Promoters pro-SmAMP1 and pro-SmAMP2 from Wild Plant Stellaria media for the Biotechnology of Dicotyledons

Маджарова, Н. В.; Казакова, К. А.; Strelnikova, S. R.; Снычева, О. А.; Ветчинкина, Е. М.; Efremova, L. N.; Vysotskii, D. A.; Бабаков, А. В.; Комахин, Р. А.

doi:10.1134/s1021443718040040

Cited by 7 publications

(1 citation statement)

References 27 publications

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“…We found that the differences between these nucleotide sequences of the MSH2 gene transcripts from different species did not exceed 1%; therefore, even in interspecific tomato hybrids, a single construct based on the conserved region of the tomato MSH2 gene transcript can be used for silencing by RNAi. In addition, we previously developed a new gene promoter pro-SmAMP2 from Stellaria media ANTIMICROBIAL PEPTIDE2, which, in the leaves of transgenic tobacco plants (Nicotiana tabacum L.), is 2-3 times stronger than the viral promoter CaMV35S [22,23]. The pro-SmAMP2 promoter is also effective in the floral organs of transgenic tobacco plants, including the stem, sepals, pistil, stigma, anthers, pollen, and the microsporocytes during prophase I of meiosis [24] when crossing over occurs.…”

Section: Introductionmentioning

confidence: 99%

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

Strelnikova

Krinitsina

Комахин

2021

Genes

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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 MSH2 gene. To drive the RNAi construct, we used either a pro-SmAMP2 promoter from Stellaria media 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 MSH2 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 MSH2 gene transcript contents ranging from 3% to 42%, relative to untransformed controls. Under these conditions, F1 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 F1 transgenic hybrids showed MSH2 gene transcript contents ranging from 3% to 34% and formed F2 offspring sets, which made it possible to assess the meiotic recombination frequency. We showed that the effective inhibition of MSH2 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 MSH2 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 Solanum lycopersicum 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.

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Section: Introductionmentioning

confidence: 99%