甘蓝型黄籽油菜种皮特性形成机理研究
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摘要
在相同的遗传背景下，甘蓝型油菜(Brassicanapus L)黄籽与黑籽相比黄籽具有含油量高、蛋白质含量高、皮壳率低、油质清澈透明、饼粕饲用价值高等一系列优点，受到油菜育种家的普遍重视。但甘蓝型油菜黄籽性状遗传不稳定，大大制约了黄籽油菜的育种进程，弄清甘蓝型黄籽油菜种皮特性的形成机理，将对黄籽油菜的育种具有重要意义。本文以3对不同来源的甘蓝型油菜作为试验材料，每对材料包括具有近等基因系的黄籽和黑籽两种材料，对甘蓝型黄籽油菜种皮特性形成的机理进行了研究，主要结果如下：
1．种子发育进程中种皮所含色素的动态研究和方差分析表明，遗传背景对种皮色素含量的影响不大。种皮叶绿素、多酚、花色素、黑色素含量在黄、黑籽之间均存在极显著差异：叶绿素含量在种子发育后期黄籽极显著高于黑籽；多酚、花色素、黑色素在黑籽中的含量极显著地高于黄籽：在黑籽种皮颜色不断加深的过程中，多酚、花色素含量有明显的下降，而黑色素含量却迅速上升。认为叶绿素不是产生黑籽种皮色泽的直接原因，多酚、花色素含量对种皮色泽的形成有重要影响，而黑色素含量是决定成熟种皮色泽的主要因素。
2种子发育进程中种皮厚度相关指标的动态研究和方差分析表明，遗传背景对种皮纤维素含量有极显著的影响，对皮壳率及种皮木质素含量的影响却不大。在种子发育进程中，种皮纤维素含量在黄、黑籽间的差异不显著，而皮壳率及种皮木质素含量在黄、黑籽间的差异达显著水平。认为造成黄籽皮壳率低的原因可能是黄籽种皮木质素含量明显低于黑籽所致，而不是纤维素含量的差异造成的。
3．种子发育进程中与种皮色素及木质素合成相关的酶活性的动态研究和方差分析表明，PAL、PPO、POD在种子发育进程中酶活性均是先上升再下降，黑籽的PAL、PPO及POD活性均极显著地高于黄籽，说明PAL、PPO、POD活性对种皮特性的形成均有重要影响。其中，PPO及POD的活性高峰出现在开花后41天左右，PAL活性高峰出现在开花后34天左右，认为PPO及POD在种子呈色过程中可能起了重要作用，而PAL对种皮特性形成的影响，不是直接产生颜色物质或使种皮加厚的物质，而是以产生前体物质为主。
4．以多种酶抑制剂和蛋白质合成调节剂在种子发育进程中处理植株，并对处理植株成熟种子的相关指标进行测定，结果表明，抑制PAL、PPO活性，促进蛋白质合成、抑制氧化反应，均可使种皮色素及木质素含量降低，皮壳率下降、蛋白质含量和含油量有所上升；抑制蛋白质合成使种皮色素及木质素含量有所上升，蛋白质含量、含油量有所下降。说明甘蓝型黄籽油菜种皮特性的形成与莽草酸——苯丙氨酸途径有关，受到PAL、PPO活性及体内氧化反应的影响，这与
甘鱼型黄籽必莱升友甘佐形戍机o研允 戈要
动态测定的结果是相符的。
5．对酯酶、PPO、POD的同工酶分析表明，在相同的遗传背景下，酯酶的同
工酶数在黄、黑籽间没有差异；黑籽的PPO同工酶带均比黄籽多，且黑籽具有1
条虹值为0．41的特异谱带＿带；黑籽的POD同工酶均比黄籽多一条谱带伪
带或。带）。在不同品种间，酯酶、PPO、POD的同工酶谱差异均较大。可能黄
籽缺失的某些同工酶对种皮特性的形成产生了影响。
以上2、3、4、5结论在甘蓝型黄籽油菜的研究中均属首次报道。
Yellow seeds in Brassica species are characterized with thinner seedcoat and lower hull percentage which in turn are correlated with higher oil and protein and lower crude fiber contents compared to black seeds under the same genetic conditions. But the widespread application of yellow-seeded species in Brassica napus L. is limited seriously owing to it's genetic instability.Therefore,the search of the mechanism on the formation of distinctive seedcoat features is meaningful for both rapeseed breeding and plant genetics.
Many studies were on the pigments in the seedcoatUp to now, rare researches were reported of the reason on the lower hull percentage in yellow-seeded rape.Three pairs of different genetic near-isogenic lines of black- and yellow-seeded Brassica napus as experimental materials were used in my study. The results are as follows:
1 .The changes in pigments in seedcoat were investigated with the development of rape seed.The results of experiments and variance analysis showed that there was no significant difference in pigments contents among the three pairs of materials.Butjthe contents of chlorophyll,polyphenols,anthocyanin and melanin in seedcoat were different between black- and yellow-seeded lines and the differences were up to highly significant level. The results showed that chlorophyll content in yellow-seeded lines was much higher than that in black ones when seeds are nearing mature.The contents of polyphenols,anthocyanin and melanin in seedcoat of black-seeded lines were significantly higher than those of yellow-seeded ones.The contents of polyphenols and anthocyanin declined and the content of melanin increased rapidly in the late stage of seed development. So it was speculated that polyphenols and anthocyanin contents might influence the colour of seedcoat,but melanin content might be the mam factor to influence the colour of seedcoat.
2.The indexes related to thickness of seedcoat were measured with the seed
growth.The results of experiments and variance analysis showed that the difference of cellulose content in seedcoat reached highly significant level, but the content of lignin and hull percentage nearly had no difference among the three pairs of materials.The difference in cellulose content between yellow- and black-seeded lines was not obvious during the growth of seed,but the differences in lignin content and hull percentage was up to highly significant level.So it could be concluded that the lower lignin content might be the reason for the lower hull percentage in yellow-seeded lines.
3.some enzymes related to pigments and lignin in seedcoat were studied.The results of experiments and variance analysis showed that the activities of PAL,PPO,POD increased gradually in the early stage and decreased in late stage during the period of seed development.The activities of PAL,PPO,POD in black-seeded lines were much higher than those in yellow ones.lt was suggested that the activities of PAL,PPO and POD might have great effect on the formation of distinctive seedcoat features.The activities of PPO and POD reached climax at 41 DAF.The activity of PAL reached climax at 34 DAF.lt was indicated that PAL activity might mainly influence the content of some precursor.
4.Some enzyme activities were repressed and formation of protein was modulated during the seed growth.The results indicated that the formation of distinctive seedcoat features was related to the shikimic-phenylalanine pathway and influenced by PAL activity,PPO activity and oxidation.
5.The isozyme patterns of esterase,polyphenol oxidase(PPO) and peroxidase(POD) in seedcoat were investigated. Same or similar esterase isozyme bands were observed between black- and yellow-seeded lines under same genetic conditions;There were more PPO isozyme bands in black-seeded lines than in yellow ones and there was 1 special band in black-seeded lines;Black-seeded lines had 1 more POD isozyme band (b or c band) than yellow ones under same genetic conditions. Owing to different genetic background,the isozyme patterns of esterase.PPO an
1．种子发育进程中种皮所含色素的动态研究和方差分析表明，遗传背景对种皮色素含量的影响不大。种皮叶绿素、多酚、花色素、黑色素含量在黄、黑籽之间均存在极显著差异：叶绿素含量在种子发育后期黄籽极显著高于黑籽；多酚、花色素、黑色素在黑籽中的含量极显著地高于黄籽：在黑籽种皮颜色不断加深的过程中，多酚、花色素含量有明显的下降，而黑色素含量却迅速上升。认为叶绿素不是产生黑籽种皮色泽的直接原因，多酚、花色素含量对种皮色泽的形成有重要影响，而黑色素含量是决定成熟种皮色泽的主要因素。
2种子发育进程中种皮厚度相关指标的动态研究和方差分析表明，遗传背景对种皮纤维素含量有极显著的影响，对皮壳率及种皮木质素含量的影响却不大。在种子发育进程中，种皮纤维素含量在黄、黑籽间的差异不显著，而皮壳率及种皮木质素含量在黄、黑籽间的差异达显著水平。认为造成黄籽皮壳率低的原因可能是黄籽种皮木质素含量明显低于黑籽所致，而不是纤维素含量的差异造成的。
3．种子发育进程中与种皮色素及木质素合成相关的酶活性的动态研究和方差分析表明，PAL、PPO、POD在种子发育进程中酶活性均是先上升再下降，黑籽的PAL、PPO及POD活性均极显著地高于黄籽，说明PAL、PPO、POD活性对种皮特性的形成均有重要影响。其中，PPO及POD的活性高峰出现在开花后41天左右，PAL活性高峰出现在开花后34天左右，认为PPO及POD在种子呈色过程中可能起了重要作用，而PAL对种皮特性形成的影响，不是直接产生颜色物质或使种皮加厚的物质，而是以产生前体物质为主。
4．以多种酶抑制剂和蛋白质合成调节剂在种子发育进程中处理植株，并对处理植株成熟种子的相关指标进行测定，结果表明，抑制PAL、PPO活性，促进蛋白质合成、抑制氧化反应，均可使种皮色素及木质素含量降低，皮壳率下降、蛋白质含量和含油量有所上升；抑制蛋白质合成使种皮色素及木质素含量有所上升，蛋白质含量、含油量有所下降。说明甘蓝型黄籽油菜种皮特性的形成与莽草酸——苯丙氨酸途径有关，受到PAL、PPO活性及体内氧化反应的影响，这与
甘鱼型黄籽必莱升友甘佐形戍机o研允 戈要
动态测定的结果是相符的。
5．对酯酶、PPO、POD的同工酶分析表明，在相同的遗传背景下，酯酶的同
工酶数在黄、黑籽间没有差异；黑籽的PPO同工酶带均比黄籽多，且黑籽具有1
条虹值为0．41的特异谱带＿带；黑籽的POD同工酶均比黄籽多一条谱带伪
带或。带）。在不同品种间，酯酶、PPO、POD的同工酶谱差异均较大。可能黄
籽缺失的某些同工酶对种皮特性的形成产生了影响。
以上2、3、4、5结论在甘蓝型黄籽油菜的研究中均属首次报道。
Yellow seeds in Brassica species are characterized with thinner seedcoat and lower hull percentage which in turn are correlated with higher oil and protein and lower crude fiber contents compared to black seeds under the same genetic conditions. But the widespread application of yellow-seeded species in Brassica napus L. is limited seriously owing to it's genetic instability.Therefore,the search of the mechanism on the formation of distinctive seedcoat features is meaningful for both rapeseed breeding and plant genetics.
Many studies were on the pigments in the seedcoatUp to now, rare researches were reported of the reason on the lower hull percentage in yellow-seeded rape.Three pairs of different genetic near-isogenic lines of black- and yellow-seeded Brassica napus as experimental materials were used in my study. The results are as follows:
1 .The changes in pigments in seedcoat were investigated with the development of rape seed.The results of experiments and variance analysis showed that there was no significant difference in pigments contents among the three pairs of materials.Butjthe contents of chlorophyll,polyphenols,anthocyanin and melanin in seedcoat were different between black- and yellow-seeded lines and the differences were up to highly significant level. The results showed that chlorophyll content in yellow-seeded lines was much higher than that in black ones when seeds are nearing mature.The contents of polyphenols,anthocyanin and melanin in seedcoat of black-seeded lines were significantly higher than those of yellow-seeded ones.The contents of polyphenols and anthocyanin declined and the content of melanin increased rapidly in the late stage of seed development. So it was speculated that polyphenols and anthocyanin contents might influence the colour of seedcoat,but melanin content might be the mam factor to influence the colour of seedcoat.
2.The indexes related to thickness of seedcoat were measured with the seed
growth.The results of experiments and variance analysis showed that the difference of cellulose content in seedcoat reached highly significant level, but the content of lignin and hull percentage nearly had no difference among the three pairs of materials.The difference in cellulose content between yellow- and black-seeded lines was not obvious during the growth of seed,but the differences in lignin content and hull percentage was up to highly significant level.So it could be concluded that the lower lignin content might be the reason for the lower hull percentage in yellow-seeded lines.
3.some enzymes related to pigments and lignin in seedcoat were studied.The results of experiments and variance analysis showed that the activities of PAL,PPO,POD increased gradually in the early stage and decreased in late stage during the period of seed development.The activities of PAL,PPO,POD in black-seeded lines were much higher than those in yellow ones.lt was suggested that the activities of PAL,PPO and POD might have great effect on the formation of distinctive seedcoat features.The activities of PPO and POD reached climax at 41 DAF.The activity of PAL reached climax at 34 DAF.lt was indicated that PAL activity might mainly influence the content of some precursor.
4.Some enzyme activities were repressed and formation of protein was modulated during the seed growth.The results indicated that the formation of distinctive seedcoat features was related to the shikimic-phenylalanine pathway and influenced by PAL activity,PPO activity and oxidation.
5.The isozyme patterns of esterase,polyphenol oxidase(PPO) and peroxidase(POD) in seedcoat were investigated. Same or similar esterase isozyme bands were observed between black- and yellow-seeded lines under same genetic conditions;There were more PPO isozyme bands in black-seeded lines than in yellow ones and there was 1 special band in black-seeded lines;Black-seeded lines had 1 more POD isozyme band (b or c band) than yellow ones under same genetic conditions. Owing to different genetic background,the isozyme patterns of esterase.PPO an
引文
马峙英，刘叔倩，王省芬等．过氧化物酶同工酶与棉花黄萎病抗性的相关研究．作物学报．2000，26(4)：431～437
王三根主编．植物生理研究技术．西南农业大学．2000
王汉中，刘后利．油菜皮壳中多酚氧化酶活性的组织化学鉴定及其与粒色的关系．中国油料，1991，1：30～32
王汉中，刘后利．甘蓝型油菜粒色与种子有关性状间的关系研究．中国油料，1989，11(2)：32～35
王保通，商鸿生．小麦高温抗条锈性表达与木质素合成的关系植物保护学报，1996，23(3)：229～233
王敬文，薛应龙．苯丙氨酸解氨酶在抗马铃薯晚疫病中的作用．植物生理学报，1982，8(1)：35～43
王岩，刘学惠，尹佩玉等．几种天然黑色素分子结构的红外光谱表征研究．分析试验室，1996，15(6)：63～65
王新平，李加新．黑糯米中黑色素提取方法的研究．食品科学，1997，18(2)：44～46
田国忠，李怀方，裘维藩．植物过氧化物酶研究进展．武汉植物学研究，2001，19(4)：332～344
丛斌，杨茂成，粟波等．小麦根尖细胞分化过程中木质素合成及其相关酶的活性变化．复旦学报(自然科学版)，1997，36(5)：550～554
白宝璋，汤学军主编．植物生理学测试技术．中国科学技术出版社，1993
仲飞．红星苹果多酚氧化酶某些特性及其抑制剂的研究．园艺学报，1998，25(2)：184～186
朱广廉等．植物生理学试验．北京大学出版社，1990
朱利泉主编．基础生物化学试验原理与方法．成都科技大学出版社，1993
池方，李树人，田红星．红光对套袋苹果花青素含量的影响．河南农业大学学报，1997，31(2)：174～177
刘后利主编．实用油菜栽培学．上海科学出版社．1987，559～561
李加纳，张学昆，谌利等．不同遗传背景的甘蓝型黄籽油菜粒色遗传初步研究．中国油料作物学报，1998，20(4)：16～19
李崇辉，陈文艺．甘蓝型油菜黄籽分布与种皮颜色变化研究．西南农业大学学报．1998，20(3)：256～259
胡能书等．同工酶技术及应用．湖南科学技术出版社，1985
胡晓君．不同类型油菜品种种皮色素含量的关系的研究(硕士论文)．华中农业大学，1988
张小军，牟方生，黄观武等．棉花生化育种技术的初步研究．中国棉花，1996，23(8)：7～8
张健军，李祥，侯明生．小麦品种对梭条斑花叶病毒病抗病特性的研究．Ⅱ抗病性与植株内多酚氧化酶活性及同工酶酶谱的关系．华中农业大学学报，1996，15(5)：420～425
陈玉萍，高永同，刘后利．甘蓝型黄籽油菜粒色及主要品质性状的动态研究．中国油料，1987，4：10～14
陈玉萍，刘后利．甘蓝型油菜种子油菜种子发育进程中种皮色素含量动态．中国油料，1994，16(4)：13～16
陈玉萍，刘后利．甘蓝型油菜种子油菜种子发育进程中种皮颜色变化．中国油料，1995，17(2)：1～3
陈秀芳，王坤范．桃果实发育中褐变因子变化规律的研究．园艺学报，1995，22(3)：230～234
陈炜，刘维营，雷清等．三粒小麦和普通小麦苗期过氧化物酶同工酶和蛋白质的比较研究．作物学报，1999，25(5)：650～653
陈健中，盛炳成，刘克均．苯丙酸代谢与苹果对轮纹病抗性的关系．果树科学，1996，14(3)：149～152
吴江生，石淑稳，刘后利．甘蓝型油菜黄籽突变体的遗传研究．中国油料作物学报，1998，20(3)：5～8
吴振先，苏美霞，陈维信．贮藏荔枝果皮多酚氧化酶及过氧化物酶与褐变的研究．华南农业大学学报，1998，19(1)：12～15
吴耕西，周宏伟，王健民．鸭梨酶促褐变的生化机制及底物鉴定．园艺学报，1992，19(3)：198～202
余沛涛，薛应龙．植物苯丙氨酸解氨酶的研究．植物生理学报，1983，9(3)：301～306
周旭章，魏开华，陈朝辉等．从黑芝麻中提取黑色素的研究．林产化学通讯，1997，4：17～19
祝军，生吉萍，申林等．苹果花青素形成与PAL活性及蛋白质含量的关系．中国农业大学学报，1997，2(3)：97～99
姚金宝，许才康．甘蓝型黄籽油菜种子油菜种子发育进程中若干特性的研究．中国油料，1995，17(3)：4～6
姜慧芳，孙大容，段乃雄等．利用酯酶同工酶分析花生属种间亲缘关系．中国油料，1989，4：21～23
袁艺．侧耳(平菇)的酯酶、多酚氧化酶及过氧化物酶同工酶的电泳分析．安徽农业大学学报，1997，24(1)：88～92
宴本菊，李焕秀．梨外植体褐变与多酚氧化酶及多酚类物质的关系．四川农业大学学报，1998，16(3)；310～313
原永兵，刘成连等．苹果果皮红色形成的机制．园艺学年评，科学出版社，1995，1：181～218
埃罗．斯耶斯特勒(苏兰)．木材化学．中国林业出版社，1985
徐萌，山仑．旱地作物生产中化学调控技术的研究和应用．山西农业科学，1991，1：31～33
唐章林，张学昆，朱利泉等．甘蓝型油菜雄性不育的酶活性和同工酶谱分析．西南农业大学学报，1999，21(4)：311～316
梁艳丽、梁颖等．甘蓝型黄籽与黑籽种皮多酚及PPO的比较。西南农业大学学报，2002，24(2)：128～130
梁颖等。油菜光合生理指标与产量的关联分析．西南农业大学学报，1999，21(3)：244～247
梁颖等。寡日照下甘蓝型油菜某些光合生理特性的初探．西南农业大学学报，2000，22(1)：24～26
符云鹏，宫长荣，陈秋芳等．抑制剂对白肋烟叶片多酚氧化酶活性的影响．河南农业科学，1997，9：7～8
舒群芳，赵路，李文彬等．植物蛋白电泳分析的方法学研究及技术改进．植物学通报，1998，15(6)：73～78
程必芳，陈宝元．甘蓝型、白菜型油菜和白花芥蓝的同工酶比较研究．中国油料，1991，2：24～27
葛明裕等．木材加工化学．东北林业大学出版社，1985，67～84
董遵，刘敬阳．我国甘蓝型黄籽油菜育种研究进展．华中农业大学学报，1999，18(6)：533～535
薛应龙，欧阳光察，澳绍根．植物苯丙氨酸解氨酶的研究．植物生理学报，1983，9(3)：301～306
鞠志国，原永兵，刘成连等．苹果果皮中酚类物质合成规律的研究．莱阳农学院学报，1992，9：222～225
X.H.波钦诺克(苏)．植物生物化学分析方法．科学出版社，1983，165～166，178～180
A.V.Deynze等．用加倍单体群体研究甘蓝型油菜种子颜色和春化作用的遗传．国外农学——油料作物，1995，1：1～4，8
Arakawa O.Photoregulation of anthocyanin synthesis in applw fruit under UV-B and red light.Plant Cell Physiol,1988,29:1385～1389
Bensadoun A,Weinstein D. Assay of protein in the presence of interfering aterials.Anal Biochem, 1976,70:241-250
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding-Analytical Biochemistry,1976,72(l):248-254
Gtinet A.Rakow G. Repression of seedcoat pigmentation in Ethjopian mustard Ganandian-Journan-of Plant-Science,1997,77(4) :5
Jurgen E,Barbel SH,Knobloch KH,et al.Coordinated changes in enzyme activities of phenyl-propanoid metabolism during the growth of Soybean cell suspension cultures.Biochemica et Biophysica Acta, 1974,362(3) :417-424
Knobloch KH,Hahlbrock K. Isoenzymes of p-coumarate:CoA ligase from cell suspension cultures of Glycin max. Eur J Biochem, 1975,52(2) :311-320
Koukol J.Conn EE. The metabolism of aromatic and properties of the phenylalanine deaminase of Hordeum Vulgare. J Biol Chem,1961,236(10) :2692-2698
Lee,C.Y., Kagan V., Jaworski A.W.,and Brown S.K. Enzymatic browning in relation to phenolic compounds and polyphenoloxidas activity among various peach cultivars. J.Agric.Food Chem., 1990,38:99-101
Lichtenthaler H.K.et al. Determinations of total caroten oids and chlorophylls a and b of leaf extracts in different solvents.Biochem Soc Transac, 1983,11(5) : 1103 ~ 1107
M.Y.coseteng and C.Y.Lee. Change in apple polyphenoloxidase and polyphenol concentr-ations in relation to degree of browning.Journal of Food Science, 1987,52(4) :985~989
Rubery PH,Northcote DH. Site of phenylalanine ammonialyase activity and synthesis of lignin during xylem differentiation.Nature, 1968,219(5160) : 1230-1234
Saure MC. External control of anthocyanin formation.Scientia Hort, 1990,42:181-218
Siegelman HW,Hendricks SB.Photocontrol of anthocyanin synthesis in apple skin.Plant Phy-siol, 1958,33:185-190
Stafford HA. Differences between the lignin-like polymers produced formed by peroxidation of engenol and ferulic acid in leaf section of Phleum.Phleum.Plant Physiol.1960,35(1) :108~114
Stringam.G.R.,et al. Chemical and morphological characteristics associated with seed coat color in rapeseed,Proc.4th.Int.
Theander O. Carbohydrates,polyphends,and lignin in seed hulls of different colors
from turnip rapeseed.J Agri Food Chem,1977,25(2) :270~273
Tibor Fuleki et al. Quantitative methods for anthocyanins.J Food Sci,1968,33(1) :295~298
Wang Hanzhong,Liu Houli. Quantitative variation of antholyanidins, polyphenols, transcinnamic acid and PAL activity in seed hulls of black~and yellow-seeded B.napus.华中农业大学学报1996, 15 (6) : 509-511
Wiermann R. Secondary plant products and cell and tissue differentiation.In:Conn EE.The Bio-chemistry of Plant.New York: Academic Press,1981,7:86~112
Wong T.C.,Luh B.,and Whitaker J.R., Isolation and characterization of polyphenol oxidase isoenzymes of clingstone Pheach.Plant Physiol,1971,48:19~23
Woods,D.L. The association of yellow-seed color with other characters in mustard (Brassica juncea),Cruciferae Newsletter, 1980,5:23 ~24
Yamada Y,Kuboi T,Sato F. Cell differentiation proceedings of symposium on plant tissue culture.Peking,1978,371~379
王三根主编．植物生理研究技术．西南农业大学．2000
王汉中，刘后利．油菜皮壳中多酚氧化酶活性的组织化学鉴定及其与粒色的关系．中国油料，1991，1：30～32
王汉中，刘后利．甘蓝型油菜粒色与种子有关性状间的关系研究．中国油料，1989，11(2)：32～35
王保通，商鸿生．小麦高温抗条锈性表达与木质素合成的关系植物保护学报，1996，23(3)：229～233
王敬文，薛应龙．苯丙氨酸解氨酶在抗马铃薯晚疫病中的作用．植物生理学报，1982，8(1)：35～43
王岩，刘学惠，尹佩玉等．几种天然黑色素分子结构的红外光谱表征研究．分析试验室，1996，15(6)：63～65
王新平，李加新．黑糯米中黑色素提取方法的研究．食品科学，1997，18(2)：44～46
田国忠，李怀方，裘维藩．植物过氧化物酶研究进展．武汉植物学研究，2001，19(4)：332～344
丛斌，杨茂成，粟波等．小麦根尖细胞分化过程中木质素合成及其相关酶的活性变化．复旦学报(自然科学版)，1997，36(5)：550～554
白宝璋，汤学军主编．植物生理学测试技术．中国科学技术出版社，1993
仲飞．红星苹果多酚氧化酶某些特性及其抑制剂的研究．园艺学报，1998，25(2)：184～186
朱广廉等．植物生理学试验．北京大学出版社，1990
朱利泉主编．基础生物化学试验原理与方法．成都科技大学出版社，1993
池方，李树人，田红星．红光对套袋苹果花青素含量的影响．河南农业大学学报，1997，31(2)：174～177
刘后利主编．实用油菜栽培学．上海科学出版社．1987，559～561
李加纳，张学昆，谌利等．不同遗传背景的甘蓝型黄籽油菜粒色遗传初步研究．中国油料作物学报，1998，20(4)：16～19
李崇辉，陈文艺．甘蓝型油菜黄籽分布与种皮颜色变化研究．西南农业大学学报．1998，20(3)：256～259
胡能书等．同工酶技术及应用．湖南科学技术出版社，1985
胡晓君．不同类型油菜品种种皮色素含量的关系的研究(硕士论文)．华中农业大学，1988
张小军，牟方生，黄观武等．棉花生化育种技术的初步研究．中国棉花，1996，23(8)：7～8
张健军，李祥，侯明生．小麦品种对梭条斑花叶病毒病抗病特性的研究．Ⅱ抗病性与植株内多酚氧化酶活性及同工酶酶谱的关系．华中农业大学学报，1996，15(5)：420～425
陈玉萍，高永同，刘后利．甘蓝型黄籽油菜粒色及主要品质性状的动态研究．中国油料，1987，4：10～14
陈玉萍，刘后利．甘蓝型油菜种子油菜种子发育进程中种皮色素含量动态．中国油料，1994，16(4)：13～16
陈玉萍，刘后利．甘蓝型油菜种子油菜种子发育进程中种皮颜色变化．中国油料，1995，17(2)：1～3
陈秀芳，王坤范．桃果实发育中褐变因子变化规律的研究．园艺学报，1995，22(3)：230～234
陈炜，刘维营，雷清等．三粒小麦和普通小麦苗期过氧化物酶同工酶和蛋白质的比较研究．作物学报，1999，25(5)：650～653
陈健中，盛炳成，刘克均．苯丙酸代谢与苹果对轮纹病抗性的关系．果树科学，1996，14(3)：149～152
吴江生，石淑稳，刘后利．甘蓝型油菜黄籽突变体的遗传研究．中国油料作物学报，1998，20(3)：5～8
吴振先，苏美霞，陈维信．贮藏荔枝果皮多酚氧化酶及过氧化物酶与褐变的研究．华南农业大学学报，1998，19(1)：12～15
吴耕西，周宏伟，王健民．鸭梨酶促褐变的生化机制及底物鉴定．园艺学报，1992，19(3)：198～202
余沛涛，薛应龙．植物苯丙氨酸解氨酶的研究．植物生理学报，1983，9(3)：301～306
周旭章，魏开华，陈朝辉等．从黑芝麻中提取黑色素的研究．林产化学通讯，1997，4：17～19
祝军，生吉萍，申林等．苹果花青素形成与PAL活性及蛋白质含量的关系．中国农业大学学报，1997，2(3)：97～99
姚金宝，许才康．甘蓝型黄籽油菜种子油菜种子发育进程中若干特性的研究．中国油料，1995，17(3)：4～6
姜慧芳，孙大容，段乃雄等．利用酯酶同工酶分析花生属种间亲缘关系．中国油料，1989，4：21～23
袁艺．侧耳(平菇)的酯酶、多酚氧化酶及过氧化物酶同工酶的电泳分析．安徽农业大学学报，1997，24(1)：88～92
宴本菊，李焕秀．梨外植体褐变与多酚氧化酶及多酚类物质的关系．四川农业大学学报，1998，16(3)；310～313
原永兵，刘成连等．苹果果皮红色形成的机制．园艺学年评，科学出版社，1995，1：181～218
埃罗．斯耶斯特勒(苏兰)．木材化学．中国林业出版社，1985
徐萌，山仑．旱地作物生产中化学调控技术的研究和应用．山西农业科学，1991，1：31～33
唐章林，张学昆，朱利泉等．甘蓝型油菜雄性不育的酶活性和同工酶谱分析．西南农业大学学报，1999，21(4)：311～316
梁艳丽、梁颖等．甘蓝型黄籽与黑籽种皮多酚及PPO的比较。西南农业大学学报，2002，24(2)：128～130
梁颖等。油菜光合生理指标与产量的关联分析．西南农业大学学报，1999，21(3)：244～247
梁颖等。寡日照下甘蓝型油菜某些光合生理特性的初探．西南农业大学学报，2000，22(1)：24～26
符云鹏，宫长荣，陈秋芳等．抑制剂对白肋烟叶片多酚氧化酶活性的影响．河南农业科学，1997，9：7～8
舒群芳，赵路，李文彬等．植物蛋白电泳分析的方法学研究及技术改进．植物学通报，1998，15(6)：73～78
程必芳，陈宝元．甘蓝型、白菜型油菜和白花芥蓝的同工酶比较研究．中国油料，1991，2：24～27
葛明裕等．木材加工化学．东北林业大学出版社，1985，67～84
董遵，刘敬阳．我国甘蓝型黄籽油菜育种研究进展．华中农业大学学报，1999，18(6)：533～535
薛应龙，欧阳光察，澳绍根．植物苯丙氨酸解氨酶的研究．植物生理学报，1983，9(3)：301～306
鞠志国，原永兵，刘成连等．苹果果皮中酚类物质合成规律的研究．莱阳农学院学报，1992，9：222～225
X.H.波钦诺克(苏)．植物生物化学分析方法．科学出版社，1983，165～166，178～180
A.V.Deynze等．用加倍单体群体研究甘蓝型油菜种子颜色和春化作用的遗传．国外农学——油料作物，1995，1：1～4，8
Arakawa O.Photoregulation of anthocyanin synthesis in applw fruit under UV-B and red light.Plant Cell Physiol,1988,29:1385～1389
Bensadoun A,Weinstein D. Assay of protein in the presence of interfering aterials.Anal Biochem, 1976,70:241-250
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding-Analytical Biochemistry,1976,72(l):248-254
Gtinet A.Rakow G. Repression of seedcoat pigmentation in Ethjopian mustard Ganandian-Journan-of Plant-Science,1997,77(4) :5
Jurgen E,Barbel SH,Knobloch KH,et al.Coordinated changes in enzyme activities of phenyl-propanoid metabolism during the growth of Soybean cell suspension cultures.Biochemica et Biophysica Acta, 1974,362(3) :417-424
Knobloch KH,Hahlbrock K. Isoenzymes of p-coumarate:CoA ligase from cell suspension cultures of Glycin max. Eur J Biochem, 1975,52(2) :311-320
Koukol J.Conn EE. The metabolism of aromatic and properties of the phenylalanine deaminase of Hordeum Vulgare. J Biol Chem,1961,236(10) :2692-2698
Lee,C.Y., Kagan V., Jaworski A.W.,and Brown S.K. Enzymatic browning in relation to phenolic compounds and polyphenoloxidas activity among various peach cultivars. J.Agric.Food Chem., 1990,38:99-101
Lichtenthaler H.K.et al. Determinations of total caroten oids and chlorophylls a and b of leaf extracts in different solvents.Biochem Soc Transac, 1983,11(5) : 1103 ~ 1107
M.Y.coseteng and C.Y.Lee. Change in apple polyphenoloxidase and polyphenol concentr-ations in relation to degree of browning.Journal of Food Science, 1987,52(4) :985~989
Rubery PH,Northcote DH. Site of phenylalanine ammonialyase activity and synthesis of lignin during xylem differentiation.Nature, 1968,219(5160) : 1230-1234
Saure MC. External control of anthocyanin formation.Scientia Hort, 1990,42:181-218
Siegelman HW,Hendricks SB.Photocontrol of anthocyanin synthesis in apple skin.Plant Phy-siol, 1958,33:185-190
Stafford HA. Differences between the lignin-like polymers produced formed by peroxidation of engenol and ferulic acid in leaf section of Phleum.Phleum.Plant Physiol.1960,35(1) :108~114
Stringam.G.R.,et al. Chemical and morphological characteristics associated with seed coat color in rapeseed,Proc.4th.Int.
Theander O. Carbohydrates,polyphends,and lignin in seed hulls of different colors
from turnip rapeseed.J Agri Food Chem,1977,25(2) :270~273
Tibor Fuleki et al. Quantitative methods for anthocyanins.J Food Sci,1968,33(1) :295~298
Wang Hanzhong,Liu Houli. Quantitative variation of antholyanidins, polyphenols, transcinnamic acid and PAL activity in seed hulls of black~and yellow-seeded B.napus.华中农业大学学报1996, 15 (6) : 509-511
Wiermann R. Secondary plant products and cell and tissue differentiation.In:Conn EE.The Bio-chemistry of Plant.New York: Academic Press,1981,7:86~112
Wong T.C.,Luh B.,and Whitaker J.R., Isolation and characterization of polyphenol oxidase isoenzymes of clingstone Pheach.Plant Physiol,1971,48:19~23
Woods,D.L. The association of yellow-seed color with other characters in mustard (Brassica juncea),Cruciferae Newsletter, 1980,5:23 ~24
Yamada Y,Kuboi T,Sato F. Cell differentiation proceedings of symposium on plant tissue culture.Peking,1978,371~379