scholarly journals Transcriptome analysis of genes involved in starch biosynthesis in developing Chinese chestnut (Castanea mollissima Blume) seed kernels

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Lingling Shi ◽  
Jia Wang ◽  
Yujun Liu ◽  
Chao Ma ◽  
Sujuan Guo ◽  
...  
Keyword(s):  
Regulatory Mechanism ◽  
Average Length ◽  
Starch Content ◽  
Raw Material ◽  
Starch Accumulation ◽  
Potential Candidate ◽  
Sequencing Analysis ◽  
Chinese Chestnut ◽  
Castanea Mollissima ◽  
Genes Encoding

AbstractChinese chestnut (Castanea mollissima Blume) seed kernels (CCSK) with high quality and quantity of starch has emerged as a potential raw material for food industry, but the molecular regulatory mechanism of starch accumulation in developing CCSK is still unclear. In this study, we firstly analyzed the fruit development, starch accumulation, and microscopic observation of dynamic accumulation of starch granules of developing CCSK from 10 days after flowering (DAF) to 100 DAF, of which six representative CCSK samples (50–100 DAF) were selected for transcriptome sequencing analysis. Approximately 40 million valid reads were obtained, with an average length of 124.95 bp, which were searched against a reference genome, returning 38,146 unigenes (mean size = 1164.19 bp). Using the DESeq method, 1968, 1573, 1187, 1274, and 1494 differentially expressed unigenes were identified at 60:50, 70:60, 80:70, 90:80 and 100:90 DAF, respectively. The relationship between the unigene transcriptional profiles and starch dynamic patterns in developing CCSK was comparatively analyzed, and the specific unigenes encoding for metabolic enzymes (SUSY2, PGM, PGI, GPT, NTT, AGP3, AGP2, GBSS1, SS1, SBE1, SBE2.1, SBE2.2, ISA1, ISA2, ISA3, and PHO) were characterized to be involved potentially in the biosynthesis of G-1-P, ADPG, and starch. Finally, the temporal transcript profiles of genes encoding key enzymes (susy2, pgi2, gpt1, agp2, agp3, gbss1, ss1, sbe1, sbe2.1, sbe2.2, isa1, isa2, isa3, and pho) were validated by quantitative real-time PCR (qRT-PCR). Our findings could help to reveal the molecular regulatory mechanism of starch accumulation in developing CCSK and may also provide potential candidate genes for increasing starch content in Chinese chestnut or other starchy crops.

scholarly journals Transcriptome Analysis and Identification of Genes Associated with Starch Metabolism in Castanea henryi Seed (Fagaceae)

2020 ◽  
Vol 21 (4) ◽  
pp. 1431 ◽  
Author(s):  
Bin Liu ◽  
Ruqiang Lin ◽  
Yuting Jiang ◽  
Shuzhen Jiang ◽  
Yuanfang Xiong ◽  
...  
Keyword(s):  
Seed Germination ◽  
Candidate Genes ◽  
Starch Content ◽  
Sugar Content ◽  
Soluble Sugar ◽  
Starch Accumulation ◽  
Transcriptional Level ◽  
Potential Candidate ◽  
Starch Metabolism ◽  
Carbohydrate Storage

Starch is the most important form of carbohydrate storage and is the major energy reserve in some seeds, especially Castanea henryi. Seed germination is the beginning of the plant’s life cycle, and starch metabolism is important for seed germination. As a complex metabolic pathway, the regulation of starch metabolism in C. henryi is still poorly understood. To explore the mechanism of starch metabolism during the germination of C. henryi, we conducted a comparative gene expression analysis at the transcriptional level using RNA-seq across four different germination stages, and analyzed the changes in the starch and soluble sugar contents. The results showed that the starch content increased in 0–10 days and decreased in 10–35 days, while the soluble sugar content continuously decreased in 0–30 days and increased in 30–35 days. We identified 49 candidate genes that may be associated with starch and sucrose metabolism. Three ADP-glucose pyrophosphorylase (AGPase) genes, two nucleotide pyrophosphatase/phosphodiesterases (NPPS) genes and three starch synthases (SS) genes may be related to starch accumulation. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to validate the expression levels of these genes. Our study combined transcriptome data with physiological and biochemical data, revealing potential candidate genes that affect starch metabolism during seed germination, and provides important data about starch metabolism and seed germination in seed plants.

scholarly journals Transcriptome Analysis of Chinese Chestnut (Castanea mollissima Blume) in Response to Dryocosmus kuriphilus Yasumatsu Infestation

2019 ◽  
Vol 20 (4) ◽  
pp. 855 ◽  
Author(s):  
Cancan Zhu ◽  
Fenghou Shi ◽  
Yu Chen ◽  
Min Wang ◽  
Yuqiang Zhao ◽  
...  
Keyword(s):  
Growth Stage ◽  
Differentially Expressed ◽  
Maturation Stage ◽  
Gall Formation ◽  
Chinese Chestnut ◽  
Dryocosmus Kuriphilus ◽  
Castanea Mollissima ◽  
Genes Encoding ◽  
Phenylpropanoid Biosynthesis ◽  
Initiation Stage

Chinese chestnut (Castanea mollissima Blume) can be infested by Dryocosmus kuriphilus Yasumatsu, resulting in gall formation and yield losses. Research on the control of gall wasps using genomics approaches is rarely reported. We used RNA-seq to investigate the dynamic changes in the genes of a chestnut species (C. mollissima B.) during four gall-formation stages caused by D. kuriphilus. A total of 21,306 genes were annotated by BLAST in databases. Transcriptome comparison between different gall-formation stages revealed many genes that were differentially expressed compared to the control. Among these, 2410, 7373, 6294, and 9412 genes were differentially expressed in four gall-formation stages: initiation stage (A), early growth stage (B), late growth stage (C), and maturation stage (D), respectively. Annotation analysis indicated that many metabolic processes (e.g., phenylpropanoid biosynthesis, secondary metabolism, plant–pathogen interaction) were affected. Interesting genes encoding putative components of signal transduction, stress response, and transcription factors were also differentially regulated. These genes might play important roles in response to D. kuriphilus gall formation. These new data on the mechanism by which D. kuriphilus infests chestnuts could help improve chestnut resistance.

Co-expression of AGPS and AGPL genes encoded for AGPase from cassava to enhance starch accumulation in transgenic tobacco

2016 ◽  
Vol 14 (2) ◽  
pp. 287-293
Author(s):  
Nguyễn Văn Đoài ◽  
Nguyễn Minh Hồng ◽  
Lê Thu Ngọc ◽  
Nguyễn Thị Thơm ◽  
Nguyễn Đình Trọng ◽  
...  
Keyword(s):  
Transgenic Tobacco ◽  
Higher Plants ◽  
Starch Content ◽  
Genetically Modified Crops ◽  
Starch Accumulation ◽  
35S Promoter ◽  
Effective Strategies ◽  
Plant Expression Vector ◽  
Transgenic Lines ◽  
Cassava Varieties

The AGPase (ADP-Glucose pyrophosphorylase) is one of the ubiquitous enzymes catalyzing the first step in starch biosynthesis. It plays an important role in regulation and adjusts the speed of the entire cycle of glycogen biosynthesis in bacteria and starch in plants. In higher plants, it is a heterotetramer and tetrameric enzyme consisting two large subunits (AGPL) and two small subunits (AGPS) and encoded by two genes. In this paper, both AGPS and AGPL genes were sucessfully isolated from cassava varieties KM140 and deposited in Genbank with accession numbers KU243124 (AGPS) and KU243122 (AGPL), these two genes were fused with P2a and inserted into plant expression vector pBI121 under the control of 35S promoter. The efficient of this construct was tested in transgenic N. tabacum. The presence and expression of AGPS and AGPL in transgenic plants were confirmed by PCR and Western hybridization. The starch content was quantified by the Anthrone method. Transgenic plant analysis indicated that that two targeted genes were expressed simultaneously in several transgenic tobacco lines under the control of CaMV 35S promoter.  The starch contents in 4 analyzed tobacco transgenic lines displays the increase 13-116%  compared to WT plants. These results indicated that the co-expression of AGPS and AGPL is one of effective strategies for enhanced starch production in plant. These results can provide a foundation for developing other genetically modified crops to increase starch accumulation capacity.

scholarly journals β-Galactosidases from a Sequence-Based Metagenome: Cloning, Expression, Purification and Characterization

2020 ◽  
Vol 9 (1) ◽  
pp. 55
Author(s):  
María Florencia Eberhardt ◽  
José Matías Irazoqui ◽  
Ariel Fernando Amadio
Keyword(s):  
Bacterial Species ◽  
Value Added ◽  
Raw Material ◽  
Activity Levels ◽  
Treatment Technology ◽  
Catalytic Domains ◽  
Stabilization Ponds ◽  
Cazy Database ◽  
Genes Encoding ◽  
Value Added Products

Stabilization ponds are a common treatment technology for wastewater generated by dairy industries. Large proportions of cheese whey are thrown into these ponds, creating an environmental problem because of the large volume produced and the high biological and chemical oxygen demands. Due to its composition, mainly lactose and proteins, it can be considered as a raw material for value-added products, through physicochemical or enzymatic treatments. β-Galactosidases (EC 3.2.1.23) are lactose modifying enzymes that can transform lactose in free monomers, glucose and galactose, or galactooligosacharides. Here, the identification of novel genes encoding β-galactosidases, identified via whole-genome shotgun sequencing of the metagenome of dairy industries stabilization ponds is reported. The genes were selected based on the conservation of catalytic domains, comparing against the CAZy database, and focusing on families with β-galactosidases activity (GH1, GH2 and GH42). A total of 394 candidate genes were found, all belonging to bacterial species. From these candidates, 12 were selected to be cloned and expressed. A total of six enzymes were expressed, and five cleaved efficiently ortho-nitrophenyl-β-galactoside and lactose. The activity levels of one of these novel β-galactosidase was higher than other enzymes reported from functional metagenomics screening and higher than the only enzyme reported from sequence-based metagenomics. A group of novel mesophilic β-galactosidases from diary stabilization ponds’ metagenomes was successfully identified, cloned and expressed. These novel enzymes provide alternatives for the production of value-added products from dairy industries’ by-products.

scholarly journals Starch Production in Chlamydomonas reinhardtii through Supraoptimal Temperature in a Pilot-Scale Photobioreactor

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1084
Author(s):  
Ivan N. Ivanov ◽  
Vilém Zachleder ◽  
Milada Vítová ◽  
Maria J. Barbosa ◽  
Kateřina Bišová
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Starch Content ◽  
Light Availability ◽  
Fold Increase ◽  
Pilot Scale ◽  
Temperature Treatment ◽  
Control Culture ◽  
Starch Accumulation ◽  
Cellular Division ◽  
Starch Production

An increase in temperature can have a profound effect on the cell cycle and cell division in green algae, whereas growth and the synthesis of energy storage compounds are less influenced. In Chlamydomonas reinhardtii, laboratory experiments have shown that exposure to a supraoptimal temperature (39 °C) causes a complete block of nuclear and cellular division accompanied by an increased accumulation of starch. In this work we explore the potential of supraoptimal temperature as a method to promote starch production in C. reinhardtii in a pilot-scale photobioreactor. The method was successfully applied and resulted in an almost 3-fold increase in the starch content of C. reinhardtii dry matter. Moreover, a maximum starch content at the supraoptimal temperature was reached within 1–2 days, compared with 5 days for the control culture at the optimal temperature (30 °C). Therefore, supraoptimal temperature treatment promotes rapid starch accumulation and suggests a viable alternative to other starch-inducing methods, such as nutrient depletion. Nevertheless, technical challenges, such as bioreactor design and light availability within the culture, still need to be dealt with.

scholarly journals Molecular mechanism underlying the effect of maleic hydrazide treatment on starch accumulation in S. polyrrhiza 7498 fronds

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yerong Zhu ◽  
Xiaoxue Li ◽  
Xuan Gao ◽  
Jiqi Sun ◽  
Xiaoyuan Ji ◽  
...  
Keyword(s):  
Plant Growth ◽  
Carbon Fixation ◽  
Maleic Hydrazide ◽  
Starch Content ◽  
Vegetative Reproduction ◽  
Photosynthetic Pigment ◽  
Starch Accumulation ◽  
Chlorophyll Fluorescence Parameters ◽  
Significant Difference ◽  
Starch Production

Abstract Background Duckweed is considered a promising feedstock for bioethanol production due to its high biomass and starch production. The starch content can be promoted by plant growth regulators after the vegetative reproduction being inhibited. Maleic hydrazide (MH) has been reported to inhibit plant growth, meantime to increase biomass and starch content in some plants. However, the molecular explanation on the mechanism of MH action is still unclear. Results To know the effect and action mode of MH on the growth and starch accumulation in Spirodela polyrrhiza 7498, the plants were treated with different concentrations of MH. Our results showed a substantial inhibition of the growth in both fronds and roots, and increase in starch contents of plants after MH treatment. And with 75 µg/mL MH treatment and on the 8th day of the experiment, starch content was the highest, about 40 mg/g fresh weight, which is about 20-fold higher than the control. The I2-KI staining and TEM results confirmed that 75 µg/mL MH-treated fronds possessed more starch and big starch granules than that of the control. No significant difference for both in the photosynthetic pigment content and the chlorophyll fluorescence parameters of PII was found. Differentially expressed transcripts were analyzed in S. polyrrhiza 7498 after 75 µg/mL MH treatment. The results showed that the expression of some genes related to auxin response reaction was down-regulated; while, expression of some genes involved in carbon fixation, C4 pathway of photosynthesis, starch biosynthesis and ABA signal transduction pathway was up-regulated. Conclusion The results provide novel insights into the underlying mechanisms of growth inhibition and starch accumulation by MH treatment, and provide a selective way for the improvement of starch production in duckweed.

scholarly journals PSB27: A thylakoid protein enabling Arabidopsis to adapt to changing light intensity

2015 ◽  
Vol 112 (5) ◽  
pp. 1613-1618 ◽  
Author(s):  
Xin Hou ◽  
Aigen Fu ◽  
Veder J. Garcia ◽  
Bob B. Buchanan ◽  
Sheng Luan
Keyword(s):  
Photosystem Ii ◽  
Light Intensity ◽  
Light Adaptation ◽  
Light Exposure ◽  
Starch Accumulation ◽  
State Transitions ◽  
Potential Candidate ◽  
Subsequent Work ◽  
Variable Intensity ◽  
Changing Light

In earlier studies we have identified FKBP20-2 and CYP38 as soluble proteins of the chloroplast thylakoid lumen that are required for the formation of photosystem II supercomplexes (PSII SCs). Subsequent work has identified another potential candidate functional in SC formation (PSB27). We have followed up on this possibility and isolated mutants defective in the PSB27 gene. In addition to lack of PSII SCs, mutant plants were severely stunted when cultivated with light of variable intensity. The stunted growth was associated with lower PSII efficiency and defective starch accumulation. In response to high light exposure, the mutant plants also displayed enhanced ROS production, leading to decreased biosynthesis of anthocyanin. Unexpectedly, we detected a second defect in the mutant, namely in CP26, an antenna protein known to be required for the formation of PSII SCs that has been linked to state transitions. Lack of PSII SCs was found to be independent of PSB27, but was due to a mutation in the previously described cp26 gene that we found had no effect on light adaptation. The present results suggest that PSII SCs, despite being required for state transitions, are not associated with acclimation to changing light intensity. Our results are consistent with the conclusion that PSB27 plays an essential role in enabling plants to adapt to fluctuating light intensity through a mechanism distinct from photosystem II supercomplexes and state transitions.

scholarly journals Mechanical properties of Gelatin –Hydroxyapatite composite for bone tissue engineering

2015 ◽  
Vol 50 (1) ◽  
pp. 15-20 ◽  
Author(s):  
MJ Hossan ◽  
MA Gafur ◽  
MM Karim ◽  
AA Rana
Keyword(s):  
Mechanical Properties ◽  
Bone Tissue ◽  
Composite Scaffold ◽  
Testing Machine ◽  
Casting Process ◽  
Raw Material ◽  
Potential Candidate ◽  
Oven Drying ◽  
Hydroxyapatite Composite ◽  
Properties Of Composites

In this study, hydroxyapatite (HAp) and gelatin (GEL) scaffolds were prepared to mimic the mineral and organic component of natural bone. The raw material was first compounded and resulting composite were molded into the petridishes. Using Solvent casting process, it is possible to produce scaffolds with mechanical and structural properties close to natural trabecular bone.The mechanical properties of composites were investigated by Thermo-mechanical analyzer (TMA), Vickers microhardness tester, Universal testing machine. It was observed that the composite has maximum tensile strength of 37.13MPa ( oven drying) and % elongation of 7.68 (Oven drying) and 2.04 (Natural drying) at 15% of Hap respectively. These results demonstrate that the prepared composite scaffold is a potential candidate for bone tissue engineering.Bangladesh J. Sci. Ind. Res. 50(1), 15-20, 2015

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