All-Russia Research Institute of Agricultural Biotechnology, 127550 Moscow, Russia.
Biological Faculty, Lomonosov Moscow State University, 119234 Moscow, Russia.
Genes (Basel). 2021 Jul 29;12(8):1167. doi: 10.3390/genes12081167.
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 gene. To drive the RNAi construct, we used either a pro-SmAMP2 promoter from 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 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 gene transcript contents ranging from 3% to 42%, relative to untransformed controls. Under these conditions, F 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 transgenic hybrids showed gene transcript contents ranging from 3% to 34% and formed F offspring sets, which made it possible to assess the meiotic recombination frequency. We showed that the effective inhibition of 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 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 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.
在植物育种中，操纵减数分裂重组的能力有助于高效构建染色体的新等位基因组成，并促进作物野生亲缘植物的基因转移。DNA 错配修复系统拮抗减数分裂重组。在这项研究中，我们进行了一项试验，评估了携带设计用于抑制错配修复基因表达的 RNA 干扰 (RNAi) 构建体的转基因番茄植物。为了驱动 RNAi 构建体，我们使用了来自抗菌肽 2 的 pro-SmAMP2 启动子或花椰菜花叶病毒 35S 启动子 (CaMV35S)。实时 PCR 的结果表明，在 16 小时光照/8 小时黑暗光周期下，MSH2-RNAi 番茄转基因植物的叶片中基因转录物含量相对于未转化对照为 0%至 3%。然而，在这种光照模式下，MSH2-RNAi 转基因植物生长缓慢，开花不良，并且不能形成种子。在 12 小时光照/12 小时黑暗光周期下培养时，MSH2-RNAi 转基因植物的基因转录物含量相对于未转化对照为 3%至 42%。在这些条件下，大多数带有 CaMV35S 启动子驱动的 RNAi 构建体的 MSH2-RNAi 转基因植物和一个带有 pro-SmAMP2 启动子驱动的 RNAi 构建体的 MSH2-RNAi 转基因植物形成了 F1 杂种种子。在 12 小时光照/12 小时黑暗光周期条件下，大多数 F1 转基因杂种的基因转录物含量在 3%至 34%之间，并形成了 F1 后代，这使得评估减数分裂重组频率成为可能。我们表明，与对照相比，MSH2-RNAi 番茄转基因植物中基因的有效抑制并不与减数分裂重组的增加相关，而是刺激植物的不育。我们确定，与 16 小时光照/8 小时黑暗光周期相比，番茄植物中基因的表达在 12 小时光照/12 小时黑暗光周期下约高 50 倍。我们讨论了，在对开花日长不敏感的番茄植物中，改变光照时间可能是在一定范围内控制减数分裂重组频率的一种手段。
