Monitoring peroxisome dynamics using enhanced green fluorescent protein labeling in Alternaria alternata

Ziqi Lu; Jian Guo; Qiang Li; Yatao Han; Zhen Zhang; Zhongna Hao; Yanli Wang; Guochang Sun; Jiaoyu Wang; Ling Li

doi:10.3389/fmicb.2022.1017352

Monitoring peroxisome dynamics using enhanced green fluorescent protein labeling in Alternaria alternata

Front Microbiol. 2022 Oct 25:13:1017352. doi: 10.3389/fmicb.2022.1017352. eCollection 2022.

Authors

Ziqi Lu^{1

2}, Jian Guo³, Qiang Li¹, Yatao Han¹, Zhen Zhang², Zhongna Hao², Yanli Wang², Guochang Sun², Jiaoyu Wang², Ling Li¹

Affiliations

¹ The Key Laboratory for Quality Improvement of Agricultural Products of Zhejiang Province, College of Advanced Agricultural Sciences, Zhejiang Agriculture and Forestry University, Hangzhou, China.
² State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou, China.
³ College of Food and Health (College of Modern Food Industry), Zhejiang Agriculture and Forestry University, Hangzhou, China.

Abstract

Brown leaf spot on tobacco is a serious fungal disease caused by Alternaria alternata. Peroxisomes are organelles playing an important role in the development and infection of plant pathogenic fungi. But, until now, there is no report on the peroxisome dynamics during the conidia germination of A. alternata. To evaluate the roles of peroxisome in the development of the fungus, in the present work, an enhanced green fluorescent protein (eGFP) cassette tagged with peroxisome targeting signal 2 (PTS2) was integrated into A. alternata to label the organelles, and an eGFP cassette carrying a nuclear located signal (NLS) was performed parallelly. The transformants containing the fusions emitted fluorescence in punctate patterns. The fluorescence of eGFP-PTS2 was distributed exactly in the peroxisomes while those of eGFP-NLS were located in the nucleus. Typical AaGB transformants were selected to be investigated for the peroxisome dynamics. The results showed that during spore germination, the number of peroxisomes in the spores decreased gradually, but increased in the germ tubes. In addition, when the transformants were cultured on lipid media, the numbers of peroxisomes increased significantly, and in a larger portion, present in striped shapes. These findings give some clues for understanding the peroxisomal functions in the development of A. alternata.

Keywords: Alternaria alternata; dynamic analysis; eGFP-PTS2; lipid; peroxisome.

Grants and funding

We gratefully acknowledge the financial support provided by the National Natural Science Foundation of China (Grant Nos. 31900126 and 31701723), the Zhejiang Key Research and Development Program (Grant No. 2021C02010), the Fundamental Research Funds for the Provincial Universities of Zhejiang (Grant No. 2020YQ002), the Zhejiang Provincial Natural Science Foundation of China under (Grant No. LZ20C140001), the State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Argo-Products (Grant No. 2021DG700024-KF202110), and the Program of Innovative Entrepreneurship Training for Undergraduate of China (Grant No. 202110341006).