枸杞根系中AMF染色方法优化及其侵染率与土壤因子的关系  

Staining methods on arbuscular mycorrhizal fungi in Lycium barbarum roots and the relationship between colonization rate and soil factors

作　　者：杨沐 季生连 郭寰 段国珍 樊光辉[1,3] 李建领 王占林 YANG Mu;JI Shenglian;GUO Huan;DUAN Guozhen;FAN Guanghui;LI Jianling;WANG Zhanlin(Academy of Agricultural and Forestry Sciences,Qinghai University,Xining 810016,Qinghai,China;College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,Qinghai,China;Qinghai Plateau Key Laboratory of Tree Genetics and Breeding,Xining 810016,Qinghai,China)

机构地区：[1]青海大学农林科学院,青海西宁810016 [2]青海大学农牧学院,青海西宁810016 [3]青海高原林木遗传育种实验室,青海西宁810016

出　　处：《山东大学学报（理学版）》2024年第11期40-50,共11页Journal of Shandong University(Natural Science)

基　　金：青海省科技计划基础研究项目(2021-ZJ-962Q);青海大学科研基金项目(2020-QNY-3)。

摘　　要：了解丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)侵染及定殖情况是深入研究调动土著AMF提高植物抗性的基础。以柴达木盆地9个不同地区8年生大田枸杞为研究对象,基于AMF基本的染色镜检法,在前人研究的基础上,对影响AMF观测的主要因素进行优化研究,并比较不同地区枸杞侵染率及土壤理化性质对其的影响。结果表明,柴达木地区8年生大田枸杞根系AMF侵染率观测效果最佳步骤:首先将FAA固定液固定好的根段置于质量分数10%KOH水溶液中,90℃水浴90 min,后置于沸腾的碱性双氧水(3 mL NaOH+30 mL质量分数10%H_(2)O_(2)溶液+H_(2)O至600 mL)中脱色15~20 min,接着放入质量分数30%H_(2)O_(2)溶液漂白5 min,乳酸酸化5 min,醋酸墨水染色5 min,乳酸甘油90℃下褪色30min。利用该方法可清晰观察到大田多年生枸杞根系中AMF的泡囊和菌丝结构。通过多元统计分析可知柴达木不同地区枸杞根系AMF平均侵染率为41.32%,冗余分析(redundancy analysis,RDA)显示,枸杞根系中的AMF侵染率主要受到土壤pH、土壤中有效磷和速效钾含量等因子的影响,其中土壤pH与AMF侵染率呈显著正相关关系,而有效磷和速效钾含量与侵染率呈显著负相关关系。Understanding the infection and colonization of arbuscular mycorrhizal fungi(AMF)is the basis for mobilizing indigenous AMF to improve plant resistance.Lycium barbarum in nine fields of the Qaidam Basin were the subject.Based on the basic staining method of AMF,previous studies were optimised to investigate the main factors influencing the observation of AMF on the roots of perennial field-grown L.barbarum.The influence of different regions,soil pH,soil physicochemical properties and the colonization rate of AMF on L.barbarum were also compared.The results showed that the optimal observation effect of AMF colonization rate on the roots of Perennial field-grown L.barbarum in the Qaidam Basin was achieved by the following steps.First,the fixed root segments were placed in a 10%KOH 90℃for 90 min,then in the boiling alkaline peroxide(3 mL NaOH+30 mL 10%H_(2)O_(2)+H_(2)O to 600 mL)for 15-20 min,followed by 5 min of H_(2)O_(2)bleaching,5 min of lactic acidification,5 min of acetone ink staining and 30 min of lactic acid glycerol 90℃decolourisation.This method can clearly observe the structures of vesicles and hyphae in the roots of perennial field-grown L.barbarum.The average AMF colonization rate of perennial field-grown L.barbarum in the Qaidam Basin was about 41.32%.RDA analysis showed that the AMF colonization rate in L.barbarum roots was mainly affected by factors such as soil pH,available phosphorous content(xAP),and available potassium content(xAK),among which the soil pH showed a significant positive correlation with the AMF colonization rate,while xAP and xAK showed a significant negative correlation with the colonization rate.

关 键 词：枸杞 丛枝菌根真菌 侵染率 冗余分析 

分 类 号：Q948.12[生物学—植物学] S718.81[农业科学—林学]