Over-expression of PnSMO1.1 Gene Improves Salinity-stress Resistance of Transgenic Pharbitis nil Seedlings

doi:10.7668/hbnxb.20192731

Abstract

Abstract:

Application of some hormonal signaling compounds,as brassinolides and their derivatives,could significantly improve the salt stress resistance in plants.The purpose of present work was to test whether the over-expressing of Methylsterol monooxygenase gene(SMO),a key gene coding a bio-synthesizing enzyme of sterol in plants,could promote the salt-stress tolerance of target plants.PnSMO1.1,gene encoding of methylsterol monooxygenase in the plant species of Pharbitis nil was firstly cloned and then used as target gene for following genetic transformation process,while wild-type Pharbitis nil seedlings were used as the receptor plants for constructing of the transgenic lines which over-expressed PnSMO1.1 genes.In this work,the PnSMO1.1 gene transformed Pharbitis nil lines were constructed via an ovary injection transformation method.Plantlets from individual PCR identified transgenic plant lines were used as materials to detect vegetative growth figures and some pivotal physiological indicators,for instance,contents of malondialdehyde (MDA),relative conductivity,as well as castasterone and 6-deoxo-castasterone contents in cells under stresses with gradient NaCl conditions varied from 0—200 mmol/L.The results showed that the over-expression PnSMO1.1 significantly improved the relative growth of roots and hypocotyls in transformants than in wild-type (WT) or vacant plasmid transformed control (BL) plants under 100—250 mmol/L NaCl stresses.Compared to WT and BL seedlings,significantly higher accumulation of 6-deoxo-castasterone,but lower relative conductivity (rEC) values,castasterone accumulation or MDA contents were found in transgenic lines under various NaCl stresses.Stresses such as salinity,drought and freezing temperatures,had severely suppressed the vegetative growth of plants,as well as their yields.These results highlighted that over-expression of the PnSMO1.1 gene could significantly improve the salinity-stress resistance of transgenic plants by delicately adjusting the dynamic homeostasis of brassinolides in cells,and protecting the structural integrity of plasma membrane.

Key words: Pharbitis nil, PnSMO1.1, Genetic transformation, NaCl, Salt stress, Ovary injection

TANG Long, ZHAO Yuwei. Over-expression of PnSMO1.1 Gene Improves Salinity-stress Resistance of Transgenic Pharbitis nil Seedlings[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(2): 1-8. doi: 10.7668/hbnxb.20192731.

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Figures/Tables 7

Fig.1 Diagram of T-DNA region in pRI-PnSMO1.1 plasmid

Fig.2 PCR identification of T₀ kanamycin resistant plants
M.DL5000 DNA Marker;PC.PCR positive control with pRI-PnSMO1.1.plasmid DNA as templates;NC.PCR blank control with genomic DNA of pRI 101-AN vacant vector transformed plant lines as template;DC.PCR results with PnSMO1.1 cDNA as templates;S₁—S₈.PCR amplification results with genomic DNA from different PnSMO1.1 gene overexpression lines;2—12.PCR amplification results of kanamycin resistance gene NPT Ⅱ;14—24.PCR amplification results of target gene PnSMO1.1.

Fig.3 Relative growth of root length of different plants under treatment of various NaCl concentrations
Different capital letters indicate significant differences among groups based on the concentration of NaCl;Different lowercase letters indicate significant differences among groups of plants.The same as Fig.4—7.

Fig.4 Relative growth of hypocotyl length of different plants under treatment of various NaCl concentrations

Fig.5 The content of MDA in leaves of different plants under treatment of various NaCl concentrations

Fig.6 The relative conductivity in leaves of different plants under treatment of various NaCl concentrations

Fig.7 The content of brassinolide in leaves of different plants under treatment of various NaCl concentrations

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