QUARTET突变四倍体拟南芥的获得与表型分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0238

摘要/Abstract

摘要： 为了将四分子分析应用于多倍体遗传重组的研究,本研究从qrt1基因突变的二倍体Columbia（Col）与Landsberg erecta（Ler）生态型拟南芥出发,利用秋水仙素诱导核基因组加倍,利用流式细胞技术及荧光原位杂交技术筛选出成功加倍的qrt1基因突变的四倍体Col与Ler生态型拟南芥。并对其进行营养及生殖相关表型分析。首先qrt1突变抑制了侧根数量、基部侧枝数量以及莲座叶片面积。其次,四倍体莲座叶片面积、花粉、花药以及种子更大,侧根及茎基部分枝减少,抽薹时间推迟,每株果荚数与每果荚种子数更少,但种子发芽率都接近100%。倍性还对某些表型的影响具有背景依赖性,如对主茎分支数量、最终株高以及果荚长度的影响在Col与Ler生态型中表现不同。

关键词: 拟南芥, Quartet1, 四分子分析, 流式细胞技术, 荧光原位杂交, 表型分析

Abstract: To apply tetrad analysis in the research on genetic recombination of polyploid,we treated the diploid Columbia（Col）from gene qrt1 mutation and Arabidopsis thaliana of Landsberg erecta（Ler）ecotype with colchicine for inducing the genome doubling. The flow cytometry and fluorescence in situ hybridization were employed to screen the successfully-doubled tetraploid Col and Ler from gene qrt1 mutation. Then their vegetable and reproductive phenotypes were analyzed. As results,firstly,qrt1 mutation reduced the number of lateral roots and side bolts and the area of rosette leaves. Secondly,the area of rosette leaves and the size of pollen,anther and seed of the tetraploid increased,and the number of lateral roots and side bolts decreased,and the time of bolting siliques delayed,And the number of fruit pods per plant and seeds per fruit pod were less,while the germination rate approached 100%. Thirdly,some phenotypes were influenced differently by ploidy in Col and Ler,for example,the effects of ploidy on the height of stem,the number of branches and length of siliques varied.

Key words: Arabidopsis thaliana, Quartet1, tetrad analysis, flow cytometry, fluorescence in situ hybridization, phenotypic analysis

何硕康, 罗泽伟. QUARTET突变四倍体拟南芥的获得与表型分析[J]. 生物技术通报, 2018, 34(7): 119-125.

HE Shuo-kang, LUO Ze-wei. Development and Phenotypic Analysis of Tetraploid Arabidopsis thaliana with QUARTET Mutation[J]. Biotechnology Bulletin, 2018, 34(7): 119-125.

参考文献

[1] Leggatt RA, Iwama GK.Occurrence of polyploidy in the fishes[J]. Reviews in Fish Biology and Fisheries, 2003, 13(3):237-246.
[2] Lecher P, Defaye D, Noel P.Chromosomes and nuclear DNA of Crustacea[J]. Invertebrate Reproduction & Development, 1995, 27:85-114.
[3] Bachmann K, Bogart JP.Comparative cytochemical measurements in the diploid-tetraploid species pair of hylid frogs Hyla chrysoscelis and H. versicolor[J]. Cytogenet Cell Genet, 1975, 15(3):186-194.
[4] Otto SP, Whitton J.Polyploid incidence and evolution[J]. Annu Rev Genet, 2000, 34:401-437.
[5] Riddle NC, Birchler JA.Effects of reunited diverged regulatory hierarchies in allopolyploids and species hybrids[J]. Trends Genet, 2003, 19(11):597-600.
[6] Rausch JH, Morgan MT.The effect of self-fertilization, inbreeding depression, and population size on autopolyploid establishment[J]. Evolution, 2005, 59(9):1867-1875.
[7] Otto SP.The evolutionary consequences of polyploidy[J]. Cell, 2007, 131(3):452-462.
[8] Lloyd A, Bomblies K.Meiosis in autopolyploid and allopolyploid Arabidopsis[J]. Curr Opin Plant Biol, 2016, 30:116-122.
[9] NTJ B.Introduction to Mathematical Theory of Genetic Linkage[M]. London:Oxford University Press, 1961.
[10] Luo ZW, Zhang Z, Leach L, et al.Constructing genetic linkage maps under a tetrasomic model[J]. Genetics, 2006, 172(4):2635-2645.
[11] Pecinka A, Fang W, Rehmsmeier M, et al.Polyploidization increases meiotic recombination frequency in Arabidopsis[J]. BMC Biol, 2011, 9:24.
[12] Wang L, Luo Z.Polyploidization increases meiotic recombination frequency in Arabidopsis:a close look at statistical modeling and data analysis[J]. BMC Biol, 2012, 10:30.
[13] Winge O, Laustsen O.On two types of spore germination, and on genetic segregations in Saccharomyces demonstrated through single spore cultures[J]. CR Trav Lab Carlsberg Ser Physiol, 1937, 24:263-315.
[14] Preuss D, Rhee SY, Davis RW.Tetrad analysis possible in Arabidopsis with mutation of the QUARTET(QRT)genes[J]. Science, 1994, 264(5164):1458-1460.
[15] Rhee SY, Osborne E, et al.Microspore separation in the quartet 3 mutants of Arabidopsis is impaired by a defect in a developmentally regulated polygalacturonase required for pollen mother cell wall degradation[J]. Plant Physiol, 2003, 133(3):1170-1180.
[16] Galbraith DW, Harkins KR, Maddox JM, et al.Rapid flow cytometric analysis of the cell cycle in intact plant tissues[J]. Science, 1983, 220(4601):1049-1051.
[17] Heslop-Harrison JS.Cytogenetic analysis of Arabidopsis[J]. Methods Mol Biol, 1998, 82:119-127.
[18] Riechmann J, Wellmer F.Flower Development[M]. New York:Humana Press, 2014.
[19] Galbraith DW, et al.Systemic Endopolyp-loidy in Arabidopsis thaliana[J]. Plant Physiol, 1991, 96(3):985-989.
[20] Lu P, Han X, Qi J, et al.Analysis of Arabidopsis genome-wide variations before and after meiosis and meiotic recombination by resequencing Landsberg erecta and all four products of a single meiosis[J]. Genome Research, 2012, 22(3):508-518.
[21] Lu P, Han X, Qi J, et al.Analysis of Arabidopsis genome-wide variations before and after meiosis and meiotic recombination by resequencing Landsberg erecta and all four products of a single meiosis[J]. Genome Research, 2012, 22(3):508-518.
[22] Francis KE, Lam SY, Copenhaver GP.Separation of Arabidopsis pollen tetrads is regulated by QUARTET1, a pectin methylesterase gene[J]. Plant Physiol, 2006,142(3):1004-1013.