Development of SCAR marker specific to non-toxic Jatropha curcas L. and designing a novel multiplexing PCR along with nrDNA ITS primers to circumvent the false negative detection

Handle URI:
http://hdl.handle.net/10754/561774
Title:
Development of SCAR marker specific to non-toxic Jatropha curcas L. and designing a novel multiplexing PCR along with nrDNA ITS primers to circumvent the false negative detection
Authors:
Mastan, Shaik G.; Pamidimarri, D. V N N Sudheer; Rahman, Hifzur; Reddy, Muppala P.; Chikara, Jitendra
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. © 2011 Springer Science+Business Media, LLC.
KAUST Department:
Center for Desert Agriculture
Publisher:
Springer Nature
Journal:
Molecular Biotechnology
Issue Date:
10-May-2011
DOI:
10.1007/s12033-011-9415-5
PubMed ID:
21556845
Type:
Article
ISSN:
10736085
Sponsors:
We are thankful to Council for Scientific and Industrial Research, New Delhi, India, for its financial support and for research associate fellowship. We would like to thank Professor B. N. Tiwari, Head, School of biotechnology, for his kind support and encouragement.
Appears in Collections:
Articles; Center for Desert Agriculture

Full metadata record

DC FieldValue Language
dc.contributor.authorMastan, Shaik G.en
dc.contributor.authorPamidimarri, D. V N N Sudheeren
dc.contributor.authorRahman, Hifzuren
dc.contributor.authorReddy, Muppala P.en
dc.contributor.authorChikara, Jitendraen
dc.date.accessioned2015-08-03T09:04:18Zen
dc.date.available2015-08-03T09:04:18Zen
dc.date.issued2011-05-10en
dc.identifier.issn10736085en
dc.identifier.pmid21556845en
dc.identifier.doi10.1007/s12033-011-9415-5en
dc.identifier.urihttp://hdl.handle.net/10754/561774en
dc.description.abstractJatropha 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. © 2011 Springer Science+Business Media, LLC.en
dc.description.sponsorshipWe are thankful to Council for Scientific and Industrial Research, New Delhi, India, for its financial support and for research associate fellowship. We would like to thank Professor B. N. Tiwari, Head, School of biotechnology, for his kind support and encouragement.en
dc.publisherSpringer Natureen
dc.subjectBiofuelen
dc.subjectJatropha curcasen
dc.subjectMultiplex PCRen
dc.subjectNon-toxic genotypeen
dc.subjectSCARen
dc.titleDevelopment of SCAR marker specific to non-toxic Jatropha curcas L. and designing a novel multiplexing PCR along with nrDNA ITS primers to circumvent the false negative detectionen
dc.typeArticleen
dc.contributor.departmentCenter for Desert Agricultureen
dc.identifier.journalMolecular Biotechnologyen
dc.contributor.institutionDiscipline of Wasteland Research, Central Salt and Marine, Chemicals Research Institute (CSIR)3, Bhavnagar, Gujarat 364002, Indiaen
dc.contributor.institutionGuru Ghasidas Vishwavidyala (A Central University), Koni, Bilaspur, Chhattisgarh 495009, Indiaen
kaust.authorReddy, Muppala P.en

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