A survey of petal-specific proteomes of soybean(Glycine max(L.) Merr[Non-italic].) was conducted comparing protein expression profiles in different petals. Two-dimensional polyacrylamide gel electrophoresis reference ...A survey of petal-specific proteomes of soybean(Glycine max(L.) Merr[Non-italic].) was conducted comparing protein expression profiles in different petals. Two-dimensional polyacrylamide gel electrophoresis reference maps of protein extracts from standard petals(SP), lateral wings(LW), keel petals(KP), and reproductive organs(RO)(a mixture of stamen and carpel) were obtained. Protein expression in the three petal types was compared using Image Master TM 2 D platinum 6.0 software. This indicated that the proportion of homologous proteins between SP and LW was 59.27%, between SP and KP was 61.48%, and between LW and KP was 60.05%. Within a mass range of 6.5-200.0 ku and pH 4.0-7.0, approximately 590, 646, 544, and 700 protein spots were detected in SP, LW, KP, and RO, respectively. A total of 82 differentially expressed proteins were detected. Sixty-four of these detected spots were differentially expressed and showed more than 2-fold changes in abundance; of these 64 proteins, 26 showed increased expression and 38 showed decreased expression. Among these spots, single organ-specific proteins were also identified.They were ID 49(60.9 ku), ID 45(50.0 ku), and ID 46(40.5 ku) in RO, ID 98(42.0 ku) in SP, and ID 05(29.0 ku) in KP. A total of 14 protein spots from 82 differentially expressed proteins were identified with LC-MS/MS. Further protein identification was conducted using the SwissProt and NCBInr databases. The identified proteins and their putative functions were discussed further. This was the first study reporting the comparison of petal protein profiles of soybean florets using proteomics tools.展开更多
Soybean （Glycine max （L.） Merr.） is a major crop that provides plant-origin protein and oil for humans and livestock. Al- though the soybean vegetative tissues and seeds provide a major source of high-quality prot...Soybean （Glycine max （L.） Merr.） is a major crop that provides plant-origin protein and oil for humans and livestock. Al- though the soybean vegetative tissues and seeds provide a major source of high-quality protein, they suffer from low con- centration of an essential sulfur-containing amino acid, methionine, which significantly limits their nutritional quality. The level of methionine is mainly controlled by the first unique enzyme of methionine synthesis, cystathione y-synthase （CGS）. Aiming to elevate methionine level in vegetative tissues and seeds, we constitutively over-expressed a feedback-insensitive Arabidopsis CGS （AtD-CGS） in soybean cultivars, Zigongdongdou （ZD） and Jilinxiaoli 1 （JX）. The levels of soluble methionine increased remarkably in leaves of transgenic soybeans compared to wild-type plants （6.6- and 7.3-fold in two transgenic ZD lines, and 3.7-fold in one transgenic JX line）. Furthermore, the total methionine contents were significantly increased in seeds of the transgenic ZD lines （1.5- to 4.8-fold increase） and the transgenic JX lines （1.3- to 2.3-fold increase） than in the wild type. The protein contents of the transgenic soybean seeds were significantly elevated compared to the wild type, suggesting that the scarcity of methionine in soybeans may limit protein accumulation in soybean seeds. The increased protein content did not alter the profile of major storage proteins in the seeds. Generally, this study provides a promising strategy to increase the levels of methionine and protein in soybean through the breeding programs.展开更多
基金Supported by Harbin Science and Technology Bureau(2016RQYXJ018,2017RAQXJ104)the Key Laboratory of Soybean Biology in the Chinese Ministry of Education,Northeast Agricultural University(SB17A01)+3 种基金the National Natural Science Foundation of China(31801386)Heilongjiang Natural Science Foundation(LC2018008)Heilongjiang General Young Innovative Talents Training Plan(UNPYSCT-2018158)Certificate of China Postdoctoral Science Foundation Grant(2018M641839)
文摘A survey of petal-specific proteomes of soybean(Glycine max(L.) Merr[Non-italic].) was conducted comparing protein expression profiles in different petals. Two-dimensional polyacrylamide gel electrophoresis reference maps of protein extracts from standard petals(SP), lateral wings(LW), keel petals(KP), and reproductive organs(RO)(a mixture of stamen and carpel) were obtained. Protein expression in the three petal types was compared using Image Master TM 2 D platinum 6.0 software. This indicated that the proportion of homologous proteins between SP and LW was 59.27%, between SP and KP was 61.48%, and between LW and KP was 60.05%. Within a mass range of 6.5-200.0 ku and pH 4.0-7.0, approximately 590, 646, 544, and 700 protein spots were detected in SP, LW, KP, and RO, respectively. A total of 82 differentially expressed proteins were detected. Sixty-four of these detected spots were differentially expressed and showed more than 2-fold changes in abundance; of these 64 proteins, 26 showed increased expression and 38 showed decreased expression. Among these spots, single organ-specific proteins were also identified.They were ID 49(60.9 ku), ID 45(50.0 ku), and ID 46(40.5 ku) in RO, ID 98(42.0 ku) in SP, and ID 05(29.0 ku) in KP. A total of 14 protein spots from 82 differentially expressed proteins were identified with LC-MS/MS. Further protein identification was conducted using the SwissProt and NCBInr databases. The identified proteins and their putative functions were discussed further. This was the first study reporting the comparison of petal protein profiles of soybean florets using proteomics tools.
基金supported by the Major Science and Technology Projects of China (2014ZX08004-003,2014ZX08010-004)the Natural Science Foundation of China (31471571)+2 种基金the earmarked fund for China Agriculture Research System (CARS-04)the Agricultural Science and Technology Innovation Project of CAASIsrael Science Foundation (ISF grant 231-09)
文摘Soybean （Glycine max （L.） Merr.） is a major crop that provides plant-origin protein and oil for humans and livestock. Al- though the soybean vegetative tissues and seeds provide a major source of high-quality protein, they suffer from low con- centration of an essential sulfur-containing amino acid, methionine, which significantly limits their nutritional quality. The level of methionine is mainly controlled by the first unique enzyme of methionine synthesis, cystathione y-synthase （CGS）. Aiming to elevate methionine level in vegetative tissues and seeds, we constitutively over-expressed a feedback-insensitive Arabidopsis CGS （AtD-CGS） in soybean cultivars, Zigongdongdou （ZD） and Jilinxiaoli 1 （JX）. The levels of soluble methionine increased remarkably in leaves of transgenic soybeans compared to wild-type plants （6.6- and 7.3-fold in two transgenic ZD lines, and 3.7-fold in one transgenic JX line）. Furthermore, the total methionine contents were significantly increased in seeds of the transgenic ZD lines （1.5- to 4.8-fold increase） and the transgenic JX lines （1.3- to 2.3-fold increase） than in the wild type. The protein contents of the transgenic soybean seeds were significantly elevated compared to the wild type, suggesting that the scarcity of methionine in soybeans may limit protein accumulation in soybean seeds. The increased protein content did not alter the profile of major storage proteins in the seeds. Generally, this study provides a promising strategy to increase the levels of methionine and protein in soybean through the breeding programs.
