老芒麦种子离区酶活变化及组织学分析
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摘要
通过对青藏高原东北缘6种野生老芒麦不同时期种子离区组织结构的观察和细胞壁水解酶活性的测定,在组织学和生理学层面阐释老芒麦种子落粒的成因。结果表明,老芒麦种子离区结构的形成时期早于种子脱落,离层由2~3层小体积、椭圆形的细胞紧凑排列组成且这些细胞木质化程度高于周围组织细胞,低落粒材料离层木质化细胞数量多于高落粒材料;低落粒材料在种子成熟期无明显的离层断裂,其断裂面粗糙,离层结构完整,而高落粒材料在种子成熟期出现明显的离层断裂,断裂面光滑且离层结构不完整;6种材料纤维素酶(CE)与多聚半乳糖醛酸酶(PG)活性的变化趋势在种子发育各时期均有差异,落粒关键期(抽穗后4~5周)这两种酶活性均与种子脱落密切相关;供试材料在落粒关键期CE与PG活性排序一致,且与同时期电镜观察结果相近;高落粒材料XH09在落粒关键期的两种酶活性均为最高(XH09CE=479.52IU·L-1,XH09PG=188.87pg·mL-1),而低落粒材料ZHN03两种酶活性均为最低。研究表明,老芒麦高落粒与低落粒材料在离层木质化程度、断裂面光滑度、离层结构完整性及落粒关键期细胞壁水解酶活性等方面具有差异。同时,离层木质化程度与落粒关键期细胞壁水解酶活性是造成野生老芒麦种子脱落的重要因素。
Histological and enzyme activity analysis of the abscission zone of seed from six Elymus sibiricus accessions from the northeast margin of the Qinghai-Tibet Plateau at different stages of seed maturation was undertaken.The results showed that the abscission layer of seeds were present before seed shattering.The abscission layer was composed of 2 to 3 layers of small volume cells,oval in shape and with a compact arrangement.The degree of lignification of these cells was higher than surrounding cells.The lignified cell number of low seed shattering accessions was higher than high seed shattering accessions;low seed shattering accessions did not have an abscission layer at the seed maturity stage.These accessions had a rough fracture surface and an integrated abscission layer structure.High seed shattering accessions developed a clear fracture or abscission layer during seed maturation.These materials had a smooth fracture surface and incomplete abscission layer structure.Changes in cellulase(CE)and polygalacturonase(PG)activity among accessions differed at different seed developmental stages.The activities of these cell wall hydrolases were both closely related to seed shattering;the CE and PG activity of high seed shattering accessions at the time of shattering was high and consistent with electron microscopy observations.The high seed shattering accession(XH09)possessed the highest enzyme activity(XH09 CE=479.52 IU·L-1,XH09 PG=188.87 pg·mL-1)during the seed shattering period while the low seed shattering accession(ZHN03)had the lowest enzyme activity.This study showed that there were clear differences in the degree of lignification of the abscission layer,fracture surface smoothness,structural integrity of the abscission layer and the activities of cell wall hydrolases between low and high seed shattering accessions during the critical period of seed shattering.The degree of lignification of the abscission layer and activities of cell wall hydrolases at the seed shattering stage were the most important factors influencing seed shattering in wild E.sibiricus accessions.
Histological and enzyme activity analysis of the abscission zone of seed from six Elymus sibiricus accessions from the northeast margin of the Qinghai-Tibet Plateau at different stages of seed maturation was undertaken.The results showed that the abscission layer of seeds were present before seed shattering.The abscission layer was composed of 2 to 3 layers of small volume cells,oval in shape and with a compact arrangement.The degree of lignification of these cells was higher than surrounding cells.The lignified cell number of low seed shattering accessions was higher than high seed shattering accessions;low seed shattering accessions did not have an abscission layer at the seed maturity stage.These accessions had a rough fracture surface and an integrated abscission layer structure.High seed shattering accessions developed a clear fracture or abscission layer during seed maturation.These materials had a smooth fracture surface and incomplete abscission layer structure.Changes in cellulase(CE)and polygalacturonase(PG)activity among accessions differed at different seed developmental stages.The activities of these cell wall hydrolases were both closely related to seed shattering;the CE and PG activity of high seed shattering accessions at the time of shattering was high and consistent with electron microscopy observations.The high seed shattering accession(XH09)possessed the highest enzyme activity(XH09 CE=479.52 IU·L-1,XH09 PG=188.87 pg·mL-1)during the seed shattering period while the low seed shattering accession(ZHN03)had the lowest enzyme activity.This study showed that there were clear differences in the degree of lignification of the abscission layer,fracture surface smoothness,structural integrity of the abscission layer and the activities of cell wall hydrolases between low and high seed shattering accessions during the critical period of seed shattering.The degree of lignification of the abscission layer and activities of cell wall hydrolases at the seed shattering stage were the most important factors influencing seed shattering in wild E.sibiricus accessions.
引文
[1]Dewey D R.Cytogenetics of Elymus sibiricus and its hybrids with Agropyron tauri,Elymus canadensis,and Agropyron caninum.International Journal of Plant Sciences,1974,135(1):80-87.
[2]Chen T,Nan Z B.Seed-borne fungi infection of Siberian wildrye:effects on seed germination and seedling growth.Acta Prataculturae Sinica,2015,24(2):96-103.陈焘,南志标.不同储存年限老芒麦种子种带真菌检测及致病性测定.草业学报,2015,24(2):96-103.
[3]Gu X Y,Guo Z H,Zhang X Q,et al.Genetic diversity of Elymus sibiricus germplasm resources revealed by SRAP markers.Acta Prataculturae Sinica,2014,23(1):205-216.顾晓燕,郭志慧,张新全,等.老芒麦种质资源遗传多样性的SRAP分析.草业学报,2014,23(1):205-216.
[4]Xie W,Zhao X,Zhang J,et al.Assessment of genetic diversity of Siberian wild rye(Elymus sibiricus L.)germplasms with variation of seed shattering and implication for future genetic improvement.Biochemical Systematics and Ecology,2015,58:211-218.
[5]Yan J J,Bai S Q,Ma X,et al.Genetic diversity of Elymus sibiricus and its breeding in China.Chinese Bulletin of Botany,2007,24(2):226-231.鄢家俊,白史且,马啸,等.老芒麦遗传多样性及育种研究进展.植物学报,2007,24(2):226-231.
[6]Konishi S,Izawa T,Lin S Y,et al.An SNP caused loss of seed shattering during rice domestication.Science,2006,312:1392-1396.
[7]Lin Z W,Li X R,Shannon L M,et al.Parallel domestication of the Shattering1genes in cereals.Nature Genetics,2012,44(6):720-724.
[8]Simons K J,Fellers J P,Trick H N,et al.Molecular characterization of the major wheat domestication gene Q.Genetics,2006,172(1):547-555.
[9]Kandemir N,Kudrna D A,Ullrich S E,et al.Molecular marker assisted genetic analysis of head shattering in six-rowed barley.Theoretical and Applied Genetics,2000,101(1):203-210.
[10]Pourkheirandish M,Hensel G,Kilian B,et al.Evolution of the grain dispersal system in barley.Cell,2015,162(3):527-539.
[11]Ji H S,Chu S H,Jiang W,et al.Characterization and mapping of a shattering mutant in rice that corresponds to a block of domestication genes.Genetics,2006,173(2):995-1005.
[12]Yoon J,Cho L H,Antt H W,et al.KNOX protein OSH15induces grain shattering by repressing lignin biosynthesis genes.Plant Physiology,2017,174(1):312-325.
[13]Zhao X H,Jiang X,Zhao K,et al.Screening of germplasm with low seed shattering rate and evaluation on agronomic traits in Elymus sibiricus L.Journal of Plant Genetic Resources,2015,16(4):691-699.赵旭红,姜旭,赵凯,等.低落粒老芒麦种质筛选及农艺性状综合评价.植物遗传资源学报,2015,16(4):691-699.
[14]Liu Z J,Chen Y,Meng J,et al.Seed shattering and relevant traits of Leymus chinensis.Acta Agrestia Sinica,2013,21(1):152-158.刘祝江,陈延,蒙静,等.羊草种子的落粒性及其相关特征的研究.草地学报,2013,21(1):152-158.
[15]Tian H,Zhang H S,Xiong J B,et al.Study on the seed shattering and suitable seed harvesting time of Bromus cartharticus Vahl.Hubei Agricultural Sciences,2015,54(23):5969-5972.田宏,张鹤山,熊军波,等.扁穗雀麦种子落粒性和适宜采收期的研究.湖北农业科学,2015,54(23):5969-5972.
[16]Bonin S G,Goplen B P.A histological study of seed shattering in reed canary grass.Canadian Journal of Plant Science,1963,43(2):200-205.
[17]Liu W H,Liang G L,Zhou Q P,et al.Study on shattering and growth physiological characteristics of Poa pratensis var.anceps during the process of seed development.Seed,2009,28(6):18-23.刘文辉,梁国玲,周青平,等.青海扁茎早熟禾种子成熟过程中落粒性与生长生理特性的研究.种子,2009,28(6):18-23.
[18]Elgersma A,Leeuwangh J E,Wilms H J.Abscission and seed shattering in perennial ryegrass(Lolium perenne L.).Euphytica,1988,39(3):51-57.
[19]Li W,Gill B S.Multiple genetic pathways for seed shattering in the grasses.Functional and Integrative Genomics,2006,6(4):300-309.
[20]Xie W,Zhang J,Zhao X,et al.Transcriptome profiling of Elymus sibiricus,an important forage grass in Qinghai-Tibet plateau,reveals novel insights into candidate genes that potentially connected to seed shattering.BMC Plant Biology,2017,17(1):78-92.
[21]Wang B D.Introduction and domestication in wild Elymus sibiricus.Chinese Journal of Grassland,1986,6(6):35-37.王比德.野生老芒麦的引种驯化.中国草地学报,1986,6(6):35-37.
[22]You M H,Liu J P,Bai S Q,et al.Study on relationship of seed shattering,seed development and yield traits of Elymus sibiricus L.Southwest China Journal of Agricultural Sciences,2011,24(4):1256-1260.游明鸿,刘金平,白史且,等.老芒麦落粒性与种子发育及产量性状关系的研究.西南农业学报,2011,24(4):1256-1260.
[23]Zhang Z Y,Zhang J C,Zhao X H,et al.Assessing and broadening genetic diversity of Elymus sibiricus germplasm for the improvement of seed shattering.Molecules,2016,21(7):869-882.
[24]Zhao X H,Xie W G,Zhang J C,et al.Histological characteristics,cell wall hydrolytic enzymes activity and candidate genes expression associated with seed shattering of Elymus sibiricus accessions.Frontiers in Plant Science,2017,8:1-13.
[25]Wang L Q,Yang J.Anatomical study on seed shattering mechanism of Poaceae grass.Inner Mongolia Prataculture,1995,(Z2):47-50.王立群,杨静.禾本科牧草种子脱落机制的解剖学研究.内蒙古草业,1995,(Z2):47-50.
[26]Zhang J C,Zhao X H,Zhang Z Y,et al.Seed shattering dynamic,and its relationship with agronomic traits and geographical distribution of Elymus sibiricus in Qinghai-Tibet Plateau.Acta Botanica Boreali-Occidentalia Sinica,2017,37(8):1595-1602.张俊超,赵旭红,张宗瑜,等.青藏高原老芒麦落粒动态与农艺性状及地理分布关系研究.西北植物学报,2017,37(8):1595-1602.
[27]Burson B L,Correa J,Potts H C.Anatomical basis for seed shattering in Kleingrass and Guineagrass.Cropence,1983,23(4):747-751.
[28]Ji H,Kim S R,Kim Y H,et al.Inactivation of the CTD phosphatase-like gene OsCPL1enhances the development of the abscission layer and seed shattering in rice.Plant Journal for Cell and Molecular Biology,2010,61(1):96-106.
[29]Yoon J,Cho L H,Kim S L,et al.The BEL1-type homeobox gene SH5induces seed shattering by enhancing abscission-zone development and inhibiting lignin biosynthesis.Plant Journal for Cell and Molecular Biology,2014,79(5):717-728.
[30]Thurber C S,Hepler P K,Caicedo A L.Timing is everything:early degradation of abscission layer is associated with increased seed shattering in U.S.weedy rice.BMC Plant Biology,2011,11(1):14.
[31]Inoue C,Htun T M,Inoue K,et al.Inhibition of abscission layer formation by an interaction of two seed-shattering loci,sh4and qSH3,in rice.Genes and Genetic Systems,2015,90(1):1-9.
[32]Zhang S,Zhang G L,Zeng X L.Advances in research on the effects of polygalacturonase and cellulase on fruit ripening.Journal of Fruit Science,2005,22(5):532-536.张嵩,张光伦,曾秀丽.纤维素酶和多聚半乳糖醛酸酶与果实成熟.果树学报,2005,22(5):532-536.
[33]Bonghi C,Rascio N,Ramina A,et al.Cellulase and polygalacturonase involvement in the abscission of leaf and fruit explants of peach.Plant Molecular Biology,1992,20(5):839.
[34]Greenberg J,Goren R,Riov J.The role of cellulase and polygalacturonase in abscission of young and mature shamouti orange fruits.Physiologia Plantarum,1975,34(1):1-7.
[35]Chen F H,Wu G B,Feng Z S,et al.The relation of grape berry abscission to changes of enzyme activity in abscission zone and plant growth regulators during storage.Journal of Plant Physiology and Molecular Biology,2003,29(2):133-140.陈发河,吴光斌,冯作山,等.葡萄贮藏过程中落粒与离区酶活性变化及植物生长调节物质的关系.植物生理与分子生物学学报,2003,29(2):133-140.
[36]Agrawal A P,Basarkar P W,Salimath P M,et al.Role of cell wall-degrading enzymes in pod-shattering process of soybean,Glycine max(L.)Merrill.Current Science,2002,82(1):58-61.
[2]Chen T,Nan Z B.Seed-borne fungi infection of Siberian wildrye:effects on seed germination and seedling growth.Acta Prataculturae Sinica,2015,24(2):96-103.陈焘,南志标.不同储存年限老芒麦种子种带真菌检测及致病性测定.草业学报,2015,24(2):96-103.
[3]Gu X Y,Guo Z H,Zhang X Q,et al.Genetic diversity of Elymus sibiricus germplasm resources revealed by SRAP markers.Acta Prataculturae Sinica,2014,23(1):205-216.顾晓燕,郭志慧,张新全,等.老芒麦种质资源遗传多样性的SRAP分析.草业学报,2014,23(1):205-216.
[4]Xie W,Zhao X,Zhang J,et al.Assessment of genetic diversity of Siberian wild rye(Elymus sibiricus L.)germplasms with variation of seed shattering and implication for future genetic improvement.Biochemical Systematics and Ecology,2015,58:211-218.
[5]Yan J J,Bai S Q,Ma X,et al.Genetic diversity of Elymus sibiricus and its breeding in China.Chinese Bulletin of Botany,2007,24(2):226-231.鄢家俊,白史且,马啸,等.老芒麦遗传多样性及育种研究进展.植物学报,2007,24(2):226-231.
[6]Konishi S,Izawa T,Lin S Y,et al.An SNP caused loss of seed shattering during rice domestication.Science,2006,312:1392-1396.
[7]Lin Z W,Li X R,Shannon L M,et al.Parallel domestication of the Shattering1genes in cereals.Nature Genetics,2012,44(6):720-724.
[8]Simons K J,Fellers J P,Trick H N,et al.Molecular characterization of the major wheat domestication gene Q.Genetics,2006,172(1):547-555.
[9]Kandemir N,Kudrna D A,Ullrich S E,et al.Molecular marker assisted genetic analysis of head shattering in six-rowed barley.Theoretical and Applied Genetics,2000,101(1):203-210.
[10]Pourkheirandish M,Hensel G,Kilian B,et al.Evolution of the grain dispersal system in barley.Cell,2015,162(3):527-539.
[11]Ji H S,Chu S H,Jiang W,et al.Characterization and mapping of a shattering mutant in rice that corresponds to a block of domestication genes.Genetics,2006,173(2):995-1005.
[12]Yoon J,Cho L H,Antt H W,et al.KNOX protein OSH15induces grain shattering by repressing lignin biosynthesis genes.Plant Physiology,2017,174(1):312-325.
[13]Zhao X H,Jiang X,Zhao K,et al.Screening of germplasm with low seed shattering rate and evaluation on agronomic traits in Elymus sibiricus L.Journal of Plant Genetic Resources,2015,16(4):691-699.赵旭红,姜旭,赵凯,等.低落粒老芒麦种质筛选及农艺性状综合评价.植物遗传资源学报,2015,16(4):691-699.
[14]Liu Z J,Chen Y,Meng J,et al.Seed shattering and relevant traits of Leymus chinensis.Acta Agrestia Sinica,2013,21(1):152-158.刘祝江,陈延,蒙静,等.羊草种子的落粒性及其相关特征的研究.草地学报,2013,21(1):152-158.
[15]Tian H,Zhang H S,Xiong J B,et al.Study on the seed shattering and suitable seed harvesting time of Bromus cartharticus Vahl.Hubei Agricultural Sciences,2015,54(23):5969-5972.田宏,张鹤山,熊军波,等.扁穗雀麦种子落粒性和适宜采收期的研究.湖北农业科学,2015,54(23):5969-5972.
[16]Bonin S G,Goplen B P.A histological study of seed shattering in reed canary grass.Canadian Journal of Plant Science,1963,43(2):200-205.
[17]Liu W H,Liang G L,Zhou Q P,et al.Study on shattering and growth physiological characteristics of Poa pratensis var.anceps during the process of seed development.Seed,2009,28(6):18-23.刘文辉,梁国玲,周青平,等.青海扁茎早熟禾种子成熟过程中落粒性与生长生理特性的研究.种子,2009,28(6):18-23.
[18]Elgersma A,Leeuwangh J E,Wilms H J.Abscission and seed shattering in perennial ryegrass(Lolium perenne L.).Euphytica,1988,39(3):51-57.
[19]Li W,Gill B S.Multiple genetic pathways for seed shattering in the grasses.Functional and Integrative Genomics,2006,6(4):300-309.
[20]Xie W,Zhang J,Zhao X,et al.Transcriptome profiling of Elymus sibiricus,an important forage grass in Qinghai-Tibet plateau,reveals novel insights into candidate genes that potentially connected to seed shattering.BMC Plant Biology,2017,17(1):78-92.
[21]Wang B D.Introduction and domestication in wild Elymus sibiricus.Chinese Journal of Grassland,1986,6(6):35-37.王比德.野生老芒麦的引种驯化.中国草地学报,1986,6(6):35-37.
[22]You M H,Liu J P,Bai S Q,et al.Study on relationship of seed shattering,seed development and yield traits of Elymus sibiricus L.Southwest China Journal of Agricultural Sciences,2011,24(4):1256-1260.游明鸿,刘金平,白史且,等.老芒麦落粒性与种子发育及产量性状关系的研究.西南农业学报,2011,24(4):1256-1260.
[23]Zhang Z Y,Zhang J C,Zhao X H,et al.Assessing and broadening genetic diversity of Elymus sibiricus germplasm for the improvement of seed shattering.Molecules,2016,21(7):869-882.
[24]Zhao X H,Xie W G,Zhang J C,et al.Histological characteristics,cell wall hydrolytic enzymes activity and candidate genes expression associated with seed shattering of Elymus sibiricus accessions.Frontiers in Plant Science,2017,8:1-13.
[25]Wang L Q,Yang J.Anatomical study on seed shattering mechanism of Poaceae grass.Inner Mongolia Prataculture,1995,(Z2):47-50.王立群,杨静.禾本科牧草种子脱落机制的解剖学研究.内蒙古草业,1995,(Z2):47-50.
[26]Zhang J C,Zhao X H,Zhang Z Y,et al.Seed shattering dynamic,and its relationship with agronomic traits and geographical distribution of Elymus sibiricus in Qinghai-Tibet Plateau.Acta Botanica Boreali-Occidentalia Sinica,2017,37(8):1595-1602.张俊超,赵旭红,张宗瑜,等.青藏高原老芒麦落粒动态与农艺性状及地理分布关系研究.西北植物学报,2017,37(8):1595-1602.
[27]Burson B L,Correa J,Potts H C.Anatomical basis for seed shattering in Kleingrass and Guineagrass.Cropence,1983,23(4):747-751.
[28]Ji H,Kim S R,Kim Y H,et al.Inactivation of the CTD phosphatase-like gene OsCPL1enhances the development of the abscission layer and seed shattering in rice.Plant Journal for Cell and Molecular Biology,2010,61(1):96-106.
[29]Yoon J,Cho L H,Kim S L,et al.The BEL1-type homeobox gene SH5induces seed shattering by enhancing abscission-zone development and inhibiting lignin biosynthesis.Plant Journal for Cell and Molecular Biology,2014,79(5):717-728.
[30]Thurber C S,Hepler P K,Caicedo A L.Timing is everything:early degradation of abscission layer is associated with increased seed shattering in U.S.weedy rice.BMC Plant Biology,2011,11(1):14.
[31]Inoue C,Htun T M,Inoue K,et al.Inhibition of abscission layer formation by an interaction of two seed-shattering loci,sh4and qSH3,in rice.Genes and Genetic Systems,2015,90(1):1-9.
[32]Zhang S,Zhang G L,Zeng X L.Advances in research on the effects of polygalacturonase and cellulase on fruit ripening.Journal of Fruit Science,2005,22(5):532-536.张嵩,张光伦,曾秀丽.纤维素酶和多聚半乳糖醛酸酶与果实成熟.果树学报,2005,22(5):532-536.
[33]Bonghi C,Rascio N,Ramina A,et al.Cellulase and polygalacturonase involvement in the abscission of leaf and fruit explants of peach.Plant Molecular Biology,1992,20(5):839.
[34]Greenberg J,Goren R,Riov J.The role of cellulase and polygalacturonase in abscission of young and mature shamouti orange fruits.Physiologia Plantarum,1975,34(1):1-7.
[35]Chen F H,Wu G B,Feng Z S,et al.The relation of grape berry abscission to changes of enzyme activity in abscission zone and plant growth regulators during storage.Journal of Plant Physiology and Molecular Biology,2003,29(2):133-140.陈发河,吴光斌,冯作山,等.葡萄贮藏过程中落粒与离区酶活性变化及植物生长调节物质的关系.植物生理与分子生物学学报,2003,29(2):133-140.
[36]Agrawal A P,Basarkar P W,Salimath P M,et al.Role of cell wall-degrading enzymes in pod-shattering process of soybean,Glycine max(L.)Merrill.Current Science,2002,82(1):58-61.