小麦成熟胚离体培养研究
|
摘要
本研究采用四川省重点推广的小麦新品种R57(川农17)和R59(川农12)的成熟胚及R59的去胚芽成熟胚为外植体，进行离体培养研究，以建立高效的小麦再生体系，从而解决小麦组织培养过程中外植体来源受季节限制的问题，并为进一步进行小麦转基因研究打下基础。同时也将两个品种的组织培养特性与Bobwhite的组织培养特性作比较。现将本研究结果概括如下：
1．本研究建立了以小麦成熟胚为外植体的高效适用的小麦植株再生体系，即以完整成熟胚为外植体，在诱导培养基中添加2mg/l 2,4-D，将经过14天诱导的愈伤组织，于附加5mg／lKT的分化培养基中进行分化培养，这样的组合能取得最好的效果。
2．R57、R59两个品种的成熟胚有相似的优良组织培养特性，其成愈率和愈伤组织分化率在同类研究中位居前列。在不同诱导培养基中，R.57成熟胚平均成愈率最低为77.28％，最高达97.58％。R59成熟胚最低平均成愈率为79.22％，最高为99.33％。在最适宜的培养条件下，R57愈伤组织平均分化率为73.63％。R59愈伤组织平均分化率可达89.23％。
3．种子的收获时期和培养前的低温处理对成熟胚愈伤组织的诱导效应有影响。2002年收获的且经低温处理的成熟胚比2001年收获的未经低温处理的成熟胚有更高的成愈率，且在大多数诱导培养基中差异显著。2001年的成熟胚在不同诱导培养基中的成愈率存在显著差异，而2002年的成熟胚在不同诱导培养基中的成愈率差异不显著。
4．本文首次报道了种子的收获时期和培养前的低温处理对愈伤组织诱导过程中成熟胚萌发成苗的影响。2002年的成熟胚比2001的成熟胚有更高的发芽率，且差异极显著。本研究还表明，高浓度2,4-D有抑制成熟胚萌发的作用，KT有促进成熟胚萌发的作用。
5．本文首次报道了胚芽对成熟胚愈伤组织的形成及愈伤组织的分化能力的影响。胚芽对愈伤组织的形成和愈伤组织的分化都有利。R59去胚芽成熟胚的成愈率低于完整成熟胚的成愈率，且在多数诱导培养基中存在显著或极显著差异。Bobwhite去胚芽成熟胚的成愈率和完整成熟胚的成愈率无差异，都达100％。然而，完整成熟
胚来源的愈伤组织比去胚芽成熟胚来源的愈伤组织分化率更高，且两者存在极显著
差异。
6.R57、R59和BobW址加的组织培养特性有一定的差异。在相同培养条件下，
Bob认七i仍成熟胚的成愈率达1 00%，愈伤组织分化率为94.63%，和R57相比，两者
存在显着差异。但和R59相比不存在显著差异。对去胚芽成熟胚而言，Bobwhi忆的
成愈率为100%，分化率为21 .6%，R59的成愈率为91.93%，分化率为n.67%，在
愈伤组织形成能力方面两者不存在显著差异，但Bobw坛妞的愈伤组织分化能力比
R59强，且两者存在显著差异。
7.综上研究结果表明，R57、R59成熟胚不仅具有很高的成愈率和分化率。而
且两个品种的农艺性状优良。因此，这两个小麦新品种是小麦转基因的良好受体。
本研究建立的小麦商效再生体系可以为其他研究者提供参考。
Mature wheat embryos of new wheat varieties R57(ChuanNongl7) and R59(ChuanNongl2) which are important cultivar in Sichuan Province were used as explants for culture in vitro.And the mature embryos of other wheat genotype"Bobwhite" were also used to culture. When mature embryos were inoculated ,some embryonic buds of RS9 and Bobwhite' s mature embryos were removed.The purpose of this study is to construct a high effective system of plantlet regeneration for mature wheat embryos.In this study,the response of RS7 and RS9 to culture was compared with the response of Bobwhite to culture.The general result is following:
In this study,a high effective wheat plantlet regeneration system was built. This experiment showed that the callus induction medium supplemented with 2mg/l 2,4-D,callus induction time of 14 days and regeneration medium with Smg/1 KT were the best combination and the best explants were mature embryos with embryonic buds.
R57 and R59 have a good response to culture.The rate of callus formation of R57' s mature embryos is 77.28%~97.58%,as for R59,the rate is 79.22%-99.33%.The highest frequency of plantlet regeneration of callus initiated from R57' s mature embryos is 73.63%,as for R59,the highest frequency is 89.23%.Few researchers got the same result in previous studies.
Harvesting time and treating by low temperature had effect on callus formation.The rate of callus formation of mature embryos excised from seeds harvested hi 2002 was higher than that of mature embryos excised from seeds harvested in 2001 .There was no significant difference in different callus induction media for callus formation of mature embryos harvested hi 2001.But great difference hi different callus induction media was
observed for callus formation of mature embryos harvested in 2002.
During the callus induction, the germination of mature embryos was affected by harvesting time and low temperature treat. It is the first time to report this kind of result The rate of germination of mature embryos harvested hi 2002 was by far higher than that of mature embryos harvested in 2001. In this study,high density of 2,4-D could prevent mature embryos from germinating,but KT could prompt germination.
Embryonic bud profited callus formation and differentiation.For R59,significant difference between mature embryos with embryonic buds and those without embryonic buds was observed for callus initiation and differentiation.For Bobwhite,there was no difference between the two kinds of mature embryos for callus formation,but great diference was observed for plantlet regeneration.There were no reports about the effect of embryonic bud on callus initiation and plantlet regneration in previous studies.
There was great difference between RS7 and Bobwhite for callus initiation and plantlet regeneration from mature embryos with embryonic bud.Bobwhite was better than R57.But for the same case.there was no great difference between R59 and Bobwhite.So far as mature embryos without embryonic buds was concerned,there was no significant difference between R59 and Bobwhite for callus formation,but great dufference was observed for plantlet regeneration and Bobwhite was better than R59.
The new wheat varieties R57 and R59 have an excellent character for culture in vitro.And they have good agricultural trait.So they are good acceptors for transgenic wheatThe method of mature wheat embryos culture in vitro found in this study can be reference for other researchers.
1．本研究建立了以小麦成熟胚为外植体的高效适用的小麦植株再生体系，即以完整成熟胚为外植体，在诱导培养基中添加2mg/l 2,4-D，将经过14天诱导的愈伤组织，于附加5mg／lKT的分化培养基中进行分化培养，这样的组合能取得最好的效果。
2．R57、R59两个品种的成熟胚有相似的优良组织培养特性，其成愈率和愈伤组织分化率在同类研究中位居前列。在不同诱导培养基中，R.57成熟胚平均成愈率最低为77.28％，最高达97.58％。R59成熟胚最低平均成愈率为79.22％，最高为99.33％。在最适宜的培养条件下，R57愈伤组织平均分化率为73.63％。R59愈伤组织平均分化率可达89.23％。
3．种子的收获时期和培养前的低温处理对成熟胚愈伤组织的诱导效应有影响。2002年收获的且经低温处理的成熟胚比2001年收获的未经低温处理的成熟胚有更高的成愈率，且在大多数诱导培养基中差异显著。2001年的成熟胚在不同诱导培养基中的成愈率存在显著差异，而2002年的成熟胚在不同诱导培养基中的成愈率差异不显著。
4．本文首次报道了种子的收获时期和培养前的低温处理对愈伤组织诱导过程中成熟胚萌发成苗的影响。2002年的成熟胚比2001的成熟胚有更高的发芽率，且差异极显著。本研究还表明，高浓度2,4-D有抑制成熟胚萌发的作用，KT有促进成熟胚萌发的作用。
5．本文首次报道了胚芽对成熟胚愈伤组织的形成及愈伤组织的分化能力的影响。胚芽对愈伤组织的形成和愈伤组织的分化都有利。R59去胚芽成熟胚的成愈率低于完整成熟胚的成愈率，且在多数诱导培养基中存在显著或极显著差异。Bobwhite去胚芽成熟胚的成愈率和完整成熟胚的成愈率无差异，都达100％。然而，完整成熟
胚来源的愈伤组织比去胚芽成熟胚来源的愈伤组织分化率更高，且两者存在极显著
差异。
6.R57、R59和BobW址加的组织培养特性有一定的差异。在相同培养条件下，
Bob认七i仍成熟胚的成愈率达1 00%，愈伤组织分化率为94.63%，和R57相比，两者
存在显着差异。但和R59相比不存在显著差异。对去胚芽成熟胚而言，Bobwhi忆的
成愈率为100%，分化率为21 .6%，R59的成愈率为91.93%，分化率为n.67%，在
愈伤组织形成能力方面两者不存在显著差异，但Bobw坛妞的愈伤组织分化能力比
R59强，且两者存在显著差异。
7.综上研究结果表明，R57、R59成熟胚不仅具有很高的成愈率和分化率。而
且两个品种的农艺性状优良。因此，这两个小麦新品种是小麦转基因的良好受体。
本研究建立的小麦商效再生体系可以为其他研究者提供参考。
Mature wheat embryos of new wheat varieties R57(ChuanNongl7) and R59(ChuanNongl2) which are important cultivar in Sichuan Province were used as explants for culture in vitro.And the mature embryos of other wheat genotype"Bobwhite" were also used to culture. When mature embryos were inoculated ,some embryonic buds of RS9 and Bobwhite' s mature embryos were removed.The purpose of this study is to construct a high effective system of plantlet regeneration for mature wheat embryos.In this study,the response of RS7 and RS9 to culture was compared with the response of Bobwhite to culture.The general result is following:
In this study,a high effective wheat plantlet regeneration system was built. This experiment showed that the callus induction medium supplemented with 2mg/l 2,4-D,callus induction time of 14 days and regeneration medium with Smg/1 KT were the best combination and the best explants were mature embryos with embryonic buds.
R57 and R59 have a good response to culture.The rate of callus formation of R57' s mature embryos is 77.28%~97.58%,as for R59,the rate is 79.22%-99.33%.The highest frequency of plantlet regeneration of callus initiated from R57' s mature embryos is 73.63%,as for R59,the highest frequency is 89.23%.Few researchers got the same result in previous studies.
Harvesting time and treating by low temperature had effect on callus formation.The rate of callus formation of mature embryos excised from seeds harvested hi 2002 was higher than that of mature embryos excised from seeds harvested in 2001 .There was no significant difference in different callus induction media for callus formation of mature embryos harvested hi 2001.But great difference hi different callus induction media was
observed for callus formation of mature embryos harvested in 2002.
During the callus induction, the germination of mature embryos was affected by harvesting time and low temperature treat. It is the first time to report this kind of result The rate of germination of mature embryos harvested hi 2002 was by far higher than that of mature embryos harvested in 2001. In this study,high density of 2,4-D could prevent mature embryos from germinating,but KT could prompt germination.
Embryonic bud profited callus formation and differentiation.For R59,significant difference between mature embryos with embryonic buds and those without embryonic buds was observed for callus initiation and differentiation.For Bobwhite,there was no difference between the two kinds of mature embryos for callus formation,but great diference was observed for plantlet regeneration.There were no reports about the effect of embryonic bud on callus initiation and plantlet regneration in previous studies.
There was great difference between RS7 and Bobwhite for callus initiation and plantlet regeneration from mature embryos with embryonic bud.Bobwhite was better than R57.But for the same case.there was no great difference between R59 and Bobwhite.So far as mature embryos without embryonic buds was concerned,there was no significant difference between R59 and Bobwhite for callus formation,but great dufference was observed for plantlet regeneration and Bobwhite was better than R59.
The new wheat varieties R57 and R59 have an excellent character for culture in vitro.And they have good agricultural trait.So they are good acceptors for transgenic wheatThe method of mature wheat embryos culture in vitro found in this study can be reference for other researchers.
引文
1．安海龙，卫志明等。小麦幼胚培养高效成株系统的建立。植物生理学报，2000，26(6)：532—538。
2．催凯荣，小麦体细胞胚发生中DNA、RNA和蛋白质的合成动态。核农学报，1997，11(4)：209—214。
3．程在全，王芳等。提高小麦体细胞诱导出胚性愈伤组织的方法研究。西南农业学报，1999，12(3)：4—8。
4．陈以峰，汤日圣。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—477。
5．成卓敏，吴茂森等。应用基因枪法获得抗大麦黄矮病毒转基因植株。植物病理学报，2000，30(2)：116—121。
6．关畅，曾寒冰等。小麦未熟胚离体培养的研究。东北农学院学报，1993，24(2)：107—116。
7．郭北海，张艳敏等。甜菜碱醛脱氢酶(BADH)基因转化小麦及其表达。植物学报。2000，42(3)：279—283。
8．郭光沁。小麦原生质体培养细胞胚胎直接发生再生植株。中国科学(B辑)。1990，9：970—974。
9．葛台明，章荣德等。冬小麦原生质体培养的胚状体直接发生。生物工程学报，2000，16(5)：609—613。
10．何勇刚，林刚等。小麦不同生理状态的幼穗和幼胚盾片与诱导分化能力关系的研究。武汉植物学研究，2001，19(5)：363—368。
11．胡含，张相歧等。花粉小麦染色体工程。科学通报，1999，44(1)：6—11。
12．刘大钧。将簇毛麦种质转移给小麦的研究。遗传学报，1983，10(2)：103—113。
13．李洪杰，朱至清等。组织培养诱导的普通小麦—黑麦代换系和附加系分子细胞遗传学检测。植物学报，1998，40(1)：37—41。
14．李洪杰，郭北海等。利用组织培养与辐射诱变创造高频率小麦与簇毛麦染色体易位，遗传学报，2000，27(6)：511—519。
15．梁辉，郑近等。用基因枪法获得抗白粉病转芪合酶基因小麦。科学通报，1999，44(24)：2644—2648。
16．刘根齐，张孔恬等。外源DNA直接导入小麦及其在育种上的应用。遗传学报，1994，21(6)：463—467。
17．李洪潮，胡道芬等。影响小麦成熟胚培养因素的研究。华北农学报，1990，5(1)：22—27。
18．李洪潮。冬小麦原生质体培养及植株再生。植物学报，1991，33(9)：706—711。
19．李士生，张玉玲。小麦幼穗的组织培养及愈伤组织的分化研究。武汉植物学研究，1990，8(4)：349—354。
20．刘世强，侯秀英等。普通小麦组织培养一步成苗法研究初报。辽宁农业科学，1990，1：53—55。
21．刘文轩，刘大钧。小麦原生质体培养和遗传转化的研究。国外农学—麦类作物，1994，6：34—37。
22．廖祥儒，郭中伟等。低温和PEG预处理对小麦愈伤组织形成及IAA氧化的影响。植物生理学报，2000，17(3)：257—259。
23．刘小冰，刘娜等。蛋白质含量不同的春小麦开花后子粒内源激素的变化。麦类作物学报，2000，20(3)：25—28。
24．刘小冰，曾冰寒等。植物生长调节物质对春小麦种子萌发的效应。东北农学院学报，1991，22(3)：220—224。
25．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。植物生理学通讯，1994，30(4)：255—260。
26．陆瑞菊，陈佩度等。将大赖草种质转移给普通小麦的研究。Ⅳ，通过花药培养选育双重二倍体附加系和代换—附加系。南京农业大学学报，1995，18(4)：1-6。
27．任正隆。八倍体小黑麦×普通小麦杂种后代群体中的染色体易位。遗传学报，1991，18(3)：228—234。
28．任正隆，张怀琼。小麦—黑麦染色体小片段易位的诱导。中国科学C辑，1997，27(3)：258—263。
29．任延国．小麦幼穗愈伤组织原生质体培养再生植株。科学通报，1989，34(9)693—695。
30．孙宝林。提高小麦原生质体再生植株频率的研究。生物工程学报，1990，6：116—19。
31．王海波。小麦原生质体培养—高频率的细胞团形成辑植株再生。中国科学(B辑)，1989，8：828—835。
32．王丽莉，贾敬芬等。影响小麦原生质体培养因素的研究。中国科学技术大学学报，1996，26(4)：505—509。
33．王瑞英。稻麦及豆科作物子粒发育过程中的内源激素水平变化。麦类作物，1997，17(3)：44—47。
34．王小军，刘玉乐等。可育的抗除草剂溴苯氰转基因小麦。植物学报，1996，38(12)：942—948。
35．翁跃进。普通小麦—顶芒山羊草异附加系的创造和鉴定。Ⅰ，小麦花药培养对创建普通小麦—顶芒山羊草异源附加系的作用。作物学报，1995，21(1)：39—44。
36．吴琴生，刘宝等。“中国春”小麦原生质体培养玉形态分化。南京农业大学学报，1993，16(2)：1—6。
37．肖海林，赵世绪。影响小麦幼穗离体培养几个重要因素的研究。北京农业大学学报，1989，15(2)：147—151。
38．肖兴国，张爱民等。转基因小麦的研究进展与展望。农业生物技术学报，2000，8(2)：111—116。
39．许智宏．植物细胞的分化。植物生理学通报，1979，3：66—75。
40．徐琼芳，李连城等。遗传。用天花粉蛋白基因转化小麦获得转基因植株。2001，23(2)：135—137。
41．于晓红，朱祯等。基因枪法转化小麦及转基因植株的获得。高技术通讯，2000，10(2)：13—17。
42．于晓红，朱祯等。提高小麦愈伤组织分化频率的因素。植物生理学报，1999，25(1)：388—394。
43．张桂芳，刘建文等。普通小麦于东方旱麦草异附加系和代换系的选育与原位杂交检测。遗传学报，2000，27(1)：50—55。
44．张晓东，李冬梅等。利用基因枪将HMW谷蛋白亚基基因与除草剂Basta抗性基因导入小麦不同外植体获得转基因植株。遗传，1998，20(增刊)：3—8。
45．张根发。小麦成熟胚与其它外植体再生能力的研究。东北师大学报，1990，8(8)：105—111。
46．张静兰，徐桂芳等。吲哚乙酸、吲哚乙酸氧化酶和几种愈伤组织生长的关系。植物生理学报，1979，5(3)：193—198。
47．郑济芳，王晓萍等。利用Ti质粒介导的二元载体法向小麦导入几丁质酶基因的初步研究。哈尔滨师范大学自然科学学报，1999，15(2)：85—89。
48．曾君祉，王东江。用花粉管途径获得小麦转基因植株。中国科学(B辑)，1993，23(3)：256—261。
49．周文麟，倪建福等。外源C4作物DNA导入小麦的研究。作物学报，1992，18(6)：418—423。
50．周淼平。低浓度2，4-D及外源激素诱导小麦体细胞快速成苗。江苏农业学报，2000，16(3)：139—142。
51. Ahmad,H.zhong, etal.Shoot apical meristem:in vitro regeneration and morphogenesis in wheat(Triticum aestivumc).In vitro Cellular and Developmental Biology Plant,2002,38(2): 163—167.
52. Bennici,L.Caffaro,etal.Callus formation and plantlet regeneration from immature Triticum durum
Defs embryos.Euphytica, 1988,39(1 ):255-263．
53． K.Agawal,Sharad Tiwari,etal.Effect of genotypes and nutrient media on immature embryo culture f wheat.Indian Journal of Genetics and Plant Breeding,1995,55(1) :50-70．
54． Ahmed.KZ,Sagi.F,etal.Protoplast isolation.culture and plantregeneration in wheat and other cereal crops:review and update.Acta Agronomica Hungarica,1995,43(3-4) :371--383．
55． ALC.Dornelles,FIF.Carvalho,etal.Callus induction and plant regeneration by Brazilian triticale and wheat genotypes.Brazilian Journal of Genetics, 1997,20(1 ):41-44．
56． B.Barnabas,E.Szakacs,etal.In vitro androgenesis of wheat:from fundamentals to pratical application. Euphytica,2001,119:211-216．
57． B.Liu,etal.A preliminary study on the relationship between callus growth rate and plantlet regeneration capacity of wheat. Acta Agriculturae Universitatis Jilinensis,1988,10(2) :103-104．
58． Cheng Zengshu,Gao Zhiqiang.etal.High frequency cell division and callus formation in the culture of protoplasts derived from embryogenic callus of wheat. Acta Agriculturae Borealisinica,1994, 9(1) :1-6．
59． EL.Sherbeny,S.Sato,etal.Effect of Genotype and 2,4-D concentration on callus induction from immature embryos of morden Egyption wheat cultivars.Cereal Research Communications,2001, 29(3-4) :305-311．
60． F.Delporte,O.Mostade,etal.Plant regeration through callus inition from thin mature embryo fragments of wheat.Plant Cell Tissue and Organ Culture,2001,1(67) :73-80．
61． J.F.O'HARA.H.E.STREET. Wheat callus culture:the Initiation,Growth and organogenesis of callus derived from various explant sources.Ann.Bot, 1978,42:1029-1038．
62． Harris R, Wright M.etal.Callus for atio and plantlet resneration from protoplant derived from suspension cultures of wheatPlant Cell Rep,1988,7:337-340．
63． He.D.G.J.W.Ouyang.Callus and plantlet formation from cultured wheat anthers at different developmental stages-Plant Sci Lett,1984,33:71-79．
64． He.GY.PA.Lazzeri.Improvement of somatic embryogenesis and plant regeneration from durum wheat scutellum and inflorescence cultures.Euphytica.2001,119(3) :369-376． .
65． HK.Khanna,GE.Daggard.Enhanced Shoot regeneration in nine Australian wheat cultivars by spermidine and water stress treatments-Australian Journal of Plant Physiology,20001,28(12) : 1243-1247．
66． Hou bingkai.Yu Huimin.etal.Effects of low temperature on induction and differentiation of wheat calluses.Plant Cell Tissueand Organ Culture, 1997,49(1) :35-38．
67． I.Ewis Anshulika,etal.Regeneration response from different explant of high yielding genotypes of wheat(Triticum Aestivuml).Crop Improvement,1999,26(1) :5-9．
68． Ingo Potrykus.Micro-targeting of microprojectiles to target areas in the micrometre range.Nature, 1992,355(2) :568-569．
69． J.C.CHIN,K.J.Scott.Studies on the formation of roots and shoots in wheat callus culltures.Ann.Bot, 1976,41:473-481．
70． Jing,Z.Y.Xi,etal.Effects of high temperature and physiological condition of donor plants on induction of pollen derived plants in wheatannRep Inst Genet Sin.Beijing.PP,67-68．
71． J.M.Gonzalez,E.Friero,etal.Influence of genotype and culture medium on callus fonnation and plant regeneration from immature embryos of triticum turgidum desf cultivars.Plant Breeding,2001,20(6) :513-517．
72． Kasem Zahmed.Tibor Bartok,etal.A Modified method for rapid callus induction by utilization of endosperm metabolites in mature and immature seeds of bread wheat and durum wheatCereal Research Communications, 1992,20(1-2) :81-86．
73． Kazuo Sasaki,Koichiro Shimomura etal.IAA metabolism in embryogenic and non-embryogenic carrot cell.Plant CellPhysiol, 1994,35(8) :1159-1164．
74． I.Karsai,Z.Bedo.Effect of carbohydrate content on the embryoid and plant production in triticale anther culture.Cereal Research Communications, 1997,25(2) : 109-116．
75． K.Rajyalakshmi,SK.Dhir,etal.Callusing and regeneratin of plantlets via somatic embryogenesis from inflorescence cultures of Triticum aestivum;role of genotype and long term retention of morphogenic potential.Plant Breeding,1988,101(1) :80-85．
76． S.Kintzios, M.Triantafyllou, etal.Effect of genotype and different growth regulator treatments on callus induction,proliferation and plant regeration from mature wheat embryos.Cereal Research Communitions,1996,24(2) :147-153．
77． Lazar.M.D,P.S>Baenzinger,etal.Combining abilities and heritability of cal;lus formation and plantlet regeneration in wheat(Triticum aestivuml) anther cultures.Theor Appl.Genet, 1984,68:131-134．
78． Lech Michalczuk,David M,etal.Regulation of Indole-3-Acetic Acid biosynthetic pathways in carrot
cell cultures.Plant Physiol, 1992,100:1346-1353．
79． Li Yi-Wem,SEO Yong-Weon,etal.Genetic Transformation in Triticeae crops.Acta Botanica Sinica,2002,44(5) :505-518．
80． Lu Tiegang,Sun Jingsan,etal.Haploid plant regenerationfrom protoplast cultures of durum wheat.Acta Botanica Sinica,1995,37(9) :704-710．
81． Lu.WZ,She.JM,etal.Tissue culture of young spike,stem and node of wheat and plantlet regeneratin.Jiangsu Journal of Agriculture Sciences, 1988,4(1) : 14-18．
82． LV.Prilyuk.Plant regeneration in callus culture of four wheat species.Biologiya Kultiviruemykh kletoki biotekhnologiya,[edited by Butenko.R.G.J],1988,113．
83． Malik M Rafi,Roherts Zemetra,etal.Effect of ABSCISIC ACID on wheat callus culture.Cereal Research Communications,1995,23(4) :375-382．
84． M.Cialacu,l.Hagima,etal.Characterization of somatic embryogenesis in common wheat.Analele Institutului de Cercetari Pentru Cereale Si Plante Tehnice, 1996,63:239-248．
85． M.Bliffeld,J.Mundy,etal.Genetic engineering of wheat for increased risistance to powdery mildew disease.Theor Appl Genet,1998,98:1079-1086．
86． Meenakshi Sidana,S.Uppal ,etal.In vitro regenreation of plants from mature embryos derived cultures of substitution lines in wheat.Annals of Biology Ludhiana,1995,11(1-2) :43-46．
87． MG.Mendoza,HF.Kaeppler.Auxin and sugar effects on callus induction and plant regeneration frequencies from mature embryos of wheat. In Vitro Cellar and Developmental Biologr Plant,2002,38(1) :39-45．
88． Ming Cheng,Joyce E.Fry.etal.Genitic transformation of wheat mediated by agrobacterium tumefaciens.Plant Physiol.1997,115:971-980．
89． Muhesh Dahyia,S.Uppal,etal.Plant regeneration from immature embryos and somaclonal variation in wheatlndian Journal of Plant Physiology, 1995,38(1) : 10-14．
90． M.Ozgen,M.Turet,etal.Callus induction and plant regeneration from immature and mature embryos of winter durum wheat genotypes.Plant Breeding, 1996,115:455-458．
91． M.Ozgen,M.Turet,etal.Efficient callus induction and plant regeneration from mature embryo culture of winter wheat genotypes,Plant Cell Reports, 1998,18:331-335．
92． NE.Bohorova,WH.Pfeiffer,etaI.Regeneration potential of CIMMYT durum wheat and triticale
varieties from immature embtyos.Plant Breeding,2001,120(4) :291-295．
93． N.Saidi,S.Cherkaoui,etal.Embryo formation and regeneration in Triticum turgidum SSp.durum anther culture.Plant Cell Tissue and Organ culture. 1997,25(2) : 109-116．
94． OC.Maes,RN.Chibbar,etal.Somatic embryogenesis from isolated scutella of wheat:effects of physical.physiological and genetic factors.Plant Breeding2000,27(11) :455-458．
95． Ouyang.Y. W,O.G.He,eial.The response of anther culture to culture temperature varies with growth condition of anther donor plants .Plant Sci, 1987,49:145-148．
96． Ouyang,Y.W,C.C.Hu,etal.Induction of pollen plants from anther of Triticum aestivum L.cultureed in vitro.Sci Sin,1973,16:79-95．
97． P.Barcelo,PA.Lazzeri,etal.Competence of cereal leaf cells.Plant Science Limerick,1991,77(2) :243-251．
98． R.G.Sears,E.L.Dekard.Tissue culture variability in wheatxallus induction and plant regeneration. Crop Ssience,1982,22:546-550．
99． Riley.Introduction of yellow rust resistance of Aegilops comosa into wheat by genetically induced homoeologous recombination.Nature, 1986,5126(217) :383-384．
100． R.Tuberosa,S.Ravaglia,etal.Callus induction and plant regneration in Italian cultivars of bread wheat.Agricolture Mediterrance, 1988,118(4) :361-365．
101 . R.Yadava,HS.Chawla,Interaction of genotype.growth regulators and aminoacids on plant regeneration from different developmental stages of inflorescence in wheat.Journal of Genetics and Breeding,2001,55(3) :261-267．
102． Sagi,B.Barnabas.Evidence for cytoplasmic control of in vitro microspore embryogenesis in the anther cultureof wheat.Theor Appl Genet, 1989,78:867-872．
103． SE.Morozova.Duration of regeneration ability in wheat callus.Biologiya Kultiviruemykh Kletok I Biotekhnologiya,[edited by R.Butenko], 1988,135-136．
104． S.Ito,J.Abe.callus formation and plant regeneration from immature wheat embryos.Bulletin of the Tohoku National Agricultural Experiment Station, 1990,81:33-40．
105． S.Fennell,NE.Bohorova,etal.Plant regeneration from immature embryos of 48 elite CIMMYT bread wheat Theo Appl Genet, 1996,92(2) : 163-169．
106． Skoog,F.Armstrang.Cytokinin.Ann Rev Plant Physiol, 1970,21:359-384．
107． S.Sesek,A.Kondic.Genotypic specificity of green plants regenerayion in in vitro wheat anther
culture. Selekcija I Semenarstvo,1996,3(3-4) :23-26．
108． Stober.A.Hess.D.Spike pretreatments,anther culture conditions and anther culture response of 17 German varieties of spring wheat.Plant Breeding, 1997,116(5) :443-447．
109． Szakacs.E,B.Barnabas.Cytoplasmic control of in vitro microspore embryogenesis in the anther culture of wheat.Theor Appl Genet, 1989,78:867-872．
110． S.Kintzios,M.Triantafyllou,etal.Effect of genotype and different growth regulator treatments on callus induction,proliferation and plant regeneration from mature wheat embryos.Cereal Research Communications, 1996,24(2) : 147-153．
111． Takiko Shimada.Plant regeneration from the callus induced from wheat embryo.Japan.J.Genetics, 1978,53(5) :371-374．
112． Tiber Bartok,Ferenc Sagi.etal.A new endospermsupported callus induction method for wheat.Plant Cell Tissue and Ogan Culture,1990,22:37-41．
113． Troy Week,Olin D.Anderson,etal.Rapid production of multiple independent lines of fertile transgenic wheat.Plant Physiol.l993,102:1077-1084．
114． V.K.SharmaA-Rao,etal.Camparison of developmental stages of inflorescence for high frequency plant regenreation in Triticum aestivum Land T.durum desf.Plant Cell Reports, 1995,15(3-4) :227-231．
115． V.K.Tiwari,A.M.Botha,etal.Anew technique for the propagation of somatic wheat embryos. Biotechnology Techniques, 1997,11 (9) :623-636．
116． VM.Jimenez,F.Bangerth.Endogenoushormone concentrations and organ culture,2001,67(1) :37-46．
117． V.Varshney,T.Kant,et al.High frequency plant regeneration from immature embryo cultures of Triticum aestivum and T.Durum.Cereal Research Communications, 1996,24(4) :409-416．
118． Wang.P,Y.R.Chen.Effects lof growth conditions of anther donor plants on the production of pollen plants in wheat anther culture.Acta Genet Sin,Beijing, 1980,7:64-71．
119． Ward.KA,Jordan.MC.Callus formation and plant regeneration from immature and mature embryos of rye.In Vitro Cellular and Developmental Bio;ogy Plant,2001,37(3) :361-368．
120． Xia Guangmin,Li Zhongvi,etal.Transgenic plant regeneration from wheat mediated by Agrobacterium tumefacieus.Acta Phytophysiologica Sinica,1999,25(1) :22-28．
121． Zhang.LJ,P.Seilleur.A simple and fast method to obtain high frequency of plant regeneration from mature and immature wheat embryos.Bulletin des Recherches Agronomiques de Gembioux, 1987,22
(3) :187-197．
122． Zheng.QC,Zhu YL.etal.Plantlet regeneration from in vitro culture of young spikes of wheat and its vayiation.Acta Agriculture Nucleatae Sinica, 1989,3(3) : 129-136．
123． Yang.F,Li>BJ,etal.Factors influencing tissue culture of matured wheat embryo.Guang Dong Agricultural Sciences, 1995,6:12-14．
2．催凯荣，小麦体细胞胚发生中DNA、RNA和蛋白质的合成动态。核农学报，1997，11(4)：209—214。
3．程在全，王芳等。提高小麦体细胞诱导出胚性愈伤组织的方法研究。西南农业学报，1999，12(3)：4—8。
4．陈以峰，汤日圣。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—477。
5．成卓敏，吴茂森等。应用基因枪法获得抗大麦黄矮病毒转基因植株。植物病理学报，2000，30(2)：116—121。
6．关畅，曾寒冰等。小麦未熟胚离体培养的研究。东北农学院学报，1993，24(2)：107—116。
7．郭北海，张艳敏等。甜菜碱醛脱氢酶(BADH)基因转化小麦及其表达。植物学报。2000，42(3)：279—283。
8．郭光沁。小麦原生质体培养细胞胚胎直接发生再生植株。中国科学(B辑)。1990，9：970—974。
9．葛台明，章荣德等。冬小麦原生质体培养的胚状体直接发生。生物工程学报，2000，16(5)：609—613。
10．何勇刚，林刚等。小麦不同生理状态的幼穗和幼胚盾片与诱导分化能力关系的研究。武汉植物学研究，2001，19(5)：363—368。
11．胡含，张相歧等。花粉小麦染色体工程。科学通报，1999，44(1)：6—11。
12．刘大钧。将簇毛麦种质转移给小麦的研究。遗传学报，1983，10(2)：103—113。
13．李洪杰，朱至清等。组织培养诱导的普通小麦—黑麦代换系和附加系分子细胞遗传学检测。植物学报，1998，40(1)：37—41。
14．李洪杰，郭北海等。利用组织培养与辐射诱变创造高频率小麦与簇毛麦染色体易位，遗传学报，2000，27(6)：511—519。
15．梁辉，郑近等。用基因枪法获得抗白粉病转芪合酶基因小麦。科学通报，1999，44(24)：2644—2648。
16．刘根齐，张孔恬等。外源DNA直接导入小麦及其在育种上的应用。遗传学报，1994，21(6)：463—467。
17．李洪潮，胡道芬等。影响小麦成熟胚培养因素的研究。华北农学报，1990，5(1)：22—27。
18．李洪潮。冬小麦原生质体培养及植株再生。植物学报，1991，33(9)：706—711。
19．李士生，张玉玲。小麦幼穗的组织培养及愈伤组织的分化研究。武汉植物学研究，1990，8(4)：349—354。
20．刘世强，侯秀英等。普通小麦组织培养一步成苗法研究初报。辽宁农业科学，1990，1：53—55。
21．刘文轩，刘大钧。小麦原生质体培养和遗传转化的研究。国外农学—麦类作物，1994，6：34—37。
22．廖祥儒，郭中伟等。低温和PEG预处理对小麦愈伤组织形成及IAA氧化的影响。植物生理学报，2000，17(3)：257—259。
23．刘小冰，刘娜等。蛋白质含量不同的春小麦开花后子粒内源激素的变化。麦类作物学报，2000，20(3)：25—28。
24．刘小冰，曾冰寒等。植物生长调节物质对春小麦种子萌发的效应。东北农学院学报，1991，22(3)：220—224。
25．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。植物生理学通讯，1994，30(4)：255—260。
26．陆瑞菊，陈佩度等。将大赖草种质转移给普通小麦的研究。Ⅳ，通过花药培养选育双重二倍体附加系和代换—附加系。南京农业大学学报，1995，18(4)：1-6。
27．任正隆。八倍体小黑麦×普通小麦杂种后代群体中的染色体易位。遗传学报，1991，18(3)：228—234。
28．任正隆，张怀琼。小麦—黑麦染色体小片段易位的诱导。中国科学C辑，1997，27(3)：258—263。
29．任延国．小麦幼穗愈伤组织原生质体培养再生植株。科学通报，1989，34(9)693—695。
30．孙宝林。提高小麦原生质体再生植株频率的研究。生物工程学报，1990，6：116—19。
31．王海波。小麦原生质体培养—高频率的细胞团形成辑植株再生。中国科学(B辑)，1989，8：828—835。
32．王丽莉，贾敬芬等。影响小麦原生质体培养因素的研究。中国科学技术大学学报，1996，26(4)：505—509。
33．王瑞英。稻麦及豆科作物子粒发育过程中的内源激素水平变化。麦类作物，1997，17(3)：44—47。
34．王小军，刘玉乐等。可育的抗除草剂溴苯氰转基因小麦。植物学报，1996，38(12)：942—948。
35．翁跃进。普通小麦—顶芒山羊草异附加系的创造和鉴定。Ⅰ，小麦花药培养对创建普通小麦—顶芒山羊草异源附加系的作用。作物学报，1995，21(1)：39—44。
36．吴琴生，刘宝等。“中国春”小麦原生质体培养玉形态分化。南京农业大学学报，1993，16(2)：1—6。
37．肖海林，赵世绪。影响小麦幼穗离体培养几个重要因素的研究。北京农业大学学报，1989，15(2)：147—151。
38．肖兴国，张爱民等。转基因小麦的研究进展与展望。农业生物技术学报，2000，8(2)：111—116。
39．许智宏．植物细胞的分化。植物生理学通报，1979，3：66—75。
40．徐琼芳，李连城等。遗传。用天花粉蛋白基因转化小麦获得转基因植株。2001，23(2)：135—137。
41．于晓红，朱祯等。基因枪法转化小麦及转基因植株的获得。高技术通讯，2000，10(2)：13—17。
42．于晓红，朱祯等。提高小麦愈伤组织分化频率的因素。植物生理学报，1999，25(1)：388—394。
43．张桂芳，刘建文等。普通小麦于东方旱麦草异附加系和代换系的选育与原位杂交检测。遗传学报，2000，27(1)：50—55。
44．张晓东，李冬梅等。利用基因枪将HMW谷蛋白亚基基因与除草剂Basta抗性基因导入小麦不同外植体获得转基因植株。遗传，1998，20(增刊)：3—8。
45．张根发。小麦成熟胚与其它外植体再生能力的研究。东北师大学报，1990，8(8)：105—111。
46．张静兰，徐桂芳等。吲哚乙酸、吲哚乙酸氧化酶和几种愈伤组织生长的关系。植物生理学报，1979，5(3)：193—198。
47．郑济芳，王晓萍等。利用Ti质粒介导的二元载体法向小麦导入几丁质酶基因的初步研究。哈尔滨师范大学自然科学学报，1999，15(2)：85—89。
48．曾君祉，王东江。用花粉管途径获得小麦转基因植株。中国科学(B辑)，1993，23(3)：256—261。
49．周文麟，倪建福等。外源C4作物DNA导入小麦的研究。作物学报，1992，18(6)：418—423。
50．周淼平。低浓度2，4-D及外源激素诱导小麦体细胞快速成苗。江苏农业学报，2000，16(3)：139—142。
51. Ahmad,H.zhong, etal.Shoot apical meristem:in vitro regeneration and morphogenesis in wheat(Triticum aestivumc).In vitro Cellular and Developmental Biology Plant,2002,38(2): 163—167.
52. Bennici,L.Caffaro,etal.Callus formation and plantlet regeneration from immature Triticum durum
Defs embryos.Euphytica, 1988,39(1 ):255-263．
53． K.Agawal,Sharad Tiwari,etal.Effect of genotypes and nutrient media on immature embryo culture f wheat.Indian Journal of Genetics and Plant Breeding,1995,55(1) :50-70．
54． Ahmed.KZ,Sagi.F,etal.Protoplast isolation.culture and plantregeneration in wheat and other cereal crops:review and update.Acta Agronomica Hungarica,1995,43(3-4) :371--383．
55． ALC.Dornelles,FIF.Carvalho,etal.Callus induction and plant regeneration by Brazilian triticale and wheat genotypes.Brazilian Journal of Genetics, 1997,20(1 ):41-44．
56． B.Barnabas,E.Szakacs,etal.In vitro androgenesis of wheat:from fundamentals to pratical application. Euphytica,2001,119:211-216．
57． B.Liu,etal.A preliminary study on the relationship between callus growth rate and plantlet regeneration capacity of wheat. Acta Agriculturae Universitatis Jilinensis,1988,10(2) :103-104．
58． Cheng Zengshu,Gao Zhiqiang.etal.High frequency cell division and callus formation in the culture of protoplasts derived from embryogenic callus of wheat. Acta Agriculturae Borealisinica,1994, 9(1) :1-6．
59． EL.Sherbeny,S.Sato,etal.Effect of Genotype and 2,4-D concentration on callus induction from immature embryos of morden Egyption wheat cultivars.Cereal Research Communications,2001, 29(3-4) :305-311．
60． F.Delporte,O.Mostade,etal.Plant regeration through callus inition from thin mature embryo fragments of wheat.Plant Cell Tissue and Organ Culture,2001,1(67) :73-80．
61． J.F.O'HARA.H.E.STREET. Wheat callus culture:the Initiation,Growth and organogenesis of callus derived from various explant sources.Ann.Bot, 1978,42:1029-1038．
62． Harris R, Wright M.etal.Callus for atio and plantlet resneration from protoplant derived from suspension cultures of wheatPlant Cell Rep,1988,7:337-340．
63． He.D.G.J.W.Ouyang.Callus and plantlet formation from cultured wheat anthers at different developmental stages-Plant Sci Lett,1984,33:71-79．
64． He.GY.PA.Lazzeri.Improvement of somatic embryogenesis and plant regeneration from durum wheat scutellum and inflorescence cultures.Euphytica.2001,119(3) :369-376． .
65． HK.Khanna,GE.Daggard.Enhanced Shoot regeneration in nine Australian wheat cultivars by spermidine and water stress treatments-Australian Journal of Plant Physiology,20001,28(12) : 1243-1247．
66． Hou bingkai.Yu Huimin.etal.Effects of low temperature on induction and differentiation of wheat calluses.Plant Cell Tissueand Organ Culture, 1997,49(1) :35-38．
67． I.Ewis Anshulika,etal.Regeneration response from different explant of high yielding genotypes of wheat(Triticum Aestivuml).Crop Improvement,1999,26(1) :5-9．
68． Ingo Potrykus.Micro-targeting of microprojectiles to target areas in the micrometre range.Nature, 1992,355(2) :568-569．
69． J.C.CHIN,K.J.Scott.Studies on the formation of roots and shoots in wheat callus culltures.Ann.Bot, 1976,41:473-481．
70． Jing,Z.Y.Xi,etal.Effects of high temperature and physiological condition of donor plants on induction of pollen derived plants in wheatannRep Inst Genet Sin.Beijing.PP,67-68．
71． J.M.Gonzalez,E.Friero,etal.Influence of genotype and culture medium on callus fonnation and plant regeneration from immature embryos of triticum turgidum desf cultivars.Plant Breeding,2001,20(6) :513-517．
72． Kasem Zahmed.Tibor Bartok,etal.A Modified method for rapid callus induction by utilization of endosperm metabolites in mature and immature seeds of bread wheat and durum wheatCereal Research Communications, 1992,20(1-2) :81-86．
73． Kazuo Sasaki,Koichiro Shimomura etal.IAA metabolism in embryogenic and non-embryogenic carrot cell.Plant CellPhysiol, 1994,35(8) :1159-1164．
74． I.Karsai,Z.Bedo.Effect of carbohydrate content on the embryoid and plant production in triticale anther culture.Cereal Research Communications, 1997,25(2) : 109-116．
75． K.Rajyalakshmi,SK.Dhir,etal.Callusing and regeneratin of plantlets via somatic embryogenesis from inflorescence cultures of Triticum aestivum;role of genotype and long term retention of morphogenic potential.Plant Breeding,1988,101(1) :80-85．
76． S.Kintzios, M.Triantafyllou, etal.Effect of genotype and different growth regulator treatments on callus induction,proliferation and plant regeration from mature wheat embryos.Cereal Research Communitions,1996,24(2) :147-153．
77． Lazar.M.D,P.S>Baenzinger,etal.Combining abilities and heritability of cal;lus formation and plantlet regeneration in wheat(Triticum aestivuml) anther cultures.Theor Appl.Genet, 1984,68:131-134．
78． Lech Michalczuk,David M,etal.Regulation of Indole-3-Acetic Acid biosynthetic pathways in carrot
cell cultures.Plant Physiol, 1992,100:1346-1353．
79． Li Yi-Wem,SEO Yong-Weon,etal.Genetic Transformation in Triticeae crops.Acta Botanica Sinica,2002,44(5) :505-518．
80． Lu Tiegang,Sun Jingsan,etal.Haploid plant regenerationfrom protoplast cultures of durum wheat.Acta Botanica Sinica,1995,37(9) :704-710．
81． Lu.WZ,She.JM,etal.Tissue culture of young spike,stem and node of wheat and plantlet regeneratin.Jiangsu Journal of Agriculture Sciences, 1988,4(1) : 14-18．
82． LV.Prilyuk.Plant regeneration in callus culture of four wheat species.Biologiya Kultiviruemykh kletoki biotekhnologiya,[edited by Butenko.R.G.J],1988,113．
83． Malik M Rafi,Roherts Zemetra,etal.Effect of ABSCISIC ACID on wheat callus culture.Cereal Research Communications,1995,23(4) :375-382．
84． M.Cialacu,l.Hagima,etal.Characterization of somatic embryogenesis in common wheat.Analele Institutului de Cercetari Pentru Cereale Si Plante Tehnice, 1996,63:239-248．
85． M.Bliffeld,J.Mundy,etal.Genetic engineering of wheat for increased risistance to powdery mildew disease.Theor Appl Genet,1998,98:1079-1086．
86． Meenakshi Sidana,S.Uppal ,etal.In vitro regenreation of plants from mature embryos derived cultures of substitution lines in wheat.Annals of Biology Ludhiana,1995,11(1-2) :43-46．
87． MG.Mendoza,HF.Kaeppler.Auxin and sugar effects on callus induction and plant regeneration frequencies from mature embryos of wheat. In Vitro Cellar and Developmental Biologr Plant,2002,38(1) :39-45．
88． Ming Cheng,Joyce E.Fry.etal.Genitic transformation of wheat mediated by agrobacterium tumefaciens.Plant Physiol.1997,115:971-980．
89． Muhesh Dahyia,S.Uppal,etal.Plant regeneration from immature embryos and somaclonal variation in wheatlndian Journal of Plant Physiology, 1995,38(1) : 10-14．
90． M.Ozgen,M.Turet,etal.Callus induction and plant regeneration from immature and mature embryos of winter durum wheat genotypes.Plant Breeding, 1996,115:455-458．
91． M.Ozgen,M.Turet,etal.Efficient callus induction and plant regeneration from mature embryo culture of winter wheat genotypes,Plant Cell Reports, 1998,18:331-335．
92． NE.Bohorova,WH.Pfeiffer,etaI.Regeneration potential of CIMMYT durum wheat and triticale
varieties from immature embtyos.Plant Breeding,2001,120(4) :291-295．
93． N.Saidi,S.Cherkaoui,etal.Embryo formation and regeneration in Triticum turgidum SSp.durum anther culture.Plant Cell Tissue and Organ culture. 1997,25(2) : 109-116．
94． OC.Maes,RN.Chibbar,etal.Somatic embryogenesis from isolated scutella of wheat:effects of physical.physiological and genetic factors.Plant Breeding2000,27(11) :455-458．
95． Ouyang.Y. W,O.G.He,eial.The response of anther culture to culture temperature varies with growth condition of anther donor plants .Plant Sci, 1987,49:145-148．
96． Ouyang,Y.W,C.C.Hu,etal.Induction of pollen plants from anther of Triticum aestivum L.cultureed in vitro.Sci Sin,1973,16:79-95．
97． P.Barcelo,PA.Lazzeri,etal.Competence of cereal leaf cells.Plant Science Limerick,1991,77(2) :243-251．
98． R.G.Sears,E.L.Dekard.Tissue culture variability in wheatxallus induction and plant regeneration. Crop Ssience,1982,22:546-550．
99． Riley.Introduction of yellow rust resistance of Aegilops comosa into wheat by genetically induced homoeologous recombination.Nature, 1986,5126(217) :383-384．
100． R.Tuberosa,S.Ravaglia,etal.Callus induction and plant regneration in Italian cultivars of bread wheat.Agricolture Mediterrance, 1988,118(4) :361-365．
101 . R.Yadava,HS.Chawla,Interaction of genotype.growth regulators and aminoacids on plant regeneration from different developmental stages of inflorescence in wheat.Journal of Genetics and Breeding,2001,55(3) :261-267．
102． Sagi,B.Barnabas.Evidence for cytoplasmic control of in vitro microspore embryogenesis in the anther cultureof wheat.Theor Appl Genet, 1989,78:867-872．
103． SE.Morozova.Duration of regeneration ability in wheat callus.Biologiya Kultiviruemykh Kletok I Biotekhnologiya,[edited by R.Butenko], 1988,135-136．
104． S.Ito,J.Abe.callus formation and plant regeneration from immature wheat embryos.Bulletin of the Tohoku National Agricultural Experiment Station, 1990,81:33-40．
105． S.Fennell,NE.Bohorova,etal.Plant regeneration from immature embryos of 48 elite CIMMYT bread wheat Theo Appl Genet, 1996,92(2) : 163-169．
106． Skoog,F.Armstrang.Cytokinin.Ann Rev Plant Physiol, 1970,21:359-384．
107． S.Sesek,A.Kondic.Genotypic specificity of green plants regenerayion in in vitro wheat anther
culture. Selekcija I Semenarstvo,1996,3(3-4) :23-26．
108． Stober.A.Hess.D.Spike pretreatments,anther culture conditions and anther culture response of 17 German varieties of spring wheat.Plant Breeding, 1997,116(5) :443-447．
109． Szakacs.E,B.Barnabas.Cytoplasmic control of in vitro microspore embryogenesis in the anther culture of wheat.Theor Appl Genet, 1989,78:867-872．
110． S.Kintzios,M.Triantafyllou,etal.Effect of genotype and different growth regulator treatments on callus induction,proliferation and plant regeneration from mature wheat embryos.Cereal Research Communications, 1996,24(2) : 147-153．
111． Takiko Shimada.Plant regeneration from the callus induced from wheat embryo.Japan.J.Genetics, 1978,53(5) :371-374．
112． Tiber Bartok,Ferenc Sagi.etal.A new endospermsupported callus induction method for wheat.Plant Cell Tissue and Ogan Culture,1990,22:37-41．
113． Troy Week,Olin D.Anderson,etal.Rapid production of multiple independent lines of fertile transgenic wheat.Plant Physiol.l993,102:1077-1084．
114． V.K.SharmaA-Rao,etal.Camparison of developmental stages of inflorescence for high frequency plant regenreation in Triticum aestivum Land T.durum desf.Plant Cell Reports, 1995,15(3-4) :227-231．
115． V.K.Tiwari,A.M.Botha,etal.Anew technique for the propagation of somatic wheat embryos. Biotechnology Techniques, 1997,11 (9) :623-636．
116． VM.Jimenez,F.Bangerth.Endogenoushormone concentrations and organ culture,2001,67(1) :37-46．
117． V.Varshney,T.Kant,et al.High frequency plant regeneration from immature embryo cultures of Triticum aestivum and T.Durum.Cereal Research Communications, 1996,24(4) :409-416．
118． Wang.P,Y.R.Chen.Effects lof growth conditions of anther donor plants on the production of pollen plants in wheat anther culture.Acta Genet Sin,Beijing, 1980,7:64-71．
119． Ward.KA,Jordan.MC.Callus formation and plant regeneration from immature and mature embryos of rye.In Vitro Cellular and Developmental Bio;ogy Plant,2001,37(3) :361-368．
120． Xia Guangmin,Li Zhongvi,etal.Transgenic plant regeneration from wheat mediated by Agrobacterium tumefacieus.Acta Phytophysiologica Sinica,1999,25(1) :22-28．
121． Zhang.LJ,P.Seilleur.A simple and fast method to obtain high frequency of plant regeneration from mature and immature wheat embryos.Bulletin des Recherches Agronomiques de Gembioux, 1987,22
(3) :187-197．
122． Zheng.QC,Zhu YL.etal.Plantlet regeneration from in vitro culture of young spikes of wheat and its vayiation.Acta Agriculture Nucleatae Sinica, 1989,3(3) : 129-136．
123． Yang.F,Li>BJ,etal.Factors influencing tissue culture of matured wheat embryo.Guang Dong Agricultural Sciences, 1995,6:12-14．