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Blast resistance gene Pi54 over-expressed in rice to understand its cellular and sub-cellular localization and response to different pathogens

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Title Blast resistance gene Pi54 over-expressed in rice to understand its cellular and sub-cellular localization and response to different pathogens
 
Creator Singh, Jyoti
Gupta, Santosh Kumar
Devanna, B. N.
Singh, Sunil
Upadhyay, Avinash
Sharma, Tilak R.
 
Subject rice
Biotic
Molecular engineering in plants
Blast resistance gene
 
Description Accepted date: 31 December 2019
Rice blast resistance gene, Pi54 provides broad-spectrum resistance against diferent strains of
Magnaporthe oryzae. Understanding the cellular localization of Pi54 protein is an essential step towards
deciphering its place of interaction with the cognate Avr-gene. In this study, we investigated the subcellular localization of Pi54 with Green Fluorescent Protein (GFP) as a molecular tag through transient
and stable expression in onion epidermal cells (Allium cepa) and susceptible japonica cultivar rice Taipei
309 (TP309), respectively. Confocal microscopy based observations of the onion epidermal cells revealed
nucleus and cytoplasm specifc GFP signals. In the stable transformed rice plants, GFP signal was recorded
in the stomata, upper epidermal cells, mesophyll cells, vascular bundle, and walls of bundle sheath and
bulliform cells of leaf tissues. These observations were further confrmed by Immunocytochemical studies.
Using GFP specifc antibodies, it was found that there was sufcient aggregation of GFP::Pi54protein in
the cytoplasm of the leaf mesophyll cells and periphery of the epidermal cells. Interestingly, the transgenic
lines developed in this study could show a moderate level of resistance to Xanthomonas oryzae and
Rhizoctonia solani, the causal agents of the rice bacterial blight and sheath blight diseases, respectively.
This study is a frst detailed report, which emphasizes the cellular and subcellular distribution of the broad
spectrum blast resistance gene Pi54 in rice and the impact of its constitutive expression towards resistance
against other fungal and bacterial pathogens of rice.
TR Sharma is grateful to Department of Biotechnology Govt. of India for financial help and DST for JC Bose National Fellowship. We are thankful to Dr. Rithu Rai for sharing X. oryzae strains used in this study. Authors are thankful to Dr. Praveen Verma, Scientist NIPGR, New Delhi for their valuable suggestions in FM 4–64 staining. We also thank the in-charge of National Phytotron Facility IARI, New Delhi for providing facilities to maintain transgenic rice lines and conducting phenotyping experiments.
 
Date 2020-04-06T15:39:43Z
2020-04-06T15:39:43Z
2020
 
Type Article
 
Identifier Scientific Reports, 10: 5243
2045-2322
https://doi.org/10.1038/s41598-020-59027-x
https://www.nature.com/articles/s41598-020-59027-x
http://223.31.159.10:8080/jspui/handle/123456789/1051
 
Language en_US
 
Format application/pdf
 
Publisher Springer Nature Limited