scholarly journals Cloning, Expression and Genetic Transformation of Sucrose Phosphate Synthase (Sps) Gene in Saccharum Spontaneum L.

2017 ◽  
Vol 59 (2) ◽  
pp. 7-15
Author(s):  
Yun-Wei Zhang ◽  
Yun-Zhuan Zhou ◽  
Hai-Bo Lu ◽  
Deng-Yu Zheng ◽  
Yan-Hua Huang
Keyword(s):  
Sucrose Phosphate Synthase ◽  
Saccharum Spontaneum ◽  
Reading Frame ◽  
Acid Protein ◽  
Significant Difference ◽  
Key Enzyme ◽  
Young Leaves ◽  
Sucrose Phosphate ◽  
Phosphate Synthase

AbstractSucrose phosphate synthase (SPS) is a key enzyme catalyzing sucrose metabolism in plants. In this study, we isolated the SPS cDNA from Saccharum spontaneum and designated as SsSPS (GenBank accession no. MF398541). The full-length of SsSPS cDNA was 4153-bp with an opening reading frame (ORF) of 3132 nucleotides, which encoded a 1043-amino acid protein. The nucleotide sequences alignment showed that it had 98%, 97% and 87% homology with S. officinarum, Setaria italica and Lolium perenne, respectively. Moreover, the SsSPS was detected to express in leaf and stem tissues of S. spontaneum and exhibited a predominant expression in the stem tissue. However, there was no significant difference in the expression level of SsSPS between young leaves and mature ones. Additionally, we generated transgenic S. spontaneum using Agrobacterium-mediated transformation. Our data will provide a valuable foundation for further study of the potential role of SPS in plants.

scholarly journals Effects of CEPA and 1-MCP on flower bud differentiation of apple cv. ‘Nagafu No.2’ grafted on different rootstocks

2018 ◽  
Author(s):  
Wenfang Li ◽  
Baihong Chen ◽  
Juan Mao ◽  
Xinwen Li ◽  
Jing Su ◽  
...  
Keyword(s):  
Sucrose Synthase ◽  
Soluble Sugar ◽  
Sucrose Phosphate Synthase ◽  
Flower Formation ◽  
Flower Bud ◽  
Juvenile Period ◽  
Significant Difference ◽  
Flowering Rate ◽  
Sucrose Phosphate ◽  
Phosphate Synthase

AbstractThe apple (Malus domestica Borkh.) has a relatively long juvenile period which prevent the fruit breeding. The understanding of the flowering system is important to improve breeding efficiency in the apple. In this context, 2-year-old “Fuji” apple cv. “Nagafu No.2” trees that were grafted on dwarf self-rooted rootstock M.26, vigorous rootstock M. sieversii and interstock M.26/M. sieversii, respectively. Spraying with clean water (as controls), 800 mg·L−1 2-Chloroethylphosphonic acid (CEPA) and 2 μL·L−1 1-methylcyclopropene (1-MCP). The results showed that CEPA significantly repressed the vegetative growth attributed to the increase of the ABA and ZT synthesis, and the decrease of IAA synthesis in leaves and buds. However, there was no significant difference or significant inverse effect between 1-MCP and control. Furthermore, CEPA promoted flower formation, increased the flowering rate and advanced the blossom period for 2 days compared with the control, which accompanied by the accumulation of soluble sugar, glucose and sucrose, and the increase of α-amylase (α-AMY) and sucrose phosphate synthase (SPS) activities, and the decrease of the starch contents and sucrose synthase (SS) activities in leaves and buds. However, the blossom period was delayed for 2 days after spraying with 1-MCP. Finally, the expression of TFL1 was significantly repressed while the AP1 was significantly promoted in buds from M.26 and M.26/M. sieversii after spraying with CEPA, while the effect was not significant from M. sieversii. However, the expression levels of TFL1 and AP1 were not significantly different from the control after the application of 1-MCP. In spite of this, CEPA was more susceptible to easy-flowering M26, followed by M26/M. sieversii, and still less susceptible to difficult-flowering rootstock M. sieversii.Abbreviations1-MCP1-methylcyclopropeneα-amylase(α-AMY)ABAabscisic acidCEPA2-Chloroethylphosphonic acidCTKcytokininsETHethyleneGAgibberellinSPSsucrose phosphate synthaseSSsucrose synthaseZTzeatin.

scholarly journals TRANSFORMASI GENETIK DAN EKSPRESI MUTAN SUCROSE PHOSPHATE SYNTHASE PADA TANAMAN TOMAT

2019 ◽  
Vol 6 (1) ◽  
pp. 130
Author(s):  
Suwinda Fibriani ◽  
Inyana Dwi Agustien ◽  
Widhi Dyah Sawitri ◽  
Bambang Sugiharto
Keyword(s):  
Genetic Transformation ◽  
Tomato Plant ◽  
Sucrose Phosphate Synthase ◽  
Transgenic Tomato ◽  
Pcr Analysis ◽  
N Terminus ◽  
Key Enzyme ◽  
Sucrose Biosynthesis ◽  
Sucrose Phosphate ◽  
Phosphate Synthase

Genetic Transformation and Expression of Sucrose Phosphate Synthase Mutant in Tomato Plant ABSTRACTSucrose phosphate synthase (SPS) is a key enzyme responsible for sucrose biosynthesis. In its regulation, SPS activity is modulated by an allosteric effector glucose-6-phosphate (G6P) suggested to have an ability to bind SPS N-terminus domain. To understand the role of N-terminus in regulating SPS, the SPS gene was mutated with the deletion of N-terminus domain (∆N-SPS). The ∆N-SPS gen was transformed into tomato plants with 5% transformation efficiency. Three transgenic tomato plant 4.20, 5.5.1, and 5.10 were obtained and confirmed by PCR analysis. Transgenic tomato expression was characterized by enzymatic analysis. Result showed that the G6P allosteric regulation in transgenic ∆N-SPS had lost and the SPS activity increased by 2-fold compared to non-transgenic plant. This showed that N-terminus domain-deleted SPS could be actively expressed in plant. Keywords: enzyme, genetic transformation, N-terminus domain deletion, sucrose phosphate synthase, tomato ABSTRAKSucrose phosphate synthase (SPS) merupakan enzim kunci yang bertanggung jawab dalam sintesis sukrosa. Dalam regulasinya, aktifitas SPS dipengaruhi oleh alosterik efektor glukosa-6-fosfat (G6P) yang diduga dapat berikatan pada domain N-terminus SPS. Untuk mengetahui peran N-terminus pada regulasi SPS, dilakukan mutasi SPS dengan penghilangan domain N-terminus (∆N-SPS). Gen ∆N-SPS diinsersi pada tanaman tomat melalui transformasi genetik dengan efisiensi transformasi 5%. Tiga tanaman transgenik tomat (event4.20; 5.5.1; dan 5.10) didapatkan dan positif terkonfirmasi melalui analisis PCR. Ekspresi mutan dikarakterisasi melalui analisis enzimatik. Hasil menunjukkan bahwa tanaman tomat transgenik ∆N-SPS tidak dipengaruhi regulasi alosterik G6P dan aktifitas SPS 2 kali lipat lebih tinggi daripada tanaman bukan transgenik. Ini menunjukkan bahwa SPS dengan delesi domain N-terminus dapat terekspresi aktif pada tanaman.  Kata Kunci: delesi domain N-terminus, enzim, sucrose phosphate synthase, tomat, transformasi genetik 

scholarly journals Cloning and Characterization of a Sucrose Phosphate Synthase-encoding Gene from Muskmelon

2007 ◽  
Vol 132 (4) ◽  
pp. 557-562 ◽  
Author(s):  
Xiyan Yu ◽  
Xiufeng Wang ◽  
Jide Fan ◽  
Hongmei Tian ◽  
Chengchao Zheng
Keyword(s):  
Critical Role ◽  
Sucrose Phosphate Synthase ◽  
Transcript Level ◽  
Sucrose Accumulation ◽  
Mature Fruit ◽  
Mrna Accumulation ◽  
Reading Frame ◽  
Rapid Amplification ◽  
Sucrose Phosphate ◽  
Phosphate Synthase

Sucrose phosphate synthase [SPS (EC 2.4.1.14)] is thought to play a critical role in sucrose accumulation in muskmelon (Cucumis melo L.) fruit. A full-length cDNA clone encoding sucrose phosphate synthase was isolated from muskmelon by reverse transcriptase–polymerase chain reaction and rapid amplification of cDNA ends. The clone, designated CmSPS1, contains 3377 nucleotides with an open reading frame of 3162 nucleotides. The deduced 1054 amino acids sequence showed high identities with other plant sucrose phosphate synthases. Northern blot analysis indicated that CmSPS1 was expressed in leaves, stems, and mature fruit, but was not detected in roots or flowers. Moreover, the mRNA accumulation of CmSPS1 started at 25 days after pollination (DAP) and reached highest level in mature fruit. Interestingly, both sucrose content and SPS activity increased dramatically between 20 and 30 DAP during fruit development, suggesting that sucrose accumulation may be linked to the CmSPS1 transcript level in muskmelon fruit.

scholarly journals Sucrose Phosphate Synthase, a Key Enzyme for Sucrose Biosynthesis in Plants

1991 ◽  
Vol 96 (2) ◽  
pp. 473-478 ◽  
Author(s):  
Jean-Michel Bruneau ◽  
Ann C. Worrell ◽  
Bernard Cambou ◽  
Danielle Lando ◽  
Toni A. Voelker
Keyword(s):  
Sucrose Phosphate Synthase ◽  
Key Enzyme ◽  
Sucrose Biosynthesis ◽  
Sucrose Phosphate ◽  
Phosphate Synthase

scholarly journals Comparative analysis of sucrose phosphate synthase (SPS) gene family between Saccharum officinarum and Saccharum spontaneum

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Panpan Ma ◽  
Xingtan Zhang ◽  
Lanping Chen ◽  
Qian Zhao ◽  
Qing Zhang ◽  
...  
Keyword(s):  
Sugar Content ◽  
Expression Patterns ◽  
High Stress ◽  
Sucrose Phosphate Synthase ◽  
Saccharum Officinarum ◽  
Sugar Accumulation ◽  
Saccharum Spontaneum ◽  
Sugarcane Cultivar ◽  
Sucrose Phosphate ◽  
Phosphate Synthase

Abstract Background Sucrose phosphate synthase (SPS) genes play vital roles in sucrose production across various plant species. Modern sugarcane cultivar is derived from the hybridization between the high sugar content species Saccharum officinarum and the high stress tolerance species Saccharum spontaneum, generating one of the most complex genomes among all crops. The genomics of sugarcane SPS remains under-studied despite its profound impact on sugar yield. Results In the present study, 8 and 6 gene sequences for SPS were identified from the BAC libraries of S. officinarum and S. spontaneum, respectively. Phylogenetic analysis showed that SPSD was newly evolved in the lineage of Poaceae species with recently duplicated genes emerging from the SPSA clade. Molecular evolution analysis based on Ka/Ks ratios suggested that polyploidy reduced the selection pressure of SPS genes in Saccharum species. To explore the potential gene functions, the SPS expression patterns were analyzed based on RNA-seq and proteome dataset, and the sugar content was detected using metabolomics analysis. All the SPS members presented the trend of increasing expression in the sink-source transition along the developmental gradient of leaves, suggesting that the SPSs are involved in the photosynthesis in both Saccharum species as their function in dicots. Moreover, SPSs showed the higher expression in S. spontaneum and presented expressional preference between stem (SPSA) and leaf (SPSB) tissue, speculating they might be involved in the differentia of carbohydrate metabolism in these two Saccharum species, which required further verification from experiments. Conclusions SPSA and SPSB genes presented relatively high expression and differential expression patterns between the two Saccharum species, indicating these two SPSs are important in the formation of regulatory networks and sucrose traits in the two Saccharum species. SPSB was suggested to be a major contributor to the sugar accumulation because it presented the highest expressional level and its expression positively correlated with sugar content. The recently duplicated SPSD2 presented divergent expression levels between the two Saccharum species and the relative protein content levels were highest in stem, supporting the neofunctionalization of the SPSD subfamily in Saccharum.

scholarly journals Overexpression of Sucrose Phosphate Synthase Enhanced Sucrose Content and Biomass Production in Transgenic Sugarcane

Plants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 200 ◽  
Author(s):  
Risky Mulana Anur ◽  
Nurul Mufithah ◽  
Widhi Dyah Sawitri ◽  
Hitoshi Sakakibara ◽  
Bambang Sugiharto
Keyword(s):  
Sugar Content ◽  
Sucrose Phosphate Synthase ◽  
Sucrose Content ◽  
Transgenic Sugarcane ◽  
Soluble Acid Invertase ◽  
Sugarcane Yield ◽  
Transgenic Lines ◽  
Key Enzyme ◽  
Sucrose Phosphate ◽  
Phosphate Synthase

Sucrose phosphate synthase (SPS) is a key enzyme in sucrose synthesis, which controls sucrose content in plants. This study was designed to examine the efficacy of the overexpression of SoSPS1 gene on sucrose accumulation and carbon partitioning in transgenic sugarcane. The overexpression of SoSPS1 gene increased SPS activity and sucrose content in transgenic sugarcane leaves. More importantly, the overexpression enhanced soluble acid invertase (SAI) activity concomitant with the increase of glucose and fructose levels in the leaves, whereas sucrose synthase activity exhibited almost no change. In the stalk, a similar correlation was observed, but a higher correlation was noted between SPS activity and sugar content. These results suggest that SPS overexpression has both direct and indirect effects on sugar concentration and SAI activity in sugarcane. In addition, SPS overexpression resulted in a significant increase in plant height and stalk number in some transgenic lines compared to those in non-transgenic control. Taken together, these results strongly suggest that enhancing SPS activity is a useful strategy for improving sugarcane yield.

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