Acta Agriculturae Boreali-Sinica ›› 2022, Vol. 37 ›› Issue (2): 183-191. doi: 10.7668/hbnxb.20192811

Special Issue: Pear Hot Article

• Resources & Environment·Plant Protection • Previous Articles     Next Articles

Effect and Mechanism of the Disease Resistance to Pear Black Spot Induced by Salicylic Acid in Yali

WEI Yarui1, ZHAO Shuliang2, CHENG Xiaohua1, YAN Qi1, LIU Na1, ZHANG Yuxing1   

  1. 1.College of Horticulture,Hebei Agricultural University,Pear Engineering Technology Research Center of Hebei Province,Baoding 071000,China
    2.School of Landscape and Ecological Engineering,Hebei University of Engineering,Handan 056038,China
  • Received:2021-10-20 Published:2022-04-28

水杨酸诱导鸭梨抗黑斑病效果及机理的探讨

魏亚蕊1, 赵曙良2, 成晓华1, 闫琦1, 刘娜1, 张玉星1   

  1. 1.河北农业大学 园艺学院,河北省梨工程技术研究中心,河北 保定 071000
    2.河北工程大学 园林与生态工程学院,河北 邯郸 056038
  • 作者简介:
    作者简介:魏亚蕊(1986—),女,河北石家庄人,博士,主要从事果树结实生理与分子生物学研究。
  • 基金资助:
    科技研发平台与新型研发机构建设专项(20567676H)

Abstract:

In order to reveal the effect and molecular mechanism of salicylic acid-induced resistance to black spot disease of Yali,the leaves and fruits of Yali were used as experimental material which were collected from the experimental orchard of Hebei Agricultural University.Tissue isolation method was used to isolate and purify the pathogen of black spot disease and detected its pathogenicity.Morphological observation and multi-genes of ITS,HIS,RPB2 and ACT were used to identify the pathogen of black spot disease.The detached leaves of Yali were inoculated with conidia suspension of Alternaria alternata by acupuncture for 0,6,12,24,48,72,96 and 120 h,respectively.Real-time RT-PCR was used to analyse the expression of related genes which involved in salicylic acid signaling,and high-performance liquid chromatography (HPLC) was used to detect the content of endogenous SA at 0 and 72 h,respectively.Exogenous SA treatment with different concentrations (0,0.002,0.02,0.2,2.0,10.0,20.0 mmol/L) were used to identify the effect to the mycelial growth of A.alternata;Exogenous SA treated Yali fruit with different concentrations (0,0.02,0.2,2.0 mmol/L),respectively,and inoculated with A.alternata to detect the effect of disease resistance.The results showed that the pathogen of Yali black spot disease was A.alternata. Detached leaves of Yali were inoculated with A.alternata for 72 h,the free state SA content increased from 0 mg/g to 0.02 mg/g and the binding state SA content increased from 0.47 mg/g to 1.55 mg/g,and for 96 h compared with 0 h,the expression of Pbrgene12425,Pbrgene6286, increased 5.48,4.66 times,respectively,the expression of Pbrgene8895 and Pbrgene43605 increased 7.90 and 10.0 times for 120 h.Exogenous 0.2 mmol/L SA significantly increased the resistance of Yali fruit to black spot disease.In the process of Yali resistance to black spot disease,the free state SA content of Yali leaves significantly increased,and the related genes in SA signaling were induced to up-expressed,exogenous 0.2 mmol/L SA significantly increased the resistance of Yali to black spot disease.

Key words: Yali, Black spot, Alternaria alternata, Salicylic acid, Tissue isolation method, qRT-PCR

摘要:

为了揭示水杨酸诱导鸭梨抗黑斑病的作用效果及机理,以河北农业大学实验农场梨园鸭梨叶片和果实为试验材料,通过组织分离法分离、纯化梨黑斑病病原菌并对其进行致病性检测,利用形态观察及真菌 ITSHISRPB2ACT多基因综合鉴定法明确鸭梨黑斑病致病菌的类型;鸭梨离体叶片通过针刺接种黑斑病菌分生孢子悬浮液侵染0,6,12,24,48,72,96,120 h,通过实时荧光定量PCR检测水杨酸信号通路中相关基因的表达量变化,通过高效液相色谱(HPLC)检测0,72 h内源水杨酸含量的变化;利用不同浓度水杨酸(0,0.002,0.02,0.2,2.0,10.0,20.0 mmol/L)测定其对黑斑病菌菌丝生长的影响;采用外源0,0.02,0.2,2.0 mmol/L水杨酸处理鸭梨果实,接种黑斑病菌并观察其抗病效果。结果表明,河北农业大学实验农场梨园中鸭梨黑斑病病原菌为链格孢属链格孢菌,鸭梨离体叶片接种黑斑病菌后,72 h游离态水杨酸含量由0 mg/g增至0.02 mg/g,结合态水杨酸含量由0.47 mg/g增至1.55 mg/g,SA信号通路中相关基因Pbrgene12425Pbrgene6286接种96 h与0 h相比表达量分别上调5.48,4.66倍,Pbrgene8895Pbrgene43605接种120 h表达量上调7.90,10.0倍,外源施加0.2 mmol/L水杨酸显著提高鸭梨果实对黑斑病的抗性。在鸭梨抗黑斑病过程中,鸭梨离体叶片游离态水杨酸含量显著增加,水杨酸信号通路中相关基因被诱导上调表达,外源施加0.2 mmol/L水杨酸可提高鸭梨果实对黑斑病的抗性。

关键词: 鸭梨, 黑斑病, 链格孢菌, 水杨酸, 组织分离法, 荧光定量PCR