苦瓜花药培养及培养过程中内源激素研究
|
摘要
苦瓜为葫芦科苦瓜属植物,它不仅是我国及东南亚各国人民喜爱的传统蔬菜,也是一种重要的药用植物。花药培养是获得单倍体植株最有效的方法之一,对植物育种有着非常重要的意义。本试验对影响苦瓜花药培养的诸因素进行了较系统的研究,首次从苦瓜花药愈伤组织分化得到不定芽。另外,针对苦瓜花药培养易于产生愈伤组织而难以再生不定芽的问题,首次对苦瓜花药和愈伤组织的内源激素含量进行了研究。
主要的研究结果如下：
(1)苦瓜小孢子发育时期与花蕾长度,花药颜色密切相关。应当以花蕾长度和花药颜色双重标准衡量小孢子的发育时期。小孢子处于单核后期时,绿脆纵横径最小,纵径为2.597 mm,横径为1.933 mm；其他三个品种花蕾纵径在4.024-4.213 mm,横径在1.998-2.050 mm。花药颜色为青白至微黄也是小孢子发育到单核后期的特征。
(2)4℃低温预处理对苦瓜花药愈伤组织诱导率的影响因品种而异,经冷处理后,长白诱导率下降,碧秀、大白和胖妞的诱导率都显著提高。总体而言,苦瓜花药冷处理时间不宜过长,超过4天会显著降低诱导率,其最适冷处理时间为1-2天。
(3)当MS培养基中2,4-D浓度为0.5 mg/L,6-BA浓度2.0 mg/L时,各品种的花药愈伤组织诱导率最高,其中碧秀苦瓜达到了79.42%。2,4-D浓度的变化对花药愈伤组织的诱导影响明显,低浓度的2,4-D(0.1 mg/L)与BA各水平组合,都不能很好地让苦瓜花药长出愈伤组织,2,4-D浓度过高又会造成愈伤组织膨大迅速,褐化死亡也加快。
(4)苦瓜花药愈伤组织的诱导率随蔗糖浓度升高而显著降低,较高浓度的蔗糖可以有效抑制花药壁等体细胞发生愈伤组织,为了保证花药诱导率的同时也保证愈伤组织来源,培养基中添加50 g/L蔗糖最为适宜。
(5)培养温度和光周期对苦瓜花药愈伤组织诱导有着重要影响。黑暗条件下比较容易产生愈伤组织,但是愈伤组织的质地很疏松,颜色发白；光照条件下产生愈伤组织比较慢,但质地致密,颜色绿。温度越高,花药愈伤组织诱导率越高,但质地更为疏松。25℃暗培养1周再转至光培养得到的诱导率最理想。
(6)诱导阶段得到的愈伤组织按照外观和质地大致可分为两类：一类为黄白色、疏松状愈伤组织；另一类呈黄绿色、光滑颗粒状。
(7)在TDZ 0.5 mg/L和2,4-D 0.1 mg/L的组合中添加三十烷醇2.0 mg/L,碧秀苦瓜部分黄绿色愈伤组织出现深绿色芽点并最终形成不定芽,这是首次在苦瓜花药培养中分化出不定芽。在激素组合为谷氨酰胺0.1 mg/L+TDZ 0.5 mg/L+2,4-D 0.1 mg/L的培养基中,黄白色愈伤组织出现3 cm左右的竖状突起。在激素组合6-BA 0.5 mg/L+NAA 1.0 mg/L中,长白苦瓜的部分黄绿色愈伤组织分化出了根。
(8)在本试验中,长白苦瓜分化出不定根33条,根尖细胞染色体观察表明,其中4条为单倍体根,单倍体分化率为12.12%,其余为混倍体,其中三倍体较多。
(9)应用高效液相色谱法(HPLC)研究了苦瓜外植体和愈伤组织内源激素含量,结果表明：苦瓜易于诱导愈伤组织与其内源IAA含量较高关系密切；高含量ZT对于不定芽的分化非常重要；高含量的GA3会抑制不定芽的发生,低水平的GA3/ZT值对于不定芽分化至关重要。
Bitter melon (Momordica charantia L.) is not only a valuable vegetable crop but also an important medicinal plant in China and East Asia. Anther culture is one of the most effective ways of obtaining haploid and homozygotic plants, and it has special significance for the genetic improvement of plants. In this study, four cultivars of bitter melon were used for anther culture, and it was first time to differentiate buds in anther culture of bitter melon. Because explants of bitter melon were apt to form callus rather than adventitious buds, the present investigation measured and correlated endogenous hormone status on explants and calluses with their competence. The main results were as follows.
(1) The relationship between microspore developmental stages and length of flower buds, color of anthers was established. When the microspores developed at the late-uninucleate stage, the flower buds were 2.597-4.213 mm in longitudinal diameter, 1.933-2.050 mm in diameter and the anthers were green-white or light green.
(2) After 4℃cold pretreatment, callus induction rate of 3 genotypes increased while Changbai decreased. Cold pretreatment to anthers for 1-2 days resulted in the highest callus formation rate. While anthers pretreated more than 4 days, callus could hardly be induced.
(3) The callus formation rate of bitter melon reached the highest when medium added with 2,4-D 0.5mg/L and BA 2.0 mg/L, and Bixiu reached 79.42%. Low concentration of 2, 4-D (0.1 mg/L) in combination with each level of 6-BA was unable to induce callus. However, with the concentration of 2,4-D up to 1.0 mg/L, the callus was prone to brown.
(4) The induction rate decreased when the concentration of sucrose increased. Higher concentration of sucrose could inhibited callus derived from anther wall.5% sucrose in medium caused a relatively higher induction rate.
(5) Anthers cultured under illumination condition resulted in lower induction rate than those cultured in darkness, however callus under light were denser and more vigorous. The anthers cultured at 25℃in the dark for 7 days and then cultured under light resulted in the most high induction rate.
(6) Callus could be morphologically divided into two types:one type was slight yellow, loosened callus. The other type was viridescence, granular, smooth callus.
(7) On MS medium supplemented with TDZ 0.5 mg/L,2,4-D 0.1 mg/L and triacontanol 2.0 mg/L, adventitious buds differentiated from callus of Bixiu. On MS medium supplemented with glutamine 0.1 mg/L, TDZ 0.5 mg/L and 2,4-D 0.1 mg/L, protuberant structures were produced on the surface of callus. Callus of Changbai differentiated roots when medium supplemented with 6-BA 0.5 mg/L and NAA 1.0 mg/L.
(8) Anthers of Changbai differentiated 33 roots, and chromosomal examination of root tip cells showed that 4 roots were haploids.
(9) The endogenous hormone concentrations of the initial explants and calluses were determined by means of high pressure liquid chromatography (HPLC). The endogenous IAA was higher in the explants that were easier to induce callus, and higher concentrations of ZT was positive to buds differentiation, while GA3 was negative to the buds formation.
主要的研究结果如下：
(1)苦瓜小孢子发育时期与花蕾长度,花药颜色密切相关。应当以花蕾长度和花药颜色双重标准衡量小孢子的发育时期。小孢子处于单核后期时,绿脆纵横径最小,纵径为2.597 mm,横径为1.933 mm；其他三个品种花蕾纵径在4.024-4.213 mm,横径在1.998-2.050 mm。花药颜色为青白至微黄也是小孢子发育到单核后期的特征。
(2)4℃低温预处理对苦瓜花药愈伤组织诱导率的影响因品种而异,经冷处理后,长白诱导率下降,碧秀、大白和胖妞的诱导率都显著提高。总体而言,苦瓜花药冷处理时间不宜过长,超过4天会显著降低诱导率,其最适冷处理时间为1-2天。
(3)当MS培养基中2,4-D浓度为0.5 mg/L,6-BA浓度2.0 mg/L时,各品种的花药愈伤组织诱导率最高,其中碧秀苦瓜达到了79.42%。2,4-D浓度的变化对花药愈伤组织的诱导影响明显,低浓度的2,4-D(0.1 mg/L)与BA各水平组合,都不能很好地让苦瓜花药长出愈伤组织,2,4-D浓度过高又会造成愈伤组织膨大迅速,褐化死亡也加快。
(4)苦瓜花药愈伤组织的诱导率随蔗糖浓度升高而显著降低,较高浓度的蔗糖可以有效抑制花药壁等体细胞发生愈伤组织,为了保证花药诱导率的同时也保证愈伤组织来源,培养基中添加50 g/L蔗糖最为适宜。
(5)培养温度和光周期对苦瓜花药愈伤组织诱导有着重要影响。黑暗条件下比较容易产生愈伤组织,但是愈伤组织的质地很疏松,颜色发白；光照条件下产生愈伤组织比较慢,但质地致密,颜色绿。温度越高,花药愈伤组织诱导率越高,但质地更为疏松。25℃暗培养1周再转至光培养得到的诱导率最理想。
(6)诱导阶段得到的愈伤组织按照外观和质地大致可分为两类：一类为黄白色、疏松状愈伤组织；另一类呈黄绿色、光滑颗粒状。
(7)在TDZ 0.5 mg/L和2,4-D 0.1 mg/L的组合中添加三十烷醇2.0 mg/L,碧秀苦瓜部分黄绿色愈伤组织出现深绿色芽点并最终形成不定芽,这是首次在苦瓜花药培养中分化出不定芽。在激素组合为谷氨酰胺0.1 mg/L+TDZ 0.5 mg/L+2,4-D 0.1 mg/L的培养基中,黄白色愈伤组织出现3 cm左右的竖状突起。在激素组合6-BA 0.5 mg/L+NAA 1.0 mg/L中,长白苦瓜的部分黄绿色愈伤组织分化出了根。
(8)在本试验中,长白苦瓜分化出不定根33条,根尖细胞染色体观察表明,其中4条为单倍体根,单倍体分化率为12.12%,其余为混倍体,其中三倍体较多。
(9)应用高效液相色谱法(HPLC)研究了苦瓜外植体和愈伤组织内源激素含量,结果表明：苦瓜易于诱导愈伤组织与其内源IAA含量较高关系密切；高含量ZT对于不定芽的分化非常重要；高含量的GA3会抑制不定芽的发生,低水平的GA3/ZT值对于不定芽分化至关重要。
Bitter melon (Momordica charantia L.) is not only a valuable vegetable crop but also an important medicinal plant in China and East Asia. Anther culture is one of the most effective ways of obtaining haploid and homozygotic plants, and it has special significance for the genetic improvement of plants. In this study, four cultivars of bitter melon were used for anther culture, and it was first time to differentiate buds in anther culture of bitter melon. Because explants of bitter melon were apt to form callus rather than adventitious buds, the present investigation measured and correlated endogenous hormone status on explants and calluses with their competence. The main results were as follows.
(1) The relationship between microspore developmental stages and length of flower buds, color of anthers was established. When the microspores developed at the late-uninucleate stage, the flower buds were 2.597-4.213 mm in longitudinal diameter, 1.933-2.050 mm in diameter and the anthers were green-white or light green.
(2) After 4℃cold pretreatment, callus induction rate of 3 genotypes increased while Changbai decreased. Cold pretreatment to anthers for 1-2 days resulted in the highest callus formation rate. While anthers pretreated more than 4 days, callus could hardly be induced.
(3) The callus formation rate of bitter melon reached the highest when medium added with 2,4-D 0.5mg/L and BA 2.0 mg/L, and Bixiu reached 79.42%. Low concentration of 2, 4-D (0.1 mg/L) in combination with each level of 6-BA was unable to induce callus. However, with the concentration of 2,4-D up to 1.0 mg/L, the callus was prone to brown.
(4) The induction rate decreased when the concentration of sucrose increased. Higher concentration of sucrose could inhibited callus derived from anther wall.5% sucrose in medium caused a relatively higher induction rate.
(5) Anthers cultured under illumination condition resulted in lower induction rate than those cultured in darkness, however callus under light were denser and more vigorous. The anthers cultured at 25℃in the dark for 7 days and then cultured under light resulted in the most high induction rate.
(6) Callus could be morphologically divided into two types:one type was slight yellow, loosened callus. The other type was viridescence, granular, smooth callus.
(7) On MS medium supplemented with TDZ 0.5 mg/L,2,4-D 0.1 mg/L and triacontanol 2.0 mg/L, adventitious buds differentiated from callus of Bixiu. On MS medium supplemented with glutamine 0.1 mg/L, TDZ 0.5 mg/L and 2,4-D 0.1 mg/L, protuberant structures were produced on the surface of callus. Callus of Changbai differentiated roots when medium supplemented with 6-BA 0.5 mg/L and NAA 1.0 mg/L.
(8) Anthers of Changbai differentiated 33 roots, and chromosomal examination of root tip cells showed that 4 roots were haploids.
(9) The endogenous hormone concentrations of the initial explants and calluses were determined by means of high pressure liquid chromatography (HPLC). The endogenous IAA was higher in the explants that were easier to induce callus, and higher concentrations of ZT was positive to buds differentiation, while GA3 was negative to the buds formation.
引文
曹家树,申书兴.园艺植物育种学[M].中国农业大学出版社,2001
曹利仙,赵鹉,唐宇力等.硝酸银对黄瓜离体子叶培养芽再生的促进效应[J].甘肃农业大学学报,2001,36(2)：168-171
陈佳.苦瓜花药培养诱导愈伤组织及器官分化的研究[D].四川农业大学,2007
陈晓,詹玉丝,徐小利等.花药培养建立辣椒DH纯系的初步研究[J].河南农业科学,2003(3)：52-55
陈振光.园艺植物离体培养学[M].中国农业出版社,1995,23-25
盛艳萍,杨建平,张松等.大葱成熟胚离体再生过程中内源激素的变化[J].园艺学报.2005,32(3)：318-320
代色平,包满珠.矮牵牛花药培养及植株再生研究[J].亚热带植物科学,2003,32(2)：55-57
邓向东,耿玉轩,路子显等.外植体和培养因子对哈密瓜不定芽诱导的影响[J].园艺学报,1996,23(1)：57-61
董艳荣.甜瓜花药再生体系的基础研究[D].南京农业大学,2002
冯玉生,王则僖.紫云英植物诱导初报[J].科学通报,1981,3：30-34
付凤玲,冯质雷,渠柏艳等.玉米未成熟胚胚性愈伤组织诱导率与内源激素含量的关系[J].核农学报,2006,20(1)：10
耿晨光,何刚.瓜类花药培养中激素和碳源的应用研究进展[J].长江蔬菜,2008,7b：5-9
郭子彪,盖均镒.内源激素IAA、ABA对大豆萌发子叶胚性愈伤组织诱导及其分化的调控[J].大豆科学,1997,16(3)：194-198
韩丽华.厚皮甜瓜未受精胚珠离体培养技术[D].河北农业大学,2004
何晓明,谢大森,陈清华等.我国瓜类细胞工程研究进展[J].深圳职业技术学院学报,2002(3)：18-19
何艳.苦瓜花药离体培养初探[D].四川农业大学,2008
何艳,李焕秀,郑阳霞等.苦瓜花药培养诱导愈伤组织的研究[J].北方园艺,2008(12)：164-16
缑艳霞.西瓜花药离体培养技术研究[D].浙江大学,2006
黄荣韶,覃伟.无籽西瓜离体培养中玻璃化现象的研究[J].广西农业大学学报,1998,17(3)：266-270
黄泰康,丁志遵,赵守训等.现代本草纲目(上卷)[M].中国医药科技出版社,2000
季彪俊,江树业,陈启锋等.活性炭在水稻花药培养中的作用[J].福建农业大学学报,1998,27(1)：16-19
李焕秀,Murch S.J., Sacena P.K.. TDZ诱导的黄芩外植体再生研究[J].四川农业大学学报,2000,18(4)：325
李娟.西瓜花药和小孢子培养技术研究[D].四川农业大学,2008
李娟,张丽,李焕秀等.西瓜花药培养技术研究[J].中国瓜菜,2008(4)：4-8
李俊明.植物组织培养教程[M].北京,北京农业大学出版社,1992
李文泽,景健康,姚根怀等.基因型和甘露醇预处理对大麦花粉植株高频率再生的影响[J].科学通报,1992,10：935-938
李文泽,宋子红,景健康等.甘露醇预处理对大麦雄核发育的影响[J].植物学报,1995,37(7)：552-557
李锡香,沈镝,王海平.我国蔬菜种质资源研究进展与发展策略[J].中国蔬菜,2006(增刊)：3-9
李雪梅,刘熔山.小麦幼穗胚性愈伤组织诱导及分化过程中内源激素的作用[J].植物生理学通讯,1994,30(4)：255-260
李宗霆,周燮.植物激素及其免疫检测技术[M].南京,江苏科学技术出版社,1996
刘涤,迟静芬,刘桂芸.烟草愈伤组织器官发生过程中外源激素的作用[J].植物生理与分子生物学报,1986,12(1)：104
刘贵仁,严仁玲,张磊等.石刁柏花药培养与染色体倍性的观察[J].吉林农业大学学报,1992,12(1)：5-8
刘京贞.天竺葵的应用及组织培养[J].北方园艺,2002(2)：54-56
罗言云.麻疯树花药培养及两个核糖体失活蛋白基因的表达研究[D].四川大学,2006
马刘峰.甜瓜花药离体培养研究[D].石河子大学,2006
马刘峰等.甜瓜花药愈伤组织分化与激素处理间的关系[J].中国瓜菜,2006(2)：4-6
潘瑞炽.植物组织培养[M].广东高等教育出版社,2001
彭爱芝,李劲.苦瓜中氨基酸和无机元素的含量分析[J].湖南医科大学学报,1996,21(4)：305-307
彭新红,周健松.芦笋花药培养胚状体诱导条件优化的初探[J].江西农业大学学报,2006,28(1)：39-43
戚春章,胡是麟,漆小泉.中国蔬菜种质资源的种类及分布[J].作物品种资源,1997,(1)：1-5
宋莉英,高峰.苦瓜离体培养过程中内源激素含量的变化[J].植物学通报,2006,(2)：66-70
苏华,金宝燕,任华中.黄瓜花药培养中若干影响因素的研究[J].沈阳农业大学学报,2006,37(3)：470-472
孙兰英,卢淑雯.黄瓜组织培养研究进展[J].黑龙江农业科学,2004,(4)：25-27
汤国庆,李文安.在离体条件下拐芹形成愈伤组织和非胚性愈伤组织过程中几种内源激素的变化[J].实验生物学报,1995,28(2)：203
唐玉林,陈婉芳,周燮.烟草叶块分化根和芽过程中内源激素水平的变化[J].南京农业大学学报,1996,19(2)：12-16
陶正南.甜瓜花药培养诱导成植株[J].植物生理学通讯,1987,(5)：43
田文忠.提高籼稻愈伤组织再生频率的研究[J].遗传学报,1994,21(3)：215
魏瑛.低温预处理对西瓜花药愈伤组织诱导的影响[J].甘肃农业科技,1999,9：34-35
魏瑛,张俊莲,陈年来等.西瓜花药愈伤组织的诱导[J].甘肃农业大学学报,1998,33：127-130
王得元,何晓明,王鸣.蔬菜生物技术概论[M],中国农业出版社,1999：26-41
王国莉,李希陶.苦瓜离体再生体系建立的研究[J].安徽农业科学,2008,36(11)：4408-4410
王纪方,金波等.蔬菜组织培养[M].上海科技出版社,1983
王开冻,魏跃.西瓜花药培养中不定芽的诱导与增殖[J].安徽农业科学,2008,36(16)：6826-6827
王立浩.辣椒花药培养中若干影响因素的研究[J].园艺学报,2004,31(2)：199-204
王立浩,张宝玺.辣椒花药培养研究进展[J].中国蔬菜,2001(3)：52-53
王伟光,高亦珂.花药培养的研究进展[J].内蒙古大学学报(自然科学版),2005,20(3)：280-284
王秀红,史向远,吴先军等.内源激素对水稻不同外植体培养力的影响[J].中国农业科学,2004,37(12)：1819-1823
王玉英,李春玲,蒋钟仁.辣椒和甜椒花药培养的新进展.园艺学报,1981,8(2)：41-45
王子霞,杨克锐,海热古力等.玉米花药培养的初步研究[J].新疆农业科学,2001,38(6)：346-347
肖关丽,杨清辉,李富生等.甘蔗愈伤组织分化绿苗内源激素变化规律研究[J].西南农业大学学报,2002,24(4)：337
谢淼,秦丽颖,潘俊松等.黄瓜花器形态发生、小孢子发育与花药培养[J].西北植物学报,2005,25(6)：1096-1100
宣朴,陈新,岳春芳等.苦瓜愈伤组织再生植株研究[J].西南农业大学报,2006,19(5),940-942
薛光荣,费开伟.西瓜花药离体培养获得花粉植株简报[J].中国果树,1982,(2)：51-52
颜昌敬.植物组织培养手册[M].上海,上海科学技术出版社,1990：108
杨学荣.水稻花培育种研究[M].北京,农业出版社,1983：61
杨一平,王述礼,曾士余等.杨树花粉发育途径及某些因素的影响[J].林业科学,1980,16(4)：257-263
叶兴国,王连铮.大豆花药愈伤组织的分化及其内源激素分析[J].作物学报,1997,23(5)：555-561
喻财铃.丝瓜小孢子囊与雄配子体发育的研究和花药离体培养初探[D].华南农业大学,2001
于伟常,沈利爽,文香玉.哈密瓜内源激素IAA的测定[C].中国科学院遗传研究所工作年报,1990,40-41
余叔文,汤章城.植物生理与分子生物学[M].北京,科学出版社,1999
袁美.花生花药培养的初步研究[J].花生学报,2004,33(4)：11-15
袁万良,付润民,雷保林等.西瓜花培试验初报[J].陕西农业科学,1995(1)：29-30
袁亦楠,朱德蔚,连勇等.番茄游离小孢子培养形成胚状体的初步研究[J].农业生物技术学报1997,(1)：85-88
张超美,张再君,张杏枝等.提高小麦花药培养花粉植株诱导率的途径[J].湖北农学院学报,1995,15(4)：304-305
张德双,曹鸣庆,秦智伟.绿菜花游离小孢子培养、胚胎发生和植株再生[J].华北农学报,1998,13(3)：102-106
张明方,蒋有条.番茄不同分化潜力愈伤组织中内源多胺比较及外源多胺对分化的影响[J].浙江农业大学学报,1997,23(6)：677-681
张鹏,傅爱根,王爱国.AgNO3在植物离体培养中的作用及可能的机制[J].植物生理学通讯,1997,33(5)：376-379
张廷红,杨随庄.硝酸银对小麦花药愈伤组织分化的影响[J].甘肃农业科技,2008(2)：28-29
朱海山.活性炭对玉米花药培养的影响[J].植物生理学通讯,1996,32(1)：16-18
Ana E.V., Ricardo J.O., Belen F. et.al.. Relationships between hormonal contents and the organogenic response in Pinus pinea cotyledons[J]. Plant Physiol Biochem,2001,39: 377-384
Ashok Kumar H.G., Murthy H.N.. Effect of sugars and amino acids on androgenesis of Cucumis sativus[J]. Plant Cell, Tissue and Organ Culture,2004,78:201-208
Ashok Kumar H.G., Murthy H.N., Paek K.Y.. Embryogenesis and plant regeneration from anther cultures of Cucumis sativus L[J]. Scientia Hoticulture,2003,98:213-222.
Barro F., Martin A.. Response of different genotypes of Brassica carinata to microspore culture[J]. Plant Breeding,1999,118 (1):79-81
Centeno M.L., Rodriguez A., Feito I.et.al.. Relationship between endogenous auxin and cytokinin levels and morphogenic responses in Actinidia deliciosa tissue cultures[J]. Plant Cell Rep,1996,16:58-62
Chang M.K., Conkerton E.J., Chapital D.C.. Chinese Melon (Momordica charantia L.) Seed:Composition and Potential Use[J]. Journal of the American Oil Chemists Society, 1996,73(2):263-265
Grover J.K. and Gupta S. R.. Hypoglycemic activity of seeds of Momordica charantia[J]. European Journal of Pharmacology,1990,183 (3):1026-1027
Grover J.K., Yadav S.P.. Pharmacological actions and potential uses of Momordica charantia: a review[J]. Journal of Ethnopharmacology,2004,93 (1):123-132
Hiroshi O., Koichi W., Shunpei U., In vitro morphogenetic response and distribution of endogenous plant hormones in hypocotyl segments of snapdragon (Antirrhinum majus L.)[J]. Plant Cell Rep,1991,10:501-504
Karsai L., Belo I., Hayes P. M.. Effect of induction medium pH and marlstone concentration on in vitro autogenesis of haploid winter critical and wheat[J]. Plant cell tissue and Org. Cult,1994,39:49-53
Lidia Dora Martinez & Ines Noher de Halac. Organogenesis of anther-derived calluses in long-term cultures of Oenothera hookeri de Vries[J]. Plant Cell Tiss Org Cult,1995, 42:91-96
Matsubara S., Hu K., Murakami K.. Embryoid and callus formation from pollen grains of eggplant and pepper by anther culture[J]. Journal of the Japanese Society for Horticultural Science,1992,61(1):69-77
Meskaoui E.I., Tremblay F.M.. Involvement of ethylene in the maturation of black spruce embryogenic cell lines with different maturation capacities[J]. J. Exp. Bot.,2001,52:761
Metwally E.L., Moustafa S.A., Shalaby T.A.. Haploid plantlets derived by anther culture of Cucurbitapepo[J]. Plant Cell, Tissue and Organ Culture,1998,52:171-176
Mityko J., Fari M., Altman A. et. al.. Problems and results of doubled haploid plant production in pepper (Capsicum annuum L.) via anther and microspore culture[J]. Act Horticultural,1997,447:281-287
Nadine Beloin, Messanvi Gbeassor, Koffi Akpagana et. al.. Ethnomedicinal uses of Momordica charantia (Cucurbitaceae) in Togo and relation to its phytochemistry and biological activity[J]. Journal of Ethnopharmacology,2005,96(1-2):49-55
Rimberia F.K., Sungagawa H., Urasaki N. et al..Embryo induction via anther culture in papaya and sex analysis of the derived plantlets[J]. Sci. hort.,2005,103:199-208
Sandal I., Kumar A., Bhattacharya A.. Gradual depletion of 2,4-D in the culture medium for indirect shoot regeneration from leaf explants of Camellia sinensis(L.) [J]. Plant Growth Regulation,2005,47:121
Thiruvengadam M., Varisai S., Mohamed et. al.. Development of an embryogenic suspension culture of bitter melo[J]. Scientia Horticulturae,2006,109:123-129
Thomas J.C., Vatteran F.R.. Cytokinin activity induced by thidiazuron[J]. Plant Physiol., 1986,81:681
Victor M.J., Bangerth F.. Endogenous hormone concentrations and embryogenic in wheat[J]. Plant Cell, Tissue and Organ Culture,2001,67:37
Walters, Walters D.. Notes on Economic Plants, Balsam Pear (Momordica charantia, Cucurbitaceae) [J]. Economic Botany,1988,42(2):286-289
Wang H.C., Chen J.T.,Wu S.P. et. al.. Plant regeneration through shoot formation from callus of Areca catechu [J]. Plant Cell, Tissue and Organ Culture,2003,75:95
Zhang Y.F., Zhou J.H., Wu T. et.al.. Shoot regeneration and the relationship between organogenic capacity and endogenous hormonal contents in pumpkin[J]. Plant Cell Tiss Org Cult,2008,93:323-331
曹家树,申书兴.园艺植物育种学[M].中国农业大学出版社,2001
曹鸣庆,李岩,刘凡.基因型和供体植株生长环境对大白菜游离小孢子胚胎生长的影响[J].华北农学报,1993,8(4)：1-6
曹毅,王月,刘薇.甘蓝型油菜花粉离体培养[J].西北植物学报,2010,30(2)：399-404
陈佳.苦瓜花药培养诱导愈伤组织及器官分化的研究[D].四川农业大学,2007
董艳荣.甜瓜花药再生体系的基础研究[D].南京农业大学,2002
何艳.苦瓜花药离体培养初探[D].四川农业大学,2008
缑艳霞,张明方.葫芦科花药培养再生体系和单倍体育种[J].北方园艺,2006(4)：63-66
栗根义,高睦枪,赵秀山.大白菜游离小孢子培养[J].园艺学报,1993,20(2)：167-170
孙爱新,赵飏,吕国华等.瓜尔豆小孢子发育时期与花器形态相关性研究[J].石河子大学学报(自然科学版),2004,22(4)：103-106
魏瑛,张俊莲,陈年来等.西瓜花药愈伤组织的诱导[J].甘肃农业大学学报,1998,33：127-130
王博.苦瓜花药培养愈伤组织诱导和分化研究[D].四川农业大学,2009
王玲仙,蔺忠龙等.水稻游离小孢子培养最新研究进展[J].生物技术,2009,9(6)：92-95
谢淼,秦丽颖,潘俊松等.黄瓜花器形态发生、小孢子发育与花药培养[J].西北植物学报,2005,25(6)：1096-1100
喻财铃.丝瓜小孢子囊与雄配子体发育的研究和花药离体培养初探[D].华南农业大学,2001
张德双,曹鸣庆,秦智伟.影响绿菜花游离小孢子培养的因素[J].华北农学报,1999,14(1)：68-72
朱惠琴,戴文新.烟草加倍单倍体群体的构建及其在遗传育种中的应用[J].青海大学学报(自然科学版),2003,21(6)：20-23
朱丽萍,陈火英,庄天明等.番茄不同花药长短及花粉发育时期与其愈伤组织诱导率的相关性[J].上海交通大学学报(农业科学版),2005,23(3)：239-243
Summers W.L., Jaramillo J., Bailey T.. Microspore developmental stage and anther length influence the induction of tomato anther callus[J]. Hort Science,1992,27(7):838-840
Tomasi P., Dierig D. A., Backhaus R. A. et.al.. Floral bud and mean petal length as morphological predictors of microspore cytological stage in Lesquerella[J]. Hort Sci.,1999, 34:1269-1270
Willcox M.C., Reed S.M., Burns J.A. et. al.. Effect of microspore stage and media on anther culture of peanut (Arachis hypogaea L.) [J]. Plant Cell, Tissue and Organ Culture,1991,24(1):25-28
曹禹,李鹏飞,甘玲玉等.冬性小黑麦花药培养的基因型和培养基效应的研究[J].新疆农业科学,2007(44)：97-100
陈红.枇杷花药愈伤组织诱导的研究[D].四川农业大学,2003
戴朝曦,冉毅东,王清等.细胞工程技术在马铃薯育种中应用的研究[J].遗传,1998,20(增刊)：39-42
董艳荣.甜瓜花药再生体系的基础研究[D].南京农业大学,2002
杜捷,王刚,幸亨泰等.兰州百合继代培养过程中的染色体变异[J].西北师范大学学报(自然科学版),2003,39(2)：61-65
黄晓梅,李桂英,梁艳等.组织培养中大蒜染色体倍性变异研究[J].中国农学通报,2005,21(1)：50-52
李娟,张丽,李焕秀等.西瓜花药培养技术研究[J].中国瓜菜,2008(4)：4-8
李俊强.枇杷花药胚胎发生和遗传转化研究[D].四川农业大学,2007
栾非时,陈典,崔喜波.大蒜茎尖愈伤组织的诱导、植株分化及变异的研究[J].北方园艺,1993,4(91)：23-25
马刘峰.甜瓜花药离体培养研究[D].石河子大学,2006
彭新红,周健松.芦笋花药培养胚状体诱导条件优化的初探[J].江西农业大学学报,2006,28(1)：39-43
魏瑛.低温预处理对西瓜花药愈伤组织诱导的影响[J].甘肃农业科技,1999,9：34-35
魏跃,龚义勤,邓波等.西瓜花药愈伤组织的诱导[J].湖北农业科学,2005(5)：93-95
王子霞,杨克锐,海热古力等.玉米花药培养的初步研究[J].新疆农业科学,2001,38(6)：346-347
宣朴,陈新,岳春芳等.苦瓜愈伤组织再生植株研究[J].西南农业大学报,2006,19(5),940-942
杨一平,王述礼,曾士余等.杨树花粉发育途径及某些因素的影响[J].林业科学,1980,16(4)：257-263
袁美.花生花药培养的初步研究[J].花生学报,2004,33(4)：11-15
袁万良,付润民,雷保林等.西瓜花培试验初报[J].陕西农业科学,1995(1)：29-30
张素芝,李纪蓉.植物激素对大蒜茎盘组织培养的影响[J].西南农业大学学报(自然 科学版),2006,28(5)：805-808
Ashok Kumar H.G., Murthy H.N., Paek K.Y.. Embryogenesis and plant regeneration from anther cultures of Cucumis sativus L[J]. Scientia Hoticulture,2003,98:213-222.
Ashok Kumar H.G., Murthy H.N.. Effect of sugars and amino acids on androgenesis of Cucumis sativus[J]. Plant Cell, Tissue and Organ Culture,2004,78:201-208
Duncan E.J. and Heberle-Bors E.. Effct of temperature shock on nuclear phenomena in microspores of Nicotiana rabacum and consequently on plantlet production[J]. Protoplasma,1976,90:173-177
Jacobsen E.. Polyploidization in leaf callus tissue amiprophos-methl and colchicines in potato. Plant Cell Tissue and Organ Culture,1981, (1):77-84
Keller W.A. and Armstrong K. C. Stimulation of embryogenesis in Brassica compestris anther cultures by elevated temperature treatments [J]. Theor. Appl. Genet,1979,55:65-67
Lazarte J.E., Sasser C.C., Asexual. Embryogenesis and plantlet development in anther culture of Cucumissativus L.[J]. Hortscience,1982,17:88
Martensson B., Widell S.. Pollen from cold-treated Nicotiana tabacum:Embryogenic capacity, peroxidase activity and partitioning in aqueous two-phase systems[J]. Plant Cell Tissue Organ Cult.,1993,35:141-149
Matsubara S., Hu K., Murakami K.. Embryoid and callus formation from pollen grains of eggplant and pepper by anther culture[J]. Journal of the Japanese Society for Horticultural Science,1992,61(1):69-77
Metwally E.L., Moustafa S.A., Shalaby T.A.. Haploid plantlets derived by anther culture of Cucurbitapepo[J]. Plant Cell, Tissue and Organ Culture,1998,52:171-176
Mityko J., Fari M., Altman A. et. al.. Problems and results of doubled haploid plant production in pepper (Capsicum annuum L.) via anther and microspore culture[J]. Act Horticultural,1997,447:281-287
Sandal I., Kumar A., Bhartacharya A.. Gradual depletion of 2,4-D in the culture medium for indirect shoot regeneration from leaf explants of Camellia sinensis(L.) [J]. Plant Growth Regulation,2005,47:121
Sunderland N.. Strategies in the improvement of yield in anther culture[J]. In proceedings of symposium on plant tissue culture. Peking Science Press,1978,65-86
Thiruvengadam M., Varisai S., Mohamed et.al.. Development of an embryogenic suspension culture of bitter melo[J]. Scientia Horticulturae,2006,109:123-129
Wenzel G., Hoffmann F., Thomas E.. Increased induction and chromosome doubling of androgenetic haploid rye[J]. Theor. Appl. Genet.,1977,51:81-86
陈远平,杨文钰.卵叶韭休眠芽中GA3、IAA、ABA和ZT的高效液相色谱法测定[J].四川农业大学学报,2005,23(4)：498-500
盛艳萍,杨建平,张松等.大葱成熟胚离体再生过程中内源激素的变化[J].园艺学报.2005,32(3)：318-320
付凤玲,冯质雷,渠柏艳等.玉米未成熟胚胚性愈伤组织诱导率与内源激素含量的关系[J].核农学报,2006,20(1)：10
郭子彪,盖均镒.内源激素IAA、ABA对大豆萌发子叶胚性愈伤组织诱导及其分化的调控[J].大豆科学,1997,16(3)：194-198
李雪梅,刘熔山.小麦幼穗胚性愈伤组织诱导及分化过程中内源激素的作用[J].植物生理学通讯,1994,30(4)：255-260
罗琼,胡延玉,周开达.内源激素对水稻成熟胚培养力的影响[J].中国水稻科学,1998,12(4)：238-240
宋莉英,高峰.苦瓜离体培养过程中内源激素含量的变化[J].植物学通报,2006,23(2)：192-196
汤国庆,李文安.在离体条件下拐芹形成愈伤组织和非胚性愈伤组织过程中几种内源激素的变化[J].实验生物学报,1995,28(2)：203
唐玉林,陈婉芳,周燮.烟草叶块分化根和芽过程中内源激素水平的变化[J].南京农业大学学报,1996,19(2)：12-16
陶静,詹亚光,由香玲等.白桦组培再生系统的研究[J].东北林业大学学报,1998,26(6)：6-9
王秀红,史向远,吴先军等.内源激素对水稻不同外植体培养力的影响[J].中国农业科学,2004,37(12)：1819-1823
肖关丽,杨清辉,李富生等.甘蔗愈伤组织分化绿苗内源激素变化规律研究[J].西南农业大学学报,2002,24(4)：337
叶兴国,王连铮.大豆花药愈伤组织的分化及其内源激素分析[J].作物学报,1997,23(5)：555-561
赵晓菊,唐中华,郭晓瑞等.固相萃取富集-高效液相色谱法测定长春花中的3种内源激素[J].色谱,2006,24(5)：534
Ana E.V., Ricardo J.O., Belen F. et.al. Relationships between hormonal contents and the organogenic response in Pinus pinea cotyledons[J]. Plant Physiol Biochem,2001,39: 377-384
Branca C., Bucci G., Domiano P. et. al..Auxin structure and activity on tomato morphogenesis in vitro and pea stem elongation[J]. Plant Cell Tiss Org Cult,1991, 24:105-114
Centeno M.L., Rodriguez A., Feito I.et.al.. Relationship between endogenous auxin and cytokinin levels and morphogenic responses in Actinidia deliciosa tissue cultures[J]. Plant Cell Rep,1996,16:58-62
Hiroshi O., Koichi W., Shunpei U., In vitro morphogenetic response and distribution of endogenous plant hormones in hypocotyl segments of snapdragon(Antirrhinum majus L.)[J]. Plant Cell Rep,1991,10:501-504
Kopertekh L.G.& Butenko R.G.. Naturally occurring phytohormones in wheat explants as related to wheat morphogenesis in vitro[J]. Rus. J. Plant Physiol,1995,42:488^4-91
Lidia Dora Martinez & Ines Noher de Halac. Organogenesis of anther-derived calluses in long-term cultures of Oenothera hookeri de Vries[J]. Plant Cell Tiss Org Cult,1995, 42:91-96
Sarul P., Vlahova M., Ivanova A.et.al.. Direct shoot formation in spontaneously occurring root pseudonodules of alfalfa(Medicago sativa L.) [J]. In Vitro Cell Dev. Biol.,1995,31: 21-25
Victor M.J.. Involvement of plant hormones and plant growth regulators on in vitro somatic embryogenesis[J]. Plant Growth Regulation,2005,47:91-110
Victor M.J., Bangerth F.. Endogenous hormone concentrations and embryogenic in wheat[J]. Plant Cell, Tissue and Organ Culture,2001,67:37
Yoshimatsu K., Shimomura K.. Plant regeneration on cultured roots segments of Cephalis ipecacuanha A. Richard[J]. Plant Cell Rep,1994,14:98-101
Zaffari G.R., Kerbauy G.B., Kraus J.E.et.al.. Hormonal and histological studies related to in vitro banana bud formation[J]. Plant Cell Tiss Org Cult,2000,63:187-192
Zhang Y.F., Zhou J.H., Wu T. et.al.. Shoot regeneration and the relationship between organogenic capacity and endogenous hormonal contents in pumpkin[J]. Plant Cell Tiss Org Cult,2008,93:323-331
曹利仙,赵鹉,唐宇力等.硝酸银对黄瓜离体子叶培养芽再生的促进效应[J].甘肃农业大学学报,2001,36(2)：168-171
陈佳.苦瓜花药培养诱导愈伤组织及器官分化的研究[D].四川农业大学,2007
陈晓,詹玉丝,徐小利等.花药培养建立辣椒DH纯系的初步研究[J].河南农业科学,2003(3)：52-55
陈振光.园艺植物离体培养学[M].中国农业出版社,1995,23-25
盛艳萍,杨建平,张松等.大葱成熟胚离体再生过程中内源激素的变化[J].园艺学报.2005,32(3)：318-320
代色平,包满珠.矮牵牛花药培养及植株再生研究[J].亚热带植物科学,2003,32(2)：55-57
邓向东,耿玉轩,路子显等.外植体和培养因子对哈密瓜不定芽诱导的影响[J].园艺学报,1996,23(1)：57-61
董艳荣.甜瓜花药再生体系的基础研究[D].南京农业大学,2002
冯玉生,王则僖.紫云英植物诱导初报[J].科学通报,1981,3：30-34
付凤玲,冯质雷,渠柏艳等.玉米未成熟胚胚性愈伤组织诱导率与内源激素含量的关系[J].核农学报,2006,20(1)：10
耿晨光,何刚.瓜类花药培养中激素和碳源的应用研究进展[J].长江蔬菜,2008,7b：5-9
郭子彪,盖均镒.内源激素IAA、ABA对大豆萌发子叶胚性愈伤组织诱导及其分化的调控[J].大豆科学,1997,16(3)：194-198
韩丽华.厚皮甜瓜未受精胚珠离体培养技术[D].河北农业大学,2004
何晓明,谢大森,陈清华等.我国瓜类细胞工程研究进展[J].深圳职业技术学院学报,2002(3)：18-19
何艳.苦瓜花药离体培养初探[D].四川农业大学,2008
何艳,李焕秀,郑阳霞等.苦瓜花药培养诱导愈伤组织的研究[J].北方园艺,2008(12)：164-16
缑艳霞.西瓜花药离体培养技术研究[D].浙江大学,2006
黄荣韶,覃伟.无籽西瓜离体培养中玻璃化现象的研究[J].广西农业大学学报,1998,17(3)：266-270
黄泰康,丁志遵,赵守训等.现代本草纲目(上卷)[M].中国医药科技出版社,2000
季彪俊,江树业,陈启锋等.活性炭在水稻花药培养中的作用[J].福建农业大学学报,1998,27(1)：16-19
李焕秀,Murch S.J., Sacena P.K.. TDZ诱导的黄芩外植体再生研究[J].四川农业大学学报,2000,18(4)：325
李娟.西瓜花药和小孢子培养技术研究[D].四川农业大学,2008
李娟,张丽,李焕秀等.西瓜花药培养技术研究[J].中国瓜菜,2008(4)：4-8
李俊明.植物组织培养教程[M].北京,北京农业大学出版社,1992
李文泽,景健康,姚根怀等.基因型和甘露醇预处理对大麦花粉植株高频率再生的影响[J].科学通报,1992,10：935-938
李文泽,宋子红,景健康等.甘露醇预处理对大麦雄核发育的影响[J].植物学报,1995,37(7)：552-557
李锡香,沈镝,王海平.我国蔬菜种质资源研究进展与发展策略[J].中国蔬菜,2006(增刊)：3-9
李雪梅,刘熔山.小麦幼穗胚性愈伤组织诱导及分化过程中内源激素的作用[J].植物生理学通讯,1994,30(4)：255-260
李宗霆,周燮.植物激素及其免疫检测技术[M].南京,江苏科学技术出版社,1996
刘涤,迟静芬,刘桂芸.烟草愈伤组织器官发生过程中外源激素的作用[J].植物生理与分子生物学报,1986,12(1)：104
刘贵仁,严仁玲,张磊等.石刁柏花药培养与染色体倍性的观察[J].吉林农业大学学报,1992,12(1)：5-8
刘京贞.天竺葵的应用及组织培养[J].北方园艺,2002(2)：54-56
罗言云.麻疯树花药培养及两个核糖体失活蛋白基因的表达研究[D].四川大学,2006
马刘峰.甜瓜花药离体培养研究[D].石河子大学,2006
马刘峰等.甜瓜花药愈伤组织分化与激素处理间的关系[J].中国瓜菜,2006(2)：4-6
潘瑞炽.植物组织培养[M].广东高等教育出版社,2001
彭爱芝,李劲.苦瓜中氨基酸和无机元素的含量分析[J].湖南医科大学学报,1996,21(4)：305-307
彭新红,周健松.芦笋花药培养胚状体诱导条件优化的初探[J].江西农业大学学报,2006,28(1)：39-43
戚春章,胡是麟,漆小泉.中国蔬菜种质资源的种类及分布[J].作物品种资源,1997,(1)：1-5
宋莉英,高峰.苦瓜离体培养过程中内源激素含量的变化[J].植物学通报,2006,(2)：66-70
苏华,金宝燕,任华中.黄瓜花药培养中若干影响因素的研究[J].沈阳农业大学学报,2006,37(3)：470-472
孙兰英,卢淑雯.黄瓜组织培养研究进展[J].黑龙江农业科学,2004,(4)：25-27
汤国庆,李文安.在离体条件下拐芹形成愈伤组织和非胚性愈伤组织过程中几种内源激素的变化[J].实验生物学报,1995,28(2)：203
唐玉林,陈婉芳,周燮.烟草叶块分化根和芽过程中内源激素水平的变化[J].南京农业大学学报,1996,19(2)：12-16
陶正南.甜瓜花药培养诱导成植株[J].植物生理学通讯,1987,(5)：43
田文忠.提高籼稻愈伤组织再生频率的研究[J].遗传学报,1994,21(3)：215
魏瑛.低温预处理对西瓜花药愈伤组织诱导的影响[J].甘肃农业科技,1999,9：34-35
魏瑛,张俊莲,陈年来等.西瓜花药愈伤组织的诱导[J].甘肃农业大学学报,1998,33：127-130
王得元,何晓明,王鸣.蔬菜生物技术概论[M],中国农业出版社,1999：26-41
王国莉,李希陶.苦瓜离体再生体系建立的研究[J].安徽农业科学,2008,36(11)：4408-4410
王纪方,金波等.蔬菜组织培养[M].上海科技出版社,1983
王开冻,魏跃.西瓜花药培养中不定芽的诱导与增殖[J].安徽农业科学,2008,36(16)：6826-6827
王立浩.辣椒花药培养中若干影响因素的研究[J].园艺学报,2004,31(2)：199-204
王立浩,张宝玺.辣椒花药培养研究进展[J].中国蔬菜,2001(3)：52-53
王伟光,高亦珂.花药培养的研究进展[J].内蒙古大学学报(自然科学版),2005,20(3)：280-284
王秀红,史向远,吴先军等.内源激素对水稻不同外植体培养力的影响[J].中国农业科学,2004,37(12)：1819-1823
王玉英,李春玲,蒋钟仁.辣椒和甜椒花药培养的新进展.园艺学报,1981,8(2)：41-45
王子霞,杨克锐,海热古力等.玉米花药培养的初步研究[J].新疆农业科学,2001,38(6)：346-347
肖关丽,杨清辉,李富生等.甘蔗愈伤组织分化绿苗内源激素变化规律研究[J].西南农业大学学报,2002,24(4)：337
谢淼,秦丽颖,潘俊松等.黄瓜花器形态发生、小孢子发育与花药培养[J].西北植物学报,2005,25(6)：1096-1100
宣朴,陈新,岳春芳等.苦瓜愈伤组织再生植株研究[J].西南农业大学报,2006,19(5),940-942
薛光荣,费开伟.西瓜花药离体培养获得花粉植株简报[J].中国果树,1982,(2)：51-52
颜昌敬.植物组织培养手册[M].上海,上海科学技术出版社,1990：108
杨学荣.水稻花培育种研究[M].北京,农业出版社,1983：61
杨一平,王述礼,曾士余等.杨树花粉发育途径及某些因素的影响[J].林业科学,1980,16(4)：257-263
叶兴国,王连铮.大豆花药愈伤组织的分化及其内源激素分析[J].作物学报,1997,23(5)：555-561
喻财铃.丝瓜小孢子囊与雄配子体发育的研究和花药离体培养初探[D].华南农业大学,2001
于伟常,沈利爽,文香玉.哈密瓜内源激素IAA的测定[C].中国科学院遗传研究所工作年报,1990,40-41
余叔文,汤章城.植物生理与分子生物学[M].北京,科学出版社,1999
袁美.花生花药培养的初步研究[J].花生学报,2004,33(4)：11-15
袁万良,付润民,雷保林等.西瓜花培试验初报[J].陕西农业科学,1995(1)：29-30
袁亦楠,朱德蔚,连勇等.番茄游离小孢子培养形成胚状体的初步研究[J].农业生物技术学报1997,(1)：85-88
张超美,张再君,张杏枝等.提高小麦花药培养花粉植株诱导率的途径[J].湖北农学院学报,1995,15(4)：304-305
张德双,曹鸣庆,秦智伟.绿菜花游离小孢子培养、胚胎发生和植株再生[J].华北农学报,1998,13(3)：102-106
张明方,蒋有条.番茄不同分化潜力愈伤组织中内源多胺比较及外源多胺对分化的影响[J].浙江农业大学学报,1997,23(6)：677-681
张鹏,傅爱根,王爱国.AgNO3在植物离体培养中的作用及可能的机制[J].植物生理学通讯,1997,33(5)：376-379
张廷红,杨随庄.硝酸银对小麦花药愈伤组织分化的影响[J].甘肃农业科技,2008(2)：28-29
朱海山.活性炭对玉米花药培养的影响[J].植物生理学通讯,1996,32(1)：16-18
Ana E.V., Ricardo J.O., Belen F. et.al.. Relationships between hormonal contents and the organogenic response in Pinus pinea cotyledons[J]. Plant Physiol Biochem,2001,39: 377-384
Ashok Kumar H.G., Murthy H.N.. Effect of sugars and amino acids on androgenesis of Cucumis sativus[J]. Plant Cell, Tissue and Organ Culture,2004,78:201-208
Ashok Kumar H.G., Murthy H.N., Paek K.Y.. Embryogenesis and plant regeneration from anther cultures of Cucumis sativus L[J]. Scientia Hoticulture,2003,98:213-222.
Barro F., Martin A.. Response of different genotypes of Brassica carinata to microspore culture[J]. Plant Breeding,1999,118 (1):79-81
Centeno M.L., Rodriguez A., Feito I.et.al.. Relationship between endogenous auxin and cytokinin levels and morphogenic responses in Actinidia deliciosa tissue cultures[J]. Plant Cell Rep,1996,16:58-62
Chang M.K., Conkerton E.J., Chapital D.C.. Chinese Melon (Momordica charantia L.) Seed:Composition and Potential Use[J]. Journal of the American Oil Chemists Society, 1996,73(2):263-265
Grover J.K. and Gupta S. R.. Hypoglycemic activity of seeds of Momordica charantia[J]. European Journal of Pharmacology,1990,183 (3):1026-1027
Grover J.K., Yadav S.P.. Pharmacological actions and potential uses of Momordica charantia: a review[J]. Journal of Ethnopharmacology,2004,93 (1):123-132
Hiroshi O., Koichi W., Shunpei U., In vitro morphogenetic response and distribution of endogenous plant hormones in hypocotyl segments of snapdragon (Antirrhinum majus L.)[J]. Plant Cell Rep,1991,10:501-504
Karsai L., Belo I., Hayes P. M.. Effect of induction medium pH and marlstone concentration on in vitro autogenesis of haploid winter critical and wheat[J]. Plant cell tissue and Org. Cult,1994,39:49-53
Lidia Dora Martinez & Ines Noher de Halac. Organogenesis of anther-derived calluses in long-term cultures of Oenothera hookeri de Vries[J]. Plant Cell Tiss Org Cult,1995, 42:91-96
Matsubara S., Hu K., Murakami K.. Embryoid and callus formation from pollen grains of eggplant and pepper by anther culture[J]. Journal of the Japanese Society for Horticultural Science,1992,61(1):69-77
Meskaoui E.I., Tremblay F.M.. Involvement of ethylene in the maturation of black spruce embryogenic cell lines with different maturation capacities[J]. J. Exp. Bot.,2001,52:761
Metwally E.L., Moustafa S.A., Shalaby T.A.. Haploid plantlets derived by anther culture of Cucurbitapepo[J]. Plant Cell, Tissue and Organ Culture,1998,52:171-176
Mityko J., Fari M., Altman A. et. al.. Problems and results of doubled haploid plant production in pepper (Capsicum annuum L.) via anther and microspore culture[J]. Act Horticultural,1997,447:281-287
Nadine Beloin, Messanvi Gbeassor, Koffi Akpagana et. al.. Ethnomedicinal uses of Momordica charantia (Cucurbitaceae) in Togo and relation to its phytochemistry and biological activity[J]. Journal of Ethnopharmacology,2005,96(1-2):49-55
Rimberia F.K., Sungagawa H., Urasaki N. et al..Embryo induction via anther culture in papaya and sex analysis of the derived plantlets[J]. Sci. hort.,2005,103:199-208
Sandal I., Kumar A., Bhattacharya A.. Gradual depletion of 2,4-D in the culture medium for indirect shoot regeneration from leaf explants of Camellia sinensis(L.) [J]. Plant Growth Regulation,2005,47:121
Thiruvengadam M., Varisai S., Mohamed et. al.. Development of an embryogenic suspension culture of bitter melo[J]. Scientia Horticulturae,2006,109:123-129
Thomas J.C., Vatteran F.R.. Cytokinin activity induced by thidiazuron[J]. Plant Physiol., 1986,81:681
Victor M.J., Bangerth F.. Endogenous hormone concentrations and embryogenic in wheat[J]. Plant Cell, Tissue and Organ Culture,2001,67:37
Walters, Walters D.. Notes on Economic Plants, Balsam Pear (Momordica charantia, Cucurbitaceae) [J]. Economic Botany,1988,42(2):286-289
Wang H.C., Chen J.T.,Wu S.P. et. al.. Plant regeneration through shoot formation from callus of Areca catechu [J]. Plant Cell, Tissue and Organ Culture,2003,75:95
Zhang Y.F., Zhou J.H., Wu T. et.al.. Shoot regeneration and the relationship between organogenic capacity and endogenous hormonal contents in pumpkin[J]. Plant Cell Tiss Org Cult,2008,93:323-331
曹家树,申书兴.园艺植物育种学[M].中国农业大学出版社,2001
曹鸣庆,李岩,刘凡.基因型和供体植株生长环境对大白菜游离小孢子胚胎生长的影响[J].华北农学报,1993,8(4)：1-6
曹毅,王月,刘薇.甘蓝型油菜花粉离体培养[J].西北植物学报,2010,30(2)：399-404
陈佳.苦瓜花药培养诱导愈伤组织及器官分化的研究[D].四川农业大学,2007
董艳荣.甜瓜花药再生体系的基础研究[D].南京农业大学,2002
何艳.苦瓜花药离体培养初探[D].四川农业大学,2008
缑艳霞,张明方.葫芦科花药培养再生体系和单倍体育种[J].北方园艺,2006(4)：63-66
栗根义,高睦枪,赵秀山.大白菜游离小孢子培养[J].园艺学报,1993,20(2)：167-170
孙爱新,赵飏,吕国华等.瓜尔豆小孢子发育时期与花器形态相关性研究[J].石河子大学学报(自然科学版),2004,22(4)：103-106
魏瑛,张俊莲,陈年来等.西瓜花药愈伤组织的诱导[J].甘肃农业大学学报,1998,33：127-130
王博.苦瓜花药培养愈伤组织诱导和分化研究[D].四川农业大学,2009
王玲仙,蔺忠龙等.水稻游离小孢子培养最新研究进展[J].生物技术,2009,9(6)：92-95
谢淼,秦丽颖,潘俊松等.黄瓜花器形态发生、小孢子发育与花药培养[J].西北植物学报,2005,25(6)：1096-1100
喻财铃.丝瓜小孢子囊与雄配子体发育的研究和花药离体培养初探[D].华南农业大学,2001
张德双,曹鸣庆,秦智伟.影响绿菜花游离小孢子培养的因素[J].华北农学报,1999,14(1)：68-72
朱惠琴,戴文新.烟草加倍单倍体群体的构建及其在遗传育种中的应用[J].青海大学学报(自然科学版),2003,21(6)：20-23
朱丽萍,陈火英,庄天明等.番茄不同花药长短及花粉发育时期与其愈伤组织诱导率的相关性[J].上海交通大学学报(农业科学版),2005,23(3)：239-243
Summers W.L., Jaramillo J., Bailey T.. Microspore developmental stage and anther length influence the induction of tomato anther callus[J]. Hort Science,1992,27(7):838-840
Tomasi P., Dierig D. A., Backhaus R. A. et.al.. Floral bud and mean petal length as morphological predictors of microspore cytological stage in Lesquerella[J]. Hort Sci.,1999, 34:1269-1270
Willcox M.C., Reed S.M., Burns J.A. et. al.. Effect of microspore stage and media on anther culture of peanut (Arachis hypogaea L.) [J]. Plant Cell, Tissue and Organ Culture,1991,24(1):25-28
曹禹,李鹏飞,甘玲玉等.冬性小黑麦花药培养的基因型和培养基效应的研究[J].新疆农业科学,2007(44)：97-100
陈红.枇杷花药愈伤组织诱导的研究[D].四川农业大学,2003
戴朝曦,冉毅东,王清等.细胞工程技术在马铃薯育种中应用的研究[J].遗传,1998,20(增刊)：39-42
董艳荣.甜瓜花药再生体系的基础研究[D].南京农业大学,2002
杜捷,王刚,幸亨泰等.兰州百合继代培养过程中的染色体变异[J].西北师范大学学报(自然科学版),2003,39(2)：61-65
黄晓梅,李桂英,梁艳等.组织培养中大蒜染色体倍性变异研究[J].中国农学通报,2005,21(1)：50-52
李娟,张丽,李焕秀等.西瓜花药培养技术研究[J].中国瓜菜,2008(4)：4-8
李俊强.枇杷花药胚胎发生和遗传转化研究[D].四川农业大学,2007
栾非时,陈典,崔喜波.大蒜茎尖愈伤组织的诱导、植株分化及变异的研究[J].北方园艺,1993,4(91)：23-25
马刘峰.甜瓜花药离体培养研究[D].石河子大学,2006
彭新红,周健松.芦笋花药培养胚状体诱导条件优化的初探[J].江西农业大学学报,2006,28(1)：39-43
魏瑛.低温预处理对西瓜花药愈伤组织诱导的影响[J].甘肃农业科技,1999,9：34-35
魏跃,龚义勤,邓波等.西瓜花药愈伤组织的诱导[J].湖北农业科学,2005(5)：93-95
王子霞,杨克锐,海热古力等.玉米花药培养的初步研究[J].新疆农业科学,2001,38(6)：346-347
宣朴,陈新,岳春芳等.苦瓜愈伤组织再生植株研究[J].西南农业大学报,2006,19(5),940-942
杨一平,王述礼,曾士余等.杨树花粉发育途径及某些因素的影响[J].林业科学,1980,16(4)：257-263
袁美.花生花药培养的初步研究[J].花生学报,2004,33(4)：11-15
袁万良,付润民,雷保林等.西瓜花培试验初报[J].陕西农业科学,1995(1)：29-30
张素芝,李纪蓉.植物激素对大蒜茎盘组织培养的影响[J].西南农业大学学报(自然 科学版),2006,28(5)：805-808
Ashok Kumar H.G., Murthy H.N., Paek K.Y.. Embryogenesis and plant regeneration from anther cultures of Cucumis sativus L[J]. Scientia Hoticulture,2003,98:213-222.
Ashok Kumar H.G., Murthy H.N.. Effect of sugars and amino acids on androgenesis of Cucumis sativus[J]. Plant Cell, Tissue and Organ Culture,2004,78:201-208
Duncan E.J. and Heberle-Bors E.. Effct of temperature shock on nuclear phenomena in microspores of Nicotiana rabacum and consequently on plantlet production[J]. Protoplasma,1976,90:173-177
Jacobsen E.. Polyploidization in leaf callus tissue amiprophos-methl and colchicines in potato. Plant Cell Tissue and Organ Culture,1981, (1):77-84
Keller W.A. and Armstrong K. C. Stimulation of embryogenesis in Brassica compestris anther cultures by elevated temperature treatments [J]. Theor. Appl. Genet,1979,55:65-67
Lazarte J.E., Sasser C.C., Asexual. Embryogenesis and plantlet development in anther culture of Cucumissativus L.[J]. Hortscience,1982,17:88
Martensson B., Widell S.. Pollen from cold-treated Nicotiana tabacum:Embryogenic capacity, peroxidase activity and partitioning in aqueous two-phase systems[J]. Plant Cell Tissue Organ Cult.,1993,35:141-149
Matsubara S., Hu K., Murakami K.. Embryoid and callus formation from pollen grains of eggplant and pepper by anther culture[J]. Journal of the Japanese Society for Horticultural Science,1992,61(1):69-77
Metwally E.L., Moustafa S.A., Shalaby T.A.. Haploid plantlets derived by anther culture of Cucurbitapepo[J]. Plant Cell, Tissue and Organ Culture,1998,52:171-176
Mityko J., Fari M., Altman A. et. al.. Problems and results of doubled haploid plant production in pepper (Capsicum annuum L.) via anther and microspore culture[J]. Act Horticultural,1997,447:281-287
Sandal I., Kumar A., Bhartacharya A.. Gradual depletion of 2,4-D in the culture medium for indirect shoot regeneration from leaf explants of Camellia sinensis(L.) [J]. Plant Growth Regulation,2005,47:121
Sunderland N.. Strategies in the improvement of yield in anther culture[J]. In proceedings of symposium on plant tissue culture. Peking Science Press,1978,65-86
Thiruvengadam M., Varisai S., Mohamed et.al.. Development of an embryogenic suspension culture of bitter melo[J]. Scientia Horticulturae,2006,109:123-129
Wenzel G., Hoffmann F., Thomas E.. Increased induction and chromosome doubling of androgenetic haploid rye[J]. Theor. Appl. Genet.,1977,51:81-86
陈远平,杨文钰.卵叶韭休眠芽中GA3、IAA、ABA和ZT的高效液相色谱法测定[J].四川农业大学学报,2005,23(4)：498-500
盛艳萍,杨建平,张松等.大葱成熟胚离体再生过程中内源激素的变化[J].园艺学报.2005,32(3)：318-320
付凤玲,冯质雷,渠柏艳等.玉米未成熟胚胚性愈伤组织诱导率与内源激素含量的关系[J].核农学报,2006,20(1)：10
郭子彪,盖均镒.内源激素IAA、ABA对大豆萌发子叶胚性愈伤组织诱导及其分化的调控[J].大豆科学,1997,16(3)：194-198
李雪梅,刘熔山.小麦幼穗胚性愈伤组织诱导及分化过程中内源激素的作用[J].植物生理学通讯,1994,30(4)：255-260
罗琼,胡延玉,周开达.内源激素对水稻成熟胚培养力的影响[J].中国水稻科学,1998,12(4)：238-240
宋莉英,高峰.苦瓜离体培养过程中内源激素含量的变化[J].植物学通报,2006,23(2)：192-196
汤国庆,李文安.在离体条件下拐芹形成愈伤组织和非胚性愈伤组织过程中几种内源激素的变化[J].实验生物学报,1995,28(2)：203
唐玉林,陈婉芳,周燮.烟草叶块分化根和芽过程中内源激素水平的变化[J].南京农业大学学报,1996,19(2)：12-16
陶静,詹亚光,由香玲等.白桦组培再生系统的研究[J].东北林业大学学报,1998,26(6)：6-9
王秀红,史向远,吴先军等.内源激素对水稻不同外植体培养力的影响[J].中国农业科学,2004,37(12)：1819-1823
肖关丽,杨清辉,李富生等.甘蔗愈伤组织分化绿苗内源激素变化规律研究[J].西南农业大学学报,2002,24(4)：337
叶兴国,王连铮.大豆花药愈伤组织的分化及其内源激素分析[J].作物学报,1997,23(5)：555-561
赵晓菊,唐中华,郭晓瑞等.固相萃取富集-高效液相色谱法测定长春花中的3种内源激素[J].色谱,2006,24(5)：534
Ana E.V., Ricardo J.O., Belen F. et.al. Relationships between hormonal contents and the organogenic response in Pinus pinea cotyledons[J]. Plant Physiol Biochem,2001,39: 377-384
Branca C., Bucci G., Domiano P. et. al..Auxin structure and activity on tomato morphogenesis in vitro and pea stem elongation[J]. Plant Cell Tiss Org Cult,1991, 24:105-114
Centeno M.L., Rodriguez A., Feito I.et.al.. Relationship between endogenous auxin and cytokinin levels and morphogenic responses in Actinidia deliciosa tissue cultures[J]. Plant Cell Rep,1996,16:58-62
Hiroshi O., Koichi W., Shunpei U., In vitro morphogenetic response and distribution of endogenous plant hormones in hypocotyl segments of snapdragon(Antirrhinum majus L.)[J]. Plant Cell Rep,1991,10:501-504
Kopertekh L.G.& Butenko R.G.. Naturally occurring phytohormones in wheat explants as related to wheat morphogenesis in vitro[J]. Rus. J. Plant Physiol,1995,42:488^4-91
Lidia Dora Martinez & Ines Noher de Halac. Organogenesis of anther-derived calluses in long-term cultures of Oenothera hookeri de Vries[J]. Plant Cell Tiss Org Cult,1995, 42:91-96
Sarul P., Vlahova M., Ivanova A.et.al.. Direct shoot formation in spontaneously occurring root pseudonodules of alfalfa(Medicago sativa L.) [J]. In Vitro Cell Dev. Biol.,1995,31: 21-25
Victor M.J.. Involvement of plant hormones and plant growth regulators on in vitro somatic embryogenesis[J]. Plant Growth Regulation,2005,47:91-110
Victor M.J., Bangerth F.. Endogenous hormone concentrations and embryogenic in wheat[J]. Plant Cell, Tissue and Organ Culture,2001,67:37
Yoshimatsu K., Shimomura K.. Plant regeneration on cultured roots segments of Cephalis ipecacuanha A. Richard[J]. Plant Cell Rep,1994,14:98-101
Zaffari G.R., Kerbauy G.B., Kraus J.E.et.al.. Hormonal and histological studies related to in vitro banana bud formation[J]. Plant Cell Tiss Org Cult,2000,63:187-192
Zhang Y.F., Zhou J.H., Wu T. et.al.. Shoot regeneration and the relationship between organogenic capacity and endogenous hormonal contents in pumpkin[J]. Plant Cell Tiss Org Cult,2008,93:323-331