甘蓝型油菜小孢子离体EMS诱变技术研究
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摘要
甲基磺酸乙酯(Ethl methane sulfonste,EMS)是一种能够诱导染色体结构变异的烷化剂,已被广泛应用于诱变育种和种质创新。本实验采用不同EMS浓度(0.02%、0.04%、0.08%(v/v))和处理时间(10min和20min)对7679-1、7679-2、7680、7681和2145在内的5个品系的甘蓝型油菜小孢子进行诱变处理,对诱变后小孢子的胚胎发生能力、植株再生能力、再生植株形态和种子品质变异等进行了研究。主要研究结果如下:
1.不同基因型小孢子胚胎发生能力均存在明显差异。品系7679胚胎发生能力达到100%,2050-2等材料胚胎发生能力低,产胚率0—1.7个/蕾。同一品系不同株系之间胚胎发生能力无显著差异,如品系2050的4个株系的单株胚胎发生能力在0—4.0%之间。
2.不同EMS浓度和处理时间对小孢子产胚率的影响。在EMS浓度相同时,材料7679-2、7680小孢子产胚数随诱变时间延长而减少;在处理时间相同时,材料7679-2、7680小孢子产胚数随诱变剂浓度增加而减少,且随着诱变剂浓度增大产胚数下降速度加快。
3.确定不同基因型材料的EMS诱变半致死剂量。7679-1处理10min时的半致死剂量为0.04%;7679-2处理10-20 min时的半致死剂量为0.08%;7680处理20min时的半致死剂量0.04%。
4.获得了不同表型的突变体。经济性状突变:矮化、短角果、分枝数增多等突变体;花器官突变:粉色花瓣、白色花瓣、雄蕊多于6枚、花瓣数增加、柱头外露等突变体;种子品质突变:获得了含油量43.81%的植株(供体植株含油量38.12-40.93%),油酸含量68.64%的DH植株(供体植株油酸含量62.93-65.82%)。。
5.EMS诱变对小孢子染色体加倍的影响。7679-2未诱变处理的小孢子经秋水仙碱处理后再生植株加倍率54.0%,EMS诱变后经秋水仙碱处理再生植株加倍率在50.0-92.4%,大部分高于未诱变处理;7680未诱变处理的小孢子经秋水仙碱处理的加倍率96.7%,EMS诱变后经秋水仙碱处理再生植株加倍率在85.7-98.0%,EMS诱变对再生植株染色体加倍效果有影响。
Ethl methane sulfonste(EMS) is a kind of alkylating agent which has remarkable mutagenesis effect in chromosome mutagenesis. It has been widely applied in auxiliary breeding and germplasm innovation. This experiment adopts three EMS concentrationsnamely 0.02%, 0.04% and 0.08% (v/v) and two treating durations which are 10 minutes and 20 minutes to treat on in vitro microspore of nine lines from five strains in Brassica napus ,that including 7679-1、7679-2、7680. Investigate the microspore's embryogenesis ,regenerative ability, and the microspore-derived plant's phenotypic and qualitive variation. The finding results are listed as follows:
1. There are diffences in microspore embryogenesis of various genotypes, material7679 get embryo easily,the ability of embryogenesis can reach 100%. 2050-2 embryogenesis is very low, which produces 0-1.7 embryoes per alabastrum. Embryogenesis ability tends to be consistent in different lines of the same strain. For example the embryogenesis ability are all extremely low as 0-4.0% in fore lines of strain 2050.
2. The influence of EMS concentration and treating duration on microspore-yield embryo number . when EMS concentration is same every material's embryo number are always reduced by prolong of treatment duration and the longer the time is the speed ofembryo generation descends slower..when treating duration is same 7679-2、7680'sembryo number reduced accelerative by increase EMS concentration, and the more the concentration is the speed of embryo generation descends faster..
3. Every material has its own medial lethal concentration(LC5o): 7679-1 producing embryoes was about half of the CK when EMS concentration was 0.04% and the duration of EMS treatment was 10min; 7679-2 producing embryoes was about half of the CK when EMS concentration was 0.08% and the duration of EMS treatment was 10-20min;7680 producing embryoes was about half of the CK when EMS concentration was 0.04% and the duration of EMS treatment was 20min.
4. The phenotypic variation has get from DH plants' by EMS mutagenesis. Variation in economic character : height decreases, silique becomes shorter , branching number increases. Flower organ morphological variations including : pink petal, white peta; got more than six stamens, more than fore petalsl, and stigma exsertion. Qualitive variation:got a DH plant whose oil content achieved 43.81% (Donor adult plant index of oil 38.12-40.93%), a DH plant whose oleic acid content was 68.64% (Donor adult plant oleic acid content 62.93-65.82%).
5. EMS treatment influence the chromosome double rate of microspore-derived plants. In 7679-2's regeneration group, CK chromosome double rate was 54.0% by treating with colchicines, regeneration groups which treated by EMS before colchicines treating that chromosome double rate were 50.0-92.4%, were mostly higher than CK. In 7680's regeneration group, CK chromosome double rate was 96.7% by treating with colchicines; regeneration group which treated by EMS before colchicines treating that chromosome double rate were 85.7-98.0%. In a certain amount of concentration, EMS mutagenesis treatment approved the double of regeneration plant chromosome.
1.不同基因型小孢子胚胎发生能力均存在明显差异。品系7679胚胎发生能力达到100%,2050-2等材料胚胎发生能力低,产胚率0—1.7个/蕾。同一品系不同株系之间胚胎发生能力无显著差异,如品系2050的4个株系的单株胚胎发生能力在0—4.0%之间。
2.不同EMS浓度和处理时间对小孢子产胚率的影响。在EMS浓度相同时,材料7679-2、7680小孢子产胚数随诱变时间延长而减少;在处理时间相同时,材料7679-2、7680小孢子产胚数随诱变剂浓度增加而减少,且随着诱变剂浓度增大产胚数下降速度加快。
3.确定不同基因型材料的EMS诱变半致死剂量。7679-1处理10min时的半致死剂量为0.04%;7679-2处理10-20 min时的半致死剂量为0.08%;7680处理20min时的半致死剂量0.04%。
4.获得了不同表型的突变体。经济性状突变:矮化、短角果、分枝数增多等突变体;花器官突变:粉色花瓣、白色花瓣、雄蕊多于6枚、花瓣数增加、柱头外露等突变体;种子品质突变:获得了含油量43.81%的植株(供体植株含油量38.12-40.93%),油酸含量68.64%的DH植株(供体植株油酸含量62.93-65.82%)。。
5.EMS诱变对小孢子染色体加倍的影响。7679-2未诱变处理的小孢子经秋水仙碱处理后再生植株加倍率54.0%,EMS诱变后经秋水仙碱处理再生植株加倍率在50.0-92.4%,大部分高于未诱变处理;7680未诱变处理的小孢子经秋水仙碱处理的加倍率96.7%,EMS诱变后经秋水仙碱处理再生植株加倍率在85.7-98.0%,EMS诱变对再生植株染色体加倍效果有影响。
Ethl methane sulfonste(EMS) is a kind of alkylating agent which has remarkable mutagenesis effect in chromosome mutagenesis. It has been widely applied in auxiliary breeding and germplasm innovation. This experiment adopts three EMS concentrationsnamely 0.02%, 0.04% and 0.08% (v/v) and two treating durations which are 10 minutes and 20 minutes to treat on in vitro microspore of nine lines from five strains in Brassica napus ,that including 7679-1、7679-2、7680. Investigate the microspore's embryogenesis ,regenerative ability, and the microspore-derived plant's phenotypic and qualitive variation. The finding results are listed as follows:
1. There are diffences in microspore embryogenesis of various genotypes, material7679 get embryo easily,the ability of embryogenesis can reach 100%. 2050-2 embryogenesis is very low, which produces 0-1.7 embryoes per alabastrum. Embryogenesis ability tends to be consistent in different lines of the same strain. For example the embryogenesis ability are all extremely low as 0-4.0% in fore lines of strain 2050.
2. The influence of EMS concentration and treating duration on microspore-yield embryo number . when EMS concentration is same every material's embryo number are always reduced by prolong of treatment duration and the longer the time is the speed ofembryo generation descends slower..when treating duration is same 7679-2、7680'sembryo number reduced accelerative by increase EMS concentration, and the more the concentration is the speed of embryo generation descends faster..
3. Every material has its own medial lethal concentration(LC5o): 7679-1 producing embryoes was about half of the CK when EMS concentration was 0.04% and the duration of EMS treatment was 10min; 7679-2 producing embryoes was about half of the CK when EMS concentration was 0.08% and the duration of EMS treatment was 10-20min;7680 producing embryoes was about half of the CK when EMS concentration was 0.04% and the duration of EMS treatment was 20min.
4. The phenotypic variation has get from DH plants' by EMS mutagenesis. Variation in economic character : height decreases, silique becomes shorter , branching number increases. Flower organ morphological variations including : pink petal, white peta; got more than six stamens, more than fore petalsl, and stigma exsertion. Qualitive variation:got a DH plant whose oil content achieved 43.81% (Donor adult plant index of oil 38.12-40.93%), a DH plant whose oleic acid content was 68.64% (Donor adult plant oleic acid content 62.93-65.82%).
5. EMS treatment influence the chromosome double rate of microspore-derived plants. In 7679-2's regeneration group, CK chromosome double rate was 54.0% by treating with colchicines, regeneration groups which treated by EMS before colchicines treating that chromosome double rate were 50.0-92.4%, were mostly higher than CK. In 7680's regeneration group, CK chromosome double rate was 96.7% by treating with colchicines; regeneration group which treated by EMS before colchicines treating that chromosome double rate were 85.7-98.0%. In a certain amount of concentration, EMS mutagenesis treatment approved the double of regeneration plant chromosome.
引文
1.陈军,陈正华,刘澄青,姚渝光,张丽华,关月兰.甘蓝型油菜游离小孢子培养和胚胎发生.遗传学报,1995,22(4):307-315
2.陈薇,李名扬,李宣源.离体茎尖诱变筛选油菜耐草酸变异体.西南农业学报,2001,14(2):31-33
3.陈文杰,谭小力,王竹云,李殿荣,周学秋.用傅立叶变换近红外光谱仪测定油菜种子品质指标的研究.陕西农业科学,2002(8):6-10
4.董遵,刘敬阳,马红梅,许才康,张建栋,孙华.离子束在油菜育种中的诱导效应初报[J].上海农业学报,2003,19(1):15-18
5.董遵,刘敬阳,牟健梅,张建栋.离子束处理油菜干种子的剂量效应.安徽农业科学,2005,33(6):949-950,956
6.杜连恩,于秀普,魏玉昌.EMS处理小麦合子诱变筛选高蛋白突变体[J].河北农业大学学报,1990,13(4):94-96
7.傅廷栋,杨光圣,涂金星,马朝芝.中国油菜生产的现状与展望.中国油脂,2003,28(1):11-13
8.顾佳清,张智奇,周音,奚银兴,张建军.EMS诱导水稻中花11突变体的筛选和鉴定.上海农业学报,2005,21(1):7-11
9.官春云.油菜小孢子培养和双倍体育种研究:Ⅰ供体植株和小孢子密度对小孢子培养的影响[J].作物学报,1995.21(6):665-670
10.官春云.油菜高油酸遗传育种研究进展.作物遗传,2006(1):1-8
11.郭会君,刘录祥,韩微波,赵世荣,李家才,赵林姝,王晶.高能混合粒子场辐照小麦的突变效应分析.中国农业科学,2008,41(3):654-660
12.郭丽娟,胡启德,康绍兰,张浩,黄梧芳.诱发玉米抗小斑病突变体的研究从玉米单倍体胚性细胞无性系筛选抗小斑病突变[J].遗传学报,1981,14(5)
13.何冬丽,杨光圣.甘蓝型油菜单倍体离体诱变及其效应的AFLP分子标记检测.中国油料作物学报,2004,26(2)
14.和江明,王敬乔,陈薇,李根泽,董云松,寸守铣.用EMS诱变和小孢子培养快速获得甘蓝型油菜高油酸种质材料的研究.西南农业学报,2003,16(2): 34-36
15.和江明,王敬乔,陈薇,李根泽,寸守铣.EMS对甘蓝型油菜离体小孢子胚胎发生能力的影响.西南农业学报,2004,17(6):690-693
16.胡婷婷.甘蓝型油菜与诸葛菜体细胞杂交研究.[硕士学位论文].武汉:华中农业大学图书馆,2006
17.姜淑慧,管荣展,董海滨,杜文明.播娘蒿与甘蓝型油菜的原生质体融合与植株再生.中国油料作物学报,2005,04:1-6
18.姜淑慧,管荣展,唐三元,忻如颖,张红生,赵立茜,潘琴燕.甘蓝型油菜与蔊菜的原生质体融合与植株再生.遗传,2007,29(6)
19.李栒,官春云,陈社元,王国槐.油菜小孢子培养和双单倍体育种研究Ⅱ影响甘蓝型油菜和芥菜型油菜种间杂种胚胎产量的因素.作物学报,2003,29(5):744-749
20.李再云.生物技术的油菜种质创新中的应用.生命科学研究,2006,10(1):205-214
21.刘泽,赵仁渠.空间条件对油菜诱变效果的研究-突变类型的观察与筛选.中国油料作物学报,2000a,22(2):6-8
22.刘泽,周永明.甘蓝型油菜离体小孢子胚胎发生能力的遗传分析.作物学报,2000b,26(1):104-109
23.刘良宏,石淑稳,吴江生,陈玉霞,周永明.油菜诱变和离体草酸筛选抗菌核病材料.中国油料作物学报,2003,25(1):5-8
24.刘志斋,蔡一林,王久光,孙海燕,王国强,杨春蓉,徐德林.EMS处理对玉米自交配合力的影响.玉米科学,2007,15(1):29-32,40
25.刘进平,郑成木.化学诱变结合离体选择选育胡椒抗瘟病无性系.热带作物学报,2006,27(1):22-27
26.刘勇,刘红雨,曾正宜.利用小孢子培养技术筛选油菜抗菌核病材料.西南农业大学,1997,1(10):108-112
27.刘勇,刘红雨,曾正宜.甘蓝型油菜小孢子胚状体的有道和培养.西南农业学报,1997-2-6
28.刘志文,刘雪平,傅廷栋,涂金星.甘蓝型油菜小孢子培养的胚诱导和加倍效率的研究.华中农业大学学报,2005,24(4):339-342
29.刘垂玎.微生物诱变的统计学研究.遗传学报1981,8(4):302-309
30.刘旻.白菜和诸葛菜杂交中的基因组加倍、染色体消除及偏白菜型杂种的产生.[硕士学位论文].武汉:华中农业大学图书馆,2006
31.罗鹏,傅华龙,蓝泽蘧,周颂东,周洪芳,罗晴.甘蓝型油菜与紫罗兰属间杂交的植物遗传学研究.植物学报,2003-45-4
32.牛应泽,刘玉贞,汪良中,袁有喜,李首成,范巧佳.人工合成甘蓝型油菜游离小孢子培养及其植株再生研究初报.四川农业大学学报,1999,17(2):167-171
33.牛应泽,刘玉贞,郭世星.提高甘蓝型油菜游离小孢子产胚率的研究.西南农业学报,2002,15(1):24-28
34.饶勇,徐涵,毛堂芬,李超,陈静,肖华贵.单倍体育种技术在油菜育种材料创新上的应用研究Ⅰ甘蓝型油菜游离小孢子胚状体的诱导发生.种子,2003,127:66-67
35.谌利,李加纳,王瑞.~(60)Co-γ对不同遗传背景甘蓝型黄籽油菜种子发芽力的影响.中国油料作物学报,1997,19(4):7-10
36.石淑稳,刘后利.甘蓝型油菜及种间和属间杂种小孢子胚状体的诱导[J].华中农业大学学报,1993,12(6):544-550
37.石淑稳,李再云,刘后利.甘蓝型油菜与诸葛菜的杂种小孢子胚的发生和小苗的形成.中国油料,1994,16(1):63-64
38.石淑稳,吴江生,刘后利.离体诱发甘蓝型油菜长角果和矮秆突变体.核农学报,1995,9(4):252-253
39.石淑稳,周永明,魏泽兰,吴江生,刘后利.甘蓝型油菜矮秆突变体DS-1和DS-2.中国种业,1997,3
40.石淑稳,周永明,吴江生,刘后利.甘蓝型油菜小孢子培养、染色体加倍、试管苗继代越夏和田间移栽配套技术的研究及其在油菜育种中的应用.中国农业通报,2001,17(2):57-60
41.石淑稳,周永明,吴江生.秋水仙碱处理油菜离体小孢子的染色体加倍效应.华中农业大学学报,2002,21(4):329-333
42.宋来强,贺兴文,邹晓芬,程春明,张建模,熊任香.甘蓝型油菜小孢子胚状体诱导的主要影响因素研究.江西农业学报,1998,10(2):66-69
43.王存喜.中华猕猴桃耐盐突变体筛选.核农学报,1990,4(4):206-210
44.王汉中,K Falk,,Downey R K.甘蓝型油菜Pol-CMS不同保持系和恢复系的游离小孢子培养和植株再生[A].中国作物学会油料作物专业委会.中国油料作物科学技术新进展[A].北京,中国农业科技出版社,1996
45.王培英,王连铮.EMS诱发大豆脂肪酸组成优良突变的研究[J].核农学报,1993,7(2):81-87
46.王亦菲,陆瑞菊,孙月芳,周润梅,刘成洪,黄剑华.大田油菜游离小孢子培养高频胚状体诱导及植株再生.中国农业通报,2002,18(1):20-24
47.魏良明,姜鸿勋,胡学安,赵发欣.植物诱变新技术及其在玉米育种上的应用[J].玉米科学,2002,8(1):19-20
48.吴江生,石淑稳,周永明,刘后利.甘蓝型双低油菜品种华双3号的选育和研究.华中农业大学学报,1999,1
49.邬贤梦,官春云,李栒.油菜脂肪酸品质改良的研究进展.作物研究,2003,17(3)
50.徐冠仁.植物诱变育种学[C].北京:中国农业出版社,1992,63-74
53.余风群.甘蓝型油菜未成熟小孢子堵养体系的研究[D].[博士学位论文].武汉:华中农业大学图书馆,1994
51.余凤群,刘后利.供体材料和培养基成分对甘蓝型油菜小孢子胚状体产生的影响.华中农业大学学报,1995.14(4):327-331
52.余凤群,刘后利.提高甘蓝型油菜小孢子胚状体成苗率的某些培养因素研究.作物学报,1997,23(2):165-168
54.张晓伟,千田泰义,栗根义,河野满,西口郁夫.一个苔用油菜品种的游离小孢子培养.全国蔬菜遗传育种学术讨论会,2002,82-87
55.张凤兰,高田.甘蓝型油菜小孢子胚胎发生能力的遗传分析.华北农学报,2001,16(1):27-32
56.张宏军.EMS处理对甘蓝型油菜的影响及Fad2基因的克隆和序列分析.[硕士学位论文].长沙,湖南农业大学,2007
57.张永泰,李爱民,陆莉,陈柳,惠飞虎,王幼平.通过甘蓝型油菜和白芥属间杂交后代的小孢子培养获得二倍体异附加系.作物学报,2006,32(11):1764-1766
58.钟维瑾,方光华,唐克轩,张智奇,俞妙娟.甘蓝型油菜游离小孢子培养诱导 胚状体再生植株.上海农业学报,1990,6(4):11-16
59.周嘉平,周俭民,梁思信,黄河,庞龙,校迎宪.烟草抗黑胫病突变体的细胞筛选.遗传学报,1990,17(3):180-188
60.朱保葛,路子显,耿玉轩,邓向东,谷爱秋.烷化剂EMS诱发花生性状变异的效果及高产突变系的选育.中国农业科学,1997,30(6):87-89
61.祝丽英,池书敏,刘志增,孟义江,陈景堂.EMS玉米花粉诱变的M_1代生物学效应.玉米科学,2002,10(1):33-35
62.Ahmad L.In vitro selection of primary embryos derived from UV-treated microspores of rapid cycling Brassica napus for herbicide tolerance.Cruciferae Newsletter,1991,(14-15):86-87
63.Ahamd L,Day J P,Macdonald M V and Ingram D S.Haploid culture and UV mutagenesis in rapid-cycling Brassica napus for the generation of resistance to chlorsulfuron and Alternaria bras sicicola.Annals of Botany,1991,67:521-525
64.Barro F,Fernandez Eacober J,Dela MV.Doubled haploid lines of Brassica carinata with modified erucic acid content through mutagenesis by EMS treatment of isolated microspores.Plant Breeding,2001,120:262-264.
65.Barro F,Fernandez Escobar J,Dela MV,Martin A.Modification of glucosinolate and erucic acid contents in doubled haploid lines of Brassica carinata by UV treatment of isolated microspore.Euphytica,2003,129(1):1-6
66.Bhagwat B,Duncan E J.Mutation breeding of Banana cv.Highgate(Musa spp,AAA Group)for tolerence to Fusorium oxysporum f.sp.Cubense using chemical mutagens.Scientia Horticulture,1998,73:11-22
67.Chuong P V,Deslauriers C,Kott L S,Beversdsorf WD Effects of donor genotype and bud sampling on microspore culture of Brassica napus.Can J Bot,1988,66:1653-1657
68.Cloutier S,Cappadocia M,Landry B S.Study of microspore-culture responsiveness in oilseed rape(brassica napus L) by comparative mapping of a F_2 population and two microspore-derived populations.Theor Appl Genet,1995,91:841-848
69.Dias J S,Correia MC.Effect of medium renovation and incubation temperature regimes on tronchuda cabbage microspore culture embryogenesis.Sci.Hortic.2002, 93
70. Fletcher R, Coventry J, Kott L S. Doubled haploid technology for winter and winter Brassica napus. Guelph, Ontario, Canada: OAC Publication , University of Guelph,1998
71. Gland A, Lichter R, Schweiger HG. Genetic and exogenous factors affecting embryogenesis in isolated microspore cultures of Brassica napus L. J Plant Physiol,1988, 132:613-617
72. Guha S, Maheshwari S C. In vitro production of embryoes from anthers of Datura.Nature, 1964-204-497
73. Hanson N J P, Anderson S B. In vitro chromosome doubling potential of colchicine,oryzalin, trifluralin and APM in Brassica napus microspore culture. Euphytica, 1996,88: 159-164
74. Iqbal M C M, Mollers C, Robbelen G. Increased embryogenesis after colchicine treatment of microspore cultures of Brassica napus L. Journal of Plant Physiology,1994, 143: 222-226
75. Kott LS. Production of mutants using the rapeseed doubled aploid system [ A ]. FAO/IAEA. Induced Mutation and olecular techniques for crop improvement. IAEA/FAO proceedings of an international symposiumon the use of induced mutations and molecular techniques for crop improvement. Vienne, Austria: IAEA 1996. 505-515
76. Lebkowski J H. Miller and M P Calos Determination of DNA sequence changes induced by ethyl methane sulfonate in human cell,using a shuttle vector system. Mol.Cell. Biol, 1986, 6: 1838-1842
77. Leonardo, Fernandez Martinez J M. De Haro A Intraspecific breeding for reduced glucosinolate content in Ethipian mustand. Euphytica, 1999, 106(2): 125-130
78. Lichter R. Induction of haploid plants from isolated pollen of Brassica napus .Pflanzen physiol, 1982, 105: 427-434
79. Lionneton E, Beuret W, Delaitre C, Ochatt S, Rancillac M. Improved microspore culture and doubled-haploid plant regeneration in the brown condiment mustard(Brassica juncea). Plant Cell Rep, 2001, 20: 126-130
80. Mathiae R. An Imporoved in vitro culture procedure for Embryoids Derived from isolsted microspores of rape {Brassica napus I), 1992, 100: 320-322
81. Neuffer M C. Paraffin oil technique for treating mature corn pollen whit mutagens [J].Maydic, 1978,22:21-28
82. Philip Raney J J, Gerhard FW Rakow, Todd V Olson. Indentification of Brassica napus germplasm with seed oil low in saturated fat. Proceedings of the 10th internafional rapeseed congress, Canberra Australia, 1999
83. Rucker B, Robbelen G. Development of high oleic acid rapeseed[A]. Proc 9th Int Rapeseed Congress ( GCIRC), Cambridge, 1995. 389-391
84. Sacristan MD. Resistance responses to phoma lingam of plants regenerated from selected cell and embryogenic cultures of haploid Brassica napus. TAG, 1982, 61:193-200
85. Somers D J, Friesen K R D, Rakow G. Identification of molecular markers associated with linoleic acid desatura. tion in Brassica napus. Theor Appl genet, 1998, 96:897-903
86. Swanson E B, Coumans MP, Bown GL, Patel J D, Beversdorf W D. The characterization of herbicide toleant plants in Brassica napus L. after in vitro selection of microspores and potoplasts. Plant Cell Reports,1988, 7: 83-87
87. Teresa CT, Laurencja S, Jan K. An in vitro mutagenesis-selection system for Brassica napus L. [A]Proceedings of the 10th international rapeseed congress. Canberra,Australia, 1999
88. Thuling N, Depittayananv. EMS induction of esly floweing mutants in sping ape {Brassica napus). Plant Beeding, 1992, 108: 177-184
89. Tonnemake K A, Auld DL, Thill DC, Mallory Smith CA.and Erickson DA..Development of sulfonylurea-resistant Rapeseed using chemical mutagenesis. Cop Science, 1992,32: 1387-1391
90. Wang H Z. Application of microspore culture technology in the breeding of rapeseed hybrids[A]. Wratten N, Salisbury P A. Proc 10th Intern Rap Cong. Canberra,Australia, 1999, 264-269
91. Yasui T, Sasaki T, Matsuki J,Yamamori M. Waxy endosperm mutants of bread wheat(Triticum aestivum L)and their starch properties. Breeding Science, 1997, 47:161-163
92. Zaki M, Dickinson H. Modification of cell development in vitro:the effect of colchicine on anther and isolated microspore culture in Brassica napus[J]. Plant Cell,Tissue and Organ Culture, 1995, 40: 255-270
93. Zhang F L, Takahata Y. Inheritance of microspore embryogenic ability in Brassica crops. Theor Appl Genet, 2001, 103: 254-256
2.陈薇,李名扬,李宣源.离体茎尖诱变筛选油菜耐草酸变异体.西南农业学报,2001,14(2):31-33
3.陈文杰,谭小力,王竹云,李殿荣,周学秋.用傅立叶变换近红外光谱仪测定油菜种子品质指标的研究.陕西农业科学,2002(8):6-10
4.董遵,刘敬阳,马红梅,许才康,张建栋,孙华.离子束在油菜育种中的诱导效应初报[J].上海农业学报,2003,19(1):15-18
5.董遵,刘敬阳,牟健梅,张建栋.离子束处理油菜干种子的剂量效应.安徽农业科学,2005,33(6):949-950,956
6.杜连恩,于秀普,魏玉昌.EMS处理小麦合子诱变筛选高蛋白突变体[J].河北农业大学学报,1990,13(4):94-96
7.傅廷栋,杨光圣,涂金星,马朝芝.中国油菜生产的现状与展望.中国油脂,2003,28(1):11-13
8.顾佳清,张智奇,周音,奚银兴,张建军.EMS诱导水稻中花11突变体的筛选和鉴定.上海农业学报,2005,21(1):7-11
9.官春云.油菜小孢子培养和双倍体育种研究:Ⅰ供体植株和小孢子密度对小孢子培养的影响[J].作物学报,1995.21(6):665-670
10.官春云.油菜高油酸遗传育种研究进展.作物遗传,2006(1):1-8
11.郭会君,刘录祥,韩微波,赵世荣,李家才,赵林姝,王晶.高能混合粒子场辐照小麦的突变效应分析.中国农业科学,2008,41(3):654-660
12.郭丽娟,胡启德,康绍兰,张浩,黄梧芳.诱发玉米抗小斑病突变体的研究从玉米单倍体胚性细胞无性系筛选抗小斑病突变[J].遗传学报,1981,14(5)
13.何冬丽,杨光圣.甘蓝型油菜单倍体离体诱变及其效应的AFLP分子标记检测.中国油料作物学报,2004,26(2)
14.和江明,王敬乔,陈薇,李根泽,董云松,寸守铣.用EMS诱变和小孢子培养快速获得甘蓝型油菜高油酸种质材料的研究.西南农业学报,2003,16(2): 34-36
15.和江明,王敬乔,陈薇,李根泽,寸守铣.EMS对甘蓝型油菜离体小孢子胚胎发生能力的影响.西南农业学报,2004,17(6):690-693
16.胡婷婷.甘蓝型油菜与诸葛菜体细胞杂交研究.[硕士学位论文].武汉:华中农业大学图书馆,2006
17.姜淑慧,管荣展,董海滨,杜文明.播娘蒿与甘蓝型油菜的原生质体融合与植株再生.中国油料作物学报,2005,04:1-6
18.姜淑慧,管荣展,唐三元,忻如颖,张红生,赵立茜,潘琴燕.甘蓝型油菜与蔊菜的原生质体融合与植株再生.遗传,2007,29(6)
19.李栒,官春云,陈社元,王国槐.油菜小孢子培养和双单倍体育种研究Ⅱ影响甘蓝型油菜和芥菜型油菜种间杂种胚胎产量的因素.作物学报,2003,29(5):744-749
20.李再云.生物技术的油菜种质创新中的应用.生命科学研究,2006,10(1):205-214
21.刘泽,赵仁渠.空间条件对油菜诱变效果的研究-突变类型的观察与筛选.中国油料作物学报,2000a,22(2):6-8
22.刘泽,周永明.甘蓝型油菜离体小孢子胚胎发生能力的遗传分析.作物学报,2000b,26(1):104-109
23.刘良宏,石淑稳,吴江生,陈玉霞,周永明.油菜诱变和离体草酸筛选抗菌核病材料.中国油料作物学报,2003,25(1):5-8
24.刘志斋,蔡一林,王久光,孙海燕,王国强,杨春蓉,徐德林.EMS处理对玉米自交配合力的影响.玉米科学,2007,15(1):29-32,40
25.刘进平,郑成木.化学诱变结合离体选择选育胡椒抗瘟病无性系.热带作物学报,2006,27(1):22-27
26.刘勇,刘红雨,曾正宜.利用小孢子培养技术筛选油菜抗菌核病材料.西南农业大学,1997,1(10):108-112
27.刘勇,刘红雨,曾正宜.甘蓝型油菜小孢子胚状体的有道和培养.西南农业学报,1997-2-6
28.刘志文,刘雪平,傅廷栋,涂金星.甘蓝型油菜小孢子培养的胚诱导和加倍效率的研究.华中农业大学学报,2005,24(4):339-342
29.刘垂玎.微生物诱变的统计学研究.遗传学报1981,8(4):302-309
30.刘旻.白菜和诸葛菜杂交中的基因组加倍、染色体消除及偏白菜型杂种的产生.[硕士学位论文].武汉:华中农业大学图书馆,2006
31.罗鹏,傅华龙,蓝泽蘧,周颂东,周洪芳,罗晴.甘蓝型油菜与紫罗兰属间杂交的植物遗传学研究.植物学报,2003-45-4
32.牛应泽,刘玉贞,汪良中,袁有喜,李首成,范巧佳.人工合成甘蓝型油菜游离小孢子培养及其植株再生研究初报.四川农业大学学报,1999,17(2):167-171
33.牛应泽,刘玉贞,郭世星.提高甘蓝型油菜游离小孢子产胚率的研究.西南农业学报,2002,15(1):24-28
34.饶勇,徐涵,毛堂芬,李超,陈静,肖华贵.单倍体育种技术在油菜育种材料创新上的应用研究Ⅰ甘蓝型油菜游离小孢子胚状体的诱导发生.种子,2003,127:66-67
35.谌利,李加纳,王瑞.~(60)Co-γ对不同遗传背景甘蓝型黄籽油菜种子发芽力的影响.中国油料作物学报,1997,19(4):7-10
36.石淑稳,刘后利.甘蓝型油菜及种间和属间杂种小孢子胚状体的诱导[J].华中农业大学学报,1993,12(6):544-550
37.石淑稳,李再云,刘后利.甘蓝型油菜与诸葛菜的杂种小孢子胚的发生和小苗的形成.中国油料,1994,16(1):63-64
38.石淑稳,吴江生,刘后利.离体诱发甘蓝型油菜长角果和矮秆突变体.核农学报,1995,9(4):252-253
39.石淑稳,周永明,魏泽兰,吴江生,刘后利.甘蓝型油菜矮秆突变体DS-1和DS-2.中国种业,1997,3
40.石淑稳,周永明,吴江生,刘后利.甘蓝型油菜小孢子培养、染色体加倍、试管苗继代越夏和田间移栽配套技术的研究及其在油菜育种中的应用.中国农业通报,2001,17(2):57-60
41.石淑稳,周永明,吴江生.秋水仙碱处理油菜离体小孢子的染色体加倍效应.华中农业大学学报,2002,21(4):329-333
42.宋来强,贺兴文,邹晓芬,程春明,张建模,熊任香.甘蓝型油菜小孢子胚状体诱导的主要影响因素研究.江西农业学报,1998,10(2):66-69
43.王存喜.中华猕猴桃耐盐突变体筛选.核农学报,1990,4(4):206-210
44.王汉中,K Falk,,Downey R K.甘蓝型油菜Pol-CMS不同保持系和恢复系的游离小孢子培养和植株再生[A].中国作物学会油料作物专业委会.中国油料作物科学技术新进展[A].北京,中国农业科技出版社,1996
45.王培英,王连铮.EMS诱发大豆脂肪酸组成优良突变的研究[J].核农学报,1993,7(2):81-87
46.王亦菲,陆瑞菊,孙月芳,周润梅,刘成洪,黄剑华.大田油菜游离小孢子培养高频胚状体诱导及植株再生.中国农业通报,2002,18(1):20-24
47.魏良明,姜鸿勋,胡学安,赵发欣.植物诱变新技术及其在玉米育种上的应用[J].玉米科学,2002,8(1):19-20
48.吴江生,石淑稳,周永明,刘后利.甘蓝型双低油菜品种华双3号的选育和研究.华中农业大学学报,1999,1
49.邬贤梦,官春云,李栒.油菜脂肪酸品质改良的研究进展.作物研究,2003,17(3)
50.徐冠仁.植物诱变育种学[C].北京:中国农业出版社,1992,63-74
53.余风群.甘蓝型油菜未成熟小孢子堵养体系的研究[D].[博士学位论文].武汉:华中农业大学图书馆,1994
51.余凤群,刘后利.供体材料和培养基成分对甘蓝型油菜小孢子胚状体产生的影响.华中农业大学学报,1995.14(4):327-331
52.余凤群,刘后利.提高甘蓝型油菜小孢子胚状体成苗率的某些培养因素研究.作物学报,1997,23(2):165-168
54.张晓伟,千田泰义,栗根义,河野满,西口郁夫.一个苔用油菜品种的游离小孢子培养.全国蔬菜遗传育种学术讨论会,2002,82-87
55.张凤兰,高田.甘蓝型油菜小孢子胚胎发生能力的遗传分析.华北农学报,2001,16(1):27-32
56.张宏军.EMS处理对甘蓝型油菜的影响及Fad2基因的克隆和序列分析.[硕士学位论文].长沙,湖南农业大学,2007
57.张永泰,李爱民,陆莉,陈柳,惠飞虎,王幼平.通过甘蓝型油菜和白芥属间杂交后代的小孢子培养获得二倍体异附加系.作物学报,2006,32(11):1764-1766
58.钟维瑾,方光华,唐克轩,张智奇,俞妙娟.甘蓝型油菜游离小孢子培养诱导 胚状体再生植株.上海农业学报,1990,6(4):11-16
59.周嘉平,周俭民,梁思信,黄河,庞龙,校迎宪.烟草抗黑胫病突变体的细胞筛选.遗传学报,1990,17(3):180-188
60.朱保葛,路子显,耿玉轩,邓向东,谷爱秋.烷化剂EMS诱发花生性状变异的效果及高产突变系的选育.中国农业科学,1997,30(6):87-89
61.祝丽英,池书敏,刘志增,孟义江,陈景堂.EMS玉米花粉诱变的M_1代生物学效应.玉米科学,2002,10(1):33-35
62.Ahmad L.In vitro selection of primary embryos derived from UV-treated microspores of rapid cycling Brassica napus for herbicide tolerance.Cruciferae Newsletter,1991,(14-15):86-87
63.Ahamd L,Day J P,Macdonald M V and Ingram D S.Haploid culture and UV mutagenesis in rapid-cycling Brassica napus for the generation of resistance to chlorsulfuron and Alternaria bras sicicola.Annals of Botany,1991,67:521-525
64.Barro F,Fernandez Eacober J,Dela MV.Doubled haploid lines of Brassica carinata with modified erucic acid content through mutagenesis by EMS treatment of isolated microspores.Plant Breeding,2001,120:262-264.
65.Barro F,Fernandez Escobar J,Dela MV,Martin A.Modification of glucosinolate and erucic acid contents in doubled haploid lines of Brassica carinata by UV treatment of isolated microspore.Euphytica,2003,129(1):1-6
66.Bhagwat B,Duncan E J.Mutation breeding of Banana cv.Highgate(Musa spp,AAA Group)for tolerence to Fusorium oxysporum f.sp.Cubense using chemical mutagens.Scientia Horticulture,1998,73:11-22
67.Chuong P V,Deslauriers C,Kott L S,Beversdsorf WD Effects of donor genotype and bud sampling on microspore culture of Brassica napus.Can J Bot,1988,66:1653-1657
68.Cloutier S,Cappadocia M,Landry B S.Study of microspore-culture responsiveness in oilseed rape(brassica napus L) by comparative mapping of a F_2 population and two microspore-derived populations.Theor Appl Genet,1995,91:841-848
69.Dias J S,Correia MC.Effect of medium renovation and incubation temperature regimes on tronchuda cabbage microspore culture embryogenesis.Sci.Hortic.2002, 93
70. Fletcher R, Coventry J, Kott L S. Doubled haploid technology for winter and winter Brassica napus. Guelph, Ontario, Canada: OAC Publication , University of Guelph,1998
71. Gland A, Lichter R, Schweiger HG. Genetic and exogenous factors affecting embryogenesis in isolated microspore cultures of Brassica napus L. J Plant Physiol,1988, 132:613-617
72. Guha S, Maheshwari S C. In vitro production of embryoes from anthers of Datura.Nature, 1964-204-497
73. Hanson N J P, Anderson S B. In vitro chromosome doubling potential of colchicine,oryzalin, trifluralin and APM in Brassica napus microspore culture. Euphytica, 1996,88: 159-164
74. Iqbal M C M, Mollers C, Robbelen G. Increased embryogenesis after colchicine treatment of microspore cultures of Brassica napus L. Journal of Plant Physiology,1994, 143: 222-226
75. Kott LS. Production of mutants using the rapeseed doubled aploid system [ A ]. FAO/IAEA. Induced Mutation and olecular techniques for crop improvement. IAEA/FAO proceedings of an international symposiumon the use of induced mutations and molecular techniques for crop improvement. Vienne, Austria: IAEA 1996. 505-515
76. Lebkowski J H. Miller and M P Calos Determination of DNA sequence changes induced by ethyl methane sulfonate in human cell,using a shuttle vector system. Mol.Cell. Biol, 1986, 6: 1838-1842
77. Leonardo, Fernandez Martinez J M. De Haro A Intraspecific breeding for reduced glucosinolate content in Ethipian mustand. Euphytica, 1999, 106(2): 125-130
78. Lichter R. Induction of haploid plants from isolated pollen of Brassica napus .Pflanzen physiol, 1982, 105: 427-434
79. Lionneton E, Beuret W, Delaitre C, Ochatt S, Rancillac M. Improved microspore culture and doubled-haploid plant regeneration in the brown condiment mustard(Brassica juncea). Plant Cell Rep, 2001, 20: 126-130
80. Mathiae R. An Imporoved in vitro culture procedure for Embryoids Derived from isolsted microspores of rape {Brassica napus I), 1992, 100: 320-322
81. Neuffer M C. Paraffin oil technique for treating mature corn pollen whit mutagens [J].Maydic, 1978,22:21-28
82. Philip Raney J J, Gerhard FW Rakow, Todd V Olson. Indentification of Brassica napus germplasm with seed oil low in saturated fat. Proceedings of the 10th internafional rapeseed congress, Canberra Australia, 1999
83. Rucker B, Robbelen G. Development of high oleic acid rapeseed[A]. Proc 9th Int Rapeseed Congress ( GCIRC), Cambridge, 1995. 389-391
84. Sacristan MD. Resistance responses to phoma lingam of plants regenerated from selected cell and embryogenic cultures of haploid Brassica napus. TAG, 1982, 61:193-200
85. Somers D J, Friesen K R D, Rakow G. Identification of molecular markers associated with linoleic acid desatura. tion in Brassica napus. Theor Appl genet, 1998, 96:897-903
86. Swanson E B, Coumans MP, Bown GL, Patel J D, Beversdorf W D. The characterization of herbicide toleant plants in Brassica napus L. after in vitro selection of microspores and potoplasts. Plant Cell Reports,1988, 7: 83-87
87. Teresa CT, Laurencja S, Jan K. An in vitro mutagenesis-selection system for Brassica napus L. [A]Proceedings of the 10th international rapeseed congress. Canberra,Australia, 1999
88. Thuling N, Depittayananv. EMS induction of esly floweing mutants in sping ape {Brassica napus). Plant Beeding, 1992, 108: 177-184
89. Tonnemake K A, Auld DL, Thill DC, Mallory Smith CA.and Erickson DA..Development of sulfonylurea-resistant Rapeseed using chemical mutagenesis. Cop Science, 1992,32: 1387-1391
90. Wang H Z. Application of microspore culture technology in the breeding of rapeseed hybrids[A]. Wratten N, Salisbury P A. Proc 10th Intern Rap Cong. Canberra,Australia, 1999, 264-269
91. Yasui T, Sasaki T, Matsuki J,Yamamori M. Waxy endosperm mutants of bread wheat(Triticum aestivum L)and their starch properties. Breeding Science, 1997, 47:161-163
92. Zaki M, Dickinson H. Modification of cell development in vitro:the effect of colchicine on anther and isolated microspore culture in Brassica napus[J]. Plant Cell,Tissue and Organ Culture, 1995, 40: 255-270
93. Zhang F L, Takahata Y. Inheritance of microspore embryogenic ability in Brassica crops. Theor Appl Genet, 2001, 103: 254-256
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