基于叶片显微结构筛选耐盐碱苹果砧木_许盼云.pdf

研究报告Research Report基于叶片显微结构筛选耐盐碱苹果砧木许盼云李春兰马萍刘畅何天明*新疆农业大学林学与园艺学院,乌鲁木齐,830052*通信作者,摘要从叶片解剖学角度分析不同苹果砧木品种的耐盐碱性，旨在筛选适宜新疆南疆盐碱土壤栽培的优良苹果砧木。本研究以 7 种四年生苹果砧木为试验材料，将 NaCl、Na2SO4、NaHCO3、Na2CO3按 1:9:9:1 的比例混合，盆栽浇灌 1/2 Hoagland 营养液，模拟浓度为 60 mmol/L 的盐碱复合胁迫，对照浇灌 1/2 Hoagland 营养液。通过光学显微镜观察 14 项叶片解剖结构，并结合主成分分析法和聚类系统法综合评价各砧木的耐盐碱性。研究发现盐碱复合胁迫对砧木叶片叶肉解剖结构产生的影响大于叶脉结构；盐碱复合胁迫后，耐盐碱性较强的砧木叶片的栅栏组织细胞、维管束结构和厚薄壁组织细胞变得发达，组织紧实度增大，而耐盐碱性较弱的砧木叶片的栅栏组织细胞短缩，海绵组织疏松，组织疏松度变大，主脉厚度减小。结果表明，7 种供试材料的耐盐碱性分为三类，包括耐盐碱性强的八棱海棠，较强的小金海棠和平邑甜茶，较弱的野苹果、白海棠、山定子、湖北海棠。本研究结果为耐盐碱性强的苹果砧木选育提供一定的理论依据和参考价值。关键词苹果砧木;叶片解剖结构;耐盐碱性;综合评价Screening of Salt-Alkali Tolerant Apple Rootstocks Based on Leaf Mi-crostructureXu PanyunLi ChunlanMa PingLiu ChangHe Tianming*College of Forestry and Horticulture,Xinjiang Agricultural University,Urumqi,830052*Corresponding author,DOI:10.13271 j.mpb.021.000950AbstractThe salt-alkali tolerance of different apple rootstocks was analyzed from the perspective of leaf anato-my in order to select the superior apple rootstocks suitable for salt-alkali soil cultivation in Southern Xinjiang.Using 7 four-year-old apple rootstocks as test materials,mix NaCl,Na2SO4,NaHCO3,and Na2CO3at a ratio of 1:9:9:1,and water the pot with 1/2 Hoagland nutrient solution,with a simulated concentration of 60 mmol/L saline alkalicompound stress,the control was poured 1/2 Hoagland nutrient solution.14 anatomical structures of leaves wereobserved by optical microscope,and the salt-alkali tolerance of each rootstock was evaluated by principal compo-nent analysis and clustering system method.The study found that the salt-alkali complex stress has greater influ-ence on the anatomical structure of the leaf flesh of the rootstock than the vein structure.After salt-alkali complexstress,palisade tissue cells,vascular bundle structure and parenchyma tissue cells of strong salt-alkali tolerancerootstock leaves became developed and tissue compactness increased,while palisade tissue cells of weak salt-alkalitolerance rootstock leaves shortened,spongy tissue became loose,tissue looseness increased and main vein thick-ness decreased.The results showed that the salt-alkali tolerance of the 7 tested materials could be divided intothree categories,including the strong salt-alkali tolerance of M.robusta,the stronger M.xiaojinensis and M.hupe-基金项目：本研究由国家公益性行业(农业)科研专项(20130309305)资助引用格式：Xu P.Y.,Li C.L.,Ma P.,Liu C.,and He T.M.,2023,Screening of salt-alkali tolerant apple rootstocks based on leaf mi-crostructure,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),21(3):950-957.(许盼云,李春兰,马萍,刘畅,何天明,2023,基于叶片显微结构筛选耐盐碱苹果砧木,分子植物育种,21(3):950-957.)分子植物育种，2023 年，第 21 卷，第 3 期，第 950-957 页Molecular Plant Breeding,2023,Vol.21,No.3,950-957图 1 苹果砧木品种盐碱复合胁迫前后叶片横切面显微结构变化注:放大倍数:20 倍;比例尺:100 m;A,E 分别代表平邑甜茶和湖北海棠在 0 mmol/L 盐碱复合胁迫前叶片横切面显微结构;B,F 分别代表平邑甜茶和湖北海棠在 60 mmol/L 盐碱复合胁迫后叶片横切面显微结构;C,G 分别代表平邑甜茶和湖北海棠在 0 mmol/L 盐碱复合胁迫前叶片主脉横切面显微结构;D,H 分别代表平邑甜茶和湖北海棠在 60 mmol/L 盐碱复合胁迫后叶片主脉横切面显微结构;UE:上表皮;LE:下表皮;PT:栅栏组织;ST:海绵组织;X:木质部;Ph:韧皮部;VB:主脉维管束;TP:厚薄壁组织(厚角和薄壁组织)Figure 1 Changes of leaf cross-section microstructure before andafter salt-alkali compound stress in apple rootstock varietiesNote:Magnification:20 times;Scale:100 m;A,E respectivelyrepresent the microstructure of leaf cross-section of M.hupehensis var.pingyiensis and M.hupehensis.before 0 mmol/L salt-al-kali complex stress;B,F respectively represent the microstructureof leaf cross-section of M.hupehensis var.pingyien-sis and M.hupehensis under 60 mmol/L salt-alkali complex stress;C,G re-spectivelyrepresent the microstructure of leaf main vein cross-sec-tion of M.hupehensis Rehd.and M.hupehensis(Pamp.)Rehder.before 0 mmol/L salt-alkali complex stress;D,H respectively rep-resent the microstructure ofleafmain vein cross-section of M.hu-pehensis var.pingyiensis and M.hupehensis 60 mmol/L salt-al-kali complex stress;UE:Upper epidermis;LE:Lower epidermis;PT:Palisade tissue;ST:Sponge tissue;X:Xylem;Ph:Phloem;VB:Vascular bundle;TP:Thick parenchyma(thick angles andparenchyma)叶片是植物体进行光合作用、气体交换及蒸腾作用最主要的营养器官，生境的改变会使叶片形态和微观解剖结构发生相应的变化。在一定条件下，叶片结构也反映了植物对环境的适应能力(Chen et al.,2014)。因此，检测叶片解剖结构成为评估植物抗性的一种简单而可靠的方法(马婷等,2016;木巴热克 阿尤普等,2019)。刘育梅等(2017)利用盆栽试验对 5 种山榄科树种叶片组织结构与耐盐性之间的关系检测和分析，发现耐盐树种的气孔器密度较小，并且叶片、下角质层、上表皮和栅栏组织比不耐盐树种的厚度显著增厚。混合盐碱胁迫下，垂丝海棠可显著增加叶片厚度，从而减少水分蒸发，增加叶片保水能力，以提高耐盐性(朱燕芳,2018)。本研究通过人为模拟新疆南疆地区 pH 8.83 的盐碱土，通过对叶片显微结构进行分析，检测不同苹果砧木品种的耐盐碱性，对各砧木成熟叶片的叶肉和主脉显微结构进行观测，从各项解剖结构指标在盐碱复合胁迫环境的变化来分析，利用主成分分析和聚类分析方法对其耐盐碱性进行综合排序和分类，旨在筛选适宜新疆南疆盐碱土壤栽培的苹果砧木，为耐盐碱性强的苹果砧木选育提供一定的参考价值。1结果与分析1.1不同苹果砧木品种叶片的解剖结构分析以平邑甜茶和湖北海棠为例，盐碱复合胁迫前后叶片横切面显微结构显示(图 1)，苹果叶片为典型的异面叶。上下表皮由近方形或矩形的 1 层细胞组成，且下表皮伴有气孔分布；栅栏组织垂直于表皮细胞，长圆柱状，密集排列呈栅状，常为 23 层，组织中包含较多的叶绿体；下表皮附近是海绵组织，细胞形状多不规则，松散排列呈海绵状，层次不清，叶绿体少(图 1A;图 1E)。盐碱复合胁迫后，栅栏组织细胞呈短圆柱状，海绵组织细胞变小，叶肉组织细胞排列松散，个别砧木叶片的栅栏组织趋于海绵化，栅栏组织与海绵组织之间没有明确的边界(图 1B;图 1F)。叶片主脉横切面显微结构显示，叶片的主脉是由发达的维管束和机械组织构成，其中维管束由木质部、形成层和韧皮部组成，59 层清晰径向排列的细胞组成木质部，多层小细胞组成韧皮部，木质部占整个维管hensis var.pingyiensis,and the weaker M.sieversii,M.chaenomeles,M.baccata,M.hupehensis.The results ofthis study provide some theoretical basis and reference value fo