NDLI: Enhancement of Trichoderma endochitinase secretion in tobacco cell cultures using an α-amylase signal peptide

Content Provider	Springer Nature Link
Author	Kumari, Archana Sharma, Gaurav Bhat, Sumangala Bhat, Ramesh S. Krishnaraj, P. U. Kuruvinashetti, M. S.
Copyright Year	2011
Abstract	An α-amylase signal peptide from rice was synthesized and fused with endochitinase (ech42) gene cloned from Trichoderma virens. The chimeric gene was designated as PSPα-amyech42, and this was transferred to a plant transformation vector, referred to as pMASGK. Leaf explants of tobacco cv White Burley were co-cultivated with Agrobacterium tumefaciens strain LBA4404 carrying ech42 with its own signal peptide(ech42SP) and PSPα-amyech42(pMASGK) separately. Putative transformants were selected on Murashige and Skoog (MS) medium, supplemented with 1 mg/l bezyladenine (BA), 0.5 mg/l naphthalene acetic acid(NAA), and containing 200 mg/l kanamycin and 200 mg/l cefotaxime. Transformation was further confirmed by PCR with specific primers and Southern blot hybridization. Endochitinase secretion was quantified in 1-week-old cell suspension cultures obtained from 3-week-old callus cultures of transformants carrying PSPα-amyech42, transformants with ech42 and of control (untransformed) plant. Callus cultures of PSPα-amyech42 showed higher endochitinase activity (9–12 times) than those carrying ech42SP (7–8 times) in both medium and cell extracts. Media collected (200 μg of total protein) from PSPα-amyech42 suspension cultures in Potato Dextrose Agar plates showed growth inhibition of 73 and 53% against Sclerotium rolfsii and Rhizoctonia bataticola, respectively, whereas media collected (200 μg of total protein) from ech42SP suspension culture showed inhibition of 14 and 24% against Sclerotium rolfsii and Rhizoctonia bataticola, respectively.
Starting Page	215
Ending Page	224
Page Count	10
File Format	PDF
ISSN	01676857
Journal	Plant Cell, Tissue and Organ Culture (PCTOC)
Volume Number	107
Issue Number	2
e-ISSN	15735044
Language	English
Publisher	Springer Netherlands
Publisher Date	2011-06-09
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Endochitinase α-Amylase Trichoderma Tobacco Secretion Plant Physiology Plant Genetics & Genomics Plant Pathology Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Horticulture

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ENHANCEMENT OF Trichoderma ENDOCHITINASE SECRETION IN TOBACCO CELL CULTURE USING α-amylase SIGNAL PEPTIDE

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Enhancement of Trichoderma endochitinase secretion in tobacco cell cultures using an α-amylase signal peptide

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