Embedded Bioprinting of Tissue-like Structures Using κ-Carrageenan Sub-Microgel Medium.

Embedded three-dimensional (3D) bioprinting utilizing a granular hydrogel supporting bath has emerged as a critical technique for creating biomimetic scaffolds. However, engineering a suitable gel suspension medium that balances precise bioink deposition with cell viability and function presents multiple challenges, particularly in achieving the desired viscoelastic properties. Here, a novel κ-carrageenan gel supporting bath is fabricated through an easy-to-operate mechanical grinding process, producing homogeneous sub-microscale particles. These sub-microgels exhibit typical Bingham flow behavior with small yield stress and rapid shear-thinning properties, which facilitate the smooth deposition of bioinks. Moreover, the reversible gel-sol transition and self-healing capabilities of the κ-carrageenan microgel network ensure the structural integrity of printed constructs, enabling the creation of complex, multi-layered tissue structures with defined architectural features. Post-printing, the κ-carrageenan sub-microgels can be easily removed by a simple phosphate-buffered saline wash. Further bioprinting with cell-laden bioinks demonstrates that cells within the biomimetic constructs have a high viability of 92% and quickly extend pseudopodia, as well as maintain robust proliferation, indicating the potential of this bioprinting strategy for tissue and organ fabrication. In summary, this novel κ-carrageenan sub-microgel medium emerges as a promising avenue for embedded bioprinting of exceptional quality, bearing profound implications for the in vitro development of engineered tissues and organs.

Bioprinted biomimetic hydrogel matrices guiding stem cell aggregates for enhanced chondrogenesis and cartilage regeneration.

Articular cartilage tissue has limited self-repair capabilities, with damage frequently progressing to irreversible degeneration. Engineered tissues constructed through bioprinting and embedded with stem cell aggregates offer promising therapeutic alternatives. Aggregates of bone marrow mesenchymal stromal cells (BMSCs) demonstrate enhanced and more rapid chondrogenic differentiation than isolated cells, thus facilitating cartilage repair. However, it remains a key challenge to precisely control biochemical microenvironments to regulate cellular adhesion and cohesion within bioprinted matrices simultaneously. Herein, this work reports a bioprintable hydrogel matrix with high cellular adhesion and aggregation properties for cartilage repair. The hydrogel comprises an enhanced cell-adhesive gelatin methacrylate and a cell-cohesive chitosan methacrylate (CHMA), both of which are subjected to photo-initiated crosslinking. By precisely adjusting the CHMA content, the mechanical stability and biochemical cues of the hydrogels are finely tuned to promote cellular aggregation, chondrogenic differentiation and cartilage repair implantation. Multi-layer constructs encapsulated with BMSCs, with high cell viability reaching 91.1%, are bioprinted and photo-crosslinked to support chondrogenic differentiation for 21 days. BMSCs rapidly form aggregates and display efficient chondrogenic differentiation both on the hydrogels and within bioprinted constructs, as evidenced by the upregulated expression of Sox9, Aggrecan and Collagen 2a1 genes, along with high protein levels. Transplantation of these BMSC-laden bioprinted hydrogels into cartilaginous defects demonstrates effective hyaline cartilage repair. Overall, this cell-responsive hydrogel scaffold holds immense promise for applications in cartilage tissue engineering.

Phase and detuning control of the unidirectional reflection amplification based on the broken spatial symmetry.

In order to achieve the tunable unidirectional reflection amplification in a uniform atomic medium that is of vital importance to design high-quality nonreciprocal photonic devices, we propose a coherent closed three-level Δ-type atomic system by applying a microwave field, and a strong coupling field of linear variation along the x direction to control a probe field. In our scheme, the linearly increased coupling field destroys the spatial symmetry of probe susceptibility and effectively suppresses the reflection of one side; the microwave field constructs closed loop transitions to amplify the probe field and causes phase changes. The numerical simulation indicates that the unidirectional reflection amplification is sensitive to the relative phase ϕ and the coupling detuning Δc. Our results will open a new route toward harnessing optical non-reciprocity, which can provide more convenience and possibilities in the experimental realization.

Chemoselectivity of the CuAAC/Ring Cleavage/Cyclization Reaction between Enaminones and α-Acylketenimine.

Ketenimines represent an important class of reactive species, useful synthetic intermediates, and synthons. However, in general, ketenimines preferentially undergoes nucleophilic addition reactions with hydroxyl and amino groups, and carbon functional groups remain a less studied subset of such systems. Herein, we develop a straightforward syntheses of pyridin-4(1H)-imines that is achieved by cyclization of a reacting enaminone unit with α-acylketenimine which is generated from the reactions of sulfonyl azides and terminal ynones in situ (CuAAC/Ring cleavage reaction). The cascade process preferentially starts with the nucleophilic α-C of the enaminone unit instead of an amino group, attacking the electron-deficient central carbon of ketenimine, and the chemoselectivity unconventional products pyridin-4(1H)-imines were formed by intramolecular cyclization.

Spatial susceptibility modulation and controlled unidirectional reflection amplification via four-wave mixing.

Control of unidirectional light propagation is of paramount importantance to optical signal processing and optical communication. Especially, the amplified optical signal can isolate noise well that may provide more applications. In this work, we propose a dynamically modulated regime to realize unidirectional reflection amplification in a short and dense uniform atomic medium, and all atoms are driven into four-level double-Λ type by two coupling fields with linearly varied intensities along x direction and two weak probe fields. Based on four-wave mixing resonance and the broken spatial symmetry, the complete nonreciprocal reflection (unidirectional reflection) can be amplified with reflectivity more than 2.0, even to 6.0. In addition, the width, height, and position of the unidirectional reflection bands can be tunable. Thus, our regime is feasible and may inspire further applications in all-optical networks that require controllable unidirectional light amplification.

Comprehensive assessment of base excision repair (BER)-related lncRNAs as prognostic and functional biomarkers in lung adenocarcinoma: implications for personalized therapeutics and immunomodulation.

BACKGROUND: Lung adenocarcinoma (LUAD) is the most prevalent subtype of lung cancer, and comprehending its molecular mechanisms is pivotal for advancing treatment efficacy. This study aims to explore the prognostic and functional significance of base excision repair (BER)-related long non-coding RNAs (BERLncs) in LUAD. METHODS: A risk score model for BERLncs was developed using the least absolute shrinkage and selection operator regression and Cox regression analysis. Model validation and prognostic evaluation were performed using Kaplan-Meier and receiver-operating characteristic curve analyses. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were conducted to elucidate the potential biological functions of BERLncs. Comparative analyses were carried out to investigate disparities in tumor mutation burden (TMB), immune infiltration, tumor immune dysfunction and exclusion (TIDE) score, chemosensitivity, and immune checkpoint gene expression between the two risk groups. RESULTS: A predictive risk score model comprising 19 BERLncs was successfully developed. Patients were divided into high-risk and low-risk groups according to the median risk score. The high-risk subgroup exhibited significantly inferior overall survival. Functional enrichment analysis revealed pathways associated with lung cancer development, notably the neuroactive ligand-receptor interaction pathway. High-risk patients demonstrated elevated TMB, diminished TIDE scores, and an immunosuppressive tumor microenvironment, while low-risk patients displayed potential benefits from immunotherapy. Additionally, the risk model identified potential anticancer agents. CONCLUSION: The risk score model based on BERLncs shows promise as a prognostic biomarker for LUAD patients, providing valuable insights for clinical decision-making, therapeutic strategies, and understanding of underlying biological mechanisms.

Synthesis of Pyridin-1(2H)-ylacrylates and the Effects of Different Functional Groups on Their Fluorescence.

While fluorescent organic materials have many potential as well as proven applications and so have attracted significant attention, pyridine-olefin conjugates remain a less studied subset of such systems. Herein, therefore, we report on the development of the straightforward syntheses of pyridin-1(2H)-ylacrylates and the outcomes of a study of the effects of substituents on their fluorescent properties. Such compounds were prepared using a simple, metal-free and three-component coupling reaction involving 2-aminopyridines, sulfonyl azides and propiolates. The fluorescent properties of the ensuing products are significantly affected by the positions of substituents on the cyclic framework, with those located in central positions having the greatest impact. Electron-withdrawing groups tend to induce blue shifts while electron-donating ones cause red shifts. This work highlights the capacity that the micro-modification of fluorescent materials provides for fine-tuning their properties such that they may be usefully applied to, for example, the study of luminescent materials.

Mild and efficient synthesis of benzothiazolopyrimidine derivatives via CuAAC/ring cleavage/cyclization reaction.

An operationally mild and efficient synthesis of benzothiazolopyrimidine is achieved by a three-component reaction of 2-aminebenzo[d]thiazoles, sulfonyl azides and terminal ynones. This cascade process involved a CuAAC/ring cleavage/cyclization reaction. Particularly, most of the benzothiazolopyrimidine derivatives could be isolated by filtration without further purification.

Copper-catalyzed three-component reaction to synthesize polysubstituted imidazo[1,2-a]pyridines.

An efficient three-component one-pot and operationally simple cascade of 2-aminopyridines with sulfonyl azides and terminal ynones is reported, providing a variety of polysubstituted imidazo[1,2-a]pyridine derivatives in moderate to excellent yields. In particular, the reaction goes a through CuAAC/ring-cleavage process and forms a highly active intermediate α-acyl-N-sulfonyl ketenimine with base free.

Genetic diversity of the cultivated Salvia miltiorrhiza populations revealed by four intergenic spacers.

For better understanding the genetic diversity and phylogeny of the cultivated Salvia miltiorrhiza populations, four intergenic spacer sequences, ETS, psbA-trnH, trnL-trnF, and ycf1-rps15 of the 40 populations collected from China were Polymerase Chain Reaction (PCR) amplified, analyzed both individually and in combination. Haplotype diversity analysis showed that the cultivated S. miltiorrhiza populations had a very rich genetic diversity and an excellent capacity to resist environmental pressure. The best-fit nucleotide substitution models for ETS, psbA-trnH, trnL-trnF, ycf1-rps15, and their combined sequences were HKY+I, T92, T92, T92+G, and T92+G, respectively; the nucleotide conversion frequency in the combined sequences was lower than the transversion, and the relatively high nucleotide substitution frequencies suggests its high genetic variability. Neutral tests showed that the spacer sequences of the populations conform with the neutral evolution model, and there has been no current expansion events occurred. Phylogeny analyses based on both the individual and the combined sequences showed that the 40 populations were clustered in two clades with a very similar topological structure. The discrimination rate of the combined sequence marker is significantly increased to 52.5% (21 populations) over the highest 35% (13 populations) by the single marker of ETS, though still inadequate but a big step forward. Further exploration of more DNA markers is needed. This study for the first time revealed the rich genetic diversity and phylogeny of the currently cultivated S. miltiorrhiza populations in China and provides novel alternative molecular markers for the genetic identification and resources evaluation of the cultivated S. miltiorrhiza populations.

Overexpression of LpCPC from Lilium pumilum confers saline-alkali stress (NaHCO3) resistance.

Lilium Pumilum with wide distribution is highly tolerant to salinity. The blue copper protein LpCPC (Lilium pumilum Cucumber Peeling Cupredoxin) gene was cloned from Lilium pumilum, which has the conserved regions of type I copper protein. Moreover, LpCPC has the closest relation to CPC from Actinidia chinensis using DNAMAN software and MEGA7 software. qRT-PCR indicated that LpCPC expression was higher in root and bulb of Lilium pumilum, and the expression of the LpCPC gene increased and reached the highest level at 12 h in bulbs under 20 mM NaHCO3. The transgenic yeast was more tolerant compared with the control under NaHCO3 stress. Compared with the wild type, overexpressing plants indicated a relatively lower degree of wilting. In addition, the chlorophyll content, soluble phenol content, and lignin content of overexpressing lines were higher than that of wild-type, whereas the relative conductivity of overexpressing plants was significantly lower than that of wild-type plants. Expression of essential genes including NHX1 and SOS1 in salt stress response pathways are steadily higher in overexpression tobacco than that in wild-types. Transgenic lines had much higher levels of CCR1 and CAD, which are involved in lignin production, compared with wild-type lines. The yeast two-hybrid technique was applied to screen probable interacting proteins interacting with LpCPC. Eight proteins interacted with LpCPC were screened, and five of which were demonstrated to be associated with plant salinity resistance. Overall, the role of gene LpCPC is mediating molecule responses in increasing saline-alkali stress resistance, indicating that it is an essential gene to enhance salt tolerance in Lilium pumilum.

Functional verification and screening of protein interacting with the slPHB3.

slPHB3 was cloned from Salix linearistipularis, the amino acid sequence blast and phylogenetic tree analysis showed that slPHB3 has the most similarity with PHB3 from Populus trichocarpa using DNAMAN software and MEGA7 software. RT-qPCR results confirmed that the expression of slPHB3 was induced obviously under stress treatments. The growth of recombinant yeast cells was better than that of the control group under the stress treatment, indicating that slPHB3 may be involved in the stress response of yeast cells. The transgenic tobacco was treated with different concentrations of NaCl, NaHCO3 and H2O2, fresh weigh of overexpression tobacco were heavier than wild-types. The results showed that transgenic tobacco was more tolerant to salt and oxidation than wild-type tobacco. Expression of important genes including NHX1 and SOS1 in salt stress response pathways are steadily higher in overexpression tobacco than that in wild-types. We identified 17 proteins interacting with slPHB3 by yeast two-hybrid technique, most of these proteins were relation to the stresses. The salt tolerance of slPHB3 expressing yeast and slPHB3 overexpressing plants were better than that of the control. Ten stress-related proteins may interact with slPHB3, which preliminarily indicated that slPHB3 had a certain response relationship with salt stress. The study of slPHB3 under abiotic stress can improve our understanding of PHB3 gene function.

Occurrence and distribution of antibiotics in groundwater, surface water, and sediment in Xiong'an New Area, China, and their relationship with antibiotic resistance genes.

The emergence and pollution of antibiotics in surface water in various regions have drawn widespread concern because of the harm to aquatic ecosystems and human health. In this study, we aim to first investigate contamination and ecological risks of 39 antibiotics in Xiong'an New Area (XANA), China, and then illuminate relative abundances of antibiotic resistance genes (ARGs) and their correlations with antibiotics. The sum of antibiotic concentrations in the water circulation system, including surface water, groundwater, and sediment was 12.71-260.56 ng/L, ND-196.12 ng/L, and 38.03-406.31 ng/g, respectively. In surface water and sediment, cephalosporins and quinolones were the primary antibiotics, accounting for 45% and 16% of the total antibiotic concentrations in surface water and for 62% and 32% of the total antibiotic concentrations in sediment; this suggests a significant interaction between the two media. The antibiotic concentration was the highest in shallow groundwater at depths of <50 m (mean concentration of 79.22 ± 56.46 ng/L), indicating that surface water was a possible source of antibiotic contamination in groundwater. AMX presented the highest risk in both surface and groundwater and should be controlled as a priority. Moreover, the selection pressure of antibiotics on ARGs was discovered in the sediment in XANA, because the enrichment of sulA was significantly correlated with spiramycin and lincomycin and the enrichment of blaOXA-1 was significantly correlated with roxithromycin, ciprofloxacin, ofloxacin, and sulfapyridine. Thus, our investigation revealed potential antibiotic contamination in multiple environmental media in XANA, which should be addressed to prevent more serious pollution.

Two interaction proteins between AtPHB6 and AtSOT12 regulate plant salt resistance through ROS signaling.

In the past, the PHB gene function was mainly focused on anti-cell proliferation and antitumor effects. But the molecular mechanism of the PHB gene regarding saline and oxidative stresses is unclear. To study the role of AtPHB6 in salt and oxidative stress, AtPHB6 was cloned from A. thaliana. Bioinformatics analysis showed that AtPHB6 was closely related to AtPHB1 and AtPHB2, which are both type II PHB. RT-qPCR results indicated that the AtPHB6 in the leaves and roots of A. thaliana was obviously induced under different stress treatments. AtPHB6-overexpressing plants were larger and more lush than wild-type and mutant plants when placed under stress treatments during seed germination. The root length and fresh weight of AtPHB6 transgenic plants showed the best resistance compared to wild-type plants under different treatments, in contrast, the AtPHB6 mutants had the worst resistance during the seedling stage. AtSOT12 was an interacting protein of AtPHB6, which screened by yeast two-hybrid system. The interaction between the two proteins were further confirmed using in vitro pull-down experiments and in vivo BiFC experiments. Subcellular localization showed both AtPHB6 and AtSOT12 protein expressed in the nucleus and cytoplasm. The H2O2 content in both the transgenic AtPHB6 and AtSOT12 plants were lower than that in the wild type under stresses. Thus, AtPHB6 increased plant resistance to salt stress and interacted with the AtSOT12 protein.

Progress and prospect of single-molecular ClpX ATPase researching system-a mini-review.

ClpXP in Escherichia coli is a proteasome degrading protein substrates. It consists of one hexamer of ATPase (ClpX) and two heptamers of peptidase (ClpP). The ClpX binds ATP and translocates the substrate protein into the ClpP chamber by binding and hydrolysis of ATP. At single molecular level, ClpX harnesses cycles of power stroke (dwell and burst) to unfold the substrates, then releases the ADP and Pi. Based on the construction and function of ClpXP, especially the recent progress on how ClpX unfold protein substrates, in this mini-review, a currently proposed single ClpX molecular model system detected by optical tweezers, and its prospective for the elucidation of the mechanism of force generation of ClpX in its power stroke and the subunit interaction with each other, were discussed in detail.

Isolation and Identification of Endophytic Fungi in Kernels of Coix lachrymal-jobi L. Cultivars.

Coix lachrymal-jobi L. var. ma-yuen Stapf of Gramineae are annual or perennial herbs and an important food-medicine homologous plants of high value in nutrition, health protection, and comprehensive utilization. In recent years, the revival of researches on its roles in food and medicinal applications of this underutilized grass for food security and economic empowerment of rural communities has been seen . In this research, Coix kernel endophytic fungi were isolated and identified by fungal colony morphology observation combined with the PCR-amplified fungal internal transcribed spacer (ITS) sequence analyses. All together six isolates to five species of Coix endophytic fungi and two isolates to the genus level were identified from the kernels of six Coix cultivars: Penicillium expansum, Penicillium polonicum, Cladosporium cladosporioides, Alternaria alternata, Aspergillus flavus, and two genera of Aspergillus and Fusarium. Potential benefits and harms analyses showed that Penicillium expansum, Aspergillus oryzae, and Cladosporium cladosporioides can produce a variety of beneficial composite enzymes and have an extensive application in microbial chemistry, food science, and fermentation, whereas Penicillium, Aspergillus flavus, Alternaria alternate, and Fusarium can produce corresponding toxins harmful to plants, animals, and humans. These results not only provided a basis for the targeted prevention of contamination in the tissue culture of Coix kernels by the addition of specific antibiotics, but also enriched the endophytic fungi resource pool of Gramineae crops and suggested new ideas for the improvement, cultivation, post-harvest seeds/kernels storage, and the development of new natural drugs.

Single and joint toxic effects of four antibiotics on some metabolic pathways of zebrafish (Danio rerio) larvae.

In China, antibiotics are commonly used for human and veterinary medicine, and they are present in various environmental media. Thus, the toxic effects of antibiotics on organisms have attracted the attention of society and scientists alike. In this study, zebrafish embryos were used to test the single and joint toxicity of four antibiotics, sulfamonomethoxine (SMM), cefotaxime sodium (CFT), tetracycline (TC), enrofloxacin (ENR), and their combinations, combining the results of experimental and omics techniques. Following exposure to antibiotics for 120 h, the body lengths of zebrafish larvae in all 100 µg/L antibiotic groups were significantly shortened, and the reactive oxygen species (ROS) content in the 100 µg/L Mix group was significantly increased. Transcriptome sequencing (RNA-seq) showed that the mRNA level of numerous genes was significantly changed in the five antibiotic treatment groups. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the differentially expressed genes revealed a significant enrichment of the steroid biosynthesis and other metabolism pathways. Hub gene analysis highlighted dhcr24, acat1, aldh1a2, aldh8a1, suclg2, hadh, and hsdl2 as the key genes, and hub gene expression changes because of the antibiotic treatment suggested that the metabolic system of the zebrafish larvae was severely disrupted by the interaction with other genes. In conclusion, single or joint exposure to different antibiotics at environmental concentrations affected the early development and metabolic system of zebrafish larvae, and our results provide fundamental evidence for future studies of antibiotic toxicity in aquatic organisms.

Transcriptomic analysis of bisphenol AF on early growth and development of zebrafish (Danio rerio) larvae.

Bisphenol AF (BPAF), an alternative to bisphenol A, is widely detected in aquatic environments. Owing to health concerns, the toxic effects of BPAF on organisms are drawing attention. The present study aims to evaluate the toxicity of BPAF, combining the results of omics techniques and experiment. Employing transcriptome sequencing (RNA-seq), we obtained 391, 648, 512, and 545 differentially expressed genes (DEGs) in 0.1, 1, 10, and 100 µg/L BPAF-exposed zebrafish larvae, respectively. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment revealed the early development, stimulus-response, and MAPK signaling pathway were significantly affected by BPAF. In addition, five hub genes (fgf3, fgf4, map2k1, myca, and casp3b) were highlighted as the key genes in MAPK signaling pathway using the protein-protein interaction network. Therefore, the RNA-seq results showed that early development and stimulus-response were the main processes affected by BPAF, which was consistent with our morphological and pathological results. The hatching rate of zebrafish embryos in 1 and 10 µg/L BPAF groups was significantly inhibited, and the oxidative stress indexes, including the level of total antioxidant capacity (T-AOC), superoxide dismutase (SOD), and lipid peroxidation (LPO), were significantly increased by the 100 µg/L BPAF treatment. Moreover, the activity of alkaline phosphatase (AKP) was significantly decreased in all BPAF exposure groups. In conclusion, exposure to BPAF at environmental relevant concentrations affected the early development and immune system of zebrafish larvae by modulating MAPK signaling pathway, and our results provide solid evidence for the future studies on the toxicity of bisphenols.

Z-Ligustilide Exerted Hormetic Effect on Growth and Detoxification Enzymes of Spodoptera litura Larvae.

Plants have evolved a variety of phytochemicals to defense insect feeding, whereas insects have also evolved diverse detoxification enzymes, which are adaptively induced as a prosurvival mechanism. Herein, Z-ligustilide in Ligusticum chuanxiong Hort. was found to exhibit a similar trend in the accumulation from December to May as the occurrence of Spodoptera litura (Fabricius) larvae. Importantly, S. litura larvae feeding enhanced Z-ligustilide level in the stem and leaf (p < 0.01). Moreover, Z-ligustilide ranging from 1 to 5 mg·g-1 exhibited remarkable larvicidal activity, antifeedant activity, and growth inhibition against S. litura larvae. The LC50 values of larvicidal activity for phthalides in L. chuanxiong were compared as follows: Z-ligustilide > levistilide A > senkyunolide A > 3-butylidenephthalide > senkyunolide I, implicating the critical role of conjugated structure. Notably, there was a biphasic dose response for glutathione S-transferase (GST), cytochrome P450 (CYP) 450, Acetylcholinesterase (AChE), and Carboxylesterase (CarE) activities and GSTs1, cytochrome P450 (CYP) 4S9, and CYP4M14 mRNA expression. Particularly, low dose (0.1 mg·g-1) of Z-ligustilide conferred the resistance of S. litura larvae against chlorpyrifos (p < 0.05). Together, our data suggest that Z-ligustilide may function in a hormetic way in the chemical defense of L. chuanxiong against S. litura larvae.

Photosynthetic and physiological analysis of the rice high-chlorophyll mutant (Gc).

Chlorophyll (Chl) molecules are essential for harvesting light energy in photosynthesis. A rice high-chlorophyll mutant (Gc) with significantly increased Chl b was identified previously in Zhenshan 97B (Oryza sativa indica). However, the mechanism underlying this higher Chl b content and its effects on photosynthetic efficiency are still unclear. Immunoblot and blue native polyacrylamide gel electrophoresis (BN-PAGE) with a second dimension electrophoresis followed by the matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF) analysis showed that a few core proteins of photosystem I (PSI) and photosystem II (PSII), and light-harvesting complex II (LHCII) proteins were overexpressed in the mutant plants. Remarkable differences in chloroplast ultrastructure were observed between the wild-type and mutant plants, with the latter having more highly stacked and larger grana. Chl florescence analysis demonstrated that Gc had markedly increased quantum efficiency of photosystem II (ΦPSII), photochemical quenching (qP), non-photochemical quenching (qN) and electron transport rate (ETR). This morphological and physiological adaptation might confer a higher photosynthetic capacity in Gc than the wild-type.