Ectopic expression of a novel cold-resistance protein 1 from Brassica oleracea promotes tolerance to chilling stress in transgenic tomato

Abstract

Cold stress is considered as one of the major environmental factors that adversely affects the plant growth and distribution. Therefore, there arises an immediate need to cultivate effective strategies aimed at developing stress-tolerant crops that would boost the production and minimise the risks associated with cold stress. In this study, a novel cold-responsive protein1 (BoCRP1) isolated from Brassica oleracea was ectopically expressed in a cold susceptible tomato genotype Shalimar 1 and its function was investigated in response to chilling stress. BoCRP1 was constitutively expressed in all the tissues of B. oleracea including leaf, root and stem. However, its expression was found to be significantly increased in response to cold stress. Moreover, transgenic tomato plants expressing BoCRP1 exhibited increased tolerance to chilling stress (4 °C) with an overall improved rate of seed germination, increased root length, reduced membrane damage and increased accumulation of osmoprotectants. Furthermore, we observed increased transcript levels of stress responsive genes and enhanced accumulation of reactive oxygen species scavenging enzymes in transgenic plants on exposure to chilling stress. Taken together, these results strongly suggest that BoCRP1 is a promising candidate gene to improve the cold stress tolerance in tomato.

ICE-CBF-COR Signaling Cascade and Its Regulation in Plants Responding to Cold Stress

“…Recently, Kashyap et al [73] showed an ICE homolog, BjICE53, to be involved in the cold signaling pathway in Brassica juncea. They revealed conserved domains and motifs that bind to the CRT/DRE motifs of BjCBF for the expression of downstream cold-regulatory genes [74][75][76]. Another study in Chrysanthemum morifolium, "Jinba", demonstrated that overexpression of CmICE2 in transgenic Arabidopsis induces the expression of downstream cold regulatory genes (AtCBF1/2, AtCOR6.6a/414, and AtKIN1), leading to cold stress tolerance through increased proline contents, superoxide dismutase (SOD) activities, and elevating catalase (CAT) levels [77].…”

Fine-Tuning Cold Stress Response Through Regulated Cellular Abundance and Mechanistic Actions of Transcription Factors

“…In response to cold stress, the abscisic acid (ABA), jasmonic acid (JA), and indole acetic acid (IAA) are among the phytohormones involved in activating CBF-dependent signaling pathway ( Wang et al, 2021 ; Zhou et al, 2021 ; Fu et al, 2022 ). The interaction between CBF and CRT/DRE motif regulates the downstream expression of an array of COR genes, which belong to KIN (cold induced), RD (responsive to desiccation), LTI (low temperature induced), and ERD (early dehydration inducible; Wani et al, 2021 ). The regulated products of COR genes include osmo-protectants biosynthetic enzymes, late embryogenesis abundant (LEA) proteins, TFs, protein kinases, proteins associated with lipid metabolism, proteins for hormone responses, cell wall modifiers, and chloroplast proteins ( Liu et al, 2019 ).…”

Isolation of high-quality RNA for high throughput applications from secondary metabolite-rich Crocus sativus L.

“…For each qPCR reaction, 1 μl of cDNA diluted by a factor of 10 was mixed in a 0.2 ml RNase free tube containing 6 μl of 2X Syber green, 0.3 μl each of forward and universal reverse primer, and 4.4 ml Milli-Q water. The reaction mixture was run in lightCycler480-II (Roach) and the fold chnage was calculated using 2 −∆∆CT method [ 16 ].…”