CBL10 was shown to be a key gene for salt tolerance in Arabidopsis thaliana. In this study, we evaluated the role of CBL10 in the tobacco salt tolerance response by characterizing the gene editing-induced loss-of-function knockout mutants of the two NtCBL10 homeologous genes NtCBL10A and NtCBL10B. The importance of NtCBL10 for the response to salinity was evidenced by the salt supersensitivity of the Nt-cbl10a10b double mutants, with fast-developing chlorosis and severe necrotic lesions on leaves. Stomatal conductance and photochemical efficiency of photosystem 2 (PhiPS2) of the Nt-cbl10a10b double mutant were significantly inhibited already at a very early stage of the salt stress response. Leaf Na+ concentrations were not much affected in these plants, but the Cl- content of the Nt-cbl10a10b double mutants was significantly lower than that of wild-type plants, which is the first report of CBL10 in the regulation of Cl- homeostasis. Interestingly, the necrosis phenotype of Nt-cbl10a10b double mutants was dependent on light, while the chlorosis phenotype of Nt-cbl10a10b double mutants was light-independent. Different from the previous studies that focus on the role of CBL10 in Na+ homeostasis regulation, this study indicates that NtCBL10 is a key component in regulating multiple aspects of ion homeostasis under salt stress.

Soil salinization poses a mounting global ecological and environmental threat. The identification of genes responsible for negative regulation of salt tolerance and their utilization in crop improvement through gene editing technologies emerges as a swift strategy for the effective utilization of saline-alkali lands. One efficient mechanism of plant salt tolerance is maintaining the proper intracellular K+/Na+ ratio. The Shaker K+ channels play a crucial role in potassium absorption, transport, and intracellular potassium homeostasis in plant cells. Here, the study presents the first genome-wide identification of Shaker K+ channels in Nicotiana tabacum L., along with a detailed bioinformatic analysis of the 20 identified members. Transcriptome analysis revealed a significant up-regulation of NtSKOR1B, an outwardly-rectifying member predominantly expressed in the root tissue of tobacco seedlings, in response to salt stress. This finding was then confirmed by GUS staining of ProNtSKOR1B::GUS transgenic lines and RT-qPCR analysis. Subsequently, NtSKOR1B knockout mutants (ntskor1) were then generated and subjected to salt conditions. It was found that ntskor1 mutants exhibit enhanced salt tolerance, characterized by increased biomass, higher K+ content and elevated K+/Na+ ratios in both leaf and root tissues, compared to wild-type plants. These results indicate that NtSKOR1B knockout inhibits K+ efflux in root and leaf tissues of tobacco seedlings under salt stress, thereby maintaining higher K+/Na+ ratios within the cells. Thus, our study identifies NtSKOR1B as a negative regulator of salt tolerance in tobacco seedlings.

Plant growth and development are driven by intricate processes, with the cell membrane serving as a crucial interface between cells and their external environment. Maintaining balance and signal transduction across the cell membrane is essential for cellular stability and a host of life processes. Ion channels play a critical role in regulating intracellular ion concentrations and potentials. Among these, K+ channels on plant cell membranes are of paramount importance. The research of Shaker K+ channels has become a paradigm in the study of plant ion channels. This study offers a comprehensive overview of advancements in Shaker K+ channels, including insights into protein structure, function, regulatory mechanisms, and research techniques. Investigating Shaker K+ channels has enhanced our understanding of the regulatory mechanisms governing ion absorption and transport in plant cells. This knowledge offers invaluable guidance for enhancing crop yields and improving resistance to environmental stressors. Moreover, an extensive review of research methodologies in Shaker K+ channel studies provides essential reference solutions for researchers, promoting further advancements in ion channel research.

Energy crops play a vital role in meeting future energy and chemical demands while addressing climate change. However, the idealization of low-carbon workflows and careful consideration of cost-benefit equations are crucial for their more sustainable implementation. Here, we propose tobacco as a promising energy crop because of its exceptional water solubility, mainly attributed to a high proportion of water-soluble carbohydrates and nitrogen, less lignocellulose, and the presence of acids. We then designed a strategy that maximizes biomass conversion into bio-based products while minimizing energy and material inputs. By autoclaving tobacco leaves in water, we obtained a nutrient-rich medium capable of supporting the growth of microorganisms and the production of bioproducts without the need for extensive pretreatment, hydrolysis, or additional supplements. Additionally, cultivating tobacco on barren lands can generate sufficient biomass to produce approximately 573 billion gallons of ethanol per year. This approach also leads to a reduction of greenhouse gas emissions by approximately 76% compared to traditional corn stover during biorefinery processes. Therefore, our study presents a novel and direct strategy that could significantly contribute to the goal of reducing carbon emissions and global sustainable development compared to traditional methods.

Salinity stands as a significant environmental stressor, severely impacting crop productivity. Plants exposed to salt stress undergo physiological alterations that influence their growth and development. Meanwhile, plants have also evolved mechanisms to endure the detrimental effects of salinity-induced salt stress. Within plants, Calcineurin B-like (CBL) proteins act as vital Ca2+ sensors, binding to Ca2+ and subsequently transmitting signals to downstream response pathways. CBLs engage with CBL-interacting protein kinases (CIPKs), forming complexes that regulate a multitude of plant growth and developmental processes, notably ion homeostasis in response to salinity conditions. This review introduces the repercussions of salt stress, including osmotic stress, diminished photosynthesis, and oxidative damage. It also explores how CBLs modulate the response to salt stress in plants, outlining the functions of the CBL-CIPK modules involved. Comprehending the mechanisms through which CBL proteins mediate salt tolerance can accelerate the development of cultivars resistant to salinity.

Objectives: Thermal and pH stabilities are extremely important for the application of pectate lyase. In this study, we aimed to obtain a strain that is able to produce pectate lyase with good pH stability.Methods: In this study, screening for pectate lyase was performed using plate assays. Fermentation parameters for pectate lyase production were optimized utilizing a single variable optimization. To get insight into the pectate lyase, its enzyme property, purification, identification, and application were performed.Results: Bacillus altitudinis CAS-WZS-08, producing pectate lyase with good pH stability, was isolated. The optimal fermentation conditions of CAS-WZS-08 are 4 g/L pectin, 20 g/L yeast extract, 2% inoculum size, pH 7.0, and 33 & DEG;C, which the production of pectate lyase can reach up to 0.71 & PLUSMN; 0.001 U/mL. The optimal pH and temperature of the pectate lyase were 10.0 and 60 & DEG;C, respectively. Stored at 4 & DEG;C, the pectate lyase was able to keep its full enzyme activity for 24 h under a wide range of pH (4.0-10.0) condition. With pH 10.0, this pectate lyase was stable under 30-45 & DEG;C. In addition, it can be activated by Mn2+, Cu2+, Co2+, and Ca2+, while inhibited by Fe3+, Ba2+, and Mg2+. Later, the electrophoretic pure protein was acquired through ammonium sulfate precipitation, cation exchange column, and Sephadex G-75. Liquid chromatography tandem-mass spectrometry (LC/MS-MS) further confirmed that the purified protein was pectate lyase with a molecular weight of ti 40 kDa. At last, the result of pectate lyase in extracting apple juice demonstrated that it has an excellent juice extraction ability.Conclusion: This study provides an excellent pH-stable pectate lyase with good thermal stability that is a potential candidate for industrial applications.& COPY; 2023 The Author(s). Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

The CALCINEURIN B-LIKE PROTEIN family genes (CBLs) encode a group of plant-specific calcium sensor proteins that play critical roles in plant development and stress response. Although genome-wide analysis of CBL family genes has been conducted in several diploid plant species, less is known about their functions in allopolyploid plant species. This study conducted a detailed analysis of tobacco CBL family genes. Of the 24 NtCBL genes identified, 14 were derived from the maternal genome donor N. sylvestris, and 10 were from the paternal genome donor N. tomentosiformis. Phylogenetic analysis revealed that NtCBLs could be clustered into three different groups with distinct N-terminal characteristics. All NtCBLs contain EF-hand motifs, but amino acid substitutions and configuration changes may be responsible for variations in their Ca2+-binding abilities. Alternative splicing is common for NtCBL gene transcripts, which may contribute to variations in translation efficiency or protein interaction preferences. Several NtCBL genes exhibited shoot-or root-predominant expression patterns, and some were responsive to salt and/or drought stress. NtCBL4A-1, an ortholog of Arabidopsis thaliana AtCBL4 (SOS3), was predominantly expressed in roots and exhibited a lower gene expression level under salt stress. Further func-tional analysis showed that overexpression of NtCBL4A-1 increased the sensitivity of transgenic tobacco to Na+- induced salt stress, which is inconsistent with the role of AtCBL4 in contributing to salt tolerance. This study provides data for identifying CBL gene functions and selecting candidate stress-tolerant genes in N. tabacum. Additionally, the results may aid gene function studies in other allopolyploid plant species.

Inherently, ammonium (NH4+) is critical for plant growth; however, its toxicity suppresses potassium (K+) uptake and vice-versa. Hence, attaining a nutritional balance between these two ions (NH4+ and K+) becomes imperative for the growth of tobacco seedlings. Therefore, we conducted a 15-day experimental study on tobacco seedlings exposed to different concentrations (47 treatments) of NH4+/K+ at different corresponding 12 ratios simultaneously in a hydroponic system. Our study aimed at establishing the optimal NH4+-K+ concentration and the corresponding ratio required for optimal growth of different tobacco plant organs during the seedling stage. The controls were the baseline for comparison in this study. Plants with low or excessive NH4+-K+ concentration had leaf chlorosis or dark greenish colouration, stunted whole plant part biomass, and thin roots. We found that adequate K+ supply is a pragmatic way to mitigate NH4+-induced toxicity in tobacco plants. The optimal growth for tobacco leaf and root was attained at NH4+-K+ concentrations 2-2 mM (ratio 1:1), whereas stem growth was optimal at NH4+-K+ 1-2 mM (1:2). The study provided an insight into the right combination of NH4+/K+ that could mitigate or prevent NH4+ or K+ stress in the tobacco seedlings.

Xylanase is widely used in various industries such as food processing, paper, textiles, and leather tanning. In this study, Bacillus cereus L-1 strain was isolated and identified as capable of producing low molecular weight xylanase through 16 s rRNA sequencing. Maximum xylanase yield of 15.51 ± 2.08 U/mL was achieved under optimal fermentation conditions (5

Nitrate (NO3-) transporters have been identified as the primary targets involved in plant nitrogen (N) uptake, transport, assimilation, and remobilization, all of which are key determinants of nitrogen use efficiency (NUE). However, less attention has been directed toward the influence of plant nutrients and environmental cues on the expression and activities of NO3- transporters. To better understand how these transporters function in improving plant NUE, this review critically examined the roles of NO3- transporters in N uptake, transport, and distribution processes. It also described their influence on crop productivity and NUE, especially when coexpressed with other transcription factors, and discussed these transporters' functional roles in helping plants cope with adverse environmental conditions. We equally established the possible impacts of NO3- transporters on the uptake and utilization efficiency of other plant nutrients while suggesting possible strategic approaches to improving NUE in plants. Understanding the specificity of these determinants is crucial to achieving better N utilization efficiency in crops within a given environment.

雪茄烟叶中的半纤维素在叶脉及叶片韧性等特性中发挥重要作用,烟叶叶脉较粗,半纤维素含量过高,导致雪茄烟叶韧性较差,可用性变差.本实验室前期筛选到一株产木聚糖酶的蜡样芽孢杆菌,该菌株对烟碱有较好的耐受性,利用液态发酵方法研究菌株以雪茄烟叶为营养源产木聚糖酶的最佳发酵条件,并对最佳发酵条件下菌株降解烟叶中半纤维素的效果进行测定,最后分析比较了菌株对雪茄烟叶香气物质成分及含量的影响.结果表明,菌株在含有不高于1.0 g/L烟碱的培养基中生长受抑制较小,菌株对烟碱有良好的耐受性;当培养基中初始pH 6.7,接种量为5％,雪茄烟叶底物浓度为20 g/L,发酵24 h,菌株产木聚糖酶活性最高,可达(4.04±0.18)U/mL;在最佳发酵条件下,雪茄烟叶中半纤维素含量为3.38％,降低了6.63％,纤维素含量为10.54％,降低了8.19％,利用菌株发酵同时降低了雪茄烟叶中纤维素和半纤维素含量;通过对香气物质影响的分析比较,发现经菌株发酵后的雪茄烟叶能够提升叶绿醇(2.85％)、油酸(0.62％)、正二十六烷(1.75％)和正三十一烷(14.47％)等物质的含量.蜡样芽孢杆菌B.cereus对雪茄烟叶中半纤维素及纤维素降解具有一定作用,后期可应用于雪茄外包皮烟叶的固态发酵,进一步改善雪茄外包皮烟叶的物理特性,为该菌株的开发利用奠定基础.

In order to figure out the effect of organic fertilizers with different carbon-nitrogen (C/N) ratios on the soil improvement and the healthy cultivation, the pot experiment method was used to study effects on the physical and chemical properties and the bacterial community structure of sandy loam soil using five treatments of chemical fertilizer application with the C/N ratios of 15 (CN15), 20 (CN20), 25 (CN25), 30 (CN30) and the control (CK) respectively. Results show that the organic materials with different C/N ratios significantly improve the soil porosity and water content, which all show a linear change rule with the C/N ratio. It can also significantly increase the soil total carbon, total nitrogen, soil C/N ratio, soil microbial biomass carbon, microbial biomass nitrogen and microbial biomass C/N ratio. Among them, CN30 significantly increases the soil total carbon and C/N ratio, which are 5.34-24.13% and 8.87-30.15% respectively compared with other treatments. It can be also found that the dominant flora (at the phylum level) of each treatment are Actinobacteria, Proteobacteria and Chlorobi. The CN30 treatment presents the most obvious improvement in the diversity and richness of the soil bacterial community and is more conducive to the growth and reproduction of Proteobacteria and Firmicutes. The correlation analysis shows that C-total/N-total and C-mic/N-mic are the most important environmental factors affecting the soil physical and chemical properties and their correlation with the bacterial communities. The higher C/N ratio of organic materials results in a more significant improvement of the soil physical and chemical properties. This study provides a new theoretical basis for the soil health cultivation technology.

[背景]烟草在生产和加工中会产生高浓度的尼古丁废弃物,对环境造成较大的污染.[目的]筛选降解尼古丁的微生物菌种并解析其降解尼古丁的代谢途径,理解微生物如何降解尼古丁.[方法]用常规分离筛选方法、结合形态学观察和分子鉴定手段分离和鉴定菌株类别,进而利用单因素试验方法,通过设置不同的尼古丁浓度、温度和pH确定菌株降解尼古丁的最适发酵条件和降解率,利用气相色谱-质谱联用技术检测菌株在尼古丁降解过程中的主要代谢产物.[结果]获得一株以尼古丁为唯一碳源和氮源的节杆菌属(Arthrobacter)菌株,编号为D4;该菌株降解尼古丁的最适温度和pH分别为30.0℃和7.0;在1 g/L的尼古丁浓度下具备较快的尼古丁降解速率,培养18 h时尼古丁降解率可达到90％以上;尼古丁浓度＞4 g/L时菌株生长受到明显抑制;与目前报道的节杆菌属降解途径不同,该菌株降解尼古丁过程中产生了新的终产物N-甲基吡咯烷酮、可替宁及中间产物麦斯明.[结论]本研究分离鉴定到一株具有较快尼古丁降解速率的节杆菌,该菌株很可能存在新的尼古丁降解途径.

Plants have evolved to deal with different stresses during plant growth, relying on complex interactions or crosstalk between multiple signalling pathways in plant cells. In this sophisticated regulatory network, Ca2+ transients in the cytosol ([Ca2+](cyt)) act as major physiological signals to initiate appropriate responses. The CALCINEURIN B-LIKE PROTEIN (CBL)-CBL-INTERACTING PROTEIN KINASE (CIPK) network relays physiological signals characterised by [Ca2+](cyt) transients during plant development and in response to environmental changes. Many studies are aimed at elucidating the role of the CBL-CIPK network in plant growth and stress responses. This review discusses the involvement of the CBL-CIPK pathways in two levels of crosstalk between plant development and stress adaptation: direct crosstalk through interaction with regulatory proteins, and indirect crosstalk through adaptation of correlated physiological processes that affect both plant development and stress responses. This review thus provides novel insights into the physiological roles of the CBL-CIPK network in plant growth and stress adaptation.

Soil organic carbon (SOC), total nitrogen (TN), and their ratio (C:N) play important roles in preserving soil fertility, and their values are closely related to fertilizer use. However, the overall trend and magnitude of changes in SOC, TN and C:N in response to chemical nitrogen fertilizers reduction remain inconclusive. Here, the meta-analysis conducted comparisons at 48 sites covering various cropping system, soil type, and climatic regions of China to investigate the responses of SOC, TN and C:N to chemical nitrogen fertilizers reduction. The results showed that chemical nitrogen fertilizers reduction decreased SOC by 2.76 ± 0.3% and TN by 4.19 ± 0.8%, and increased the C:N by 6.11 ± 0.9% across all the database. Specifically, the reduction of chemical nitrogen without adding organic nitrogen fertilizers would reduce SOC and TN by 3.83% and 11.46% respectively, while they increased SOC and TN by 4.92% and 8.33% respectively with organic fertilizers supplement, suggesting that organic fertilizers could cover the loss of SOC, TN induced by chemical nitrogen fertilizers reduction. Medium magnitude (20–30%) of chemical nitrogen fertilizers reduction enhanced SOC by 6.9%, while high magnitude (≧30%) and total (100%) of chemical nitrogen fertilizers reduction significantly decreased SOC by 3.10% and 7.26% respectively. Moreover, SOC showed a negative response to nitrogen fertilizers reduction at short-term duration (1–2 years), while the results converted under medium-long-termThis system analysis fills the gap on the effects of fertilizer reduction on soil organic carbon and nitrogen at the national scale, and provides technical foundation for the action of reducing fertilizer application while increase efficiency.

Green tea is popularly known for its pleasant flavor and health-care functions. Bitterness and astringency are the two important quality attributes of green tea that enrich tea flavor. Although many research works have focused on the flavor formation of green tea, the review articles about bitterness and astringency is limited. This review article summarizes the major components of bitter and astringent substances in green tea, their sensory perception mechanism, factors influencing the formation of these substances, and the evaluation methods of bitterness and astringency. This review will shed light on the subsequent studies in tea flavor, and provide deeper insight for the research of bitterness and astringency in other foods.

Plants utilize carbohydrates as the main energy source, but much focus has been on the impact of N and K on plant growth. Less is known about the combined impact of NH4+ and K+ nutrition on photoassimilate distribution among plant organs, and the resultant effect of such distribution on growth of tobacco seedlings, hence this study. Here, we investigated the synergetic effect of NH4+ and K+ nutrition on photoassimilate distribution, and their resultant effect on growth of tobacco seedlings. Soluble sugar and starch content peaks under moderate NH4+ and moderate K+ (2-2 mM), leading to improved plant growth, as evidenced by the increase in tobacco weight and root activity. Whereas, a drastic reduction in the above indicators was observed in plants under high NH4+ and low K+ (20-0.2 mM), due to low carbohydrate synthesis and poor photoassimilate distribution. A strong positive linear relationship also exists between carbohydrate (soluble sugar and starch) and the activities of these enzymes but not for invertase. Our findings demonstrated that NH4+ and K+-induced ion imbalance influences plant growth and is critical for photoassimilate distribution among organs of tobacco seedlings.

Many tobacco ( Nicotiana tabacum ) cultivars are salt-tolerant and thus are potential model plants to study the mechanisms of salt stress tolerance. The CALCINEURIN B-LIKE PROTEIN (CBL) is a vital family of plant calcium sensor proteins that can transmit Ca 2+ signals triggered by environmental stimuli including salt stress. Therefore, assessing the potential of NtCBL for genetic improvement of salt stress is valuable. In our studies on NtCBL members, constitutive overexpression of NtCBL5A was found to cause salt supersensitivity with necrotic lesions on leaves. NtCBL5A -overexpressing (OE) leaves tended to curl and accumulated high levels of reactive oxygen species (ROS) under salt stress. The supersensitivity of NtCBL5A -OE leaves was specifically induced by Na + , but not by Cl − , osmotic stress, or drought stress. Ion content measurements indicated that NtCBL5A -OE leaves showed sensitivity to the Na + accumulation levels that wild-type leaves could tolerate. Furthermore, transcriptome profiling showed that many immune response-related genes are significantly upregulated and photosynthetic machinery-related genes are significantly downregulated in salt-stressed NtCBL5A -OE leaves. In addition, the expression of several cation homeostasis-related genes was also affected in salt-stressed NtCBL5A -OE leaves. In conclusion, the constitutive overexpression of NtCBL5A interferes with the normal salt stress response of tobacco plants and leads to Na + -dependent leaf necrosis by enhancing the sensitivity of transgenic leaves to Na + . This Na + sensitivity of NtCBL5A -OE leaves might result from the abnormal Na + compartmentalization, plant photosynthesis, and plant immune response triggered by the constitutive overexpression of NtCBL5A . Identifying genes and pathways involved in this unusual salt stress response can provide new insights into the salt stress response of tobacco plants.

[目的]探索海南H382雪茄烟叶发酵中细菌群落组成和演替规律,为揭示发酵机制提供依据,为筛选功能细菌提供参考.[方法]通过16SrRNA测序技术对海南H382雪茄烟叶不同发酵时期的细菌群落多样性及群落演替规律进行表征,并利用PICRUSt预测了不同发酵时期细菌群落的功能.[结果](1)雪茄烟叶发酵中细菌群落多样性较丰富,7个样本共聚类299个OTU,属于27个属;不同发酵时期样本中OTU数目不同,发酵前期呈增加趋势,随后呈现下降趋势,以YB 3(样本-3)中OTU最多,共222个;(2)以细菌占比多少为标准,H382雪茄烟叶发酵过程中细菌群落演替规律为:含量最高由YB_2中的葡萄球菌属(Staphylococcus)到YB_3中海洋芽孢杆菌属(Oceanobacillus),随后为YB_4(样本-4)中假单胞菌属(Pseudomonas),最后为YB_5、YB_6和YB_7中的葡萄球菌属(Staphylococcus);(3)PICRUSt功能预测结果显示,样本间预测功能种类差异小,而样本间功能种类的丰度差异较大.[结论]H382雪茄烟叶发酵过程中细菌群落组成多样性丰富,优势菌群在本实验条件下是演替的.发酵过程中主要的细菌群落由发酵初期的Staphylococcus、Enterococcus和Pantoea,演变为中期的Oceanobacillus、Paracoccus、Staphylococcus 和 Bacillus,随后演变为Pseudomonas和 Enterobacter,最后稳定为Staphylococcus和Terribacillus.根据细菌的功能预测,它们主要参与氨基酸运输和代谢、碳水化合物的运输和代谢等.

In this study, the possibility of using biochar for soil carbon pool regulation owing to its unique physical and chemical properties was tested. Specifically, to investigate the effects of different biochar particle sizes (500-2000, 250-500, 20-250, and < 20 mu m) on the soil carbon pool structure as well as the associated mechanism, pot culture experiments were performed using peanut shell biochar. The results obtained showed that the chemical properties of the biochar remained unchanged; however, the different particle sizes influenced the porosity properties of the biochar. Smaller-sized biochar particles showed specific surface area (32.91 m2/g), pore volume (0.043 m3/g), and pore size (4.56 nm) that were larger than those corresponding to their largersized counterparts. Further, biochar application enhanced the organic carbon content of the treated soil by up to 46.85 % compared with conventional fertilisation. Biochar with particle size < 20 mu m improved the mean weight diameter of soil water-stable aggregates by 2.5-12.5 %. Furthermore, the easily oxidisable carbon content of the biochar-treated soil reached 2.22 g/kg, increasing by 80.31-89.58 % compared with that achieved via conventional fertilisation. The average soil dissolved organic carbon content was 356.64 mg/kg, which is 78.86 % higher than that achieved via conventional fertilisation. Additionally, biochar with a particle size < 20 mu m also significantly improved the soil carbon pool management index by 186.17 %. Therefore, the results of this study provide a theoretical basis for the scientific application of biochar based on particle size given that micro-particle biochar, in particular, can significantly improve the soil carbon pool management index.