Engineering α-amylase levels in wheat grain suggests a highly sophisticated level of carbohydrate regulation during development

Whan, Alex; Dielen, Anne-Sophie; Mieog, Jos C.; Bowerman, Andrew F; Robinson, Hannah; Byrne, Keren; Colgrave, Michelle L.; Larkin, P. J.; Howitt, Crispin A.; Morell, Matthew K.; Ral, Jean‐Philippe

doi:10.1093/jxb/eru299

Cited by 45 publications

(64 citation statements)

References 57 publications

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“…RNA was extracted using the protocol described in Whan et al [17]. For developing seeds, RNA was extracted from frozen halfseeds with the embryo section removed.…”

Section: Sequencing Dna and Rna Extractions And Pcr Analysesmentioning

confidence: 99%

“…TaAMY3 primers as well as primers for a reference housekeeping gene (actin) were already available [17]. Annealing sites of the four isoform-specific α-amylase primers pairs are shown in Figure S1.…”

Section: Sequencing Dna and Rna Extractions And Pcr Analysesmentioning

confidence: 99%

“…This enzyme indeed mainly appears during grain development in the pericarp, similarly to TaAMY2. Western-blot analysis suggested TaAMY3 is the predominant α-amylase throughout grain development [17]. However, its role during the grain development process remains unclear, as over-expression of this particular TaAMY3 in developing wheat endosperm showed a very limited impact on starch granule integrity despite elevated levels of α-amylase activity.…”

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confidence: 99%

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New insight in cereal starch degradation: identification and structural characterization of four α-amylases in bread wheat

Mieog

Janeček

Ral

2017

Amylase

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Grain α-amylase presents an apparent paradox for the wheat community. Despite the necessity of α-amylase for the seed germination process, high levels of amylase activity in the grain are considered detrimental for grain quality. Wheat α-amylases (EC 3.2.1.1) are endohydrolases belonging to the GH13_6 subfamily, one of the most studied subclasses of glycoside hydrolase (GH) family GH13. However, no comprehensive study had been done so far to describe and catalogue all the wheat α-amylase isoforms, despite compelling information on the involvement of two α-amylases on economically important issues for the international cereal community, namely pre-harvest sprouting and late maturity α-amylase. This study describes for the first time the genomic localization, nucleotide and amino acid sequences, phylogeny and expression profile of all known α-amylases in wheat, including a hitherto unknown fourth isoform here designated as TaAMY4. Isoform profiling strongly suggested α-amylases to be working in partnership to achieve complete degradation of a starch granule, whereas expression profiling revealed a potential involvement of TaAMY4 in the late maturity α-amylase problem.

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“…RNA was extracted using the protocol described in Whan et al [17]. For developing seeds, RNA was extracted from frozen halfseeds with the embryo section removed.…”

Section: Sequencing Dna and Rna Extractions And Pcr Analysesmentioning

confidence: 99%

Section: Sequencing Dna and Rna Extractions And Pcr Analysesmentioning

confidence: 99%

mentioning

confidence: 99%

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New insight in cereal starch degradation: identification and structural characterization of four α-amylases in bread wheat

Mieog

Janeček

Ral

2017

Amylase

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“…Деградация крахмала происходит в результате синергизма различных амилолитических ферментов (Whan et al, 2014), среди которых наиболее важную роль играет альфа-амилаза (EC 3.2.1.1, α-1,4-glucanohydro-glucanohydrolase). Идентифицирован ряд изоформ альфа-амилазы ).…”

Section: факторы стабильности высокого числа паденияunclassified

Approaches to improve wheat grain quality: breeding for falling number

Krupnov¹,

Krupnova²

2015

Vestn. VOGiS

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Agricultural research institute for South-East regions�� Saratov�� russiaFalling number (Fn� reflects the level of al�ha amylase activity in grain. The main factor res�onsible for high α amylase levels and low Fn values is visible or hidden �reharvest s�routing (PHS� under adverse weather conditions. in recent decades�� great �rogress has been made in the understanding of biochemical and molecular �rocesses accom�anying PHS and the genetic control of �reharvest s�routing resistance (PHSr� and Fn�� as well as in the develo�ment of methods of breeding for PHSr and Fn. However�� breeding for these traits and the corres�onding theoretical studies have not yet been �ro�erly develo�ed or reflected in the russian literature. The �ur�ose of the review is to fill this ga�. it illustrates the genetic com�lexity and context de�endence of the PHSr and Fn traits and of major factors im�airing Fn. A s�ecific feature of breeding for high PHSr and Fn is that it involves elimination of late maturity α-amylase (LMA� genoty�es in choosing �arents for crossing and from early hybrid generations; choice of donors of target traits with regard to their genetic diversity; methods of evaluation of varieties and lines for PHSr and Fn under different environmental conditions; determination of F 2 �o�ulation size with regard to combination of PHSr�� Fn�� high �erformance�� grain quality�� and other commercial traits. new methods and a��roaches in breeding o�en new �ros�ects. They include the doubled ha�loid (DH� method�� allowing homozygotes to be obtained from early hybrid generations; and DnA technologies�� which �ermit genetically reasonable selection of PHSr and Fn donors�� yramiding of desirable alleles�� and efficient selection of desired offs�ring using closely lin�ed molecular mar�ers.Key words�� al�ha amylase; �reharvest s�routing resistance; falling number; molecular mar�ers.Число падения (Ч�� отражает уровен�� активности ал��фа-амилазы в зерне/муке и является международным стандартом ка�ества зерна. Чем ниже Ч�� тем ниже ка�ество зерна и его цена. Главные при�ины снижения Ч� -видимое или скрытое преду�оро�ное прорастание (�� в условиях не�лагоприятной погоды. Реша�щу� рол�� в полу�ении ста�ил��но высокого Ч� играет селекция на устой�ивост�� к преду�оро�ному прорастани� (ук�� а также непосредственно на Ч�. В последние десятилетия достигнут �ол��шой прогресс в понимании �иохими�еских и молекулярных процессов�� происходящих при ��; изу�ении генети�еского контроля ук�� и Ч�� в разра�отке методов селекции на ук�� и Ч�. В России селекция на эти признаки и соответству�щие теорети�еские исследования еще не полу�или должного развития и отражения в литературе. �оказаны генети�еская сложност�� и контекст-зависимост�� признаков ук�� и Ч�. Специфика селекции на высокий уровен�� ук�� и Ч� вкл��ает удаление LMA-генотипов (LMA�� late maturity α-amylase� при под�оре родителей для скрещивания и в ранних поколениях ги�ридов�� вы�ор генети�ески разли�ных доноров целевых признаков�� методы оценки сортов и линий на ук�� и Ч� в разных условиях в...

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“…In wheat, starch makes up over 70% of total grain dry weight and provides the energy required by the embryo to fuel metabolic processes during the first days of grain germination [6,7]. Therefore, starch has been considered one of the primary targets for agronomic performance enhancement.…”

Section: Introductionmentioning

confidence: 99%

Transferring a Biomass Enhancement Biotechnology from Glasshouse to Field: A Case Study on Wheat GWD RNAi

et al. 2017

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Abstract:In glasshouse studies we have previously shown that endosperm-specific RNAi suppression of the primary starch phosphorylation enzyme, Glucan, Water Dikinase (GWD) leads to enhanced early vigor, greater leaf biomass, and increases in both head size and yield. To confirm these affects in a field setting, trials were conducted in three Australian environments. Field results were consistent with those in the glasshouse for increased flag leaf area and rachis nodes. However, there was also a decrease in tiller number and consequently a decrease in yield for one event at two sites. These findings provide potentially important information on plant vigor enhancement and highlight the challenges of transferring the modification of complex traits from single plants in controlled environments to the field.

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Engineering α-amylase levels in wheat grain suggests a highly sophisticated level of carbohydrate regulation during development

Abstract: SummaryEndosperm-specific over-expression of wheat α-amylase TaAMY3 during grain development revealed unexpected effects on grain composition, carbon partitioning, fatty acid accumulation, and germination.

Cited by 45 publications

References 57 publications

New insight in cereal starch degradation: identification and structural characterization of four α-amylases in bread wheat

New insight in cereal starch degradation: identification and structural characterization of four α-amylases in bread wheat

Approaches to improve wheat grain quality: breeding for falling number

Transferring a Biomass Enhancement Biotechnology from Glasshouse to Field: A Case Study on Wheat GWD RNAi

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