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An early light-inducible protein (GbELIP) isolated from immature ginkgo leaves by rapid amplification of cDNA ends method contains a 570 bp open reading frame encoding a 189 amino acid protein, with significant homology to early light-inducible proteins from other plant species. The genomic DNA of GbELIP gene contains three exons and two introns. Southern blotting revealed that GbELIP is a singlecopy gene in ginkgo. Light, defense and stress responsive element were found in the promoter region of GbELIP. GbELIP expressional patterns were detected by Real-time PCR at various conditions. ELIP expressions were variable at different time points of 1 day and increasing significantly at the commencement of light illumination then dropping to a basal level gradually. ELIP expression in small size leaves (1–1.5 cm width and 7-day age) was higher then middle (2–3 cm width and 15-day age) and large (4 cm width and 30-day age) size leaves. It revealed that ELIP expression was under the control of developmental regulation. Under temperature and light intensity treatments, GbELIP transcripts accumulation may relate to photoinhibition. The peak expression of GbELIP appeared later by chilling than heating Meanwhile, GbELIP expression under high light was higher than low light at both two temperatures. GbELIP expression was also increased by wounding and methyl jasmonate (Me-JA) treatment, but not corresponded with increasing malondialdehyde (MDA) concentration. In conclusion, GbELIP appears to be induced by an imbalance between chlorophyll formation and degradation during development or under abiotic stress. These results suggest that GbELIP may function in response to environmental signals, possibly regulating responses to abiotic stresses.
Ginkgo suspension cells were used to investigate the mechanism that governs the shift between primary and secondary metabolism under NaCl elicitation. The production of three flavonol glycosides, chlorophyll fluorescence, ion content, the antioxidant system, and the cellular ultrastructure in the presence of NaCl doses from 5 to 175 mM were examined. At low salt doses (5–50 mM), cell growth and flavonol glycosides accumulation were stimulated without damaging cell structure or inducing oxidative stress by maintaining high K⁺ and chlorophyll content. At moderate salt doses (75–125 mM), the cells could withstand the salt stress without an impact on survival by changing internal cellular structure, maintaining high levels of K⁺ and Ca²⁺ and increasing anti-oxidative enzyme activities rather than flavonol glycosides to counteract the inhibition of the photosystem II, the accumulation of Na⁺ and hydrogen peroxide (H₂O₂) in the cells. This allowed cells to divert their metabolism from growth to defense-related pathways and tolerate NaCl stress. At higher salinity (150–175 mM), the cellular structure was damaged, and the high Na⁺ and low K⁺ content led to osmotic stress, and therefore, the stimulation of peroxidase (POD) and catalase (CAT) was not enough to cope with high H₂O₂ accumulation. The high production of flavonol glycosides may be a response of elicitation stimulation to serious damage at 175 mM NaCl. In conclusion, the use of 175 mM NaCl may be desirable for the induction of flavonol glycoside production in Ginkgo suspension cells.
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