High Efficiency Transformation of Banana [Musa acuminata L. cv. Matti (AA)] for Enhanced Tolerance to Salt and Drought Stress Through Overexpression of a Peanut Salinity-Induced Pathogenesis-Related Class 10 Protein

Abstract

Bananas and plantains (Musa spp. L.) are important subsistence crops and premium export commodity in several countries, and susceptible to a wide range of environmental and biotic stress conditions. Here, we report efficient, rapid, and reproducible Agrobacterium-mediated transformation and regeneration of an Indian niche cultivar of banana [M. acuminata cv. Matti (AA)]. Apical meristem-derived highly proliferative multiple shoot clump (MSC) explants were transformed with the Agrobacterium strain EHA105 harboring a binary vector pCAMBIA-1301 carrying hptII and uidA. Sequential agro-infiltration (10 min, 400 mmHg), infection (additional 35 min, Agrobacterium density A 600 = 0.8) and co-cultivation (18 h) regimen in 100 µM acetosyringone containing liquid medium were critical factors yielding high transformation efficiency (~81 %) corroborated by transient GUS expression assay. Stable transgenic events were recovered following two cycles of meristem initiation and selection on hygromycin containing medium. Histochemical GUS assay in several tissues of transgenic plants and molecular analyses confirmed stable integration and expression of transgene. The protocol described here allowed recovery of well-established putative transgenic plantlets in as little as 5 months. The transgenic banana plants could be readily acclimatized under greenhouse conditions, and were phenotypically similar to the wild-type untransformed control plants (WT). Transgenic plants overexpressing Salinity-Induced Pathogenesis-Related class 10 protein gene from Arachis hypogaea (AhSIPR10) in banana cv. Matti (AA) showed better photosynthetic efficiency and less membrane damage (P < 0.05) in the presence of NaCl and mannitol in comparison to WT plants suggesting the role of AhSIPR10 in better tolerance of salt stress and drought conditions.

Preliminary assessment on Agrobacterium-mediated Transformation of Dendrobium Broga Giant orchids PLBs

“…Optimal Agrobacterium tumefaciens density will enhance the T-DNA transfer capacity from the bacterium to the host. Similar to the results reported, bacterial density of 0.8 units at OD 600nm were documented to be are optimal for Aspergillus terreus , banana (Rustagi et al, 2015), and orchid . Contrarily, lower density of Agrobacterium suspension at OD 600nm has been recorded to enhance transformation frequency.…”

Respuesta de cultivares de Musa spp. al estrés hídrico in vitro inducido con polietilenglicol 6000

“…también han informado la presencia de daños a nivel de membranas celulares producto de un estrés hídrico in vitro. Por ejemplo, Moreno-Bermúdez et al (2014), en el cultivar de banano 'Grande naine', Shekhawat & Ganapathi (2014) y Tak et al (2017), en el cultivar de plátano 'Rasthali' (AAB) y Chai et al (2005), y Rustagi et al (2015), en los cultivares de banano 'Berangan' y 'Matti' (AA) respectivamente. En todos estos casos se relacionó el aumento en los contenidos de MDA con una sensibilidad al estrés, y aquellas plantas cuyos valores de MDA no se incrementaron de manera significativa fueron consideradas más tolerantes.…”

“…Algunos de ellos son la altura de las plantas, la longitud de las raíces, el número de brotes por explante, las masas fresca y seca, el contenido relativo de agua, y los contenidos de prolina, malondial-dehido y peróxido de hidrógeno entre otros Rustagi et al, 2015;Tak et al, 2017).…”

Overexpression of RSOsPR10, a root-specific rice PR10 gene, confers tolerance against drought stress in rice and drought and salt stresses in bentgrass

“…Tobacco plants overexpressing the Arachis hypogaea AhPR10 gene exhibited enhanced tolerance against salinity and drought stresses (Jain et al 2012). Similarly, overexpression of the peanut AhSIPR10 gene in banana also resulted in increased tolerance against salinity and drought stresses (Rustagi et al 2015). These reports confirm that the constitutive production of PR10 proteins from various plant sources can increase tolerance against environmental stresses in tissue-and growth stage-specific manners.…”