Development of SCAR Marker Specific to Non-Toxic <Emphasis Type="Italic">Jatropha curcas</Emphasis> L. and Designing a Novel Multiplexing PCR Along with nrDNA ITS Primers to Circumvent the False Negative Detection |
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Authors: | Shaik G Mastan Pamidimarri D V N Sudheer Hifzur Rahman Muppala P Reddy Jitendra Chikara |
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Institution: | (1) Discipline of Wasteland Research, Central Salt and Marine, Chemicals Research Institute (CSIR), Bhavnagar, Gujarat, 364002, India;(2) Guru Ghasidas Vishwavidyala (A Central University), Koni, Bilaspur, 495009, Chhattisgarh, India;(3) Plant Stress Genomics and Technology Center, King Abdullah University of Science and Technology, Thuwal, 23955-6900, Kingdom of Saudi Arabia; |
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Abstract: | Jatropha curcas L., a multipurpose shrub, has acquired significant economic importance for its seed oil which can be converted to biodiesel
an emerging alternative to petro-diesel. In addition to the commercial value, it is also having medicinal and even high nutritional
value to use as animal fodder which is limited due to the toxicity. Development of molecular marker will enable to differentiate
non-toxic from toxic variety of J. curcas in a mixed population and also for quality control since the toxic components of J. curcas has deleterious effect on animals. In the present study, the efforts were made to generate the specific SCAR marker for toxic
and/or non-toxic J. curcas from RAPD markers. Among the markers specific for toxic and non-toxic varieties, four were selected, purified, cloned, sequenced,
and designed primers out of which one set of primers NT-JC/SCAR I/OPQ15-F and R could able to discriminate the non-toxic with
toxic Jatropha by giving expected 430 bp size amplification in non-toxic variety. Furthermore, novel multiplex PCR was designed using the
nrDNA ITS primers to overcome the false negatives. Present work also demonstrates utility of the conserved regions of nrDNA
coding genes in ruling out the artifacts in PCR-like false negatives frequently occur in SCAR due to various reasons. The
specific SCAR markers generated in the present investigation will help to distinguish non-toxic from toxic varieties of J. curcas or vice versa, and isolated marker along with designed multiplex protocol has applications in quality control for selective
cultivation of non-toxic variety and will also assist in breeding and molecular mapping studies. |
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