In Silico search and biological validation of MicroR171 family related to abiotic stress response in mulberry (Morus alba)

miR171 belongs to a highly conserved and ubiquitously expressed microRNA gene family across species that play a critical role in controlling plant growth and development through the regulation of the miR171-SCL (scarecrow-like proteins) module. There is limited research available on the evolutionary...

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Main Authors: Zhichao Sun, R.M. Saravana Kumar, Jisheng Li, Guiming Yang, Yan Xie
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2022-03-01
Series:Horticultural Plant Journal
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2468014121001370
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author Zhichao Sun
R.M. Saravana Kumar
Jisheng Li
Guiming Yang
Yan Xie
author_facet Zhichao Sun
R.M. Saravana Kumar
Jisheng Li
Guiming Yang
Yan Xie
author_sort Zhichao Sun
collection DOAJ
description miR171 belongs to a highly conserved and ubiquitously expressed microRNA gene family across species that play a critical role in controlling plant growth and development through the regulation of the miR171-SCL (scarecrow-like proteins) module. There is limited research available on the evolutionary relationship and functional diversification of miR171 members. In this study, we identified eight miR171 genes in the mulberry genome by bioinformatics analysis that were subsequently used to compare the evolutionary levels and explore abiotic stress mechanisms mediated by mno-miR171s (Morus notabilis miR171s). The results of phylogenetic analysis showed that the mature mno-miR171 sequences have strong sequence conservation, but their critical sites also exhibit high variation leading to functional diversification. Through quantitative real-time PCR, the expression profile of each mno-miR171 was analyzed under different stress treatments. All mno-miR171s, apart from mno-miR171h, were found to be significantly up-regulated under salt and drought stress conditions. The target genes of mno-miR171a namely, Morus020885 and Morus011800, were predicted and verified using the plural RNA method. 5′-rapid amplification of complementary DNA ends assays further to reveal that the target genes could be degraded by mno-miR171a post-transcriptionally. Overexpression of mno-miR171a in Arabidopsis improved the percentage of seed germination when the seeds were grown in NaCl- and mannitol-containing media. Transgenic plants were observed to grow better under drought conditions. The expression of various stress genes was significantly higher in transgenic plants than in wild type, except ERF11. Taken together, our study confirmed that mno-miR171a enhanced plant resistance to adverse stress environments via the regulation of the SCL targets.
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spelling doaj.art-acc80e12f001465ea1298781e6b233212024-04-16T14:59:37ZengKeAi Communications Co., Ltd.Horticultural Plant Journal2468-01412022-03-0182184194In Silico search and biological validation of MicroR171 family related to abiotic stress response in mulberry (Morus alba)Zhichao Sun0R.M. Saravana Kumar1Jisheng Li2Guiming Yang3Yan Xie4Hebei Universities R & D Center for Sericulture and Specialty Enabling Technologies, Chengde 067000, China; Sericultural Research Insitute, Chengde Medical University, Chengde 067000, ChinaHebei Universities R & D Center for Sericulture and Specialty Enabling Technologies, Chengde 067000, China; Sericultural Research Insitute, Chengde Medical University, Chengde 067000, ChinaHebei Universities R & D Center for Sericulture and Specialty Enabling Technologies, Chengde 067000, China; Sericultural Research Insitute, Chengde Medical University, Chengde 067000, ChinaHebei Universities R & D Center for Sericulture and Specialty Enabling Technologies, Chengde 067000, China; Sericultural Research Insitute, Chengde Medical University, Chengde 067000, ChinaHebei Universities R & D Center for Sericulture and Specialty Enabling Technologies, Chengde 067000, China; Sericultural Research Insitute, Chengde Medical University, Chengde 067000, China; Corresponding author. Tel.: +863142291114miR171 belongs to a highly conserved and ubiquitously expressed microRNA gene family across species that play a critical role in controlling plant growth and development through the regulation of the miR171-SCL (scarecrow-like proteins) module. There is limited research available on the evolutionary relationship and functional diversification of miR171 members. In this study, we identified eight miR171 genes in the mulberry genome by bioinformatics analysis that were subsequently used to compare the evolutionary levels and explore abiotic stress mechanisms mediated by mno-miR171s (Morus notabilis miR171s). The results of phylogenetic analysis showed that the mature mno-miR171 sequences have strong sequence conservation, but their critical sites also exhibit high variation leading to functional diversification. Through quantitative real-time PCR, the expression profile of each mno-miR171 was analyzed under different stress treatments. All mno-miR171s, apart from mno-miR171h, were found to be significantly up-regulated under salt and drought stress conditions. The target genes of mno-miR171a namely, Morus020885 and Morus011800, were predicted and verified using the plural RNA method. 5′-rapid amplification of complementary DNA ends assays further to reveal that the target genes could be degraded by mno-miR171a post-transcriptionally. Overexpression of mno-miR171a in Arabidopsis improved the percentage of seed germination when the seeds were grown in NaCl- and mannitol-containing media. Transgenic plants were observed to grow better under drought conditions. The expression of various stress genes was significantly higher in transgenic plants than in wild type, except ERF11. Taken together, our study confirmed that mno-miR171a enhanced plant resistance to adverse stress environments via the regulation of the SCL targets.http://www.sciencedirect.com/science/article/pii/S2468014121001370MulberrymiR171qRT-PCRSCLArabidopsisStress
spellingShingle Zhichao Sun
R.M. Saravana Kumar
Jisheng Li
Guiming Yang
Yan Xie
In Silico search and biological validation of MicroR171 family related to abiotic stress response in mulberry (Morus alba)
Horticultural Plant Journal
Mulberry
miR171
qRT-PCR
SCL
Arabidopsis
Stress
title In Silico search and biological validation of MicroR171 family related to abiotic stress response in mulberry (Morus alba)
title_full In Silico search and biological validation of MicroR171 family related to abiotic stress response in mulberry (Morus alba)
title_fullStr In Silico search and biological validation of MicroR171 family related to abiotic stress response in mulberry (Morus alba)
title_full_unstemmed In Silico search and biological validation of MicroR171 family related to abiotic stress response in mulberry (Morus alba)
title_short In Silico search and biological validation of MicroR171 family related to abiotic stress response in mulberry (Morus alba)
title_sort in silico search and biological validation of micror171 family related to abiotic stress response in mulberry morus alba
topic Mulberry
miR171
qRT-PCR
SCL
Arabidopsis
Stress
url http://www.sciencedirect.com/science/article/pii/S2468014121001370
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