bioRxiv Science⌕ Search

Biology subjects

Eshel, D.

Publications and source records attributed to Eshel, D..

6 recordsLinked to original sources

Dual Knockout of StAMY23 and StVINV Improves Postharvest Storage Traits in Potato

Starch is the primary carbohydrate reserve in potato (Solanum tuberosum L.) tubers and a critical determinant of their industrial value. The rate of starch degradation during postharvest storage directly influences key traits such as endodormancy (ED) duration and cold-induced sweetening (CIS), which affect sprouting behavior. In this study, we used CRISPR/Cas9 genome editing to knockout StAMY23, a gene encoding -amylase involved in starch breakdown. stamy23 plants exhibited higher yield and extended tuber ED postharvest, without significantly altering CIS or starch granule content. To further reduce CIS, we knockout StAMY23 in VACUOLAR INVERTASE knockout (stvinv) backgrounds, generating stamy23/stvinv double-knockouts plants. These lines showed significantly reduced CIS, prolonged ED, and elevated starch content, along with altered starch granule content. Collectively, our findings demonstrate that simultaneous downregulation of StAMY23 and StVINV can additively enhance desirable postharvest traits, providing a promising strategy for improving potato storage quality through precision genome editing.

plant biology↗

Vacuolar invertase knockout enhances drought tolerance in potato plants

Drought stress is one of the most critical abiotic constraints limiting crop productivity worldwide, exacerbated by ongoing climate change and increasingly frequent extreme weather events. Stomatal regulation and osmoprotective sugar accumulation are critical adaptive mechanisms for plant survival under drought stress. Here, we characterize the enhanced drought resilience observed in CRISPR/Cas9-mediated potato, mutants in their vacuolar invertase gene (StVInv). Knockout plants exhibited improved performance under progressive drought stress and during rewatering drought, maintaining higher stomatal conductance, elevated transpiration rates, and superior photosynthetic efficiency compared to wild-type (WT) plants. These improved performance under similar transpiration rate led to higher agronomic water-use efficiency (AWUE) in stvinv plants resulting in greater biomass production despite reduced water availability. Metabolomic profiling revealed distinct adaptive strategies; stvinv plants preferentially accumulated galactinol and raffinose, indicating enhanced raffinose family oligosaccharide (RFO) metabolism. Furthermore, stvinv plants displayed lower levels of abscisic acid (ABA) and its catabolites under drought, suggesting a moderated ABA response facilitating a more risk-taking growth strategy that supports sustained growth and physiological stability. Our findings identify targeted metabolic and hormonal adjustments underlying drought resilience in potato plant, offering promising strategies for enhancing crop performance under water-limited conditions.

plant biology↗

Sugar units, not chilling units, control endodormancy duration in plants; potato tuber as a case study

Endodormancy (ED) is a crucial stage in the life cycle of many perennial plants, regulated by genetic and environmental factors. Chilling units, growth regulators, and nutrient supply are considered inducers of ED release, but the mechanism governing ED duration is poorly understood. The potato tuber has been used as a model system to study metabolic processes associated with ED release. Cold-induced sweetening is a well-known response of the tuber to chilling. Here, we found that cold stress induces an increase in sugar units in association with plasmodesmatal closure in the dormant bud cells. Tuber sweetening was associated with shortened ED duration after cooling. Heat exposure also caused sugar unit accumulation followed by faster ED release. A logistic function was developed to predict ED duration based on sugar unit measurements. We discovered that ED release is better correlated with the accumulation of sugar units compared to chilling units. CRISPR/Cas9 knockout of the vacuolar invertase gene (StVInv) induced longer ED, but only in cultivars in which the mutation modified the level sugar units. Our results suggest that sugar units are better predictors of vegetative bud ED duration than chilling units.

plant biology↗

Sucrose promotes etiolated stem branching through activation of cytokinin accumulation followed by vacuolar invertase activity

The potato (Solanum tuberosum L.) tuber is a swollen stem. Sprouts growing from the tuber nodes represent dormancy release and loss of apical dominance. We recently identified sucrose as a key player in triggering potato stem branching. To decipher the mechanisms by which sucrose induces stem branching, we investigated the nature of the inducing molecule and the involvement of vacuolar invertase (VInv) and the plant hormone cytokinin (CK) in this process. Sucrose was more efficient at enhancing lateral bud burst and elongation than either of its hexose moieties (glucose and fructose), or a slowly metabolizable analog of sucrose (palatinose). Sucrose feeding induced expression of the sucrose transporter gene SUT2, followed by enhanced expression and activity of VInv in the lateral bud prior to its burst. We observed a reduction in the number of branches on stems of VInv-RNA interference lines during sucrose feeding, suggesting that sucrose breakdown is needed for lateral bud burst. Sucrose feeding led to increased CK content in the lateral bud base prior to bud burst. Inhibition of CK synthesis or perception inhibited the sucrose-induced bud burst, suggesting that sucrose induces stem branching through CK. Together, our results indicate that sucrose is transported to the bud, where it promotes bud burst by inducing CK accumulation and VInv activity.

plant biology↗

Endogenous sugar level is associated with differential heat tolerance in onion bulb scales

Postharvest heat treatment stimulates desiccation and browning of outer scales of onion (Allium cepa. L) bulb to dry papery skins. Inner scales resist the heat treatment, as evidenced by high moisture levels. During heating, inner scales showed increasing soluble sugar levels followed by higher osmolarity, vs. a dramatic decrease in both in the outer scales. Exogenous feeding of outer scales with sucrose, glucose or fructose solutions before heat treatment reduced water loss during heating, suggesting a role for soluble sugars in water retention and therefore, heat tolerance. Vacuolar invertase (VInv) is a key enzyme regulating the levels of sucrose, glucose and fructose in plant tissue. VInv-silencing in potato plants prevented the accumulation of reducing sugars in heated leaves, increasing water loss. In onion outer scales, VInv activity increased during heating but reducing sugars decreased, possibly due to their rapid metabolism during scale senescence to form skin. Transcriptomic analysis demonstrated upregulation of genes involved in lignin biosynthesis and secondary cell-wall formation in outer scales during heat exposure, and upregulation of genes involved in energy-related pathways in inner scales. This study reveals the dual role of soluble sugars in different onion scales, as osmoprotectants or building blocks, under heat stress. HighlightHexose formation in the inner scales of onion is associated with heat tolerance, while in the outer scales, the hexoses are metabolized for onion skin formation.

plant biology↗

Vacuolar processing enzyme translocates to the vacuole through the autophagy pathway to induce programmed cell death

The caspase-like vacuolar processing enzyme (VPE) is a key factor in programmed cell death (PCD) associated with plant stress responses. Growth medium lacking a carbon source and dark conditions caused punctate labeling of 35S::VPE1-GFP (StVPE1-GFP) in potato leaves. Carbon starvation of BY-2 cells induced higher VPE activity and PCD symptoms. Growing VPE-RNAi BY-2 cells without sucrose reduced VPE activity and prevented PCD symptoms. During extended exposure to carbon starvation, VPE expression and activity levels peaked, with a gradual increase in BY-2 cell death. Histological analysis of StVPE1-GFP in BY-2 cells showed that carbon starvation induces its translocation from the endoplasmic reticulum to the central vacuole, through tonoplast engulfment. Exposure of BY-2 culture to the autophagy inhibitor concanamycin A caused autophagic bodies accumulation in the cell vacuole. Such accumulation did not occur in the presence of 3-methyladenine, an inhibitor of early-stage autophagy. BY-2 cells constitutively expressing StATG8IL-RFP, an autophagosome marker, showed colocalization with the StVPE1-GFP protein in the cytoplasm and vacuole. RNAi silencing of the core autophagy component ATG4 in BY-2 cells reduced VPE activity and cell death. These results are the first to suggest that VPE translocates to the cell vacuole through the autophagy pathway, leading to PCD.\n\nOne sentence summaryCarbon starvation induced programmed cell death by trafficking vacuolar processing enzyme through the autophagy pathway to the vacuole.

plant biology↗