四季花龙眼花芽分化期营养动态变化的研究
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摘要
四季花龙眼具有周年开花结果的习性、坐果率高、带蜂蜜风味的肉质果实等优点。在很多方面优于常规品种,其树姿开张、树体矮化,适于高密度栽培种植,所以研究四季花龙眼的花芽分化的过程和影响因素,是合理调控花芽分化进程和提高栽培效益的基础。
本试验以5-6a的四季花龙眼为试材,研究了冬梢花芽分化进程中碳素营养和相关生理活性的变化动态。采用石蜡切片法显微观察其花芽的形态分化,统计各分化时期的花芽分化率,测定其芽体和叶片的可溶性总糖、蔗糖、果糖、还原糖、淀粉和蛋白质含量及淀粉酶和过氧化物酶活性,分析了可溶性蛋白PAGE图谱、淀粉酶同工酶谱和过氧化物酶同工酶谱。从对比的角度探讨了不同器官营养物质变化与花芽分化的关系。结果表明：
1、四季花龙眼花芽形态分化过程可划分为未分化时期、花序原基分化期、花序主轴分化期、多级侧花序快速分化期和花器官分化期。在整个花芽分化过程中,顶芽多处于休眠或花芽和叶芽的中间状态,不能作为花芽分化起始的标志,而基部腋芽从露红点开始,较顶芽率先进入花芽分化期,且各分化时期有明显的阶段性,所以花芽分化应以腋芽形态分化为准。
2、在花芽分化期,叶片和芽体的各种可溶性糖含量的变化均呈现“低-高-低”的变化趋势,但芽体含量的变化幅度较大,其含量也高于叶片。叶片内淀粉含量变化呈显著的降低趋势而芽体内淀粉含量则呈上升趋势。与叶片相比较,芽体中的可溶性总糖、还原糖和蔗糖的含量变化与花芽分化的相关度更高,而芽体和叶片的淀粉含量与花芽分化率无显著相关性,说明芽体内较高的可溶性糖有利于花芽分化的进行,而淀粉积累水平并非成花的决定性因素。
3、成花过程中,成熟叶片的蛋白质含量,α-淀粉酶(α-AMY)活性和过氧化物酶(POD)活性均高于嫩叶含量,并在成花期有明显的变化规律,说明这些因素在成花过程中起到一定的积极意义。可溶性蛋白的SDS-PAGE凝胶电泳分析呈现有固定蛋白条带220KD,80KD和55KD,在花序主轴分化期和花器官快速分化期出现较为清晰的特异蛋白条带30KD和25KD。α-AMY同工酶酶谱呈现出逐渐增亮的条带。POD同工酶酶谱在花芽分化初期和花序原基分化期出现了新的条带,可作为分化的起始标志。它们的酶表达量的差异引起了酶活性的变化,说明SDS-PAGE和PAGE凝胶电泳呈现的条带变化与四季花龙眼花芽分化进程有密切的关系。
Sijihua longan has many advantages which are the habit of anniversary Blossoming, high fruit set, fleshy fruits with flavor of honey and so on. Its many aspects are superior to regular varieties, which include tree vigor Opening, tree stunting and the plant with high density. So to research the process and impact physiologic factors of flower bud differentiation is foundation of reasonable regulate flower bud differentiation process and improve the cultivation benefits.
Five to six-year-old sijihua longan were used as the experimental materials to research the change of nutrition dynamic during flower bud differentiation in its winter shoots. The morphological differentiation process of the flower bud differentiation was observed by the paraffin cut section method, flower bud differentiation rate of every period was calculated, the contents changing of total soluble sugar, sucrose, fructose, reducing sugar and starch in buds and leaves were determined, and the contents of soluble protein and the activity changes of a-AMY and POD in matured leaves and young leaves were determined, and analyzed the graphs of soluble protein by SDS—PAGE electrophoresis, a-AMY isozyme and POD isozyme. The study, in the view of comparison theory, explores the relationship between flower buds differentiation and the change of nutrition dynamic in different organs. The analysis indicates that:
1. Process of flower bud development can be divided into 5 stages which are non-differentiation, inflorescence primordial differentiation, main axle of inflorescence differentiation, multistage side inflorescence fast differentiation and colored organ differentiation. During the flower bud differentiation, apical bud was in intermediate states between hibernation or flower bud and leaf bud, which could not be a sign of beginning of flower bud differentiation, but at the beginning of red point expose, the base axillary bud was the first to get into flower bud differentiation than apical bud, and which had obvious staged in differentiation, so flower bud differentiation should subject to morphology differentiation of axillary bud.
2. During the flower bud differentiation, the change trends of soluble sugar content in leaves and buds was "low-high-low", but the change width of content in buds was larger, and content was higher than leaves. The changing trend of starch content in leaves was reduced markedly, while in contrary for buds, which reflected the relationship of the buds and leaves was sink and source respectively during the flower bud differentiation. The correlation of the changing magnitude of total soluble sugar, sucrose and reducing sugar in buds and the flower bud differentiation process was higher compared with leaves, while the correlation of the starch content changes in buds and leaves and the flower bud differentiation process was no significance, which indicated the higher soluble sugar content in buds was to the benefit of flower bud differentiation, but the accumulation level of starch was not determining factor of flowering.
3. During the flower bud differentiation, the protein content, the a-AMY activity and POD activity in matured leaves were higher than young leaves, which had obvious change during flowering period, which indicated those factors proved to be significant to flowering. There were fixed protein bands in protein analysis by SDS-PAGE electrophoresis, which were 220KD,80KD and 55KD, it appeared clearer specific protein bands in main axle of inflorescence differentiation and colored organ differentiation, which was 30KD and 25KD.a-AMY isozyme bands presented gradually brightening. POD isozyme presented new bands in beginning period and inflorescence primordial differentiation, which could be a sign of beginning. Those differences of isozyme expressive content caused the changes of activity, which indicated SDS-PAGE electrophoresis and PAGE electrophoresis were reliable in the research of sijihua longan's flower bud differentiation, but the related question was still worth researching deeply.
本试验以5-6a的四季花龙眼为试材,研究了冬梢花芽分化进程中碳素营养和相关生理活性的变化动态。采用石蜡切片法显微观察其花芽的形态分化,统计各分化时期的花芽分化率,测定其芽体和叶片的可溶性总糖、蔗糖、果糖、还原糖、淀粉和蛋白质含量及淀粉酶和过氧化物酶活性,分析了可溶性蛋白PAGE图谱、淀粉酶同工酶谱和过氧化物酶同工酶谱。从对比的角度探讨了不同器官营养物质变化与花芽分化的关系。结果表明：
1、四季花龙眼花芽形态分化过程可划分为未分化时期、花序原基分化期、花序主轴分化期、多级侧花序快速分化期和花器官分化期。在整个花芽分化过程中,顶芽多处于休眠或花芽和叶芽的中间状态,不能作为花芽分化起始的标志,而基部腋芽从露红点开始,较顶芽率先进入花芽分化期,且各分化时期有明显的阶段性,所以花芽分化应以腋芽形态分化为准。
2、在花芽分化期,叶片和芽体的各种可溶性糖含量的变化均呈现“低-高-低”的变化趋势,但芽体含量的变化幅度较大,其含量也高于叶片。叶片内淀粉含量变化呈显著的降低趋势而芽体内淀粉含量则呈上升趋势。与叶片相比较,芽体中的可溶性总糖、还原糖和蔗糖的含量变化与花芽分化的相关度更高,而芽体和叶片的淀粉含量与花芽分化率无显著相关性,说明芽体内较高的可溶性糖有利于花芽分化的进行,而淀粉积累水平并非成花的决定性因素。
3、成花过程中,成熟叶片的蛋白质含量,α-淀粉酶(α-AMY)活性和过氧化物酶(POD)活性均高于嫩叶含量,并在成花期有明显的变化规律,说明这些因素在成花过程中起到一定的积极意义。可溶性蛋白的SDS-PAGE凝胶电泳分析呈现有固定蛋白条带220KD,80KD和55KD,在花序主轴分化期和花器官快速分化期出现较为清晰的特异蛋白条带30KD和25KD。α-AMY同工酶酶谱呈现出逐渐增亮的条带。POD同工酶酶谱在花芽分化初期和花序原基分化期出现了新的条带,可作为分化的起始标志。它们的酶表达量的差异引起了酶活性的变化,说明SDS-PAGE和PAGE凝胶电泳呈现的条带变化与四季花龙眼花芽分化进程有密切的关系。
Sijihua longan has many advantages which are the habit of anniversary Blossoming, high fruit set, fleshy fruits with flavor of honey and so on. Its many aspects are superior to regular varieties, which include tree vigor Opening, tree stunting and the plant with high density. So to research the process and impact physiologic factors of flower bud differentiation is foundation of reasonable regulate flower bud differentiation process and improve the cultivation benefits.
Five to six-year-old sijihua longan were used as the experimental materials to research the change of nutrition dynamic during flower bud differentiation in its winter shoots. The morphological differentiation process of the flower bud differentiation was observed by the paraffin cut section method, flower bud differentiation rate of every period was calculated, the contents changing of total soluble sugar, sucrose, fructose, reducing sugar and starch in buds and leaves were determined, and the contents of soluble protein and the activity changes of a-AMY and POD in matured leaves and young leaves were determined, and analyzed the graphs of soluble protein by SDS—PAGE electrophoresis, a-AMY isozyme and POD isozyme. The study, in the view of comparison theory, explores the relationship between flower buds differentiation and the change of nutrition dynamic in different organs. The analysis indicates that:
1. Process of flower bud development can be divided into 5 stages which are non-differentiation, inflorescence primordial differentiation, main axle of inflorescence differentiation, multistage side inflorescence fast differentiation and colored organ differentiation. During the flower bud differentiation, apical bud was in intermediate states between hibernation or flower bud and leaf bud, which could not be a sign of beginning of flower bud differentiation, but at the beginning of red point expose, the base axillary bud was the first to get into flower bud differentiation than apical bud, and which had obvious staged in differentiation, so flower bud differentiation should subject to morphology differentiation of axillary bud.
2. During the flower bud differentiation, the change trends of soluble sugar content in leaves and buds was "low-high-low", but the change width of content in buds was larger, and content was higher than leaves. The changing trend of starch content in leaves was reduced markedly, while in contrary for buds, which reflected the relationship of the buds and leaves was sink and source respectively during the flower bud differentiation. The correlation of the changing magnitude of total soluble sugar, sucrose and reducing sugar in buds and the flower bud differentiation process was higher compared with leaves, while the correlation of the starch content changes in buds and leaves and the flower bud differentiation process was no significance, which indicated the higher soluble sugar content in buds was to the benefit of flower bud differentiation, but the accumulation level of starch was not determining factor of flowering.
3. During the flower bud differentiation, the protein content, the a-AMY activity and POD activity in matured leaves were higher than young leaves, which had obvious change during flowering period, which indicated those factors proved to be significant to flowering. There were fixed protein bands in protein analysis by SDS-PAGE electrophoresis, which were 220KD,80KD and 55KD, it appeared clearer specific protein bands in main axle of inflorescence differentiation and colored organ differentiation, which was 30KD and 25KD.a-AMY isozyme bands presented gradually brightening. POD isozyme presented new bands in beginning period and inflorescence primordial differentiation, which could be a sign of beginning. Those differences of isozyme expressive content caused the changes of activity, which indicated SDS-PAGE electrophoresis and PAGE electrophoresis were reliable in the research of sijihua longan's flower bud differentiation, but the related question was still worth researching deeply.
引文
[1]彭坚,李永红,席嘉宾,等.化学调控四季花龙眼成花坐果的初步研究[J].西北植物学报,2005,25(7)：1440-1445.
[2]彭坚,李永红,李论,等.龙眼新品种四季花的选育[J].中国果树,2006,115(5)：28-30.
[3]俞征瑶,彭坚,赵长增,等.‘四季花’龙眼花序二次开花调控的初步研究[J].安徽农业科学,2009,37(7)：2903-2905.
[4]钟伟,林晓东,朱芳德,等.应用RAPD技术分析热带龙眼四季蜜与常规龙眼品种的遗传差异[J].中山大学学报(自然科学版),2004,S1：65-69.
[5]郑丽霞,林晓东,朱芳德,等.龙眼FLO/LFY同源基因cDNA片段的克隆[J].中山大学学报(自然科学版),2004,43(z1)：32-36.
[6]朱芳德,张海岚,吴定尧,等.物理调控促进四季蜜龙眼批量成花座果技术研究[J].广东农业科学,2003,2：45-47.
[7]钟耀兴.四季龙眼越冬果实套袋保果田间试验[J].广西热带农业,2004,3：65-67.
[8]廖明安.园艺植物研究法[M].北京：中国农业出版社,2005：219-236.
[9]河北农业大学.果树栽培学总论[M].北京：农业出版社,1992：59-75.
[10]都荣庭.果树栽培学总论[M].第三版.北京：中国农业出版社,2006：63-71.
[11]吕柳新,林顺权.果树生殖学导论[M].北京：中国农业出版社,1995：25-37.
[12]沈惠娟,黄做喜,桂仁意,等.麝香百合,仙客来,瓜叶菊花芽分化的调控[J].南京林业大学学报,2001,25(2)：55-57.
[13]吴邦良,夏春森,赵宗方,等.果树开花结实生理和调控技术[M].北京：科学技术出版社,1995：25-30.
[14]曹尚银,张秋明,吴顺.果树花芽分化机理研究进展[J].果树学报,2003,20(5)：345-350.
[15]马月萍,戴思兰.植物花芽分化机理研究进展[J].分子植物育种,2003,1(4)：539-545.
[16]李兴军,杨映根,郑文菊,等.果树花芽孕育的研究概况[J].植物学通报,2002,19(4)：385-395.
[17]李文庆,张民,束怀瑞.氮素在果树上的生理作用[J].山东农业大学学报,2002,33(1)：96-100.
[18]刘孝仲,赖毅,许生杰,等.伏令夏橙花芽分化期蛋白质及氨基酸含量的变化[J].园艺学报,1984,11(2)：85-92.
[19]孙文全.多胺代谢与园艺植物开花的关系[J].园艺学报,1989,16(8)：178-183.
[20]杨义标,邵志芳,邱少松,等.中国农学通报[J], 2008,2(24)：281-284.
[21]吴志祥.气象因素对荔枝花芽分化的影响研究[D].儋州：华南热带农业大学,2005：39.
[22]黄蓉.园林植物开花生理与控制[M].农业出版社,1990：30-42.
[23]都荣庭.果树栽培学总论[M].第三版.北京：中国农业出版社,2006：63-71.
[24]吕柳新,林顺权.果树生殖学导论[M].北京：中国农业出版社,1995：25-37.
[25]宁正祥.果树生理调控学[M].广州：华南理工大学出版社,2001：82-126.
[26]郭金丽,张玉兰.苹果梨花芽分化期蛋白质、淀粉代谢的研究[J].内蒙古农牧学院学报,1999,20(2)：80-82.
[27]刘浩荣,宋海星,等.油菜茎叶可溶性糖与游离氨基酸含量的动态变化[J].西北农业学报,2007,16(1)：123-126.
[28]赵江涛,李晓峰,李航等.可溶性糖在高等植物代谢调节中的生理作用[J].安徽农业科学,2006,34(24)：6423-6427.
[29]Koch K E,Ying Z,Wu Y, et al. Multiple paths of sugar-sensing and a sugar/oxygen overlap for genes of sucrose and ethanol metabolism [J].J Exp Bot,2000(51):417-427.
[30]Arenas-Huertero F,Arroyo-Becerra A, Zhou L,et al. Analysis of Arabidopsis glucose insensitive mutants,gin5 and gin6,reveals a central role of the plant hormone ABA in the regulation of plant vegetative development by sugar[J].Gene Dev,2000(14):2085-2096.
[31]Zhou L, Jang J C, Jones T L, et al. Glucose and ethylene signal transduction crosstalk revealed by an arabidopsis glucose insensitive mutant [J]. Proc Natl Acad Sci USA,1998(95): 10294-10299.
[32]Takeda S, Mano S, Ohto M, et al. Inhibitors of protein phosphates 1 and 2A block the sugar-inducible gene expression in plants [J]. PlantPhysiol,1994(106):567-574.
[33]Ohto M A, Hayashi K, Isobe M, et al. Involvement of Ca2+ signaling in the sugar-inducible expression of genes coding forsporamin and β-amylase of sweet potato[J]. Plant,1995(7): 297-307.
[34]樊卫国,刘国琴,安华明,等.刺梨花芽分化期芽中内源激素和碳、氮营养的含量动态[J].果树学报,2003,20(1)：40-43.
[35]罗羽洧,解卫华,马凯.植物营养,核酸与果树花芽分化的关系[J].北方园艺,2007(5)：72-74.
[36]李兴军,李三玉,汪国云,等.杨梅花芽孕育期间叶片酸性蔗糖酶活性及糖类含量的变化[J].四川农业大学学报,2000,18(2)：164-166.
[37]梁芳,郑成淑,张翠华,等.菊花花芽分化过程中芽和叶片碳水化合物含量的变化[J].山东农业科学,2008,1：40-42.
[38]路苹,郭蕊,于同泉,等.切花百合鳞茎花芽形态分化期碳水化合物代谢变化[J].北京农学院学报,2003(4)：259-261.
[39]陈清西,李松刚.KClO3诱导龙眼成花及其叶片碳水化合物与蛋白质的变化[J].福建农业大学学报(自然科学版),2004,33(2)：182-185.
[40]汪炳良,邓俭英,曾广文.萝卜花芽分化过程中茎尖和叶片碳水化合物含量的变化[J].园艺学报,2004,31(3)：375-377.
[41]周家容,周俊辉.宽皮柑桔隔年结果与碳水化合物的关系[J].西南农业大学学报(自然科学版),2004,26(2)：191-197.
[42]李倩,肖建忠,李志斌,等.高山杜鹃花芽分化期生理生化研究[J].河北农业大学,2009,32(1)：47-50.
[43]吴月燕,李培民,吴秋峰.葡萄叶片内碳水化合物及蛋白质代谢对花芽分化的影响[J].浙江万里学院学报,2002,15(4)：54-57.
[44]胡军.枇杷花芽分化和某些物质代谢关系的研究[J].浙江农业大学学报,1983,9(4)：307-314.
[45]何开跃,谢寅峰,艾畅,等.矮牵牛开花过程中核酸和叶绿素含量的变化[J].南京林业大学学报：自然科学版,2004,28(6)：95-97.
[46]潘瑞炽,董愚得.植物生理学[M].北京：高等教育出版社,2001：55-136.
[47]河北农业大学.果树栽培学[M].北京：中国农业出版社出版,1984.
[48]孙旭武,李唯,王力荣,等.桃花芽分化期蛋白质,氨基酸和碳水化合物含量的变化[J].甘肃农业大学学报,2004,39(3)：295-299.
[49]刘宗莉,林顺权,陈厚彬.枇杷成花过程叶片蛋白质变化动态[J].亚热带植物科学,2008,37(2)：1-6.
[50]余沛涛,浦冬华,周伟明.矮牵牛开花过程中的可溶性蛋白质分析[J].植物生理与分子生物学学报,2004,30(2)：179-182.
[51]Monselise S P, Goren R. The role of internal factors and exogenous control in flowering peel growth and abscission in Citrus[J]. Hort science,1978,13(2):134-139.
[52]Banno K, Hayashi S, Tanabe K. Relationships between flower bud formation and nitrogen nutrition in Japanese pear(Pyrus serotina Rehd)[J]. Japan Soc Hort Sci,1984,53(3):265-270.
[53]邓烈,李学柱,何绍兰.柑桔花芽分化与内源激素及淀粉酶活性的关系[J].西南农业大学学报,1991,13(1)：87-91.
[54]何绍兰,李学柱,邓烈.温州蜜柑大小年结果和喷布赤霉酸对花芽分化及过氧化物酶同工酶的影响[J].中国柑桔,1989,18(2)：16-18.
[55]崔明昆,王跃华,马绍兵.同工酶分析在植物居群生物学研究中的应用.云南教育学院学报.1998,14(2)：68-71.
[56]郭春绒,张述义.稀土元素对不同苗龄玉米过氧化物同工酶的影响[J].山西农业大学学报.1994,14(4)：379-381.
[57]刘昌文,丁志祥,程昌文.柑桔属叶片酯酶同工酶和蛋白质电泳分析[J].西南农业学报.1997,10(4)：70-73.
[58]Philippe D.P. Isozymes(A). Rattazzi MC. Genetics and Evolution[C].New York:Alan R Liss,1983: 193-238.
[59]Soltis D E, Sohis P.S. Isozyme evidence for ancient polyploidy in primitive angiosperms[J]. Syst. Bot.1990,15(2):328-337.
[60]Staub J E, Box J, Meglic V, et al. Comparison of isozyme and random amplified polymorphic DNA for determining intra specific variation in Cucumis [J]. Genet.R.Crop Evol.1997,44:257-269.
[61]张士文.酯酶同工酶与番茄杂种优势关系的研究[J].园艺学报,1987,14(2)：108-113.
[62]候彩霞,沈博礼,刘戈等.镧和铈对小麦幼苗过氧化物酶和淀粉酶活性及其同工酶的影响[J].稀土.1997,18(2)：64-66.
[63]杨立新.中国李品种同工酶酶谱稳定性研究[J].北方园艺2007(1)：17-18.
[64]A. S. Haider, A. R. El-Shanshoury and A. Haider. Variation of storage proteins and isozymes within maize inbred lines[J].Biologic Planetarium,2000,43(2):199-203.
[65]周劲松.同工酶在果树研究中的应用[J].果树科学.1988,5(3)：136-141.
[66]T. Stoilova, N. Cholakova and M. Markova. Variation in seed protein and isoenzyme patterns in cucurbita cultivars[J]. Biologic Planetarium,2006,50(3):450-452.
[67]Woo Jin Jung, Fazli Mabood, Tae Hwan Kimh, et al. Induction of pathogenesis-related proteins during biocontrol of rhizoctonia solani with pseudomonas aureofaciens in soybean (Glycine max L. Merr.) plants [J]. BioControl,2007,52:895-904.
[68]N. Mirali, S. El-Khouri and F. Rizq. Genetic diversity and relationships in some vicia species as determined by SDS-PAGE of seed proteins [J]. Biologic Planetarium,2007,51(4):660-666.
[69]Bolwell G P, Wojtaszek D. Mechanisms for the generation of reactive oxygen species in plant defence abroad perspective [J]. Physiol Mol Plant Pathol,1997,51:347-360.
[70]李红艳,杨继华,薛妙男.沙田柚花柱三种同工酶分析[J].广西师范大学学报(自然科学版).1999,17(4)：81-84.
[71]李金亭,王林嵩,马剑敏等.萝卜贮藏期过氧化物酶活性及同工酶谱研究[J].广西植物.2000(1)：59-63.
[72]Liu Z H, Ger M J. Changes of enzyme activity during pollen germination in maize, and possible evidence of lignin synthesis [J]. Plant physiol.1991,24:329-335.
[73]Miguel A. Peroxides and IAA oxidase activities and peroxidase isoenzymes in the pericarp of seeded and seedless "red-haren" peach fruit [J]. Plant Growth Regul.1992,11:1-6.
[74]陆卫,贾敬芬.谷子耐盐系的几种同工酶的变化及外源ABA对它们的影响[J].西北植物学报.1996,16(4)：337-344.
[75]邵继荣,何清海,张述华.水稻多用恢复系130的七种同工酶研究[J].西南农业学报.1999,12(1)：8-13.
[76]梁艳荣,胡晓红,张颖力,等.植物过氧化物酶生理功能研究进展[J].内蒙古农业大学学报,2003,24(2)：110-113.
[77]Brew baker J.L.1980,植物生理生化译丛[C].
[78]王海延.番茄四个种的过氧化物同功酶分析[J].植物研究,1987,7(4)：133-152.
[79]姜华武,汪琳.四倍体黄花菜胚状体发育过程中过氧化物酶的变化[J].湖北农学院学报,1983,18(1)：89-90.
[80]李世泽,刘藿.小麦α-淀粉酶活性的浅探[J].植物生理学报,2001,11(5)：21-25.
[81]杨红,王锁民.豌豆种子萌发时的含水量对子叶中蛋白酶和淀粉酶活性的影响.西北植物学报.2002,22(5)：1136-1143.
[82]沈隽.果树的矿质营养和施肥[J].园艺学报,1980,7(2)：47-56.
[83]黄承和,朱治强,常春荣,等.香蕉花芽分化期的生物量及其养分含量的变化[J].热带作物学报,2006,27(2)：6-10.
[84]邱雪思,刘国成,吕德国,等.杏花芽分化期叶片内源激素含量的变化[J].安徽农业科学,2006,34(9)：1798-1800.
[85]张万萍,史继孔.银杏雄花芽分化期间内源激素,碳水化合物和矿质营养含量的关系[J].林业科学,2004,40(2)：51-54.
[86]赵文东,孙凌俊,徐静,等.薄膜温室葡萄花芽分化规律的研究[J].果树学报,2006,23(1)：9-12.
[87]张林平,杨建民,白志英,等.大石早生李花芽分化研究[J].河北林果研究,1998,13(2)：174-177.
[88]林加涵.现代生物学实验(上册)[M].北京：高等教育出版社,2000：70-86.
[89]张振清.植物材料中可溶性糖的测定[A].薛应龙.植物生理学实验手册[M].上海：上海科技出版社.1985：134-138.
[90]徐昌杰,陈文峻,陈昆松,等.淀粉含量测定的一种简便方法——碘显色法[J].生物技术,1998,8(2)：42-43.
[91]张宪政.植物生理学实验技术[M].沈阳：辽宁科学技术出版社,1994.
[92]李琳,焦新之.应用蛋白质染色剂考马斯亮蓝G-250测定蛋白质的方法[J].植物生理学通讯,1980(6)：52-54.
[93]AMY活性的测定参考刘小丽,宋保军,侯睿林.测定α-淀粉酶活性的两种方法的比较研究[J].农业科技与信息,2006(9)：36-38.
[94]张仲鸣,崔克明,李举怀.白皮松形成层的活动周期及其多糖贮量和淀粉酶同工酶的变化[J].植物学报,1997,39(10)：926-932.
[95]N.Mirali, S.El-Khouri, F.Rizq. Genetic diversity and relationships in some vicia species as determined by SDS-PAGE of seed proteins [J].Biologia plantarum,2007,51(4):660-666.
[96]Yan Pan, Li Jun Wu, Zeng Liang Yu. Effect of salt and drought stress on antioxidant enzymes activities and SOD isoenzymes of liquorice (Glycyrrhizauralensis Fisch)[J].Plant growth regul,2006,49:157-165.
[97]G.Jyothsnakumari, M.Thippeswamy, G. Veeranagamallaiah, et al. Differential expression of LEA proteins in two genotypes of mulberry under salinity [J]. Biologia plantarum,2009,53(1):145-150.
[98]T.Stoilova, N.Cholakova, M.markova. Variation in seed protein and isoenzyme patterns in cucurbita cultivars [J]. Biologia plantarum,2006,50(3):450-452.
[99]A.B.Mandal, Aparna maiti, Bikash Chowdhury, et al. Isoenzyme markers in varietal identification of banana [J]. In vitro cell. Dev. Biol,2001,37(5):599-604.
[100]邱金淡,吴定尧,张海岚.石硖龙眼花芽分化的研究[J].华南农业大学学报,2001,22(1)：27-30.
[101]Kozlowski T. Carbohydrate sources and sinks in woody plant [J].Bot. Rev 1992,58 (2):107-222.
[102]汪炳良,等.萝卜花芽分化过程中茎尖和叶片碳水化合物含量的变化[J].园艺学报,2004,31(3)：375-377.
[103]曾新萍,等.6-BA+B9对龙眼花芽生理分化期内源多胺含量变化的影响[J].亚热带植物科学2006,35(4)：19-21.
[104]陈清西,等.KClO3诱导龙眼成花及其叶片碳水化合物与蛋白质的变化[J].福建农林大学学报(自然科学版),2004,33(2)：182-185.
[105]王宪泽.生物化学实验技术原理和方法[M].中国农业出版社,2002：1-3.
[106]奥岩松,李式军.大白菜发育过程中可溶性蛋白质的变化[J].中国蔬菜,1997(2):19-21.
[107]袁勤生.现代酶学[M].上海：华东理工大学出版社,2001：133-135.
[108]李秉真,李雄,孙庆林.苹果梨花芽分化期内源激素在芽和叶中分布.内蒙古大学学报(自然科学版),1999,36(6)：741-744.
[109]陈万秋,叶新太,李思光等.称猴桃酷酶同工酶和过氧化物酶同工酶分析及遗传多样性研究[J].南昌大学学报(理科版),2001,25(3)：269-270.
[110]陈正华.木本植物组织培养及其应用[M].北京：高等教育出版社,1986：456-465.
[111]陈启林,陈毓荃,陈静,等.同工酶PAGE凝胶电泳在大白菜品种鉴定中的应用研究[J].西北农业学报,1998,7(2)：94-97.
[112]胡能书,万贤国.同工酶技术及其应用[M].长沙：湖南科学技术出版社,1985：73-77.
[113]罗立新,崔克明,李正理.杜仲形成层活动周期中多糖贮量和淀粉酶同工酶的变化[J].北京大学学报(自然科学版),1996,32(2)：231-238.
[114]张鲁刚,何桂兰.大白菜胞质雄性不育系和保持系进行了同工酶和可溶性蛋白质分析[J].西北农业学报,1998,8(4)：67-70.
[115]李玉红,陈鹏,唐爱均,等.不同含水量菜豆种子老化过程中生理特性的研究[J].园艺学报,2005,32(5)：908-910.
[116]张才喜,李载龙,陈大明.湖北海棠阶段转变过程中几种同工酶变化研究[J].植物学通报,2001,18(1)：100-104.
[2]彭坚,李永红,李论,等.龙眼新品种四季花的选育[J].中国果树,2006,115(5)：28-30.
[3]俞征瑶,彭坚,赵长增,等.‘四季花’龙眼花序二次开花调控的初步研究[J].安徽农业科学,2009,37(7)：2903-2905.
[4]钟伟,林晓东,朱芳德,等.应用RAPD技术分析热带龙眼四季蜜与常规龙眼品种的遗传差异[J].中山大学学报(自然科学版),2004,S1：65-69.
[5]郑丽霞,林晓东,朱芳德,等.龙眼FLO/LFY同源基因cDNA片段的克隆[J].中山大学学报(自然科学版),2004,43(z1)：32-36.
[6]朱芳德,张海岚,吴定尧,等.物理调控促进四季蜜龙眼批量成花座果技术研究[J].广东农业科学,2003,2：45-47.
[7]钟耀兴.四季龙眼越冬果实套袋保果田间试验[J].广西热带农业,2004,3：65-67.
[8]廖明安.园艺植物研究法[M].北京：中国农业出版社,2005：219-236.
[9]河北农业大学.果树栽培学总论[M].北京：农业出版社,1992：59-75.
[10]都荣庭.果树栽培学总论[M].第三版.北京：中国农业出版社,2006：63-71.
[11]吕柳新,林顺权.果树生殖学导论[M].北京：中国农业出版社,1995：25-37.
[12]沈惠娟,黄做喜,桂仁意,等.麝香百合,仙客来,瓜叶菊花芽分化的调控[J].南京林业大学学报,2001,25(2)：55-57.
[13]吴邦良,夏春森,赵宗方,等.果树开花结实生理和调控技术[M].北京：科学技术出版社,1995：25-30.
[14]曹尚银,张秋明,吴顺.果树花芽分化机理研究进展[J].果树学报,2003,20(5)：345-350.
[15]马月萍,戴思兰.植物花芽分化机理研究进展[J].分子植物育种,2003,1(4)：539-545.
[16]李兴军,杨映根,郑文菊,等.果树花芽孕育的研究概况[J].植物学通报,2002,19(4)：385-395.
[17]李文庆,张民,束怀瑞.氮素在果树上的生理作用[J].山东农业大学学报,2002,33(1)：96-100.
[18]刘孝仲,赖毅,许生杰,等.伏令夏橙花芽分化期蛋白质及氨基酸含量的变化[J].园艺学报,1984,11(2)：85-92.
[19]孙文全.多胺代谢与园艺植物开花的关系[J].园艺学报,1989,16(8)：178-183.
[20]杨义标,邵志芳,邱少松,等.中国农学通报[J], 2008,2(24)：281-284.
[21]吴志祥.气象因素对荔枝花芽分化的影响研究[D].儋州：华南热带农业大学,2005：39.
[22]黄蓉.园林植物开花生理与控制[M].农业出版社,1990：30-42.
[23]都荣庭.果树栽培学总论[M].第三版.北京：中国农业出版社,2006：63-71.
[24]吕柳新,林顺权.果树生殖学导论[M].北京：中国农业出版社,1995：25-37.
[25]宁正祥.果树生理调控学[M].广州：华南理工大学出版社,2001：82-126.
[26]郭金丽,张玉兰.苹果梨花芽分化期蛋白质、淀粉代谢的研究[J].内蒙古农牧学院学报,1999,20(2)：80-82.
[27]刘浩荣,宋海星,等.油菜茎叶可溶性糖与游离氨基酸含量的动态变化[J].西北农业学报,2007,16(1)：123-126.
[28]赵江涛,李晓峰,李航等.可溶性糖在高等植物代谢调节中的生理作用[J].安徽农业科学,2006,34(24)：6423-6427.
[29]Koch K E,Ying Z,Wu Y, et al. Multiple paths of sugar-sensing and a sugar/oxygen overlap for genes of sucrose and ethanol metabolism [J].J Exp Bot,2000(51):417-427.
[30]Arenas-Huertero F,Arroyo-Becerra A, Zhou L,et al. Analysis of Arabidopsis glucose insensitive mutants,gin5 and gin6,reveals a central role of the plant hormone ABA in the regulation of plant vegetative development by sugar[J].Gene Dev,2000(14):2085-2096.
[31]Zhou L, Jang J C, Jones T L, et al. Glucose and ethylene signal transduction crosstalk revealed by an arabidopsis glucose insensitive mutant [J]. Proc Natl Acad Sci USA,1998(95): 10294-10299.
[32]Takeda S, Mano S, Ohto M, et al. Inhibitors of protein phosphates 1 and 2A block the sugar-inducible gene expression in plants [J]. PlantPhysiol,1994(106):567-574.
[33]Ohto M A, Hayashi K, Isobe M, et al. Involvement of Ca2+ signaling in the sugar-inducible expression of genes coding forsporamin and β-amylase of sweet potato[J]. Plant,1995(7): 297-307.
[34]樊卫国,刘国琴,安华明,等.刺梨花芽分化期芽中内源激素和碳、氮营养的含量动态[J].果树学报,2003,20(1)：40-43.
[35]罗羽洧,解卫华,马凯.植物营养,核酸与果树花芽分化的关系[J].北方园艺,2007(5)：72-74.
[36]李兴军,李三玉,汪国云,等.杨梅花芽孕育期间叶片酸性蔗糖酶活性及糖类含量的变化[J].四川农业大学学报,2000,18(2)：164-166.
[37]梁芳,郑成淑,张翠华,等.菊花花芽分化过程中芽和叶片碳水化合物含量的变化[J].山东农业科学,2008,1：40-42.
[38]路苹,郭蕊,于同泉,等.切花百合鳞茎花芽形态分化期碳水化合物代谢变化[J].北京农学院学报,2003(4)：259-261.
[39]陈清西,李松刚.KClO3诱导龙眼成花及其叶片碳水化合物与蛋白质的变化[J].福建农业大学学报(自然科学版),2004,33(2)：182-185.
[40]汪炳良,邓俭英,曾广文.萝卜花芽分化过程中茎尖和叶片碳水化合物含量的变化[J].园艺学报,2004,31(3)：375-377.
[41]周家容,周俊辉.宽皮柑桔隔年结果与碳水化合物的关系[J].西南农业大学学报(自然科学版),2004,26(2)：191-197.
[42]李倩,肖建忠,李志斌,等.高山杜鹃花芽分化期生理生化研究[J].河北农业大学,2009,32(1)：47-50.
[43]吴月燕,李培民,吴秋峰.葡萄叶片内碳水化合物及蛋白质代谢对花芽分化的影响[J].浙江万里学院学报,2002,15(4)：54-57.
[44]胡军.枇杷花芽分化和某些物质代谢关系的研究[J].浙江农业大学学报,1983,9(4)：307-314.
[45]何开跃,谢寅峰,艾畅,等.矮牵牛开花过程中核酸和叶绿素含量的变化[J].南京林业大学学报：自然科学版,2004,28(6)：95-97.
[46]潘瑞炽,董愚得.植物生理学[M].北京：高等教育出版社,2001：55-136.
[47]河北农业大学.果树栽培学[M].北京：中国农业出版社出版,1984.
[48]孙旭武,李唯,王力荣,等.桃花芽分化期蛋白质,氨基酸和碳水化合物含量的变化[J].甘肃农业大学学报,2004,39(3)：295-299.
[49]刘宗莉,林顺权,陈厚彬.枇杷成花过程叶片蛋白质变化动态[J].亚热带植物科学,2008,37(2)：1-6.
[50]余沛涛,浦冬华,周伟明.矮牵牛开花过程中的可溶性蛋白质分析[J].植物生理与分子生物学学报,2004,30(2)：179-182.
[51]Monselise S P, Goren R. The role of internal factors and exogenous control in flowering peel growth and abscission in Citrus[J]. Hort science,1978,13(2):134-139.
[52]Banno K, Hayashi S, Tanabe K. Relationships between flower bud formation and nitrogen nutrition in Japanese pear(Pyrus serotina Rehd)[J]. Japan Soc Hort Sci,1984,53(3):265-270.
[53]邓烈,李学柱,何绍兰.柑桔花芽分化与内源激素及淀粉酶活性的关系[J].西南农业大学学报,1991,13(1)：87-91.
[54]何绍兰,李学柱,邓烈.温州蜜柑大小年结果和喷布赤霉酸对花芽分化及过氧化物酶同工酶的影响[J].中国柑桔,1989,18(2)：16-18.
[55]崔明昆,王跃华,马绍兵.同工酶分析在植物居群生物学研究中的应用.云南教育学院学报.1998,14(2)：68-71.
[56]郭春绒,张述义.稀土元素对不同苗龄玉米过氧化物同工酶的影响[J].山西农业大学学报.1994,14(4)：379-381.
[57]刘昌文,丁志祥,程昌文.柑桔属叶片酯酶同工酶和蛋白质电泳分析[J].西南农业学报.1997,10(4)：70-73.
[58]Philippe D.P. Isozymes(A). Rattazzi MC. Genetics and Evolution[C].New York:Alan R Liss,1983: 193-238.
[59]Soltis D E, Sohis P.S. Isozyme evidence for ancient polyploidy in primitive angiosperms[J]. Syst. Bot.1990,15(2):328-337.
[60]Staub J E, Box J, Meglic V, et al. Comparison of isozyme and random amplified polymorphic DNA for determining intra specific variation in Cucumis [J]. Genet.R.Crop Evol.1997,44:257-269.
[61]张士文.酯酶同工酶与番茄杂种优势关系的研究[J].园艺学报,1987,14(2)：108-113.
[62]候彩霞,沈博礼,刘戈等.镧和铈对小麦幼苗过氧化物酶和淀粉酶活性及其同工酶的影响[J].稀土.1997,18(2)：64-66.
[63]杨立新.中国李品种同工酶酶谱稳定性研究[J].北方园艺2007(1)：17-18.
[64]A. S. Haider, A. R. El-Shanshoury and A. Haider. Variation of storage proteins and isozymes within maize inbred lines[J].Biologic Planetarium,2000,43(2):199-203.
[65]周劲松.同工酶在果树研究中的应用[J].果树科学.1988,5(3)：136-141.
[66]T. Stoilova, N. Cholakova and M. Markova. Variation in seed protein and isoenzyme patterns in cucurbita cultivars[J]. Biologic Planetarium,2006,50(3):450-452.
[67]Woo Jin Jung, Fazli Mabood, Tae Hwan Kimh, et al. Induction of pathogenesis-related proteins during biocontrol of rhizoctonia solani with pseudomonas aureofaciens in soybean (Glycine max L. Merr.) plants [J]. BioControl,2007,52:895-904.
[68]N. Mirali, S. El-Khouri and F. Rizq. Genetic diversity and relationships in some vicia species as determined by SDS-PAGE of seed proteins [J]. Biologic Planetarium,2007,51(4):660-666.
[69]Bolwell G P, Wojtaszek D. Mechanisms for the generation of reactive oxygen species in plant defence abroad perspective [J]. Physiol Mol Plant Pathol,1997,51:347-360.
[70]李红艳,杨继华,薛妙男.沙田柚花柱三种同工酶分析[J].广西师范大学学报(自然科学版).1999,17(4)：81-84.
[71]李金亭,王林嵩,马剑敏等.萝卜贮藏期过氧化物酶活性及同工酶谱研究[J].广西植物.2000(1)：59-63.
[72]Liu Z H, Ger M J. Changes of enzyme activity during pollen germination in maize, and possible evidence of lignin synthesis [J]. Plant physiol.1991,24:329-335.
[73]Miguel A. Peroxides and IAA oxidase activities and peroxidase isoenzymes in the pericarp of seeded and seedless "red-haren" peach fruit [J]. Plant Growth Regul.1992,11:1-6.
[74]陆卫,贾敬芬.谷子耐盐系的几种同工酶的变化及外源ABA对它们的影响[J].西北植物学报.1996,16(4)：337-344.
[75]邵继荣,何清海,张述华.水稻多用恢复系130的七种同工酶研究[J].西南农业学报.1999,12(1)：8-13.
[76]梁艳荣,胡晓红,张颖力,等.植物过氧化物酶生理功能研究进展[J].内蒙古农业大学学报,2003,24(2)：110-113.
[77]Brew baker J.L.1980,植物生理生化译丛[C].
[78]王海延.番茄四个种的过氧化物同功酶分析[J].植物研究,1987,7(4)：133-152.
[79]姜华武,汪琳.四倍体黄花菜胚状体发育过程中过氧化物酶的变化[J].湖北农学院学报,1983,18(1)：89-90.
[80]李世泽,刘藿.小麦α-淀粉酶活性的浅探[J].植物生理学报,2001,11(5)：21-25.
[81]杨红,王锁民.豌豆种子萌发时的含水量对子叶中蛋白酶和淀粉酶活性的影响.西北植物学报.2002,22(5)：1136-1143.
[82]沈隽.果树的矿质营养和施肥[J].园艺学报,1980,7(2)：47-56.
[83]黄承和,朱治强,常春荣,等.香蕉花芽分化期的生物量及其养分含量的变化[J].热带作物学报,2006,27(2)：6-10.
[84]邱雪思,刘国成,吕德国,等.杏花芽分化期叶片内源激素含量的变化[J].安徽农业科学,2006,34(9)：1798-1800.
[85]张万萍,史继孔.银杏雄花芽分化期间内源激素,碳水化合物和矿质营养含量的关系[J].林业科学,2004,40(2)：51-54.
[86]赵文东,孙凌俊,徐静,等.薄膜温室葡萄花芽分化规律的研究[J].果树学报,2006,23(1)：9-12.
[87]张林平,杨建民,白志英,等.大石早生李花芽分化研究[J].河北林果研究,1998,13(2)：174-177.
[88]林加涵.现代生物学实验(上册)[M].北京：高等教育出版社,2000：70-86.
[89]张振清.植物材料中可溶性糖的测定[A].薛应龙.植物生理学实验手册[M].上海：上海科技出版社.1985：134-138.
[90]徐昌杰,陈文峻,陈昆松,等.淀粉含量测定的一种简便方法——碘显色法[J].生物技术,1998,8(2)：42-43.
[91]张宪政.植物生理学实验技术[M].沈阳：辽宁科学技术出版社,1994.
[92]李琳,焦新之.应用蛋白质染色剂考马斯亮蓝G-250测定蛋白质的方法[J].植物生理学通讯,1980(6)：52-54.
[93]AMY活性的测定参考刘小丽,宋保军,侯睿林.测定α-淀粉酶活性的两种方法的比较研究[J].农业科技与信息,2006(9)：36-38.
[94]张仲鸣,崔克明,李举怀.白皮松形成层的活动周期及其多糖贮量和淀粉酶同工酶的变化[J].植物学报,1997,39(10)：926-932.
[95]N.Mirali, S.El-Khouri, F.Rizq. Genetic diversity and relationships in some vicia species as determined by SDS-PAGE of seed proteins [J].Biologia plantarum,2007,51(4):660-666.
[96]Yan Pan, Li Jun Wu, Zeng Liang Yu. Effect of salt and drought stress on antioxidant enzymes activities and SOD isoenzymes of liquorice (Glycyrrhizauralensis Fisch)[J].Plant growth regul,2006,49:157-165.
[97]G.Jyothsnakumari, M.Thippeswamy, G. Veeranagamallaiah, et al. Differential expression of LEA proteins in two genotypes of mulberry under salinity [J]. Biologia plantarum,2009,53(1):145-150.
[98]T.Stoilova, N.Cholakova, M.markova. Variation in seed protein and isoenzyme patterns in cucurbita cultivars [J]. Biologia plantarum,2006,50(3):450-452.
[99]A.B.Mandal, Aparna maiti, Bikash Chowdhury, et al. Isoenzyme markers in varietal identification of banana [J]. In vitro cell. Dev. Biol,2001,37(5):599-604.
[100]邱金淡,吴定尧,张海岚.石硖龙眼花芽分化的研究[J].华南农业大学学报,2001,22(1)：27-30.
[101]Kozlowski T. Carbohydrate sources and sinks in woody plant [J].Bot. Rev 1992,58 (2):107-222.
[102]汪炳良,等.萝卜花芽分化过程中茎尖和叶片碳水化合物含量的变化[J].园艺学报,2004,31(3)：375-377.
[103]曾新萍,等.6-BA+B9对龙眼花芽生理分化期内源多胺含量变化的影响[J].亚热带植物科学2006,35(4)：19-21.
[104]陈清西,等.KClO3诱导龙眼成花及其叶片碳水化合物与蛋白质的变化[J].福建农林大学学报(自然科学版),2004,33(2)：182-185.
[105]王宪泽.生物化学实验技术原理和方法[M].中国农业出版社,2002：1-3.
[106]奥岩松,李式军.大白菜发育过程中可溶性蛋白质的变化[J].中国蔬菜,1997(2):19-21.
[107]袁勤生.现代酶学[M].上海：华东理工大学出版社,2001：133-135.
[108]李秉真,李雄,孙庆林.苹果梨花芽分化期内源激素在芽和叶中分布.内蒙古大学学报(自然科学版),1999,36(6)：741-744.
[109]陈万秋,叶新太,李思光等.称猴桃酷酶同工酶和过氧化物酶同工酶分析及遗传多样性研究[J].南昌大学学报(理科版),2001,25(3)：269-270.
[110]陈正华.木本植物组织培养及其应用[M].北京：高等教育出版社,1986：456-465.
[111]陈启林,陈毓荃,陈静,等.同工酶PAGE凝胶电泳在大白菜品种鉴定中的应用研究[J].西北农业学报,1998,7(2)：94-97.
[112]胡能书,万贤国.同工酶技术及其应用[M].长沙：湖南科学技术出版社,1985：73-77.
[113]罗立新,崔克明,李正理.杜仲形成层活动周期中多糖贮量和淀粉酶同工酶的变化[J].北京大学学报(自然科学版),1996,32(2)：231-238.
[114]张鲁刚,何桂兰.大白菜胞质雄性不育系和保持系进行了同工酶和可溶性蛋白质分析[J].西北农业学报,1998,8(4)：67-70.
[115]李玉红,陈鹏,唐爱均,等.不同含水量菜豆种子老化过程中生理特性的研究[J].园艺学报,2005,32(5)：908-910.
[116]张才喜,李载龙,陈大明.湖北海棠阶段转变过程中几种同工酶变化研究[J].植物学通报,2001,18(1)：100-104.