Open AccessHost‐induced post‐transcriptional hairpin RNA ‐mediated gene silencing of vital fungal genes confers efficient resistance against F usarium wilt in banana
Author(s) -
Ghag Siddhesh B.,
Shekhawat Upendra K. S.,
Ganapathi Thumballi R.
Publication year - 2014
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12158
Subject(s) - biology , rna , rna silencing , small hairpin rna , gene , rna interference , fusarium oxysporum , intron , gene silencing , fusarium wilt , gene expression , transcription (linguistics) , microbiology and biotechnology , genetics , linguistics , philosophy
Summary Fusarium wilt, caused by F usarium oxysporum f. sp. cubense ( F oc), is among the most destructive diseases of banana ( M usa spp.). Because no credible control measures are available, development of resistant cultivars through genetic engineering is the only option. We investigated whether intron hairpin RNA (ihp RNA )‐mediated expression of small interfering RNA s (si RNA s) targeted against vital fungal genes ( velvet and Fusarium transcription factor 1 ) in transgenic banana could achieve effective resistance against F oc. Partial sequences of these two genes were assembled as ihp RNA s in suitable binary vectors (ihp RNA ‐ VEL and ihp RNA ‐ FTF 1) and transformed into embryogenic cell suspensions of banana cv. R asthali by A grobacterium ‐mediated genetic transformation. Eleven transformed lines derived from ihp RNA ‐ VEL and twelve lines derived from ihp RNA ‐ FTF 1 were found to be free of external and internal symptoms of F oc after 6‐week‐long greenhouse bioassays. The five selected transgenic lines for each construct continued to resist F oc at 8 months postinoculation. Presence of specific si RNA s derived from the two ihp RNA s in transgenic banana plants was confirmed by Northern blotting and I llumina sequencing of small RNA s derived from the transgenic banana plants. The present study represents an important effort in proving that host‐induced post‐transcriptional ihp RNA ‐mediated gene silencing of vital fungal genes can confer efficient resistance against debilitating pathogens in crop plants.