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Yip M.-K.,Academia Sinica, Taiwan | Lee S.-W.,Taiwan Banana Research Institute | Su K.-C.,Academia Sinica, Taiwan | Lin Y.-H.,Academia Sinica, Taiwan | And 2 more authors.
Plant Biotechnology Reports | Year: 2011

This study describes an efficient protocol for Agrobacterium tumefaciens-mediated transformation of two subgroups of genotype AAA bananas (Musa acuminata cv. Pei Chiao and Musa acuminata cv. Gros Michel). Instead of using suspension cells, cauliflower-like bud clumps, also known as multiple bud clumps (MBC), were induced from sucker buds on MS medium containing N6-Benzylaminopurine (BA), Thidiazuron (TDZ), and Paclobutrazol (PP333). Bud slices were co-cultivated with A. tumefaciens C58C1 or EHA105 that carry a plasmid containing Arabidopsis root-type ferredoxin gene (Atfd3) and a plant ferredoxin-like protein (pflp) gene, respectively. These two strains showed differences in transformation efficiency. The EHA105 strain was more sensitive in Pei Chiao, 51. 3% bud slices were pflp-transformed, and 12. 6% slices were Atfd3-transformed. Gros Michel was susceptible to C58C1 and the transformation efficiency is 4. 4% for pflp and 13. 1% for Atfd3. Additionally, gene integration of the putative pflp was confirmed by Southern blot. Resulting from the pathogen inoculation assay, we found that the pflp transgenic banana exhibited resistance to Fusarium oxysporum f. sp. cubense tropical race 4. This protocol is highly advantageous to banana cultivars that have difficulties in setting up suspension cultures for the purpose of quality improvement through genetic transformation. In addition, this protocol would save at least 6 months in obtaining explants for transformation and reduce labor for weekly subculture in embryogenic cell suspension culture systems. © 2011 Korean Society for Plant Biotechnology and Springer.


Li M.H.,South China Agricultural University | Yang B.,University of Hawaii at Manoa | Leng Y.,North Dakota State University | Chao C.P.,Taiwan Banana Research Institute | And 4 more authors.
Canadian Journal of Plant Pathology | Year: 2011

Fusarium wilt of banana (Musa spp.), caused by Fusarium oxysporum f. sp. cubense (Foc), is one of the most important banana diseases. Three races (1, 2, 4) of Foc have been identified worldwide, but race 4 (especially tropical race 4, TR4) is the most destructive pathogen and has significant impacts on banana production in Taiwan and Southern China. To characterize the genetic relationship of Foc in these two regions, 55 isolates of Foc, including 18 from Taiwan and 37 from Hainan and Guangdong Provinces, were subjected to pathogenicity tests, race 4-specific polymerase chain reaction (PCR) diagnosis, and amplified fragment length polymorphism (AFLP) analysis. One race 4 isolate from Australia, one race 1 isolate from Philippines, and 10 non-banana F. oxysporum isolates were also included in the study for comparison. Pathogenicity tests indicated that 16 and 21 isolates of Foc from Taiwan and Southern China, respectively, were pathogenic on Cavendish banana and classified as race 4, and the remaining isolates were identified as race 1. PCR amplification with race 4 and tropical race 4-specific primers confirmed that all of the race 4 isolates from Taiwan and Southern China were TR4. AFLP analysis with eight EcoRI-MseI primer combinations clustered the 67 F. oxysporum isolates analyzed into two major groups, one containing all race 4 isolates and the other consisting of race 1 isolates along with the non-banana F. oxysporum isolates. No significant divergence was observed among TR4 isolates from Southern China and Taiwan, suggesting that the TR4 isolates from Southern China might share the same lineage as those from Taiwan. © 2011 The Canadian Phytopathological Society.


Molina A.B.,International Rice Research Institute | Sinohin V.O.,International Rice Research Institute | Fabregar E.G.,Lapanday Foods Corporation | Ramillete E.B.,Lapanday Foods Corporation | And 2 more authors.
Acta Horticulturae | Year: 2016

Recent epidemics of Fusarium wilt, caused by the virulent strain tropical race 4 (TR4) of Fusarium oxysporum f. sp. cubense (Foc), pose a serious threat to the banana industry. Four Cavendish (AAA genome) somaclonal variants selections from Taiwan (GCTCV-105, GCTCV-119, GCTCV-218 and GCTCV-219) and three important Philippine local cultivars [Latundan (AAB genome, Silk subgroup), Lakatan (AAA genome, Lakatan subgroup) and Saba (ABB genome, Saba subgroup)], were compared to the commercial Cavendish Grand Naine (AAA genome, Cavendish subgroup) in a heavily Foc-infested soil in the southern Philippines. Experimental plots were planted with 10 tissue culture-derived seedlings, replicated 10 times, and arranged in a completely randomized design. Disease incidence was assessed weekly by monitoring early symptoms of yellowing of older leaves and/or other symptoms such as pseudostem splitting. Onset of disease symptoms was recorded. The infected plants were allowed to develop severe typical symptoms of Fusarium wilt (leaves showing clear yellowing, and marginal necroses appear on older leaves) and then checked for vascular necroses as confirmation. The commercially-grown cultivars Grand Naine and Lakatan showed susceptibility with a disease incidence of 64 and 76%, respectively, in the primary crop; and 79 and 92%, respectively in the ratoon crop. Disease incidence in the GCTCV cultivars ranged from 0-6% in the primary crop and 0-8% in the ratoon crop. Saba showed 0% incidence in the primary as well as in the ratoon crop. Foc VCG 01213/16, the Foc strain associated with TR4, was confirmed in the infected plants using a Polymerase Chain Reaction (PCR)-based diagnostic tool. Results confirm the stability of resistant somaclones to Foc TR4, even in agroecosystems differing from those where these were first developed. They also show that selecting favorable somaclonal variants is a feasible approach in cultivar improvement, and they have a potential role in integrated Foc TR4 management. The study identifies the TR4-resistant cultivar Saba, showing that TR4 does not severely affect all cultivars.


Lin Y.-H.,National Chung Hsing University | Su C.-C.,Taiwan Banana Research Institute | Chao C.-P.,Taiwan Banana Research Institute | Chen C.-Y.,National Chung Hsing University | And 3 more authors.
European Journal of Plant Pathology | Year: 2013

The Fusarium genus causes devastating plant diseases worldwide, in which Fusarium oxysporum is the most serious crop pathogen. Disease monitoring is the basis of integrated pest management of any disease. The lack of rapid, accurate, and reliable device to detect and identify plant pathogens is one of the main limitations in integrated disease management. This study describes an efficient and quantifiable diagnosis method for the specific detection of F. oxysporum f. sp. cubense (Foc) race 4 in field-infected banana. With the optimized PCR parameters using the SCAR (sequence characterized amplified region) primers FocSc-1/FocSc-2 and a real-time PCR strategy, the developed method showed high reproducibility and was very sensitive to detect extremely low quantities of Foc genomic DNA (gDNA). We also found that Foc gDNA in severely symptomatic banana pseudostems and leaves were 6946-fold and 26. 69-fold more than in those of mild-symptomatic banana, respectively. © 2012 KNPV.


Chang C.M.,Taiwan Banana Research Institute | Chao C.P.,Taiwan Banana Research Institute | Huang S.N.,Taiwan Banana Research Institute | Chiang S.C.,Taiwan Banana Research Institute
Acta Horticulturae | Year: 2011

Organic banana farming is an alternative to conventional banana production in Chinese Taipei. Results from a 10-year study on organic banana production at the organic farm of the Taiwan Banana Research Institute (TBRI) indicate that average incidence of Fusarium wilt for four Cavendish cultivars was 25.7% for the organic farming system, as compared to 66.9% for the conventional farming system. The lower incidence of Fusarium wilt in the organic system was considered to be associated with a positive change in the properties of the soil. Horticultural parameters at shooting did not show significant differences between organic and conventional banana, except for a lower number of healthy leaves in the organic banana. No differences in the post-harvest quality could be found between organic and conventional banana. A smaller bunch weight was recorded for the organic banana; however, more harvested bunches resulted in a higher overall yield in the organic farming system. Promotion of the organic farming system for adoption by more banana growers could enhance the sustainability of the Chinese Taipei banana industry.

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