偃麦草属成熟胚再生体系的建立及解剖结构研究
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摘要
本试验以长穗偃麦草、中间偃麦草及其种间杂交种为材料,以成熟胚为外植体,研究了不同培养基、接种方式和低温处理时间对偃麦草愈伤组织诱导的影响,探讨了基因型、激素配比和活性炭浓度与愈伤组织分化间的相互关系。试验结果表明:
1)接种时,成熟胚的放置方式应以胚朝上,盾片接触培养基为宜。与胚朝下的接种方式相比,前一种方法可有效提高出愈率。
2)4℃低温处理种子对愈伤组织的诱导有一定作用,作用大小依品种、处理时间长短而异。长穗偃麦草和中间偃麦草经4℃低温处理5小时后,出愈率可分别提高到95.55﹪和89.98﹪。
3)通过各种培养基的比对研究,筛选出适于长穗偃麦草成熟胚培养的最适培养基,其中用于愈伤组织的诱导培养基为MS+CH(500mg/L)+2,4-D(3.0mg/L)+6-BA(0.1mg/L);继代培养基为MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.5mg/L);分化培养基为MS+CH(500mg/L)+6-BA(4.0mg/L)+NAA (0.5mg/L)。
4)适于中间偃麦草成熟胚培养的相关培养基如下:愈伤组织诱导培养基为MS+CH(500mg/L) + 2,4-D(2.0mg/L) +6-BA(0.1mg/L) ;继代培养基为MS +CH(500mg/L) + 2,4-D(2.0mg/L) +6-BA(0.2mg/L) ;分化培养基为MS +CH(500mg/L)+6-BA(3.0mg/L)+NAA(0.5mg/L)
5)适于杂种成熟胚培养的相关培养基为:愈伤组织诱导培养基MS+CH(500mg/L)+ 2,4-D(1.0mg/L) + 6-BA(0.2mg/L) ;继代培养基MS + CH(500mg/L) +2,4-D(2.0mg/L) + 6-BA(0.2mg/L) ;分化培养基MS + CH(500mg/L) +6-BA(4.0mg/L)+NAA(0.5mg/L)+AC(4g/L)
本试验还对三种偃麦草根部和叶片的解剖结构进行了分析,发现三种偃麦草根与叶的结构基本一致,但表现出各自的结构特点。如杂交种根中通道细胞较多,叶片角质层较厚。
The study in this paper using mature embryos of E.elongata、E.intermedia and Elytrigia hybrid (E.elongata X E.intermedia) as explant , cultivate them in different culture medium,way of inoculate and processing time of 4℃to find how to effect the growth of callus, and to find the correlation between genotypes, hormone ,AC and callus differentiation . The results were as follows:
(1) The best way of inoculate mature embryos was: placed embryo upwards and made the scutellum tounched medium completely. This way was propitious to the frequency of callus induction.
(2) Treating seeds at low temperature 4℃had certain effect on callus induction, the effect depended on varieties and treatment time. The treating for 5days was suitable for E.elongata、E.intermedia, and their callus frequency reached 98.51% and 89.66% respectively.
(3) The feasible medium of E.elongata
The callus inducing medium: MS+CH(500mg/L)+2,4-D(3.0mg/L)+6-BA(0.1 mg/L);
Subculture medium: MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.5mg/L); Differentiation medium: MS+CH(500mg/L)+6-BA(4.0mg/L)+NAA (0.5mg/L).
(4) The feasible medium of E.intermedia
The callus inducing medium: MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.1 mg/L);
Subculture medium: MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.2 mg/L); Differentiation medium: MS+CH(500mg/L)+6-BA(3.0mg/L)+NAA (0.5mg/L).
(5) The feasible medium of Elytrigia hybrid (E.elongata X E.intermedia) The callus inducing medium: MS+CH(500mg/L)+2,4-D(1.0mg/L)+6-BA(0.2 mg/L);
Subculture medium: MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.2mg/L ); Differentiation medium: MS+CH(500mg/L)6-BA(4.0mg/L)+NAA (0.5mg/L) +AC(4g/L).
The other study in this paper is about anatomic structure analysis of roots and leaves of Elytrigia which have nearly same structure but different character. For example, in the roots of Elytrigia hybrid (E.elongata X E.intermedia) there are more passage cells, and in the leaves, the cuticles are thicker.
1)接种时,成熟胚的放置方式应以胚朝上,盾片接触培养基为宜。与胚朝下的接种方式相比,前一种方法可有效提高出愈率。
2)4℃低温处理种子对愈伤组织的诱导有一定作用,作用大小依品种、处理时间长短而异。长穗偃麦草和中间偃麦草经4℃低温处理5小时后,出愈率可分别提高到95.55﹪和89.98﹪。
3)通过各种培养基的比对研究,筛选出适于长穗偃麦草成熟胚培养的最适培养基,其中用于愈伤组织的诱导培养基为MS+CH(500mg/L)+2,4-D(3.0mg/L)+6-BA(0.1mg/L);继代培养基为MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.5mg/L);分化培养基为MS+CH(500mg/L)+6-BA(4.0mg/L)+NAA (0.5mg/L)。
4)适于中间偃麦草成熟胚培养的相关培养基如下:愈伤组织诱导培养基为MS+CH(500mg/L) + 2,4-D(2.0mg/L) +6-BA(0.1mg/L) ;继代培养基为MS +CH(500mg/L) + 2,4-D(2.0mg/L) +6-BA(0.2mg/L) ;分化培养基为MS +CH(500mg/L)+6-BA(3.0mg/L)+NAA(0.5mg/L)
5)适于杂种成熟胚培养的相关培养基为:愈伤组织诱导培养基MS+CH(500mg/L)+ 2,4-D(1.0mg/L) + 6-BA(0.2mg/L) ;继代培养基MS + CH(500mg/L) +2,4-D(2.0mg/L) + 6-BA(0.2mg/L) ;分化培养基MS + CH(500mg/L) +6-BA(4.0mg/L)+NAA(0.5mg/L)+AC(4g/L)
本试验还对三种偃麦草根部和叶片的解剖结构进行了分析,发现三种偃麦草根与叶的结构基本一致,但表现出各自的结构特点。如杂交种根中通道细胞较多,叶片角质层较厚。
The study in this paper using mature embryos of E.elongata、E.intermedia and Elytrigia hybrid (E.elongata X E.intermedia) as explant , cultivate them in different culture medium,way of inoculate and processing time of 4℃to find how to effect the growth of callus, and to find the correlation between genotypes, hormone ,AC and callus differentiation . The results were as follows:
(1) The best way of inoculate mature embryos was: placed embryo upwards and made the scutellum tounched medium completely. This way was propitious to the frequency of callus induction.
(2) Treating seeds at low temperature 4℃had certain effect on callus induction, the effect depended on varieties and treatment time. The treating for 5days was suitable for E.elongata、E.intermedia, and their callus frequency reached 98.51% and 89.66% respectively.
(3) The feasible medium of E.elongata
The callus inducing medium: MS+CH(500mg/L)+2,4-D(3.0mg/L)+6-BA(0.1 mg/L);
Subculture medium: MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.5mg/L); Differentiation medium: MS+CH(500mg/L)+6-BA(4.0mg/L)+NAA (0.5mg/L).
(4) The feasible medium of E.intermedia
The callus inducing medium: MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.1 mg/L);
Subculture medium: MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.2 mg/L); Differentiation medium: MS+CH(500mg/L)+6-BA(3.0mg/L)+NAA (0.5mg/L).
(5) The feasible medium of Elytrigia hybrid (E.elongata X E.intermedia) The callus inducing medium: MS+CH(500mg/L)+2,4-D(1.0mg/L)+6-BA(0.2 mg/L);
Subculture medium: MS+CH(500mg/L)+2,4-D(2.0mg/L)+6-BA(0.2mg/L ); Differentiation medium: MS+CH(500mg/L)6-BA(4.0mg/L)+NAA (0.5mg/L) +AC(4g/L).
The other study in this paper is about anatomic structure analysis of roots and leaves of Elytrigia which have nearly same structure but different character. For example, in the roots of Elytrigia hybrid (E.elongata X E.intermedia) there are more passage cells, and in the leaves, the cuticles are thicker.
引文
1 利容千,王明全. 植物组织培养简明教程[M] .武汉:武汉大学出版社,2004
2 Chen C H,Stenberg N E,Ross J G.Clonal propagation of big bluetem by tissue cultere.Crop Sci.1977,17:847-50
3 Wang Z Y,Hopkins A,Mian R.Forage and turf grass biotechnology. Critical Reviews in Piant Sciences.2001,20(6):573-19
4 Spangenberg G,Kalla R,Lidgett A. Transgenesis and genomics in molecular breeding of forage plants. Proceedings of the 10th Australian Agronomy Conference, Hobart,001
5 Dale P J.Embryoids from cultured immature embryos of Lolium multiflorum.Z.Pflanz-Enphysiol.1980.100:73～77
6 Torello W.A,Symington A G, Rufner R. Callus initiatioin plant regeneration and
7 evidence of somatic embryogenesis in red fesucue. Crop SCI. 1984.24:1037～1040
8 Wang Z Y,Legris G,Nage1 J,Potrykus I,Spangenberg G.Cryopreservation of embryogenic cell suspension in Festuca and Lolium species.Plant Science 1994.(103):93～106
9 陈以峰,等.水稻体细胞培养中胚性细胞出现与 IAA 的关系.植物学报,1998. 40 (5): 474～478
10 Ahn B J,Huang F H,King J W.Plant regeneration through somatic embryogenesis in common bermudagrass tissue culture.Crop Sci.1985(25):1107～1109
11 Vasil V. Vasil I.K. Somatic embryogenesis and plant regeneration from tissue cultures of Pennisetum americanum and P.americamum×P.purpureum hybrid. Am.J.Bot. 1981.68: 864～872
12 Murashige T, Skoog F.A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant.1962.15:473～497
13 Linsmaier E M, Skoog f.Organic growth factor requirement for tobacco tissue culture. Phusiol Plant. 1965(18):100～127
14 Gamborg O L,Murashige T,Thorpe T A,Vasil I K,Plant tissue culture media.In Vitro 1976.12:473～478
15 Zaghmout O M F,Torello W A.Somatic embrygenesis and plant regeneration from suspenesis and plant regeneration fromsuspensis cultures of red fescue[J].Crop Sci.1989,29(3):815～817
16 Wang Z Y, Legris G, Nagel J, Potrykus I, Spangenberg G. Cryopreservation of embryogenic cell suspension in Festuca and Loliurn species. Plant Science. 1994, (103):93～106
17 马忠华,张云芳,徐传祥,等.早熟禾的组织培养和基因枪介导的基因转化体系的初步建立[J].复旦学报,1999,38〔5〕:540～544
18 Inokuma C,Sugiura K,Cho C,Okawara R,Kaneko S.Plant regeneration from protoplasta of Japanese lawngrass.Plant Cell Rep.1996,(15):737～741
19 Kasperbeuer M J,Eizenge G C.Tall fescue doubled haploids via tissue culture and plant regeneration.Crop Sci.1985,(25)1091～1095
20 陈默君.中国饲用植物 [M].北京:中国农业出版社,2000
21 谷安琳.耐盐碱栽培牧草—长穗薄冰草[J].中国草地,2004,26〔2〕:9～9
22 孟林,张国芳.优良饲用坪用水土保持兼用植物—偃麦草 [J].草原与草坪,2003,〔4〕:16～17
23 徐柱主编.中国牧草手册 [M].北京:化学工业出版社,2004
24 张国芳 偃麦草属植物光合特性及其抗旱性鉴定研究 中国农科院硕士论文 2006
25 Cauderon R.Genetic analysis in the genus Agropyron.Hereditas Suppl.1966,2:218～234
26 Cauderon R.The resistance to wheat rusts of Agropyron intermedium and its use in wheat improvement. Proc.4th lnt.Wheat Genet.Symp.,1979.pp401～407
27 张相岐等.小冰草异附加系中天兰冰草染色体的变异[J],遗传学报,1991,18(4):344～351
28 Qin Chen, F.ahmad.Genomic constitution of a partial amphiploid OK 7211542 used as a source of immunity to barley yellow dwarf virus fot bread wheat.Plant Breeding.1998,117.1～6
29 Dewey DR.Geneome analysis of Agropyron repens × Agropyron cri statum synthetic hybrids.Amer.J.Bot.1964,51:1062～1068
30 30 Dewey DR. Synthetic hybrid of new world and old word Agropyron Ⅳ.Inermex tetraploid Agropyron deserlorum.Amer.J. Bot.1967,54:403～409
31 李振声.小麦远缘杂交[M]科学出版社.北京.1985
32 李玉京,刘建中,李兵等.长穗偃麦草基因组中与耐低磷营养胁迫有关的基因染色体定位[J].遗传学报,1999,26〔6〕:703—710
33 易津,谷安琳,贾光宏等.赖草属牧草幼苗耐旱性生理基础的研究[J].干旱区资源与环境,2001,15〔5〕:47～50
34 H.B.齐津(胡启德译).植物远缘杂交[M] .科学出版社.北京.1957
35 孙善澄.小麦与偃麦草远缘杂交的研究[J] .华北农学报,1987,2(1):7～12
36 薛秀庄,王祥正.八倍体小偃麦中 3、中 4、中 5 在小麦育种中的应用[J] .陕西农业科学,1989,2(1):7～12
37 李振声.蓝粒单体小麦研究[J] .遗传学报,1982,9(6):431～439
38 李振声.小麦远缘杂交与染色体工程[M] .科学出版社
39 庄巧生,王恒立主编.小麦育种理论与实践的进展.科学普及出版社.1987
40 武东亮,辛志勇等.抗黄矮病普通小麦 2 偃麦草异附加系、异代换系的选育和鉴定 [J] .中国科学(C 辑),1999,29(1):62～67
41 张荣琦,陈春环,赵晓农等.八倍体小偃麦与普通小麦杂交选育抗旱小麦品种的研究[J] .西北植物学报,1999,19(1):8～24
42 Linacero R,etal.Somatic embryogenesis from immature infortscences of rye[J].Plant Sci.1990,(72):253～258
43 刘晓东,傅体华,任正隆.小麦与八倍体小偃麦杂种结合幅射诱导后代的细胞学分析[J].四川农业大学学报,2005,25〔3〕:15～18
44 庄丽芳,亓增军,英加,陈佩度,刘大均.普通小麦–百萨偃麦草二体异附加系的选育与鉴定[J].遗传学报,2003,30〔10〕:919～925
45 朱诚,吴运荣,金维正,等.胚乳对籼稻成熟胚愈伤组织诱导及分化的影响[J].浙江农业学报,2001,13〔1〕:13～15
46 林拥军,邹玉兰,万骅,等.水稻成熟胚组培过程中胚乳影响的研究[J].江西农业大学学报,1996,18〔3〕:259～263
47 解继红,云锦凤,杨斌,侯建华.2,4-D 和 BAP 对蒙古冰草幼胚愈伤组织诱导及生长的影响[J].中国草地学报,2006,28〔2〕:44～47
48 柳建军,于洪欣,冯兆礼.小麦成熟胚愈伤组织诱导及分化的研究[J].山东大学学报,1996,27(4):451～456]
49 丁莉萍,何光源.小麦成熟胚高频植株再生系统的建立[J].植物生理学通报,2004,40(2):187～190
50 廖祥儒,杜建芳,王俊霞,等.预处理对小麦成熟胚愈伤组织形成的影响[J].河北大学学报,1999,9(1):41～44
51 廖祥儒,郭中伟,杜建芳,等.低温和 PEG 预处理对小麦愈伤组织形成及 IAA 氧化的影响[J].河北大学学报,1999,9(1):41～-44
52 陈军营,尹海燕,梁静静,等.不同预处理对小麦成熟胚愈伤组织形成的影响[J].河北大学学报,2005,(1):19～21
53 钱海丰,等.激素对高羊芋愈伤组织诱导及其分化的影响.中国草地,2002, 24 (1): 46～50
54 Griffin J D,et al. High-frequency plant regeneration from seedderived callus cultures of Kentuckybluegrass (Poa pratensis L.)[J]. Plant Cell Reports, 1995,14: 721～724
55 钱海丰, 薛庆中, 高羊茅的组织培养和植株再生[J]. 植物生理学讯.2002,38(3):247～247
56 金洪, 张众, 云锦凤, 黄素玲, 吴平.诺丹冰草成熟胚离体培养中不定芽的诱导和形态建成[J].中国草地.1998,No.3, pp41～43, 52
57 贺杰,校现周,李瑞芬.结缕草成熟胚愈伤组织的诱导及再生体系的研究[J].山西农业大学学报,2005,25〔3〕:211～213
58 张伟,简玉瑜,杨跃生.栽培稻与野生稻原生质体培养与融合[A].农业科学集刊(第二集刊),农作物原生质培养专辑[C].北京:农业出版社,1995.27～32
59 胡张华,陈火庆,吴关庭,等.百慕大成熟胚组织培养及植株再生[J].草业学报,2003,12〔1〕:85～89
60 王海波,魏景芳,葛亚新,石西平,范云六.小麦愈伤组织状态调控与原生质体培养[J].中国农业科学,1996,29〔6〕:8～14
61 Zhang L J , etal.Efficient transformation of tobacco by ultrasonicatio. Bio/Technology. 1991,9:996～996
62 田文忠.提高粕稻愈伤组织再生频率的研究[J].遗传学报,1994, 21(1): 215～221
63 Linacero R, et al. Somatic embryogenesis from immature inforescences of rye [J].Plant Sci. 1990, 72: 253～258
64 陈以峰,等.水稻体细胞培养中胚性细胞出现于 IAA 的关系[J].植物学报,1998, 40 (5): 474
65 过全生,孙晓丽,韦小敏,胡彦民.低浓度 2, 4-D 提高水稻体细胞成苗研究[J].中国水稻科学,1996, 10(1): 103～109
66 K.伊稍著,李正理译.种子植物解剖结构学(第二版)[M].上海科学出版社,1982,245～261
67 刘穆著.种子植物形态解剖学导论(第二版)[M].科学出版社,2001,247～254
68 Weatherhead M A,Burdon J,Henshaw G G.Some effects of activated charcoal as an additive to piant tissue culturemedia[J].Z pflanzen-physiol,1978 (89): 141～147
69 周智彬,李培军.我国旱生植物的形态解剖学研究[J].干旱区研究,2002, 19(1): 35～40
70 王勋龄,王静.植物形态结构与环境[M].兰州大学出版社,1989,57～69
71 王艳,张绵.结缕草和早熟禾解剖结构与其抗旱性、耐践踏性和弹性的对比研究[J].辽宁大学学报,2000,27(4):372～375
72 王六英,赵金花.偃麦草属三种牧草营养器官解剖结构与抗旱性的研究[J].干旱区资源与环境,2001, 15(5):63～67
73 王怡.三种抗旱植物叶片解剖结构的对比观察[J].四川林业科技,2003,24(1):64～67
74 耿世磊,吴鸿等.三种草坪草的茎、叶解剖结构及其坪用性状[J].热带亚热带植物学报,2002,10(2):145～151
75 王维,吴晓亮,等.小麦抗旱型的形态性状及初生根解剖结构的研究[J].种子,2002,126(6):14～15
2 Chen C H,Stenberg N E,Ross J G.Clonal propagation of big bluetem by tissue cultere.Crop Sci.1977,17:847-50
3 Wang Z Y,Hopkins A,Mian R.Forage and turf grass biotechnology. Critical Reviews in Piant Sciences.2001,20(6):573-19
4 Spangenberg G,Kalla R,Lidgett A. Transgenesis and genomics in molecular breeding of forage plants. Proceedings of the 10th Australian Agronomy Conference, Hobart,001
5 Dale P J.Embryoids from cultured immature embryos of Lolium multiflorum.Z.Pflanz-Enphysiol.1980.100:73～77
6 Torello W.A,Symington A G, Rufner R. Callus initiatioin plant regeneration and
7 evidence of somatic embryogenesis in red fesucue. Crop SCI. 1984.24:1037～1040
8 Wang Z Y,Legris G,Nage1 J,Potrykus I,Spangenberg G.Cryopreservation of embryogenic cell suspension in Festuca and Lolium species.Plant Science 1994.(103):93～106
9 陈以峰,等.水稻体细胞培养中胚性细胞出现与 IAA 的关系.植物学报,1998. 40 (5): 474～478
10 Ahn B J,Huang F H,King J W.Plant regeneration through somatic embryogenesis in common bermudagrass tissue culture.Crop Sci.1985(25):1107～1109
11 Vasil V. Vasil I.K. Somatic embryogenesis and plant regeneration from tissue cultures of Pennisetum americanum and P.americamum×P.purpureum hybrid. Am.J.Bot. 1981.68: 864～872
12 Murashige T, Skoog F.A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant.1962.15:473～497
13 Linsmaier E M, Skoog f.Organic growth factor requirement for tobacco tissue culture. Phusiol Plant. 1965(18):100～127
14 Gamborg O L,Murashige T,Thorpe T A,Vasil I K,Plant tissue culture media.In Vitro 1976.12:473～478
15 Zaghmout O M F,Torello W A.Somatic embrygenesis and plant regeneration from suspenesis and plant regeneration fromsuspensis cultures of red fescue[J].Crop Sci.1989,29(3):815～817
16 Wang Z Y, Legris G, Nagel J, Potrykus I, Spangenberg G. Cryopreservation of embryogenic cell suspension in Festuca and Loliurn species. Plant Science. 1994, (103):93～106
17 马忠华,张云芳,徐传祥,等.早熟禾的组织培养和基因枪介导的基因转化体系的初步建立[J].复旦学报,1999,38〔5〕:540～544
18 Inokuma C,Sugiura K,Cho C,Okawara R,Kaneko S.Plant regeneration from protoplasta of Japanese lawngrass.Plant Cell Rep.1996,(15):737～741
19 Kasperbeuer M J,Eizenge G C.Tall fescue doubled haploids via tissue culture and plant regeneration.Crop Sci.1985,(25)1091～1095
20 陈默君.中国饲用植物 [M].北京:中国农业出版社,2000
21 谷安琳.耐盐碱栽培牧草—长穗薄冰草[J].中国草地,2004,26〔2〕:9～9
22 孟林,张国芳.优良饲用坪用水土保持兼用植物—偃麦草 [J].草原与草坪,2003,〔4〕:16～17
23 徐柱主编.中国牧草手册 [M].北京:化学工业出版社,2004
24 张国芳 偃麦草属植物光合特性及其抗旱性鉴定研究 中国农科院硕士论文 2006
25 Cauderon R.Genetic analysis in the genus Agropyron.Hereditas Suppl.1966,2:218～234
26 Cauderon R.The resistance to wheat rusts of Agropyron intermedium and its use in wheat improvement. Proc.4th lnt.Wheat Genet.Symp.,1979.pp401～407
27 张相岐等.小冰草异附加系中天兰冰草染色体的变异[J],遗传学报,1991,18(4):344～351
28 Qin Chen, F.ahmad.Genomic constitution of a partial amphiploid OK 7211542 used as a source of immunity to barley yellow dwarf virus fot bread wheat.Plant Breeding.1998,117.1～6
29 Dewey DR.Geneome analysis of Agropyron repens × Agropyron cri statum synthetic hybrids.Amer.J.Bot.1964,51:1062～1068
30 30 Dewey DR. Synthetic hybrid of new world and old word Agropyron Ⅳ.Inermex tetraploid Agropyron deserlorum.Amer.J. Bot.1967,54:403～409
31 李振声.小麦远缘杂交[M]科学出版社.北京.1985
32 李玉京,刘建中,李兵等.长穗偃麦草基因组中与耐低磷营养胁迫有关的基因染色体定位[J].遗传学报,1999,26〔6〕:703—710
33 易津,谷安琳,贾光宏等.赖草属牧草幼苗耐旱性生理基础的研究[J].干旱区资源与环境,2001,15〔5〕:47～50
34 H.B.齐津(胡启德译).植物远缘杂交[M] .科学出版社.北京.1957
35 孙善澄.小麦与偃麦草远缘杂交的研究[J] .华北农学报,1987,2(1):7～12
36 薛秀庄,王祥正.八倍体小偃麦中 3、中 4、中 5 在小麦育种中的应用[J] .陕西农业科学,1989,2(1):7～12
37 李振声.蓝粒单体小麦研究[J] .遗传学报,1982,9(6):431～439
38 李振声.小麦远缘杂交与染色体工程[M] .科学出版社
39 庄巧生,王恒立主编.小麦育种理论与实践的进展.科学普及出版社.1987
40 武东亮,辛志勇等.抗黄矮病普通小麦 2 偃麦草异附加系、异代换系的选育和鉴定 [J] .中国科学(C 辑),1999,29(1):62～67
41 张荣琦,陈春环,赵晓农等.八倍体小偃麦与普通小麦杂交选育抗旱小麦品种的研究[J] .西北植物学报,1999,19(1):8～24
42 Linacero R,etal.Somatic embryogenesis from immature infortscences of rye[J].Plant Sci.1990,(72):253～258
43 刘晓东,傅体华,任正隆.小麦与八倍体小偃麦杂种结合幅射诱导后代的细胞学分析[J].四川农业大学学报,2005,25〔3〕:15～18
44 庄丽芳,亓增军,英加,陈佩度,刘大均.普通小麦–百萨偃麦草二体异附加系的选育与鉴定[J].遗传学报,2003,30〔10〕:919～925
45 朱诚,吴运荣,金维正,等.胚乳对籼稻成熟胚愈伤组织诱导及分化的影响[J].浙江农业学报,2001,13〔1〕:13～15
46 林拥军,邹玉兰,万骅,等.水稻成熟胚组培过程中胚乳影响的研究[J].江西农业大学学报,1996,18〔3〕:259～263
47 解继红,云锦凤,杨斌,侯建华.2,4-D 和 BAP 对蒙古冰草幼胚愈伤组织诱导及生长的影响[J].中国草地学报,2006,28〔2〕:44～47
48 柳建军,于洪欣,冯兆礼.小麦成熟胚愈伤组织诱导及分化的研究[J].山东大学学报,1996,27(4):451～456]
49 丁莉萍,何光源.小麦成熟胚高频植株再生系统的建立[J].植物生理学通报,2004,40(2):187～190
50 廖祥儒,杜建芳,王俊霞,等.预处理对小麦成熟胚愈伤组织形成的影响[J].河北大学学报,1999,9(1):41～44
51 廖祥儒,郭中伟,杜建芳,等.低温和 PEG 预处理对小麦愈伤组织形成及 IAA 氧化的影响[J].河北大学学报,1999,9(1):41～-44
52 陈军营,尹海燕,梁静静,等.不同预处理对小麦成熟胚愈伤组织形成的影响[J].河北大学学报,2005,(1):19～21
53 钱海丰,等.激素对高羊芋愈伤组织诱导及其分化的影响.中国草地,2002, 24 (1): 46～50
54 Griffin J D,et al. High-frequency plant regeneration from seedderived callus cultures of Kentuckybluegrass (Poa pratensis L.)[J]. Plant Cell Reports, 1995,14: 721～724
55 钱海丰, 薛庆中, 高羊茅的组织培养和植株再生[J]. 植物生理学讯.2002,38(3):247～247
56 金洪, 张众, 云锦凤, 黄素玲, 吴平.诺丹冰草成熟胚离体培养中不定芽的诱导和形态建成[J].中国草地.1998,No.3, pp41～43, 52
57 贺杰,校现周,李瑞芬.结缕草成熟胚愈伤组织的诱导及再生体系的研究[J].山西农业大学学报,2005,25〔3〕:211～213
58 张伟,简玉瑜,杨跃生.栽培稻与野生稻原生质体培养与融合[A].农业科学集刊(第二集刊),农作物原生质培养专辑[C].北京:农业出版社,1995.27～32
59 胡张华,陈火庆,吴关庭,等.百慕大成熟胚组织培养及植株再生[J].草业学报,2003,12〔1〕:85～89
60 王海波,魏景芳,葛亚新,石西平,范云六.小麦愈伤组织状态调控与原生质体培养[J].中国农业科学,1996,29〔6〕:8～14
61 Zhang L J , etal.Efficient transformation of tobacco by ultrasonicatio. Bio/Technology. 1991,9:996～996
62 田文忠.提高粕稻愈伤组织再生频率的研究[J].遗传学报,1994, 21(1): 215～221
63 Linacero R, et al. Somatic embryogenesis from immature inforescences of rye [J].Plant Sci. 1990, 72: 253～258
64 陈以峰,等.水稻体细胞培养中胚性细胞出现于 IAA 的关系[J].植物学报,1998, 40 (5): 474
65 过全生,孙晓丽,韦小敏,胡彦民.低浓度 2, 4-D 提高水稻体细胞成苗研究[J].中国水稻科学,1996, 10(1): 103～109
66 K.伊稍著,李正理译.种子植物解剖结构学(第二版)[M].上海科学出版社,1982,245～261
67 刘穆著.种子植物形态解剖学导论(第二版)[M].科学出版社,2001,247～254
68 Weatherhead M A,Burdon J,Henshaw G G.Some effects of activated charcoal as an additive to piant tissue culturemedia[J].Z pflanzen-physiol,1978 (89): 141～147
69 周智彬,李培军.我国旱生植物的形态解剖学研究[J].干旱区研究,2002, 19(1): 35～40
70 王勋龄,王静.植物形态结构与环境[M].兰州大学出版社,1989,57～69
71 王艳,张绵.结缕草和早熟禾解剖结构与其抗旱性、耐践踏性和弹性的对比研究[J].辽宁大学学报,2000,27(4):372～375
72 王六英,赵金花.偃麦草属三种牧草营养器官解剖结构与抗旱性的研究[J].干旱区资源与环境,2001, 15(5):63～67
73 王怡.三种抗旱植物叶片解剖结构的对比观察[J].四川林业科技,2003,24(1):64～67
74 耿世磊,吴鸿等.三种草坪草的茎、叶解剖结构及其坪用性状[J].热带亚热带植物学报,2002,10(2):145～151
75 王维,吴晓亮,等.小麦抗旱型的形态性状及初生根解剖结构的研究[J].种子,2002,126(6):14～15