彩色小麦富含花青素和矿质元素，具有独特的营养功能，在消费市场获得广泛青睐。然而，紫色小麦的高花青素品种常伴随产量低制约其产业发展。氮肥和密度是影响小麦产量的重要栽培措施，增加施氮量可提高紫色小麦籽粒产量，但会降低花青素合成影响其品质。适当提高种植密度，可有效补偿减氮导致的产量下降。本研究选用宁紫1933品种为材料，于2022-2023和2023-2024年度在句容和姜堰两个生态点开展大田试验。试验设计如下：设置氮密互作处理（第一年度氮肥水平：180、240和300 kg ha^-1；第二年度调整氮肥水平为：120、180和240 kg ha^-1；两年度相同的种植密度：240、300和360万株 ha^-1），本研究系统分析了两个生长年度中氮密互作对产量形成和品质的调控效应，深入解析了减少施氮量和增加种植密度调控籽粒花青素合成的生理机制。主要研究结果如下：

1. 氮密互作对紫色小麦生长发育、产量和花青素含量的影响

减少施氮量显著降低了紫色小麦开花期至花后30天（30DAA）旗叶叶绿素相对含量（SPAD）以及植株叶面积指数（LAI），适量增加种植密度可以提高紫色小麦的LAI。同时，种植密度的增加也提高了拔节期茎糵数和成熟期穗数，从而缓解了因为氮肥减少而造成的产量损失。与对照N₂₄₀D₂₄₀相比，高氮水平下时籽粒花青素含量减少，籽粒种皮的颜色（强度I值）显著降低，随着氮肥的减少和密度的增加，籽粒花青素含量和强度I值不断提高。同时籽粒中氮含量、氮积累量随氮肥的减少和密度的增加显著降低，分别与花青素含量、花青素积累量呈负相关关系。减少施氮量和增加种植密度显著促进了籽粒中花青素的含量（与对照相比，句容试验点中第一年度最高增加了124.7%，第二年度达156.4%）。在姜堰试验点，减少施氮量和增加种植密度对于花青素含量的调控效应较句容小，与对照相比最高增加了33.2%。

2. 氮密互作对紫色小麦花青素合成生理机制的影响

灌浆期籽粒花青素积累动态呈先上升后下降的趋势，花后15天（15DAA）开始积累，30DAA时达到峰值，随后有所下降，在成熟期时达到稳定。相较对照，N₃₀₀时花青素积累速率降低了7.0%，N₁₈₀时花青素积累速率增加了12.0%，增加种植密度使得花青素积累速率略有提高。在紫色小麦籽粒中共检测到15种相对含量较高的花青素组分。20DAA至30DAA主要增加了各类花青素酰基化衍生物的相对含量，30DAA至成熟期主要是矢车菊素和飞燕草素相对含量的降低。当施氮量由240 kg ha^–1减至120 kg ha^-1、种植密度由240万株 ha^-1增至360万株 ha^-1，主要增加了矢车菊素、飞燕草素及芍药花素组分的含量，结构基因中类黄酮-3'-羟化酶（F3'H）、类黄酮-3-葡萄糖基转移酶（UFGT）和二氢黄酮醇-4-还原酶（DFR）基因的相对表达水平有大幅提高，其次为黄烷酮-3-羟化酶（F3H）、查尔酮合成酶（CHS）、类黄酮-3'5'-羟化酶（F3'5'H）、花青素合成酶（ANS）、查尔酮异构酶（CHI）基因相对表达水平的少量提高，同时调节基因TaPpm1、TaPpb1和MYC4E的相对表达水平也有提高。

3. 氮密互作对紫色小麦营养和加工品质的影响

发现N₃₀₀水平时，籽粒类黄酮含量较对照降低了5.4%，总酚和抗氧化能力的差异不显著。当施氮量由300 kg ha^–1减至180 kg ha^-1、种植密度由240万株 ha^-1增至360万株 ha^-1，籽粒总酚、类黄酮含量和抗氧化能力均显著增加，同时增加了P、K、Mn、Zn和Se等矿质元素的含量。在2022-2023年句容试验点中，减少氮肥和增加种植密度的处理均降低了籽粒蛋白质和湿面筋的含量，提高了淀粉含量，降低了面粉筋力，提高了淀粉的糊化特性和热稳定性，缩短了糊化时间，使得全麦粉的六大指标分布图接近于馒头目标指数剖面图，增加了全麦馒头的比容、孔隙率、气孔密度、弹性、回复性和粘聚性，降低了宽高比、硬度、咀嚼度、胶着性和平均气孔面积，感官评价总分达到89分。

上述结果揭示了氮密互作对紫色小麦产量和花青素含量的调控规律，从氮素水平、植株生长发育及分蘖特性方面阐明了氮密互作对籽粒产量和花青素含量形成的协同调控作用；从花青素组分和花青素合成基因的相对表达水平角度解析了氮密互作促进花青素合成的代谢通路；氮密互作改善籽粒中生物活性物质和矿质元素等营养品质的含量，并通过调控蛋白质和湿面筋含量改善了紫色小麦的加工品质，实现了馒头烘焙品质的显著提升。这些研究成果不仅为紫色小麦优质高产栽培提供了重要的理论依据，也为实际生产中的技术选择提供了科学指导。

Colored wheat is rich in anthocyanins and mineral elements and has unique nutritional functions, thus gaining wide favor in the consumer market. However, purple wheat varieties with high anthocyanin content are often accompanied by a decrease in yield. which restricts the development of its industry.Nitrogen fertilizer and density are important cultivation measures that affect purple wheat yield. Increasing the amount of nitrogen application can enhance the grain yield of wheat, but it will reduce anthocyanin synthesis and affect the quality of colored wheat. Appropriately increasing the planting density can effectively compensate for the decline in yield caused by nitrogen reduction. This study selected Ningzi 1933 varieties as materials and conducted field trials in two ecological sites, Jurong and Jiangyan, during the 2022-2023 and 2023-2024 years. The experimental design is as follows: Nitrogen and density interaction treatment was set up (Nitrogen fertilizer levels in the first year: 180, 240 and 300 kg ha^-1; Nitrogen fertilizer levels in the second year was adjusted to: 120, 180 and 240 kg ha^-1; With the same planting densities in two years: 2.4, 3 and 3.6 million seedlings ha^-1), this study systematically analyzed the regulatory effects of nitrogen and density interaction on yield formation and quality in two growth years, deeply analyzed the physiological mechanism by which reducing nitrogen application rate and increasing the planting density regulates anthocyanin synthesis in grains. The main research results are as follows:

1. Effects of nitrogen and density interaction on the growth and development, yield and anthocyanin content of purple wheat

Reducing the nitrogen application rate significantly decreased the chlorophyll relative content (SPAD) of flag leaves and the leaf area index (LAI) of plants in purple wheat from the flowering period to 30 days after flowering. Moderately increasing the planting density can improve the LAI of purple wheat. Meanwhile, the increased planting density also raised the tillering number and percentage of earbearing tiller, thereby alleviating the yield loss caused by the reduced nitrogen fertilizer. Compared with the control (N₂₄₀D₂₄₀), the anthocyanin content in grains decreased at high nitrogen levels, and the color (intensity I value) of the seed coat of grains significantly decreased. With the reduction of nitrogen fertilizer and the increase of density, the anthocyanin content and intensity I value in grains continuously increased. Meanwhile, the nitrogen content and nitrogen accumulation in the grains decreased significantly with the reduction of nitrogen fertilizer and the increase of density. It was negatively correlated with the anthocyanin content and the accumulation of anthocyanin respectively. Reducing the nitrogen application rate and increasing the planting density significantly promoted the anthocyanin content in the grains (compared with the control, in the Jurong test site, it increased by up to 124.7% in the first year and 156.4% in the second year). At the Jiangyan experimental site, the regulatory effects of reducing nitrogen application rate and increasing the planting density on anthocyanin content were smaller than those in Jurong, and increased by up to 33.2% compared with the control.

2. Effect of nitrogen and density interaction on the physiological mechanism of anthocyanin synthesis in purple wheat.

 During the grain filling period, the accumulation dynamics of anthocyanins in grains showed a trend of first increasing and then decreasing. Accumulation began at 15DAA, reached the peak at 30DAA, then decreased, and stabilized at the mature stage. Compared with the control, the accumulation rate of anthocyanin decreased by 7.0% at N₃₀₀, increased by 12.0% after N₁₈₀. The increase in planting density slightly increased the accumulation rate of anthocyanin. A total of 15 anthocyanin components with relatively high contents were detected in the grains of purple wheat.From 20DAA to 30DAA, the relative contents of various anthocyanin acylated derivatives mainly increased. From 30DAA to the mature stage, the relative contents of cyanidin and delphinidin mainly decreased. When the nitrogen application rate was reduced from 240 kg ha^-1 to 120 kg ha^-1 and the planting density was increased from 2.4 million seedlings ha^-1 to 3.6 million seedlings ha^-1, the contents of cyanidin, delphinidin and peonidin components were mainly increased. The relative expression levels of flavonoid-3'-hydroxylase (F3'H), flavonoid-3-glucosyltransferase (UFGT), and dihydroflavonol-4- reductase (DFR) genes in structural genes have been significantly increased. Secondly, there is an increase in the relative expression levels of a small amount of flavanone-3-hydroxylase (F3H), chalcone synthase (CHS), flavonoid 3'5'-hydroxylase (F3'5'H), anthocyanin synthase (ANS), and chalcone isomerase (CHI) genes. Meanwhile, the relative expression levels of the regulatory genes TaPpm1, TaPpb1 and MYC4E also increased.

3. Effect of nitrogen and density interaction on the nutrition and processing quality of purple wheat.

When the N₃₀₀ level was found, the flavonoid content decreased by 5.4% compared with the control, while the differences in total phenols and antioxidant capacity were not significant. When the nitrogen application rate was reduced from 300 kg ha^-1 to 120 kg ha^-1 and the planting density was increased from 2.4 million seedlings ha^-1 to 3.6 million seedlings ha^-1, the contents of total phenols and flavonoids in the grains and the antioxidant capacity all increased significantly, while the contents of mineral elements such as P, K, Mn, Zn and Se were increased. In the Jurong test site from 2022 to 2023, the treatments of reducing nitrogen fertilizer and increasing planting density both decreased the contents of grain protein and wet gluten, increased the starch content, reduced the gluten strength of flour, improved the gelatinization characteristics and thermal stability of starch, shortened the gelatinization time, and made the distribution map of the six major indicators of whole wheat flour close to the cross-sectional diagram of the target index of steamed bread. The specific volume, poriness, stomotal density, springness, resilience and cohesiveness of whole wheat steamed bread, reduces the aspect ratio, hardness, chewiness, gumminess and mean stomatal area, and the total score of sensory evaluation reaches 89 points.

The above results reveal the regulatory laws of nitrogen and density interaction on the yield and anthocyanin content of purple wheat, and clarify the synergistic regulatory effect of nitrogen and density interaction on the formation of grain yield and anthocyanin content from the aspects of nitrogen level, plant growth and development, and tillering characteristics. The metabolic pathways through which nitrogen and density interaction promotes anthocyanin synthesis were analyzed from the perspective of the relative expression levels of anthocyanin components and anthocyanin synthesis genes. It was clarified that the nitrogen and density interaction regulated the contents of nutrients such as bioactive substances and mineral elements in the grains, and improved the processing quality of purple wheat by regulating the contents of protein and wet gluten, achieving a significant improvement in the baking quality of steamed bread. These research results not only provide important theoretical basis for the high-quality and high-yield cultivation of purple wheat, but also offer scientific guidance for the technical selection in actual production.

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