洋葱黄色条纹突变体的特征特性
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摘要
对母系遗传洋葱叶片黄色条纹突变体材料进行田间表型特征观察,叶绿素含量测定和显微结构观察,并利用jnurf13分子标记鉴定保持株。结果表明,洋葱突变体为亮绿黄色条纹。黄皮、白皮和紫皮洋葱中均发现黄色条纹突变株,黄化程度越高植株长势越弱;整个生育期黄色条纹一直存在于管状叶、花薹、花苞、伞状花序中花梗、花瓣、雌蕊、雄蕊等部位,直至植株枯萎也不返绿。单株自交或开放式自然授粉F1中出现全黄株、黄绿条纹株和绿株3种类型,无固定分离比,全黄株苗期死亡。洋葱黄色条纹突变体黄色组织的总叶绿素、叶绿素a和叶绿素b含量显著低于绿色组织;石蜡切片观察结果显示黄色和绿色组织的形态结构无差异,但绿色组织的叶绿体总数高于黄色组织;通过超显微结构观察发现,黄色条纹突变体绿色组织中叶绿体结构与正常株无差异,而黄色组织中类囊体、基粒等结构降解,嗜饿颗粒数量多且集中,影响光合作用。另外,利用jnurf13分子标记从12株可育突变体中筛选出5株保持株,可作为形态学标记用于洋葱细胞质雄性杂交制种。
In this study,field phenotypic characteristics of maternally inherited onion leaf stripe mutants,chlorophyll content and microstructure were observed. In addition,jnurf13 molecular markers were used to identify the maintained strains.The results showed that the onion mutants were bright green-yellow stripes and were found in yellow,white and purple onions.The higher the degree of yellowing,the weaker the growth of the plants. Yellow stripes have been accompanied by tubular leaves,calyx,peduncles,petals,pistils,stamens,until withered and not green. Single-plant self-cross or open natural pollination F1 appeared in all three types of whole yellow,yellow stripes and green strains,no fixed separation ratio and the whole yellow plant died at seedling stage. The contents of total chlorophyll,chlorophyll a and chlorophyll b in yellow tissue of onion yellow stripe mutant were significantly lower than those in green tissue. The results of paraffin sections observation showed that there was no difference in morphological structure between green and yellow tissues,but the total number of chloroplasts in green tissues was higher than that in yellow tissues. Ultrastructural observation revealed that there was no difference in chloroplast structure between the green mutant tissue and the normal strain,whereas thylakoid and grana in yellow tissue were degraded,and the number of hungry particles was numerous and concentrated,affecting photosynthesis. In addition,five conserved strains were screened out from 12 fertile mutants using the jnurf13 molecular marker and used as a morphological marker for onion cytoplasmic male hybrid production.
In this study,field phenotypic characteristics of maternally inherited onion leaf stripe mutants,chlorophyll content and microstructure were observed. In addition,jnurf13 molecular markers were used to identify the maintained strains.The results showed that the onion mutants were bright green-yellow stripes and were found in yellow,white and purple onions.The higher the degree of yellowing,the weaker the growth of the plants. Yellow stripes have been accompanied by tubular leaves,calyx,peduncles,petals,pistils,stamens,until withered and not green. Single-plant self-cross or open natural pollination F1 appeared in all three types of whole yellow,yellow stripes and green strains,no fixed separation ratio and the whole yellow plant died at seedling stage. The contents of total chlorophyll,chlorophyll a and chlorophyll b in yellow tissue of onion yellow stripe mutant were significantly lower than those in green tissue. The results of paraffin sections observation showed that there was no difference in morphological structure between green and yellow tissues,but the total number of chloroplasts in green tissues was higher than that in yellow tissues. Ultrastructural observation revealed that there was no difference in chloroplast structure between the green mutant tissue and the normal strain,whereas thylakoid and grana in yellow tissue were degraded,and the number of hungry particles was numerous and concentrated,affecting photosynthesis. In addition,five conserved strains were screened out from 12 fertile mutants using the jnurf13 molecular marker and used as a morphological marker for onion cytoplasmic male hybrid production.
引文
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[12] TATEBE T. Genetic studies on the leaf variegation of Allium fistu-losum linnaeus[J]. The Japanese Journal of Genetics,1961,36:151-156.
[13] TATEBE T. Genetic studies on the leaf variegation of Allium cepaL.[J]. Journal of the Japanese Society for Horticultural Science,1968,37(4):345-348.
[14] KIM S. A codominant molecular marker in linkage disequilibriumwith a restorer-of-fertility gene(Ms)and its application in reevalu-ation of inheritance of fertility restoration in onions[J]. MolecularBreeding,2014,34:769-778.
[15]李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:134.
[16]何俊平,朱家成,王建平,等.甘蓝型油菜幼苗显微组织结构与抗寒性的关系[J].江苏农业学报,2017,33(1):19-26.
[17]曾爱松,宋立晓,高兵,等.低温胁迫下甘蓝叶片细胞超微结构及叶肉细胞内Ca2+的分布变化[J].江苏农业学报,2011,27(5):1053-1058.
[18] KOBAYASHI N,MATSUNAGA M,NAKATSUKA A,et al. Chi-meric inheritance of organelle DNA in variegated leaf seedlingsfrom inter-subgeneric crossing of azalea[J]. Euphytica,2013,191(1):121-128.
[19]常青山,张利霞,陈煜,等.菊花黄绿叶突变体的光合与类囊体膜光谱[J].林业科学,2013,49(2):72-78.
[20]李燕群,钟萍,高志艳,等.水稻斑马叶突变体zebra524的表型鉴定及候选基因分析[J/OL].中国农业科学,2014,47(15):2907-2915.
[21] KAZUO T,MASAHIKO M,KYOKO T,et al. An active DNAtransposon n Dart causing leaf variegation and mutable dwarfismand its related elements in rice[J]. The Plant Journal,2006,45(1):46-57.
[22]曹莉.一个新的小麦黄化突变体研究[D].杨凌:西北农林科技大学,2007.
[23] EFRON Y. An ems-sensitivity factor in maize conditioning albinoleaf stripes[J]. Genetics,1974,78(3):859-867.
[24] LI W X,YANG S B,LU Z G,et al. Cytological,physiological,and transcriptomic analyses of golden leaf coloration in Ginkgo bi-loba L.[J]. Horticulture Research,2018,5(1):12.
[25]王彩霞,田韦韦,田敏,等.文心兰黄化突变体的初步研究[J].核农学报,2013,27(12):1845-1852.
[26] LYU J,WANG Y H,LIU L L,et al. A putative plastidial adeninenucleotide transporter,BRITTLE1-3,plays an essential role inregulating chloroplast development in rice(Oryza sativa L.)[J].Journal of Plant Biology,2017,60(5):493-505.
[27] GE C W,WANG L,YE W J,et al. Single-point mutation of anhistidine-aspartic domain-containing gene involving in chloroplastribosome biogenesis leads to white fine stripe leaf in rice[J]. Sci-entific Reports,2017,7(1):3298.
[28]赵洪兵,郭会君,赵林姝,等.空间环境诱变小麦叶绿素缺失突变体的主要农艺性状和光合特性[J].作物学报,2011,37(1):119-126.
[29] CHEN X Q,ZHU L,XIN L,et al. Rice stripe1-2 and stripe1-3mutants encoding the small subunit of ribonucleotide reductase aretemperature sensitive and are required for chlorophyll biosynthesis[J]. PLoS ONE,2015,10(6):e0130172.
[30]齐新丽,徐智斌,裴洪翠,等.大麦EMS突变群体的创建及功能评价[J].麦类作物学报,2012,32(5):846-852.
[31] ZHAO S L,LONG W H,WANG W H,et al. A rice White-stripeleaf3(wsl3)mutant lacking an HD domain-containing protein af-fects chlorophyll biosynthesis and chloroplast development[J].Journal of Plant Biology,2016,59(3):282-292.
[32] XYA J,DENGA Y W,LIA Q,et al. STRIPE2 Encodes a putativedCMP deaminase that plays an important role in chloroplast devel-opment in rice[J]. Journal of Genetics and Genomics,2014,41(10):539-548.
[33] HE Y H,ZOU G X,RAO Y C,et al. Genetic analysis and genemapping of a rice white stripe leaf mutant(st10)[J]. Plant Geneand Trait,2011,2(4):23-29.
[34] WANG Y,REN Y,ZHOU K,et al. WHITE STRIPE LEAF4 en-codes a novel P-Type PPR protein required for chloroplast biogene-sis during early leaf development[J]. Frontiers of Plant Science,2017,8:1116.
[35] QIN R,ZENG D,LIANG R,et al. Rice gene SDL/RNRS1,en-coding the small subunit of ribonucleotide reductase,is requiredfor chlorophyll synthesis and plant growth development[J]. Gene,2017,627:351-362.
[36]成浩,陈明,虞富莲,等.茶叶片阶段性返白过程中色素蛋白复合体的变化[J].植物生理学通讯,2000,36(4):300-304.
[37] GU J,MILES D,Ne EWTON K J. Analysis of leaf sectors in theNCS6 mitochondrial mutant of maize[J]. The Plant Cell,1993,5(8):963-971.
[38] LI H C,QIAN Q,WANG Y,et al. Characterization and mappingof a white panicle mutant gene in rice[J]. Chinese Science Bulle-tin,2003,48(5):457-459.
[39]魏彦林,施勇烽,吴建利.水稻核基因控制的叶色变异[J].核农学报,2011,25(6):1169-1178.
[40] ZHAO F M,ZHANG Y,WU Y R,et al. Morphological and phys-iological analysis of narrow and striped leaf 1(nsl1)mutant of rice(Oryza sativa L.)and the gene mapping[J]. Chinese ScienceBulletin,2014,59(9):840-848.
[41]贾学静,冉何陈,曾顺华,等.园艺观赏植物金心吊兰的叶片解剖结构[J].四川农业大学学报,2011,29(2):199-202.
[42]常青山.菊花黄绿叶突变体黄叶与绿叶组织形成的生理与分子机制比较研究[D].南京:南京农业大学,2011.
[43]刘国民,高必军,文绍山.带有黄绿叶色标记的香型籼稻不育系形态特征及开花习性[J].分子植物育种,2009,7(5):916-921.
[44] WANG Q M,WANG L,ZHOU Y B,et al. Leaf patterning ofClivia miniata var. variegata is associated with differential DNAmethylation[J]. Plant Cell Reports,2016,35(1):167-184.
[2] ZHANG Z G,CUI X A,WANG Y W,et al. The RNA editingfactor WSP1 is essential for chloroplast development in rice[J].Molecular Plant(Cell Press),2017,10(1):86-98.
[3] QIN D D,DONG J,XU F C,et al. Characterization and finemapping of a novel barley stage green-revertible albino gene(Hv SGRA)by bulked segregant analysis based on SSR assay andspecific length amplified fragment sequencing[J]. BMC Genom-ics,2015,16:838.
[4] LIN B Y,YU H J. Inheritance of a striped-leaf mutant is associat-ed with the cytoplasmic genome in maize[J]. Theoretical and Ap-plied Genetics,1995,91:915-920.
[5]田韦韦,王彩霞,田敏,等.文心兰浅绿条纹突变体的生理生化及叶绿素荧光特性研究[J].西北植物学报,2015,35(10):2012-2017.
[6]陈天国.毛竹珍稀新栽培变种-绿槽龟甲竹[J].世界竹藤通讯,2014,12(2):23-24.
[7] YE W J,HU S K,WU L W,et al. White stripe leaf 12(WSL12),encoding a nucleoside diphosphate kinase 2(Os NDPK2),regu-lates chloroplast development and abiotic stress response in rice(Oryza sativa L.)[J]. Molecular Breeding,2016,36(5):57-72.
[8] REDDY M K,SUBRAHMANYAM N C,APPA RAO S,et al. Ul-trastructural and molecular characterization of altered plastids innuclear gene controlled yellow stripe mutant of Pennisetum ameri-canum[J]. Hereditas,1988,109(2):253-260.
[9] ZHAO H B,GUO H J,ZHAO L S,et al. Agronomic traits andphotosynthetic characteristics of chlorophyll-deficient wheat mutantinduced by spaceflight environment[J]. Acta Agronomica Sinica,2011,37(1):119-126.
[10] GU J,MILES D,NEWTON K J. Analysis of leaf sectors in theNCS6 mitochondrial mutant of maize[J]. The Plant Cell,1993,5(8):963-971.
[11] SUJATHA M,SUBRAHMANYAM N C. Characterisation of nucle-ar gene controlled yellow stripe mutant of Pennisetum glaucum(L.)R.Br.[J]. Plant Science,1991,73(1):55-64.
[12] TATEBE T. Genetic studies on the leaf variegation of Allium fistu-losum linnaeus[J]. The Japanese Journal of Genetics,1961,36:151-156.
[13] TATEBE T. Genetic studies on the leaf variegation of Allium cepaL.[J]. Journal of the Japanese Society for Horticultural Science,1968,37(4):345-348.
[14] KIM S. A codominant molecular marker in linkage disequilibriumwith a restorer-of-fertility gene(Ms)and its application in reevalu-ation of inheritance of fertility restoration in onions[J]. MolecularBreeding,2014,34:769-778.
[15]李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:134.
[16]何俊平,朱家成,王建平,等.甘蓝型油菜幼苗显微组织结构与抗寒性的关系[J].江苏农业学报,2017,33(1):19-26.
[17]曾爱松,宋立晓,高兵,等.低温胁迫下甘蓝叶片细胞超微结构及叶肉细胞内Ca2+的分布变化[J].江苏农业学报,2011,27(5):1053-1058.
[18] KOBAYASHI N,MATSUNAGA M,NAKATSUKA A,et al. Chi-meric inheritance of organelle DNA in variegated leaf seedlingsfrom inter-subgeneric crossing of azalea[J]. Euphytica,2013,191(1):121-128.
[19]常青山,张利霞,陈煜,等.菊花黄绿叶突变体的光合与类囊体膜光谱[J].林业科学,2013,49(2):72-78.
[20]李燕群,钟萍,高志艳,等.水稻斑马叶突变体zebra524的表型鉴定及候选基因分析[J/OL].中国农业科学,2014,47(15):2907-2915.
[21] KAZUO T,MASAHIKO M,KYOKO T,et al. An active DNAtransposon n Dart causing leaf variegation and mutable dwarfismand its related elements in rice[J]. The Plant Journal,2006,45(1):46-57.
[22]曹莉.一个新的小麦黄化突变体研究[D].杨凌:西北农林科技大学,2007.
[23] EFRON Y. An ems-sensitivity factor in maize conditioning albinoleaf stripes[J]. Genetics,1974,78(3):859-867.
[24] LI W X,YANG S B,LU Z G,et al. Cytological,physiological,and transcriptomic analyses of golden leaf coloration in Ginkgo bi-loba L.[J]. Horticulture Research,2018,5(1):12.
[25]王彩霞,田韦韦,田敏,等.文心兰黄化突变体的初步研究[J].核农学报,2013,27(12):1845-1852.
[26] LYU J,WANG Y H,LIU L L,et al. A putative plastidial adeninenucleotide transporter,BRITTLE1-3,plays an essential role inregulating chloroplast development in rice(Oryza sativa L.)[J].Journal of Plant Biology,2017,60(5):493-505.
[27] GE C W,WANG L,YE W J,et al. Single-point mutation of anhistidine-aspartic domain-containing gene involving in chloroplastribosome biogenesis leads to white fine stripe leaf in rice[J]. Sci-entific Reports,2017,7(1):3298.
[28]赵洪兵,郭会君,赵林姝,等.空间环境诱变小麦叶绿素缺失突变体的主要农艺性状和光合特性[J].作物学报,2011,37(1):119-126.
[29] CHEN X Q,ZHU L,XIN L,et al. Rice stripe1-2 and stripe1-3mutants encoding the small subunit of ribonucleotide reductase aretemperature sensitive and are required for chlorophyll biosynthesis[J]. PLoS ONE,2015,10(6):e0130172.
[30]齐新丽,徐智斌,裴洪翠,等.大麦EMS突变群体的创建及功能评价[J].麦类作物学报,2012,32(5):846-852.
[31] ZHAO S L,LONG W H,WANG W H,et al. A rice White-stripeleaf3(wsl3)mutant lacking an HD domain-containing protein af-fects chlorophyll biosynthesis and chloroplast development[J].Journal of Plant Biology,2016,59(3):282-292.
[32] XYA J,DENGA Y W,LIA Q,et al. STRIPE2 Encodes a putativedCMP deaminase that plays an important role in chloroplast devel-opment in rice[J]. Journal of Genetics and Genomics,2014,41(10):539-548.
[33] HE Y H,ZOU G X,RAO Y C,et al. Genetic analysis and genemapping of a rice white stripe leaf mutant(st10)[J]. Plant Geneand Trait,2011,2(4):23-29.
[34] WANG Y,REN Y,ZHOU K,et al. WHITE STRIPE LEAF4 en-codes a novel P-Type PPR protein required for chloroplast biogene-sis during early leaf development[J]. Frontiers of Plant Science,2017,8:1116.
[35] QIN R,ZENG D,LIANG R,et al. Rice gene SDL/RNRS1,en-coding the small subunit of ribonucleotide reductase,is requiredfor chlorophyll synthesis and plant growth development[J]. Gene,2017,627:351-362.
[36]成浩,陈明,虞富莲,等.茶叶片阶段性返白过程中色素蛋白复合体的变化[J].植物生理学通讯,2000,36(4):300-304.
[37] GU J,MILES D,Ne EWTON K J. Analysis of leaf sectors in theNCS6 mitochondrial mutant of maize[J]. The Plant Cell,1993,5(8):963-971.
[38] LI H C,QIAN Q,WANG Y,et al. Characterization and mappingof a white panicle mutant gene in rice[J]. Chinese Science Bulle-tin,2003,48(5):457-459.
[39]魏彦林,施勇烽,吴建利.水稻核基因控制的叶色变异[J].核农学报,2011,25(6):1169-1178.
[40] ZHAO F M,ZHANG Y,WU Y R,et al. Morphological and phys-iological analysis of narrow and striped leaf 1(nsl1)mutant of rice(Oryza sativa L.)and the gene mapping[J]. Chinese ScienceBulletin,2014,59(9):840-848.
[41]贾学静,冉何陈,曾顺华,等.园艺观赏植物金心吊兰的叶片解剖结构[J].四川农业大学学报,2011,29(2):199-202.
[42]常青山.菊花黄绿叶突变体黄叶与绿叶组织形成的生理与分子机制比较研究[D].南京:南京农业大学,2011.
[43]刘国民,高必军,文绍山.带有黄绿叶色标记的香型籼稻不育系形态特征及开花习性[J].分子植物育种,2009,7(5):916-921.
[44] WANG Q M,WANG L,ZHOU Y B,et al. Leaf patterning ofClivia miniata var. variegata is associated with differential DNAmethylation[J]. Plant Cell Reports,2016,35(1):167-184.