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Genetic Determinism of Sensitivity to Corynespora cassiicola Exudates in Rubber Tree (Hevea brasiliensis)

机译:橡胶树(巴西橡胶树)中对棒状杆菌(Corynespora cassiicola)渗出液敏感性的遗传确定性

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摘要

An indirect phenotyping method was developed in order to estimate the susceptibility of rubber tree clonal varieties to Corynespora Leaf Fall (CLF) disease caused by the ascomycete Corynespora cassiicola. This method consists in quantifying the impact of fungal exudates on detached leaves by measuring the induced electrolyte leakage (EL%). The tested exudates were either crude culture filtrates from diverse C. cassiicola isolates or the purified cassiicolin (Cas1), a small secreted effector protein produced by the aggressive isolate CCP. The test was found to be quantitative, with the EL% response proportional to toxin concentration. For eight clones tested with two aggressive isolates, the EL% response to the filtrates positively correlated to the response induced by conidial inoculation. The toxicity test applied to 18 clones using 13 toxinic treatments evidenced an important variability among clones and treatments, with a significant additional clone x treatment interaction effect. A genetic linkage map was built using 306 microsatellite markers, from the F1 population of the PB260 x RRIM600 family. Phenotyping of the population for sensitivity to the purified Cas1 effector and to culture filtrates from seven C. cassiicola isolates revealed a polygenic determinism, with six QTL detected on five chromosomes and percentages of explained phenotypic variance varying from 11 to 17%. Two common QTL were identified for the CCP filtrate and the purified cassiicolin, suggesting that Cas1 may be the main effector of CCP filtrate toxicity. The CCP filtrate clearly contrasted with all other filtrates. The toxicity test based on Electrolyte Leakage Measurement offers the opportunity to assess the sensitivity of rubber genotypes to C. cassiicola exudates or purified effectors for genetic investigations and early selection, without risk of spreading the fungus in plantations. However, the power of this test for predicting field susceptibility of rubber clones to CLF will have to be further investigated.
机译:为了估计橡胶树无性系变种对由子囊虫Corynespora cassiicola引起的Corynespora Leaf Fall(CLF)疾病的敏感性,开发了一种间接表型方法。该方法包括通过测量诱导的电解质泄漏(EL%)来量化真菌渗出物对离体叶片的影响。所测试的分泌物是来自各种C. cassiicola分离株的粗培养滤液或纯化的cassiicolin(Cas1)(由侵略性分离株CCP产生的少量分泌的效应蛋白)。发现该测试是定量的,EL%响应与毒素浓度成正比。对于用两个攻击性分离物测试的八个克隆,对滤液的EL%响应与分生孢子接种诱导的响应呈正相关。使用13种毒素处理方法对18个克隆进行的毒性测试证明,克隆和处理方法之间存在重要的变异性,并且具有明显的附加x相互作用。使用来自PB260 x RRIM600家族F1群体的306个微卫星标记构建了遗传连锁图谱。对纯化的Cas1效应子和培养的7个C. cassicola菌株的滤液的敏感性进行群体表型分析,揭示了一种多基因确定性,在5条染色体上检测到6个QTL,解释表型变异的百分比从11%到17%不等。对于CCP滤液和纯化的卡西林可鉴定出两个常见的QTL,这表明Cas1可能是CCP滤液毒性的主要影响因子。 CCP滤液与所有其他滤液明显不同。基于电解质泄漏测量的毒性测试提供了机会,可以评估橡胶基因型对C. cassiicola渗出液或纯化的效应子的敏感性,以进行遗传研究和早期选择,而没有将真菌传播到人工林中的风险。但是,该测试对于预测橡胶克隆对CLF的现场敏感性的能力将有待进一步研究。

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