显性核不育亚麻可育、不育花蕾mRNA差异表达研究及差异片段分析
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摘要
本文利用mRNA差异显示技术,以显性核不育亚麻兄妹交分离出的遗传背景极其相似的可育株和不育株为材料,对可育、不育花蕾中基因差异性表达进行研究,并对差异片段进行反向Northern杂交验证、克隆、测序以及序列分析。主要研究结果如下:
(1)建立了适合核不育亚麻的DDRT-PCR体系。
(2)共获得差异片段52个。其中36个为不育差异片段,其余的16个为可育差异片段。
(3)对这52个差异片段进行反向Northern杂交验证,最终获得了5个不育花蕾阳性片段:G+E07+100bp、G+E07+330bp、G+E07+830bp、G+S273+500bp、A+S267+300bp;2个可育花蕾阳性片段:G+S274-250bp、G+S274-450bp。
(4)这些阳性片段经克隆、测序和BLAST序列分析,结果显示:G+E07+100bp与植物GTP结合蛋白高度同源,在细胞信号传导过程中有调节作用,推测可能与核不育亚麻育性有密切关系;G+E07+330bp为亚麻花蕾的一段cDNA,可能与核不育亚麻花粉发育有关;G+E07+830bp与β-木糖苷酶同源性很高,它可能影响了亚麻花药、花粉中碳水化合物的代谢,从而导致了花粉的败育;G+S273+500bp与NAD+ADP核糖转移酶有很高的同源性;A+S267+300bp与分泌蛋白高度同源,推测它可能参与了核不育亚麻信号传导途径、形态发生、细胞凋亡等过程,最终影响了雄性不育的发生;G+S274-250bp和G+S274-450bp与已知植物的碱基序列和氨基酸序列同源性很低,推测它们可能是与显性核不育亚麻育性有关的新基因。
The mRNA differential expression was compared by mRNA differential display PCR (DDRT-PCR) between male sterile and fertile flower buds of flax. The positive differential fragments were confirmed by Reverse Northern Dot Blotting, and then sequence analysis of these fragments was carried out with the main results as follows:
(1) The optimum conditions of DDRT-PCR system of flax were established.
(2) Among the 52 fragments that had been obtained, 36 fragments were from male sterile flower buds; the other 16 fragments were from male fertile flower buds.
(3) 9 fragments were confirmed by Reverse Northern Dot Blotting. 5 fragments were specific to male sterile flower buds: G+E07+100bp, G+E07+330bp, G+E07+830bp, G+S273 +500bp, A+S267+300bp; 2 other fragments were specific to male fertile flower buds: G+S274-250bp, G+S274-450bp.
(4) The positive fragments were sequenced and analyzed with BLAST. The G+E07+100bp which is homologous to GTP-binding protein of plant suggested that it may play an important role in signal transduction. The G+E07+330bp may be closely related to fertility. The G+E07+830bp, which is most similar to the sequence of Beta-D-xylosidase, may influence carbon hydrate metabolism in flax anther and pollen, resulting in pollen abortion. The G+S273+500bp is homologous to NAD+ADP ribosyltransferase. The A+S267+300bp, homologous to secreted protein, may play roles in signal transduction, Morphogenesis and Apoptosis in flax and finally lead to male sterility. The G+S274-250bp and G+S274-450bp had no significant similarity with any known genes in plants, so they may be newly found genes related to fertility in flax.
(1)建立了适合核不育亚麻的DDRT-PCR体系。
(2)共获得差异片段52个。其中36个为不育差异片段,其余的16个为可育差异片段。
(3)对这52个差异片段进行反向Northern杂交验证,最终获得了5个不育花蕾阳性片段:G+E07+100bp、G+E07+330bp、G+E07+830bp、G+S273+500bp、A+S267+300bp;2个可育花蕾阳性片段:G+S274-250bp、G+S274-450bp。
(4)这些阳性片段经克隆、测序和BLAST序列分析,结果显示:G+E07+100bp与植物GTP结合蛋白高度同源,在细胞信号传导过程中有调节作用,推测可能与核不育亚麻育性有密切关系;G+E07+330bp为亚麻花蕾的一段cDNA,可能与核不育亚麻花粉发育有关;G+E07+830bp与β-木糖苷酶同源性很高,它可能影响了亚麻花药、花粉中碳水化合物的代谢,从而导致了花粉的败育;G+S273+500bp与NAD+ADP核糖转移酶有很高的同源性;A+S267+300bp与分泌蛋白高度同源,推测它可能参与了核不育亚麻信号传导途径、形态发生、细胞凋亡等过程,最终影响了雄性不育的发生;G+S274-250bp和G+S274-450bp与已知植物的碱基序列和氨基酸序列同源性很低,推测它们可能是与显性核不育亚麻育性有关的新基因。
The mRNA differential expression was compared by mRNA differential display PCR (DDRT-PCR) between male sterile and fertile flower buds of flax. The positive differential fragments were confirmed by Reverse Northern Dot Blotting, and then sequence analysis of these fragments was carried out with the main results as follows:
(1) The optimum conditions of DDRT-PCR system of flax were established.
(2) Among the 52 fragments that had been obtained, 36 fragments were from male sterile flower buds; the other 16 fragments were from male fertile flower buds.
(3) 9 fragments were confirmed by Reverse Northern Dot Blotting. 5 fragments were specific to male sterile flower buds: G+E07+100bp, G+E07+330bp, G+E07+830bp, G+S273 +500bp, A+S267+300bp; 2 other fragments were specific to male fertile flower buds: G+S274-250bp, G+S274-450bp.
(4) The positive fragments were sequenced and analyzed with BLAST. The G+E07+100bp which is homologous to GTP-binding protein of plant suggested that it may play an important role in signal transduction. The G+E07+330bp may be closely related to fertility. The G+E07+830bp, which is most similar to the sequence of Beta-D-xylosidase, may influence carbon hydrate metabolism in flax anther and pollen, resulting in pollen abortion. The G+S273+500bp is homologous to NAD+ADP ribosyltransferase. The A+S267+300bp, homologous to secreted protein, may play roles in signal transduction, Morphogenesis and Apoptosis in flax and finally lead to male sterility. The G+S274-250bp and G+S274-450bp had no significant similarity with any known genes in plants, so they may be newly found genes related to fertility in flax.
引文
1 Kaul MLH.Male Sterility in higher plants,Springer-Verla Press,Berlin,1988
2 Sear E R.Genetics and farming[M].USDA Yearbook,1947,245-255.
3 傅廷栋,杨小牛,杨光圣.甘蓝型油菜波里马雄性不育的选育研究[J].华中农业大学学报,1989,(8)3:201-207
4 杨世周.辣椒雄性不育两用系的选育[J].园艺学报,1981,8(3):49-53
5 方智远,孙培田.新甘蓝一代杂种“双金”、“园春”、“晚丰”[J].农业科技通讯,1979,(12):14-15
6 Rhoades M.Cytoplasmic inheritance of male sterility in Zea mays[J].Science, 1930,(73):340-341
7 Jones A.and Emsweller SL.A male-sterile onion[J].Proc Amer Soc Hort Sci, 1936,(34):582-585
8 彭谦,李汝松,吕英华,等.菜心雄性不育研究初报[J].中国蔬菜,1989,(1):22-24
9 柯桂兰,赵稚雅,宋胭脂,等.大白菜异源胞质雄性不育系 CMS3411-7 的选育及应用[J].园艺学报,1992,19(4):333-340
10 李殿荣.甘蓝型油菜雄性不育系、保持系、恢复系选育成功并已大面积推广[J].中国农业科学,1986,19(5):94
11 黄晋玲,安泽伟,樊冬丽.棉花晋A细胞质雄性不育系及其保持系的同工酶分析[J].山西农业大学学报,2001,21(3):233-235
12 Ahokas H. A simple and rapid screening method for the determination of protein and tryptophan in kernel halves and small samples of barley meal[J].J Sci Food Agric,1978,(29):47-52
13 张改生 ,杨天章 .偏 型、粘型 和易型小 麦雄性不 育系的初 步研究 [J]. 作物 学报, 1989,15(1):1-10
14 李传友,伏健民,金德敏,等.小麦胞质突变型雄性不育系85EA与其保持系线粒体DNA的比较研究[J].遗传学报,1999,26(5):558-562
15 Pochard E.Recherches sur lepiment in Rapport de activete.Station de Amelioration des Planter Maraicheres.Montfavet,1985,49-66
16 Shimotsuma M. Cytogenetical studies in the genus Citrullus.VII.Inheritance of several characters in watermelons[J].Breeding,1968,(13):235-240
17 Salgare S A and Suwarna Gawde.Effect of copper sulphate on pollen germination of successive flowers of Vigna unguiculata(L.)[J].Ad.Plant Sci,1989,(2):72-78
18 凌定厚,马镇荣,陈梅芳,等.籼稻体细胞无性系雄性不育突变类型[J].遗传学报, 1991,18(2):132-139
19 Mariani C,Gossele V,de Bcuckeleer M,et al.A chimaeric ribonuclease inhibitor generestores to male sterile plants[J].Nature,1992,357:384-387
20 Van der Meer,Stam M E,van Tunen A J,et al.Antisense Inhibition of Flavonoid Biosynthesis in Petunia Anthers Results in Male Sterility[J].Plant Cell, 1992,(4):253-262
21 曹墨菊,荣廷昭,胡长远.三个新选玉米雄性不育胞质花药败育的超微结构观察[J].四川农业大学学报,2002,20(20):84-87
22 Levings C.S, Pring D.R. Restriction endonuclease analysis of mitochondrial DNA from normal and Texas cytoplasmic male steril emaize [J].Science,1976,193:158-160
23 Scbnable P.S,Wise R.P. The molecular basis of cytoplasmic male sterility and fertility restoration[J].Trends Plant Sci,1999,(3):175-180
24 张海艳,于元杰,周玉亮.植物细胞质雄性不育的生物学研究进展[J].山东农业大学学报(自然科学版),2004,35(1):137-141
25 Graves P.V,Bu D,Velours J,et al.Direct protein sequencing of wheat mitochondrial ATP-synthase submit 9 confirms RNA editing in plants[J].Journal of Molecular Biology,1990,214:1-6
26 Frankel R,Scoewcroft W.R.,Whitfeld P.R.Chloroplast DNA variation of isonuclear male-sterile lines of nicotiana[J].Mol Gen Genet,1979,169:128-135
27 Kadowaki K,Suzuki T,Kazama S.A chimeric gene containing the 5portion of atp6 is associated with cytoplasmic male sterility of rice[J].Mol Gen Genet,1990,224:10-16
28 Liu D.F.The theoretical studies on the restoration for dominant genic male sterility in plants[J].Heriditas,1992,14(6):31-36
29 Aarts M,Dirkse W G,Stiekema W J.Transposon tagging of a male sterility gene in Arabidopsis.Nature,1993,363:715-717
30 Wilson Z A,Morroll S M,Dawson J,et al.The Arabidopsis MALE STERILITY1(MS1) gene is a transcriptional regulator of male gametogenesis.Plant Journal,2001,28:27-39
31 Lakshmi P.M.,Thangaraj M.Effect of growth regulators and chemicals on pollen sterility in TGMS lines of rice[J].Plant Growth Regulation,2005,46(2):117-124
32 Sarria R,Lyznik A,Vallejos E,et al.A cytoplasmic male sterility-associated mitochondrial peptide in common bean is post-translationally regulated[J].Plant Cell,1998,10(7):1217-1228
33 耿二省,土志源,蒋健咸.辣椒雄性不育系小孢子发生的细胞学观[J].园艺学报,1994, 21(2):165-169
34 秦泰辰,刘大文.玉米雄性不育系小孢子发生过程中的细胞学特性[J].上海农业学报,1990,6(1):9-14
35 Mariani C.Induction of male sterility in plants by a chimaeric ribonucleasegene[J].Nature,1990,347:737-741
36 粟翼玫,赵双宜,张燕君,等.萝卜雄性不育系小孢子发生的细胞形态学研究[J].园艺学报,1995,22(4):348-352
37 张宪省,吴淑云,刘连航,等.非菜可育与不育花药和花粉发育的细胞学观察[J].山东农业大学学报,1991,22(3):237-243
38 Liang P,Paradee A B.Differential display of eukaryotic messrnger RNA by means of the polymerase chain reaction[J].Science,1992,257(5072):967-970
39 Liang P,Lidia A,Arthur B P.Distribution and cloning of eukaryotic mRNA by means of differential display:refinements and optimization[J].Nucleic Acids Research,1993,21(14):3269-3275
40 Liang P,Weimin Zhu,Xiaoying Zhang,et al.Differential display using one-base anchored oligo-dT primers[J].Nucleic Acids Research,1994,22(25):5763-5764
41 Bertioli D J,Schlichter U H A,Adams M J,et al.An analysis of differential display shows a strong bias towards high copy number mRNAs[J].Nucleic Acids Research,1995,23(21):4520-4523
42 Sompayrac L,Jane S,Burn T C,et al.Overcoming limitations of the mRNA differential display technique[J].Nucleic Acids Research,1995,23(22):4738-4739
43 张玉强,梅长林.探寻 mRNA 的新方法-差别显示法[J].生物技术,1997,7(1):38-39
44 王学德,朱英国.水稻雄性不育与可育花粉的 mRNA 差异显示和 cDNA 差别片段的分析[J].中国科学 C 辑,1998,28(3):257-263
45 石锐,郭长虹.聚丙烯酰胺凝胶中 DNA 的银染方法[J].生物技术,1998,8(5):46-48
46 Guimaraces M J,Lee F,Zlotnik A,et al.Differential display by PCR:novel findings and applications [J].Nucleic Acids Research,1995,23(10):1832-1833
47 Frohman,M.A.,Dush,M.K.and Martin,G.R.Rapid production of full - length cDNAs from rare transcripts:amplification using a single gene-specific oligonucleotide primer[J].Proc.Natl.Acad.Sci.USA,1988,85:8998-9002
48 Sokolov P,Darwin J P.A rapid and simple PCR-based method for isolation of cDNAs from differentially expressed genes[J].Nucleic Acids Research,1994,22(19):4009-4015
49 崔凯荣,邢更生.利用mRNA差别显示技术分析枸杞体细胞胚发生早期基因的差别表达[J].遗传学报,1998,20(5):16-19
50 庄楚雄,徐是雄,卢永根,等.运用 cDNA 缩减杂交法克隆水稻花粉发育有关的 cDNA[J].科学通报,1999,44(17):1842-1846
51 米志勇,王树生,吴乃虎.水稻低分子量 GTP 结合蛋白基因 osTACD 的分离[J].科学通报,2000,45(19):2047-2055
52 曹家树,叶纨芝,张明,曾广文.白菜核雄性不育两用系花蕾的 mRNA 差别显示及其 cDNA 差异片段分析[J].浙江大学学报(农业与生命科学版),2001,27(6):596-599
53 赵昌平,张立平,李云伏,等.小麦光温敏雄性不育相关基因的DDRT-PCR分析及功能预测[J].中国生物化学与分子生物学报,2007,23(1):56-62
54 孙万仓,党占海,安惠贤等.世界油料作物,兰州大学出版,1991
55 王玉富.我国亚麻生物技术的发展[J].中国麻业,2005,27(2):60-65
56 Bateson W,Cairdyner A.E.Male Sterility in flax[J].Gnet,1921,(11):269-275
57 Gairdyner A E.Male sterility in flax Ⅱ .A case of reciprocal crosses in F2[J].Genet,1929,21:117-124
58 Kumar S,Singh S P.Inheritance of male sterility in some introduced varieties of linseed (Linum.usitassimum L)[J].Indian.J.Agric.Sci,1970,40:184-191
59 Thompson,E.Cytoplasmic male-sterility flax with open corollas[J].J Hered,1977,68:185-187
60 Dang Z-H,Zhang J-P,She X C.Induction male sterility in linseed by antibiotics treatment[J].Chinese Journal of Oil Crop Sciences,2000,22(1):46-48
61 陈鸿山.核不育油用亚麻研究初报[J].华北农学报,1986,1(1):87-91
62 高凤云,张辉,斯钦巴特尔.亚麻显性雄性核不育基因的 RAPD 标记[J].华北农学报,2007,22(1):129-131
63 RodbellM.The role of hormone receptors and GTP2 regulatory proteins in membrane transduction[J].Nature,1980,284:17-22
64 Hasunuma.K,K.Furukawa,K.KTom ita.K,et al.GTP2 binding proteins in etiolated epicotyls of Pisum sativum A laskaG seedlings[J].Biochem Biophy s R es Commun,1987,148:133-139
65 Clement C,Audran J C.In Anther and Pollen From Biology to Biotechnology[M].Springer Heidelberg,1999,69-90
66 Liang L-Z,Zhuang Z-Y.DNA damage in peripheral mononuclear blood cells of ovarian cancer patients during combining chemotherapy with cyclophospha-mide carboplatin[J].Chinese Pharmacol Toxicol,1999,13(2):152-155
2 Sear E R.Genetics and farming[M].USDA Yearbook,1947,245-255.
3 傅廷栋,杨小牛,杨光圣.甘蓝型油菜波里马雄性不育的选育研究[J].华中农业大学学报,1989,(8)3:201-207
4 杨世周.辣椒雄性不育两用系的选育[J].园艺学报,1981,8(3):49-53
5 方智远,孙培田.新甘蓝一代杂种“双金”、“园春”、“晚丰”[J].农业科技通讯,1979,(12):14-15
6 Rhoades M.Cytoplasmic inheritance of male sterility in Zea mays[J].Science, 1930,(73):340-341
7 Jones A.and Emsweller SL.A male-sterile onion[J].Proc Amer Soc Hort Sci, 1936,(34):582-585
8 彭谦,李汝松,吕英华,等.菜心雄性不育研究初报[J].中国蔬菜,1989,(1):22-24
9 柯桂兰,赵稚雅,宋胭脂,等.大白菜异源胞质雄性不育系 CMS3411-7 的选育及应用[J].园艺学报,1992,19(4):333-340
10 李殿荣.甘蓝型油菜雄性不育系、保持系、恢复系选育成功并已大面积推广[J].中国农业科学,1986,19(5):94
11 黄晋玲,安泽伟,樊冬丽.棉花晋A细胞质雄性不育系及其保持系的同工酶分析[J].山西农业大学学报,2001,21(3):233-235
12 Ahokas H. A simple and rapid screening method for the determination of protein and tryptophan in kernel halves and small samples of barley meal[J].J Sci Food Agric,1978,(29):47-52
13 张改生 ,杨天章 .偏 型、粘型 和易型小 麦雄性不 育系的初 步研究 [J]. 作物 学报, 1989,15(1):1-10
14 李传友,伏健民,金德敏,等.小麦胞质突变型雄性不育系85EA与其保持系线粒体DNA的比较研究[J].遗传学报,1999,26(5):558-562
15 Pochard E.Recherches sur lepiment in Rapport de activete.Station de Amelioration des Planter Maraicheres.Montfavet,1985,49-66
16 Shimotsuma M. Cytogenetical studies in the genus Citrullus.VII.Inheritance of several characters in watermelons[J].Breeding,1968,(13):235-240
17 Salgare S A and Suwarna Gawde.Effect of copper sulphate on pollen germination of successive flowers of Vigna unguiculata(L.)[J].Ad.Plant Sci,1989,(2):72-78
18 凌定厚,马镇荣,陈梅芳,等.籼稻体细胞无性系雄性不育突变类型[J].遗传学报, 1991,18(2):132-139
19 Mariani C,Gossele V,de Bcuckeleer M,et al.A chimaeric ribonuclease inhibitor generestores to male sterile plants[J].Nature,1992,357:384-387
20 Van der Meer,Stam M E,van Tunen A J,et al.Antisense Inhibition of Flavonoid Biosynthesis in Petunia Anthers Results in Male Sterility[J].Plant Cell, 1992,(4):253-262
21 曹墨菊,荣廷昭,胡长远.三个新选玉米雄性不育胞质花药败育的超微结构观察[J].四川农业大学学报,2002,20(20):84-87
22 Levings C.S, Pring D.R. Restriction endonuclease analysis of mitochondrial DNA from normal and Texas cytoplasmic male steril emaize [J].Science,1976,193:158-160
23 Scbnable P.S,Wise R.P. The molecular basis of cytoplasmic male sterility and fertility restoration[J].Trends Plant Sci,1999,(3):175-180
24 张海艳,于元杰,周玉亮.植物细胞质雄性不育的生物学研究进展[J].山东农业大学学报(自然科学版),2004,35(1):137-141
25 Graves P.V,Bu D,Velours J,et al.Direct protein sequencing of wheat mitochondrial ATP-synthase submit 9 confirms RNA editing in plants[J].Journal of Molecular Biology,1990,214:1-6
26 Frankel R,Scoewcroft W.R.,Whitfeld P.R.Chloroplast DNA variation of isonuclear male-sterile lines of nicotiana[J].Mol Gen Genet,1979,169:128-135
27 Kadowaki K,Suzuki T,Kazama S.A chimeric gene containing the 5portion of atp6 is associated with cytoplasmic male sterility of rice[J].Mol Gen Genet,1990,224:10-16
28 Liu D.F.The theoretical studies on the restoration for dominant genic male sterility in plants[J].Heriditas,1992,14(6):31-36
29 Aarts M,Dirkse W G,Stiekema W J.Transposon tagging of a male sterility gene in Arabidopsis.Nature,1993,363:715-717
30 Wilson Z A,Morroll S M,Dawson J,et al.The Arabidopsis MALE STERILITY1(MS1) gene is a transcriptional regulator of male gametogenesis.Plant Journal,2001,28:27-39
31 Lakshmi P.M.,Thangaraj M.Effect of growth regulators and chemicals on pollen sterility in TGMS lines of rice[J].Plant Growth Regulation,2005,46(2):117-124
32 Sarria R,Lyznik A,Vallejos E,et al.A cytoplasmic male sterility-associated mitochondrial peptide in common bean is post-translationally regulated[J].Plant Cell,1998,10(7):1217-1228
33 耿二省,土志源,蒋健咸.辣椒雄性不育系小孢子发生的细胞学观[J].园艺学报,1994, 21(2):165-169
34 秦泰辰,刘大文.玉米雄性不育系小孢子发生过程中的细胞学特性[J].上海农业学报,1990,6(1):9-14
35 Mariani C.Induction of male sterility in plants by a chimaeric ribonucleasegene[J].Nature,1990,347:737-741
36 粟翼玫,赵双宜,张燕君,等.萝卜雄性不育系小孢子发生的细胞形态学研究[J].园艺学报,1995,22(4):348-352
37 张宪省,吴淑云,刘连航,等.非菜可育与不育花药和花粉发育的细胞学观察[J].山东农业大学学报,1991,22(3):237-243
38 Liang P,Paradee A B.Differential display of eukaryotic messrnger RNA by means of the polymerase chain reaction[J].Science,1992,257(5072):967-970
39 Liang P,Lidia A,Arthur B P.Distribution and cloning of eukaryotic mRNA by means of differential display:refinements and optimization[J].Nucleic Acids Research,1993,21(14):3269-3275
40 Liang P,Weimin Zhu,Xiaoying Zhang,et al.Differential display using one-base anchored oligo-dT primers[J].Nucleic Acids Research,1994,22(25):5763-5764
41 Bertioli D J,Schlichter U H A,Adams M J,et al.An analysis of differential display shows a strong bias towards high copy number mRNAs[J].Nucleic Acids Research,1995,23(21):4520-4523
42 Sompayrac L,Jane S,Burn T C,et al.Overcoming limitations of the mRNA differential display technique[J].Nucleic Acids Research,1995,23(22):4738-4739
43 张玉强,梅长林.探寻 mRNA 的新方法-差别显示法[J].生物技术,1997,7(1):38-39
44 王学德,朱英国.水稻雄性不育与可育花粉的 mRNA 差异显示和 cDNA 差别片段的分析[J].中国科学 C 辑,1998,28(3):257-263
45 石锐,郭长虹.聚丙烯酰胺凝胶中 DNA 的银染方法[J].生物技术,1998,8(5):46-48
46 Guimaraces M J,Lee F,Zlotnik A,et al.Differential display by PCR:novel findings and applications [J].Nucleic Acids Research,1995,23(10):1832-1833
47 Frohman,M.A.,Dush,M.K.and Martin,G.R.Rapid production of full - length cDNAs from rare transcripts:amplification using a single gene-specific oligonucleotide primer[J].Proc.Natl.Acad.Sci.USA,1988,85:8998-9002
48 Sokolov P,Darwin J P.A rapid and simple PCR-based method for isolation of cDNAs from differentially expressed genes[J].Nucleic Acids Research,1994,22(19):4009-4015
49 崔凯荣,邢更生.利用mRNA差别显示技术分析枸杞体细胞胚发生早期基因的差别表达[J].遗传学报,1998,20(5):16-19
50 庄楚雄,徐是雄,卢永根,等.运用 cDNA 缩减杂交法克隆水稻花粉发育有关的 cDNA[J].科学通报,1999,44(17):1842-1846
51 米志勇,王树生,吴乃虎.水稻低分子量 GTP 结合蛋白基因 osTACD 的分离[J].科学通报,2000,45(19):2047-2055
52 曹家树,叶纨芝,张明,曾广文.白菜核雄性不育两用系花蕾的 mRNA 差别显示及其 cDNA 差异片段分析[J].浙江大学学报(农业与生命科学版),2001,27(6):596-599
53 赵昌平,张立平,李云伏,等.小麦光温敏雄性不育相关基因的DDRT-PCR分析及功能预测[J].中国生物化学与分子生物学报,2007,23(1):56-62
54 孙万仓,党占海,安惠贤等.世界油料作物,兰州大学出版,1991
55 王玉富.我国亚麻生物技术的发展[J].中国麻业,2005,27(2):60-65
56 Bateson W,Cairdyner A.E.Male Sterility in flax[J].Gnet,1921,(11):269-275
57 Gairdyner A E.Male sterility in flax Ⅱ .A case of reciprocal crosses in F2[J].Genet,1929,21:117-124
58 Kumar S,Singh S P.Inheritance of male sterility in some introduced varieties of linseed (Linum.usitassimum L)[J].Indian.J.Agric.Sci,1970,40:184-191
59 Thompson,E.Cytoplasmic male-sterility flax with open corollas[J].J Hered,1977,68:185-187
60 Dang Z-H,Zhang J-P,She X C.Induction male sterility in linseed by antibiotics treatment[J].Chinese Journal of Oil Crop Sciences,2000,22(1):46-48
61 陈鸿山.核不育油用亚麻研究初报[J].华北农学报,1986,1(1):87-91
62 高凤云,张辉,斯钦巴特尔.亚麻显性雄性核不育基因的 RAPD 标记[J].华北农学报,2007,22(1):129-131
63 RodbellM.The role of hormone receptors and GTP2 regulatory proteins in membrane transduction[J].Nature,1980,284:17-22
64 Hasunuma.K,K.Furukawa,K.KTom ita.K,et al.GTP2 binding proteins in etiolated epicotyls of Pisum sativum A laskaG seedlings[J].Biochem Biophy s R es Commun,1987,148:133-139
65 Clement C,Audran J C.In Anther and Pollen From Biology to Biotechnology[M].Springer Heidelberg,1999,69-90
66 Liang L-Z,Zhuang Z-Y.DNA damage in peripheral mononuclear blood cells of ovarian cancer patients during combining chemotherapy with cyclophospha-mide carboplatin[J].Chinese Pharmacol Toxicol,1999,13(2):152-155