Dataset for "High photosynthesis rates in Brassiceae species are mediated by leaf anatomy enabling high biochemical capacity, rapid CO2 diffusion and efficient light use"

Dataset used in the manuscript 'High photosynthesis rates in Brassiceae species are mediated by leaf anatomy enabling high biochemical capacity, rapid CO2 diffusion and efficient light use'. Please cite the paper presenting this datase. Plant Species and Inbred Lines: Hirschfeldia incana L. (7th generation inbred line 190003 HIN-NIJ-07-B) Brassica nigra L. (3rd generation inbred line 210093 BNI-DG1-03-B) Brassica rapa L. (inbred line ‘R-o-18’) Arabidopsis thaliana (accession Columbia) Growth Conditions: Media: Rock-wool blocks (Grodan Plantop, Roermond, Netherlands, 10×10×7.5 cm) Fertigation: Nitrogen-rich nutrient solution via automated dripping system. Light Conditions: 12 h day/12 h night, light intensity of 200 µmol m-2 s-1 and 1800 µmol m-2 s-1 Temperature: Day/Night temperatures of 23 °C and 20 °C, respectively. Relative Humidity: 70% Codes Species: Hirschfeldia incana L. - H. incana Brassica nigra L. - B. nigra Brassica rapa L. - B. rapa Arabidopsis thaliana - A. thaliana Light conditions: High light - HL Low light - LL Replicates: Biological replicates were labeled with numbers, e.g. replicate one from Hirschfeldia incana is referred to as HiHL1 Measurements Leaf Gas Exchange and Chlorophyll Fluorescence Measurements (GasExchangeData.zip): Four leaves per species per treatment. Conducted using a LI-6800 (LI-COR, Lincoln, NE, USA) on the mid-position of the youngest fully expanded leaf. The resoponse of photosynthesis to irradiance and external CO2 concentrations augumneted with multi-phase flash fluorescence were made Optical Properties Measurement (Absorbance & chlorophyll.zip): Leaves: Four leaves per species per treatment. Leaf transmittance and reflectance measured using a dual channel spectrophotometer (absorptance_reflac_data_355_750.xlsx) Chlorophyll content measured using a spectrophotometer (Chlorophyll.xlsx). Stomatal Density and Size Analysis (Stomata.zip): Sampling: Leaves: Four leaves per species per treatment. Plants: Samples taken from three different plants. Leaf-side: abaxial and adxial leaf side. Microscopy Setup: Stomatal imprints made using clear nail polish, imaged using a light microscope at 20x. Data Output: Imaging Results: .jpg files organised under folders for species e.g. AtHL\R1 T+B.zip contains images ofimprints of top (T) and bottom (B) leaf sides from replicate plant 1 (R1) of A. thaliana grown under high light (AtHL). The images are named as for example, AT_HL_BOTTOM_R1_A_stacked_minimum.jpg, The leters A to E label various imges made from one imprint. Light and Electron Microscopy of Leaf Sections (CellwallChloroplast.zip): Sampling: Leaves: Four leaves per species per treatment. Plants: Samples taken from four different plants. Sample preparation Leaf samples fixed, dehydrated, embedded in Araldite, and sectioned for imaging. 1 µm think sections were made for light microscopy Sections of 70 nm were double stained for TEM Microscopy Setup: Mesophyll cells imaged at 400x and 700x to measure chloroplast coverage. Electron microscopy performed with Zeiss EM900 electron microscope. Mesophyll Chlorophyll (ConfocalData.zip): Sampling: Leaves: Three leaves per species per treatment. Plants: Samples taken from three different plants. Thickness: 200 ± 10 µm sections prepared using a sliding microtome Microscopy Setup: Microscope: Leica DM8 inverted scope equipped with a Stellaris 5 confocal microscope (Leica Microsystems, Wetzlar, Germany). Excitation: 490 nm excitation laser line Fluorescence Recording: Chlorophyll autofluorescence recorded in a spectral range of 660−700 nm. Objective: Leica objective ×10/0.4 NA. Z-Stacks: 85–112 µm depth, two random positions per sample Data Output: Imaging Results: Z-stacks of chlorophyll autofluorescence in mesophyll cells. Spectral Information: Chlorophyll autofluorescence recorded in the 660−700 nm range. Analysis: Software: The confocal files are in .lif format and can be viewed using Leica applixation suite (LASx), ImageJ

本数据集用于论文《十字花科物种的高光合速率由叶片解剖结构介导，具备高生化容量、快速CO₂扩散与高效光能利用》，请引用提供本数据集的相关论文。

## 植物物种与自交系
野萝卜（Hirschfeldia incana L.，第7代自交系190003 HIN-NIJ-07-B）、黑芥（Brassica nigra L.，第3代自交系210093 BNI-DG1-03-B）、白菜型油菜（Brassica rapa L.，自交系‘R-o-18’）与拟南芥（Arabidopsis thaliana，哥伦比亚生态型Columbia）。

## 培养条件
1. 基质：采用岩棉块（Grodan Plantop，荷兰鲁尔蒙德，尺寸10×10×7.5 cm）
2. 灌溉施肥：通过自动滴灌系统施用富氮营养液
3. 光照条件：设置12 h昼长/12 h夜长，光照强度分别为200 µmol m⁻² s⁻¹与1800 µmol m⁻² s⁻¹
4. 温度：昼/夜温度分别为23 ℃与20 ℃
5. 相对湿度：70%

## 编码规则
1. 物种缩写：Hirschfeldia incana L. → H. incana；Brassica nigra L. → B. nigra；Brassica rapa L. → B. rapa；Arabidopsis thaliana → A. thaliana
2. 光照组别：高光（High light, HL）、低光（Low light, LL）
3. 生物学重复：以数字标注，例如野萝卜高光处理下的重复1记为HiHL1

## 测定项目
### 叶片气体交换与叶绿素荧光测定（GasExchangeData.zip）
每个物种每个处理组取4片叶片，使用LI-6800便携式光合仪（LI-COR，美国内布拉斯加州林肯市）于完全展开幼叶的中部位置进行测定，内容包括光合速率对光照强度与外界CO₂浓度的响应，并辅以多相闪光荧光技术。

### 光学特性测定（Absorbance & chlorophyll.zip）
每个物种每个处理组取4片叶片：使用双通道分光光度计测定叶片透射率与反射率（数据文件：absorptance_reflac_data_355_750.xlsx）；使用分光光度计测定叶绿素含量（数据文件：Chlorophyll.xlsx）。

### 气孔密度与大小分析（Stomata.zip）
采样方案：每个物种每个处理组取4片叶片，样本取自3株不同植株，分别采集叶背（abaxial）与叶面（adaxial）的样本。制样与成像：使用透明指甲油制作气孔印迹，通过光学显微镜20×物镜成像。数据输出：成像结果为.jpg格式文件，按物种分组存储于对应文件夹，例如AtHLR1 T+B.zip包含高光处理下拟南芥（A. thaliana，简称At）重复1（R1）植株的叶上（T）、叶下（B）面气孔印迹图像。图像命名示例：AT_HL_BOTTOM_R1_A_stacked_minimum.jpg，其中A至E字母标注同一张印迹的不同成像结果。

### 叶片切片光学与电子显微镜观察（CellwallChloroplast.zip）
采样方案：每个物种每个处理组取4片叶片，样本取自4株不同植株。样本制备：固定叶片样本，脱水后包埋于环氧树脂Araldite中，制作1 µm厚半薄切片用于光学显微镜观察；制作70 nm超薄切片，经双重染色后用于透射电子显微镜（Transmission Electron Microscope, TEM）观察。显微镜设置：使用400×与700×物镜拍摄叶肉细胞以测定叶绿体覆盖度；使用Zeiss EM900电子显微镜完成电子显微成像。

### 叶肉叶绿素荧光成像（ConfocalData.zip）
采样方案：每个物种每个处理组取3片叶片，样本取自3株不同植株。切片制备：使用滑动式切片机制作厚度为200 ± 10 µm的切片。显微镜设置：使用配备Stellaris 5共聚焦模块的Leica DM8倒置显微镜（Leica Microsystems，德国韦茨拉尔）；激发光源采用490 nm激光线；在660−700 nm光谱范围内记录叶绿素自发荧光；使用Leica ×10/0.4 NA物镜成像；Z-stack成像深度为85–112 µm，每个样本采集2个随机位置的图像序列。数据输出：成像结果为叶肉细胞叶绿素自发荧光的Z-stack序列图像，同时采集660−700 nm范围内的叶绿素自发荧光光谱信息。分析软件：共聚焦文件格式为.lif，可通过Leica应用套件（LASx）与ImageJ软件查看。