Physiological and biochemical characteristics of boscalid resistant isolates of Sclerotinia sclerotiorum from asparagus lettuce

doi:10.1016/j.jia.2023.09.024

摘要/Abstract

摘要：

由核盘菌（Sclerotinia sclerotiorum）引起的莴苣菌核病是我国常见的病害。由于缺乏可商业化的抗性品种，目前主要依赖杀菌剂进行防控。啶酰菌胺（boscalid, 简称Bos），隶属于琥珀酸脱氢酶抑制剂类（SDHI）杀菌剂，在我国用于核盘菌防治已有十多年，部分地区出现该药剂对莴苣菌核病的防效下降现象，但核盘菌的抗药性状况尚不清楚。为此，本实验室于2019年从江苏六个市的莴苣病样中共分离了172株核盘菌。采用鉴别剂量（5 μg mL^-1的浓度）法，进行抗药性菌株鉴定，发现对该药剂的低抗（Bos^LR）群体占所检测菌株总数的76.74%。进一步研究表明：Bos^LR菌株的生物适合度不低于啶酰菌胺敏感（Bos^S）菌株；大多数Bos^LR菌株的草酸积累和胞外多糖（EPS）含量较Bos^S菌株均增加，但膜透性降低，对供试的胁迫因子耐受性差异显著；离体防效测定也验证了田间确实存在Bos^L^R的抗药性亚群体；田间存在3种对啶酰菌胺的抗性基因型（①SDHB^A11V，②SDHC^Q38R，③SDHB^A11V+SDHC^Q38R），该抗性可能由药剂和靶标间的亲和力下降导致；但Bos^LR菌株对噻呋酰胺、氟唑菌酰羟胺、氟啶胺和戊唑醇间均无交互抗性。本研究首次报道了我国莴苣核盘菌对啶酰菌胺存在田间抗性，这对该病的抗药性治理和精准防控具有指导意义。

Abstract:

Laboratory mutants of Sclerotinia sclerotiorum (Lib) de Bary, resistant to boscalid, have been extensively characterized. However, the resistance situation in the lettuce field remains largely elusive. In this study, among the 172 S. sclerotiorum isolates collected from asparagus lettuce field in Jiangsu Province, China, 132 isolates (76.74%) exhibited low-level resistance to boscalid (Bos^LR), with a discriminatory dose of 5 μg mL^–1. In comparison to the boscalid-sensitive (Bos^S) isolates, most Bos^LR isolates demonstrated a slightly superior biological fitness, as evidenced by data on mycelial growth, sclerotium production and pathogenicity. Moreover, most Bos^LR isolates showed comparable levels of oxalic acid (OA) accumulation, increased exopolysaccharide (EPS) content and reduced membrane permeability when compared to the Bos^S isolates. Nevertheless, their responses to distinct stress factors diverged significantly. Furthermore, the effectiveness of boscalid in controlling Bos^LR isolates on radish was diminished compared to its efficacy on Bos^S isolates. Genetic mutations were identified in the SDH genes of Bos^LRisolates, revealing the existence of three resistant genotypes: I (A11V at SDHB, SDHB^A11V), II (Q38R at SDHC, SDHC^Q38R) and III (SDHB^A11V+SDHC^Q38R). Importantly, no cross-resistance was observed between boscalid and other fungicides such as thifluzamide, pydiflumetofen, fluazinam, or tebuconazole. Our molecular docking analysis indicated that the docking total score (DTS) of the type I resistant isolates (1.3993) was lower than that of the sensitive isolates (1.7499), implying a reduced affinity between SDHB and boscalid as a potential mechanism underlying the boscalid resistance in S. sclerotiorum. These findings contribute to an enhanced comprehension of boscalid’s mode of action and furnish valuable insights into the management of boscalid resistance.

Key words: Sclerotinia sclerotiorum , Boscalid , Asparagus lettuce , SDHB^A11V , SDHC^Q38R

SHI Dong-ya, LI Feng-jie, ZHANG Zhi-hui, XU Qiao-nan, CAO Ying-ying, Jane Ifunanya MBADIANYA, LI Xin, WANG Jin, CHEN Chang-jun. [J]. Journal of Integrative Agriculture, 2023, 22(12): 3694-3708.

SHI Dong-ya, LI Feng-jie, ZHANG Zhi-hui, XU Qiao-nan, CAO Ying-ying, Jane Ifunanya MBADIANYA, LI Xin, WANG Jin, CHEN Chang-jun. Physiological and biochemical characteristics of boscalid resistant isolates of Sclerotinia sclerotiorum from asparagus lettuce[J]. Journal of Integrative Agriculture, 2023, 22(12): 3694-3708.

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