Journal of Hebei University(Natural Science Edition) ›› 2021, Vol. 41 ›› Issue (4): 406-411.DOI: 10.3969/j.issn.1000-1565.2021.04.010

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Staining methods on endophytic fungi in Lycium barbarum roots

CHEN Sijie,LI Jin,DU Juan,JIA Baosen,GU Peiwen   

  1. College of Agriculture, Ningxia University, Yinchuan 750021, China
  • Received:2021-03-09 Published:2021-09-03

Abstract: In order to better observe the infection and colonization characteristics of endophytic arbuscular mycorrhizal fungi(AMF)and dark seprate endophytes(DSE)in Lycium barbarum roots, an optimal staining method of endophytic fungi in L. barbarum roots was established. The healthy L. barbarum roots were placed in 100 g/L KOH solution at 90 ℃ for 90 min, then bleached with 30% H2O2 for 5 min, acidified with lactic acid for 5 min, and dyed with vinegar and ink, trypan blue, acid fuchsin, Sudan Ⅳ and aniline blue for 3—5 min, root samples were soaked in decolorant and then examined under micriscope. The results showed that vinegar and Quink ink and trypan blue had the best staining effect. The hyphae and vesicles of AMF and the hyphae and microsclerotia of DSE in the cortical cells of roots can be clearly seen, which can accurately distinguish AMF, DSE and other fungi. However, considering the difficulty and safety of operation, vinegar and Quink ink is more suitable for staining and observation of endophytic fungi in L. barbarum roots because of its simple operation, safety, non-toxicity and good staining effect.

Key words: Lycium barbarum roots, endophytic fungi, arbuscular mycorrhizal fungi, dark seprate endophytes, staining

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