Salt stress on cotton varieties of distinct salinity tolerance can induce expression of different proteins. Zhong 07, a salt-tolerant variety and Zhong s9612, a salt-sensitive variety, were utilized as experimental ma...Salt stress on cotton varieties of distinct salinity tolerance can induce expression of different proteins. Zhong 07, a salt-tolerant variety and Zhong s9612, a salt-sensitive variety, were utilized as experimental materials. The leaves of trefoil seedlings treated with or without 0.4% NaCl for 24 h were harvested for whole-protein extraction. Two-dimensional technology, combined with mass spectroscopy (MS) analysis and protein database searching, was employed to detect differentially expressed proteins and determine their identities and biological functions. Compared with the control, Zhong 07 showed 10 differentially expressed proteins under salt stress, of which 6 were upregulated and 4 were downregulated. Meanwhile, 12 differentially expressed proteins were detected in Zhong s9612 under salt stress, of which 10 were upregulated and 2 were downregulated. In the matrix-assisted laser desorption-ionization/time of flight-time of flight/MS analysis, 14 differentially expressed proteins were successfully identified, including the ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisco) large subunit-binding protein subunit alpha (RuBisco α), luminal binding protein (LBP), heat shock protein 70 (Hsp1, 2, 3), pathogenesis-related protein class 10 (PR-10), quinoneoxidoreductase-like protein (QOR), S-adenosylmethioninesyn-thetase (SAMS), enolase (EN), and RuBisco large subunit-binding protein subunit beta (RuBisco β). Cellular function is ultimately executed by proteins, and cotton varieties with different salt tolerance can be influenced by salt stress to various degrees, which can provide certain theoretical foundation for the identification of salt tolerance of cotton varieties. The findings also provide some proteins, such as the RuBisco large subunit binding proteins α and β subunits, OEE2 protein, HSP70, and S-adenosylmethionine synthetase, which can be used as protein markers of salt-to-lerance before- and post-treatment, making a big difference in salt-tolerance identification in cotton.展开更多
Yeast cadmium factor 1 (YCF1), is a member of the ATP-binding cassette (ABC) transporter family. To explore the functions of YCF1 of Saccharomyces cerevisiae (ScYCF1) in the cotton, ScYCF1 was cloned from Saccharomyce...Yeast cadmium factor 1 (YCF1), is a member of the ATP-binding cassette (ABC) transporter family. To explore the functions of YCF1 of Saccharomyces cerevisiae (ScYCF1) in the cotton, ScYCF1 was cloned from Saccharomyces cerevisiae As2.375, with the full-length of 4548 bp. The bioinformatics analysis revealed that the largest component of ScYCF1 protein is leucine (12%). ScYCF1 is alkaline and positive charged, stable, and hydrophilic protein. The predictive secondary structure is mainly composed of α-helix areas, random coils and β-sheets. We constructed the pBI121-ScYCF1:GFP infusion expression vector and verified it by enzyme ingestion. The transient expression results of cotton pollen showed that the green fluorescence phenomenon of three kinds of upland cotton pollen significantly increased after transforming ScYCF1. The salt sensitive material upland cotton CCRI12 was transformed in vivo simultaneously, and the germination ability of trans-ScYCF1-gene T0 seeds was much better than the acceptor material CCRI12 under the stress of 100 mM NaCl saline solution. According to the gene nucleotide sequences, four pairs of primers were designed for molecular detection of T0 generation, and the sequencing results of PCR products of four specific primers evidence that the transgene is successful. Salt tolerance analysis of leaf discs of identified transgenic cotton showed that the chlorophyll content of leaf discs of transgenic cotton was higher than the content of the control cotton under salt stress. ScYCF1 gene was cloned and introduced into cotton, showing that ScYCF1 plays an important role in improving the salt tolerance of cotton.展开更多
文摘Salt stress on cotton varieties of distinct salinity tolerance can induce expression of different proteins. Zhong 07, a salt-tolerant variety and Zhong s9612, a salt-sensitive variety, were utilized as experimental materials. The leaves of trefoil seedlings treated with or without 0.4% NaCl for 24 h were harvested for whole-protein extraction. Two-dimensional technology, combined with mass spectroscopy (MS) analysis and protein database searching, was employed to detect differentially expressed proteins and determine their identities and biological functions. Compared with the control, Zhong 07 showed 10 differentially expressed proteins under salt stress, of which 6 were upregulated and 4 were downregulated. Meanwhile, 12 differentially expressed proteins were detected in Zhong s9612 under salt stress, of which 10 were upregulated and 2 were downregulated. In the matrix-assisted laser desorption-ionization/time of flight-time of flight/MS analysis, 14 differentially expressed proteins were successfully identified, including the ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisco) large subunit-binding protein subunit alpha (RuBisco α), luminal binding protein (LBP), heat shock protein 70 (Hsp1, 2, 3), pathogenesis-related protein class 10 (PR-10), quinoneoxidoreductase-like protein (QOR), S-adenosylmethioninesyn-thetase (SAMS), enolase (EN), and RuBisco large subunit-binding protein subunit beta (RuBisco β). Cellular function is ultimately executed by proteins, and cotton varieties with different salt tolerance can be influenced by salt stress to various degrees, which can provide certain theoretical foundation for the identification of salt tolerance of cotton varieties. The findings also provide some proteins, such as the RuBisco large subunit binding proteins α and β subunits, OEE2 protein, HSP70, and S-adenosylmethionine synthetase, which can be used as protein markers of salt-to-lerance before- and post-treatment, making a big difference in salt-tolerance identification in cotton.
文摘Yeast cadmium factor 1 (YCF1), is a member of the ATP-binding cassette (ABC) transporter family. To explore the functions of YCF1 of Saccharomyces cerevisiae (ScYCF1) in the cotton, ScYCF1 was cloned from Saccharomyces cerevisiae As2.375, with the full-length of 4548 bp. The bioinformatics analysis revealed that the largest component of ScYCF1 protein is leucine (12%). ScYCF1 is alkaline and positive charged, stable, and hydrophilic protein. The predictive secondary structure is mainly composed of α-helix areas, random coils and β-sheets. We constructed the pBI121-ScYCF1:GFP infusion expression vector and verified it by enzyme ingestion. The transient expression results of cotton pollen showed that the green fluorescence phenomenon of three kinds of upland cotton pollen significantly increased after transforming ScYCF1. The salt sensitive material upland cotton CCRI12 was transformed in vivo simultaneously, and the germination ability of trans-ScYCF1-gene T0 seeds was much better than the acceptor material CCRI12 under the stress of 100 mM NaCl saline solution. According to the gene nucleotide sequences, four pairs of primers were designed for molecular detection of T0 generation, and the sequencing results of PCR products of four specific primers evidence that the transgene is successful. Salt tolerance analysis of leaf discs of identified transgenic cotton showed that the chlorophyll content of leaf discs of transgenic cotton was higher than the content of the control cotton under salt stress. ScYCF1 gene was cloned and introduced into cotton, showing that ScYCF1 plays an important role in improving the salt tolerance of cotton.
