Gelling Behavior of Plant Proteins and Polysaccharides in Food Systems

Biological macromolecules, such as proteins and polysaccharides, are widely used in food systems because their interactions impart a desirable texture to food products. Plant proteins interact with food components via protein-protein and protein-polysaccharide associations, and the formation of a matrix, which can entrap other food components such as water, lipids and flavors. These networks provide structural integrity to food products and can serve as important functional ingredients in processed foods. Intermolecular interactions of typical polysaccharides result either in simple associations or in the form of a double or triple helix. The linear double helical segments may then interact to form a super junction and a three-dimensional gel network. The formation of these structural networks takes place during processing and involves the transformation from a liquid or viscous sol into a solid material with elastic properties. Interests in the behavior of mixed gels center on the prospects of enhanced flexibility in their mechanical and structural properties compared to those of pure gels. Findings on molecular interactions between plant proteins （e.g., soy, canola and pea proteins） and polysaccharides （e.g., guar gum, carrageenan, and pectin） allow for the modification of physical and textural characteristics of mixed biopolymers to meet desired functional property.

Research Progress on Plant Anti-Freeze Proteins

Plant antifreeze proteins(AFPs)are special proteins that can protect plant cells from ice crystal damage in low-temperature environments,and they play a crucial role in the process of plants adapting to cold environ-ments.Proteins with these characteristics have been found infish living in cold regions,as well as many plants and insects.Although research on plant AFPs started relatively late,their application prospects are broad,leading to the attention of many researchers to the isolation,cloning,and genetic improvement of plant AFP genes.Studies have found that the distribution of AFPs in different species seems to be the result of independent evolu-tionary events.Unlike the AFPs found infish and insects,plant AFPs have multiple hydrophilic ice-binding domains,and their recrystallization inhibition activity is about 10–100 times that offish and insect AFPs.Although different plant AFPs have the characteristics of low TH and high RI,their DNA and amino acid sequences are completely different,with small homology.With in-depth research and analysis of the character-istics and mechanisms of plant AFPs,not only has our understanding of plant antifreeze mechanisms been enriched,but it can also be used to improve crop varieties and enhance their freezing tolerance,yield,and quality through genetic engineering.In addition,the study of plant AFPs also contributes to our understanding of freezing resistance mechanisms in other organisms and provides new research directions for thefield of biotech-nology.Therefore,based on the analysis of relevant literature,this article will delve into the concepts,character-istics,research methods,and mechanisms of plant AFPs,summarize the latest research progress and application prospects of AFPs in plant,and provide prospects for the future development of AFP gene research.

Adulteration detection of plant protein beverages by UPLC-MS/MS based on signature peptide of allergen

Plant protein beverage adulteration occurs frequently,which may cause health problems for consumers due to the hidden allergens.Hence,a novel method was developed for authentication by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).Almond,peanut,walnut and soybean were hydrolyzed,followed by separation by NanoLC-Triple TOF MS.The obtained fingerprints were identified by ProteinPilotTM combined with Uniprot,and 16 signature peptides were selected.Afterwards,plant protein beverages treated by trypsin hydrolysis were analyzed with UPLC-MS/MS.This method showed a good linear relationship with R2>0.99403.The limit of quantification(LOQ)were 0.015,0.01,0.5 and 0.05 g/L for almond,peanut,walnut and soybean,respectively.Mean recoveries ranged from 84.77%to 110.44%with RSDs<15%.The developed method was successfully applied to the adulteration detection of 31 plant protein beverages to reveal adulteration and false labeling.Conclusively,this method could provide technical support for authentication of plant protein beverages to protect the rights and health of consumers.

The meat of the matter: plant-based meat analogue versus traditional meat product nutritional quality

Plant-based meat analogues(PBMA)are formulated to mimic the sensory characteristics of traditional meat products(TMP)using vegetarian ingredients and are increasingly popular with the trend towards plantbased diets,even among meat-eating consumers.This narrative review compared the nutritional quality of PBMA and TMP to broaden the discussion on the suitability of simulated meat products to substitute TMP,while exploring other attributes for their comparison.The complexity of PBMA and TMP means they are not entirely interchangeable.Importantly,PBMA health effects extend beyond their basic nutrient content and quality.Postprandial metabolism and satiety,for example,can be affected by product formulation,ingredient interactions,and thermomechanical processing involved in producing organoleptically acceptable PBMA.There are specific opportunities for low-sodium PBMA products to contribute to a healthy shift toward plantbased diets.In conclusion,PBMA cannot be assumed to be healthier than TMP.They should be designed with comprehensive consideration of composition and processing to ensure they support consumers who are adopting plant-based diets in realizing the theoretical health benefits.

Classification and Nomenclature of Plant Metallothionein-like Proteins Based on Their Cysteine Arrangement Patterns

随着植物基因组研究的进展 ,在基因文库和蛋白文库登录的植物类金属硫蛋白基因已超过 5 0个 ,接近金属硫蛋白总数的 1/ 3,而且有不断上升的趋势。鉴于目前植物类金属硫蛋白命名与分类随意性太大 ,很有必要建立一个统一合理的命名与分类法。对植物类金属硫蛋白一级结构进行详细分析后 ,发现该蛋白两端富含半胱氨酸的区域内半胱氨酸的排列方式颇具规律性 ,进而提出了以半胱氨酸排列方式为基础的分类及命名法 ,并阐述了采用这种方法的理由及其可行性。

Higher Plant Proteins of Cyanobacterial Origin:Are They or Are They Not Preferentially Targeted to Chloroplasts?

Dear Editor, What does the evolutionary origin of a plant protein tell about its subcellular localization？ Naively thinking, one would assume that plant proteins that were originally encoded in the endosymbiont genome are targeted to the chloroplast. However, published data seem to support only a loose link between evolutionary origin and subcel- lular localization. About half of the Arabidopsis proteins with a detectable cyanobacterial ortholog are targeted to subcellular compartments other than the chloroplast （Martin et al., 2002）. H

Plant Antifreeze Proteins and Their Expression Regulatory Mechanism

Low temperature is one of the major limiting environmental factors which constitutes the growth, development, productivity and distribution of plants. Over the past several years, the proteins and genes associated with freezing resistance of plants have been widely studied. The recent progress of domestic and foreign research on plant antifreeze proteins and the identifica- tion and characterization of plant antifreeze protein genes, especially on expression regulatory mechanism of plant antifreeze proteins are reviewed in this paper. Finally, some unsolved problems and the trend of research in physiological functions and gene expression regulatory mechanism of plant antifreeze proteins are discussed.

Effect of Plant AntifreezeProteins on Porcine Embryo to Cryopreservaton

This study was to research the cryopreservation effect of plant antifieeze proteins(AFPs) on day 7 porcine expanded and hatched blastocysts that were frozen in 1.5 M glycerol by conventional slow freezing method. The developmental rates of porcine embryos frozen with 0.5 mg/ml AFPs in freeze medium and cultured for 12 and 24 hours in vitro are 25% and 0%respectively. With the concentration of AFPs increased to 5 mg/ml, the corresponding values became 80% and 40%. The hatched rates for porcine embryos frozen with 0.5 mg/ml and 5 mg/ml of AFPs and cultured for 24 hours in vitro are 0% and 20% respectively..The developmental and hatched rate of the contnrol are all 0%(0/4). These results indicate that the survival rates of porcine expanded and hatched blastocyst can be improved by supplementing freeze medium with plant AFPs.

Estimation of Digestible Energy Values of Plant Protein Supplement in Pig

[Objective] The aim of the study was to establish the effective and accurate formulas for estimating the digestible energy （DE） values of plant protein supplement in pig. [Method] By difference method with different amount of alternative feeds （20% -50%）, two4 x4 Latin- square-designed trials were taken on eight castrated male pigs [ Yorkshire x Landrace x Neijiang pig, initial body-weight： （46 ±2） kg ] to deter- mine the apparent digestible energy （ADE） of the eight kinds of plant protein supplement commonly used in China, that is, corn gluten meal （sol.）, soybean meal （ sol. ）, fababean, pea, rapeseed meal （ sol. ）, sesame meal （ sol. ）, rapeseed meal （ exp. ） and cotton seed meal （sol.）. [Resultl （1） Fiber was the most important factor to estimate the ADE of plant protein supplement in pigs, and ADF was the best one. （2） The most effective equations were as below： （ 1 ） OE （kJ/kg DM） = 14 741.86 - 185.01ADF＋54.01SCHO＋22.45CP （ R =0.988,RSD= 67.9,P〈0.01 ） ; （2） DE （kJ/kg DM） =22 223.26 -209.58ADF＋26.79SCHO-1.09GE （ Ff =0.989,RSD=66.9, P〈0.01 ） . [Conclusion] The accurate, practical and specific regression equations were established for DE prediction of plant protein supplement in pig.

Emerging technological developments to address pest resistance in Bt cotton

Cotton plays a crucial role in shaping Indian economy and rural livelihoods.The cotton crop is prone to numerous insect pests,necessitating insecticidal application,which increases production costs.The advent of the expression of Bacillus thuringiensis(Bt)insecticidal protein in cotton has significantly reduced the burden of pest without compromising environmental or human health.After the introduction of transgenic cotton,the cultivated area expanded to 22 million hectares,with a 64% increase in adoption by farmers worldwide.Currently,Bt cotton accounts for 93% of the cultivated cotton area in India.However,extensive use of Bt cotton has accelerated resistance development in pests like the pink bollworm.Furthermore,the overreliance on Bt cotton has reduced the use of broad-spectrum pesticides,favouring the emergence of secondary pests with significant challenges.This emphasizes the urgent necessity for developing novel pest management strategies.The high-dose and refuge strategy was initially effective for managing pest resistance in Bt cotton,but its implementation in India faced challenges due to misunderstandings about the use of non-Bt refuge crops.Although gene pyramiding was introduced as a solution,combining mono toxin also led to instances of cross-resistance.Therefore,there is a need for further exploration of biotechnological approaches to manage insect resistance in Bt cotton.Advanced biotechnological strategies,such as sterile insect release,RNA interference(RNAi)-mediated gene silencing,stacking Bt with RNAi,and genome editing using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR-Cas),offer promising tools for identifying and managing resistance genes in insects.Additionally,CRISPR-mediated gene drives and the development of novel biopesticides present potential avenues for effective pest management in cotton cultivation.These innovative approaches could significantly enhance the sustainability and efficacy of pest resistance management in Bt cotton.

Modeling the Cysteine Rich Domain of Plant Metallothionein-like Protein

With the progress of plant genome research, more than 50 plant metallothionein_like (MT_L) genes have been found, but only several MT_L proteins have been detected and no experimental structural information for MT_L proteins has been reported so far. Since detailed knowledge of the protein tertiary structure is required to understand its biological function, a method is needed to determine the structure of these proteins. In this study, the structural data of known mammal MT was used to determine the interatomic distance constraints of the CXC and CXXC motifs and the metal_sulfur chelating cluster. Then several possible MT conformations were predicted using a distance geometry algorithm. The statistical analysis was used to select those with much lower target function values and lower conformation energies as the predicted tertiary structural models of the cysteine_rich (CR) domains of these proteins. A suitable prediction method for modeling the CR domain of the plant MT_L protein was constructed. The accurately predicted result for the known structure of an MT protein from blue crab suggests that this method is practicable. The tertiary structures of CR domains of rape MT_L protein LSC54 was then modeled with this method.

The potential of legume-derived proteins in the food industry

Legume-derived proteins present an opportunity to replace existing animal-source protein in various applications. Those proteins are abundant, relatively lowcost, sustainable, not highly allergenic, and widely acceptable by consumers. In this paper, it was found that legume-derived protein’s technofunctional properties(e.g. gelation, emulsification and foaming) are being investigated, in order to assess their ability to substitute for animal-derived proteins in food systems and applications. This paper reviewed the functional attributes of legume-derived proteins, their possible applications in food systems, and their potential in the food industry. The techno-functional properties of the proteins vary among different legumes, and while some proteins properties are sufficient for industrial uses, the other may need to be modified. It was concluded that legume-derived proteins could replace some existing animal-derived protein based food systems.

Cloning and Expression Patterns of a Metallothionein-like GenehtMT2 of Helianthus tuberosus

A novel cDNA sequencehtMT2, which encodes a type 2 metallothionein_like protein, was isolated from Helianthus tuberosus L. tuber cDNA library. The whole sequence is 509 bp, including an open reading frame (ORF) of 240 bp, a 5′ UTR of 62 bp and a 3′ UTR of 207 bp. Two genomic sequences covering the coding region ofhtMT2were cloned by PCR reaction. Sequence analysis revealed that the genomic sequences htMTG_1 of 986 bp and htMTG_2 of 982 bp were both composed of three exons and two introns. The deduced protein consisted of 79 amino acid residues with a predicted molecular weight of 7.8 ku (kD). Amino_terminal and carboxy_terminal domains contained 8 and 7 cysteine residues respectively, separated by a central cysteine free spacer. Sequence alignment revealed that the predicted protein ofhtMT2 was homologous to type 2 metallothioneins (MTs) of plants. Southern blotting analysis indicated that htMT2was encoded by a small multi_gene family in H. tuberosus genome. Northern blotting analysis showed that htMT2 transcripts were detected in stems, leaves and leafstalks, but no transcripts were detected in roots. The expression level in stems was the highest among the above tissues. Transcripts in stems were significantly reduced by Cu 2+ treatment. Judging from the homologies between the deduced HtMT2 and other type 2 plant metallothioneins as well as responses to metal ions, we believe thatwere cloned by PCR reaction. Sequence analysis revealed that the genomic sequences htMTG_1 of 986 bp and htMTG_2 of 982 bp were both composed of three exons and two introns. The deduced protein consisted of 79 amino acid residues with a predicted molecular weight of 7.8 ku (kD). Amino_terminal and carboxy_terminal domains contained 8 and 7 cysteine residues respectively, separated by a central cysteine free spacer. Sequence alignment revealed that the predicted protein ofhtMT2 was homologous to type 2 metallothioneins (MTs) of plants. Southern blotting analysis indicated that htMT2was encoded by a small multi_gene family in H. tuberosus genome. Northern blotting analysis showed that htMT2 transcripts were detected in stems, leaves and leafstalks, but no transcripts were detected in roots. The expression level in stems was the highest among the above tissues. Transcripts in stems were significantly reduced by Cu 2+ treatment. Judging from the homologies between the deduced HtMT2 and other type 2 plant metallothioneins as well as responses to metal ions, we believe that[ShtMT2 encodes a new type 2 metallothionein.

Pathogenesis of coeliac disease:implications for treatment

INTRODUCTIONCoeliac disease(CD)is an enteropathy ,characterised by villous atrohy ,which occurs in genetically susceptible individuals .It affects mainly the proximal small intestine,and is caused by an intolerance to cereal storage proteins found in wheat ,barley and rye .

Partial and Total Replacement of Jojoba Meal （Simmondsia chinensis） Instead of Fishmeal in Nile Tilapia （Oreochromis niloticus） Fingerlings Diets

The present study was carried out to investigate the total and partial replacement of Jojoba meal （JM）, Simmondsia chinensis, with fishmeal （FM） at different levels （0%, 25%, 50%, 75%, and 100%） on growth performance, feed utilization, and carcass composition of Nile tilapia, Oreochromis niloticus fingerlings. Fingerlings with an average weight of 1.65 ± 0.01 g/fish stocked at a rate of 10 fish/aquarium for 84 days experimental period. All experimental diets were isocaloric （437.69 kcal/100 g Dry Matter, DM） and isonitrogenous （30.5% crude protein） and supplemented with 0.5% L-Methionine and 0.5% L-Lysine Hcl. Results showed that there were a significant differences （P 〈 0.05） in final body weights, average daily gain （g fish^-1 dayl）, specific growth rate （SGR, % day^-1）, feed conversion ratio （FCR）, protein efficiency ratio （PER）, protein productive value （PPV%） and energy retention （ER%） among the tested groups. Nile tilapia fingerlings fed on the diet containing 25% S. chinensis protein exhibited comparable growth performance to those fed FM protein based diet. Carcass composition of crude protein decreased significantly （P 〈_ 0.05） with increasing JM replacement level above 50%, while replacement 25% JM does not affect DM and Crude Protein of fish flesh. On the other hand, increasing JM replacement level up to 75% increased Ether Extract % significantly （P 〈 0.05） and increasing JM up to 100% increased ash content significantly （P ≤ 0.05） while energy content （EC） decreased. It could be concluded the replacement of 25% Jojoba meal instead of fishmeal in Nile tilapia fingerlings diets is possible without any adverse effects on its growth performance or feed utilization.

Proline with or without Hydroxyproline Influences Collagen Concentration and Regulates Prolyl 4-Hydroxylase α(I) Gene Expression in Juvenile Turbot(Scophthalmus maximus L.)

This study was conducted to investigate the effect of dietary proline(Pro), and Pro and hydroxyproline(Hyp) in combination on the growth performance, total Hyp and collagen concentrations of tissues, and prolyl 4-hydroxylase α(I)(P4H α(I)) gene expression in juvenile turbot feeding high plant protein diets. A diet containing 50% crude protein and 12% crude lipid was formulated as the basal and control, on which other two protein and lipid contents identical experimental diets were formulated by supplementing the basal with either 0.75% Pro(Pro-0.75) or 0.75% Pro and 0.75% Hyp(Pro+Hyp). Four groups of fish in indoor seawater recirculating systems, 35 individuals each, were fed twice a day to apparent satiation for 10 weeks. The results showed that dietary Pro and Hyp supplementation had no significant effect on growth performance and feed utilization of juvenile turbot(P > 0.05). Total Hyp and collagen concentrations in muscle were significantly increased when dietary Pro and Hyp increased(P < 0.05), and fish fed diet Pro+Hyp showed significantly higher free Hyp content in plasma than those fed other diets(P < 0.05). The expression of P4 H α(I) gene in liver and muscle was significantly up regulated in fish fed diet Pro-0.75 in comparison with control(P < 0.05); however the gene was significantly down regulated in fish fed diet Pro+Hyp in muscle in comparison with fish fed diet Pro-0.75(P < 0.05). It can be concluded that supplement of crystal L-Pro and L-Hyp to high plant protein diets did not show positive effects on growth performance of juvenile turbot, but enhanced total collagen concentrations in muscle.

Replacing fishmeal with soybean meal affects survival, antioxidant capacity, intestinal microbiota, and mRNA expression of TOR and S6K1 in Macrobrachium rosenbergii

Using alternative plant-derived dietary protein to replace fishmeal,combined with practical evaluation indexes,is a recent focus for aquaculture practices.An 8-week feeding experiment with giant freshwater prawn Macrobrachium rosenbergii post-larvae was conducted to determine the eff ects of replacing fi shmeal(FM)with soybean meal in the feed,in terms of growth performance,antioxidant capacity,intestinal microbiota,and mRNA expression of target of rapamycin(TOR)and ribosomal protein S6 kinase B1(S6K1).Four isonitrogenous diets with isocaloric value were prepared to contain 100%,75%,50%,or 25%FM as the protein source(dietary treatments FM100,FM75,FM50,and FM25,respectively).Each diet was fed to post-larval prawns(mean weight 0.045±0.002 g)twice a day in four replicates.No signifi cant diff erence in weight gain was observed among all groups,but the survival rate of prawns fed the FM50 and FM25 diets was signifi cantly lower than that of prawns fed the FM diet.The mRNA expression of both TOR and S6K1 were the lowest in hepatopancreas of prawns fed the FM25 diet.Superoxide dismutase activity of prawns fed the FM25 diet was significantly lower than that of prawns fed FM50.In contrast,the malondialdehyde content was signifi cantly higher in prawns fed FM25 as compared with those fed FM75.The proportion of fishmeal in the diet did not affect the composition of core(phylum-level)intestinal microbiota,but greater fishmeal replacement with soybean meal had a potential risk to increase the relative abundance of opportunistic pathogens in the gut when considered at the genus level.These results suggest that fishmeal replacement with soybean meal should not exceed 50%in a diet for post-larval M.rosenbergii.

Plant glycosylphosphatidylinositol anchored proteins at the plasma membrane-cell wall nexus

Approximately 1% of plant proteins are predicted to be post-translationally modified with a glycosylphospha- tidylinositol （GPI） anchor that tethers the polypeptide to the outer leaflet of the plasma membrane. Whereas the synthesis and structure of GPI anchors is largely conserved across eukaryotes, the repertoire of functional domains present in the GPl-anchored proteome has diverged substantially. In plants, this includes a large fraction of the GPl-anchored proteome being further modified with plant-specific arabinogalactan （AG） O-glycans. The impor- tance of the GPl-anchored proteome to plant development is underscored by the fact that GPI biosynthetic null mutants exhibit embryo lethality. Mutations in genes encoding specific GPl-anchored proteins （GAPs） further supports their contribution to diverse biological processes, occurring at the interface of the plasma membrane and cell wall, including signaling, cell wall metabolism, cell wall polymer cross-linking, and plasmodesmatal transport. Here, we review the literature concerning plant GPl-anchored proteins, in the context of their potential to act as molecular hubs that mediate interactions between the plasma membrane and the cell wall, and their potential to transduce the signal into the protoplast and, thereby, activate signal transduction pathways.

Effects of temperature and particle size on the thermophysical properties of six plant-origin protein supplements

Reasonable design of the parameters of thermal processing such as conditioning and cooling according to formula changes of pelleted feeds has always been a serious challenge for Chinese feed mills and feed equipment manufacturers. Studying the thermophysical properties of different protein feeds under different temperatures and particle sizes will facilitate the equipment design, parameter optimization, and simulation for the thermal processing of pelleted feeds. In this study, the specific heat (Cp), thermal conductivity (kb), and thermal diffusivity (α) of six plant protein supplements with three particle sizes were determined over a temperature range of 25℃-100℃. The differences in Cp, kb, and α among different feedstuffs and particle sizes were analyzed and the influences of temperature and particle size on these properties were evaluated. Results showed that the Cp, kb, and α of all the feedstuffs increased with increasing temperature and varied from 1.622 to 2.417 kJ/(kg∙℃), 0.080 to 0.362 W/(m∙℃), 6.379×10^(-8) to 21.984×10^(-8) m^(2)/s, respectively. To rise to the same temperature, the distiller’s dried grain with solubles (DDGS) needed to absorb 3% more heat than that required for soybean meal (SBM), while the rest four feedstuffs just needed to absorb 93%-98% heat for SBM. Particle size had no significant effect on Cp for all the feedstuffs (p>0.05). However, descending trends in kb and α were observed with increasing particle size for a certain feedstuff at the same bulk density. In addition, regression equations with only statistically significant terms were developed to describe Cp, kb, and α as a function of temperature and particle size for six feedstuffs. The results can provide basic theory and data for the optimization of thermal processing parameters required for the plant-protein ingredient change in compound feed formulations.

Gossypol is the main limiting factor in the application of cottonseed meal in grass carp feed production:Involvement of growth,intestinal physical and immune barrier,and intestinal microbiota

The objective of this study is to investigate the effects of free gossypol in cottonseed meal(CM)on the growth and intestinal health of grass carp,as well as to determine whether free gossypol is the primary factor restricting highlevel inclusion of cottonseed meal.This study was designed with five groups:the control group(32%soybean concentrate protein),the CM group(44%CM),the cottonseed protein concentrate(CPC)group(33%CPC),and the CPC and control groups with 400 and 600 mg/kg gossypol added,designated as CON,CM,CPC,CPC+GP,and CON+GP,respectively.The experiment lasted eight weeks,and each group had three replicates.This experiment employed one-way ANOVA and Duncan's multiple comparisons of the means.The findings revealed that in comparison to the CON group,the growth of fish in the CM,CPC+GP,and CON+GP groups significantly decreased.Intestinal inflammation damage was observed.This was indicated by a significant upregulation of proinflammatory factors,including tnf-α,nf-κb,il-6,il-8,il-12β,and il-1β,along with significant downregulation of anti-inflammatory factors,including tgf-β1,il-15,and il-10.Additionally,significant downregulation of antioxidant enzyme-related genes,including nrf2,cat,CuZnsod,gpx4,and gpx1,was observed and the intestinal physical barrier function was compromised.In addition,the intestinal microbiota composition was affected,with a significant reduction in Bacillus and Cetobacterium abundances and a remarkable increase in the abundance of Aeromonas,resulting in dysregulation of the intestinal microbiota function.However,in comparison to the CM group,the growth rate of fish in the CPC group exhibited a marked increase;nonetheless,it persisted at a level lower than that observed in the CON group and the degree of intestinal damage was significantly improved.Additionally,the intestinal microbiota structure was found to be similar to that of the CON group.In conclusion,excessive CM negatively affects grass carp growth and intestinal health,whereas replacing CM with CPC mitigates these effects.Adding gossypol equivalent to CM group in CON and CPC diets reduces growth performance and impairs intestinal function,indicating gossypol as a primary limiting factor for high CM inclusion in diets.