工业大麻雄性相关RAPD和SCAR标记的筛选与鉴定
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摘要
利用42条RAPD(Random Amplified Polymorphic DNA)随机扩增引物分析工业大麻品种"火麻一号"组成的雄性或雌性DNA池(DNA pools),结果显示,引物OPV-08扩增得到一条大小为869bp与工业大麻雄性相关的特异条带。根据测序结果,合成了两条SCAR(Sequence Characterized Amplified Region)标记引物,该SCAR标记不仅可以对工业大麻雌雄异株材料花期已知性别的雌雄植株进行准确鉴定,还可以对幼苗期未知性别的大麻雌雄植株进行鉴定;也可对雌雄同株材料可能出现的雄化进行早期鉴定。这不仅为工业大麻早期性别鉴定提供基础,且为减少雌雄同株材料的雄化提供支撑。
The male DNA pools or female DNA pools of "Huoma No.1" were analyzed respectively by 42 RAPD random amplified primers.The results showed that a male-specific DNA fragment with a length of 869 bp was amplified by OPV-08.Two SCAR primers were constructed according to the sequencing results.The primers of SCAR marker could identify the different genders of hemp not only in the known sex plants at flowering stage but also in the unknown sex seedlings at seedling stage.That could also be used for early identification of the appearance of male plant in monoecious industrial hemp.This provides a basis for early sex identification of industrial hemp,also provides support for reducing the appearance of male plant in monoecious industrial hemp.
The male DNA pools or female DNA pools of "Huoma No.1" were analyzed respectively by 42 RAPD random amplified primers.The results showed that a male-specific DNA fragment with a length of 869 bp was amplified by OPV-08.Two SCAR primers were constructed according to the sequencing results.The primers of SCAR marker could identify the different genders of hemp not only in the known sex plants at flowering stage but also in the unknown sex seedlings at seedling stage.That could also be used for early identification of the appearance of male plant in monoecious industrial hemp.This provides a basis for early sex identification of industrial hemp,also provides support for reducing the appearance of male plant in monoecious industrial hemp.
引文
[1]熊和平.麻类作物育种学.北京:中国农业科学技术出版社,2008:297.
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[4]Radu S,Robu T.Effects and efficiency of dietary hemp seed and flaxseed oils on the human metabolic function.Journal of Environmental Protection&Ecology,2014,15(1):326-331.
[5]吕佳淑,赵立宁,臧巩固,等.大麻性别相关AFLP分子标记筛选.湖南农业大学学报(自然科学版),2010,36(2):123-127.
[6]李仕金,辛培尧,周军,等.大麻性别研究进展.中国麻业科学,2008,30(2):110-113.
[7]Shao H,Clarke R C.Taxonomic studies of Cannabis in China.Journal of the International Hemp Association,1997,3(2):55-60.
[8]宋书娟,刘卉,邵宏.大麻性别连锁的特异DNA标记的初步研究.中国药物依赖性杂志,2001,10(3):182-184.
[9]李仕金,辛培尧,郭鸿彦,等.大麻雄性相关RAPD和SCAR标记的研究.广东农业科学,2012,39(24):151-154.
[10]陈美霞,祁建民,刘伟,等.麻类作物分子育种的研究现状与展望.福建农业学报,2012,27(7):780-786.
[11]郭丽,张海军,王明泽,等.大麻雄性基因连锁AFLP分子标记的筛选及鉴定.中国麻业科学,2015(1):5-8.
[12]张海军,郭丽,王明泽,等.大麻基因组DNA提取方法的比较与优化.中国麻业科学,2014,36(4):188-190.
[13]苏友波,朱颖,林春,等.大麻RAPD分子标记的引物筛选.中国麻业,2002,24(5):12-16.
[14]樊芳芳.大麻酚类化合物的全合成.郑州:郑州大学,2013.
[15]陈其本,余立惠,杨明,等.大麻栽培利用及发展对策.北京:电子科技大学出版社,1993:294-308.
[16]任朝兴.番木瓜(Carica papaya L.)遗传多样性的RAPD标记及雄性性别的RAPD和SCAR标记研究.重庆:西南大学,2007.
[17]Pauls,傅骏骅.应用RAPD分子标记识别苜蓿种群关系.作物杂志,1995(6):28-28.
[18]李向勇,陶波,李英慧,等.黑龙江省六个地点鸭跖草RAPD遗传多样性分析.作物杂志,2008(2):21-25.
[19]陈永胜,郝卫国,谭颖,等.RAPD标记蓖麻单雌性状的反应体系优化.作物杂志,2008(1):31-33.
[20]姜颖,孙宇峰,潘冬梅,等.工业大麻甲基磺酸乙酯(EMS)诱变体的筛选及RAPD分析.作物杂志,2017(6):50-54.
[21]关强,张月学,徐香玲,等.DNA分子标记的研究进展及几种新型分子标记技术.黑龙江农业科学,2008(1):102-104.
[22]邹继军,杨庆凯,陈受宜,等.大豆灰斑病抗病基因RAPD标记的分子特征及抗、感种质的SCAR标记鉴定.科学通报,1999,44(23):2544-2550.
[23]王剑锴.金线莲RAPD-SCAR标记的开发和不同种质遗传多样性评价.福州:福建农林大学,2016.
[24]闫华超,高岚,李桂兰.分子标记技术的发展及应用.生物学通报,2006,41(2):17-19.
[25]徐安毕,王文泉.几种分子标记方法相结合建立的新型分子标记方法.生物学通报,2007,42(1):26-28.
[26]Urasaki N,Tokumoto M,Tarora K,et al.A male and hermaphrodite specific RAPD marker for papaya(Carica papaya L.).Theoretical&Applied Genetics,2002,104(2/3):281-285.
[27]Xiong F,Zheng M J,Liu X R,et al.Development of SCAR markers based on RAPD and SRAP for rapid identification of pleurotus sajor-caju strain.Acta Agriculturae Universitatis Jiangxiensis,2010,32(3):601-607.
[28]Singh N,Pal A K,Meena B,et al.Development of ISSR-and RAPD-derived SCAR markers for identification of gladiolus germplasm.Journal of Horticultural Science&Biotechnology,2017,92(6):577-582.
[29]Zhou W,Wang Y,Zhang G,et al.Molecular sex identification in dioecious Hippophae rhamnoides L.via RAPD and SCAR markers.Molecules,2018,23(5):1048.
[30]Shao H,Song S J,Clarke R C.Female-associated DNApolymorphisms of hemp(Cannabis sativa L.).Journal of Industrial Hemp,2003,8(1):5-9.
[31]Mandolino G,Carboni A,Forapani S,et al.Identification of DNAmarkers linked to the male sex in dioecious hemp(Cannabis sativa L.).Theoretical&Applied Genetics,1999,98(1):86-92.
[32]陈其军,韩玉珍,傅永福,等.大麻性别的RAPD和SCAR分子标记.植物生理学报,2001,27(2):173-178.
[2]Allegret S,Bouloc P,Arnaud L,et al.Hemp:Industrial Production and Uses.Bouloc(Ed.),2013:4-26.
[3]Amaducci S,Scordia D,Liu F H,et al.Key cultivation techniques for hemp in Europe and China.Industrial Crops&Products,2014,68:2-16.
[4]Radu S,Robu T.Effects and efficiency of dietary hemp seed and flaxseed oils on the human metabolic function.Journal of Environmental Protection&Ecology,2014,15(1):326-331.
[5]吕佳淑,赵立宁,臧巩固,等.大麻性别相关AFLP分子标记筛选.湖南农业大学学报(自然科学版),2010,36(2):123-127.
[6]李仕金,辛培尧,周军,等.大麻性别研究进展.中国麻业科学,2008,30(2):110-113.
[7]Shao H,Clarke R C.Taxonomic studies of Cannabis in China.Journal of the International Hemp Association,1997,3(2):55-60.
[8]宋书娟,刘卉,邵宏.大麻性别连锁的特异DNA标记的初步研究.中国药物依赖性杂志,2001,10(3):182-184.
[9]李仕金,辛培尧,郭鸿彦,等.大麻雄性相关RAPD和SCAR标记的研究.广东农业科学,2012,39(24):151-154.
[10]陈美霞,祁建民,刘伟,等.麻类作物分子育种的研究现状与展望.福建农业学报,2012,27(7):780-786.
[11]郭丽,张海军,王明泽,等.大麻雄性基因连锁AFLP分子标记的筛选及鉴定.中国麻业科学,2015(1):5-8.
[12]张海军,郭丽,王明泽,等.大麻基因组DNA提取方法的比较与优化.中国麻业科学,2014,36(4):188-190.
[13]苏友波,朱颖,林春,等.大麻RAPD分子标记的引物筛选.中国麻业,2002,24(5):12-16.
[14]樊芳芳.大麻酚类化合物的全合成.郑州:郑州大学,2013.
[15]陈其本,余立惠,杨明,等.大麻栽培利用及发展对策.北京:电子科技大学出版社,1993:294-308.
[16]任朝兴.番木瓜(Carica papaya L.)遗传多样性的RAPD标记及雄性性别的RAPD和SCAR标记研究.重庆:西南大学,2007.
[17]Pauls,傅骏骅.应用RAPD分子标记识别苜蓿种群关系.作物杂志,1995(6):28-28.
[18]李向勇,陶波,李英慧,等.黑龙江省六个地点鸭跖草RAPD遗传多样性分析.作物杂志,2008(2):21-25.
[19]陈永胜,郝卫国,谭颖,等.RAPD标记蓖麻单雌性状的反应体系优化.作物杂志,2008(1):31-33.
[20]姜颖,孙宇峰,潘冬梅,等.工业大麻甲基磺酸乙酯(EMS)诱变体的筛选及RAPD分析.作物杂志,2017(6):50-54.
[21]关强,张月学,徐香玲,等.DNA分子标记的研究进展及几种新型分子标记技术.黑龙江农业科学,2008(1):102-104.
[22]邹继军,杨庆凯,陈受宜,等.大豆灰斑病抗病基因RAPD标记的分子特征及抗、感种质的SCAR标记鉴定.科学通报,1999,44(23):2544-2550.
[23]王剑锴.金线莲RAPD-SCAR标记的开发和不同种质遗传多样性评价.福州:福建农林大学,2016.
[24]闫华超,高岚,李桂兰.分子标记技术的发展及应用.生物学通报,2006,41(2):17-19.
[25]徐安毕,王文泉.几种分子标记方法相结合建立的新型分子标记方法.生物学通报,2007,42(1):26-28.
[26]Urasaki N,Tokumoto M,Tarora K,et al.A male and hermaphrodite specific RAPD marker for papaya(Carica papaya L.).Theoretical&Applied Genetics,2002,104(2/3):281-285.
[27]Xiong F,Zheng M J,Liu X R,et al.Development of SCAR markers based on RAPD and SRAP for rapid identification of pleurotus sajor-caju strain.Acta Agriculturae Universitatis Jiangxiensis,2010,32(3):601-607.
[28]Singh N,Pal A K,Meena B,et al.Development of ISSR-and RAPD-derived SCAR markers for identification of gladiolus germplasm.Journal of Horticultural Science&Biotechnology,2017,92(6):577-582.
[29]Zhou W,Wang Y,Zhang G,et al.Molecular sex identification in dioecious Hippophae rhamnoides L.via RAPD and SCAR markers.Molecules,2018,23(5):1048.
[30]Shao H,Song S J,Clarke R C.Female-associated DNApolymorphisms of hemp(Cannabis sativa L.).Journal of Industrial Hemp,2003,8(1):5-9.
[31]Mandolino G,Carboni A,Forapani S,et al.Identification of DNAmarkers linked to the male sex in dioecious hemp(Cannabis sativa L.).Theoretical&Applied Genetics,1999,98(1):86-92.
[32]陈其军,韩玉珍,傅永福,等.大麻性别的RAPD和SCAR分子标记.植物生理学报,2001,27(2):173-178.