毒素胁迫烟草抗黑胫病鉴定及抗性指标分析
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摘要
为建立一种高效准确的烟草黑胫病抗性鉴定方法,以4种具有不同抗性特性的烟草品种为材料,研究培养基附加不同浓度黑胫病病菌毒素及不同胁迫时间对烟草种子萌发和根系生长的影响。结果表明,烟草黑胫病病菌毒素胁迫对烟草种子的发芽率和根系生长均有抑制作用,抑制效果随胁迫浓度的增加而增强,且不同品种对毒素胁迫的耐受力不同,利用体积浓度为50%的毒素胁迫15 d时,抗病品种G28、中抗品种YN116、中感品种净叶黄、感病品种红花大金元的根系长进培养基中的株数比率分别为79.38%、70.74%、33.55%和13.36%。初步确定鉴别烟草种子抗性特性的最佳粗毒素胁迫浓度为50%(即含蛋白质13.53μg/m L,含糖19.64 mg/m L)、最佳鉴定时间为胁迫15 d,利用该方法可在短期内高效准确地鉴定出不同品种(种质)对烟草黑胫病的抗感水平,加快烟草抗黑胫病育种进程。
In order to establish an efficient and accurate method for identification of resistance to tobacco black shank, experiments were performed using 4 tobacco varieties with different resistance characteristics. The toxin was added to the medium to study the effects of different concentrations of toxin stress and different stress time on tobacco seed germination and root growth. The results showed that the toxin stress of black shank disease could inhibit the germination and root growth of tobacco seedlings, and the inhibitory effect increased with the increase of toxin concentration. Different varieties have different tolerance to toxin stress. When the toxin was used at a concentration of 50% for 15 days, the ratio of the number of plants in the root growth medium of G28, YN116,Jingyehuang and Hongda was 79.38%, 70.74%, 33.55% and 13.36%, respectively. It was preliminarily determined that the optimal crude toxin concentration for identifying the resistance level of different varieties(germplasm) to tobacco black shank was 50%(protein content 13.53 μg/mL, sugar content 19.64 mg/mL), and the optimal stress time was 15 days. This method can effectively and accurately identify the resistance level of different varieties(germplasm) to tobacco black shank in a short period of time, and accelerate the breeding process of tobacco resistance to black shank.
In order to establish an efficient and accurate method for identification of resistance to tobacco black shank, experiments were performed using 4 tobacco varieties with different resistance characteristics. The toxin was added to the medium to study the effects of different concentrations of toxin stress and different stress time on tobacco seed germination and root growth. The results showed that the toxin stress of black shank disease could inhibit the germination and root growth of tobacco seedlings, and the inhibitory effect increased with the increase of toxin concentration. Different varieties have different tolerance to toxin stress. When the toxin was used at a concentration of 50% for 15 days, the ratio of the number of plants in the root growth medium of G28, YN116,Jingyehuang and Hongda was 79.38%, 70.74%, 33.55% and 13.36%, respectively. It was preliminarily determined that the optimal crude toxin concentration for identifying the resistance level of different varieties(germplasm) to tobacco black shank was 50%(protein content 13.53 μg/mL, sugar content 19.64 mg/mL), and the optimal stress time was 15 days. This method can effectively and accurately identify the resistance level of different varieties(germplasm) to tobacco black shank in a short period of time, and accelerate the breeding process of tobacco resistance to black shank.
引文
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[13]王桥美,范静华,果志华,等.烟草黑胫病菌毒素对烟草防御性相关酶的诱导作用[J].云南农业大学学报,2011,26(1):20-25.WANG Q M,FAN J H,GUO Z H,et al.Induction effect of toxin extracts of phytophthora nicotianae on tobacco defense-related enzyme activities[J].Journal of Yunnan Agricultural University,2011,26(1):20-25.
[14]时焦,孙丽萍,孟坤,等.一种培养皿中筛选烟草抗赤星病种质材料的方法:中国,102876767 B[P].2013.01.16.SHI J,SUN L P,MENG K,et al.A method for screening tobacco anti-amazing disease germplasm material in culture dish:CN,102876767 B[P].2013.01.16.
[15]丁燕芳,平文丽,孙计平,等.烟草黑胫病抗性鉴定方法比较及其相关性分析[J].湖北农业科学,2017,56(18):3477-3480.DING Y F,PING W L,SUN J P,et al.Comparison of methods to identify resistance to tobacco black shank disease and their correlation analysis[J].Hubei Agricultural Sciences,2017,56(18):3477-3480.
[16]ZHANG Y S,GUO X,YAN X X,et al.Identification of stably expressed QTL for resistance to black shank disease in tobacco(Nicotiana tabacum L.)line Beinhart1000-1[J].The Crop Journal,2018,6(3):282-290.
[17]庞龙,黄河,周嘉平.烟草黑胫病菌毒素的分离纯化与性质的研究[J].科学通报,1986,31(18):1416-1416.PANG L,HUANG H,ZHOU J P.Isolation,purification and properties of toxin from tobacco black shank diseases[J].Science Bulletin,1986,31(18):1416-1420.
[18]高俊凤.植物生理学实验指导[M].北京:高等教育出版社,2006:140-148.GAO J F.Plant Physiology Experiment Guide[M].Beijing:Higher Education Press,2006:140-148.
[19]胡重怡,郑少清,陈兴江,等.烟草无菌苗培养前的种子消毒技术研究[J].中国烟草科学,2007,28(2):45-51.HU Z Y,ZHENG S Q,CHEN X J,et al.Methods for seed sterilization before aseptic tobacco seedling culture[J].Chinese Tobacco Science,2007,28(2):45-51.
[20]晏娟,黎定军.筛选烟草抗黑胫病细胞突变体的粗毒素浓度的初步研究[J].湖南农业科学,2011(5):66-68.YAN J,LI D J.A primary study on screening crude toxin concentration for cell mutants of tobacco against black shank[J].Hunan Agricultural Sciences,2011(5):66-68.
[21]高崇,吴国贺,李佰霖,等.延边烟区烟草菌核病病原菌鉴定及抗性种质筛选[J].中国烟草科学,2018,39(2):69-75.GAO C,WU G H,LI B L,et al.Identification of tobacco sclerotinia rot in Yanbian and screening for resistant germplasms[J].Chinese Tobacco Science,2018,39(2):69-75.
[22]叶协锋,张晓帆,郑宪滨,等.复合盐碱处理下烤烟品种发芽特性及耐盐性评价[J].中国烟草科学,2017,38(3):37-43.YE X F,ZHANG X F,ZHENG X B,et al.Germination characteristics of flue-cured tobacco varieties under mixed salt-alkali stresses and evaluations of their saline-alkali tolerance[J].Chinese Tobacco Science,2017,38(3):37-43.
[2]郭璇,闫杏杏,蒋彩虹,等.雪茄烟Beinhart1000-1对黑胫病0号生理小种的抗性遗传分析[J].中国烟草科学,2017,38(2):56-62.GUO X,YAN X X,JIANG C H,et al.Genetic analysis of beinhart1000-1 resistance to black shank in tobacco[J].Chinese Tobacco Science,2017,38(2):56-62.
[3]MCCORKLE K,LEWIS R,SHEW D.Resistance to phytophthora parasitica var.nicotianae in tobacco breeding Lines derived from variety beinhart 1000[J].Plant Disease,2013,97(2):252-258.
[4]马国胜,高智谋.烟草黑胫病菌培养性状的研究[J].中国农业科学,2007,40(3):512-517.MA G S,GAO Z M.Cultural characteristics of phytophthora nicotianae var.nicotianae,pathogen of tobacco blank shank[J].Scientia Agricultura Sinica,2007,40(3):512-517.
[5]黄丽华,陈廷俊,宋军.烟草黑胫病粗毒素高产培养条件的筛选[J].安徽农业科学,1999(4):368-370.HUANG L H,CHEN T J,SONG J.Selection of cultural conditions for higher toxin from the black shank of tobacco[J].Journal of Anhui Agricultural Sciences,1999(4):368-370.
[6]孙计平,李雪君,吴照辉,等.烟草黑胫病的研究进展[J].湖北农业科学,2011,50(16):3253-3256.SUN J P,LI X J,WU Z H,et al.Progress on tobacco black shank disease[J].Hubei Agricultural Sciences,2011,50(16):3253-3256.
[7]于海芹,刘勇,许美玲,等.新引进烟草种质资源的黑胫病抗性鉴定[J].基因组学与应用生物学,2015,34(5):1027-1033.YU H Q,LIU Y,XU M L,et al.Identification of resistance to black shank of new tobacco germplasm[J].Genomics&Applied Biology,2015,34(5):1027-1033.
[8]于海芹,焦芳婵,肖炳光,等.烟草种质资源苗期黑胫病抗性鉴定研究[J].中国农业科技导报,2008,10(4):70-75.YU H Q,JIAO F C,XIAO B G,et al.Studies on identification of resistance to black shank disease of tobacco gemplasm resources in seeding stage[J].Journal of Agricultural Science and Technology,2008,10(4):70-75.
[9]李梅云,刘勇,肖炳光,等.烟草黑胫病抗性的漂浮苗鉴定方法[J].中国烟草科学,2014,35(1):76-79.LI M Y,LIU Y,XIAO B G,et al.A float system resistant identification method of tobacco black shank[J].Chinese Tobacco Science,2014,35(1):76-79.
[10]周向平,杨全柳,王锡春,等.烟草品种(系)的烟草黑胫病抗性鉴定[J].作物研究,2013(6):637-639.ZHOU X P,YANG Q L,WANG X C,et al.Resistance evaluation of tobacco varieties and tobacco lines to tobacco black shank disease[J].Crop Research,2013(6):637-639.
[11]申莉莉.烟草突变体筛选与鉴定方法篇:2.烟草抗主要病虫害突变体的筛选与鉴定[J].中国烟草科学,2012,33(2):102-104.SHEN L L.Screening and identification of tobacco mutants:2.Screening and identification of tobacco resistance to major pests and diseases mutants[J].Chinese Tobacco Science,2012,33(2):102-104.
[12]李梅云,李永平,刘勇,等.抗黑胫病烟草种质资源的田间筛选[J].云南农业大学学报(自然科学版),2011,26(5):725-729.LI M Y,LI Y P,LIU Y,et al.Screening of resistant tobacco germplasm to black shank disease in field[J].Journal of Yunnan Agricultural University(Nat.Sci.Ed.),2011,26(5):725-729.
[13]王桥美,范静华,果志华,等.烟草黑胫病菌毒素对烟草防御性相关酶的诱导作用[J].云南农业大学学报,2011,26(1):20-25.WANG Q M,FAN J H,GUO Z H,et al.Induction effect of toxin extracts of phytophthora nicotianae on tobacco defense-related enzyme activities[J].Journal of Yunnan Agricultural University,2011,26(1):20-25.
[14]时焦,孙丽萍,孟坤,等.一种培养皿中筛选烟草抗赤星病种质材料的方法:中国,102876767 B[P].2013.01.16.SHI J,SUN L P,MENG K,et al.A method for screening tobacco anti-amazing disease germplasm material in culture dish:CN,102876767 B[P].2013.01.16.
[15]丁燕芳,平文丽,孙计平,等.烟草黑胫病抗性鉴定方法比较及其相关性分析[J].湖北农业科学,2017,56(18):3477-3480.DING Y F,PING W L,SUN J P,et al.Comparison of methods to identify resistance to tobacco black shank disease and their correlation analysis[J].Hubei Agricultural Sciences,2017,56(18):3477-3480.
[16]ZHANG Y S,GUO X,YAN X X,et al.Identification of stably expressed QTL for resistance to black shank disease in tobacco(Nicotiana tabacum L.)line Beinhart1000-1[J].The Crop Journal,2018,6(3):282-290.
[17]庞龙,黄河,周嘉平.烟草黑胫病菌毒素的分离纯化与性质的研究[J].科学通报,1986,31(18):1416-1416.PANG L,HUANG H,ZHOU J P.Isolation,purification and properties of toxin from tobacco black shank diseases[J].Science Bulletin,1986,31(18):1416-1420.
[18]高俊凤.植物生理学实验指导[M].北京:高等教育出版社,2006:140-148.GAO J F.Plant Physiology Experiment Guide[M].Beijing:Higher Education Press,2006:140-148.
[19]胡重怡,郑少清,陈兴江,等.烟草无菌苗培养前的种子消毒技术研究[J].中国烟草科学,2007,28(2):45-51.HU Z Y,ZHENG S Q,CHEN X J,et al.Methods for seed sterilization before aseptic tobacco seedling culture[J].Chinese Tobacco Science,2007,28(2):45-51.
[20]晏娟,黎定军.筛选烟草抗黑胫病细胞突变体的粗毒素浓度的初步研究[J].湖南农业科学,2011(5):66-68.YAN J,LI D J.A primary study on screening crude toxin concentration for cell mutants of tobacco against black shank[J].Hunan Agricultural Sciences,2011(5):66-68.
[21]高崇,吴国贺,李佰霖,等.延边烟区烟草菌核病病原菌鉴定及抗性种质筛选[J].中国烟草科学,2018,39(2):69-75.GAO C,WU G H,LI B L,et al.Identification of tobacco sclerotinia rot in Yanbian and screening for resistant germplasms[J].Chinese Tobacco Science,2018,39(2):69-75.
[22]叶协锋,张晓帆,郑宪滨,等.复合盐碱处理下烤烟品种发芽特性及耐盐性评价[J].中国烟草科学,2017,38(3):37-43.YE X F,ZHANG X F,ZHENG X B,et al.Germination characteristics of flue-cured tobacco varieties under mixed salt-alkali stresses and evaluations of their saline-alkali tolerance[J].Chinese Tobacco Science,2017,38(3):37-43.