老化处理对老芒麦种子活力及醇溶蛋白组成的影响
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摘要
为揭示人工老化处理对老芒麦种子活力及醇溶蛋白组成与表达量的影响,以老芒麦(Elymussibiricus)种子为试验材料,用高温(45℃)、高湿(100%RH)的方法处理种子,对不同老化程度的种子进行发芽实验并计算相应指数,并对酸性聚丙烯酰氨凝胶电泳(A-PAGE)检测醇溶蛋白的体系进行优化。结果表明:人工老化处理7天后,种子的发芽率、发芽势、发芽指数和活力指数依次下降58%、56.34%、75.36%、83%。从凝胶电泳体系优化结果可以看出,最优体系为:300 V电压、上样量为10μL、18%的聚丙烯酰氨凝胶。老化后的种子ω区醇溶蛋白出现了带型的变化,即原有谱带的丢失、新谱带的出现。由此可以看出种子活力指数对老化处理最为敏感,它的下降与电导率的变化相关性较大,种子活力丧失与原有谱带的丢失、新谱带的出现在同一时间,说明它与ω区域醇溶蛋白的变化有关。
To reveal the effects of artificial aging on seed vigor, and constitutes and expression of prolamin, the Elymus sibiricus seeds were used as the tested materials. The seeds were treated with the method of high temperature(45℃) and high humidity(100% RH). The germination experiment was conducted on the seeds with different artificial aging degrees and the corresponding germination indexes were calculated. The acidacrylamide gel electrophoresis(A-PAGE) detecting prolamin was optimized at the same time. The results showed that the seed germination rate, germination potential, germination index, and vigor index decreased by58%, 56.34%, 75.36%, and 83% after artificial aging for seven days. According to the results of gel electrophoresis system, the optimal system was determined as 300 V voltage, 10 μL loading volume, and 18%polyacrylamide gel. The band of prolamin in the ω-zones changed after artificial aging, such as the absence of the original bands and the appearance of new bands. Therefore, the seed vigor index is the most sensitive indicator to aging treatment, and the decrease of seed vigor is related to the change of conductivity. And the lose of seed vigor is companied with the absence of the original band and the appearance of new bands,indicating that the seed vigor is related to the changes of ω-prolamin.
To reveal the effects of artificial aging on seed vigor, and constitutes and expression of prolamin, the Elymus sibiricus seeds were used as the tested materials. The seeds were treated with the method of high temperature(45℃) and high humidity(100% RH). The germination experiment was conducted on the seeds with different artificial aging degrees and the corresponding germination indexes were calculated. The acidacrylamide gel electrophoresis(A-PAGE) detecting prolamin was optimized at the same time. The results showed that the seed germination rate, germination potential, germination index, and vigor index decreased by58%, 56.34%, 75.36%, and 83% after artificial aging for seven days. According to the results of gel electrophoresis system, the optimal system was determined as 300 V voltage, 10 μL loading volume, and 18%polyacrylamide gel. The band of prolamin in the ω-zones changed after artificial aging, such as the absence of the original bands and the appearance of new bands. Therefore, the seed vigor index is the most sensitive indicator to aging treatment, and the decrease of seed vigor is related to the change of conductivity. And the lose of seed vigor is companied with the absence of the original band and the appearance of new bands,indicating that the seed vigor is related to the changes of ω-prolamin.
引文
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[15]国际种子检验协会(ISTA).种苗评定与种子活力测定方法手册[M].北京:北京农业大学出版社,1987.
[16]耿以礼.中国主要植物图说——禾本科[M].北京:科学出版社,1959:421-446.
[17]张自阳,姜小苓,王娟娟,等.人工老化处理对小麦种子活力和醇溶蛋白、麦谷蛋白组成的影响[J].河南农业科学,2014,43(2):24-27.
[18]董新红,宋明.种子劣变的原因及其防止与修复[J].中国种业,2002(1):39-40.
[19] Basavarajappa B S, Shekar Shetty H, Prakash H S. Membranedeterioration and other biochemical-changes, associated withaccelerated aging of maize seeds[J]. Seed Science and Technology,1991,19(2):279-286.
[20]王继红,陈绍宁,白冰,等.人工老化对玉米种子蛋白质组的影响[J].河南农业大学学报,2009,43(3):232-235.
[21]张晗,卢新雄,张志娥,等.种子老化诱导小麦染色体畸变及大麦醇溶蛋白带型频率变化的研究[J].植物遗传资源学报,2004(1):56-61.
[22] Kalpana R, Madhava R A O. On the ageing mechanism inpigeonpea(Cajanus cajan(L.)Millsp)seeds[J]. Seed Science andTechnology, 1995,23(1):1-9.
[23]董贵俊.向日葵(Helianthus annuus L.)种质资源相关的基础研究[D].北京:中国科学院研究生院(植物研究所),2006.
[24] Rajjou L. Gallardo K, Debeaujon L, et al. The effect ofα-amanitinon the Arabidopsis seed proteomc highlights the distinct roles ofstored and ncosynthesized mRNAs during germination[J].PlantPhysiology,2004,134(4):1593-1613.
[25] Parzies H K. Spoor W, Ennos R A. Genetic diversity of barleylandraee accessions conserved for different lengths of time in cxsitu gene bank.Heredity,2000,84(4):476-486.
[26]杨剑平,刘春辉,王海涛,等.人工老化处理对小麦种胚及幼苗生长的影响[J].北京农学院学报,1995(1):17-21.
[27]梁海荣,邢侗姬,周小梅.不同活力的小麦种子生理生化及其修复的研究[J].山西大学学报:自然科学版,2010(2):286-290.
[28]吴浩,周青平,颜红波,等.垂穗披碱草种子老化过程中生理生化特性的研究[J].青海大学学报:自然科学版,2014,32(3):6-10.
[29] Liu A R, Zhao K F. The accumulation and osmotic regulation of saltmustard osmotic regulation under salt stress. Journal of PlantPhysiology and Molecular Biology, 2005,31(4):389-39.
[2]刘军,黄上志,傅家瑞,等.种子活力与蛋白质关系的研究进展[J].植物学通报,2001(1):45,46-51.
[3]王继红,陈绍宁,白冰,等.人工老化对玉米种子蛋白质组的影响[J].河南农业大学学报,2009,43(3):232-235.
[4]周峰.种子贮藏蛋白质丰度研究进展[J].河南农业科学,2015,44(1):1-3,76.
[5]舒英杰,陶源,王爽,等.高等植物种子活力的生物学研究进展[J].西北植物学报,2013,33(8):1709-1716.
[6]张自阳,马景周,王智煜,等.人工老化对小麦种子活力及醇溶蛋白组成的影响[J].种子,2017,36(8):42-47.
[7] Tiedemann J, Neubohn B, Muntz K. Different functions of vicilinand legumin are reflected in the histopaettern of globulinmobilization during germination of vetch[J]. Planta,2000,211(1):1-12.
[8] Tiedemann J, Schlereth A, Muntz K. Different tissue-specificexpression of cysteine proteinases forms the basis for the fine-tunedmobilization of storage globulin during and after germination inlegume seeds[J].Planta,2001,212(5):7-16.
[9]董鸿运,班青,乔玉玲.杉木、马尾松种子人工老化过程中某些生理生化变化规律的研究[J].种子,1987(2):14-18.
[10] Draper S R. ISTA Variety committee Report of the Working Groupfor Biochemical Tests for Cultivar Identification1987-1986[J].SeedSci Technol,1987(15):431-434.
[11] Kawaura K, Mochida K, Ogihara Y. Expression profile of twostorage-protein gene families in hexaploidy wheat revealed by large-scale analysis of expressed sequence tags[J]. Plant Physiol,2005,139:1870-1880.
[12]颜启传.种子学[M].北京:中国农业出版社,2001:110-431.
[13] Abdul-BakiA A. Biochemic alaspects of seed vigor[J].Hortscience,1980,15(3):735-771.
[14]国家质量技术监督局.中华人民共和国国家标准牧草种子检验规程(GB/T 2930.1-11-2001)[M].北京:中国标准出版社, 2001,6:31-48.
[15]国际种子检验协会(ISTA).种苗评定与种子活力测定方法手册[M].北京:北京农业大学出版社,1987.
[16]耿以礼.中国主要植物图说——禾本科[M].北京:科学出版社,1959:421-446.
[17]张自阳,姜小苓,王娟娟,等.人工老化处理对小麦种子活力和醇溶蛋白、麦谷蛋白组成的影响[J].河南农业科学,2014,43(2):24-27.
[18]董新红,宋明.种子劣变的原因及其防止与修复[J].中国种业,2002(1):39-40.
[19] Basavarajappa B S, Shekar Shetty H, Prakash H S. Membranedeterioration and other biochemical-changes, associated withaccelerated aging of maize seeds[J]. Seed Science and Technology,1991,19(2):279-286.
[20]王继红,陈绍宁,白冰,等.人工老化对玉米种子蛋白质组的影响[J].河南农业大学学报,2009,43(3):232-235.
[21]张晗,卢新雄,张志娥,等.种子老化诱导小麦染色体畸变及大麦醇溶蛋白带型频率变化的研究[J].植物遗传资源学报,2004(1):56-61.
[22] Kalpana R, Madhava R A O. On the ageing mechanism inpigeonpea(Cajanus cajan(L.)Millsp)seeds[J]. Seed Science andTechnology, 1995,23(1):1-9.
[23]董贵俊.向日葵(Helianthus annuus L.)种质资源相关的基础研究[D].北京:中国科学院研究生院(植物研究所),2006.
[24] Rajjou L. Gallardo K, Debeaujon L, et al. The effect ofα-amanitinon the Arabidopsis seed proteomc highlights the distinct roles ofstored and ncosynthesized mRNAs during germination[J].PlantPhysiology,2004,134(4):1593-1613.
[25] Parzies H K. Spoor W, Ennos R A. Genetic diversity of barleylandraee accessions conserved for different lengths of time in cxsitu gene bank.Heredity,2000,84(4):476-486.
[26]杨剑平,刘春辉,王海涛,等.人工老化处理对小麦种胚及幼苗生长的影响[J].北京农学院学报,1995(1):17-21.
[27]梁海荣,邢侗姬,周小梅.不同活力的小麦种子生理生化及其修复的研究[J].山西大学学报:自然科学版,2010(2):286-290.
[28]吴浩,周青平,颜红波,等.垂穗披碱草种子老化过程中生理生化特性的研究[J].青海大学学报:自然科学版,2014,32(3):6-10.
[29] Liu A R, Zhao K F. The accumulation and osmotic regulation of saltmustard osmotic regulation under salt stress. Journal of PlantPhysiology and Molecular Biology, 2005,31(4):389-39.