Abstract
The leaf curl disease of Jatropha caused by geminiviruses results in heavy economic losses. In the present study, we report the identification of a new strain of a Jatropha leaf curl Gujarat virus (JLCuGV), which encodes six ORFs with each one having RNA silencing suppressor activity. Therefore, three artificial microRNAs (amiRNAs; C1/C4, C2/C3 and V1/V2) were designed employing overlapping regions, each targeting two ORFs of JLCuGV genomic DNA and transformed in tobacco. The C1/C4 and C2/C3 amiRNA transgenics were resistant while V1/V2 amiRNA transgenics were tolerant against JLCuGV. The relative level of amiRNA inversely related to viral load indicating a correlation with disease resistance. The assessment of photosynthetic parameters suggests that the transgenics perform significantly better in response to JLCuGV infiltration as compared to wild type (WT). The metabolite contents were not altered remarkably in amiRNA transgenics, but sugar metabolism and tricarboxylic acid (TCA) cycle showed noticeable changes in WT on virus infiltration. The overall higher methylation and demethylation observed in amiRNA transgenics correlated with decreased JLCuGV accumulation. This study demonstrates that amiRNA transgenics showed enhanced resistance to JLCuGV while efficiently maintaining normalcy in their photosynthesis and metabolic pathways as well as homeostasis in the methylation patterns.
Highlights
The leaf curl disease of Jatropha caused by geminiviruses results in heavy economic losses
The geminivirus infected leaves were collected from J. curcas CP9 accession (IC 565735), from Council of Scientific and Industrial Research (CSIR)-CSMCRI Jatropha plantation fields, Neswad (Gujarat, India)
This study shows that each of the 6 ORFs of the newly isolated Jatropha leaf curl Gujarat virus (JLCuGV) genomic DNA (AC1, AC2, AC3, AC4, AV1 and AV2), independently show strong RNA silencing suppressors (RSSs) activity
Summary
The leaf curl disease of Jatropha caused by geminiviruses results in heavy economic losses. We report the identification of a new strain of a Jatropha leaf curl Gujarat virus (JLCuGV), which encodes six ORFs with each one having RNA silencing suppressor activity. The geminiviruses infect a wide range of economically important crops like cassava, cotton, grain legumes and tomato, causing significant financial losses across the world. The symptoms of the disease include mosaic and curling of leaves and malformed fruits To date, both monopartite and bipartite geminiviruses have been detected in Jatropha from India, Kenya, Nigeria, Dominic Republican, Jamaica and the United States of America (USA)[13]. The artificial miRNA (amiRNA) provide a highly specific alternative to silence the viral transcripts for targeting the RSS for developing disease r esistance[23]
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