Role of Brassica rapa SWEET genes in the defense response to Plasmodiophora brassicae

• Published in: Genes & genomics 2024-02, Vol.46 (2), p.253-261
• Main Authors: Choi, Jae-Han, Cho, Eun, Kim, Ji-Woo, Lee, Soo Min, Choi, Gyung Ja, Choi, Su Ryan, Yang, Man Sung, Lim, Yong Pyo, Oh, Man-Ho
• Format: Article
• Language: English
• Subjects: Animal Genetics and Genomics; Biomedical and Life Sciences; Brassica - genetics; Brassica oleracea; Brassica rapa; Brassica rapa - genetics; Clubroot; Disease resistance; Host plants; Human Genetics; Life Sciences; Microbial Genetics and Genomics; Mutants; Pathogenicity; Pathogens; Plant diseases; Plant extracts; Plant Genetics and Genomics; Plasmodiophora brassicae; Plasmodiophorida - genetics; Polymerase chain reaction; Research Article; Ribonucleic acid; RNA; Root nodules; Soil nutrients; T-DNA; Water
• ISSN: 1976-9571; 2092-9293; 2092-9293
• DOI: 10.1007/s13258-023-01486-3

Background Interactions of plants with biotic stress factors including bacteria, fungi, and viruses have been extensively investigated to date. Plasmodiophora brassicae , a protist pathogen, causes clubroot disease in Cruciferae plants. Infection of Chinese cabbage ( Brassica rapa ) plants with P. brassica results in the formation of root galls, which inhibits the roots from absorbing soil nutrients and water. Sugar, the major source of carbon for all living organisms including pathogens and host plants, plays an important role in plant growth and development. Objective To explore the roles of BrSWEET2 , BrSWEET13 , and BrSWEET14 in P. brassicae resistance, Arabidopsis thaliana T-DNA knockout mutants sweet2 , sweet13 , and sweet14 were employed. Methods To isolate total RNA from the collected root nodules, the root tissues washed several times with running water and frozen tissues with liquid nitrogen. Total RNA was extracted using the Spectrum™ Plant Total RNA Kit (SIGMA) and cDNA was synthesized in a 20 μl reaction volume using the ReverTra Ace-α- ® kit (TOYOBO). Real-time PCR was performed in a 10 μl reaction volume containing 1 μl of template DNA, 1 μl of forward primer, 1 μl of reverse primer, 5 μl of 2× iQTM SYBR ® Green Supermix (BioRad), and 2 μl of sterile distilled water. The SWEET genes were genotyped using BioFACT™ 2× TaqBasic PCR Master Mix 2. Results Both sweet2 and sweet14 showed strong resistance to P. brassicae compared with wild-type Arabidopsis and Chinese cabbage plants and sweet13 mutant plants. Pathogenicity assays indicated that the SWEET2 gene plays an important role in clubroot disease resistance in higher plants.