Meiotic crossover reduction by virus-induced gene silencing enables the efficient generation of chromosome substitution lines and reverse breeding in Arabidopsis thaliana.,The Plant Journal

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Meiotic crossover reduction by virus-induced gene silencing enables the efficient generation of chromosome substitution lines and reverse breeding in Arabidopsis thaliana.
The Plant Journal ( IF 6.2 ) Pub Date : 2020-09-21 , DOI: 10.1111/tpj.14990
Vanesa Calvo-Baltanás ₁ , Cris L Wijnen ₁ , Chao Yang _{1,

2} , Nina Lukhovitskaya _{1,

3} , C Bastiaan de Snoo _{1,

4} , Linus Hohenwarter _{1,

2} , Joost J B Keurentjes ₁ , Hans de Jong ₁ , Arp Schnittger _{1,

2} , Erik Wijnker ₁

Affiliation

Plant breeding applications exploiting meiotic mutant phenotypes (like the increase or decrease of crossover (CO) recombination) have been proposed over the last years. As recessive meiotic mutations in breeding lines may affect fertility or have other pleiotropic effects, transient silencing techniques may be preferred. Reverse breeding is a breeding technique that would benefit from the transient downregulation of CO formation. The technique is essentially the opposite of plant hybridization: a method to extract parental lines from a hybrid. The method can also be used to efficiently generate chromosome substitution lines (CSLs). For successful reverse breeding, the two homologous chromosome sets of a heterozygous plant must be divided over two haploid complements, which can be achieved by the suppression of meiotic CO recombination and the subsequent production of doubled haploid plants. Here we show the feasibility of transiently reducing CO formation using virus‐induced gene silencing (VIGS) by targeting the meiotic gene MSH5 in a wild‐type heterozygote of Arabidopsis thaliana. The application of VIGS (rather than using lengthy stable transformation) generates transgene‐free offspring with the desired genetic composition: we obtained parental lines from a wild‐type heterozygous F₁ in two generations. In addition, we obtained 20 (of the 32 possible) CSLs in one experiment. Our results demonstrate that meiosis can be modulated at will in A. thaliana to generate CSLs and parental lines rapidly for hybrid breeding. Furthermore, we illustrate how the modification of meiosis using VIGS can open routes to develop efficient plant breeding strategies.

中文翻译：

病毒诱导的基因沉默导致的减数分裂交叉减少使得拟南芥中染色体替换系和反向育种的有效生成成为可能。

近年来，人们提出了利用减数分裂突变表型（例如交叉（CO）重组的增加或减少）的植物育种应用。由于育种系中的隐性减数分裂突变可能会影响生育力或具有其他多效性效应，因此可能优选瞬时沉默技术。反向育种是一种受益于二氧化碳形成瞬时下调的育种技术。该技术本质上与植物杂交相反：一种从杂种中提取亲本系的方法。该方法还可用于有效生成染色体替换系（CSL）。为了成功的反向育种，杂合植物的两个同源染色体组必须分为两个单倍体互补体，这可以通过抑制减数分裂CO重组和随后产生双单倍体植物来实现。在这里，我们展示了通过靶向拟南芥野生型杂合子中的减数分裂基因MSH5 ，使用病毒诱导的基因沉默（VIGS）暂时减少 CO 形成的可行性。 VIGS 的应用（而不是使用漫长的稳定转化）产生具有所需遗传组成的无转基因后代：我们从野生型杂合 F ₁中获得了两代亲本系。此外，我们在一项实验中获得了 20 个（32 个可能的）CSL。我们的结果表明，拟南芥减数分裂可以随意调节，以快速产生 CSL 和亲本系，用于杂交育种。此外，我们还说明了如何使用 VIGS 修饰减数分裂来开辟开发有效植物育种策略的途径。

更新日期：2020-09-21

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