Reciprocal chromosome translocation associated with TDNA-insertion mutation in Arabidopsis: genetic and cytological analyses of consequences for gametophyte development and for construction of doubly mutant lines.
Reciprocal chromosome translocation associated with TDNA-insertion mutation in Arabidopsis: genetic and cytological analyses of consequences for gametophyte development and for construction of doubly mutant lines.
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DOI：
10.1007/s00425-008-0868-0
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发表时间：
2009-03
期刊：
影响因子：
4.3
通讯作者：
Hays JB
中科院分区：
文献类型：
--
作者：
Curtis MJ;Belcram K;Bollmann SR;Tominey CM;Hoffman PD;Mercier R;Hays JB
染色体重排可能使具有多个TDNA插入突变的拟南芥的构建复杂化。在这里，杂交两个纯合的AtREV 3和AtPOLH（染色体I和V，分别）插入和自交F1植株产生非孟德尔F2基因型分布：+/++/+和1/1 2/2后代的频率仅为0.42和0.25%。然而，正常发育和生育的双突变体显示AtPOLH-1和AtREV 3 -2配子和1/1 2/2胚胎是完全可行的。F2的分布可以定量预测，假设F1自交产生的（1，2）和（+，+）配子体的86%的时间。因此，F1自交过程本身的某些内在缺陷似乎是原因。在自交AtREV 3 +/2单突变体，胚珠和花粉成像显示逮捕或流产，分别有一半的配子体，然而，配子发生是正常的AtREV 3 2/2纯合子。这些发现，综合起来，表明T-DNA插入在AtREV 3的染色体上，I引起了相互I-V易位。在自交AtREV 3 +/2杂合子减数分裂I染色体的传播揭示了预测的十字形四染色体结构，荧光原位杂交显示总是包括易位和正常的染色体I和V。T-DNA与AtREV 3 DNA和两个与基因At 5g 59920的两个路口的测序建议易位通过独立的反向重复T-DNA插入之间的同源重组。因此，当TDNA插入突变体之间的杂交产生异常的后代分布时，应考虑TDNA连锁易位。
Chromosomal rearrangements may complicate construction of Arabidopsis with multiple TDNA-insertion mutations. Here, crossing two lines homozygous for insertions in AtREV3 and AtPOLH (chromosomes I and V, respectively) and selfing F1 plants yielded non-Mendelian F2 genotype distributions: frequencies of +/++/+ and 1/1 2/2 progeny were only 0.42 and 0.25%. However, the normal development and fertility of double mutants showed AtPOLH-1 and AtREV3-2 gametes and 1/1 2/2 embryos to be fully viable. F2 distributions could be quantitatively predicted by assuming that F1 selfing produced inviable (1,2) and (+,+) gametophytes 86% of the time. Some defect intrinsic to the F1 selfing process itself thus appeared responsible. In selfing AtREV3 +/2 single mutants, imaging of ovules and pollen showed arrest or abortion, respectively, of half of gametophytes; however, gametogenesis was normal in AtREV3 2/2 homozygotes. These findings, taken together, suggested that T-DNA insertion at AtREV3 on chromosome I had caused a reciprocal I–V translocation. Spreads of meiosis I chromosomes in selfing AtREV3 +/2 heterozygotes revealed the predicted cruciform four-chromosome structures, which fluorescence in situ hybridization showed to invariably include both translocated and normal chromosomes I and V. Sequencing of the two junctions of T-DNA with AtREV3 DNA and the two with gene At5g59920 suggested translocation via homologous recombination between independent inverted-repeat T-DNA insertions. Thus, when crosses between TDNA-insertion mutants yield anomalous progeny distributions, TDNA-linked translocations should be considered.
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3.2
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