Effective RNAi-Mediated Silencing of the <i>Mismatch Repair MSH2</i> Gene Induces Sterility of Tomato Plants but Not an Increase in Meiotic Recombination

In plant breeding, the ability to manipulate meiotic recombination aids in the efficient construction of new allelic compositions of chromosomes and facilitates gene transfer from wild relatives of crop plants. The DNA mismatch repair system antagonizes meiotic recombination. In this research, a tri...

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Bibliographic Details
Main Authors: Svetlana R. Strelnikova, Anastasiya A. Krinitsina, Roman A. Komakhin
Format: Article
Language:English
Published: MDPI AG 2021-07-01
Series:Genes
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Online Access:https://www.mdpi.com/2073-4425/12/8/1167
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Summary:In plant breeding, the ability to manipulate meiotic recombination aids in the efficient construction of new allelic compositions of chromosomes and facilitates gene transfer from wild relatives of crop plants. The DNA mismatch repair system antagonizes meiotic recombination. In this research, a trial was conducted to evaluate transgenic tomato plants carrying an RNA interference (RNAi) construct designed to inhibit the expression of the mismatch repair <i>MSH2</i> gene. To drive the RNAi construct, we used either a pro-SmAMP2 promoter from <i>Stellaria media</i> ANTIMICROBIAL PEPTIDE2 or a Cauliflower mosaic virus 35S promoter (CaMV35S). The results of real-time PCR showed that, with a 16 h light/8 h dark photoperiod, MSH2-RNAi tomato transgenic plants exhibited <i>MSH2</i> gene transcript contents ranging from 0% to 3% in the leaves, relative to untransformed controls. However, with this lighting mode, the MSH2-RNAi transgenic plants grew slowly, flowered poorly, and did not form seed sets. During cultivation with a 12 h light/12 h dark photoperiod, MSH2-RNAi transgenic plants exhibited <i>MSH2</i> gene transcript contents ranging from 3% to 42%, relative to untransformed controls. Under these conditions, F<sub>1</sub> hybrid seed sets formed for most of the MSH2-RNAi transgenic plants with the RNAi construct driven by the CaMV35S promoter, and for one transformant with the RNAi construct driven by the pro-SmAMP2 promoter. Under conditions of a 12 h light/12 h dark photoperiod, most of the F<sub>1</sub> transgenic hybrids showed <i>MSH2</i> gene transcript contents ranging from 3% to 34% and formed F<sub>2</sub> offspring sets, which made it possible to assess the meiotic recombination frequency. We showed that the effective inhibition of <i>MSH2</i> in MSH2-RNAi tomato transgenic plants is not associated with an increase in meiotic recombination compared to the control, but it stimulates the sterility of plants. It was established that the expression of the <i>MSH2</i> gene in tomato plants is about 50 times higher with a 12 h light/12 h dark than with a 16 h light/8 h dark photoperiod. It is discussed that, in <i>Solanum lycopersicum</i> tomato plants, which are not sensitive to the day length for flowering, changing the lighting time may be a means of controlling the meiotic recombination frequency within certain limits.
ISSN:2073-4425