Sains Malaysiana 47(10)(2018): 2291–2300

http://dx.doi.org/10.17576/jsm-2018-4710-05

 

The Effect of Pathogenesis-Related 10 (Pr-10) Gene on the Progression of Fusarium Wilt in Musa acuminata cv. Berangan

(Kesan Gen Berkaitan Patogenesis 10 (Pr-10) pada Perkembangan Fusarium Layu dalam Musa acuminata cv. Berangan)

 

NADIYA AKMAL BAHARUM^1,2, ROFINA YASMIN OTHMAN^1,3, YUSMIN MOHD-YUSUF³, BOON CHIN TAN³, KAMILATULHUSNA ZAID1^I,3 & NORZULAANI KHALID^1,3*

 

¹Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

 

²Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia

 

³Centre for Research in Biotechnology for Agriculture (CEBAR), University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

 

⁴Centre for Foundation Studies in Science, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

 

Received: 15 December 2017/Accepted: 19 June 2018

 

 

ABSTRACT

PR-10 is a member of pathogenesis-related (PR) genes elicited by the plant’s defense mechanism during pathogen attack. Elevated expression of PR-10 upon different pathogen invasions has been observed in many plant species suggesting its role as an anti-bacterial, anti-viral and anti-fungal gene. However, the effect of PR-10 in mitigating the infection of Fusarium oxysporum f. sp. cubense (Foc), the causal agent of Fusarium wilt in banana has not been reported. In this study, the coding sequences of PR-10 gene isolated from Foc resistant Musa acuminata ssp. malaccensis (MaPR-10) were integrated into a local Foc susceptible commercial banana cultivar, ‘Berangan’ via co-cultivation of embryogenic cell suspension and Agrobacterium tumefaciens. Out of 17 putative transgenic lines established, 11 of them positively harbored MaPR-10. Among these, Line-19 plantlets showed the most rapid in-vitro propagation and successfully over-expressed the transgene. Following a nursery challenge experiment with a virulent Foc race 4 (CI HIR) isolate, about 30% of Line-19 plants showed a one-week delay in disease progression when compared to the untransformed controls. From the final evaluation performed in the 5th week-post-inoculation, the leaf symptoms index (LSI) and rhizome discoloration index (RDI) of Line-19 was 3.4 and 6.1, respectively, indicating the disease had progressed. The findings of this study enrich the current existing knowledge on the roles of PR-10 in combating fungal disease in plants.

 

Keywords: Fusarium oxysporum f. sp. cubense; Fusarium wilt; pathogenesis-related protein; PR-10; transgenic banana

 

ABSTRAK

PR-10 merupakan antara gen berkaitan patogenan (PR) yang dihasilkan oleh sistem pertahanan tumbuhan semasa serangan patogen. Peningkatan ekspresi gen PR-10 ketika ancaman beberapa jenis patogen yang berbeza telah diperhatikan di dalam banyak spesies tumbuhan menunjukkan peranan gen ini sebagai agen anti-bakteria, anti-virus dan anti-kulat. Namun begitu, peranan PR-10 dalam mengurangkan kesan jangkitan Fusarium oxysporum f. sp. cubense (Foc), patogen penyebab penyakit layu pada pokok pisang masih belum pernah dilaporkan. Dalam kajian ini, jujukan pengekodan PR-10 daripada Musa acuminata ssp. malaccensis (MaPR-10) telah diintegrasikan ke dalam genom satu kultivar pisang komersial tempatan, ‘Berangan’ melalui melalui kaedah penanaman antara Agrobacterium tumefaciens dan ampaian sel embriogenik. Daripada 17 jaluran pokok transgenik yang terhasil, 11 daripadanya disahkan positif mengandungi gen MaPR-10. Antara kesemua jaluran yang terhasil, Jaluran 19 menunjukkan kadar propagasi in-vitro paling cepat dan menunjukkan ekspresi gen yang lebih tinggi berbanding pokok kawalan yang tidak ditransform. Dalam penilaian tindak balas Jaluran 19 terhadap jangkitan Foc ‘race’ 4 (C1 HIR), 30% daripada anak pokok menunjukkan penularan penyakit seminggu lebih lambat berbanding pokok kawalan yang tidak ditransform. Melalui penilaian terakhir pada minggu ke-5 selepas inokulasi, skor indeks sistem daun (LSI) dan indeks penyahwarnaan rizom (RDI) bagi Jaluran 19 masing-masing ialah 3.4 dan 6.1, menunjukkan bahawa jangkitan Foc telah merebak. Keputusan kajian uji kaji ini berupaya meningkatkan maklumat sedia ada berkaitan peranan PR-10 dalam menentang penyakit kulat di dalam tumbuhan.

 

Kata kunci: Fusarium oxysporum f. sp. cubense; penyakit layu Fusarium; pisang transgenik; protein berkaitan patogenan; PR-10

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*Corresponding author; email: lani@um.edu.my