Apple scab resistance genes, HcrVf1 and HcrVf2, were isolated including their native promoter, coding and terminator sequences. Two fragment lengths (short and long) of the native gene promoters and the strong apple rubisco gene promoter (PMdRbc) were used for both HcrVf genes to test their effect on expression and phenotype. The scab susceptible cultivar ‘Gala’ was used for plant transformations and after selection of transformants, they were micrografted onto apple seedling rootstocks for scab disease tests. Apple transformants were also tested for HcrVf expression by quantitative RT-PCR (qRT-PCR). For HcrVf1 the long native promoter gave significantly higher expression that the short one; in case of HcrVf2 the difference between the two was not significant. The apple rubisco gene promoter proved to give the highest expression of both HcrVf1 and HcrVf2. The top four expanding leaves were used initially for inoculation with monoconidial isolate EU-B05 which belongs to race 1 of V. inaequalis. Later six other V. inaequalis isolates were used to study the resistance spectra of the individual HcrVf genes. The scab disease assays showed that HcrVf1 did not give resistance against any of the isolates tested regardless of the expression level. The HcrVf2 gene appeared to be the only functional gene for resistance against Vf avirulent isolates of V. inaequalis. HcrVf2 did not provide any resistance to Vf virulent strains, even not in case of overexpression. In conclusion, transformants carrying the apple-derived HcrVf2 gene in a cisgenic as well as in an intragenic configuration were able to reach scab resistance levels comparable to the Vf resistant control cultivar obtained by classical breeding, cv. ‘Santana’.Electronic supplementary materialThe online version of this article (doi:10.1007/s11103-011-9749-1) contains supplementary material, which is available to authorized users.
The development of crop varieties with higher nitrogen use efficiency is crucial for sustainable crop production. Combining high-throughput genotyping and phenotyping will expedite the discovery of novel alleles for breeding crop varieties with higher nitrogen use efficiency. Digital and hyperspectral imaging techniques can efficiently evaluate the growth, biophysical, and biochemical performance of plant populations by quantifying canopy reflectance response. Here, these techniques were used to derive automated phenotyping of indicator biomarkers, biomass and chlorophyll levels, corresponding to different nitrogen levels. A detailed description of digital and hyperspectral imaging and the associated challenges and required considerations are provided, with application to delineate the nitrogen response in wheat. Computational approaches for spectrum calibration and rectification, plant area detection, and derivation of vegetation index analysis are presented. We developed a novel vegetation index with higher precision to estimate chlorophyll levels, underpinned by an image-processing algorithm that effectively removed background spectra. Digital shoot biomass and growth parameters were derived, enabling the efficient phenotyping of wheat plants at the vegetative stage, obviating the need for phenotyping until maturity. Overall, our results suggest value in the integration of high-throughput digital and spectral phenomics for rapid screening of large wheat populations for nitrogen response.
Delaying leaf senescence in plants, especially under water stress conditions, can help to maintain the remobilization of stored nutrients in source–sink relationships, thus leading to improved crop yields. Leaf senescence can be delayed by plant hormones such as cytokinin. Here, the Isopentenyl transferase (IPT) gene, encoding a cytokinin biosynthesis enzyme, driven by a modified AtMYB32xs promoter was transformed into wheat. Transgenic wheat plants exhibited delayed leaf senescence, retaining chlorophyll for longer under controlled environment conditions. Selected independent transgenic events and their corresponding nulls were grown under field conditions for two consecutive years under well-watered and water stress treatments using automated rainout shelters. Three independent transgenic events had improved canopy green cover, lower canopy temperatures, and higher leaf water potential than their respective non-transgenic nulls, with no abnormality in morphology and phenology. Increased grain yield was observed in transgenic events under both water treatments, with the yield increase more pronounced under water stress (26–42%). These results have shown that delayed leaf senescence using the chimeric transgene AtMYB32xs-p::IPT can be a useful strategy to achieve grain yield gains in wheat and potentially other crops for sustainable food production.
Black root rot, caused by Thielaviopsis basicola, is an important disease in several crops including cotton. We studied the response of Arabidopsis NPR1 (AtNPR1)-expressing cotton lines, previously shown to be highly resistant to a diverse set of pathogens, to a challenge from T. basicola. In four different experiments, we found significant degree of tolerance in the transgenic lines to black root rot. Although transformants showed the typical root discoloration symptoms similar to the wild-type control plants following infection, their roots tended to recover faster and resumed normal growth. Better performance of transgenic plants is reflected by the fact that they have significantly higher shoot and root mass, longer shoot length, and greater number of boll-set. Transcriptional analysis of the defense response showed that the roots of AtNPR1-overexpressing transgenic plants exhibited stronger and faster induction of most of these defense-related genes, particularly PR1, thaumatin, glucanase, LOX1, and chitinase. The results obtained in this investigation provide further support for a broad-spectrum nature of the resistance conferred by overexpression of AtNPR1 gene in cotton.
Apple scab, caused by the fungal pathogen Venturia inaequalis, is one of the most devastating diseases for the apple growing in temperate zones with humid springs and summers. Breeding programs around the world have been able to identify several sources of resistance, the Vf from Malus floribunda 821 being the most frequently used.
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