Development of Allergenicity and Toxicity Assessment Methods for Evaluating Transgenic Sugarcane Overexpressing Sucrose–Phosphate Synthase

Neliana, Intan Ria; Sawitri, Widhi Dyah; Ermawati, Netty; Handoyo, Tri; Sugiharto, Bambang

doi:10.3390/agronomy9010023

Cited by 3 publications

(2 citation statements)

References 43 publications

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“…Contrarily, sucrose synthase activities and their gene expression are substantially higher in young sugarcane internodes, possibly providing carbon for cellulose and cell walls synthesized from uridine-diphosphate-glucose (UDP-G) (Mason et al, 2023). A substantial decrease in SPS-specific activity was exhibited in both varieties, suggesting adverse effects on enzyme functions in sugarcane cultivars due to heat shock treatments, which were also reported in another genotype of sugarcane (Neliana et al, 2019).This finding suggested that sugarcane plants demand optimum temperatures for survival, especially for maximum sucrose accumulation in the stem at the maturity stage. Maximum SPS expression was exhibited at the maturity stage and the correlation analysis showed that the sucrose content had a strongly positive association with SPS.…”

Section: Discussionmentioning

confidence: 75%

Expression of sucrose metabolizing enzymes in different sugarcane varieties under progressive heat stress

Mehdi,

Liu,

Riaz

et al. 2023

Front. Plant Sci.

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Studying the thermal stress effect on sucrose-metabolizing enzymes in sugarcane is of great importance for understanding acclimation to thermal stress. In this study, two varieties, S2003-US-633 and SPF-238, were grown at three different temperatures ( ± 2°C): 30°C as a control, 45°C for various episodes of high temperature treatments and recovery conditions at 24, 48 and 72 hours. Data showed that reducing sugar content increased until the grand growth stage but sharply declined at the maturity stage in both cultivars. On the other hand, sucrose is enhanced only at the maturity stage. The expression of all invertase isozymes declined prominently; however, the expression of SPS was high at the maturity stage. Hence, the sucrose accumulation in mature cane was due to increased SPS activity while decreased invertase isozymes (vacuolar, cytoplasmic and cell wall) activities at maturity stage in both cultivars. Heat shock decreased the sucrose metabolizing enzymes, sucrose content and sugar recovery rate in both cultivars. In contrast, heat-shock treatments induced maximum proline, MDA, H2O2 and EC in both cultivars. Notably, this is the first report of diverse invertase isozyme molecular weight proteins, such as those with 67, 134 and 160 kDa, produced under heat stress, suggesting that these enzymes have varied activities at different developmental stages. Overall, S2003-US-633 performs better than the cultivar SPF-238 under heat stress conditions at all development stages, with increased sucrose content, enzyme expression, proline and sugar recovery rate. This work will provide a new avenue regarding sugarcane molecular breeding programs with respect to thermal stress.

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Section: Discussionmentioning

confidence: 75%

Expression of sucrose metabolizing enzymes in different sugarcane varieties under progressive heat stress

Mehdi,

Liu,

Riaz

et al. 2023

Front. Plant Sci.

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“…Enzim yang digunakan dalam uji aktifitas enzim didapatkan dari daun transgenik tomat. Metode ekstraksi mengacu pada Torres et al 1989 dengan sedikit modifikasi (Sawitri et al 2016;Neliana et al 2019). Daun tomat digerus dengan nitrogen cair dan dicampur dengan modifikasi buffer ekstraksi sebanyak 3 kali volum [50mM Tris-HCl (pH 7,5), 150mM NaCl, 1mM MgCl2, 1mM EDTA, 2,5mM DTT dan 10% PVPP].…”

Section: Ekspresi Protein Melalui Aktifitas Enzimunclassified

TRANSFORMASI GENETIK DAN EKSPRESI MUTAN <em>SUCROSE PHOSPHATE SYNTHASE</em> PADA TANAMAN TOMAT

Fibriani

Agustien

Sawitri

et al. 2019

J Bioteknol Biosains Indones

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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

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Enhancing sugarcane growth quality and productivity through a biotechnology approach

Sugiharto

2024

BIO Web Conf.

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Sugarcane is a major crop to produce sugar accounting for nearly 80% of sugar production worldwide. Sugarcane is well adapted to warm climates and accumulated high biomass quantities for bioelectricity and second-generation bioethanol. Although Indonesia is one among the cane sugar producers, the produced sugar has been unable to meet the national sugar demand. The study of physiology, molecular biology and genetic is providing a major impetus to develop biotechnological strategies for increasing growth and productivity in sugarcane. Genetic transformation method for sugarcane has been established, including Agrobacterium-mediated transformation method. The Agrobacterium-mediated transformation has been successfully employed to develop transgenic sugarcane. The overexpression of SoSPS gene encoding for sucrosephosphate synthase (SPS) showed the increases of activity and sucrose content in transgenic sugarcane. Furthermore, field evaluation on growth and productivity of the transgenic sugarcane displayed higher tiller number, plant high, cane yield, percentage of Brix and Pol compared to nontransgenic sugarcane. Furthermore, plants are subjected to a variety of abiotic and biotic stresses, which reduces and limits crop productivity. Plants adapt to water stress with various strategies include change in the gene expression and accumulation of organic compounds called compatible solutes. Genetic transformation of betA gene encoding for choline dehydrogenase in bacteria elevated glycine-betaine content as an osmoprotectant and resulted in water stress tolerant of transgenic sugarcane. The drought tolerant of sugarcane was already approved and released by Indonesian Government for commercialization. In addition, mosaic virus is one of the most severe diseases in sugarcane and lead to the constant losses in growth and yield of sugarcane. Pathogen-derived resistance (PDR) and RNA interference (RNAi) technologies have been applied to engineered sugarcane cultivar resistant to mosaic virus, and that the RNAi method produced more resistant against the mosaic virus in sugarcane. Finaly, biotechnology approach of genome editing technology should be exploited to ensure higher sugarcane productivity, and improve the livelihoods of smallholder farmers.

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Development of Allergenicity and Toxicity Assessment Methods for Evaluating Transgenic Sugarcane Overexpressing Sucrose–Phosphate Synthase

Cited by 3 publications

References 43 publications

Expression of sucrose metabolizing enzymes in different sugarcane varieties under progressive heat stress

Expression of sucrose metabolizing enzymes in different sugarcane varieties under progressive heat stress

TRANSFORMASI GENETIK DAN EKSPRESI MUTAN <em>SUCROSE PHOSPHATE SYNTHASE</em> PADA TANAMAN TOMAT

Enhancing sugarcane growth quality and productivity through a biotechnology approach

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