.Geminivirus resistance in transgenic tobacco erpressing mutated Bci protein[J].Molecular plant-Microbe Interaction.10:617～623
Golemboskid B,Lomonossof GP.1990.Plants transformed with a tobacco mosaic virus non-struetuctural gene sequence are resistant to the Virus.Proc Natol Acad sci USA.87:6311～6315
HoshiY , KondoM , KohayashiH . 2004 . Production of transgenic lily plants by Agrobacterium-mediated transformation.PLant Cell Report.22:359～364
Hsueh-Shih Lin,Marjo J. De Jeu , Evert Jacobsen.1998. Formation of shoots from leaf axils of Alstroemeria: The effect of the position on the stem.Plant Cell, Tissue and Organ Culture.52: 165～169
IGNAT EVA IP. 1999.Ontogenetic morphogenesis of lanceolate ceaved lily (tiger lily) grown from bulbils[J].Horticultural Abstracts.69(8): 71～75
Ivanova N.V.1981.Isolated Tissue Culture for Lily Propagation.Bynlleten, Glavnogo Botanicheskogo sada.121:87～92
Jach G,Gornhardt B.1995.Enhanced quantitative resistance against fungal disease by combinatorial expression of different barley antifungal proteins in transgenic tobacco.The plant.8(1): 97～109
Joung Hyang Young and Sung Moonsuk.1995.In vitro Propagation of L.oriental hybrid‘Casa Blanca'and L.leichtlinii var. tigrinum as influenced by growth regulators and cultural explant [J].RDA Journal of Agricultural Science Horticulture.37(1):378～383
Juan Bernardo, Pe′rez-Herna′ndez , Purificacio′n Rosell-Garc?′a . 2008 . Inflorescence proliferation for somatic embryogenesis induction and suspension-derived plant regeneration from banana (Musa AAA, cv.‘Dwarf Cavendish’) male flowers.Plant Cell Rep.
KeiieHi Okazaki, Yoshito Asano, Katsuji Oosawa.1994.Interspecific hybrids between Lilium Oriental hybrid and L. Asiatic hybrid produced by embryo culture with revised midia.Breeding science.44:59～64
KI HYUN RYU,HYE WON PANK,JANG KYUNG CHOI.2002.Characterization and sequenee analysis of a lily isolate of cucumber mosaic virus form lilium tsingtauense.Journal Plant Patho1.18(2):85～92
LANGEVELD S A,GERRITS M M,DERKS A F L M ,BOONEKAMP P M,BOL J F.1995.Transformation of lily by Agrobacterium.Euphytica.85:97～100
Li. J., Brader. G.,Palva. E.T.2004.The WRKY70 transcription factor: a node of convergence for jasmonate-mediated and salicylate-mediated signals in plant defense.Plant Cell.16:319～331
Lodge J K.1993.Broad-spectrum virus resistance in transgenic plants expressing pokeweed antiviral protein.Pro Natl Acad Sci USA.90:7089～7093
Logemann J,Jach G,Tommerup H. 1992.Expression of a barley ribosome- inactivating protein leads to increased fungal protection in transgenic tobacco plants.Bio/Technology.10: 305～308
Loretta Bacchetta , Patrizio C. Remotti , Claudia Bernardini & Francesco Saccardo. 2003. Adventitious shoot regeneration from leaf explants and stem nodes of Lilium.Plant Cell, Tissue and Organ Culture.74: 37～44
Makanda.R.,Essig. J.S.,Schapaugh. M. A.,Trick. H. N.,Shah. J.2006.Genetically Engineered Resistance to Fusarium Head Blight in Wheat by Expression of Arabidopsis NPR1. Mol plant Microbe Interact.19:123～129
Mei-Kuen Yip,Hsiang-En Huang,Mang-Jye Ger,Shih-Hua Chiu,Yuh-Chih Tsai,Chin-I Lin,Teng-Yung Feng.2007.Production of soft rot resistant calla lily by expressing a ferredoxin-like protein gene (pflp) in transgenic plants[J].Plant Cell Rep.26:449～457
Merel M. Langens-Gerrits,William B.M. Miller,Anton F. Croes,Geert-Jan de Klerk. 2003.Effect of low temperature on dormancy breaking and growth after planting in lily bulblets regenerated in vitro.Plant Growth Regulation.40: 267～275
MIYOSHI H,USAMI T,TANAKA I.1995.High level of GUS gene expression driven by pollen-specific promoters in electroporated lily pollen protoplasts.Sex Plant Rep.8(4):205～209
M. Ogaki,Y. Furuichi,K. Kuroda,D. P. Chin,Y.Ogawa.2008.Importance of co-cultivation medium pH for successful Agrobacterium-mediated transformation of Lilium×formolongi.Plant Cell Rep.27:699～705
Nadel B L, Altman A, liv M.1990.Regulation of Somatic Embryogenesis in Celery Cell Suspension Early delection of Embryogenic Potential and the Induction of Synchronized Cell Culture.Plant Cell Tissue and Organ Culture. 20:119～124
Nelson R. 1993.Tobacco mosaic virus infection of transgenic N icotiana plants is inhibited by antisense constructs directed at the 5'region of viral-RNA Gene. 127:227～232
NHUT D T.2001.Shoot induction and plant regenration from reeptacle tissues of Lilium longiflorum.Sci Hort.87:131～138
Niimi.2000.In vitro bulblet formation from leaf segment of lilies especially L. rubellum Baket.Journal of the Japanese Society for Hor.Sei.69(2): 161～165
Powell Abel p.1989.Protection against tobacco mosaic virus in transgenic plants that express tobacco mosaic virus antisense RNA.Proc Natl A cad Sci VSA.86:6949～6952
Production of transgenic lily plants by Agrobacterium-mediated transformation.Plant Cell Rep.22:359～364
Robb S M. 1957. The culture of excised tissue from bulb scales of lilium specioum Thun. J Exp Bot.8:348～352
S. KUMAR,M. KASHYAP ,D.R. SHARMA.2005.In vitro regeneration and bulblet growth from lily bulbscale explants as affected by retardants, sucrose and irradiance.BIOLOGIA PLANTARUM.49 (4): 629～632
Satoch S, Kamada H,Harada H.1986.Auxin Controlled Glycoprotein Relase Into the Medium of Embroygenetic Carrot cells.Plant Physiol.81:931～993
Scaramazzi F et al.1988. Comples Rendudes Seainces de la Societe de Biologic et de ses Fililaes.France.182(2): 186～195
Su-Sung Kim,Dong-II Shin,Hee-Sung Park.2007. Transientβ-glucuronidase expression in lily (Lilium longflorum L.) pollen via wounding-assisted Agrobacterium-mediated transformation.Biotechnol Lett.29:965～969
Takayama S and Misawa M. 1979.Differentiation in Lilium bulb grown in vitro, effect of various cultural conditions.Physiologia Plantarum.46(2): 184～190
Takayama S and Misawa M. 1983.A Scheme for mass propagation of Lilium in vitro.Scientia Horticulturae.18(4): 353～362
Takayma S et al.1980.Differentiation in Lilium bulbs scales grown in vitro, Effects of Activated charoal, Physiological get of bulbs and sucrose concentation on differentiation and scale leaf formation in vitro. Physiol plant.48: 121～125
Toshikazu Matsumoto.1996.Cryopveservation of in vitro-grow apical meristems of lily (Lilium L.) by encapsulation vitrification method.Plant tissue culture letters.13(1):34
TOSHINORI T,MASAHIRO N,MOTOAKI S,ATSUSHI S,KUNISUKE T,KOHE II,HIROMICHI M.1995.Successful expression in pollen of various plants pecies of in vitro synthesized mRNA introduced by particle bombardment[J].Plant Molecular Biology.28(2):337
Van TUYL J M.Van DIEN M P,Van CREIJ M G M,Van KLEINWEE T C M,FRANKENJ,BINO R J.1991.Application of in vitro pollination,ovary culture.ovule culture and embryo rescue for overcoming incongruity barriers in interspecific Liliumc rosses.Plant Science.74:115～126
WATAD A A,YUN D J,MATSUMOTO T,NIU X,WU Y,KONONOWICZ A K,BRESSAN R A,HASEGAWA P M.1998.Microprojectile bombardment-mediated transformation of Lilium longiflorum.Plant Cell Reports.17:262～267
Wisnienskil A,Powell PA.1990.Localand systemil spread of tobacco mosaic virus in transgenic tobacco plants.The plant cell.2:559～567
Woloshuk C P.1991.Pathogen-induced proteins with inhibitory activity toward Phytophthora infestans.Plant cell.3(6): 619～628
WU F S,FENG T Y .1999.Delivery of plasmid DNA into intact plant cells by electroporation of plasmolyzed cells.Plant Cell Rep.18(5):381～386
Y. Hoshi,M. Kondo,S. Mori,Y. Adachi,M. Nakano,H. Kobayashi.2004.Production of transgenic lily plants by Agrobacterium-mediated transformation.Plant Cell Rep.22:359～364
Yoshito Asano.1980.Studies on crosses between distantly related species of Lilies V. characteristics of newly obtained hybrids through embryo culture.J. Japan. Soc. Hort. Sci.49(2): 241～250