Post-translational regulation of acid invertase activity by vacuolar invertase inhibitor affects resistance to cold-induced sweetening of potato tubers

2012 ◽  
Vol 36 (1) ◽  
pp. 176-185 ◽  
Author(s):  
MARIAN J. MCKENZIE ◽  
RONAN K. Y. CHEN ◽  
JOHN C. HARRIS ◽  
MATTHEW J. ASHWORTH ◽  
DAVID A. BRUMMELL
Keyword(s):  
Translational Regulation ◽  
Invertase Activity ◽  
Acid Invertase ◽  
Potato Tubers ◽  
Vacuolar Invertase ◽  
Invertase Inhibitor ◽  
Cold Induced Sweetening

scholarly journals Induction of vacuolar invertase inhibitor mRNA in potato tubers contributes to cold-induced sweetening resistance and includes spliced hybrid mRNA variants

2011 ◽  
Vol 62 (10) ◽  
pp. 3519-3534 ◽  
Author(s):  
David A. Brummell ◽  
Ronan K. Y. Chen ◽  
John C. Harris ◽  
Huaibi Zhang ◽  
Cyril Hamiaux ◽  
...  
Keyword(s):  
Potato Tubers ◽  
Vacuolar Invertase ◽  
Invertase Inhibitor ◽  
Mrna Variants ◽  
Cold Induced Sweetening

scholarly journals StRAP2.3, an ERF‐VII transcription factor, directly activates StInvInh2 to enhance cold-induced sweetening resistance in potato

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Weiling Shi ◽  
Yuhao Song ◽  
Tiantian Liu ◽  
Qiuqin Ma ◽  
Wang Yin ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Promoter Region ◽  
Expression Patterns ◽  
Potato Tubers ◽  
Vacuolar Invertase ◽  
Cis Element ◽  
Invertase Inhibitor ◽  
Cold Induced Sweetening ◽  
In Cis

AbstractPotato invertase inhibitor (StInvInh2) positively regulates cold-induced sweetening (CIS) resistance by inhibiting the activity of vacuolar invertase. The distinct expression patterns of StInvInh2 have been thoroughly characterized in different potato genotypes, but the related CIS ability has not been characterized. The understanding of the regulatory mechanisms that control StInvInh2 transcription is unclear. In this study, we identified an ERF‐VII transcription factor, StRAP2.3, that directly regulates StInvInh2 to positively modulate CIS resistance. Acting as a nuclear-localized transcriptional activator, StRAP2.3 directly binds the ACCGAC cis-element in the promoter region of StInvInh2, enabling promoter activity. Overexpression of StRAP2.3 in CIS-sensitive potato tubers induced StInvInh2 mRNA abundance and increased CIS resistance. In contrast, silencing StRAP2.3 in CIS-resistant potato tubers repressed the expression of StInvInh2 and decreased CIS resistance. We conclude that cold-responsive StInvInh2 is due to the binding of StRAP2.3 to the ACCGAC cis-element in the promoter region of StInvInh2. Overall, these findings indicate that StRAP2.3 directly regulates StInvInh2 to positively modulate CIS resistance, which may provide a strategy to improve the processing quality of potatoes.

Разработка и тестирование универсальных праймеров для ПЦР-амплификации генов-ортологов β-фруктофуранозидазы (Pain-1) у видов и сортов картофеля

2018 ◽  
Author(s):  
E.O. Shmelkova ◽  
M.A. Slugina ◽  
A.A. Meleshin ◽  
E.V. Romanova
Keyword(s):  
Solanum Tuberosum ◽  
Wild Species ◽  
Starch Synthesis ◽  
Full Length ◽  
Universal Primers ◽  
Potato Tubers ◽  
Vacuolar Invertase ◽  
Reserve Carbohydrate ◽  
Cold Induced Sweetening ◽  
Design And Testing

Работа посвящена разработке и тестированию универсальных праймеров для ПЦР-амплификации полноразмерных генов-ортологов β-фруктофуранозидазы (кислой вакуолярной инвертазы) у видов и сортов картофеля (Solanum tuberosum). Крахмал – основной источник энергии и резервный углевод, накапливающийся в амилопластах клубней. Образовавшаяся в результате фотосинтеза молекула глюкозы при реакции с фруктозой образует сахарозу – основную транспортную форму углеводов в растении. В клубни сахароза доставляется по флоэме (апопластный путь), где в межклеточном пространстве расщепляется до глюкозы и фруктозы, которые затем проникают в клетки паренхимы. Глюкоза служит в дальнейшем субстратом для синтеза крахмала в амилопластах. Однако при воздействии пониженных температур крахмал в клубнях картофеля разрушается до редуцирующих сахаров. Параллельно этому процессу идет ресинтез сахарозы до глюкозы и фруктозы за счет фермента кислой вакуолярной инвертазы (β-фруктофуранозидазы), кодируемой геном Pain-1. В совокупности эти процессы приводят к избыточному накоплению моносахаров в клубнях картофеля, так называемому холодовому осахариванию (cold-induced sweetening). При этом создаются условия для интенсивного образования меланоидинов, вызывающих потемнение мякоти картофеля, что значительно ухудшает товарное качество продукта. Таким образом, изучение гена Pain-1, кодирующего вакуолярную инвертазу, а именно, его идентификация и анализ структуры – важная задача, необходимая для поиска доноров, устойчивых к холодовому осахариванию. Первоочередная задача для этого – разработка и тестирование праймерных комбинаций, позволяющих амплифицировать полноразмерный ген у диких видов картофеля, а также сортов и линий культивируемого картофеля (S. tuberosum). В данной работе приведены результаты разработки и тестирования универсальных праймеров, с помощью которых можно амплифицировать как полноразмерные гены-ортологи, так и фрагменты гена Pain-1, а также подобраны оптимальные условия для проведения ПЦР реакции. Было разработано 6 праймерных комбинаций (PainF – PainR, PainF – Pain1exR, Pain1exF – Pain3exR, Pain2inF – Pain2inR, Pain3exF – Pain5exR, Pain5exF – PainR), среди которых комбинация PainF – PainR позволяла амплифицировать полноразмерный ген, остальные – внутренние и будут использованы в дальнейшем при секвенировании фрагментов исследуемого гена. Эти праймеры были успешно протестированы на 15 образцах, включающих представителей пяти дикорастущих видов картофеля (S. gourlay, S. chacoense, S. pinnatissectum, S. stoloniferum, S. vernei) и десяти сортов российской и зарубежной селекции (Гала, Ласунок, Ред Скарлетт, Рассет Бербанк, Мирас, Башкирский, Жуковский ранний, Матушка, Елизавета, Сударыня).The purpose of research is design and testing of universal primers for PCR amplification of full-length-fructofuranozidase orthologs genes (acid vacuolar invertase) in wild species and potato (Solanum tuberosum) varieties. Starch is the main source of energy and a reserve carbohydrate, that accumulates in tubers amyloplasts. Glucose molecule, produced by photosynthesis, reacts with fructose and forms sucrose, which is the main transport type of carbohydrates in the plant. In the tuber, sucrose is delivered via phloem (apoplast), where it splits into glucose and fructose, which then go to the parenchyma cells. Glucose is a further substrate for the starch synthesis in amyloplasts. However, low temperatures influence on potato tubers leads to starch break down to reducing sugars. In parallel to this process there is happens resynthesis of sucrose to glucose and fructose by acid vacuolar invertase enzyme (β-fructofuranosidase) encoded by Pain-1 gene. Together, these processes lead to an excessive accumulation of monosaccharides in potato tubers. This process also called as cold-induced sweetening. It creates conditions for the intensive formation of melanoidins, which cause a potato tubers darkening, which considerably impairs the commercial quality of the product. Thus, the study Pain-1 gene that encodes the vacuolar invertase (its identification and structure analysis) is an important task required for the search of donors resistant to cold-induced sweetening. The primary task for this is the design and testing of primer combinations that allow to amplify the full-length gene in wild potato species, varieties and lines of cultivated potato. In this work, we develop and test universal primers, that can amplify both full-length orthologs and fragments of the Pain-1 gene, and also select the optimal conditions for carrying out the PCR reaction. Summary. The purpose of research is design and testing of universal primers for PCR amplification of full-length-fructofuranozidase orthologs genes (acid vacuolar invertase) in wild species and potato (Solanum tuberosum) varieties. Starch is the main source of energy and a reserve carbohydrate, that accumulates in tubers amyloplasts. Glucose molecule, produced by photosynthesis, reacts with fructose and forms sucrose, which is the main transport type of carbohydrates in the plant. In the tuber, sucrose is delivered via phloem (apoplast), where it splits into glucose and fructose, which then go to the parenchyma cells. Glucose is a further substrate for the starch synthesis in amyloplasts. However, low temperatures influence on potato tubers leads to starch break down to reducing sugars. In parallel to this process there is happens resynthesis of sucrose to glucose and fructose by acid vacuolar invertase enzyme (β-fructofuranosidase) encoded by Pain-1 gene. Together, these processes lead to an excessive accumulation of monosaccharides in potato tubers. This process also called as cold-induced sweetening. It creates conditions for the intensive formation of melanoidins, which cause a potato tubers darkening, which considerably impairs the commercial quality of the product. Thus, the study Pain-1 gene that encodes the vacuolar invertase (its identification and structure analysis) is an important task required for the search of donors resistant to cold-induced sweetening. The primary task for this is the design and testing of primer combinations that allow to amplify the full-length gene in wild potato species, varieties and lines of cultivated potato. In this work, we develop and test universal primers, that can amplify both full-length orthologs and fragments of the Pain-1 gene, and also select the optimal conditions for carrying out the PCR reaction. In total 6 primer combinations were designed (PainF - PainR, PainF - Pain1exR, Pain1exF - Pain3exR, Pain2inF - Pain2inR, Pain3exF - Pain5exR, Pain5exF - PainR), where PainF - PainR primer combination allowed to amplify a full-sized gene, the rest are internal and will be used in the further fragments sequencing of the β-fructofuranosidase gene. These primers were successfully tested on 15 samples, including five wild species of potato (S. gourlay, S. chacoense, S. pinnatissectum, S. stoloniferum, S. vernei) and ten varieties of Russian and foreign breeding (Gala, Lasunok, Red Scarlet , Rasset Burbank, Miras, Bashkirsky, Zhukovsky ranniy, Matushka, Elizaveta, Sudaryna).

Vacuolar sucrose homeostasis is critical for plant development, seed properties, and night-time survival in Arabidopsis

2020 ◽  
Vol 71 (16) ◽  
pp. 4930-4943 ◽  
Author(s):  
Duc Phuong Vu ◽  
Cristina Martins Rodrigues ◽  
Benjamin Jung ◽  
Garvin Meissner ◽  
Patrick A W Klemens ◽  
...  
Keyword(s):  
Seed Weight ◽  
Plant Cells ◽  
Invertase Activity ◽  
Frost Tolerance ◽  
Wild Type ◽  
Leaf Extracts ◽  
Night Time ◽  
Vacuolar Invertase ◽  
Cold Temperatures ◽  
Invertase Inhibitor

Abstract Most cellular sucrose is present in the cytosol and vacuoles of plant cells; however, little is known about the effect of this sucrose compartmentation on plant properties. Here, we examined the effects of altered intracellular sucrose compartmentation in Arabidopsis thaliana leaves by heterologously expressing the sugar beet (Beta vulgaris) vacuolar sucrose loader BvTST2.1 and by generating lines with reduced vacuolar invertase activity (amiR vi1-2). Heterologous expression of BvTST2.1 led to increased monosaccharide levels in leaves, whereas sucrose levels remained constant, indicating that vacuolar invertase activity in mesophyll vacuoles exceeds sucrose uptake. This notion was supported by analysis of tobacco (Nicotiana benthamiana) leaves transiently expressing BvTST2.1 and the invertase inhibitor NbVIF. However, sucrose levels were strongly elevated in leaf extracts from amiR vi1-2 lines, and experiments confirmed that sucrose accumulated in the corresponding vacuoles. The amiR vi1-2 lines exhibited impaired early development and reduced seed weight. When germinated in the dark, amiR vi1-2 seedlings were less able to convert sucrose into monosaccharides than the wild type. Cold temperatures strongly down-regulated both VI genes, but the amiR vi1-2 lines showed normal frost tolerance. These observations indicate that increased vacuolar sucrose levels fully compensate for the effects of low monosaccharide concentrations on frost tolerance.

Ectopic expression of a tobacco invertase inhibitor homolog prevents cold-induced sweetening of potato tubers

10.1038/10924 ◽  
1999 ◽  
Vol 17 (7) ◽  
pp. 708-711 ◽  
Author(s):  
Steffen Greiner ◽  
Thomas Rausch ◽  
Uwe Sonnewald ◽  
Karin Herbers
Keyword(s):  
Ectopic Expression ◽  
Potato Tubers ◽  
Invertase Inhibitor ◽  
Cold Induced Sweetening
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