Isolation and characterization of the trehalose-6-phosphate synthase gene from Locusta migratoria manilensis

Trehalose plays an important role in protecting organisms from various stresses. Trehalose-6-phosphate synthase （TPS） is the key enzyme in trehalose synthesis, but in insects only a few TPS genes have been identified and their function has not been well characterized. To better understand the function of TPS in insects, a complete TPS complementary DNA （eDNA） clone was obtained from the fat body of the locust Locusta migratoria manilensis （GenBank accession number： EU131894）. The full-length cDNA is 2 806 bp, including an open reading frame of 2 442 bp, which encodes an 813 amino acids protein with a calculated molecular weight of 91 976 Daltons and an isoelectric point of 6.14. The deduced amino acid sequence is highly similar to other published insect TPS and its C-terminal also has a region homologous to trehalose phosphate phsophatase （TPP）. Semi-quantitative analysis indicated that the TPS transcript was expressed not only in fat body, but also in gut, hemolymph and leg muscle. These data may facilitate studies of TPS function in insects and improve our understanding of trehalose metabolism.

RNA interference of a trehalose-6-phosphate synthase gene reveals its roles in the biosynthesis of chitin and lipids in Heortia vitessoides(Lepidoptera:Crambidae)

Trehalose 6-phosphate synthase(TPS),an enzyme that hydrolyzes two glucose molecules to yield trchalose,plays a pivotal role in various physiological processes.In this study,we cloned the trehalose-6-phosphate synthase gene(HvTPS)and investigated its expression patterns in various tssues and d:velopmental stages in Heortia vitessoides Moore(Lepidoptera:Crambidac).HvTPS was highly expressed in the fat body and after pupation or before molting.We knocked down TPS in H.vitessoides by RNA interference and found that 3.0μg of dsHvTPS resulted in optimal interference at 24 h and 36 h post-injection and caused a sharp decline in the survival rate during the 5th instar larval-pupal stage and obviously abnormal or lethal phenotypes.Additionally.compared to the controls,TPS activity and trehalose contents were significantly lower and the glucose content was significantly higher 24 h or 36 h after injection with 3.0μg of dsHIvTPS.Furthermore,the silencing of HvTPS suppressed the cxpression of six key genecs in the chitin biosynthesis pathway and one key gene related to lipid catabolism.The expression levels of two genes associated with lipid biosynthesis were upregulated.These results strongly suggest that HvTPS is essential for the normal growth and development of H.vitessoides and provide a reference for further studies of the utility of key genes involved in chitin and lipid biosynthesis for controlling insect development.

Trehalose:A sugar molecule involved in temperature stress management in plants

Trehalose(Tre)is a non-reducing disaccharide found in many species,including bacteria,fungi,invertebrates,yeast,and even plants,where it acts as an osmoprotectant,energy source,or protein/membrane protector.Despite relatively small amounts in plants,Tre concentrations increase following exposure to abiotic stressors.Trehalose-6-phosphate,a precursor of Tre,has regulatory functions in sugar metabolism,crop production,and stress tolerance.Among the various abiotic stresses,temperature extremes(heat or cold stress)are anticipated to impact crop production worldwide due to ongoing climate changes.Applying small amounts of Tre can mitigate negative physiological,metabolic,and molecular responses triggered by temperature stress.Trehalose also interacts with other sugars,osmoprotectants,amino acids,and phytohormones to regulate metabolic reprogramming that underpins temperature stress adaptation.Transformed plants expressing Tre-synthesis genes accumulate Tre and show improved stress tolerance.Genome-wide studies of Tre-encoding genes suggest roles in plant growth,development,and stress tolerance.This review discusses the functions of Tre in mitigating temperature stress—highlighting genetic engineering approaches to modify Tre metabolism,crosstalk,and interactions with other molecules—and in-silico approaches for identifying novel Tre-encoding genes in diverse plant species.We consider how this knowledge can be used to develop temperature-resilient crops essential for sustainable agriculture.

Trehalose-6-phosphate phosphatases are involved in trehalose synthesis and metamorphosis in Bactrocera minax

Trehalose is the principal sugar circulating in the hemolymph of insects,and trehalose synthesis is catalyzed by trehalose-6-phosphate synthase(TPS)and trehalose-6-phosphate phosphatase(TPP).Insect TPS is a fused enzyme containing both TPS do-main and TPP domain.Thus,many insects do not possess TPP genes as TPSs have re-placed the function of TPPs.However,TPPs are widely distributed across the dipteran insects,while the roles they play remain largely unknown.In this study,3 TPP genes from notorious dipteran pest Bactrocera minax(BmiTPPB,BmiTPPCl,and BmiTPPC2)were identified and characterized.The different temporal-spatial expression patterns of 3 BmiTPPs implied that they exert different functions in B.minax.Recombinant BmiTPPs were heterologously expressed in yeast cells,and all purified proteins exhibited enzy-matic activities,despite the remarkable disparity in performance between BmiTPPB and BmiTPPCs.RNA interference revealed that all BmiTPPs were successfully downregulated after double-stranded RNA injection,leading to decreased trehalose content and increased glucose content.Also,suppression of BmiTPPs significantly affected expression of down-stream genes and increased the mortality and malformation rate.Collectively,these results indicated that all 3 BmiTPPs in B.minax are involved in trehalose synthesis and metamor-phosis.Thus,these genes could be evaluated as insecticidal targets for managing B.minax,andevenforotherdipteranpests.

How Do Sugars Regulate Plant Growth and Development？ New Insight into the Role of Treha Iose-6-Phosphate

Plant growth and development are tightly controlled in response to environmental conditions that influence the availability of photosynthetic carbon in the form of sucrose. Trehalose-6-phosphate （T6P）, the precursor of trehalose in the biosynthetic pathway, is an important signaling metabolite that is involved in the regulation of plant growth and development in response to carbon availability. In addition to the plant＇s own pathway for trehalose synthesis, formation of T6P or trehalose by pathogens can result in the reprogramming of plant metabolism and development. Developmental processes that are regulated by T6P range from embryo development to leaf senescence. Some of these processes are regulated in interaction with phytohormones, such as auxin. A key interacting factor of T6P signaling in response to the environment is the protein kinase sucrose non-fermenting related kinase-1 （SnRK1）, whose catalytic activity is inhibited by T6R SnRK1 is most likely involved in the adjustment of metabolism and growth in response to starvation. The transcription factor bZlP11 has recently been identified as a new player in the T6P/SnRK1 regulatory pathway. By inhibiting SnRK1, T6P promotes biosynthetic reactions. This regulation has important consequences for crop production, for example, in the developing wheat grain and during the growth of potato tubers.

SlTPP4 participates in ABA-mediated salt tolerance by enhancing root architecture in tomato

Salinity tolerance is an important physiological index for crop breeding.Roots are typically the first plant tissue to withstand salt stress.In this study,we found that the tomato(Solanum lycopersicum)trehalose-6-phosphate phosphatase(SlTPP4)gene is induced by abscisic acid(ABA)and salt,and is mainly expressed in roots.Overexpression of SlTPP4 in tomato enhanced tolerance to salt stress,resulting in better growth performance.Under saline conditions,SlTPP4 overexpression plants demonstrated enhanced sucrose metabolism,as well as increased expression of genes related to salt tolerance.At the same time,expression of genes related to ABA biosynthesis and signal transduction was enhanced or altered,respectively.In-depth exploration demonstrated that SlTPP4 enhances Casparian band development in roots to restrict the intake of Na^(+).Our study thus clarifies the mechanism of SlTPP4-mediated salt tolerance,which will be of great importance for the breeding of salt-tolerant tomato crops.

Overexpression of the ThTPS gene enhanced salt and osmotic stress tolerance in Tamarix hispida

Trehalose is a non-reducing disaccharide with high stability and strong water absorption properties that can improve the resistance of organisms to various abiotic stresses.Trehalose-6-phosphate synthase(TPS)plays important roles in trehalose metabolism and signaling.In this study,the full-length cDNA of ThTPS was cloned from Tamarix hispida Willd.A phylogenetic tree including ThTPS and 11 AtTPS genes from Arabidopsis indicated that the ThTPS protein had a close evolutionary relationship with AtTPS7.However,the function of At TPS7 has not been determined.To analyze the abiotic stress tolerance function of ThTPS,the expression of ThTPS in T.hispida under salt and drought stress and JA,ABA and GA3 hormone stimulation was monitored by qRT-PCR.The results show that ThTPS expression was clearly induced by all five of these treatments at one or more times,and salt stress caused particularly strong induction of Th TPS in the roots of T.hispida.The ThTPS gene was transiently overexpressed in T.hispida.Both physiological indexes and staining results showed that ThTPS gene overexpression increased salt and osmotic stress tolerance in T.hispida.Overall,the Th TPS gene can respond to abiotic stresses such as salt and drought,and its overexpression can significantly improve salt and osmotic tolerance.These findings establish a foundation to better understand the responses of TPS genes to abiotic stress in plants.

Arabidopsis Coordinates the Diurnal Regulation of Carbon Allocation and Growth across a Wide Range of Photoperiods

In short photoperiods, plants accumulate starch more rapidly in the light and degrade it more slowly at night, ensuring that their starch reserves last until dawn. To investigate the accompanying changes in the timing of growth, Arabidopsis was grown in a range of photoperiods and analyzed for rosette biomass, photosynthesis, respiration, ribosome abundance, polysome loading, starch, and over 40 metabolites at dawn and dusk. The data set was used to model growth rates in the daytime and night, and to identify metabolites that correlate with growth. Modeled growth rates and polysome loading were high in the daytime and at night in long photoperiods, but decreased at night in short photoperiods. Ribosome abundance was similar in all photoperiods. It is discussed how the amount of starch accumulated in the light period, the length of the night, and maintenance costs interact to constrain growth at night in short photoperiods, and alter the strategy for optimizing ribosome use. Significant correlations were found in the day- time and the night between growth rates and the levels of the sugar-signal trehalose 6-phosphate and the amino acid biosynthesis intermediate shikimate, identifying these metabolites as hubs in a network that coordinates growth with diurnal changes in the carbon supply.