A Novel Dual-Fc Bispecific Antibody with Enhanced Fc Effector Function.

Bispecific antibodies (BsAbs) are undergoing continued development for applications in oncology and autoimmune diseases. While increasing activity by having more than one targeting arm, most BsAb engineering employs single Fc engagement as monoclonal antibodies. Here, we designed a novel immunoglobulin gamma-1 (IgG1)-derived dual-Fc BsAb containing two Fc regions and two distinct asymmetric antigen binding arms comprising a Fab arm and another VHH domain. In conjunction with the knob-into-hole technology, dual-Fc BsAbs could be produced with a high yield and good stability. We explore how Fc engineering effects on dual-Fc constructs could boost the desired therapeutic efficacy. This new format enabled simultaneous bispecific binding to corresponding antigens. Furthermore, compared to the one-Fc control molecules, dual-Fc BsAbs were shown to increase the avidity-based binding to FcγRs to result in higher ADCC and ADCP activities by potent avidity via binding to two antigens and Fc receptors. Overall, this novel BsAb format with enhanced effector functionalities provides a new option for antibody-based immunotherapy.

Strategies on biosynthesis and production of bioactive compounds in medicinal plants.

Medicinal plants are a valuable source of essential medicines and herbal products for healthcare and disease therapy. Compared with chemical synthesis and extraction, the biosynthesis of natural products is a very promising alternative for the successful conservation of medicinal plants, and its rapid development will greatly facilitate the conservation and sustainable utilization of medicinal plants. Here, we summarize the advances in strategies and methods concerning the biosynthesis and production of natural products of medicinal plants. The strategies and methods mainly include genetic engineering, plant cell culture engineering, metabolic engineering, and synthetic biology based on multiple "OMICS" technologies, with paradigms for the biosynthesis of terpenoids and alkaloids. We also highlight the biosynthetic approaches and discuss progress in the production of some valuable natural products, exemplifying compounds such as vindoline (alkaloid), artemisinin and paclitaxel (terpenoids), to illustrate the power of biotechnology in medicinal plants.

Perilla frutescens: A traditional medicine and food homologous plant.

Perilla frutescens, an annual herb of the Labiatae family, has been cultivated in China for more than 2000 years. P. frutescens is the one of the first medicinal and edible plant published by the Ministry of Health. Its leaves, stems and seeds can be used as medicine and edible food. Because of the abundant nutrients and bioactive components in this plant, P. frutescens has been studied extensively in medicine, food, health care and chemical fields with great prospects for development. This paper reviews the cultivation history, chemical compositions and pharmacological activities of P. frutescens, which provides a reference for the development and utilization of P. frutescens resources.

A chromosome-scale genome assembly of Artemisia argyi reveals unbiased subgenome evolution and key contributions of gene duplication to volatile terpenoid diversity.

Artemisia argyi Lévl. et Vant., a perennial Artemisia herb with an intense fragrance, is widely used in traditional medicine in China and many other Asian countries. Here, we present a chromosome-scale genome assembly of A. argyi comprising 3.89 Gb assembled into 17 pseudochromosomes. Phylogenetic and comparative genomic analyses revealed that A. argyi underwent a recent lineage-specific whole-genome duplication (WGD) event after divergence from Artemisia annua, resulting in two subgenomes. We deciphered the diploid ancestral genome of A. argyi, and unbiased subgenome evolution was observed. The recent WGD led to a large number of duplicated genes in the A. argyi genome. Expansion of the terpene synthase (TPS) gene family through various types of gene duplication may have greatly contributed to the diversity of volatile terpenoids in A. argyi. In particular, we identified a typical germacrene D synthase gene cluster within the expanded TPS gene family. The entire biosynthetic pathways of germacrenes, (+)-borneol, and (+)-camphor were elucidated in A. argyi. In addition, partial deletion of the amorpha-4,11-diene synthase (ADS) gene and loss of function of ADS homologs may have resulted in the lack of artemisinin production in A. argyi. Our study provides new insights into the genome evolution of Artemisia and lays a foundation for further improvement of the quality of this important medicinal plant.

Maize (Zea mays L.) responses to salt stress in terms of root anatomy, respiration and antioxidative enzyme activity.

BACKGROUND: Soil salt stress is a problem in the world, which turns into one of the main limiting factors hindering maize production. Salinity significantly affects root physiological processes in maize plants. There are few studies, however, that analyses the response of maize to salt stress in terms of the development of root anatomy and respiration. RESULTS: We found that the leaf relative water content, photosynthetic characteristics, and catalase activity exhibited a significantly decrease of salt stress treatments. However, salt stress treatments caused the superoxide dismutase activity, peroxidase activity, malondialdehyde content, Na+ uptake and translocation rate to be higher than that of control treatments. The detrimental effect of salt stress on YY7 variety was more pronounced than that of JNY658. Under salt stress, the number of root cortical aerenchyma in salt-tolerant JNY658 plants was significantly higher than that of control, as well as a larger cortical cell size and a lower root cortical cell file number, all of which help to maintain higher biomass. The total respiration rate of two varieties exposed to salt stress was lower than that of control treatment, while the alternate oxidative respiration rate was higher, and the root response of JNY658 plants was significant. Under salt stress, the roots net Na+ and K+ efflux rates of two varieties were higher than those of the control treatment, where the strength of net Na+ efflux rate from the roots of JNY658 plants and the net K+ efflux rate from roots of YY7 plants was remarkable. The increase in efflux rates reduced the Na+ toxicity of the root and helped to maintain its ion balance. CONCLUSION: These results demonstrated that salt-tolerant maize varieties incur a relatively low metabolic cost required to establish a higher root cortical aerenchyma, larger cortical cell size and lower root cortical cell file number, significantly reduced the total respiration rate, and that it also increased the alternate oxidative respiration rate, thereby counteracting the detrimental effect of oxidative damage on root respiration of root growth. In addition, Na+ uptake on the root surface decreased, the translocation of Na+ to the rest of the plant was constrained and the level of Na+ accumulation in leaves significantly reduced under salt stress, thus preempting salt-stress induced impediments to the formation of shoot biomass.

Effects of Hydroxypropyl Methylcellulose on Physicochemical Properties and Microstructure of κ-Carrageenan Film.

We investigated the effects of different proportions of hydroxypropyl methylcellulose (HPMC) on the properties of κ-carrageenan film. Biodegradable κ-carrageenan/HPMC films (κCHM film) were prepared by the solution casting method and their physicochemical properties were evaluated. The results show that the addition of HPMC enhanced oxygen barrier capacity, mechanical properties (tensile strength and elongation at break) and thermal stability. Notably, when the addition of HPMC increased to 6% of κ-carrageenan (w:w), the κCHM-6 film not only effectively improved water resistance, including lower water solubility, water vapor permeability and higher water contact angle, but also made the structure of the κCHM-6 film more compact. Moreover, rheological measurement and atomic force microscopy characterization showed that κ-carrageenan had suitable compatibility with HPMC. Attenuated total reflection-Fourier transform infrared spectroscopy analysis further confirmed the enhancement of hydrogen bond interactions. This finding could contribute to promoting the potential application of κCHM film in food packaging.

CRISPR-Cas gene editing technology and its application prospect in medicinal plants.

The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas gene editing technology has opened a new era of genome interrogation and genome engineering because of its ease operation and high efficiency. An increasing number of plant species have been subjected to site-directed gene editing through this technology. However, the application of CRISPR-Cas technology to medicinal plants is still in the early stages. Here, we review the research history, structural characteristics, working mechanism and the latest derivatives of CRISPR-Cas technology, and discussed their application in medicinal plants for the first time. Furthermore, we creatively put forward the development direction of CRISPR technology applied to medicinal plant gene editing. The aim is to provide a reference for the application of this technology to genome functional studies, synthetic biology, genetic improvement, and germplasm innovation of medicinal plants. CRISPR-Cas is expected to revolutionize medicinal plant biotechnology in the near future.

6-Benzyladenine increasing subsequent waterlogging-induced waterlogging tolerance of summer maize by increasing hormone signal transduction.

Summer maize is frequently subjected to waterlogging damage because of increased and variable rainfall during the growing season. The application of 6-benzyladenine (6-BA) can effectively mitigate the waterlogging effects on plant growth and increase the grain yield of waterlogged summer maize. However, the mechanisms underlying this process and the involvement of 6-BA in relevant signal transduction pathways remain unclear. In this study, we explored the effects of 6-BA on waterlogged summer maize using a phosphoproteomic technique to better understand the mechanism by which summer maize growth improves following waterlogging. Application of 6-BA inhibited the waterlogging-induced increase in abscisic acid (ABA) content and increased the phosphorylation levels of proteins involved in ABA signaling; accordingly, stomatal responsiveness to exogenous ABA increased. In addition, the application of 6-BA had a long-term effect on signal transduction pathways and contributed to rapid responses to subsequent stresses. Plants primed with 6-BA accumulated more ethylene and jasmonic acid in response to subsequent waterlogging; accordingly, leaf SPAD, antioxidase activity, and root traits improved by 6-BA priming. These results suggest that the effects of 6-BA on hormone signal transduction pathways are anamnestic, which enables plants to show faster or stronger defense responses to stress.

Recent Advances in Postharvest Irradiation Preservation Technology of Edible Fungi: A Review.

Edible fungi have high edible, medicinal and economic value. Rapid development of the edible fungi industry can meet people's consumption demands. However, due to lack of suitable preservation technology after harvest, edible fungi are susceptible to mechanical damage, microbial infection, and discoloration, which could affect the quality and shelf life of fresh edible fungi. Many techniques have been developed to extend the postharvest storage time of fresh edible fungi and irradiation technology has been proven to be one of the potential technologies. This review summarizes the internal and external factors affecting the postharvest quality deterioration of edible fungi, introduces the types of irradiation preservation technology and describes comprehensive advances in the effects of irradiation on shelf life, microbiology, organoleptic qualities, nutritional qualities (proteins, fats, sugars and vitamins) and enzymatic activities of edible fungi from different regions and of different species worldwide. This review uncovers that the postharvest quality decay of edible fungi is a complex process. The irradiation preservation of edible fungi is affected not only by the edible fungus itself and the storage environment but also by the radiation type, radiation dose and radiation source conditions. Future studies need to consider the combined application of irradiation and other novel technologies to further improve the preservation effect of edible fungi, in particular in the area of irradiation's influence on the flavor of edible fungus.

The reference genome of Miscanthus floridulus illuminates the evolution of Saccharinae.

Miscanthus, a member of the Saccharinae subtribe that includes sorghum and sugarcane, has been widely studied as a feedstock for cellulosic biofuel production. Here, we report the sequencing and assembly of the Miscanthus floridulus genome by the integration of PacBio sequencing and Hi-C mapping, resulting in a chromosome-scale, high-quality reference genome of the genus Miscanthus. Comparisons among Saccharinae genomes suggest that Sorghum split first from the common ancestor of Saccharum and Miscanthus, which subsequently diverged from each other, with two successive whole-genome duplication events occurring independently in the Saccharum genus and one whole-genome duplication occurring in the Miscanthus genus. Fusion of two chromosomes occurred during rediploidization in M. floridulus and no significant subgenome dominance was observed. A survey of cellulose synthases (CesA) in M. floridulus revealed quite high expression of most CesA genes in growing stems, which is in agreement with the high cellulose content of this species. Resequencing and comparisons of 75 Miscanthus accessions suggest that M. lutarioriparius is genetically close to M. sacchariflorus and that M. floridulus is more distantly related to other species and is more genetically diverse. This study provides a valuable genomic resource for molecular breeding and improvement of Miscanthus and Saccharinae crops.

Narrowing Yield Gaps and Enhancing Nitrogen Utilization for Summer Maize (Zea mays L) by Combining the Effects of Varying Nitrogen Fertilizer Input and Planting Density in DSSAT Simulations.

In China, the most common grain crop is maize (Zea mays). The increasing pressure to meet the food demands of its growing population has pushed Chinese maize farmers toward an excessive use of chemical fertilizers, a practice which ultimately leads to a massive waste of resources and widespread environmental pollution. As a result, increasing the yield and improving the nitrogen (N) use efficiency of maize has become a critical issue for agriculture in China. This study, which analyzes the combined data from a simulation carried out using the Decision Support System for Agrotechnology Transfer (DSSAT), a field experiment, and a household survey, explored the effectiveness of several approaches aimed at narrowing the maize yield gap and improving the N utilization efficiency in the Huang-Huai-Hai Plain (HHHP), the most important area for the production of summer maize in China. The various approaches we studied deploy different methods for the integrated management of N fertilizer input and the planting density. The study produced the following results: (1) For the simulated and actual maize yields, the root mean square error (RMSE), the normalized root mean squared errors (NRMSE) and the index of agreement (d) were 1,171 (kg ha-1), 12% and 0.84, respectively. These results show that the model is viable for the experiment included in the study; (2) The potential yield was 15.58 t ha-1, and the yields achieved by the super-high-yield cultivation pattern (SH), the optimized nutrient and density management pattern (ONM), the simulated farmer's practice cultivation pattern (FP) and actual farmer's practice (AFP) were 11.43, 11.06, 10.33, and 7.95 t ha-1, respectively. The yield gaps associated with the different yield levels were large; (3) For summer maize, the high yield and a high N partial factor productivity (NPFP) was found when applying a planting density of 9 plants m-2 and an N application amount of 246 kg ha-1. These results suggest that the maximum yield that can actually be achieved by optimizing the N application and planting density is less than 73% of the potential yield. This implies in turn that in order to further narrow the observed yield gaps, other factors, such as irrigation, sowing dates and pest control need to be considered.

Integrating density and fertilizer management to optimize the accumulation, remobilization, and distribution of biomass and nutrients in summer maize.

Improved the utilization of fertilizer while maintaining the increased of grain yield was the focus of Chinese researchers. Nutrient uptake, distribution, and remobilization are important factors affecting the fertilizer utilization and grain yield of maize. This study aimed to provide a theoretical and practical basis for science-based, high-yielding, and high-efficiency cultivation practices by examining differences in biomass and nutrient uptake, distribution, and remobilization characteristics under three cultivation patterns. We set 12 treatments as follows: super high-yielding cultivation pattern (SH), optimized nutrient management cultivation pattern (ONM), local farmer's practice cultivation pattern (FP), and a series of nutrient omission plots, which excluded nitrogen (N), phosphorus (P), or potassium (K) from the three patterns. The results demonstrated that SH and ONM increased the yield and actual harvested ears by 35.4, 20.7 and by 20.2, 17.6%, respectively. Compared with the FP, SH and ONM increased biomass, N, P, and K accumulation at silking (R1 stage) by 24.4, 31.2, 39.4, and 34.8%, and by 21.7, 22.2, 31.7, and 34.8%, respectively. SH and ONM significantly increased biomass and nutrient distribution to the grains. ONM significantly increased N use efficiency. P and K use efficiency under the ONM pattern was significantly higher than under SH, but was lower than under the FP pattern over two years. This research demonstrates that ONM may significantly reduce fertilizer rates, effectively improve the nutrient remobilization efficiency and uptake at post-silking without negatively affecting grain yield, thereby increasing N use efficiency.

Comparative proteomic analysis reveals that exogenous 6-benzyladenine (6-BA) improves the defense system activity of waterlogged summer maize.

BACKGROUND: Exogenous 6-benzyladenine (6-BA) could improve leaf defense system activity. In order to better understand the regulation mechanism of exogenous 6-benzyladenine (6-BA) on waterlogged summer maize, three treatments including control (CK), waterlogging at the third leaf stage for 6 days (V3-6), and application of 100 mg dm- 3 6-BA after waterlogging for 6 days (V3-6-B), were employed using summer maize hybrid DengHai 605 (DH605) as the experimental material. We used a labeling liquid chromatography-based quantitative proteomics approach with tandem mass tags to determine the changes in leaf protein abundance level at the tasseling stage. RESULTS: Waterlogging significantly hindered plant growth and decreased the activities of SOD, POD and CAT. In addition, the activity of LOX was significantly increased after waterlogging. As a result, the content of MDA and H2O2 was significantly increased which incurred serious damages on cell membrane and cellular metabolism of summer maize. And, the leaf emergence rate, plant height and grain yield were significantly decreased by waterlogging. However, application of 6-BA effectively mitigated these adverse effects induced by waterlogging. Compared with V3-6, SOD, POD and CAT activity of V3-6-B were increased by 6.9, 12.4, and 18.5%, LOX were decreased by 13.6%. As a consequence, the contents of MDA and H2O2 in V3-6-B were decreased by 22.1 and 17.2%, respectively, compared to that of V3-6. In addition, the leaf emergence rate, plant height and grain yield were significantly increased by application of 6-BA. Based on proteomics profiling, the proteins involved in protein metabolism, ROS scavenging and fatty acid metabolism were significantly regulated by 6-BA, which suggested that application of 6-BA exaggerated the defensive response of summer maize at proteomic level. CONCLUSIONS: These results demonstrated that 6-BA had contrastive effects on waterlogged summer maize. By regulating key proteins related to ROS scavenging and fatty acid metabolism, 6-BA effectively increased the defense system activity of waterlogged summer maize, then balanced the protein metabolism and improved the plant physiological traits and grain yield.

Maize Canopy Apparent Photosynthesis and 13C-Photosynthate Reallocation in Response to Different Density and N Rate Combinations.

Appropriate planting density and nitrogen (N) supply are critical factors optimizing yield in crop cultivation. To advance the knowledge of maize plants under different density and N rate combinations, responses of canopy apparent photosynthesis (CAP), and assimilate redistribution characters (by 13CO2 stable isotope tracing) were investigated. In this study, two maize varieties DH618 and DH605 were grown at various planting densities (6.75, 8.25, 9.75, and 11.25 pl m-2) and N application rates (0, 180, 270, 360, and 540 kg ha-1) during 2013-2015. Maize grain yield (GY) was maximized at a density of 9.75 pl m-2 with 180-360 kg ha-1 N during the three study years. Maize GY, biomass, CAP, leaf area index (LAI), and 13C-photosynthate reallocation all responded more intensively to density than N rate, but the N response differed between varieties. We established links among CAP, LAI and biomass, and GY and kernel number per unit area (KNA). CAP depended on high LAI and enzyme activities for photosynthesis, yet both N deficiency and N excess had inhibitory effects. Besides, relations between 13C-photosynthate reallocation and yield components were executed. High density increased the 13C-photosynthate distribution in vegetative organs but reduced the allocation in ear, while N supply moderated the response. Based on our results, maize plants with greater CAP, more 13C-photosynthate distribution to ears, and less 13C-photosynthate distribution to stems under different density and N rate combinations could improve KNA and achieve a greater GY consequently.

Plant Density and Nitrogen Supply Affect the Grain-Filling Parameters of Maize Kernels Located in Different Ear Positions.

Although yield output of maize (Zea mays L.) has improved markedly over the last century, procedures for improving the grain-filling process remain elusive. Our aim in this study was to relate grain-filling variation in maize (including kernels in apical and middle positions in the ears) to plant density and nitrogen (N) application rate using a crossed experimental design. We also investigated changes in zeatin riboside (ZR), indole-3-acetic acid (IAA), abscisic acid (ABA), and gibberellic acid (GA) in the kernels during the grain-filling period. Two high-yield maize varieties cultivated extensively in China were field grown under normal (67,500 pl ha-1) and high (97,500 pl ha-1) densities, and supplied with low, normal and high (0, 180, and 360 kg N ha-1) concentrations of N. Kernel weight (KW), the maximum grain-filling rate (Gmax), the average grain-filling rate (Gave), and the kernel weight increment achieving Gmax (Wmax) were all significantly depressed under high density (HD) conditions, but increased N supply partially offset the losses. The apical kernels were more sensitive to density and N application rate than middle kernels. Correlation analysis indicated that plant density and N rate affected KW mainly by influencing the grain-filling rate. Variation in ZR, IAA, and ABA content tracked the variation in KW, but variation in GA content did not. Furthermore, the grain-filling parameters (closely related to TKW) had strong canonical correlation with the content of all hormones across the filling period and ZR content had the strongest relationship. Based on our study, high N supply is beneficial to optimize grain-filling parameters and improve KW of maize kernels under crowded condition.

Physiological and proteome studies of maize (Zea mays L.) in response to leaf removal under high plant density.

BACKGROUND: Under high plant density, intensifying competition among individual plants led to overconsumption of energy and nutrients and resulted in an almost dark condition in the lower strata of the canopy, which suppressed the photosynthetic potential of the shaded leaves. Leaf removal could help to ameliorate this problem and increase crop yields. To reveal the mechanism of leaf removal in maize, tandem mass tags label-based quantitative analysis coupled with liquid chromatography-tandem mass spectrometry were used to capture the differential protein expression profiles of maize subjected to the removal of the two uppermost leaves (S2), the four uppermost leaves (S4), and with no leaf removal as control (S0). RESULTS: Excising leaves strengthened the light transmission rate of the canopy and increased the content of malondialdehyde, whereas decreased the activities of superoxide dismutase and peroxidase. Two leaves removal increased the photosynthetic capacity of ear leaves and the grain yield significantly, whereas S4 decreased the yield markedly. Besides, 239 up-accumulated proteins and 99 down-accumulated proteins were identified between S2 and S0, which were strongly enriched into 30 and 23 functional groups; 71 increased proteins and 42 decreased proteins were identified between S4 and S0, which were strongly enriched into 22 and 23 functional groups, for increased and decreased proteins, respectively. CONCLUSIONS: Different defoliation levels had contrastive effects on maize. The canopy light transmission rate was strengthened and proteins related to photosynthetic electron-transfer reaction were up-regulated significantly for treatment S2, which improved the leaf photosynthetic capacity, and obtained a higher grain yield consequently. In contrast, S4 decreased the grain yield and increased the expressions of proteins and genes associated with fatty acid metabolism. Besides, both S2 and S4 exaggerated the defensive response of maize in physiological and proteomic level. Although further studies are required, the results in our study provide new insights to the further improvement in maize grain yield by leaf removal.

Grain development and endogenous hormones in summer maize (Zea mays L.) submitted to different light conditions.

Low light is a type of abiotic stress that seriously affects plant growth and production efficiency. We investigated the response mechanisms of summer maize to low light by measuring the changes in endogenous hormones in the grains and during grain filling in summer maize at different light intensities to provide a theoretical basis for the production and management of summer maize under light stress. We applied different light treatments in a field experiment as follows: S, shading from tassel stage (VT) to maturity stage (R6); CK, natural lighting in the field; and L, increasing light from VT to R6. The shading level was 60%, and the maximum illumination intensity of the increasing light treatment on cloudy days was 1600-1800 µmol m-2 s-1. Compared with the control, shading significantly increased the grain abscisic acid (ABA) content at 5-20 days after pollination and decreased the indole acetic acid (IAA), zeatin riboside (ZR), and gibberellin (GA) contents (P < 0.05). The grain-filling rate decreased under shading conditions. Meanwhile, the grain volume, grain weight, and yield all decreased; the yields in 2013 and 2014 decreased by 61 and 60%, respectively. The grain IAA, ZR, and GA contents were increased by increasing light. The grain ABA content at 5-20 days after pollination did not significantly differ from that of CK (P < 0.05). After 20 days after pollination, the ABA content decreased, the grain-filling rate and the filling duration increased, and the yield increased. However, shading after anthesis increased the grain ABA content and reduced the IAA, ZR, and GA contents. Grain growth and development were inhibited, and the yield decreased. The grain ABA content decreased; the IAA, ZR, and GA contents increased; and the yield increased after increasing light. The results indicate that different light intensities regulated the levels of grains endogenous hormones, which influenced the grain-filling rate and duration, and consequently, regulated grain weight and yield.

Response of Summer Maize Photosynthate Accumulation and Distribution to Shading Stress Assessed by Using 13CO2 Stable Isotope Tracer in the Field.

Maize is one of the most important crops globally that provides food, feed, and bioenergy. However, shading stress threatens maize production. In this study, we investigated the effects of shading on photosynthate accumulation and distribution of summer maize in the field. Zhengdan958 (ZD958) and Denghai 605 (DH605) were used as experimental materials in a field experiment running from 2013 to 2015. Shading treatments were applied over different growth stages: from the tassel stage (VT) to physiological maturity (R6) (S1), from the six-leaf stage (V6) to VT (S2), and from emergence stage (VE) to R6 (S3). The effects of shading on plant photosynthesis, photosynthate accumulation and distribution, and yield were evaluated in comparison to ambient sunlight. Shading significantly decreased the leaf area, SPAD value, net photosynthetic rate, dry matter accumulation, and grain yield. During the 3-year experimental period, grain yields of ZD958 and DH605 were reduced by 83.4%, 34.2%, 53.1% and 79.3%, 24.2%, 57.6% as compared to the CK by treatments S3, S2, and S1, respectively. 13CO2 stable isotope tracing revealed that shading differentially affected the photosynthate transfer rate in different stages; photosynthates were transferred from top to bottom plant parts, in the order control > S2 > S1 > S3. We conclude that shading clearly disrupted photosynthate metabolism, and reduced the photosynthate accumulation in the grain, resulting in a yield reduction.