象草不同辐射剂量诱变系表型及遗传变异研究
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摘要
以象草种茎为材料,利用10、20和30Gy剂量的60 Co-γ射线照射种茎,筛选适宜诱变剂量并研究其表型性状变异和分子水平上的遗传变异,为象草辐射诱变育种提供理论和技术基础。结果表明,辐射处理后的植株有矮化的趋势,其中分蘖数、茎节数和叶长对辐射最敏感,容易发生变异;在所有诱变系材料中,F30-39和F30-41聚为一类,与对照材料差异最大,是显著矮小化的植株。30Gy诱变系与未辐射的材料间的遗传相似系数在0.3793～0.9655,平均为0.8276,而差异位点数在4～66个,平均为19.3个,两项指标均大于10和20Gy诱变系群体。综合考虑辐射后植株存活率与遗传变异结果,选定30Gy为最适宜的象草种茎辐射诱变剂量。
In order to provide a theoretical basis for mutation-induced breeding of Pennisetum purpureum,stems of P.purpureum were irradiated with 60 Co-γrays at differing doses of 10,20 and 30 Gy.Then,we screened the resulting mutants and defined their phenotypic variation,and identified the genetic differences at a molecular level,using SSR.The irradiated plants showed a tendency to dwarfism.The number of tillers,stems and the length of leaves were the traits most sensitive to radiation and most easily mutated.Among all the mutants,two designated F30-39 and F30-41 were significantly shortened plants,and were determined by cluster analysis to belong to a group which had the greatest genetic difference from the control material.The genetic similarity coefficient between controls and"30 Gy"mutants ranged from 0.3793 to 0.9655,with the average being 0.8276,while the number of mutation sites ranged between 4 and 66,with an average of 19.3.Both indices were greater than those of"10 and 20 Gy"mutants.Considering the survival rate and genetic mutation frequency of irradiated plants,30 Gy was selected as the most suitable radiation dose level for mutagenesis.
In order to provide a theoretical basis for mutation-induced breeding of Pennisetum purpureum,stems of P.purpureum were irradiated with 60 Co-γrays at differing doses of 10,20 and 30 Gy.Then,we screened the resulting mutants and defined their phenotypic variation,and identified the genetic differences at a molecular level,using SSR.The irradiated plants showed a tendency to dwarfism.The number of tillers,stems and the length of leaves were the traits most sensitive to radiation and most easily mutated.Among all the mutants,two designated F30-39 and F30-41 were significantly shortened plants,and were determined by cluster analysis to belong to a group which had the greatest genetic difference from the control material.The genetic similarity coefficient between controls and"30 Gy"mutants ranged from 0.3793 to 0.9655,with the average being 0.8276,while the number of mutation sites ranged between 4 and 66,with an average of 19.3.Both indices were greater than those of"10 and 20 Gy"mutants.Considering the survival rate and genetic mutation frequency of irradiated plants,30 Gy was selected as the most suitable radiation dose level for mutagenesis.
引文
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[20] Wang W E,Bao M Z,Zhang J W.The effect of 60 Co-γirradiation on dry seeds of Zoysia japonica.Pratacultural Science,2009,26(5):155-160.王文恩,包满珠,张俊卫.60 Co-γ射线对日本结缕草干种子的辐射效应研究.草业科学,2009,26(5):155-160.
[21] Wang S N,Wang J H.Pennisetum purpureum Schum.‘Guiminyin’and its popularization and application.Fujian Journal of Animal Husbandry and Veterinary Medicine,2014,36(1):19-20.王舒宁,王均辉.桂闽引象草及其推广应用.福建畜牧兽医,2014,36(1):19-20.
[22] Tian H,Liu Y,Chen M X,et al.Preliminary studies on the application of Chinese Pennisetumas ornamental grass in landscape.Chinese Agricultural Science Bulletin,2012,28(1):307-310.田宏,刘洋,陈明新,等.浅析狼尾草作为观赏草在园林中的应用.中国农学通报,2012,28(1):307-310.
[23] Zeng J,Huang L K,Zhang X Q,et al.Study on phenotype mutation of Hemarthria compressa and dose selection of 60 Co-γirradiation.Acta Agrectir Sinica,2014,22(4):828-833.曾捷,黄琳凯,张新全,等.60 Co-γ辐射扁穗牛鞭草剂量筛选及表型变异研究.草地学报,2014,22(4):828-833.
[24] Zhang Y Q,Jia W L,Yang L L,et al.A study on character mutation of 60 Co-γray irradiation Festuca arundinacea.Acta Prataculturae Sinica,2005,14(4):65-71.张彦芹,贾炜珑,杨丽莉,等.60 Co-γ辐射高羊茅性状变异研究.草业学报,2005,14(4):65-71.
[25] Xuan J P,Guo A G,Liu J X.et al.Analysis on morphological variation of mutants of Cynodon dactylonindeed by radiation.Acta Prataculturae Sinica,2005,14(6):107-111.宣继萍,郭爱桂,刘建秀,等.狗牙根辐射诱变后代外部性状变异分析.草业学报,2005,14(6):107-111.
[26] Wang J,Wang S H,Lai Y,et al.Genetic diversity and population structure analysis by using SSR markers in barley.Acta Agriculturae Nucleatae Sinica,2014,28(2):177-185.王晋,王世红,赖勇,等.大麦SSR标记遗传多样性及群体遗传结构分析.核农学报,2014,28(2):177-185.
[27] Huang T,Ma X,Zhang X Q,et al.Comparation of SSR molecular markers analysis of annual ryegrass varieties in DUS testing.Scientia Agricultura Sinica,2015,48(2):381-389.黄婷,马啸,张新全,等.多花黑麦草DUS测定中SSR标记品种鉴定比较分析.中国农业科学,2015,48(2):381-389.
[28] You J H,Li S,He W Z,et al.A new cane variety Guitang No.22bred by 60 Co-γ-ray induced mutation.Acta Agriculturae Nucleatae Sinica,2006,20(2):95-98.游建华,李松,何为中,等.60 Co-γ辐射诱变育成甘蔗新品种桂糖22号.核农学报,2006,20(2):95-98.
[29] Liu T Z,Xie X C,Zhang J M.Mutagenic effect of 60 Co-γirradiation on turf characteristics of Paspalum vaginatum.Acta Prataculturae Sinica,2017,26(7):62-70.刘天增,谢新春,张巨明.海滨雀稗60 Co-γ辐射诱变突变体筛选.草业学报,2017,26(7):62-70.
[30] Sen A,Alikamanoglu S.Analysis of drought-tolerant sugar beet(Beta vulgaris L.)mutants induced with gamma radiation using SDS-PAGE and ISSR markers.Mutation Research,2012,738/739(1):38-44.
[31] Qin J Y,Shi H C,Ke Y P,et al.Research on genetic difference of maize mutants by irradiation based on phenotypic and SSR.Journal of Maize Sciences,2012,20(2):41-47.秦家友,石海春,柯永培,等.玉米辐射诱变系表型及SSR遗传差异研究.玉米科学,2012,20(2):41-47.
[2] Yao N,Lai Z Q,Yi X F,et al.Comparison of the production performance of four Pennisetum purpureum species.Guangxi Journal of Animal Husbandry&Veterinary Medicine,2015,(2):59-61.姚娜,赖志强,易显凤,等.四个象草品种生产性能的比较.广西畜牧兽医,2015,(2):59-61.
[3] Li M,Zi X J,Hou G Y,et al.Nutritional evaluation of five tropical gramineous forages.Acta Agrestia Sinica,2013,21(5):1028-1032.李茂,字学娟,侯冠彧,等.体外产气法评价5种热带禾本科牧草营养价值.草地学报,2013,21(5):1028-1032.
[4] Wang Q B.Selection and breeding of hardy plant of P.purpureumschumach by bud radiation.Guizhou Agricultural Sciences,2003,31(6):17-19.王庆斌.象草辐照芽变选育耐寒植株的初步研究.贵州农业科学,2003,31(6):17-19.
[5] Rong H,Xu A K,Xia T Y J,et al.Changes in fermentation quality of napiergrass harvested the first growth.Acta Agrestia Sinica,2009,17(4):537-539.荣辉,徐安凯,下条雅敬,等.初次刈割象草青贮发酵品质动态.草地学报,2009,17(4):537-539.
[6] Stricker J A,Smith D W.Economic development through biomass systems integration in central Florida.Golden,Colorado:National Renewable Energy Laboratory,1995.
[7] Jewell W J,Cummings R J,Richards B K.Methane fermentation of energy crops:Maximum conversion kinetics and in situ biogas purification.Biomass&Bioenergy,1993,5(314):261-278.
[8] Schank S C,Chynoweth D P,Turick C E,et al.Napiergrass genotypes and plant parts for biomass energy.Biomass&Bioenergy,1993,4(1):1-7.
[9] Woodard K R,Prine G M,Bachrein S.Solar energy recovery by elephantgrass,energycane,and elephantmillet canopies.Crop Science,1993,33(4):824-830.
[10] Wen X N,Jian Y Z,Xie X M.The comprehensive exploitation and utilization of Pennisetum purpureum.Pratacultural Science,2009,26(9):108-112.温晓娜,简有志,解新明.象草资源的综合开发利用.草业科学,2009,26(9):108-112.
[11] Xin J N,Xu W,Wang Z Y,et al.Application of Pennisetum alopecuroides as ornamental grass.Grassland and Turf,2013,(5):90-93.信金娜,徐威,王志英,等.狼尾草作为观赏草的应用.草原与草坪,2013,(5):90-93.
[12] Luo Z Z,Lin J R,Lin Z K,et al.Karyotype analysis of Pennisetum purpureum×P.americanumcv.Reyan No.4and P.purpureumcv.Guiminyin.Pratacultural Science,2016,33(9):1711-1717.罗宗志,林洁荣,林志魁,等.热研4号王草和桂闽引象草的核型分析.草业科学,2016,33(9):1711-1717.
[13] Chen Z T,He S L,Huang Y B.Research progress of Pennisetumrich.Acta Agrestia Sinica,2010,18(5):740-748.陈志彤,何水林,黄毅斌.狼尾草属牧草研究进展.草地学报,2010,18(5):740-748.
[14] Li Y H,Xie Z L,Xie X M.Characters and dynamics of modular biomass and its allocation ratio in five cultivars of elephant grass.Acta Prataculturae Sinica,2011,20(5):11-18.李有涵,谢昭良,解新明.5个象草品种的构件生物量特征及分配动态.草业学报,2011,20(5):11-18.
[15] Wang Z D.The progress of radiative mutation breeding in China.Journal of Isotopes,2005,18(3):183-185.王志东.我国辐射诱变育种的现状分析.同位素,2005,18(3):183-185.
[16] Ma H P,Zhao Y L,Yang G Y.Application of induced mutation technology for crop breeding.Hereditas,1998,20(4):48-50.马惠平,赵永亮,杨光宇.诱变技术在作物育种中的应用.遗传,1998,20(4):48-50.
[17] Han W B,Zhang Y X,Tang F L,et al.Research advances in pasture mutation breeding in China.Acta Agriculturae Nucleatae Sinica,2010,24(1):62-66.韩微波,张月学,唐凤兰,等.我国牧草诱变育种研究进展.核农学报,2010,24(1):62-66.
[18] Yun J F.Herbage and feed crop breeding.Beijing:China Agricultural Publishing House,2001.云锦凤.牧草及饲料作物育种学.北京:中国农业出版社,2001.
[19] Wang W E,Zhang J W,Bao M Z.60 Co-γray irradiation effect on germination and seedling growth of dry buchloe dactyloides seeds.Acta Agriculturae Nucleatae Sinica,2005,19(3):191-194.王文恩,张俊卫,包满珠.60 Co-γ辐射对野牛草干种子的刺激生长效应.核农学报,2005,19(3):191-194.
[20] Wang W E,Bao M Z,Zhang J W.The effect of 60 Co-γirradiation on dry seeds of Zoysia japonica.Pratacultural Science,2009,26(5):155-160.王文恩,包满珠,张俊卫.60 Co-γ射线对日本结缕草干种子的辐射效应研究.草业科学,2009,26(5):155-160.
[21] Wang S N,Wang J H.Pennisetum purpureum Schum.‘Guiminyin’and its popularization and application.Fujian Journal of Animal Husbandry and Veterinary Medicine,2014,36(1):19-20.王舒宁,王均辉.桂闽引象草及其推广应用.福建畜牧兽医,2014,36(1):19-20.
[22] Tian H,Liu Y,Chen M X,et al.Preliminary studies on the application of Chinese Pennisetumas ornamental grass in landscape.Chinese Agricultural Science Bulletin,2012,28(1):307-310.田宏,刘洋,陈明新,等.浅析狼尾草作为观赏草在园林中的应用.中国农学通报,2012,28(1):307-310.
[23] Zeng J,Huang L K,Zhang X Q,et al.Study on phenotype mutation of Hemarthria compressa and dose selection of 60 Co-γirradiation.Acta Agrectir Sinica,2014,22(4):828-833.曾捷,黄琳凯,张新全,等.60 Co-γ辐射扁穗牛鞭草剂量筛选及表型变异研究.草地学报,2014,22(4):828-833.
[24] Zhang Y Q,Jia W L,Yang L L,et al.A study on character mutation of 60 Co-γray irradiation Festuca arundinacea.Acta Prataculturae Sinica,2005,14(4):65-71.张彦芹,贾炜珑,杨丽莉,等.60 Co-γ辐射高羊茅性状变异研究.草业学报,2005,14(4):65-71.
[25] Xuan J P,Guo A G,Liu J X.et al.Analysis on morphological variation of mutants of Cynodon dactylonindeed by radiation.Acta Prataculturae Sinica,2005,14(6):107-111.宣继萍,郭爱桂,刘建秀,等.狗牙根辐射诱变后代外部性状变异分析.草业学报,2005,14(6):107-111.
[26] Wang J,Wang S H,Lai Y,et al.Genetic diversity and population structure analysis by using SSR markers in barley.Acta Agriculturae Nucleatae Sinica,2014,28(2):177-185.王晋,王世红,赖勇,等.大麦SSR标记遗传多样性及群体遗传结构分析.核农学报,2014,28(2):177-185.
[27] Huang T,Ma X,Zhang X Q,et al.Comparation of SSR molecular markers analysis of annual ryegrass varieties in DUS testing.Scientia Agricultura Sinica,2015,48(2):381-389.黄婷,马啸,张新全,等.多花黑麦草DUS测定中SSR标记品种鉴定比较分析.中国农业科学,2015,48(2):381-389.
[28] You J H,Li S,He W Z,et al.A new cane variety Guitang No.22bred by 60 Co-γ-ray induced mutation.Acta Agriculturae Nucleatae Sinica,2006,20(2):95-98.游建华,李松,何为中,等.60 Co-γ辐射诱变育成甘蔗新品种桂糖22号.核农学报,2006,20(2):95-98.
[29] Liu T Z,Xie X C,Zhang J M.Mutagenic effect of 60 Co-γirradiation on turf characteristics of Paspalum vaginatum.Acta Prataculturae Sinica,2017,26(7):62-70.刘天增,谢新春,张巨明.海滨雀稗60 Co-γ辐射诱变突变体筛选.草业学报,2017,26(7):62-70.
[30] Sen A,Alikamanoglu S.Analysis of drought-tolerant sugar beet(Beta vulgaris L.)mutants induced with gamma radiation using SDS-PAGE and ISSR markers.Mutation Research,2012,738/739(1):38-44.
[31] Qin J Y,Shi H C,Ke Y P,et al.Research on genetic difference of maize mutants by irradiation based on phenotypic and SSR.Journal of Maize Sciences,2012,20(2):41-47.秦家友,石海春,柯永培,等.玉米辐射诱变系表型及SSR遗传差异研究.玉米科学,2012,20(2):41-47.