Expression of a rice chitinase gene in transgenic banana (‘Gros Michel’, AAA genome group) confers resistance to black leaf streak disease

Abstract

Transgenic banana ( Musa acuminata ‘Gros Michel’) integrating either of two rice chitinase genes was generated and its resistance to Black Leaf Streak disease caused by the fungus Mycosphaerella fijiensis was tested using a leaf disk bioassay. PCR screening indicated the presence of the hpt selectable marker gene in more than 90 % of the lines tested, whereas more than three quarters of the lines contained the linked rice chitinase gene resulting in a co-transformation frequency of at least 71.4 %. Further, a unique stable integration of the transgenes in each line revealed some false negative PCR results and the expected co-transformation frequency of 100 %. The transgene insert number per line ranged from 1 to 5 and single transgene insert lines (25 % of all) were identified. Considerable delay in disease development (up to 63 days post-incoculation) over a monitoring period of 108 days occurred in nine lines with extracellularly targeted chitinase out of 17 transgenic lines tested and their necrotic leaf area decreased by 73–94 % compared to the untransformed susceptible control line. Finally, correlation between symptom development and rice chitinase expression was confirmed in two lines by Western analysis. The potential of rice chitinase genes to enhance resistance against M. fijiensis in banana was demonstrated as well as the usefulness of the leaf disk bioassay for early disease screening in transgenic banana lines. Electronic supplementary material The online version of this article (doi:10.1007/s11248-012-9631-1) contains supplementary material, which is available to authorized users.

Susceptibility of Wheat Varieties to Soil-Borne <i>Rhizoctonia</i> Infection

“…The breading is the most promising measure to improve the tolerance to soil-borne pathogen complex, because the use of any xenobiotic has adverse effects on soil biota Hordeum vulgare rip30 increased tolerance (potato) [71] Hordeum vulgare chitinase increased tolerance (tobacco) [72] Pennisetum glaucum lipid transfer protein antifungal [73] Triticum sp. puroindoline increased tolerance (rice) [74] Celastrus hypoleucus pristimerin inhibiting the formation of infective body [75] Celastrus hypoleucus celastrol inhibiting the formation of infective body [75] Prokaryote 5-enolpyruvoyl-shikimate-3-phosphate synthetase Increased tolerance (wheat/Puccinia) [76] Oryza sativa thaumatin like protein increased tolerance (rice) [70] Oryza sativa OsPR-4b gene encoding pathogenesis related protein enhanced resistance [77] Solanum tuberosum Potide G proteinase inhibitor [78] Bacillus subtilis Iturin A Antifungal [79] Bacillus subtilis flagellin Antifungal [80] Raphanus sativus defensin increased tolerance (wheat) [81] Solanum tuberosum Snakin 1 enhanced resistance [82] Dasypyrum villosum unknown tolerance to AG 8 [83] Oryza sativa Rice chitinase increased tolerance (Musa/Mycosphaerella) [84] Oryza sativa Rice chitinase increased tolerance (Eleusine/Magnaporthe) [85] Tichoderma harzianum glucanase inhibiting the formation of infective body [86] Tephrosia villosa defensin increased tolerance (tobacco) [87] Arabidopsis thaliana NADPH oxydase induced resistance [88] Oryza sativa ACCA synthase induced resistance [89] as well as can predispose host plant to pathogen [62,63].…”

Transgenic Chili PossessingBaculovirus Chitinase GeneExhibitsin vitroFungal Inhibition

“…Variations in the chitinase activities in plants transformed with chitinase gene have been recorded (Tohidfar et al 2005;Yamamoto et al 2000;Deroles and Gardner 1988). Correlation between disease symptom development and expression of chitinase gene has been reported (Hu et al 2013;Kovacs et al 2013;Prasad et al 2013). Thus, a greater percentage of plants, about 14% (4/28) of line C4 (expressing 4.8x higher endochitinase activity than C7) exhibited higher degree of tolerance as compared with only 4% (2/55) lines of C7 exhibiting some degree of tolerance (Tables 5-6, Figures 10-12).…”

“…Chitinase is an excellent candidate for this purpose because it can attack pathogen structure directly (Arlorio et al 1992;Ji and Kuc 1996). The antifungal activity of chitinases has long been known and has been successfully used for developing transgenic crops with enhanced resistance to several fungi (Broglie et al 1991;Lorito et al 1998;Punja 2004;Saiprasad et al 2009;Kovacs et al 2013). Chitinase genes have originated from several sources, including bacteria, baculovirus, plants, and fungi.…”

Next Generation Plant Biotechnology

“…• Bananas that are resistant to fungal wilt (Panama wilt) and black leaf streak diseases, and exhibit increased β-carotene and iron (Aravanityonnis et al 2008;Kovács et al 2013;Cressey 2013); • Corn with enhanced levels of multivitamins, including β-carotene (provitamin A), ascorbate (vitamin C), and folate (vitamin B9) (Naqvi et al 2009);…”