Changes in Ascorbate Content and Its Metabolism Gene Expression Pattern in Garlic Allium sativum L. Leaves in Response to Cold Stress

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Abstract

In two winter garlic (Allium sativum L.) cultivars, the expression of genes encoding key enzymes of the L-galactose pathway of ascorbate (AA) biosynthesis (VTC2, GPP, GalDH, GalLDH) and genes encoding AA recycling from oxidized forms (MDHAR1, MDHAR4, MDHAR5) was determined in the dynamics (0, 2, 4, 6, and 24 h) of short-term (24 h) cold (2°C) stress. It was found that in the leaf tissue of both cultivars (Sarmat and Dubkovsky), the level of VTC2 transcripts steadily increased during stress. The cultivars are also similar in the expression response to stress of the GalLDH and MDHAR1 genes, but differ in the reaction of the GPP, GalDH, MDHAR4 and MDHAR5. In the leaf tissue of both cultivars, the content of AA, carotenoids (total) and chlorophylls (a and b) was determined in the dynamics of cold stress. It was shown that the amount of all analyzed metabolites is initially higher in the cv. Sarmat than in the cv. Dubkovsky, and changes in the stress dynamics in a cultivar-specific manner. A positive correlation was shown between the amount of AA and the level of expression of the GalDH gene for both cultivars.

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About the authors

T. M. Seredin

Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences; Federal Scientific Vegetable Center

Email: michel7753@mail.ru
Russian Federation, Moscow, 119071; Moscow oblast, VNIISSOK village, 143080

A. V. Shchennikova

Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences

Email: michel7753@mail.ru
Russian Federation, Moscow, 119071

E. Z. Kochieva

Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences

Email: michel7753@mail.ru
Russian Federation, Moscow, 119071

M. A. Filyushin

Federal Research Centre “Fundamentals of Biotechnology” of the Russian Academy of Sciences

Author for correspondence.
Email: michel7753@mail.ru
Russian Federation, Moscow, 119071

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2. Fig. 1. Changes in the content of AC (mg/g of crude weight (FW)), chlorophylls a and b, vitamin C, and the amount of carotenoids in garlic roots of Sarmat and Dubkovsky varieties 2, 4, 6, and 24 hours (h) after the start of cold treatment (2 ° C); *p < 0.05 is a significant difference from the initial, pre–stress (0 h) value.

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3. 2. Changes in the expression levels of ascorbate biosynthesis (AsVTC2, AsGPP, AsGalDH, AsGalLDH) and recycling (AsMDHAR1, AsMDHAR4, AsMDHAR5) genes in garlic roots of Sarmat and Dubkovsky varieties 2, 4, 6, and 24 h (h) after the start of cold treatment. The data are normalized to the initial, pre-stress (0 h) value of the expression level, taken as 1 (the value of gene expression at points 2, 4, 6 and 24 h is recalculated proportionally). The color range from blue to red corresponds to the dynamics of gene expression from decrease to increase (vs 1 h).

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4. Fig. 3. Estimation of the correlation between the content of AK (mg/g of crude weight (FW)) and the relative level of transcripts of A K-pathway genes; significant correlation at p < 0.05. On the abscissa axis is the vitamin C content, mg/g FW, on the ordinate axis is the relative expression of the gene.

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