Li Jiajia, Liu Xinyu, Xu Lingqing, Li Wangsheng, Yao Qi, Yin Xilong, Wang Qiuhong, Tan Wenbo, Xing Wang, Liu Dali
National Beet Medium-term Gene Bank, Heilongjiang University, Harbin, China.
Key Laboratory of Sugar Beet Genetics and Breeding, Heilongjiang Province Common College/College of Advanced agriculture and ecological environment, Heilongjiang University, Harbin, China.
Front Plant Sci. 2023 Apr 21;14:1164151. doi: 10.3389/fpls.2023.1164151. eCollection 2023.
Nitrogen (N) is an essential macronutrient for plants, acting as a common limiting factor for crop yield. The application of nitrogen fertilizer is related to the sustainable development of both crops and the environment. To further explore the molecular response of sugar beet under low nitrogen (LN) supply, transcriptome analysis was performed on the LN-tolerant germplasm '780016B/12 superior'. In total, 580 differentially expressed genes (DEGs) were identified in leaves, and 1,075 DEGs were identified in roots (log ≥ 1; q value < 0.05). Gene Ontology (GO), protein-protein interaction (PPI), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses clarified the role and relationship of DEGs under LN stress. Most of the downregulated DEGs were closely related to "photosynthesis" and the metabolism of "photosynthesis-antenna proteins", "carbon", "nitrogen", and "glutathione", while the upregulated DEGs were involved in flavonoid and phenylalanine biosynthesis. For example, (glutamate dehydrogenase B) was identified as a key downregulated gene, linking carbon, nitrogen, and glutamate metabolism. Thus, low nitrogen-tolerant sugar beet reduced energy expenditure mainly by reducing the synthesis of energy-consuming amino acids, which in turn improved tolerance to low nitrogen stress. The glutathione metabolism biosynthesis pathway was promoted to quench reactive oxygen species (ROS) and protect cells from oxidative damage. The expression levels of nitrogen assimilation and amino acid transport genes, such as (high-affinity nitrate transporter), (nitrate reductase [NADH]), (ferredoxin-nitrite reductase), (glutamine synthetase leaf isozyme), (glutathione transferase) and (glutathione hydrolase 3) at low nitrogen levels play a decisive role in nitrogen utilization and may affect the conversion of the carbon skeleton. (dihydroflavonol 4-reductase) in roots was negatively correlated with in leaves (coefficient = -0.98, p < 0.05), suggesting that there may be corresponding remote regulation between "flavonoid biosynthesis" and "nitrogen metabolism" in roots and leaves. (fructose 1,6-bisphosphatase) and (phosphoglycerate kinase) were significantly positively correlated (p < 0.001) with Ci (intercellular CO concentration). The reliability and reproducibility of the RNA-seq data were further confirmed by real-time fluorescence quantitative PCR (qRT-PCR) validation of 22 genes (R = 0.98). This study reveals possible pivotal genes and metabolic pathways for sugar beet adaptation to nitrogen-deficient environments.
氮(N)是植物必需的大量营养素,是作物产量的常见限制因素。氮肥的施用与作物和环境的可持续发展相关。为了进一步探究低氮(LN)供应条件下甜菜的分子响应,对耐低氮种质‘780016B/12优系’进行了转录组分析。总共在叶片中鉴定出580个差异表达基因(DEGs),在根中鉴定出1075个DEGs(log≥1;q值<0.05)。基因本体论(GO)、蛋白质-蛋白质相互作用(PPI)和京都基因与基因组百科全书(KEGG)分析阐明了低氮胁迫下DEGs的作用和关系。大多数下调的DEGs与“光合作用”以及“光合天线蛋白”“碳”“氮”和“谷胱甘肽”的代谢密切相关,而上调的DEGs参与类黄酮和苯丙氨酸生物合成。例如,谷氨酸脱氢酶B被鉴定为关键的下调基因,它连接碳、氮和谷氨酸代谢。因此,耐低氮甜菜主要通过减少耗能氨基酸的合成来降低能量消耗,进而提高对低氮胁迫的耐受性。谷胱甘肽代谢生物合成途径被促进以淬灭活性氧(ROS)并保护细胞免受氧化损伤。低氮水平下氮同化和氨基酸转运基因的表达水平,如高亲和硝酸盐转运蛋白、硝酸还原酶(NADH)、铁氧还蛋白-亚硝酸还原酶、谷氨酰胺合成酶叶片同工酶、谷胱甘肽转移酶和谷胱甘肽水解酶3在氮利用中起决定性作用,可能影响碳骨架的转化。根中的二氢黄酮醇4-还原酶与叶片中的呈负相关(系数=-0.98,p<0.05),表明根和叶中“类黄酮生物合成”与“氮代谢”之间可能存在相应的远程调控。果糖1,6-二磷酸酶和磷酸甘油酸激酶与胞间CO浓度显著正相关(p<0.001)。通过对22个基因的实时荧光定量PCR(qRT-PCR)验证进一步证实了RNA-seq数据的可靠性和可重复性(R=0.98)。本研究揭示了甜菜适应缺氮环境可能的关键基因和代谢途径。