Abstract：
BACKGROUND: To tolerate salt and water-deficit stress, the plant adapts to the adverse environment by regulating its metabolism and expressing certain stress-induced metabolic pathways. This research analyzed the relative expression of four pea genes (P5CR, PAL1, SOD, and POX) in three pea varieties (Climax, Green grass, and Meteor) under different levels of salt and water-deficit stress.
RESULTS: The experiments on salt stress and water-deficit stress were carried out within greenhouse settings under controlled environment. The saturation percentage was employed to create artificial salinity conditions: Control without NaCl treatment, Treatment 1: 50 mM NaCl treatment, Treatment 2: 75 mM NaCl treatment, and Treatment 3: 100 mM NaCl treatment. Field capacity (FC) was used for the development of artificial water-deficit treatments in the pots, i.e., Treatment 1 (Control; water application 100% of FC), Treatment 2 (water application 75% of FC), and Treatment 3 (water application 50% of FC). Pea genes involved in biosynthetic pathways of proline, flavonoids, and enzymatic antioxidant enzymes including P5CR, PAL1, SOD, and POX were selected based on literature. Quantitative real-time PCR using cDNA as a template was used to analyze the gene expression. Pea genes were analyzed for phylogenetic analysis in closely related crops having similarity percent identity 80% and above. In silico characterization of selected proteins including the family classification was done by the NCBI CDD and INTERPRO online servers. Results from RT-qPCR analysis showed increased expression of P5CR, PAL1, and POX genes, while SOD gene expression decreased under both stresses. Climax exhibited superior stress tolerance with elevated expression of P5CR and PAL1, while Meteor showed better tolerance through increased POX expression. Phylogenetic analysis revealed common ancestry with other species like chickpea, red clover, mung bean, and barrel clover, suggesting the cross relationship among these plant species. Conserved domain analysis of respective proteins revealed that these proteins contain PLNO 2688, PLN02457, Cu-Zn Superoxide dismutase, and secretory peroxidase conserved domains. Furthermore, protein family classification indicated that the oxidation-reduction process is the most common chemical process involved in these stresses given to pea plant which validates the relationship of these proteins.
CONCLUSIONS: Salt and water-deficit stresses trigger distinct metabolic pathways, leading to the up-regulation of specific genes and the synthesis of corresponding proteins. These findings further emphasize the conservation of stress-tolerance-related genes and proteins across various plant species. This knowledge enhances our understanding of plant adaptation to stress and offers opportunities for developing strategies to improve stress resilience in crops, thereby addressing global food security challenges.
RESULTS: The experiments on salt stress and water-deficit stress were carried out within greenhouse settings under controlled environment. The saturation percentage was employed to create artificial salinity conditions: Control without NaCl treatment, Treatment 1: 50 mM NaCl treatment, Treatment 2: 75 mM NaCl treatment, and Treatment 3: 100 mM NaCl treatment. Field capacity (FC) was used for the development of artificial water-deficit treatments in the pots, i.e., Treatment 1 (Control; water application 100% of FC), Treatment 2 (water application 75% of FC), and Treatment 3 (water application 50% of FC). Pea genes involved in biosynthetic pathways of proline, flavonoids, and enzymatic antioxidant enzymes including P5CR, PAL1, SOD, and POX were selected based on literature. Quantitative real-time PCR using cDNA as a template was used to analyze the gene expression. Pea genes were analyzed for phylogenetic analysis in closely related crops having similarity percent identity 80% and above. In silico characterization of selected proteins including the family classification was done by the NCBI CDD and INTERPRO online servers. Results from RT-qPCR analysis showed increased expression of P5CR, PAL1, and POX genes, while SOD gene expression decreased under both stresses. Climax exhibited superior stress tolerance with elevated expression of P5CR and PAL1, while Meteor showed better tolerance through increased POX expression. Phylogenetic analysis revealed common ancestry with other species like chickpea, red clover, mung bean, and barrel clover, suggesting the cross relationship among these plant species. Conserved domain analysis of respective proteins revealed that these proteins contain PLNO 2688, PLN02457, Cu-Zn Superoxide dismutase, and secretory peroxidase conserved domains. Furthermore, protein family classification indicated that the oxidation-reduction process is the most common chemical process involved in these stresses given to pea plant which validates the relationship of these proteins.
CONCLUSIONS: Salt and water-deficit stresses trigger distinct metabolic pathways, leading to the up-regulation of specific genes and the synthesis of corresponding proteins. These findings further emphasize the conservation of stress-tolerance-related genes and proteins across various plant species. This knowledge enhances our understanding of plant adaptation to stress and offers opportunities for developing strategies to improve stress resilience in crops, thereby addressing global food security challenges.
摘要：
背景：为了耐受盐和水分亏缺的压力，植物通过调节其代谢和表达某些胁迫诱导的代谢途径来适应不利的环境。本研究分析了四个豌豆基因(P5CR，PAL1,SOD,和POX)在三个豌豆品种(高潮，绿草，和流星）在不同程度的盐和水分亏缺胁迫下。
结果：在受控环境下的温室环境中进行了盐胁迫和水分亏缺胁迫的实验。采用饱和百分比来创建人工盐度条件：未经NaCl处理的对照，治疗1：50mMNaCl治疗，治疗2：75mMNaCl治疗，和处理3:100mMNaCl处理。田间容量（FC）用于在盆中开发人工水分亏缺处理，即，处理1(对照;水应用100%FC),处理2（水应用FC的75％），和处理3(水应用50%的FC)。豌豆基因参与脯氨酸的生物合成途径，黄酮类化合物，和酶抗氧化酶，包括P5CR，PAL1,SOD,和POX是根据文献选择的。使用cDNA作为模板的定量实时PCR来分析基因表达。在具有80%及以上的相似性百分比同一性的密切相关作物中分析豌豆基因用于系统发育分析。通过NCBICDD和INTERPRO在线服务器进行所选蛋白质的计算机表征，包括家族分类。RT-qPCR分析结果显示P5CR的表达增加,PAL1和POX基因，而SOD基因表达在两种胁迫下都降低。Climax表现出优异的胁迫耐受性，P5CR和PAL1表达升高，而Meteor通过增加POX表达表现出更好的耐受性。系统发育分析揭示了与鹰嘴豆等其他物种的共同祖先，红三叶草,绿豆,和桶形三叶草，表明这些植物物种之间的交叉关系。各个蛋白质的保守结构域分析显示，这些蛋白质含有PLNO2688，PLN02457，Cu-Zn超氧化物歧化酶，和分泌过氧化物酶保守结构域。此外，蛋白质家族分类表明，氧化还原过程是豌豆植物受到的这些胁迫中最常见的化学过程，可以验证这些蛋白质之间的关系。
结论：盐和水分亏缺会引发不同的代谢途径，导致特定基因的上调和相应蛋白质的合成。这些发现进一步强调了各种植物物种中与胁迫耐受性相关的基因和蛋白质的保守性。这些知识增强了我们对植物适应压力的理解，并为制定策略以提高作物的抗逆性提供了机会。从而应对全球粮食安全挑战。
结果：在受控环境下的温室环境中进行了盐胁迫和水分亏缺胁迫的实验。采用饱和百分比来创建人工盐度条件：未经NaCl处理的对照，治疗1：50mMNaCl治疗，治疗2：75mMNaCl治疗，和处理3:100mMNaCl处理。田间容量（FC）用于在盆中开发人工水分亏缺处理，即，处理1(对照;水应用100%FC),处理2（水应用FC的75％），和处理3(水应用50%的FC)。豌豆基因参与脯氨酸的生物合成途径，黄酮类化合物，和酶抗氧化酶，包括P5CR，PAL1,SOD,和POX是根据文献选择的。使用cDNA作为模板的定量实时PCR来分析基因表达。在具有80%及以上的相似性百分比同一性的密切相关作物中分析豌豆基因用于系统发育分析。通过NCBICDD和INTERPRO在线服务器进行所选蛋白质的计算机表征，包括家族分类。RT-qPCR分析结果显示P5CR的表达增加,PAL1和POX基因，而SOD基因表达在两种胁迫下都降低。Climax表现出优异的胁迫耐受性，P5CR和PAL1表达升高，而Meteor通过增加POX表达表现出更好的耐受性。系统发育分析揭示了与鹰嘴豆等其他物种的共同祖先，红三叶草,绿豆,和桶形三叶草，表明这些植物物种之间的交叉关系。各个蛋白质的保守结构域分析显示，这些蛋白质含有PLNO2688，PLN02457，Cu-Zn超氧化物歧化酶，和分泌过氧化物酶保守结构域。此外，蛋白质家族分类表明，氧化还原过程是豌豆植物受到的这些胁迫中最常见的化学过程，可以验证这些蛋白质之间的关系。
结论：盐和水分亏缺会引发不同的代谢途径，导致特定基因的上调和相应蛋白质的合成。这些发现进一步强调了各种植物物种中与胁迫耐受性相关的基因和蛋白质的保守性。这些知识增强了我们对植物适应压力的理解，并为制定策略以提高作物的抗逆性提供了机会。从而应对全球粮食安全挑战。
