Parenteral Vaccination with a Live Brucella melitensis Mutant Protects against Wild-Type B. melitensis 16M Challenge.

Susceptibility to brucellosis remains prevalent, even in herds vaccinated with conventional vaccines. Efforts are underway to develop an improved brucellosis vaccine, and possibly a universal vaccine, given that Brucella species are highly homologous. To this end, two B. melitensis mutants were developed, znBM-lacZ (znBMZ) and znBM-mCherry (znBM-mC), and were tested for their ability to confer systemic immunity against virulent B. melitensis challenge. To assess the extent of their attenuation, bone-marrow-derived macrophages and human TF-1 myeloid cells were infected with both mutants, and the inability to replicate within these cells was noted. Mice infected with varying doses of znBM-mC cleared the brucellae within 6-10 weeks. To test for efficacy against systemic disease, groups of mice were vaccinated once by the intraperitoneal route with either znBMZ or B. abortus S19 vaccine. Relative to the PBS-dosed mice, znBMZ vaccination greatly reduced splenic brucellae colonization by ~25,000-fold compared to 700-fold for S19-vaccinated mice. Not surprisingly, both znBMZ and S19 strains induced IFN-γ+ CD4+ T cells, yet only znBMZ induced IFN-γ+ CD8+ T cells. While both strains induced CD4+ effector memory T cells (Tems), only znBMZ induced CD8+ Tems. Thus, these results show that the described znBM mutants are safe, able to elicit CD4+ and CD8+ T cell immunity without a boost, and highly effective, rendering them promising vaccine candidates for livestock.

A Single Nasal Dose Vaccination with a Brucella abortus Mutant Potently Protects against Pulmonary Infection.

The Brucella abortus double-mutant (ΔznuA ΔnorD Brucella abortus-lacZ [znBAZ]) was assessed for its protective efficacy after vaccination with a single nasal dose. Superior protection was achieved in znBAZ-vaccinated mice against pulmonary, wild-type B. abortus 2308 challenge when compared with conventional livestock Brucella abortus vaccines, the smooth S19 (smooth B. abortus strain 19 vaccine) and rough RB51 (rough mutant vaccine strain of B. abortus) strains. Nasal znBAZ vaccination reduced splenic and lung colonization by wild-type brucellae by >3-4 logs. In contrast, S19 reduced lung colonization by only 32-fold, and RB51 failed to reduce colonization. One profound attribute of znBAZ vaccination was the >3-fold increase in pulmonary CD8+ T cells when compared with other vaccinated groups. S19 vaccination increased only CD4+ T cells. All vaccines induced IFN-γ and TNF-α production by CD4+ T cells, but only znBAZ vaccination enhanced the recruitment of polyfunctional CD8+ T cells, by >100-fold. IL-17 by both CD4+ and CD8+ T cells was also induced by subsequent znBAZ vaccination. These results demonstrate that, in addition to achieving protective immunity by CD4+ T cells, CD8+ T cells, specifically resident memory T cells, also confer protection against brucellosis. The protection obtained by znBAZ vaccination was attributed to IFN-γ-producing CD8+ T cells, because depletion of CD8+ T cells throughout vaccination and challenge phases abrogated protection. The stimulation of only CD4+ T cells by RB51- and S19-vaccinated mice proved insufficient in protecting against pulmonary B. abortus 2308 challenge. Thus, nasal znBAZ vaccination offers an alternative means to elicit protection against brucellosis.

RAP2.6 enhanced salt stress tolerance by reducing Na+ accumulation and stabilizing the electron transport in Arabidopsis thaliana.

The transcription factors of the AP2/ERF family are involved in plant growth and development and responses to biotic and abiotic stresses. Here, we found RAP2.6, a transcription factor which belongs to the ERF subfamily, was responsive to salt stress in Arabidopsis. Under salt stress conditions, rap2.6 mutant seedlings were the sensitivity deficiency to salt stress which was reflected in higher germination rate and longer root length compared to the wild type. Also, the expressions of salt-related gene including SOS1, SOS2, SOS3, NHX1, NHX3, NHX5 and HKT1 in rap2.6 mutant seedlings were lower than the wild type under salt stress. rap2.6 mutant adult lacked salt stress tolerance based on the results of the phenotype, survival rates and ion leakage. Compared to wild type, rap2.6 mutant adult accumulated more Na+ in leaves and roots while the salt-related gene expressions were lower. In addition, the photosynthetic electron transport and PSII energy distribution in rap2.6 mutant plant leaves had been more seriously affected under salt stress conditions compared to the wild type. In summary, this study identified essential roles of RAP2.6 in regulating salt stress tolerance in Arabidopsis.

StICE1 enhances plant cold tolerance by directly upregulating StLTI6A expression.

KEY MESSAGE: Under cold conditions, StICE1 enhances plant cold tolerance by upregulating StLTI6A expression to maintain the cell membrane stability. Cold stress affects potato plants growth and development, crop productivity and quality. The ICE-CBF-COR regulatory cascade is the well-known pathway in response to cold stress in plants. ICE1, as a MYC-like bHLH transcription factor, can regulate the expressions of CBFs. However, whether ICE1 could regulate other genes still need to be explored. Our results showed that overexpressing ICE1 from potato in Arabidopsis thaliana could enhance plant cold tolerance. Under cold stress, overexpressed StICE1 in plants improved the stability of cell membrane, enhanced scavenging capacity of reactive oxygen species and increased expression levels of CBFs and COR genes. Furthermore, StICE1 could bind to the promoter of StLTI6A gene, which could maintain the stability of the cell membrane, to upregulate StLTI6A expression under cold conditions. Our findings revealed that StICE1 could directly regulate StLTI6A, CBF and COR genes expression to response to cold stress.

Diversity of responses to nitrogen deficiency in distinct wheat genotypes reveals the role of alternative electron flows in photoprotection.

Nitrogen (N) deficiency represents an important limiting factor affecting photosynthetic productivity and the yields of crop plants. Significant reported differences in N use efficiency between the crop species and genotypes provide a good background for the studies of diversity of photosynthetic and photoprotective responses associated with nitrogen deficiency. Using distinct wheat (Triticum aestivum L.) genotypes with previously observed contrasting responses to nitrogen nutrition (cv. Enola and cv. Slomer), we performed advanced analyses of CO2 assimilation, PSII, and PSI photochemistry, also focusing on the heterogeneity of the stress responses in the different leaf levels. Our results confirmed the loss of photosynthetic capacity and enhanced more in lower positions. Non-stomatal limitation of photosynthesis was well reflected by the changes in PSII and PSI photochemistry, including the parameters derived from the fast-fluorescence kinetics. Low photosynthesis in N-deprived leaves, especially in lower positions, was associated with a significant decrease in the activity of alternative electron flows. The exception was the cyclic electron flow around PSI that was enhanced in most of the samples with a low photosynthetic rate. We observed significant genotype-specific responses. An old genotype Slomer with a lower CO2 assimilation rate demonstrated enhanced alternative electron flow and photorespiration capacity. In contrast, a modern, highly productive genotype Enola responded to decreased photosynthesis by a significant increase in nonphotochemical dissipation and cyclic electron flow. Our results illustrate the importance of alternative electron flows for eliminating the excitation pressure at the PSII acceptor side. The decrease in capacity of electron acceptors was balanced by the structural and functional changes of the components of the electron transport chain, leading to a decline of linear electron transport to prevent the overreduction of the PSI acceptor side and related photooxidative damage of photosynthetic structures in leaves exposed to nitrogen deficiency.

StLTO1, a lumen thiol oxidoreductase in Solanum tuberosum L., enhances the cold resistance of potato plants.

Cold stress reduces plant photosynthesis and increases the accumulation of reactive oxygen species (ROS) in plants, thereby dramatically affecting plant growth, crop productivity and quality. Here, we report that lumen thiol oxidoreductase 1 (StLTO1), a vitamin K epoxide reductase (VKOR)-like protein in the thylakoid membrane of Solanum tuberosum L., enhances the cold tolerance of potato plants. Under normal conditions, overexpression of StLTO1 in plants promoted plant growth. In addition, potato plants overexpressing StLTO1 displayed enhanced photosynthetic capacity and increased capacity for scavenging ROS compared to StLTO1 knockdown and wild-type potato plants under cold conditions. Overexpression of StLTO1 in potato plants also improved cold-regulated (COR) gene expression after cold stress. Our results suggest that StLTO1 acts as a positive regulator of cold resistance in potato plants.

Live mucosal vaccination stimulates potent protection via varied CD4+ and CD8+ T cell subsets against wild-type Brucella melitensis 16M challenge.

Re-emerging zoonotic pathogen Brucella spp. continues to impact developing countries and persists in expanding populations of wildlife species in the US, constantly threatening infection of our domestic herds. The development of improved animal and human vaccines remains a priority. In this study, immunity to a novel live attenuated B. melitensis strain, termed znBM-mC, was characterized. An oral prime, intranasal (IN) boost strategy conferred exquisite protection against pulmonary challenge, with wild-type (wt) B. melitensis providing nearly complete protection in the lungs and spleens from brucellae colonization. Vaccination with znBM-mC showed an IFN-γ+ CD8+ T-cell bias in the lungs as opposed to Rev 1-vaccinated mice showing IFN-γ+ CD4+ T-cell inclination. Lung CD4+ and CD8+ effector memory T cells (TEMs) increased over 200-fold; and lung CD4+ and CD8+ resident memory T cells (TRMs) increased more than 250- and 150-fold, respectively. These T cells served as the primary producers of IFN-γ in the lungs, which was essential for vaccine clearance and the predominant cytokine generated pre-and post-challenge with wt B. melitensis 16M; znBM-mC growth could not be arrested in IFN-γ-/- mice. Increases in lung TNF-α and IL-17 were also induced, with IL-17 being mostly derived from CD4+ T cells. Vaccination of CD4-/-, CD8-/-, and B6 mice with znBM-mC conferred full protection in the lungs and spleens post-pulmonary challenge with virulent B. melitensis; vaccination of IL-17-/- mice resulted in the protection of the lungs, but not the spleen. These data demonstrate the efficacy of mucosal vaccine administration for the generation of protective memory T cells against wt B. melitensis.

ScCBF1 plays a stronger role in cold, salt and drought tolerance than StCBF1 in potato (Solanum tuberosum).

Solanum tuberosum (St) and Solanum commersonii (Sc) are two potato varieties with different freezing tolerance. Among them, St is a freezing-sensitive variety and. Sc is a cold-resistant wild potato. CBF/DREB family members mainly function in response to freezing stress. In order to explore the different roles of St C-Repeat Binding Factor1 (StCBF1) and Sc C-Repeat Binding Factor1 (ScCBF1) in potato plants (Solanum tuberosum) under stress conditions, two kinds of potato lines were obtained with ScCBF1 and StCBF1 overexpressing respectively. Phenotypes analysis showed that both overexpressing ScCBF1 and StCBF1 caused smaller leaves, and reduced tuber yield. While the limited phenotypes of StCBF1 lines were more severe than that of ScCBF lines. After freezing treatment, StCBF1 over expression plants grown better than WT plants and worse than ScCBF1 over expression plants. Specifically, compared with wild-type lines, overexpressing ScCBF1 could up-regulate fatty acid desaturase genes, key enzyme of Calvin cycle genes, and antioxidant enzyme genes. Both ScCBF1 and StCBF1 lines showed higher PSII activity, thus maintaining a higher photosynthetic rate under cold stress. In addition, we also found that overexpression ScCBF1 and StCBF1 could also enhance the drought and salt tolerance in potato. In summary, ScCBF1 plays a stronger role in cold, salt, and drought tolerance than StCBF1 in potato (Solanum tuberosum).

StATL2-like could affect growth and cold tolerance of plant by interacting with StCBFs.

KEY MESSAGE: Our results confirmed that StATL2-like could interact with StCBFs and regulate plant growth. Meanwhile, StATL2-like acted as a negative regulator on low-temperature tolerance in plants. As important transcription factors for resisting many kinds of stresses, C-repeat-binding factors (CBF) play a key role in plant low-temperature tolerance by increasing COR genes expressions. Here, we report that StATL2-like, a RING-H2 E3 ubiquitin in Solanum tuberosum L., interacted with StCBF1 and StCBF4, respectively. AtATL2 is a highly homologous gene of StATL2-like in Arabidopsis thaliana. Under normal conditions, atl2 Arabidopsis mutant showed a growth inhibition phenotype while overexpressed StATL2-like in wild type Arabidopsis and atl2 mutant promoted plant growth. Besides, atl2 mutant had better low-temperature tolerance compared with wild type and StATL2-like transgenic lines which demonstrated that StATL2-like acted as a negatively regulator on low-temperature tolerance in plant. Moreover, atl2 mutant improved the scavenging capacity of reactive oxygen species (ROS) and alleviate the damage of photosynthetic system II (PSII) compared with StATL2-like transgenic lines under cold conditions. These results suggested a new component in CBF-dependent pathway to regulate plant growth and response to low-temperature stress in potato plants.

Pre-Acclimation to Elevated Temperature Stabilizes the Activity of Photosystem I in Wheat Plants Exposed to an Episode of Severe Heat Stress.

The importance of high temperature as an environmental factor is growing in proportion to deepening global climate change. The study aims to evaluate the effects of long-term acclimation of plants to elevated temperature on the tolerance of their photosynthetic apparatus to heat stress. Three wheat (Triticum sp. L.) genotypes differing in leaf and photosynthetic traits were analyzed: Thesee, Roter Samtiger Kolbenweizen, and ANK 32A. The pot experiment was established in natural conditions outdoors (non-acclimated variant), from which a part of the plants was placed in foil tunnel with elevated temperature for 14 days (high temperature-acclimated variant). A severe heat stress screening experiment was induced by an exposition of the plans in a growth chamber with artificial light and air temperature up to 45 °C for ~12 h before the measurements. The measurements of leaf photosynthetic CO2 assimilation, stomatal conductance, and rapid kinetics of chlorophyll a fluorescence was performed. The results confirmed that a high temperature drastically reduced the photosynthetic assimilation rate caused by the non-stomatal (biochemical) limitation of photosynthetic processes. On the other hand, the chlorophyll fluorescence indicated only a moderate level of decrease of quantum efficiency of photosystem (PS) II (Fv/Fm parameter), indicating mostly reversible heat stress effects. The heat stress led to a decrease in the number of active PS II reaction centers (RC/ABS) and overall activity o PSII (PIabs) in all genotypes, whereas the PS I (parameter ψREo) was negatively influenced by heat stress in the non-acclimated variant only. Our results showed that the genotypes differ in acclimation capacity to heat stress, and rapid noninvasive techniques may help screen the stress effects and identify more tolerant crop genotypes. The acclimation was demonstrated more at the PS I level, which may be associated with the upregulation of alternative photosynthetic electron transport pathways with clearly protective functions.

The network centered on ICEs play roles in plant cold tolerance, growth and development.

MAIN CONCLUSION: ICEs are key transcription factors in response to cold in plant, they also balance plant growth and stress tolerance. Thus, we systematize the information about ICEs published to date. Low temperature is an important factor affecting plant growth and development. Exposing to cold condition results in a suit of effects on plants including reduction of plant growth and reproduction, and decrease in crop yield and quality. Plants have evolved a series of strategies to deal with cold stress such as reprogramming of the expression of genes and transcription factors. ICEs (Inducer of CBF Expression), as transcription factors regulating CBFs (C-repeat binding factor), play key roles in balancing plant growth and stress tolerance. Studies on ICEs focused on the function of ICEs on cold tolerance, growth and development; post-translational modifications of ICEs and crosstalk between the ICEs and phytohormones. In this review, we focus on systematizing the information published to date. We summarized the main advances of the functions of ICEs on the cold tolerance, growth and development. And we also elaborated the regulation of ICEs protein stability including phosphorylation, ubiquitination and SUMOylation of ICE. Finally, we described the function of ICEs in the crosstalk among different phytohormone signaling pathway and cold stress. This review provides perspectives for ongoing research about cold tolerance, growth and development in plant.

Glycinebetaine mitigates tomato chilling stress by maintaining high-cyclic electron flow rate of photosystem I and stability of photosystem II.

KEY MESSAGE: Glycinebetaine alleviates chilling stress by protecting photosystems I and II in BADH-transgenic and GB-treated tomato plants, which can be an effective strategy for improving crop chilling tolerance. Tomato (Solanum lycopersicum) is one of the most cultivated vegetables in the world, but is highly susceptible to chilling stress and does not naturally accumulate glycinebetaine (GB), one of the most effective stress protectants. The protective mechanisms of GB on photosystem I (PSI) and photosystem II (PSII) against chilling stress, however, remain poorly understood. Here, we address this problem through exogenous GB application and generation of transgenic tomatoes (Moneymaker) with a gene encoding betaine aldehyde dehydrogenase (BADH), which is the key enzyme in the synthesis of GB, from spinach. Our results demonstrated that GB can protect chloroplast ultramicrostructure, alleviate PSII photoinhibition and maintain PSII stability under chilling stress. More importantly, GB increased the electron transfer between QA and QB and the redox potential of QB and maintained a high rate of cyclic electron flow around PSI, contributing to reduced production of reactive oxygen species, thereby mitigating PSI photodamage under chilling stress. Our results highlight the novel roles of GB in enhancing chilling tolerance via the protection of PSI and PSII in BADH transgenic and GB-treated tomato plants under chilling stress. Thus, introducing GB-biosynthetic pathway into tomato and exogenous GB application are effective strategies for improving chilling tolerance.

Activation of mucosal immunity as a novel therapeutic strategy for combating brucellosis.

Brucellosis is a disease of livestock that is commonly asymptomatic until an abortion occurs. Disease in humans results from contact of infected livestock or consumption of contaminated milk or meat. Brucella zoonosis is primarily caused by one of three species that infect livestock, Bacillus abortus in cattle, B. melitensis in goats and sheep, and B. suis in pigs. To aid in disease prophylaxis, livestock vaccines are available, but are only 70% effective; hence, improved vaccines are needed to mitigate disease, particularly in countries where disease remains pervasive. The absence of knowing which proteins confer complete protection limits development of subunit vaccines. Instead, efforts are focused on developing new and improved live, attenuated Brucella vaccines, since these mimic attributes of wild-type Brucella, and stimulate host immune, particularly T helper 1-type responses, required for protection. In considering their development, the new mutants must address Brucella's defense mechanisms normally active to circumvent host immune detection. Vaccination approaches should also consider mode and route of delivery since disease transmission among livestock and humans is believed to occur via the naso-oropharyngeal tissues. By arming the host's mucosal immune defenses with resident memory T cells (TRMs) and by expanding the sources of IFN-γ, brucellae dissemination from the site of infection to systemic tissues can be prevented. In this review, points of discussion focus on understanding the various immune mechanisms involved in disease progression and which immune players are important in fighting disease.

Comparison of photosynthetic activity and heat tolerance between near isogenic lines of wheat with different photosynthetic rates.

Wheat (Triticum aestivum L.) is one of the most important crops in the world, but the yield and quality of wheat are highly susceptible to heat stress, especially during the grain-filling stage. Therefore, it is crucial to select high-yield and high-temperature-resistant varieties for food cultivation. There is a positive correlation between the yield and photosynthetic rate of wheat during the entire grain-filling stage, but few studies have shown that lines with high photosynthetic rates can maintain higher thermotolerance at the same time. In this study, two pairs of wheat near isogenic lines (NILs) with different photosynthetic rates were used for all experiments. Our results indicated that under heat stress, lines with a high photosynthetic rate could maintain the activities of photosystem II (PSII) and key Calvin cycle enzymes in addition to their higher photosynthetic rates. The protein levels of D1 and HSP70 were significantly increased in the highly photosynthetic lines, which contributed to maintaining high photosynthetic rates and ensuring the stability of the Calvin cycle under heat stress. Furthermore, we found that lines with a high photosynthetic rate could maintain high antioxidant enzyme activity to scavenge reactive oxygen species (ROS) and reduce ROS accumulation better than lines with a low photosynthetic rate under high-temperature stress. These findings suggest that lines with high photosynthetic rates can maintain a higher photosynthetic rate despite heat stress and are more thermotolerant than lines with low photosynthetic rates.

Does silicon really matter for the photosynthetic machinery in plants ?

Silicon (Si) is known to alleviate the adverse impact of different abiotic and biotic stresses by different mechanisms including morphological, physiological, and genetic changes. Photosynthesis, one of the most important physiological processes in the plant is sensitive to different stress factors. Several studies have shown that Si ameliorates the stress effects on photosynthesis by protecting photosynthetic machinery and its function. In stressed plants, several photosynthesis-related processes including PSII maximum photochemical quantum yield (Fv/Fm), the yield of photosystem II (φPSII), electron transport rates (ETR), and photochemical quenching (qP) were observed to be regulated when supplemented with Si, which indicates that Si effectively protects the photosynthetic machinery. In addition, studies also suggested that Si is capable enough to maintain the uneven swelling, disintegrated, and missing thylakoid membranes caused during stress. Furthermore, several photosynthesis-related genes were also regulated by Si supplementation. Taking into account the key impact of Si on the evolutionarily conserved process of photosynthesis in plants, this review article is focused on the aspects of silicon and photosynthesis interrelationships during stress and signaling pathways. The assemblages of this discussion shall fulfill the lack of constructive literature related to the influence of Si on one of the most dynamic and important processes of plant life i.e. photosynthesis.

Glycinebetaine mitigates drought stress-induced oxidative damage in pears.

Glycinebetaine (GB) is an osmoprotectant found in plants under environmental stresses that incorporates drought and is associated with drought tolerance in several plants, such as the woody pear. However, how GB improves drought tolerance in pears remains unclear. In the current study, we explored the mechanism by which GB enhances drought tolerance of whole pear plants (Pyrus bretschneideri Redh. cv. Suli) supplied with exogenous GB. The results showed that on the sixth day after withholding water, levels of O2·-, H2O2, malonaldehyde (MDA) and electrolyte leakage in the leaves were substantially increased by 143%, 38%, 134% and 155%, respectively. Exogenous GB treatment was substantially reduced O2·-, H2O2, MDA and electrolyte leakage (38%, 24%, 38% and 36%, respectively) in drought-stressed leaves. Furthermore, exogenous GB induced considerably higher antioxidant enzyme activity in dry-stressed leaves than drought-stressed treatment alone on the sixth day after withholding water, such as superoxide dismutase (SOD) (201%) and peroxidase (POD) (127%). In addition, these GB-induced phenomena led to increased endogenous GB levels in the leaves of the GB 100 + drought and GB 500 + drought treatment groups by 30% and 78%, respectively, compared to drought treatment alone. The findings obtained were confirmed by the results of the disconnected leaf tests, in which GB contributed to a substantial increase in SOD activity and parallel dose- and time-based decreases in MDA levels. These results demonstrate that GB-conferred drought resistance in pears may be due in part to minimizing symptoms of oxidative harm incurred in response to drought by the activities of antioxidants and by reducing the build-up of ROS and lipid peroxidation.

Progressive Genomic Approaches to Explore Drought- and Salt-Induced Oxidative Stress Responses in Plants under Changing Climate.

Drought and salinity are the major environmental abiotic stresses that negatively impact crop development and yield. To improve yields under abiotic stress conditions, drought- and salinity-tolerant crops are key to support world crop production and mitigate the demand of the growing world population. Nevertheless, plant responses to abiotic stresses are highly complex and controlled by networks of genetic and ecological factors that are the main targets of crop breeding programs. Several genomics strategies are employed to improve crop productivity under abiotic stress conditions, but traditional techniques are not sufficient to prevent stress-related losses in productivity. Within the last decade, modern genomics studies have advanced our capabilities of improving crop genetics, especially those traits relevant to abiotic stress management. This review provided updated and comprehensive knowledge concerning all possible combinations of advanced genomics tools and the gene regulatory network of reactive oxygen species homeostasis for the appropriate planning of future breeding programs, which will assist sustainable crop production under salinity and drought conditions.

Glycinebetaine: a versatile protectant to improve rice performance against aluminium stress by regulating aluminium uptake and translocation.

KEY MESSAGE: Glycinebetaine alleviates the detrimental effects of aluminium stress by regulating aluminium uptake and translocation, maintaining PSII activity, and activating the oxidative defence, thereby maintaining the growth and development of rice. Aluminium (Al) toxicity is one of the primary growth-limiting factors that limits plant growth and crop productivity in acidic soils. Rice (Oryza sativa L.) plants are susceptible to Al stress and do not naturally accumulate glycinebetaine (GB), one of the most effective protectants. Therefore, the objective of this study was to investigate whether exogenous GB can ameliorate the detrimental effects of Al stress on rice plants. Our results showed that the growth, development and biomass of rice were clearly inhibited under Al stress. However, exogenous GB application increased rice shoot growth and photosynthetic pigments contents, maintained photosystem II (PSII) activity, and activated the antioxidant defence system under Al stress. More importantly, GB may mediate the expression of Al uptake- and translocation-related genes, including OsALS1, OsNrat1, OsSTAR1 and OsSTAR2, and the galacturonic acid contents in rice roots under Al stress. Therefore, our findings highlight exogenous GB application is a valid approach to effectively combat Al toxicity by regulating physiological and biochemical processes in crops.