A Vacuolar Invertase CsVI2 Regulates Sucrose Metabolism and Increases Drought Tolerance in Cucumis sativus L.

Vacuolar invertase (VI) can irreversibly degrade sucrose into glucose and fructose and involve in plants abiotic-stress-tolerance. Cucumber (Cucumis sativus L.) is susceptible to drought stress, especially during the seedling stage. To date, the involvement of VI in drought tolerance in cucumber seedlings is in urgent need of exploration. In the present study, a cucumber vacuolar invertase gene, CsVI2, was isolated and functionally characterized. The results showed that (1) CsVI2 showed vacuolar invertase activity both in vivo and in vitro; (2) the transcript level of CsVI2, along with VI activity, was significantly induced by drought stress. Moreover, the expression of sucrose synthase 3 (CsSUS3) was increased and that of sucrose phosphate synthase 1 (CsSPS1) was decreased after exposure to drought stress, which was followed by an increase in sucrose synthase activity and a decrease in sucrose phosphate synthase activity; (3) CsVI2-overexpressing transformed cucumber seedlings showed enhanced vacuolar invertase activity and drought tolerance and 4) protein–protein interaction modelling indicated that a cucumber invertase inhibitor, CsINVINH3, can interact with CsVI2. In summary, the results indicate that CsVI2 as an invertase can regulate sucrose metabolism and enhance drought stress in cucumber seedlings.

Functional Characterization of a Cucumber (Cucumis sativus L.) Vacuolar Invertase, CsVI1, Involved in Hexose Accumulation and Response to Low Temperature Stress

Cucumber (Cucumis sativus L.), an important vegetable plant species, is susceptible to low temperature stress especially during the seedling stage. Vacuolar invertase (VI) plays important roles in plant responses to abiotic stress. However, the molecular and biochemical mechanisms of VI function in cucumber, have not yet been completely understood and VI responses to low temperature stress and it functions in cold tolerance in cucumber seedlings are also in need of exploration. The present study found that hexose accumulation in the roots of cucumber seedlings under low temperature stress is closely related to the observed enhancement of invertase activity. Our genome-wide search for the vacuolar invertase (VI) genes in cucumber identified the candidate VI-encoding gene CsVI1. Expression profiling of CsVI1 showed that it was mainly expressed in the young roots of cucumber seedlings. In addition, transcriptional analysis indicated that CsVI1 expression could respond to low temperature stress. Recombinant CsVI1 proteins purified from Pichia pastoris and Nicotiana benthamiana leaves could hydrolyze sucrose into hexoses. Further, overexpression of CsVI1 in cucumber plants could increase their hexose contents and improve their low temperature tolerance. Lastly, a putative cucumber invertase inhibitor was found could form a complex with CsVI1. In summary, these results confirmed that CsVI1 functions as an acid invertase involved in hexose accumulation and responds to low temperature stress in cucumber seedlings.

Advances about the Roles of Membranes in Cotton Fiber Development

Cotton fiber is an extremely elongated single cell derived from the ovule epidermis and is an ideal model for studying cell development. The plasma membrane is tremendously expanded and accompanied by the coordination of various physiological and biochemical activities on the membrane, one of the three major systems of a eukaryotic cell. This review compiles the recent progress and advances for the roles of the membrane in cotton fiber development: the functions of membrane lipids, especially the fatty acids, sphingolipids, and phytosterols; membrane channels, including aquaporins, the ATP-binding cassette (ABC) transporters, vacuolar invertase, and plasmodesmata; and the regulation mechanism of membrane proteins, such as membrane binding enzymes, annexins, and receptor-like kinases.

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 of sugar units. Our results suggest that sugar units are better predictors of vegetative bud ED duration than chilling units.

Performance of Hevea brasiliensis under drought conditions on osmoregulation and antioxidant activity through evaluation of vacuolar invertase and reducing sugars

Rubber tree cultivation is limited in many regions by abiotic factors such as drought. We investigated the biochemical mechanisms responsible for responses to, and recovery from, drought conditions during the establishment phase of four high latex producing rubber tree clones (RRIM600, IAC40, PR255 and GT1). Five-month-old plants were exposed to 32 days of water restriction, followed by 15 days of soil rehydration. Leaf area, as well as their osmolyte accumulations, saccharolytic enzyme activity, and oxidative stress markers, were accompanied. Although clones IAC40 and PR255 responded more precociously to drought conditions, halting leaf expansion before clones GT1 and RRIM600, they demonstrated slow recuperation after reestablishing irrigation. The greater tolerances of clones RRIM600 and GT1 to drought conditions were related to greater vacuolar invertase (VINV) activity in their leaves, which guaranteed more significant accumulations of vacuolar reducing sugars (RS). Similar to RS, glycine betaine accumulations were related to osmoprotection and to reducing oxidative damage (lipidic peroxidation) caused by water deficit conditions. The observed decreases in cytosol neutral invertase (AINV) and cell wall insoluble invertase (CWINV) activities, which resulted in cytosol hexose decreases, may be related to increases in antioxidant enzyme (superoxide dismutase and ascorbate peroxidase) activities in the leaves in response to water deficit conditions. As such, the introduction of specific sugars (RS) and the modulation of key carbon metabolism enzymes, such as VINV, are promising strategies for promoting drought tolerance in rubber tree clones.

StRAP2.3, an ERF‐VII transcription factor, directly activates StInvInh2 to enhance cold-induced sweetening resistance in potato

Potato invertase inhibitor (StInvInh2) positively regulates cold-induced sweetening (CIS) resistance by inhibiting the activity of vacuolar invertase. The distinct expression patterns of StInvInh2 have been thoroughly characterized in different potato genotypes, but the related CIS ability has not been characterized. The understanding of the regulatory mechanisms that control StInvInh2 transcription is unclear. In this study, we identified an ERF‐VII transcription factor, StRAP2.3, that directly regulates StInvInh2 to positively modulate CIS resistance. Acting as a nuclear-localized transcriptional activator, StRAP2.3 directly binds the ACCGAC cis-element in the promoter region of StInvInh2, enabling promoter activity. Overexpression of StRAP2.3 in CIS-sensitive potato tubers induced StInvInh2 mRNA abundance and increased CIS resistance. In contrast, silencing StRAP2.3 in CIS-resistant potato tubers repressed the expression of StInvInh2 and decreased CIS resistance. We conclude that cold-responsive StInvInh2 is due to the binding of StRAP2.3 to the ACCGAC cis-element in the promoter region of StInvInh2. Overall, these findings indicate that StRAP2.3 directly regulates StInvInh2 to positively modulate CIS resistance, which may provide a strategy to improve the processing quality of potatoes.

Regulation of Maize Kernel Carbohydrate Metabolism by Abscisic Acid Applied at the Grain-Filling Stage at Low Soil Water Potential

A water deficit during the grain-filling stage increases the frequency of yield loss in maize (Zea mays L.). Abscisic acid (ABA) plays a regulatory role in many stages of plant growth; however, its effects on sucrose-metabolizing enzyme activities under stress are poorly understood. The activities of cell-wall-bound acid invertase, vacuolar invertase, cytoplasmic invertase, and sucrose synthase decreased continuously under drought stress, whereas ABA treatment partially restored these activities. In addition, the increase and development of sucrose content under drought stress were related to invertase activity. Up-regulation of the activities and gene expression of cell-wall-bound acid invertase and vacuolar invertase with ABA treatment contributed to the increase in the number of rows and number of grains per row. Furthermore, ABA inhibited the increase in the length of the bald tip. Compared with the control group, water stress significantly reduced the yield index, with the lowest yield index on the 10th day of stress. These results suggest that the increase in ABA-induced sucrose-metabolizing enzyme activity might be an effective mechanism to improve maize drought resistance at the grain-filling stage.

PpINH1, an invertase inhibitor, interacts with vacuolar invertase PpVIN2 in regulating the chilling tolerance of peach fruit

Sucrose metabolism, particularly the decomposition of sucrose by invertase, plays a central role in plant responses to cold stress. Invertase inhibitors (INHs) evolved in higher plants as essential regulators of sucrose metabolism. By limiting invertase activity, INHs keep cellular sugar levels elevated, which provides enhanced protection to plants under stress. Our results showed that the expression of PpVIN2, the only vacuolar invertase (VIN) gene in peach fruit sensitive to chilling temperatures, increases significantly during cold storage, while VIN enzyme activity increases more modestly. We also found that peach fruit transiently overexpressing PpINH1 had decreased VIN activity. Interactions of PpINH1 and PpVIN2 with recombinant proteins were shown by yeast two-hybrid assays and bimolecular fluorescence complementation assays, as well as in vitro. During cold storage, trehalose-treated peach fruit had significantly increased PpINH1 expression, decreased VIN enzyme activity, and significantly higher sucrose content than did untreated fruit. As a result, the treated fruit had enhanced resistance to chilling injury. Collectively, our data show that the post-translational repression of VIN enzyme activity by PpINH1 helps maintain sucrose levels in peach fruit during cold storage, thereby improving resistance to chilling injury.