This study investigated the effects of endogenous components and extrusion modification on breadfruit starch (BS) digestibility via selective removal of proteins, lipids, pectins, combined with in vitro digestion and multiscale structural characterization. In the non-extruded state, removing endogenous components significantly enhanced starch digestibility, with pectin exerting the strongest inhibitory effect. The resistant starch (RS) content of native BS (29.02 %) decreased to 25.05 % (deproteinized), 10.68 % (defatted), and 6.94 % (depectinized). Depectinization most notably disrupted starch granules, altered short-range order, crystallinity, and chain length distribution. After extrusion, BS formed enzyme-resistant V-type starch-lipid complexes, with RS content increasing to 28.40-38.27 %. The extruded starch showed dense block-like aggregates, reduced amylose leaching, and lower molecular weight. Lipids had the greatest impact on extruded samples, as extruded delipidized starch had the lowest V-type crystallinity (4.59 %) and RS (28.40 %), with altered chain length distribution. This study clarifies endogenous components' regulatory roles in BS digestibility under extrusion, supporting high-RS and low-glycemic index food development.

Nervonic acid (NA), a very long-chain monounsaturated fatty acid, with high value in brain disease prevention and improvement, many studies revealed it could be synthesized in crop seeds via genetic engineering. In recent years, vegetative tissues such as leaves have been proposed as a novel platform for producing triacylglycerol (TAG) and achieving significant results, but the potential for producing NA has not been explored. In this study, three genes, CgKCS from (正体) Cardamine graeca, SLC1-1 from Saccharomyces cerevisiae, and DGAT1 from Arabidopsis thaliana, were selected and under the regulation of the green organizations-specific promoter Rubisco to form multi-gene vector and transformed four varieties rapeseed with different fatty acid profile to explore the potential of NA production in leaves. Those genes have been respectively reported to increase NA production, promote Fatty acids (FAs) combining to the sn-2 of TAG, and improve crop oil content of plant seeds. In this study, NA was not detected in all four varieties of transgenic rapeseed leaves at vegetative, flowering, and seed setting three development stages. Analysis of fatty acid components results shown that linolenic acid (C18:3) is the main fatty acid in leaves, but the available substrates for synthesizing NA are monounsaturated fatty acids, such as C18:1, C20:1, and C22:1, so lacking of monounsaturated fatty acids maybe is the main limiting factor for the production NA in transgenic rapeseed leaves. And then, we further analyzed the feasible strategies to promote NA and other very long-chain fatty acids synthesis, assembly and accumulation. Together, our results provide a clear profile of the fatty acid composition of rapeseed leaves and although it failed to synthesize NA, it also provides referable hints for the production of NA from rapeseed leaves via metabolic engineering.

Nervonic acid (NA), a very-long-chain monounsaturated fatty acid, is known for its benefits in treating neurological diseases and promoting brain health. In this study, we utilized two different receptors, Brassica juncea (B. juncea, rich in erucic acid, C22:1) and Brassica napus (B. napus, high in oleic acid, C18:1), to overproduce NA through systematic metabolic engineering. Two multi-gene vector constructs, Napin-3 and Napin-5 (CgKCS::SLC1-1::DGAT1; CgKCS::SLC1-1::BnFAE1::LdLPAAT::DGAT1), are driven by seed-specific napin promoters. In B. juncea, Napin-3 and Napin-5 expression elevated NA levels to 48.7% and 46.3% in seed oil, respectively, compared to 2.5% in wild types. In B. napus, Napin-3 and Napin-5 expression achieved NA levels of 45% and 39.6%, respectively, while NA is absent in wild types. To our knowledge, this represents the highest NA production in plants to date, with stable oil content and yield, enabling cost-effective NA production. In B. juncea, a significant increase in NA is observed alongside a decrease in C18:1, C20:1, and C22:1 levels; in B. napus, the rise in NA is accompanied by a decrease in C18:1, and an increase in C20:1 and C22:1. These patterns reflect the dynamic equilibrium of fatty acids following NA conversion, influenced by the Dynamic Substrate Tugging (DST) Mechanism, in the form of either an EA-tugging mode or C18:1-tugging mode mechanism, depending on the cellular context. NA is an elongation product derived from C18:1, catalyzed by CgKCS with broad substrate specificity, indicating that plants with high levels of C18:1, similarly to those rich in C22:1, serve as excellent candidates for NA production. This "green factory" for NA production provides strong support for its pharmaceutical, nutraceutical, and industrial applications. The exogenous and the endogenous enzymes coordinate function remodeling of the intra-seed fatty acid elongation flux through the DST strategy, thereby systematically enhancing the synthesis and accumulation efficiency of the target fatty acid.

This study investigated the complexes of gallic acid (GA) with cassava starch (CS), breadfruit starch (BFS), and banana starch (BS)—to evaluate their physicochemical properties, digestion behaviors, and antioxidant activities. The complexation rates for CS-GA, BFS-GA, and BS-GA were 41.00%, 37.41%, and 23.41%, respectively. Complexation enhanced the short-range molecular order and increased the particle size of gelatinized starch, with increments of 37.93%, 31.90%, and 19.81%, respectively. It also reduced thermal stability, gelatinization viscosity, gel hardness, and rapidly digestible starch (RDS) content, while delaying starch retrogradation and increasing resistant starch (RS) and slowly digestible starch (SDS) contents. Complexation protected GA from degradation, resulting in a bioaccessibility exceeding 90% for all complexes. Furthermore, antioxidant assays showed that the decline in DPPH radical scavenging during digestion increased with higher RS content. These findings provide a theoretical foundation and novel insights for the development of functional starch-based foods of these tropical starches.

【Objective】Hainan has a tropical maritime monsoon climate with distinct dry and wet seasons, with concentrated rainfall from June to October. The rainfall is heavy and frequent. Therefore, seasonal waterlogging has become one of the major threats to crops in Hainan. Woody crops have a large biomass and slow response to waterlogging, which is often overlooked. As waterlogging time prolongs,the harm of waterlogging stress becomes increasingly prominent. This study aimed to investigate the responses of two Artocarpus species to different degrees of flooding stress, and comprehensively evaluate their waterlogging tolerance, so as to provide a theoretical basis for the study on waterlogging resistance of woody food crops.【Methods】Jackfruit (Artocarpus heterophyllus Lam.) and breadfruit[Artocarpus altilis (Parkinson) Fosberg], two Artocarpus species, were used as experimental materials, and the pot flooding test was carried out by the‘double pot method'. The experiment was designed as five treatments, i.e., normal water management (seedlings were flooded for 0 h) and flooded for 24 h, 48 h, 72 h, and 96 h. The plants treated with flooding were watered thoroughly to ensure that the water level exceeded the soil surface by about 2 cm. Afterwards, water was replenished daily to keep the roots in a saturated flooded state. After the first watering of the control plants, a certain amount of water should be replenished every 3 days to maintain the overall soil moisture content at 60% to 80% of the maximum field capacity. The changes of biomass, photosynthetic characteristics and antioxidant enzyme activities of jackfruit and breadfruit in different flooding times were investigated, and the waterlogging tolerance was evaluated by factor analysis and membership function methods.【Results】Flooding for 24 h significantly inhibited the aboveground dry weight and photosynthetic parameters of jackfruit seedlings, and the chlorophyll content also significantly decreased. Stomatal conductance (Gs), intercellular CO2 concentration (Ci), and transpiration rate (Tr) increased after flooding for 72 h. The biomass of breadfruit seedlings significantly decreased after flooding for 24 h, and showed a significant increase trend after flooding for 72 h. The chlorophyll α, b, and (α+b) contents reached the lowest values, with decreases of 23.64%, 78.17%, and 31.96%, respectively, compared to the 0-hour flooding treatment. The net photosynthetic rate (Pn), Gs, Ci, and Tr of breadfruit seedlings showed an increasing trend after flooding for 24 h, while photosynthetic parameters and antioxidant enzyme activity showed significant inhibition after flooding for 72 h and 96 h, respectively. With the increase of flooding time, the aboveground and total biomass of jackfruit seedlings increased first and then decreased, the underground and root/shoot ratio increased, the Pn, Gs, and Tr decreased, and the antioxidant enzyme activity increased. The Pn, Gs and Tr of breadfruit seedlings reached their maximum values at 24 hours of flooding and then continued to decrease, while the activities of CAT, POD, and SOD enzyme activity showed an increasing trend and then decreasing, reaching their highest values at 72 hours of flooding with the proportions of increase compared to 0 hours of flooding were 234.49%, 317.68%, and 251.88%, respectively. Correlation analysis was conducted on the waterlogging tolerance coefficients of various physiological indicators, and six waterlogging tolerance evaluation indicators were selected, including Pn, Gs, Tr, CAT, POD, and SOD. Principal component analysis showed that the variance contribution rates of the first two comprehensive indicators were 62.47% and 34.26%, respectively, with a total variance contribution rate of 96.73%, which basically included all the information of the measured indicators. Factor 1 had a significant load on Pn, Gs, and Tr, which was classified as a photosynthetic parameter. Factor 2 had a significant load on CAT, POD, and SOD, classified as antioxidant enzyme activity. Factor analysis showed that the order of waterlogging tolerance of jackfruit and breadfruit was: breadfruit-96 h＞jackfruit-72 h＞breadfruit-72 h＞jackfruit-96 h＞breadfruit-48 h＞breadfruit-24 h＞jackfruit-48 h＞jackfruit-24 h. Membership function analysis showed that the waterlogging tolerance of two Artocarpus species was from strong to weak: breadfruit-96 h＞breadfruit-24 h＞breadfruit-72 h＞jackfruit-72 h＞breadfruit-48 h＞jackfruit-96 h＞jackfruit-48 h＞jackfruit-24 h. Flooding stress significantly inhibited the growth, photosynthetic characteristics and antioxidant enzyme activities of jackfruit. Breadfruit had stronger waterlogging tolerance and response to flooding stress.【Conclusion】Flooding stress sig-nificantly inhibited the biomass, chlorophyll content, photosynthetic properties and antioxidant enzyme activities of the two Artocarpus species. Jackfruit alleviates damage caused by flooding stress through expanding root absorption, pigment regulation and increasing antioxidant enzyme activity, while breadfruit resists waterlogging by enhancing photosynthesis and antioxidant enzyme activity. In the comprehensive waterlogging tolerance score, breadfruit is better than jackfruit, which has stronger adaptive potential in short-term and medium-term flooding environment and can be used as the preferred waterlogging resistant rootstock, while jackfruit is more suitable for planting in areas with good drainage.

Breadfruit is rich in starch and pectin. The pectin can alter the digestion process of starch, however, the specific mechanism by which various soluble pectin impact starch digestion remains unclear. The effects of water-soluble pectin (WSP), chelating agent soluble pectin (CSP), and sodium carbonate soluble pectin (SSP) on the digestibility of breadfruit starch were characterized by means of RVA, FTIR, DSC, ITC, SEM and in vitro digestion. The results showed that the three types of pectin could inhibit starch digestion. The addition of CSP increased the resistant starch content from 8.18% to 18.07%,and the addition of SSP increased the slow-digested starch content from 22.55% to 35.24%. The three pectin have different ways in inhibiting starch digestion. They have different principles in inhibiting starch digestion. WSP binds with the dissolved amylose to form a membrane that covers the surface of the starch particles. However, CSP and SSP penetrate into the starch granule during the gelatinization process, blocking the pores on the starch particle surface, thus reducing the starch digestibility.

To shorten the turnaround time of molecular diagnostics, an ultrafast polymerase chain reaction (PCR) technique has been developed in recent years, which requires thermostable DNA polymerases with high kinetic properties. In the present study, Thermus aquaticus (Taq) DNA polymerase was engineered through two strategies: fusion with CL7, a thermostable nonspecific DNA-binding tag, and rational design in its N-terminal exonuclease domain. The results indicated that the CL7 tag promoted the thermostability of this polymerase dramatically, while triple substitutions, L208I/E209 K/E210R, increased its polymerase activity by approximately 5-fold. Moreover, this engineered enzyme demonstrated a strong resistance to crude plant tissue lysate up to 30% (v/v) in the PCR system. Therefore, an ultrafast qPCR system was established to detect exogenous genes in genetically modified (GM) crops with this variant. The whole detection procedure was shortened to <30 min. Moreover, the limit of detection reached approximately 0.1 copies/μL. This study established a highly simple and accurate system suitable for rapid molecular diagnosis. More importantly, this is the first report about improving the kinetic properties of Taq polymerase through rational design of its exonuclease domain.

A dual-channel detection system using Pyrococcus furiosus Argonaute (PfAgo) was developed for the quantification of single-nucleotide variants (SNVs) in genome-edited rice. The proposed approach advanced PfAgo-based detection by introducing universal guide DNA (gDNA) that simplified the system through simultaneous recognition of both wild-type and SNV targets. It achieved precise dual quantification of single-base differences without cross-interference via a rationally designed mismatch in the molecular beacons. Under the optimal conditions, the developed method exhibited exceptional sensitivity with a detection limit of 0.1% for SNV mutant, while maintaining single-nucleotide discrimination capability across various genome-edited rice variants. In blinded sample tests at 5%-10%, the system exhibited robust performance with relative standard deviation below 6% and bias within ± 8%, confirming its quantitative reliability. The proposed approach integrates high sensitivity, excellent specificity, and precise quantification, thereby providing a powerful analytical platform for traceability and quality control of genome-edited products.

The diamondback moth, Plutella xylostella, is a major pest of brassica vegetables and oilseed crops, posing a serious threat to China’s grain and oil production. RNA interference (RNAi) has been developed as an efficient strategy to control pests. In this study, the effects of RNAi on P. xylostella were evaluated by injecting two doses of synthesized dsPxvATPasea. The transcripts of PxvATPasea were widely transcribed during different developmental stages from egg to adult. They were abundantly expressed in the hindgut and Malpighian tubules, compared with other tissue types. Introduction of 800 ng dsPxvATPasea in the fourth-instar larvae greatly reduced corresponding mRNA levels by 3.1 and 1.4 times on day 2 and 3, respectively, causing 66.6% mortality and 33.4% treated larvae pupated. Silencing PxvATPasea by injecting 1200 ng dsRNA significantly decreased the expression level by 5.0 and 2.0 times on the second and third day, leading to 79.2% larval lethality and 20.8% depleted larvae pupated. Moreover, introducing 800 ng or 1200 ng dsPxvATPasea finally reduced larval fresh weight by 22.1% and 28.8%, respectively. The results indicated that the silencing efficiency of PxvATPasea worked in a dose-dependent way. Consequently, PxvATPasea is a potential molecular target gene. Our findings will facilitate the application of RNAi technology to manage P. xylostella.

The current accurate quantitative technology, such as standard curve-based quantitative real time PCR (qPCR) and digital PCR (dPCR), is time-consuming and expensive, resulting in high costs for the implementation of the GMO labeling policy. This study proposed a rapid quantitative strategy that combines the 2-Delta Delta Ctmethod with the t-test of Delta Ct values between the test samples and the reference control. This method allowed for an approximate estimation of the GMO content of test samples while identifying the GM events. GMO labeling or exempt for most samples can be determined based on the rapid estimation of GMO content and the t-test result of the Delta Ct values between the test samples and the reference control. This approach streamlines the quantitative analysis process, makes the quantification more accessible and affordable, and supports compliance with GMO labeling regulations worldwide.

This paper introduces the Silicon Valley of the United States, Singapore's Science Park, Taiwan's Hsinchu Science Park and Suzhou Industrial Park, and analyses successful experiences of these development zones in development and construction, then some common characteristics of these parks are summarized. By learning from successful experiences of development zones domestic and abroad, this paper puts forward some inspirations for the construction of China's development zones.

Artocarpus heterophyllus Lam. (jackfruit) exhibits important biological activities, with its antioxidant effects are closely associated with rich phenolic compounds. This study evaluated the total phenolic content (TPC), total flavonoid content (TFC), and phenolic components in jackfruit pulp from 21 different cultivars. 58 phenolic compounds were identified with significant variations in their quantity and content among the cultivars by using UPLC-ESI-Q-TOF-MS/MS. Notably, the cultivar “Malaysia 1” had the highest TPC and TFC, exhibited the strongest antioxidant capacity. A strong correlation between polyphenol content and antioxidant properties suggests that procyanidins B1 and p-coumaric acid are key contributors to the antioxidant activity of jackfruit pulp. Principal component analysis (PCA) revealed distinct patterns in the phenolic compound profiles across samples. These findings provide a theoretical basis for the precise cultivation of high-antioxidant varieties possible and provide a reference for the processing strategies of targeted functional jackfruit products.

Mylabris sibirica is a hypermetamorphic insect that primarily feeds on oilseed rape during the adult stage. However, the limited availability of genomic resources hinders our understanding of the gene function, medical use, and ecological adaptation in M. sibirica. Here, a high-quality chromosome-level genome of M. sibirica was generated by PacBio, Illumina, and Hi-C technologies. Its genome size was 138.45 Mb, with a scaffold N50 of 13.84 Mb and 99.85% (138.25 Mb) of the assembly anchors onto 10 pseudo-chromosomes. BUSCO analysis showed this genome assembly had a high-level completeness of 100% (n = 1,367), containing 1,358 (99.4%) single-copy BUSCOs and 8 (0.6%) duplicated BUSCOs. In addition, a total of 11,687 protein-coding genes and 35.46% (49.10 Mb) repetitive elements were identified. The high-quality genome assembly offers valuable genomic resources for exploring gene function, medical use, and ecology.

Pyrococcus furiosus Argonaute (PfAgo)-mediated nucleic acid detection is a powerful tool for molecular diagnostics, offering ultrahigh sensitivity and single-nucleotide specificity. However, existing PfAgo-based methods require multi-step cleavage processes and the use of molecular beacons, which are cumbersome and costly. Herein, we developed a novel fluorescence sensing platform (dPAFS) based on a nuclease-dead PfAgo mutant (dPfAgo) to simplify PfAgo-mediated detection system significantly. dPfAgo mutants were obtained by site-directed mutagenesis of the key catalytic site residues D628 and D558, and their functionality was confirmed through activity assay. In this sensing system, the gDNA was modified with the quenched group of black hole quencher-1 (BHQ1). Target DNA was amplified with carboxyfluorescein (FAM)-labeled primers and then precisely bound with gDNA-dPfAgo. The formed gDNA-dPfAgo-tDNA ternary complex brought the FAM donor and BHQ1 acceptor into close proximity, inducing fluorescence quenching through fluorescence resonance energy transfer. In a proof-of-concept study, dPAFS successfully genotyped genome-edited rice variants, achieving a limit of detection of 0.1 % for edited-type variant and distinguishing variants of genome-edited rice with single-nucleotide specificity. The dPAFS platform eliminates the need for molecular beacons, offering a simple, cost-efficient, and robust assay for programmable enzyme-mediated molecular diagnoses.

Epicauta gorhami is a hypermetamorphic insect that mainly forage soybeans during the adult stage. However, the lack of appropriate references hinders our studying of the gene function in E. gorhami. In this study, referring to five computational tools (Ct value, geNorm, NormFinder, BestKeeper and RefFinder), the stability of 10 housekeeping genes (GAPDH, ACT, RPL4, RPL27, α-TUB, RPS18, EF1α, RPS28, RPL13 and SOD) was assessed by qRT-PCR under three different conditions (adult ages, tissues/organs and temperatures). The findings suggested that SOD and RPS18 were the most ideal references for examine gene transcripts among diverse adult ages and at various temperatures; a pair of RPS18 and RPS28 was the most reliable genes to assess gene expressions in diverse adult tissues. Finally, the relative expression levels of EgUAP were computed after normalization RPS18 and RPS28 with across diverse adult tissues. As expected, EgUAP expression was abundant in the foregut, trachea and antenna and scarce in the midgut, hindgut and epidermis. These findings will lay a solid foundation for analyzing the gene expression of E. gorhami.

The genetically modified (GM) soybean DBN8002 has been approved for commercial planting in China. For enforcing GMO labeling policy, an event-specific real-time quantitative PCR (qPCR) method was developed to target the junction fragment between the T-DNA left border and the flanking genomic DNA, yielding a 104 base pair (bp) amplicon. This event-specific qPCR method can identify and quantify the DBN8002 event with high specificity, satisfactory linearity, and acceptable accuracy. Furthermore, the DBN8002-event specific primer/probe set was successfully transferred to a droplet digital PCR (ddPCR) platform for quantification. The quantitative results from qPCR were found to be comparable to those obtained from ddPCR, with a P-value exceeding 0.05, indicating no significant difference. The limit of detection (LOD) for both qPCR and ddPCR methods was determined to be 10 copies per reaction, while the limit of quantification (LOQ) was estimated to be 20 copies per reaction for qPCR and 40 copies per reaction for ddPCR. The collaborative validation demonstrated that the DBN8002 event-specific qPCR method had satisfactory repeatability and reproducibility. Both the event-specific qPCR and ddPCR methods are suitable for quantifying the DBN8002 content in samples. Additionally, ddPCR can be utilized for the characterization of DBN8002 reference materials.

In our previous research, the significant difference of physiochemistry properties for underutilized starches was showed between Chinese seedless breadfruit species and the other species. Based on this, the multiscale structure and digestion kinetics of Chinese seedless breadfruit of Spice and Beverage Research Institute species (SBS) and Xinglong species (XBS) was further researched. The SBS exhibited higher alpha-1,6 glycosylic bond content, free sidechain groups content, double-helix content, homogeneity, molecular weight, and V-type polymorphism, and fewer amorphous content, blocklet sizes, and a smaller semi-crystalline lamella thickness than XBS. Additionally, SBS showed higher final viscosity, pasting temperature, and gelatinization enthalpy than those of XBS. Consequently, SBS display lower rate constant (0.73 h- 1) and glycemic index (65.17) than those of XBS (0.86 h- 1 and 73.95). The anti-digestibility mechanism was revealed by the structure-digestibility relationship. It was found that resistant starch of SBS and XBS were significantly higher than those of starch from American and African species. This indicated that Chinese breadfruit starch could be considered as a good source of resistant starch, regulating glycemic index. In summary, XBS and XBS could be considered as a well source of resistant starch to make foods for preventing or improving type II diabetes or hyperlipemia.

A multi-chromatic and multi-component lateral flow immunoassay (MCMC-LFIA) was developed for simultaneous detection of CP4 EPSPS, Bt-Cry1Ab, Bt-Cry1Ac, and PAT/bar proteins in genetically modified (GM) crops. Captured antibodies specific to these exogenous proteins were separately immobilized on a nitrocellulose membrane as test zones. Multi-colored microspheres, used as visible multi-probes, were conjugated with corresponding antibodies and sprayed on the conjugate pad. The assay results can be visually interpreted within 10 min by observing the appearance of colored bands. The MCMC-LFIA demonstrated high sensitivity, with detection of limits of 7.8 ng/mL for CP4 EPSPS and 2.5 ng/mL for Bt-Cry1Ab, Bt-Cry1Ac, and PAT/bar proteins, significantly improving the performance of previously reported LFIAs. The MCMC-LFIA exhibited excellent specificity and was validated for practical use in field-based applications. The proposed MCMC-LFIA offers a rapid, sensitive, and user-friendly tool for the on-site large-scale screening of GM materials.

The structure-activity relationship and inhibitory mechanism of flavonols on alpha-glucosidase were studied by inhibition kinetics, multispectral study, and molecular docking. The flavonols of rutin, quercetin and kaempferol effectively inhibit the activity of alpha-glucosidase, among which quercetin and rutin showed the strongest and weakest inhibitory abilities, respectively. The inhibitory ability of flavonols was enhanced by hydroxylation at C3 ' of B ring, while it was weakened by diglycosylation at C3 of C ring. Remarkably, the quenching affinity and inhibitory ability of flavonols were inconsistent, which was different from the conclusions reported by some previous studies. This may be ascribed to the hydroxyl groups of C3 ' of B ring and C3 of C ring. Furthermore, three flavonols were spontaneously bound to alpha-glucosidase through hydrophobic interactions and hydrogen bonding, which caused the structure and hydrophobic microenvironment of alpha-glucosidase to change, resulting in significant inhibition of alpha-glucosidase by flavonols.