Effects of Seed Viability and Number of Generations on Genetic Integrity of Soybean Germplasm Zhonghuang 18 by AFLP Markers
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摘要
The seeds of a soybean cultivar Zhonghuang 18 were subjected to accelerated aging for 0(population G_0-1), 112(population G_0-2), 154(population G_0-3) and 196 d(population G_0-4), whose germination percentage was found to be 98.0%, 95.0%, 81.0%, and 79.0%, respectively. Then, the four populations were regenerated twice in the field. The first descendant populations were marked as G_1-1, G_1-2, G_1-3 and G_1-4, and the second were marked as G_2-1, G_2-2, G_2-3 and G_2-4, respectively. The genetic variation between the control population(G_0-1) and the experimental populations was analyzed using 12 AFLP primer combinations. The results showed that there was no significant difference in genetic similarity between the 11 experimental populations and the control population G_0-1. The genetic similarity between population G_2-4 and G_0-1 was still as high as 0.933 3, indicating that the F_2 generation of the population whose germination percentage was only 79.0% still had a high genetic similarity to the control population. The results of t-tests revealed that the populations G_1-1, G_2-1, G_1-2 and G_2-2 showed no significant difference from the control population G_0-1 in effective number of alleles per locus(Ae), genetic diversity index(H) and Shannon's diversity index(I), while these indices of populations G_0-3, G_0-4, G_1-3, G_1-4, G_2-3 and G_2-4 were significantly reduced. χ~2 tests indicated that the populations G_1-1 and G_2-1 showed little difference, and the populations G_0-2, G_0-3, G_0-4, G_1-2, G_1-3, G_1-4, G_2-2, G_2-3, and G_2-4 showed great difference in allele frequency distribution from the control population G_0-1, and the difference was greater when the seed viability was lower. Compared with the control population G_0-1, the number of rare alleles(Nr) of the populations G_0-2, G_1-1, G_2-1, G_1-2 and G_2-2 showed no significant difference, while that of the populations G_0-3, G_0-4, G_1-3, G_1-4, G_2-3 and G_2-4 declined obviously. These results revealed that compared with the control population, the genetic diversity and Nr for the descendant populations of the populations with 98.0% and 95.0% germination percentages did not change significantly, but declined greatly for the descendant populations of the populations with 81.0.% and 79.0% percentages. The results suggested that the decline in seed viability has a greater impact than the number of generations on genetic structure of soybean germplasm. So, it is suggested that soybean seed with an initial germination percentage of 98.0% should be regenerated before its germination percentage declines to 81.0%.
The seeds of a soybean cultivar Zhonghuang 18 were subjected to accelerated aging for 0(population G_0-1), 112(population G_0-2), 154(population G_0-3) and 196 d(population G_0-4), whose germination percentage was found to be 98.0%, 95.0%, 81.0%, and 79.0%, respectively. Then, the four populations were regenerated twice in the field. The first descendant populations were marked as G_1-1, G_1-2, G_1-3 and G_1-4, and the second were marked as G_2-1, G_2-2, G_2-3 and G_2-4, respectively. The genetic variation between the control population(G_0-1) and the experimental populations was analyzed using 12 AFLP primer combinations. The results showed that there was no significant difference in genetic similarity between the 11 experimental populations and the control population G_0-1. The genetic similarity between population G_2-4 and G_0-1 was still as high as 0.933 3, indicating that the F_2 generation of the population whose germination percentage was only 79.0% still had a high genetic similarity to the control population. The results of t-tests revealed that the populations G_1-1, G_2-1, G_1-2 and G_2-2 showed no significant difference from the control population G_0-1 in effective number of alleles per locus(Ae), genetic diversity index(H) and Shannon's diversity index(I), while these indices of populations G_0-3, G_0-4, G_1-3, G_1-4, G_2-3 and G_2-4 were significantly reduced. χ~2 tests indicated that the populations G_1-1 and G_2-1 showed little difference, and the populations G_0-2, G_0-3, G_0-4, G_1-2, G_1-3, G_1-4, G_2-2, G_2-3, and G_2-4 showed great difference in allele frequency distribution from the control population G_0-1, and the difference was greater when the seed viability was lower. Compared with the control population G_0-1, the number of rare alleles(Nr) of the populations G_0-2, G_1-1, G_2-1, G_1-2 and G_2-2 showed no significant difference, while that of the populations G_0-3, G_0-4, G_1-3, G_1-4, G_2-3 and G_2-4 declined obviously. These results revealed that compared with the control population, the genetic diversity and Nr for the descendant populations of the populations with 98.0% and 95.0% germination percentages did not change significantly, but declined greatly for the descendant populations of the populations with 81.0.% and 79.0% percentages. The results suggested that the decline in seed viability has a greater impact than the number of generations on genetic structure of soybean germplasm. So, it is suggested that soybean seed with an initial germination percentage of 98.0% should be regenerated before its germination percentage declines to 81.0%.
The seeds of a soybean cultivar Zhonghuang 18 were subjected to accelerated aging for 0(population G_0-1), 112(population G_0-2), 154(population G_0-3) and 196 d(population G_0-4), whose germination percentage was found to be 98.0%, 95.0%, 81.0%, and 79.0%, respectively. Then, the four populations were regenerated twice in the field. The first descendant populations were marked as G_1-1, G_1-2, G_1-3 and G_1-4, and the second were marked as G_2-1, G_2-2, G_2-3 and G_2-4, respectively. The genetic variation between the control population(G_0-1) and the experimental populations was analyzed using 12 AFLP primer combinations. The results showed that there was no significant difference in genetic similarity between the 11 experimental populations and the control population G_0-1. The genetic similarity between population G_2-4 and G_0-1 was still as high as 0.933 3, indicating that the F_2 generation of the population whose germination percentage was only 79.0% still had a high genetic similarity to the control population. The results of t-tests revealed that the populations G_1-1, G_2-1, G_1-2 and G_2-2 showed no significant difference from the control population G_0-1 in effective number of alleles per locus(Ae), genetic diversity index(H) and Shannon's diversity index(I), while these indices of populations G_0-3, G_0-4, G_1-3, G_1-4, G_2-3 and G_2-4 were significantly reduced. χ~2 tests indicated that the populations G_1-1 and G_2-1 showed little difference, and the populations G_0-2, G_0-3, G_0-4, G_1-2, G_1-3, G_1-4, G_2-2, G_2-3, and G_2-4 showed great difference in allele frequency distribution from the control population G_0-1, and the difference was greater when the seed viability was lower. Compared with the control population G_0-1, the number of rare alleles(Nr) of the populations G_0-2, G_1-1, G_2-1, G_1-2 and G_2-2 showed no significant difference, while that of the populations G_0-3, G_0-4, G_1-3, G_1-4, G_2-3 and G_2-4 declined obviously. These results revealed that compared with the control population, the genetic diversity and Nr for the descendant populations of the populations with 98.0% and 95.0% germination percentages did not change significantly, but declined greatly for the descendant populations of the populations with 81.0.% and 79.0% percentages. The results suggested that the decline in seed viability has a greater impact than the number of generations on genetic structure of soybean germplasm. So, it is suggested that soybean seed with an initial germination percentage of 98.0% should be regenerated before its germination percentage declines to 81.0%.
引文
[1]The Food and Agriculture Organization of the United Nations(FAO).Report on the state of the world's plants genetic resources for food and agriculture[R].Rome,Italy:FAO.1996.
[2]WANG SM,LI LH,LI Y,et al.Status of plant genetic resources for food and agricultural in China(I)[J].Journal of Plant Genetic Resources,2011,12(1):1-12.
[3]LU XX,CUI CS,CHEN XL,et al.Survey of seed germinability after10-12 years storage in the national genebank of China[J].Plant Genet Resour Sci,2001,2(2):1-5.
[4]WALTERS C,WHEELER LM,GROTENHUIS JM.Longevity of seeds stored in a Genebank:Species characteristics[J].Seed Sci Res,2005,15:1-20.
[5]QIU LJ,CHANG RZ,CHEN KM,et al.Analysis of conservation and regeneration statues for Chinese soybean germplasm[J].Plant Genet Resour Sci,2002,3(2):34-39.
[6]Frankel OH,Brown AHD,Burdon JJ.The conservation of plant biodiversity[M].Cambridge:Cambridge University Press,1995.
[7]VAN HINTUM TJL,VAN DE WIEL CCM,VISSER DL,et al.The distribution of genetic diversity in a Brassica oleracea gene bank collection related to the effects on diversity of regeneration,as measured with AFLPs[J].Theor Appl Genet,1984,114:777-786.
[8]ROOS EE.Genetic shifts in mixed bean populations.I.Storage effects[J].Crop Sci,1984,24:240-244.
[9]ROOS EE.Report of the storage committee working population on effects of storage on genetic integrity 1980-1983[J].Seed Sci Technol,1984,12:255-260.
[10]BREESE EL.Regeneration and multiplication of germplasm resources in seed genebanks:The scientific background.Rome:IBPGR,1989.
[11]STOYANOVA SD.Effect of seed aging and regeneration on the genetic composition of wheat[J].Seed Sci Technol,1992,20:489-496.
[12]VAN HINTUM TH JL,VISSER DL.Duplication within and between germplasm collections:II.Duplication in four European barley collections[J].Genet Res Crop Evol,1995,42:135-145.
[13]YONEZAWA K,ISHII T,NOMURA T,et al.Effectiveness of some management procedures for seed regeneration of plant genetic resources accessions[J].Genetica,1996,43:517-524.
[14]RIO AH,BAMBERG JB,HUAMAN Z.Assessing changes in the genetic diversity of potato gene banks:I.Effects of seed increase[J].Theor Appl Genet,1997,95:191-198.
[15]RAO NK,BRAMEL PJ,REDDY KN,et al.Optimizing seed quality during germplasm regeneration in pearl millet[J].Genet Res Crop Evol,2002,49:153-157.
[16]CHEBOTAR S,RODER MS,KORZUN V,et al.Molecular studies on genetic integrity of open pollinating species rye(Secale cereale L.)after long term Genebank maintenance[J].Theor Appl Genet,2003,107(8):1469-1476.
[17]MURATA M,TSUCHIYA T,ROOS EE.Chromosome damage induced by artificial seed in barley[J].Theor Appl Genet,1984,67:161-170.
[18]DOURADO AM,ROBERTS EH.Chromosome aberrations induced during storage in barley and pea seeds[J].Ann Bot,1984,54:767-779.
[19]ROBERTS EH.Loss of viability:Chromosomal and genetic aspects[J].Seed Sci Technol,1973,1:515-527.
[20]ZHANG H,LU XX,ZHAGN ZE,et al.Effect of seed aging on change of genetic integrity in maize germplasm[J].Journal of Plant Genetic Resources,2005,6(3):271-275.
[21]TAO KL,PERINO P,PIERLUIGI L,et al.Rapid and nondestructive method for detecting composition change in wheat germplasm accessions[J].Crop Sci,1992,32:1039-1042.
[22]SOENGAS P,CARTEA E,LEMA M,et al.Effect of regeneration Procedures on the genetic integrity of Brassica oleracea accessions[J].Mol Breed,2009,23:389-395.
[23]XIA B,LU XX,CHE XL,et al.Detection of genetic integrity of soybean germplasm using SSR markers[J].Soybean Sci,2007,26(3):305-309.
[24]ZHANG YM,GAI JY.A study on suitable sample size in the conservation of landrace of soybeans[J].Sci Agric Sin,1995,28(S):70-75.
[25]ZHENG XY,GUO YP,MA EB.Development of AFLP molecular markers[J].Chem Life,2003,23(1):65-67.
[26]MAUGHAN PJ,SAGHAI-MAROOF MA,BUSS GR,et al.Amplified fragment length polymorphism(AFLP)in soybean species diversity,inheritance,and near-isogenic line analysis[J].Theor Appl Genet,2003,93(3):392-401.
[27]HU XR,LU XX,ZHANG YL,et al.Studies on the genetic integrity ultra dry seed of wheat with AFLP markers[J].J Plant Genet Resour,2003,4(2):162-165.
[28]FANG JH,LIU X,LU XX.Technical Regulation on Characterization and Documentation for Crop Germplasm Resources[M].Beijing:China Agriculture Press,2008.
[29]ISTA.International Rules For Seed Testing 1996[M].Beijing:China Agriculture Press,1996.
[30]TIAN QZ,GAI JY,YU DY,et al.A study on amplified fragment length polymorphism(AFLP)in soybean[J].Soybean Sci,2000,19(3):210-217.
[31]TIAN QZ,GAI JY,YU DY,et al.AFLP Fingerprint Analysis of G.soja and G.max in China[J].Sci Agric Sin,2001,34(5):465-468.
[32]SANGUINETTI CJ,DIAS NE,SIMPSON AJ.Rapid silver staining and recovery of PCR products separated on polyacrylamide gels[J].Biotechniques,1994,17:914-921.
[33]YEH FC,YANG RC,BOYLE T.POPGENE Version 1.31 quick user guide[Z].Canada:University of Alberta,and Center for International Forestry Research,1999.
[34]ROHLF F J.NTSYS-pc:numerical taxonomy and multivariate analysis system,Version 2.1,user guide[Z].New York:Exeter Publications,2000.
[35]St OYANOVA SD.Genetic shifts and variations of gliadins induced by seed aging[J].Seed Sci Technol,1991,19:363-371.
[36]PARZIES HK,SPOOR W,ENNOS RA.Genetic diversity of barley landrace accessions(Hordeum vulgare ssp.vulgare)conserved for different lengths of time in ex situ gene banks[J].Heredity,2000,84:476-486.
[37]ROOS EE.Genetic shifts in mixed bean populations.II.Effects of regeneration[J].Crop Sci,1984,24:711-715.
[38]BORNER A,CHEBOTAR S,KORZUN V.Molecular characterization of the genetic integrity of wheat(Triticum aestivum L.)germplasm after long-term maintenance[J].Theor Appl Genet,2000,100:494-497.
[39]MARSHALL DR,BROWN AHD.Optimum sampling strategies in genetic conservation[A].In:Frankel O H,Hawkes J G eds.Crop Genetic Resources for Today and Tomorrow[C].Cambridge:Cambridge University Press,1975.
[40]ROOS EE.Genetic changes in a collection over time[J].Hort Science,1988,23:86-90.
[41]CHWEDORZEWSKA KJ,BEDNAREK PT,PUCHALSKI J,et al.AFLP-profiling of long-term stored and regenerated rye genebank samples[J].Cell Mol Biol Lett,2002,7:457-463.
[42]CHWEDORZEWSKA KJ,BEDNAREK PT,PUCHALSKI J.Studies on changes in specific rye genome regions due to seed aging and regeneration[J].Cell Mol Biol Lett,2002,7:569-576.
[43]ZUREK G.Effect of seed storage on germplasm integrity of meadow fescue(Festuca pratensis Huds.)[J].Genet Res Crop Evol,1999,46:485-490.
[44]FAO/IPGRI.Genebank standards[Z].Rome:FAO/IPGRI,1994.
[45]LI LZ,WANG LN,GENG XL,et al.Seed vigor influences on some agronomic characteristics of soybean[J].J Hebei Agri Sci,2002,6(1):14-19.
[46]SACKVILLE HAMILTON N R,CHORLTON K H.Regeneration of accessions in seed collection[Z].Rome:IPGRI,1997.
[47]SCHOEN DJ,DAVID JL,BATAILLON TM.Deleterious mutation accumulation and the regeneration of genetic resources[J].Proc Natl Acad Sci USA,1998,95:394-399.
[48]LE CLERC V,BRIARD M,GRANGER J,et al.Genebank biodiversity assessments regarding optimal sample size and seed harvesting techniques for the regeneration of carrot accessions[J].Biodivers Conser,2003,12:2227-2336.
[2]WANG SM,LI LH,LI Y,et al.Status of plant genetic resources for food and agricultural in China(I)[J].Journal of Plant Genetic Resources,2011,12(1):1-12.
[3]LU XX,CUI CS,CHEN XL,et al.Survey of seed germinability after10-12 years storage in the national genebank of China[J].Plant Genet Resour Sci,2001,2(2):1-5.
[4]WALTERS C,WHEELER LM,GROTENHUIS JM.Longevity of seeds stored in a Genebank:Species characteristics[J].Seed Sci Res,2005,15:1-20.
[5]QIU LJ,CHANG RZ,CHEN KM,et al.Analysis of conservation and regeneration statues for Chinese soybean germplasm[J].Plant Genet Resour Sci,2002,3(2):34-39.
[6]Frankel OH,Brown AHD,Burdon JJ.The conservation of plant biodiversity[M].Cambridge:Cambridge University Press,1995.
[7]VAN HINTUM TJL,VAN DE WIEL CCM,VISSER DL,et al.The distribution of genetic diversity in a Brassica oleracea gene bank collection related to the effects on diversity of regeneration,as measured with AFLPs[J].Theor Appl Genet,1984,114:777-786.
[8]ROOS EE.Genetic shifts in mixed bean populations.I.Storage effects[J].Crop Sci,1984,24:240-244.
[9]ROOS EE.Report of the storage committee working population on effects of storage on genetic integrity 1980-1983[J].Seed Sci Technol,1984,12:255-260.
[10]BREESE EL.Regeneration and multiplication of germplasm resources in seed genebanks:The scientific background.Rome:IBPGR,1989.
[11]STOYANOVA SD.Effect of seed aging and regeneration on the genetic composition of wheat[J].Seed Sci Technol,1992,20:489-496.
[12]VAN HINTUM TH JL,VISSER DL.Duplication within and between germplasm collections:II.Duplication in four European barley collections[J].Genet Res Crop Evol,1995,42:135-145.
[13]YONEZAWA K,ISHII T,NOMURA T,et al.Effectiveness of some management procedures for seed regeneration of plant genetic resources accessions[J].Genetica,1996,43:517-524.
[14]RIO AH,BAMBERG JB,HUAMAN Z.Assessing changes in the genetic diversity of potato gene banks:I.Effects of seed increase[J].Theor Appl Genet,1997,95:191-198.
[15]RAO NK,BRAMEL PJ,REDDY KN,et al.Optimizing seed quality during germplasm regeneration in pearl millet[J].Genet Res Crop Evol,2002,49:153-157.
[16]CHEBOTAR S,RODER MS,KORZUN V,et al.Molecular studies on genetic integrity of open pollinating species rye(Secale cereale L.)after long term Genebank maintenance[J].Theor Appl Genet,2003,107(8):1469-1476.
[17]MURATA M,TSUCHIYA T,ROOS EE.Chromosome damage induced by artificial seed in barley[J].Theor Appl Genet,1984,67:161-170.
[18]DOURADO AM,ROBERTS EH.Chromosome aberrations induced during storage in barley and pea seeds[J].Ann Bot,1984,54:767-779.
[19]ROBERTS EH.Loss of viability:Chromosomal and genetic aspects[J].Seed Sci Technol,1973,1:515-527.
[20]ZHANG H,LU XX,ZHAGN ZE,et al.Effect of seed aging on change of genetic integrity in maize germplasm[J].Journal of Plant Genetic Resources,2005,6(3):271-275.
[21]TAO KL,PERINO P,PIERLUIGI L,et al.Rapid and nondestructive method for detecting composition change in wheat germplasm accessions[J].Crop Sci,1992,32:1039-1042.
[22]SOENGAS P,CARTEA E,LEMA M,et al.Effect of regeneration Procedures on the genetic integrity of Brassica oleracea accessions[J].Mol Breed,2009,23:389-395.
[23]XIA B,LU XX,CHE XL,et al.Detection of genetic integrity of soybean germplasm using SSR markers[J].Soybean Sci,2007,26(3):305-309.
[24]ZHANG YM,GAI JY.A study on suitable sample size in the conservation of landrace of soybeans[J].Sci Agric Sin,1995,28(S):70-75.
[25]ZHENG XY,GUO YP,MA EB.Development of AFLP molecular markers[J].Chem Life,2003,23(1):65-67.
[26]MAUGHAN PJ,SAGHAI-MAROOF MA,BUSS GR,et al.Amplified fragment length polymorphism(AFLP)in soybean species diversity,inheritance,and near-isogenic line analysis[J].Theor Appl Genet,2003,93(3):392-401.
[27]HU XR,LU XX,ZHANG YL,et al.Studies on the genetic integrity ultra dry seed of wheat with AFLP markers[J].J Plant Genet Resour,2003,4(2):162-165.
[28]FANG JH,LIU X,LU XX.Technical Regulation on Characterization and Documentation for Crop Germplasm Resources[M].Beijing:China Agriculture Press,2008.
[29]ISTA.International Rules For Seed Testing 1996[M].Beijing:China Agriculture Press,1996.
[30]TIAN QZ,GAI JY,YU DY,et al.A study on amplified fragment length polymorphism(AFLP)in soybean[J].Soybean Sci,2000,19(3):210-217.
[31]TIAN QZ,GAI JY,YU DY,et al.AFLP Fingerprint Analysis of G.soja and G.max in China[J].Sci Agric Sin,2001,34(5):465-468.
[32]SANGUINETTI CJ,DIAS NE,SIMPSON AJ.Rapid silver staining and recovery of PCR products separated on polyacrylamide gels[J].Biotechniques,1994,17:914-921.
[33]YEH FC,YANG RC,BOYLE T.POPGENE Version 1.31 quick user guide[Z].Canada:University of Alberta,and Center for International Forestry Research,1999.
[34]ROHLF F J.NTSYS-pc:numerical taxonomy and multivariate analysis system,Version 2.1,user guide[Z].New York:Exeter Publications,2000.
[35]St OYANOVA SD.Genetic shifts and variations of gliadins induced by seed aging[J].Seed Sci Technol,1991,19:363-371.
[36]PARZIES HK,SPOOR W,ENNOS RA.Genetic diversity of barley landrace accessions(Hordeum vulgare ssp.vulgare)conserved for different lengths of time in ex situ gene banks[J].Heredity,2000,84:476-486.
[37]ROOS EE.Genetic shifts in mixed bean populations.II.Effects of regeneration[J].Crop Sci,1984,24:711-715.
[38]BORNER A,CHEBOTAR S,KORZUN V.Molecular characterization of the genetic integrity of wheat(Triticum aestivum L.)germplasm after long-term maintenance[J].Theor Appl Genet,2000,100:494-497.
[39]MARSHALL DR,BROWN AHD.Optimum sampling strategies in genetic conservation[A].In:Frankel O H,Hawkes J G eds.Crop Genetic Resources for Today and Tomorrow[C].Cambridge:Cambridge University Press,1975.
[40]ROOS EE.Genetic changes in a collection over time[J].Hort Science,1988,23:86-90.
[41]CHWEDORZEWSKA KJ,BEDNAREK PT,PUCHALSKI J,et al.AFLP-profiling of long-term stored and regenerated rye genebank samples[J].Cell Mol Biol Lett,2002,7:457-463.
[42]CHWEDORZEWSKA KJ,BEDNAREK PT,PUCHALSKI J.Studies on changes in specific rye genome regions due to seed aging and regeneration[J].Cell Mol Biol Lett,2002,7:569-576.
[43]ZUREK G.Effect of seed storage on germplasm integrity of meadow fescue(Festuca pratensis Huds.)[J].Genet Res Crop Evol,1999,46:485-490.
[44]FAO/IPGRI.Genebank standards[Z].Rome:FAO/IPGRI,1994.
[45]LI LZ,WANG LN,GENG XL,et al.Seed vigor influences on some agronomic characteristics of soybean[J].J Hebei Agri Sci,2002,6(1):14-19.
[46]SACKVILLE HAMILTON N R,CHORLTON K H.Regeneration of accessions in seed collection[Z].Rome:IPGRI,1997.
[47]SCHOEN DJ,DAVID JL,BATAILLON TM.Deleterious mutation accumulation and the regeneration of genetic resources[J].Proc Natl Acad Sci USA,1998,95:394-399.
[48]LE CLERC V,BRIARD M,GRANGER J,et al.Genebank biodiversity assessments regarding optimal sample size and seed harvesting techniques for the regeneration of carrot accessions[J].Biodivers Conser,2003,12:2227-2336.