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In wine grape, the multi-functional cytochrome P450 enzyme VvCYP76F14 sequentially catalyzes the formation of linalool-derived compounds, including ( E )-8-hydroxylinalool, ( E )-8-oxolinalool, and ( E )-8-carboxylinalool, which are crucial precursors for the wine bouquet. However, molecular basis towards VvCYP76F14 in regulating the wine bouquet precursor production remain unknown. In this study, both wine bouquet precursor contents and catalytic activities of VvCYP76F14s varied among the three different wine bouquet type varieties. Subcellular localization analysis revealed that VvCYP76F14s are predominantly localized in the endoplasmic reticulum. Notably, a maltose-binding protein (MBP) fusion-tag was added to each of the three VvCYP76F14 proteins in the Escherichia coli expression system, significantly induced the concentration of the MBP-VvCYP76F14 fusion proteins. Site-directed mutation of 4 amino acid residues (I120L, L298V, E378G, and T389A) in VvCYP76F14 resulted in a significant decrease in VvCYP76F14 enzymatic activities, respectively. Furthermore, the transient expression of VvCYP76F14 cloned from ‘Yanniang No.2’ significantly increased the levels of ( E )-8-hydroxylinalool, 8-oxolinalool, and ( E )-8-carboxylinalool compounds in the transformed ‘Yanniang No.2’, ‘Italian Riesling’, and ‘Marselan’ berries, respectively. In conclusion, VvCYP76F14 dominates the production of wine bouquet precursors and could be a fingerprint marker for screening superior hybrid offspring with desired levels of wine bouquet precursors.

Salinity (NaCl) stress impairs plant growth and inflicts severe crop losses. In roots, increasing extracellular NaCl causes Ca2+ influx to elevate cytosolic free Ca2+ ([Ca2+]cyt) as a second messenger for adaptive signaling. Amplification of the signal involves plasma membrane reduced nicotinamide adenine dinucleotide phosphate oxidase activation, with the resultant reactive oxygen species triggering Ca2+ influx. The genetic identities of the Ca2+-permeable channels involved in generating the [Ca2+]cyt signal are unknown. Potential candidates in the model plant Arabidopsis (Arabidopsis thaliana) include annexin1 (AtANN1). Here, luminescent detection of [Ca2+]cyt showed that AtANN1 responds to high extracellular NaCl by mediating reactive oxygen species-activated Ca2+ influx across the plasma membrane of root epidermal protoplasts. Electrophysiological analysis revealed that root epidermal plasma membrane Ca2+ influx currents activated by NaCl are absent from the Atann1 loss-of-function mutant. Both adaptive signaling and salt-responsive production of secondary roots are impaired in the loss-of-function mutant, thus identifying AtANN1 as a key component of root cell adaptation to salinity.

In plants, ferritin proteins play an important role in iron (Fe) storage which contributes to plant growth and development. However, the biological functions of ferritins in fruit trees are essentially unknown. In this study, three Ferritin genes were isolated from ‘Zhentong No. 3’ peach, which were named PpFer1-PpFer3. The expression levels of these genes were different in distinct tissues/organs. Notably, PpFer1 was the most abundantly expressed Ferritin family gene in all tested tissues of ‘Zhentong No. 3’ peach; its expression levels were significantly enhanced throughout the entire peach seedling under Fe toxicity and H2O2 stress, particularly in the leaves. In addition, over-expression of PpFer1 was effective in rescuing the retarded growth of Arabidopsis fer1-2 knockout mutant, embodied in enhanced fresh weight, primary root length, lateral root numbers, total root length, total leaf chlorophyll, stomatal conductance (Gs), net photosynthetic rate (Pn), transpiration rate, and tissue Fe concentration. This study provides insights into understanding the molecular mechanisms of Fe storage and sequestration in perennial fruit trees.

Damage can be signalled by extracellular ATP (eATP) using plasma membrane (PM) receptors to effect cytosolic free Ca 2+ ([Ca 2+ ] cyt) increase as a second messenger. The downstream PM Ca 2+ channels remain enigmatic. Here, the Arabidopsis thaliana Ca 2+ channel subunit Cyclic Nucleotide-Gated Channel2 (CNGC2) was identified as a critical component linking eATP receptors to downstream [Ca 2+ ] cyt signalling in roots.  eATP-induced changes in single epidermal cell PM voltage and conductance were measured electrophysiologically, changes in root [Ca 2+ ] cyt were measured with aequorin and root transcriptional changes were determined by qRT-PCR. Two cngc2 loss of function mutants were used: cngc2-3 and dnd1 (which expresses cytosolic aequorin).  eATP-induced transient depolarisation of Arabidopsis root elongation zone epidermal PM voltage was Ca 2+-dependent, requiring CNGC2 but not CNGC4 (its channel co-subunit in immunity signalling). Activation of PM Ca 2+ influx currents also required CNGC2. The eATP-induced [Ca 2+ ] cyt increase and transcriptional response in cngc2 roots were significantly impaired.  CNGC2 is required for eATP-induced epidermal Ca 2+ influx, causing depolarisation leading to [Ca 2+ ] cyt increase and damage-related transcriptional response.

In plants, ferritin proteins play an important role in iron (Fe) storage which contributes to plant growth and development. However, the biological functions of ferritins in fruit trees are essentially unknown. In this study, three genes were isolated from 'Zhentong No. 3' peach, which were named . The expression levels of these genes were different in distinct tissues/organs. Notably, was the most abundantly expressed Ferritin family gene in all tested tissues of 'Zhentong No. 3' peach; its expression levels were significantly enhanced throughout the entire peach seedling under Fe toxicity and H O stress, particularly in the leaves. In addition, over-expression of was effective in rescuing the retarded growth of knockout mutant, embodied in enhanced fresh weight, primary root length, lateral root numbers, total root length, total leaf chlorophyll, stomatal conductance ( ), net photosynthetic rate ( ), transpiration rate, and tissue Fe concentration. This study provides insights into understanding the molecular mechanisms of Fe storage and sequestration in perennial fruit trees.

In plants, ferritin proteins play an important role in iron (Fe) storage which contributes to plant growth and development. However, the biological functions of ferritins in fruit trees are essentially unknown. In this study, three Ferritin genes were isolated from ‘Zhentong No. 3’ peach, which were named PpFer1-PpFer3. The expression levels of these genes were different in distinct tissues/organs. Notably, PpFer1 was the most abundantly expressed Ferritin family gene in all tested tissues of ‘Zhentong No. 3’ peach; its expression levels were significantly enhanced throughout the entire peach seedling under Fe toxicity and H[sub.2] O[sub.2] stress, particularly in the leaves. In addition, over-expression of PpFer1 was effective in rescuing the retarded growth of Arabidopsis fer1-2 knockout mutant, embodied in enhanced fresh weight, primary root length, lateral root numbers, total root length, total leaf chlorophyll, stomatal conductance (G[sub.s] ), net photosynthetic rate (P[sub.n] ), transpiration rate, and tissue Fe concentration. This study provides insights into understanding the molecular mechanisms of Fe storage and sequestration in perennial fruit trees.

Salinity (NaCl) stress impairs plant growth and inflicts severe crop losses. In roots, increasing extracellular NaCl causes Ca 2+ influx to elevate cytosolic free Ca 2+ ([Ca 2+ ] cyt ) as a second messenger for adaptive signaling. Amplification of the signal involves plasma membrane reduced nicotinamide adenine dinucleotide phosphate oxidase activation, with the resultant reactive oxygen species triggering Ca 2+ influx. The genetic identities of the Ca 2+ -permeable channels involved in generating the [Ca 2+ ] cyt signal are unknown. Potential candidates in the model plant Arabidopsis (Arabidopsis thaliana) include annexin1 (AtANN1). Here, luminescent detection of [Ca 2+ ] cyt showed that AtANN1 responds to high extracellular NaCl by mediating reactive oxygen species-activated Ca 2+ influx across the plasma membrane of root epidermal protoplasts. Electrophysiological analysis revealed that root epidermal plasma membrane Ca 2+ influx currents activated by NaCl are absent from the Atann1 loss-of-function mutant. Both adaptive signaling and salt-responsive production of secondary roots are impaired in the loss-of-function mutant, thus identifying AtANN1 as a key component of root cell adaptation to salinity.

NADPH oxidase activity is involved in plant adaptation and development. The reactive oxygen species sourced by NADPH oxidase activity may contribute to wall strength and protoplast volume adjustment. Root hair bulge apices of the NADPH oxidase mutant rhd2/Atrbohc were more robust than the kjk cellulose synthase mutant, but burst more readily than the wild type (WT). Root epidermal wall appeared impaired in rhd2/Atrbohc, as revealed by the number of protoplasts released by wall-degrading enzymes. Root hair bulges of rhd2/Atrbohc burst more than the WT when challenged in situ with hypo-osmotic low ionic strength medium. Inhibition of NADPH oxidase activity with diphenylene iodonium caused WT to phenocopy the rhd2/Atrbohc bursting in response to hypo-osmotic shock. This implicates RHD2/AtRBOHC in softening the cell wall to permit protoplast expansion. Overall, the results point to a role for RHD2/AtRBOHC in contributing to wall strength.