Identification and Biological Analysis of Fusarium lateritium Causing Leaf Blight Disease on Coffea canephora

doi:10.3969/j.issn.1000-2561.2022.11.019

Abstract

Abstract:

An unknown disease on Coffea canephora which caused the leaves to become brick-red and withered in a farm of Baisha County, Hainan Province was studied. Two pure cultures 21BS02-1 and 21BS02-2 were isolated. The isolates were inoculated into the leaves and the symptoms were observed. It was found that the symptoms of 21BS02-1 were similar to the field symptoms, and the re-isolate was also consistent with the original pathogen. Therefore, strain 21BS02-1 was determined to be the pathogen. The colony of strain 21BS02-1 was mainly white, felt, dense hyphae, and the color of the center colony was rose. Its mycelia were slender, and some mycelia were septate mycelium. The two ends of macroconidia were slightly curved and shaped like a sickle, with a size of (56.26-175.76)μm × (12.93-19.78)μm, and there were 3-7 partitions. There were few microconidium, which were mainly elliptical and with 0-1 partition. According to the morphological characteristics, the pathogen was preliminarily identified as Fusarium sp. Then through internal transcribed spacer (ITS), β-tubulin and TEF genes to identify the pathogen, BLAST indicated that the homology with Fusarium lateritium of each gene sequence was 99.56% (MN686293), 100% (KJ00154), 99.68% (KF918550), respectively. The three single gene cluster trees based on ITS, β- tubulin, and TEF gene sequence showed that 21BS02-1 belonged to F. lateritium. The determination of biological characteristics showed that the most suitable medium for 21BS02-1 was OMA and CMA medium. The most suitable carbon source was sucrose, and the utilization rate was the highest. Beef extract was the most suitable nitrogen source. The mycelia growth was the fastest under the condition of alternating light and dark for 12 h, and pH value 7-9. This is the first detailed report of F. lateritium infecting C. canephora.

Key words: coffee, leaf blight disease, pathogen identification, Fusarium lateritium

CLC Number:

S435.712

WANG Qian, WU Weihuai, HE Chunping, LIANG Yanqiong, LU Ying, YI Kexian. Identification and Biological Analysis of Fusarium lateritium Causing Leaf Blight Disease on Coffea canephora[J]. Chinese Journal of Tropical Crops, 2022, 43(11): 2345-2355.

Figures/Tables 10

Fig. 1 Disease symptoms and pathogen morphology A, B, C: Symptom in the field; D: Colony on PDA medium (front and back); E: Mycelial morphology of the strain; F: Conidiophore and macroconidia; G: Morphology of the macroconidia.

Fig. 2 Pathogenicity test symptoms and pathogen morphology A, B: Leaf symptoms of inoculated pathogen for 3, 4 d, the left dashed frame was CK, the right side solid box represented the symptoms of inoculated pathogen; C: Symptoms of no stab inoculation for 30 d; D: Symptoms of no stab inoculation for 60 d; E, F: Morphology of the macroconidia and microconidium were reisolated; G: Microconidium; H: Macroconidia.

Fig. 3 Phylogenetic tree based of strain 21BS02-1 based on ITS gene sequence

Fig. 4 Phylogenetic tree based of strain 21BS02-1 based on β-tubulin gene sequence

Fig. 5 Phylogenetic tree based of strain 21BS02-1 based on TEF gene sequence

Fig. 6 Effects of different media on mycelial growth and spore germination of strain 21BS02-1 Different lowercase letters indicate significant difference (P<0.05).

Fig. 7 Effects of different carbon sources on mycelial growth and spore germination of strain 21BS02-1 Different lowercase letters indicate significant difference (P<0.05).

Fig. 8 Effects of different nitrogen sources on mycelial growth and spore germination of strain 21BS02-1 Different lowercase letters indicate significant difference (P<0.05).

Fig. 9 Effects of different light conditions on mycelial growth and spore germination of strain 21BS02-1 Different lowercase letters indicate significant difference (P<0.05).

Fig. 10 Effects of different pH value on mycelial growth and spore germination of strain 21BS02-1 Different lowercase letters indicate significant difference (P<0.05).

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