Study of the effect of the method of applying cryoprotectors on the preservation of Mentha longifolia seed material during cryopreservation

A. Dodonova
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Abstract

This article is devoted to optimizing the conditions for cryopreservation of seed material of Mentha longifolia. Mentha longifolia is an essential oil plant that is actively used in the perfumery, pharmaceutical and food industries. The cryopreservation experiments use the following endocellular and exocellular cryoprotectants: polyvinylpyrrolidone (PVP), glycerol, and dimethysulfoxide (DMSO). In addition, we study the effect of the growth characteristics of seeds on the preservation during cryopreservation of the method of applying a cryoprotectant: at room temperature and in an ice bath. The initial germination of long-leaved mint seeds is 76.3±2 %, and the germination energy is 70.3±1.8 %. When frozen in liquid nitrogen without the use of cryoprotectants, the germination rate was 64.9±0.8 %, and the germination energy was 18.95±0.2 %. It should be noted that the germination energy indicator turned out to be sensitive to the deposition of seed material at extremely low temperatures. An analysis of the results of maintaining the viability of long-leaved mint seeds when they are frozen in cryoprotectants of various compositions in the case of their introduction at room temperature indicates that the tendency for a sharp decrease in germination energy is observed in all variants of the experiment. The best options for cryoprotectants to preserve the growth characteristics of the seed material are DMSO 2.5 % and PVP 3 % applied in an ice bath. The germination of seeds stored in these cryoprotectants is 85.8±3.1 % and 87.6±2.3 %, respectively. Thus, it is found that the best way to apply cryoprotectants during cryopreservation of long-leaved mint seeds is a temperature close to 0 °C; in this case, both germination and seed germination energy are seriously increased.
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低温保鲜剂对薄荷种子材料低温保存效果的研究
本文对长叶薄荷种子材料的低温保存条件进行了优化。薄荷是一种精油植物,在香料、制药和食品工业中被广泛使用。低温保存实验使用以下细胞内和细胞外低温保护剂:聚乙烯吡咯烷酮(PVP),甘油和二甲基亚砜(DMSO)。此外,我们还研究了在室温和冰浴冷冻保存中使用冷冻保护剂的方法对种子生长特性的影响。长叶薄荷种子的初始发芽率为76.3±2%,萌发能为70.3±1.8%。在液氮中冷冻时,不使用冷冻保护剂,发芽率为64.9±0.8%,萌发能为18.95±0.2%。值得注意的是,萌发能指标对极低温下种子物质的沉积非常敏感。对长叶薄荷种子在室温下被冷冻在不同成分的冷冻保护剂中以保持其活力的结果的分析表明,在实验的所有变体中都观察到萌发能量急剧下降的趋势。为了保持种子材料的生长特性,冷冻保护剂的最佳选择是在冰浴中应用2.5%的DMSO和3%的PVP。冷冻保护剂中种子的发芽率分别为85.8±3.1%和87.6±2.3%。因此,在长叶薄荷种子的低温保存过程中,使用低温保护剂的最佳温度为接近0℃;在这种情况下,发芽率和种子发芽能都大大增加。
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