Identification of the Neopestalotiopsis musae Pathogen of Banana Leaf Spot Disease

doi:10.16420/j.issn.0513-353x.2021-1115

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (2): 410-420.doi: 10.16420/j.issn.0513-353x.2021-1115

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Identification of the Neopestalotiopsis musae Pathogen of Banana Leaf Spot Disease

LIANG Jiali¹, WU Qisong¹, CHEN Guangquan², ZHANG Rong¹^,³, XU Chunxiang¹, FENG Shujie¹^,³^,^*()

1. College of Horticulture，South China Agricultural University，Guangzhou 510642，China
2. Maoming Agricultural Technology Extension Center，Maoming，Guangdong 525000，China
3. Guangdong Province Key Laboratory of Microbial Signals and Disease Control，Integrative Microbiology Research Centre，South China Agricultural University，Guangzhou 510642，China

Received:2022-08-03 Revised:2022-10-14 Online:2023-02-25 Published:2023-03-06
Contact: *（E-mail：sjief@scau.edu.cn）

Abstract

Abstract:

A new emerging banana leaf spot disease was found during a disease survey in the banana-producing area of Maoming City，Guangdong Province. The pathogen was isolated and tested for pathogenicity according to Koch's postulate，and the pathogen was identified based on morphological characteristics and sequence analyses of ITS，β-tubulin and tef1 genes. The results showed that strain MM3-2z9 could infect banana leaves and induce similar symptoms as in the field. The colony of strain MM3-2z9 on the PDA medium was white，producing obvious concentric ring patterns. The conidia were fusoid with 4-septate，three median cells versicoloured，apical cell hyaline，pyramidal，with two to four filiform apical appendages（mostly three），and basal cell with appendage single was centric and hyaline. Sequence homology analysis indicated that the ITS，β-tubulin and tef1 sequences of three single-conidial strains of MM3-2z9 were more than 98% identical to the corresponding sequences of Neopestalotiopsis musae stains. The phylogenetic tree constructed by the three fragments showed that strain MM3-2z9 was most closely related to N. musae. Therefore，strain MM3-2z9 was identified as N. musae based on morphological analysis and phylogenetic tree analysis. In addition to damaging banana（Musa AAA Cavendish）leaves，the pathogen could also quickly infect the leaves of M. × paradisiaca ABB and Musa ABB Pisang Awak‘Fenza 1’.

Key words: banana, leaf spot disease, pathogen identification, Neopestalotiopsis musae

CLC Number:

S668.1

LIANG Jiali, WU Qisong, CHEN Guangquan, ZHANG Rong, XU Chunxiang, FENG Shujie. Identification of the Neopestalotiopsis musae Pathogen of Banana Leaf Spot Disease[J]. Acta Horticulturae Sinica, 2023, 50(2): 410-420.

Figures/Tables 8

Table1 Primer information used in this study

基因 Gene	引物名称 Primer name	引物序列（5′-3′） Primer sequence	退火温度/℃ Annealing temperature
ITS	ITS4	TCCTCCGCTTATTGATATGC GC	57
ITS	ITS5	GGAAGTAAAAGTCGTAACAAGG
β-tubulin	BT2A	GGTAACCAAATCGGTGCTTTC	58
β-tubulin	BT2B	ACCCTCAGTGTAGTGACCCTTGGC
Tef1	EF1-688F	CGGTCACTTGATCTACAAGTGC	59
Tef1	EF1-986R	TACTTGAAGGAACCCTTACC

Fig. 1 Early（A，B）and late（C）symptoms of new leaf spot of banana in the field

Fig. 2 Symptoms of banana leaves inoculated with three monospora strains of MM3-2z9（11 days）

Fig. 3 Pathogenic symptoms of MM3-2z9 in Musa × paradisiaca ABB and Musa ABB Pisang Awak inoculated for seven days

Fig. 4 Morphological characteristics of pathogenic bacteria A，B：Positive and negative sides of colony on PDA medium；C：Conidia；D：Sporulation cells and primary conidia；E：Sporulation cell；F-H：Conidia.

Fig. 5 Electrophoretic detection of ITS（ITS4/ITS5）amplified bands of three monospore strains（A，B，C）of MM3-2z9

Fig. 6 Molecular phylogenetic tree constructed by NJ method based on tandem of ITS sequence Branch node displays the branch node self-expansion value of NJ tree. The same below.

Fig. 7 Molecular phylogenetic tree constructed by NJ method based on tandem of ITS，β-butulin，and tef1 sequences

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Identification of the Neopestalotiopsis musae Pathogen of Banana Leaf Spot Disease

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