转基因草地早熟禾植株及其子代种子抗旱耐盐性分析
|
摘要
为了验证并比较CMO-BADH双基因、CMO基因和DREB1A基因的抗旱和耐盐性功能,评定各转基因株系抗旱和耐盐能力,本研究通过对非转基因草地早熟禾、转CMO-BADH双基因草地早熟禾、转CMO基因草地早熟禾和转DREB1A基因草地早熟禾进行干旱胁迫试验和不同浓度的NaCl胁迫试验,测定了其叶片相对含水量、细胞膜的透性、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性、氧化氢酶(CAT)活性等,结果表明:CMO-BADH双基因、CMO基因和DREB1A基因的转化均能提高草地早熟禾的抗旱性和耐盐性。本研究还通过测定转基因T1代种子和非转基因商品种在PEG胁迫条件下的相对发芽率,评定了转基因T1代种子和非转基因商品种子的抗旱能力,结果表明:转基因T1代种子的抗旱能力大于非转基因商品种子的抗旱能力。上述研究为草地早熟禾抗旱耐盐新品种的选育提供了理论依据和植物材料。
In this experiment, in order to analysis and compare the functions of CMO-BADH double gene、CMO gene and DREB1A gene, to order the salt tolerance and the drought tolerance of all lines and to analysis the drought tolerance ability of T1 filial generation, the effects of drought resistance and different NaCl concentration stress to the tissue culture of transgenic Kentucky bluegrass with CMO-BADH double gene、CMO gene and DREB1A gene were studied on. And the relation water content (RWC), relative electrical conductivity, MDA content, SOD activity, POD activity and CAT activity were measured. The results showed that the CMO-BADH double gene, CMO gene and DREB1A gene all could enhance the drought and salt tolerance of transgenic Kentucky bluegrass.Through studying the relative germination percentage of T1 filial generation and commercial breed under PEG drought resistance,the drought tolerance of the T1 filial generation and commercial breed was ordered. The results showed that the drought tolerance of T1 filial generation is better than commercial breed. And this study also provided theoretical evidence and plant materials for breeding the new Kentucky bluegrass varieties tolerant to drought and salt.
In this experiment, in order to analysis and compare the functions of CMO-BADH double gene、CMO gene and DREB1A gene, to order the salt tolerance and the drought tolerance of all lines and to analysis the drought tolerance ability of T1 filial generation, the effects of drought resistance and different NaCl concentration stress to the tissue culture of transgenic Kentucky bluegrass with CMO-BADH double gene、CMO gene and DREB1A gene were studied on. And the relation water content (RWC), relative electrical conductivity, MDA content, SOD activity, POD activity and CAT activity were measured. The results showed that the CMO-BADH double gene, CMO gene and DREB1A gene all could enhance the drought and salt tolerance of transgenic Kentucky bluegrass.Through studying the relative germination percentage of T1 filial generation and commercial breed under PEG drought resistance,the drought tolerance of the T1 filial generation and commercial breed was ordered. The results showed that the drought tolerance of T1 filial generation is better than commercial breed. And this study also provided theoretical evidence and plant materials for breeding the new Kentucky bluegrass varieties tolerant to drought and salt.
引文
1 曹致中.肯塔基草地早熟禾坪用性状比较试验[J].甘肃农业大学学报,1996,31(2):121-125
2 陈传芳.通过农杆菌介导法获得耐盐转甜菜碱醛脱氢酶基因白三叶草[D].北京:中国科学院遗传与发育生物学研究所,2003:45-68
3 董丽华.草地早熟禾不同品种抗旱性研究[D].宁夏:宁夏大学,2005:15-18
4 额木和.大青山草地早熟禾[J].内蒙古畜牧科学,1996,31(2):44-45
5 高俊凤.植物生理学实验技术[M].西安:世界图书出版社公司,2000:194-195
6 郭彦军,倪郁,吕俊等.豆科牧草种子萌发特性与其抗早性差异的研究[J].中国草地,2003,25(3):24-27
7 侯德英.转 CMO 和 BADH 基因烟草抗旱性及山杨转基因研究[D].哈尔滨:东北林业大学, 2003:35-48
8 贾庚祥.BADH 基因转化番茄提高耐盐性的研究[D].北京:中国科学院植物研究所,2002:06-08
9 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:164-165
10 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:167-169
11 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:260-261
12 李和平.草地早熟禾胚胎学研究[J].武汉植物学研究,1996,14(1): 25-27
13 李和平.草地早熟禾无融合生殖现象的研究[J]武汉植物学研究,1991,9(l):11-14
14 梁慧敏,夏阳,王太明.植物抗寒冻、抗旱、耐盐基因工程研究进展[J].草业学报,2003,12(3):1-7
15 刘强,张贵友,陈受宜.植物转录因子的结构与调控作用[J].科学通报,2000,45(14):1465-1474
16 牛菊兰.北方冷季型草坪草耐盐能力的测定[J].草地学报,1997,5(3):190-19
17 钱俊芝.覆土处理对草地早熟禾草坪质量的影响[J].中国草地,1999, 5 (1):37-40
18 申晓辉.菠菜胆碱单氧化酶(CMO)基因克隆表达及对北美乔松转化的研究[D].北京:北京林业大学,2002:06-27
19 沈义国,杜保兴,张劲松,陈受宜.山菠菜胆碱单氧化物酶基因(CMO)的克隆与分析[J].生物工程学报,2001,17(1):1-6
20 宋淑明.草坪草叶绿素含量与动态分析[J].中国草地,1989,16(6):21-24
21 特木尔布.苜蓿雄性不育杂交制种在生产上的研究[J].草业科学,1999,16(2):23-27
22 王秋叶,郭志远.化学杀雄效果配方研究[J].陕西农业科学,1998,22(2):15-17
23 王艳青,陈雪梅,李悦.植物抗逆中的渗透调节物质及其转基因工程进展[J].北京林业大学学报,2001,23(4):66-70
24 王忠华,李旭晨,夏英武.作物抗旱的作用机制及其基因工程改良研究进展[J].生物技术通报,2002,22(1):16-19
25 吴相钰,赵微平,匡廷云等.植物生理学补充教材[M].北京:中国植物学会生理学会专业委员会北京植物生理学会,1996: 16-27
26 杨宏远.用整体染色与透明技术观察胚囊、胚、胚乳和胚状体[J].植物学报,1986, 28(6):575-581
27 殷朝珍,王兆龙,葛才林.草地早熟禾无融合生殖及其育种利用研究进展[J].草原与草坪,2006,(1):18-23
28 张丽霞,姚爱兴. 国内外草地早熟禾育种技术及其研究进展[J].宁夏农学院学报,2002, 23 (2):76-79
29 赵桂琴.温汤去雄在早熟禾杂交中的应用[J].草业科学,2000,22(1):23-26
30 赵桂琴.早熟禾育种的研究进展与现状[J].甘肃农业大学学报,2000,26(2):119 一 126
31 赵可夫.作物抗性生理[M].北京:农业出版社,1990:249-313
32 赵世绪.无融合生殖与植物育种[M].北京:北京农业大学出版社,1990: 250-322
33 中国科学院上海植物生理研究所,上海市植物生理学会.现代植物生理学实验指南[M].北京:科学出版社,1999:302-303
34 Bashaw,Hanna. Apomictic reproduction[M]. NEW BRUNSWICK: RUTGERS THE STATE UNIVERSITY OF NEW JERSEY, 1991:222-276
35 Brede,Willard.Registration of Nustar Kentucky bluegrass[J]. Crop Science,1993,33(6):1414-1415
36 David R. Huff Determining genetic origins of aberrantrogeny facultative apomictic Kentucky bluegrass using a combination of flow cytometry and silver-staind RAPD markers[J]. Thero Appl Genet,1993,22(7):201-208
37 Grazi ,Akerberg.Observations on the mode of reproduction and embryology of Poa pratynsis[J]Hereditas,1961, 28(8):489-541
38 Grun. Cytogenetic studies of Poa1 Chromosome numbers and morphology of interspecific hybrids[J].Am Jour Bot,1951,39(9):318-323
39 Hair,J B, Hall.The origin of new chromosomes in Agropyron[J]. Heredity Suppl,1952,(6): 215-233
40 Nitzsche.Interspecific hybrids between apomictic forms of Poa palustris with Poa pratensis [J]. InProc,1983,(8) I55-157
41 Nygrcn.Studies in the inheritance of apomixis in the genus Poa[A]. NEW BRUNSWICK :Cong Genet Proc, 1953:842-843
42 Ovesnov.Grass seed with two embryos[J]. Priroda Leningrad,1956,(11):111-112
43 Pepin&Funk.Interspecific hybridization as a method of breeding Kentucky bluegrass for turf[J]. Crop Sci,1971,(11):445-448
44 Rathinasabapathi B,Burnet M,Russell B L.Choline monooxygenease an unusual iron sulfur enzyme catalzing the first step of glycine betaine synthesis in plants:prosthetic group characte rization and cDNA cloning[J]. Proc Natl Acad Sci,1997,94(11):3454-3458
45 Russell B L,Rathinasabapathi B,Hanson A D.Osomtic stress induces expression of choline monooxygenase in sugar beet and amaranth[J].Plant Physiol,1998,116(2):859-865
46 ShenYG,Du BX,Zhang WK,Zhang JS,Chen SY. AhCMO regulated by stresses in Atriplex hortensis can improve drought tolerance in transgenic tobacco[J]. Theor Appl Genet, 2002,105(2):815–821
47 Yong B A,Bashaw E. Cleared-pistil and thick-sectioning techniques for detecting aposporous apomixes[J]. Can J Bot, 1979,28( 7):1668-1672
48 Zhang,Gengyun .Enhancing salt and drought tolerance of Poa and triploid Cynodon with BADH gene and somaclonal variation[D]. NEW BRUNSWICK :RUTGERS THE STATE UNIVERSITY OF NEW JERSEY,2003:268-372
2 陈传芳.通过农杆菌介导法获得耐盐转甜菜碱醛脱氢酶基因白三叶草[D].北京:中国科学院遗传与发育生物学研究所,2003:45-68
3 董丽华.草地早熟禾不同品种抗旱性研究[D].宁夏:宁夏大学,2005:15-18
4 额木和.大青山草地早熟禾[J].内蒙古畜牧科学,1996,31(2):44-45
5 高俊凤.植物生理学实验技术[M].西安:世界图书出版社公司,2000:194-195
6 郭彦军,倪郁,吕俊等.豆科牧草种子萌发特性与其抗早性差异的研究[J].中国草地,2003,25(3):24-27
7 侯德英.转 CMO 和 BADH 基因烟草抗旱性及山杨转基因研究[D].哈尔滨:东北林业大学, 2003:35-48
8 贾庚祥.BADH 基因转化番茄提高耐盐性的研究[D].北京:中国科学院植物研究所,2002:06-08
9 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:164-165
10 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:167-169
11 李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:260-261
12 李和平.草地早熟禾胚胎学研究[J].武汉植物学研究,1996,14(1): 25-27
13 李和平.草地早熟禾无融合生殖现象的研究[J]武汉植物学研究,1991,9(l):11-14
14 梁慧敏,夏阳,王太明.植物抗寒冻、抗旱、耐盐基因工程研究进展[J].草业学报,2003,12(3):1-7
15 刘强,张贵友,陈受宜.植物转录因子的结构与调控作用[J].科学通报,2000,45(14):1465-1474
16 牛菊兰.北方冷季型草坪草耐盐能力的测定[J].草地学报,1997,5(3):190-19
17 钱俊芝.覆土处理对草地早熟禾草坪质量的影响[J].中国草地,1999, 5 (1):37-40
18 申晓辉.菠菜胆碱单氧化酶(CMO)基因克隆表达及对北美乔松转化的研究[D].北京:北京林业大学,2002:06-27
19 沈义国,杜保兴,张劲松,陈受宜.山菠菜胆碱单氧化物酶基因(CMO)的克隆与分析[J].生物工程学报,2001,17(1):1-6
20 宋淑明.草坪草叶绿素含量与动态分析[J].中国草地,1989,16(6):21-24
21 特木尔布.苜蓿雄性不育杂交制种在生产上的研究[J].草业科学,1999,16(2):23-27
22 王秋叶,郭志远.化学杀雄效果配方研究[J].陕西农业科学,1998,22(2):15-17
23 王艳青,陈雪梅,李悦.植物抗逆中的渗透调节物质及其转基因工程进展[J].北京林业大学学报,2001,23(4):66-70
24 王忠华,李旭晨,夏英武.作物抗旱的作用机制及其基因工程改良研究进展[J].生物技术通报,2002,22(1):16-19
25 吴相钰,赵微平,匡廷云等.植物生理学补充教材[M].北京:中国植物学会生理学会专业委员会北京植物生理学会,1996: 16-27
26 杨宏远.用整体染色与透明技术观察胚囊、胚、胚乳和胚状体[J].植物学报,1986, 28(6):575-581
27 殷朝珍,王兆龙,葛才林.草地早熟禾无融合生殖及其育种利用研究进展[J].草原与草坪,2006,(1):18-23
28 张丽霞,姚爱兴. 国内外草地早熟禾育种技术及其研究进展[J].宁夏农学院学报,2002, 23 (2):76-79
29 赵桂琴.温汤去雄在早熟禾杂交中的应用[J].草业科学,2000,22(1):23-26
30 赵桂琴.早熟禾育种的研究进展与现状[J].甘肃农业大学学报,2000,26(2):119 一 126
31 赵可夫.作物抗性生理[M].北京:农业出版社,1990:249-313
32 赵世绪.无融合生殖与植物育种[M].北京:北京农业大学出版社,1990: 250-322
33 中国科学院上海植物生理研究所,上海市植物生理学会.现代植物生理学实验指南[M].北京:科学出版社,1999:302-303
34 Bashaw,Hanna. Apomictic reproduction[M]. NEW BRUNSWICK: RUTGERS THE STATE UNIVERSITY OF NEW JERSEY, 1991:222-276
35 Brede,Willard.Registration of Nustar Kentucky bluegrass[J]. Crop Science,1993,33(6):1414-1415
36 David R. Huff Determining genetic origins of aberrantrogeny facultative apomictic Kentucky bluegrass using a combination of flow cytometry and silver-staind RAPD markers[J]. Thero Appl Genet,1993,22(7):201-208
37 Grazi ,Akerberg.Observations on the mode of reproduction and embryology of Poa pratynsis[J]Hereditas,1961, 28(8):489-541
38 Grun. Cytogenetic studies of Poa1 Chromosome numbers and morphology of interspecific hybrids[J].Am Jour Bot,1951,39(9):318-323
39 Hair,J B, Hall.The origin of new chromosomes in Agropyron[J]. Heredity Suppl,1952,(6): 215-233
40 Nitzsche.Interspecific hybrids between apomictic forms of Poa palustris with Poa pratensis [J]. InProc,1983,(8) I55-157
41 Nygrcn.Studies in the inheritance of apomixis in the genus Poa[A]. NEW BRUNSWICK :Cong Genet Proc, 1953:842-843
42 Ovesnov.Grass seed with two embryos[J]. Priroda Leningrad,1956,(11):111-112
43 Pepin&Funk.Interspecific hybridization as a method of breeding Kentucky bluegrass for turf[J]. Crop Sci,1971,(11):445-448
44 Rathinasabapathi B,Burnet M,Russell B L.Choline monooxygenease an unusual iron sulfur enzyme catalzing the first step of glycine betaine synthesis in plants:prosthetic group characte rization and cDNA cloning[J]. Proc Natl Acad Sci,1997,94(11):3454-3458
45 Russell B L,Rathinasabapathi B,Hanson A D.Osomtic stress induces expression of choline monooxygenase in sugar beet and amaranth[J].Plant Physiol,1998,116(2):859-865
46 ShenYG,Du BX,Zhang WK,Zhang JS,Chen SY. AhCMO regulated by stresses in Atriplex hortensis can improve drought tolerance in transgenic tobacco[J]. Theor Appl Genet, 2002,105(2):815–821
47 Yong B A,Bashaw E. Cleared-pistil and thick-sectioning techniques for detecting aposporous apomixes[J]. Can J Bot, 1979,28( 7):1668-1672
48 Zhang,Gengyun .Enhancing salt and drought tolerance of Poa and triploid Cynodon with BADH gene and somaclonal variation[D]. NEW BRUNSWICK :RUTGERS THE STATE UNIVERSITY OF NEW JERSEY,2003:268-372