|
|
|
(3.129.7.43)
|
|
Users online: 18366
|
|
|
|
|
|
|
Ijournet
|
|
|
|
|
|
|
An economical approach for production of double stranded RNA for RNAi gene silencing in management of Meloidogyne incognita Chatterjee Madhurima, Rao Uma* Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi-110 012 *umanema@gmail.com; umarao@iari.res.in
Online published on 7 April, 2020. Abstract Meloidogyne incognita, a sedentary and obligate endoparasite is one of the most economically important nematodes globally. Non-chemical based nematode control strategies have gained significant importance due to either nonavailability or restrictions on use of synthetic chemical nematicides in many countries. In this direction, genomics based molecular approaches like RNA interference (RNAi) gene silencing either by in vitro or in planta delivery of double stranded RNA (dsRNA) has been established as a very promising strategy for its management of M. incognita. In vitro RNAi required the synthesis of dsRNA by transcription using expensive commercially available kits. In the present study, it employed a bacterial based approach to synthesize dsRNA of Miflp-14 gene, as an alternative lowcost method using Escherichia coli strain HT115. This alternate approach is convenient to synthesize large quantities of dsRNA and more economical compared to that of synthesis using commercial kits. For this, it was cloned a 284 bp of Miflp-14 fragment into RNAi vector L4440, bidirectionally flanked by T7 promoter and transformed into RNaseIII deficient E. coli strain HT115 and synthesized dsRNA of Miflp-14 using the commercial kit. M. incognita J2s were soaked in the dsRNA synthesized by both methods for inducing RNAi of Miflp-14. RNAi silencing was confirmed by qRT-PCR analysis. The extent of down regulation of Miflp-14 expression obtained through soaking of M. incognita J2s in bacterially expressed dsRNA was comparable to that of in vitro transcription kit-synthesized dsRNA. Top Keywords Double stranded RNA (dsRNA), FMRF-amide like peptide gene (flp-14), Meloidogyne incognita, RNA interference. Top | |
|
|
|
|
║ Site map
║
Privacy Policy ║ Copyright ║ Terms & Conditions ║
|
|
|
751,718,738 visitor(s) since 30th May, 2005.
|
|
All rights reserved. Site designed and maintained by DIVA ENTERPRISES PVT. LTD..
|
|
Note: Please use Internet Explorer (6.0 or above). Some functionalities may not work in other browsers.
|