欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2010, Vol. 36 ›› Issue (07): 1216-1220.doi: 10.3724/SP.J.1006.2010.01216

• 研究简报 • 上一篇    下一篇

利用荧光原位杂交技术分析新合成异源四倍体拟南芥

位芳,张改生*   

  1. 西北农林科技大学 / 陕西省作物杂种优势研究与利用重点实验室 / 小麦育种教育部工程研究中心，陕西杨凌 712100
  • 收稿日期:2009-10-26 修回日期:2010-04-21 出版日期:2010-07-12 网络出版日期:2010-05-20
  • 通讯作者: 张改生, E-mail: zhanggsh@public.xa.sn.cn
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2009AA101102),陕西省13115科技创新工程重大科技专项(2007ZDKG-02)和国家杨凌农业生物技术育种中心专项基金项目(99-1A)资助.

FISH Analysis of Resynthesized Allotetraploid Arabidopsis

LI Fang,ZHANG Gai-Sheng   

  1. Key Laboratory of Crop Heterosis of Shaanxi Province, Wheat Breeding Engineering Research Center, Ministry of Education, Northwest A&F University, Yangling 712100, China
  • Received:2009-10-26 Revised:2010-04-21 Published:2010-07-12 Published online:2010-05-20
  • Contact: ZHANG Gai-Sheng, E-mail: zhanggsh@public.xa.sn.cn

PDF

2045

被引次数

4

摘要：

在植物异源倍化育种中或者通过异源多倍化导入有利基因时，初级杂交后代异源多倍体减数分裂期间，避免部分同源染色体干扰，使同源染色体正常联会和配对以及正确分离是产生功能性雌雄配子的细胞学基础。本研究通过种间杂交技术，获得初级杂交后代异源四倍体拟南芥A. suecica，并重点分析其花粉母细胞减数分裂期同源染色体联会和配对情况。利用DAPI技术染色证明了花粉母细胞减数分裂期间核内染色体数目的正确性和均等分裂；而用荧光原位杂交技术进一步证明了核内染色体组的来源的正确性和同源染色体联会和配对的精准性。该结果证实新合成异源多倍体能进行正常的减数分裂和产生功能性雌雄配子，为种属间杂交和异源多倍体育种以及植物杂种优势利用提供了有利的细胞遗传学证据。

关键词: 拟南芥, 新合成, 异源四倍体, DAPI, 荧光原位杂交

Abstract:

Allopolyploidy breeding or introduction of preferential gene(s) via allopolyploidization is widely attempted in production of new crops. Thus at the onset of establishment of new allopolyploids, the functional gametes are reproduced conditioning that normal events such as synapsis and pairing and correct segregation of homologous chromosomes are guaranteed with the avoidance of possible interference of homoeologous chromosomes during meiosis. In the present study, we reported on meiotic synapsis and pairing of homologous chromosomes during the development of pollen mother cells in the newly resynthesized allotetraploid Arabidopsis aided with DAPI staining and FISH technique. The results showed that the correct number of nuclear chromosomes and balanced segregation were frequently observed with DAPI staining, and FISH analysis further provided the improved resolution of synapsis and pairing of homologous chromosomes, and the investigated nuclear chromosomes derived from different parental lines had no interference with each other, and the synapsis and pairing of homologous chromosomes were clearly identified. Therefore, the results indicated that the newly resynthesized allotetraploid Arabidopsis may achieve the normal meiosis and propagate the functionalized gametes for fertilization, confirming a potential vigor of intra/interspecific hybridization and a cytological basis for allopolyploid breeding.

Key words: Arabidopsis, Allotertraploid, Resynthesized, DAPI, FISH


[1] Levsky J M, Singer R H. Fluorescence in situ hybridization: past, present and future. J Cell Sci, 2003, 116: 2833–2838


[2] Jiang J M, Gill B S. Current status and the future of fluorescence in situ hybridization (FISH) in plant genome research. Genome, 2006, 49: 1057–1068


[3] Gorman P, Roylance R. Fluorescence in situ hybridization and comparative genomic hybridization. Methods Mol Med, 2006, 120: 269–295


[4] Ali H B, Lysak M A, Schubert I. Genomic in situ hybridization in plants with small genomes is feasible and elucidates the chromosomal parentage in interspecific Arabidopsis hybrids. Genome, 2004, 47: 954–960


[5] Doyle J J, Flagel L E, Paterson A H, Rapp R A, Soltis D E, Soltis P S, Wendel J F. Evolutionary genetics of genome merger and doubling in plants. Annu Rev Genet, 2008, 42: 443–461


[6] Comai L, Tyagi A P, Winter K, Holmes-Davis R, Reynolds S H, Stevens Y, Byers B. Phenotypic instability and rapid gene silencing in newly formed Arabidopsis allotetraploids. Plant Cell, 2000, 12: 1551–1568


[7] Schranz M E, Osborn T C. Novel flowering time variation in the resynthesized polyploid Brassica napus. J Hered, 2000, 91: 242–246


[8] Madlung A, Tyagi A P, Watson B, Jiang H, Kagochi T, Doerge R W, Martienssen R, Comai L. Genomic changes in synthetic Arabidopsis polyploids. Plant J, 2005, 41: 221–230


[9] Song K, Lu P, Tang K, Osborn T C. Rapid genome change in synthetic polyploids of Brassica and its implications for polyploid evolution. Proc Natl Acad Sci USA, 1995, 92: 7719–7723


[10] Pikaard C S. Genomic change and gene silencing in polyploids. Trends Genet, 2001, 17: 675–677


[11] Chen Z J. Genetic and epigenetic mechanisms for gene expression and phenotypic variation in plant polyploids. Annu Rev Plant Biol, 2007, 58: 377–406


[12] Liu B, Wendel J F. Epigenetic phenomena and the evolution of plant allopolyploids. Mol Phylogenet Evol, 2003, 29: 365–379


[13] Roberts M A, Reader S M, Dalgliesh C, Miller T E, Foote T N, Fish L J, Snape J W, Moore G. Induction and characterization of Ph1 wheat mutants. Genetics, 1999, 153: 1909–1918


[14] Griffiths S, Sharp R, Foote T N, Bertin I, Wanous M, Reader S, Colas I, Moore G. Molecular characterization of Ph1 as a major chromosome pairing locus in polyploid wheat. Nature, 2006, 439: 749–752


[15] Comai L, Madlung A, Josefsson C, Tyagi A. Do the different parental 'heteromes' cause genomic shock in newly formed allopolyploids. Philos Trans R Soc Lond B Biol Sci, 2003, 358: 1149–1155


[16] Wang A-Y(王爱云), Chen D-L(陈冬玲), Cai D-T(蔡得田). Applications of wide hybridization and allopolyploidization in Rice Breeding. J Wuhan Bot Res (武汉植物学研究), 2005, 23(5) : 491–4954 (in Chinese with English abstract)


[17] Stewart C N Jr, Via L E. A rapid CTAB DNA isolation technique useful for RAPD fingerprinting and other PCR applications. Biotechniques, 1993, 14: 748–50


[18] Ross K J, Fransz P, Jones G H. A light microscopic atlas of meiosis in Arabidopsis thaliana. Chromosome Res, 1996, 4: 507–516


[19] Armstrong S J, Sanchez-Moran E, Franklin F C. Cytological analysis of Arabidopsis thaliana meiotic chromosomes. Methods Mol Biol, 2009, 558: 131–145


[20] Comai L. The advantages and disadvantages of being polyploid. Nat Rev Genet, 2005, 6: 836–846


[21] Zhuang Y(庄勇), Chen L-Z(陈龙正), Yang Y-G(杨寅桂), Lou Q-F(娄群峰), Chen J-F(陈劲枫). Changes in gene expression in evolution of plant allopolyploids. Chin Bull Bot (植物学通报), 2006, 23(2): 207–214 (in Chinese with English abstract)


[22] Xiong Z-Y(熊志勇), Gao Y(高原), He G-Y(何光源), Gu M-G(谷明光), Guo L-Q(郭乐群), Song Y-C(宋运淳). Distribution of the knob heterochromatin repeat sequence on chromosome in maize, perennial diploid maize and their offspring. Chin Sci Bull (科学通报), 2004, 12(49): 1162–1165 (in Chinese with English abstract)


[23] Wang J L, Tian L, Lee H S, Wei N E, Jiang H M, Brian W, Andreas M, Osborn T C, Doerge R W, Comai L, Chen Z J. Genomewide nonadditive gene regulation in Arabidopsis allotetraploids. Genetics, 2006, 172: 507–517


[24] Beaulieu J, Jean M, Belzile F. The allotetraploid Arabidopsis thaliana-Arabidopsis lyrata subsp. petraea as an alternative model system for the study of polyploidy in plants. Mol Genet Genom, 2009, 281: 421–435


[25] Moore G. Meiosis in allopolyploids—the importance of ‘Teflon’ chromosomes. Trends Genet, 2002, 18: 456–463
[1] 许静, 毕竞男, 殷祥贞, 赵健鑫, 赵旭红, 潘丽娟, 陈娜, 姜骁, 马俊卿, 殷冬梅, 迟晓元. 花生ω-3脂肪酸脱氢酶基因在拟南芥中异源表达及转录组分析[J]. 作物学报, 2025, 51(12): 3251-3265.
[2] 薛丽, 李心怡, 黄勇泰, 欧财篮, 吴小青, 余泽怀, 崔泽田, 张木清, 邓祖湖, 余凡. 甘蔗与斑茅杂交染色体组构成特征研究[J]. 作物学报, 2024, 50(3): 633-644.
[3] 陈天, 李昱樱, 荣二花, 吴玉香. 棉属人工异源四倍体后代性状鉴定及花器转录组学分析[J]. 作物学报, 2024, 50(2): 325-339.
[4] 付佳祺, 李世宽, 谭萌慧, 罗方, 张传玲, 刘祾悦, 卢倩, 谷勇哲. 大豆GmRSM1通过调节PIN基因表达促进顶端弯钩消失[J]. 作物学报, 2024, 50(11): 2731-2741.
[5] 朱旭东, 杨兰锋, 陈媛媛, 侯泽豪, 罗旖柔, 熊泽浩, 方正武. 甜荞FeSGT1基因克隆及抗旱功能解析[J]. 作物学报, 2023, 49(6): 1573-1583.
[6] 史培瑶, 陈丽娟, 孙昊杰, 程梦豪, 肖进, 袁春霞, 王秀娥, 王海燕. 顶芒山羊草特异寡核苷酸探针开发和oligo-FISH核型构建[J]. 作物学报, 2023, 49(6): 1455-1465.
[7] 孟颖, 邢蕾蕾, 曹晓红, 郭光艳, 柴建芳, 秘彩莉. 小麦Ta4CL1基因的克隆及其在促进转基因拟南芥生长和木质素沉积中的功能[J]. 作物学报, 2022, 48(1): 63-75.
[8] 鲁海琴, 陈丽, 陈磊, 张盈川, 文静, 易斌, 涂金星, 傅廷栋, 沈金雄. Bna-novel-miR311-HSC70-1模块调控甘蓝型油菜响应热胁迫的机制[J]. 作物学报, 2020, 46(10): 1474-1484.
[9] 田文刚,朱雪峰,宋雯,程文翰,薛飞,朱华国. 异源表达棉花S-腺苷甲硫氨酸脱羧酶(GhSAMDC1)基因提高了拟南芥抗盐能力[J]. 作物学报, 2019, 45(7): 1017-1028.
[10] 赵翔,朱自亿,王潇楠,慕世超,张骁. 拟南芥RPT2与RIP1互作调节下胚轴向光弯曲的功能鉴定[J]. 作物学报, 2018, 44(12): 1802-1808.
[11] 李丽娜,杜培,付留洋,刘华,徐静,秦利,严玫,韩锁义,黄冰艳,董文召. 花生栽培种与野生种(Arachis oteroi)人工杂交双二倍体的创制和鉴定[J]. 作物学报, 2017, 43(01): 133-140.
[12] 刘睿洋，刘芳，张振乾，官春云. 甘蓝型油菜BnFAD2-C5基因启动子及内含子在表达水平的功能分析[J]. 作物学报, 2016, 42(10): 1471-1478.
[13] 刘凌云,刘浩,赵晶,王艳霞,王棚涛. 拟南芥低叶绿素荧光LCF3基因的克隆与功能分析[J]. 作物学报, 2016, 42(05): 690-695.
[14] 宋仲戬,张登峰*,李永祥,石云素,宋燕春,王天宇,黎裕. 玉米分子伴侣基因ZmBiP2在逆境下的功能分析[J]. 作物学报, 2015, 41(05): 708-716.
[15] 赵青平，赵翔，慕世超，肖慧丽，张骁. 拟南芥下胚轴向光弯曲P2SA2基因的克隆与功能鉴定[J]. 作物学报, 2015, 41(04): 585-592.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2