西红花茎腐病致病真菌的分离与鉴定

  目的  真菌性病害是西红花Crocus sativus的主要病害。分离鉴定西红花茎腐病致病菌可为西红花茎腐病科学防控及特效杀菌剂开发提供科学依据。  方法  以‘番红花1号’C. sativus ‘Saffron No. 1’为供试材料，采用组织分离法从感病球茎中分离获得致病真菌。通过PCR扩增、测序与进化树构建对真菌保守内部转录间隔区序列(ITS)和RNA聚合酶 Ⅱ 亚家族(RPB2)基因序列进行同源性比对与进化分析。对感病球茎进行致病菌再分离并回接到植株，验证真菌在种球种植模式下的致病特性。  结果  所分离获得的球茎致病真菌与黑曲霉Aspergillus niger的形态特征高度相似；候选致病真菌的ITS和RPB2保守序列与其他植物来源的黑曲霉同一DNA序列同源性高达99.6%，与黑曲霉处在同一进化分支上；基质种植模式下，接种黑曲霉菌液极显著(P＜0.01)增加了西红花球茎的发病率。  结论  黑曲霉是引起西红花种球腐烂的真菌病害之一。图7参32

Isolation and identification of pathogenic fungi of stem rot in Crocus sativus

  Objective  This study, with an investigation into the identification and isolation of the rot disease pathogen of Crocus sativus, the main disease of which is fungal disease, is intended to provide scientific basis for the prevention and control of saffron stem rot and the development of special fungicides.   Method  First, the pathogenic fungi were isolated from infected corms by tissue separation method using ‘Saffron No. 1’ as the test material. Then an analysis was made of the sequences of conserved internal transcribed spacer (ITS) and RNA polymerase Ⅱ subfamily (RPB2) by PCR amplification, sequencing and phylogenetic tree construction. At last, pathogenic bacteria were isolated from infected corms and grafted back to plants to verify the pathogenic characteristics of fungi under the planting mode of saffron.   Result  The morphological characteristics of the corm-pathogenic fungi isolated were highly similar to those of Aspergillus niger. The conserved sequences of ITS and RPB2 of the candidate pathogenic fungi had 99.6% homology to the same DNA sequence of A. niger from other plants with the same evolutionary branch. In the soil planting mode, inoculation with A. niger solution significantly increased the incidence of corm (P＜0.01).   Conclusion  A. niger is one of the fungal diseases that cause the rot of saffron bulbs. Ch, 7 fig. 32 ref.]