Storage Protein Genes Research Articles

Bixafen, Prothioconazole, and Trifloxystrobin Alone or in Combination Have a Greater Effect on Health Related Gene Expression in Honey Bees from Nutritionally Deprived than from Protein Supplemented Colonies.

The aim of this study was to evaluate whether alterations in food availability compromise the metabolic homeostasis of honey bees exposed to three fungicides alone or together. Ten honey bee colonies were used, with half receiving carbohydrate-protein supplementation for 15 weeks while another five colonies had their protein supply reduced with pollen traps. Subsequently, forager bees were collected and exposed by contact to 1 or 7 µg of bixafen, prothioconazole, or trifloxystrobin, either individually or in combination. After 48 h, bee abdomens without the intestine were used for the analysis of expression of antioxidant genes (SOD-1, CAT, and GPX-1), detoxification genes (GST-1 and CYP306A1), the storage protein gene vitellogenin, and immune system antimicrobial peptide genes (defensin-1, abaecin, hymenoptaecin, and apidaecin), through real-time PCR. All fungicide treatments induced changes in gene expression, with bixafen showing the most prominent upregulation. Exposure to 1 µg of each of the three pesticides resulted in upregulation of genes associated with detoxification and nutrition processes, and downregulation of immune system genes. When the three pesticides were combined at a dose of 7 µg each, there was a pronounced downregulation of all genes. Food availability in the colonies affected the impact of fungicides on the expression of the studied genes in forager bees.

Generation of New β-Conglycinin-Deficient Soybean Lines by Editing the lincRNA lincCG1 Using the CRISPR/Cas9 System.

Soybean β-conglycinin is a major allergen that adversely affects the nutritional properties of soybean. Soybean deficient in β-conglycinin is associated with low allergenicity and high nutritional value. Long intergenic noncoding RNAs (lincRNAs) regulate gene expression and are considered important regulators of essential biological processes. Despite increasing knowledge of the functions of lincRNAs, relatively little is known about the effects of lincRNAs on the accumulation of soybean β-conglycinin. The current study presents the identification of a lincRNA lincCG1 that was mapped to the intergenic noncoding region of the β-conglycinin α-subunit locus. The full-length lincCG1 sequence was cloned and found to regulate the expression of soybean seed storage protein (SSP) genes via both cis- and trans-acting regulatory mechanisms. Loss-of-function lincCG1 mutations generated using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system led to the deficiency of the allergenic α'-, α-, and β-subunits of soybean β-conglycinin as well as higher content of proteins, sulfur-containing amino acids, and free arginine. The dominant null allele LincCG1, and consequently, the β-conglycinin-deficient phenotype associated with the lincCG1-gene-edited line was stably inherited by the progenies in a Mendelian fashion. The dominant null allele LincCG1 may therefore be exploited for engineering/developing novel hypoallergenic soybean varieties. Furthermore, Cas9-free and β-conglycinin-deficient homozygous mutant lines were obtained in the T1 generation. This study is the first to employ the CRISPR/Cas9 technology for editing a lincRNA gene associated with the soybean allergenic protein β-conglycinin. Moreover, this study reveals that lincCG1 plays a crucial role in regulating the expression of the β-conglycinin subunit gene cluster, besides highlighting the efficiency of employing the CRISPR/Cas9 system for modulating lincRNAs, and thereby regulating soybean seed components.

Storage protein activator controls grain protein accumulation in bread wheat in a nitrogen dependent manner

The expression of cereal grain storage protein (GSP) genes is controlled by a complex network of transcription factors (TFs). Storage protein activator (SPA) is a major TF acting in this network but its specific function in wheat (Triticum aestivum L.) remains to be determined. Here we generated an RNAi line in which expression of the three SPA homoeologs was reduced. In this line and its null segregant we analyzed GSP accumulation and expression of GSP and regulatory TF genes under two regimes of nitrogen availability. We show that down regulation of SPA decreases grain protein concentration at maturity under low but not high nitrogen supply. Under low nitrogen supply, the decrease in SPA expression also caused a reduction in the total quantity of GSP per grain and in the ratio of GSP to albumin-globulins, without significantly affecting GSP composition. The slight reduction in GSP gene expression measured in the SPA RNAi line under low nitrogen supply did not entirely account for the more significant decrease in GSP accumulation, suggesting that SPA regulates additional levels of GSP synthesis. Our results demonstrate a clear role of SPA in the regulation of grain nitrogen metabolism when nitrogen is a limiting resource.

Mutation of storage protein gene using CRISPR/Cas9 removed α′-subunit of β-conglycinin in soybean seeds

Mutation of storage protein gene using CRISPR/Cas9 removed α′-subunit of β-conglycinin in soybean seeds

Fusing the 3’UTR of seed storage protein genes leads to massive recombinant protein accumulation in seeds

The demand for recombinant proteins is rising dramatically, and effective production systems are currently being developed. The production of recombinant proteins in plants is a promising approach due to its low cost and low risk of contamination of the proteins with endotoxins or infectious agents from the culture serum. Plant seeds primarily accumulate seed storage proteins (SSPs), which are transcribed and translated from a few genes; therefore, the mechanism underlying SSP accumulation has been studied to help devise ways to increase recombinant protein production. We found that the 3’UTR of SSP genes are essential for SSP accumulation and can be used in the production of recombinant proteins in Arabidopsis. Fusion of the 3’UTR of SSP genes to the 3’ ends of DNA sequences encoding recombinant proteins enables massive accumulation of recombinant proteins with enzymatic activity in Arabidopsis seeds. This method is also applicable to the production of human Interferon Lambda-3 (IFN-lambda 3), a candidate biopharmaceutical compound against hepatitis C infection. Considering the low cost and ease of protein production in Arabidopsis, as well as the rapid growth of this plant, our method is useful for large-scale preparation of recombinant proteins for both academic research and biopharmaceutical production.

Dynamics of Rye Translocations Frequency in Russian Common Wheat <i>Triticum aestivum</i> L. Cultivars

The widespread using 1RS/1BL and 1RS/1AL translocation cultivars in wheat breeding was due to their high complex resistance provided by genes on 1RS chromosomes. Numerous data on the emergence of new virulent pathogen races that caused the loss of protective properties by genes on 1RS make it relevant to analyse the prospects for using both rye translocations in modern breeding programs. The use of various genetic sources of the 1RS rye chromosome in introgressive hybridization poses the problem of studying the diversity of 1RS linkage groups. We have studied the rye translocations dynamics in 240 genotypes of the wheat cultivars from 3 breeding centers of the Russian Federation. Multiple alleles of storage protein genes specific for 1RS were used to mark 1RS. The effectiveness of 1RS resistance genes to a number of pathogens, in particular to powdery mildew and stem rust, has been established. The grain quality dynamics of the 1RS/1BL cultivars has been traced. A statistically significant grain quality improvement of winter wheat cultivars since the 2000s has been confirmed. The spring cultivars quality does not depend on the presence of rye translocations in the genotypes. Accumulation of cultivars with TR:1RS/1BL observed in all three centers. A high allelic diversity of the secalin coding locus on 1RS chromosome was shown. However, all cultivars with rye translocations, regardless of 1RS origin, carried the same allele. Possible reasons for this uniformity are discussed. Translocation 1RS/1AL was not detected in the studied cultivars. Possible reasons are discussed.

Quality and Agronomic Trait Analyses of Pyramids Composed of Wheat Genes NGli-D2, Sec-1s and 1Dx5+1Dy10.

Due to rising living standards, it is important to improve wheat's quality traits by adjusting its storage protein genes. The introduction or locus deletion of high molecular weight subunits could provide new options for improving wheat quality and food safety. In this study, digenic and trigenic wheat lines were identified, in which the 1Dx5+1Dy10 subunit, and NGli-D2 and Sec-1s genes were successfully polymerized to determine the role of gene pyramiding in wheat quality. In addition, the effects of ω-rye alkaloids during 1BL/1RS translocation on quality were eliminated by introducing and utilizing 1Dx5+1Dy10 subunits through gene pyramiding. Additionally, the content of alcohol-soluble proteins was reduced, the Glu/Gli ratio was increased and high-quality wheat lines were obtained. The sedimentation values and mixograph parameters of the gene pyramids under different genetic backgrounds were significantly increased. Among all the pyramids, the trigenic lines in Zhengmai 7698, which was the genetic background, had the highest sedimentation value. The mixograph parameters of the midline peak time (MPT), midline peak value (MPV), midline peak width (MPW), curve tail value (CTV), curve tail width (CTW), midline value at 8 min (MTxV), midline width at 8 min (MTxW) and midline integral at 8 min (MTxI) of the gene pyramids were markedly enhanced, especially in the trigenic lines. Therefore, the pyramiding processes of the 1Dx5+1Dy10, Sec-1S and NGli-D2 genes improved dough elasticity. The overall protein composition of the modified gene pyramids was better than that of the wild type. The Glu/Gli ratios of the type I digenic line and trigenic lines containing the NGli-D2 locus were higher than that of the type II digenic line without the NGli-D2 locus. The trigenic lines with Hengguan 35 as the genetic background had the highest Glu/Gli ratio among the specimens. The unextractable polymeric protein (UPP%) and Glu/Gli ratios of the type II digenic line and trigenic lines were significantly higher than those of the wild type. The UPP% of the type II digenic line was higher than that of the trigenic lines, while the Glu/Gli ratio was slightly lower than that of the trigenic lines. In addition, the celiac disease (CD) epitopes' level of the gene pyramids significantly decreased. The strategy and information reported in this study could be very useful for improving wheat processing quality and reducing wheat CD epitopes.

Weighted Gene Correlation Network Analysis (WGCNA) of Arabidopsis Somatic Embryogenesis (SE) and Identification of Key Gene Modules to Uncover SE-Associated Hub Genes.

Somatic embryogenesis (SE), which occurs naturally in many plant species, serves as a model to elucidate cellular and molecular mechanisms of embryo patterning in plants. Decoding the regulatory landscape of SE is essential for its further application. Hence, the present study was aimed at employing Weighted Gene Correlation Network Analysis (WGCNA) to construct a gene coexpression network (GCN) for Arabidopsis SE and then identifying highly correlated gene modules to uncover the hub genes associated with SE that may serve as potential molecular targets. A total of 17,059 genes were filtered from a microarray dataset comprising four stages of SE, i.e., stage I (zygotic embryos), stage II (proliferating tissues at 7 days of induction), stage III (proliferating tissues at 14 days of induction), and stage IV (mature somatic embryos). This included 1,711 transcription factors and 445 EMBRYO DEFECTIVE genes. GCN analysis identified a total of 26 gene modules with the module size ranging from 35 to 3,418 genes using a dynamic cut tree algorithm. The module-trait analysis revealed that four, four, seven, and four modules were associated with stages I, II, III, and IV, respectively. Further, we identified a total of 260 hub genes based on the degree of intramodular connectivity. Validation of the hub genes using publicly available expression datasets demonstrated that at least 78 hub genes are potentially associated with embryogenesis; of these, many genes remain functionally uncharacterized thus far. In silico promoter analysis of these genes revealed the presence of cis-acting regulatory elements, “soybean embryo factor 4 (SEF4) binding site,” and “E-box” of the napA storage-protein gene of Brassica napus; this suggests that these genes may play important roles in plant embryo development. The present study successfully applied WGCNA to construct a GCN for SE in Arabidopsis and identified hub genes involved in the development of somatic embryos. These hub genes could be used as molecular targets to further elucidate the molecular mechanisms underlying SE in plants.

Long-read-based draft genome sequence of Indian black gram IPU-94-1 'Uttara': Insights into disease resistance and seed storage protein genes.

Black gram [Vigna mungo (L.) Hepper var. mungo] is a warm-season legume highly prized for its protein content along with significant folate and iron proportions. To expedite the genetic enhancement of black gram, a high-quality draft genome from the center of origin of the crop is indispensable. Here, we established a draft genome sequence of an Indian black gram cultivar, 'Uttara' (IPU 94-1), known for its high resistance to mungbean yellow mosaic virus. Pacific Biosciences of California, Inc. (PacBio) single-molecule real-time (SMRT) and Illumina sequencing assembled a draft reference-guided assembly with a cumulative size of ∼454.4 Mb, of which, 444.4 Mb was anchored on 11 pseudomolecules corresponding to 11 chromosomes. Uttara assembly denotes features of a high-quality draft genome illustrated through high N50 value (42.88 Mb), gene completeness (benchmarking universal single-copy ortholog [BUSCO] score 94.17%) and low levels of ambiguous nucleotides (N) percent (0.0005%). Gene discovery using transcript evidence predicted 28,881 protein-coding genes, from which, ∼95% were functionally annotated. A global survey of genes associated with disease resistance revealed 119 nucleotide binding site-leucine rich repeat (NBS-LRR) proteins, while 23 genes encoding seed storage proteins (SSPs) were discovered in black gram. A large set of microsatellite loci were discovered for marker development in the crop. Our draft genome of an Indian black gram provides the foundational genomic resources for the improvement of important agronomic traits and ultimately will help in accelerating black gram breeding programs.

Genetic Background Negates Improvements in Rice Flour Characteristics and Food Processing Properties Caused by a Mutant Allele of the PDIL1-1 Seed Storage Protein Gene

Phenotypic differences among breeding lines that introduce the same superior gene allele can be a barrier to effective development of cultivars with desirable traits in some crop species. For example, a deficient mutation of the Protein Disulfide Isomerase Like 1–1 (PDIL1-1) gene can cause accumulation of glutelin seed storage protein precursors in rice endosperm, and improves rice flour characteristics and food processing properties. However, the gene must be expressed to be useful. A deficient mutant allele of PDIL1-1 was introduced into two rice cultivars with different genetic backgrounds (Koshihikari and Oonari). The grain components, agronomic traits, and rice flour and food processing properties of the resulting lines were evaluated. The two breeding lines had similar seed storage protein accumulation, amylose content, and low-molecular-weight metabolites. However, only the Koshihikari breeding line had high flour quality and was highly suitable for rice bread, noodles, and sponge cake, evidence of the formation of high-molecular-weight protein complexes in the endosperm. Transcriptome analysis revealed that mRNA levels of fourteen PDI, Ero1, and BiP genes were increased in the Koshihikari breeding line, whereas this change was not observed in the Oonari breeding line. We elucidated part of the molecular basis of the phenotypic differences between two breeding lines possessing the same mutant allele in different genetic backgrounds. The results suggest that certain genetic backgrounds can negate the beneficial effect of the PDIL1-1 mutant allele. Better understanding of the molecular basis for such interactions may accelerate future breeding of novel rice cultivars to meet the strong demand for gluten-free foods.

Reducing Nitrogen Rate and Increasing Plant Density Accomplished High Yields with Satisfied Grain Quality of Soft Wheat via Modifying the Free Amino Acid Supply and Storage Protein Gene Expression.

In a 2 yr field experiment, we investigated the combined effects of reduced nitrogen (N) rate and increased plant density on the trade-off between the grain protein content (GPC) and the grain yield (GY) in soft wheat cultivars. Reducing N application significantly decreased both GPC and GY; however, to some extent, increasing the top-dressed N ratio and plant density compensated for the GY loss. Optimizing the combination of these three factors (150 kg N ha-1 with 50% top-dressed N and 360 × 104 plants ha-1) achieved both the required lower GPC for soft wheat and relatively higher GY compared with the conventional cultivation strategy. In addition, this optimized combination downregulated 11 high-molecular-weight glutenin subunits, 8 low-molecular-weight glutenin subunits, 5 α/β-gliadins, and 2 γ-gliadins in mature grains as identified by data-independent acquisition mass spectrometry. Further analysis indicated that the relatively lower free amino acid content and downregulated expressions of the seed storage protein (SSP) synthesis-related genes in filling grains contributed to the reduction of SSP and GPC. Furthermore, the dilution effect induced by a relatively higher accumulation of starch than proteins also partially explained the reduced GPC. Unlike proteins, grain starch accumulation and content depended more on the soluble sugar availability, rather than on the starch synthesis capacity. These findings provide novel insights on simultaneous improvement in the grain quality and yield of soft wheat through synchronized manipulations of N fertilization and plant density.

Characterization of the bark storage protein gene (JcBSP) family in the perennial woody plant Jatropha curcas and the function of JcBSP1 in Arabidopsis thaliana.

BackgroundBark storage protein (BSP) plays an important role in seasonal nitrogen cycling in perennial deciduous trees. However, there is no report on the function of BSP in the perennial woody oil plant Jatropha curcas.MethodsIn this study, we identified six members of JcBSP gene family in J. curcas genome. The patterns, seasonal changes, and responses to nitrogen treatment in gene expression of JcBSPs were detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Overexpression of JcBSP1 in transgenic Arabidopsis thaliana was driven by a constitutive cauliflower mosaic virus (CaMV) 35S RNA promoter.ResultsJcBSP members were found to be expressed in various tissues, except seeds. The seasonal changes in the total protein concentration and JcBSP1 expression in the stems of J. curcas were positively correlated, as both increased in autumn and winter and decreased in spring and summer. In addition, the JcBSP1 expression in J. curcas seedlings treated with different concentrations of an NH4NO3 solution was positively correlated with the NH4NO3 concentration and application duration. Furthermore, JcBSP1 overexpression in Arabidopsis resulted in a phenotype of enlarged rosette leaves, flowers, and seeds, and significantly increased the seed weight and yield in transgenic plants.

Comparison of the Agronomic, Cytological, Grain Protein Characteristics, as Well as Transcriptomic Profile of Two Wheat Lines Derived From Wild Emmer

Two advanced wheat lines BAd7-209 and BAd23-1 without the functional gene GPC-B1 were obtained from a cross between common wheat cultivar Chuannong 16 (CN16) and wild emmer wheat accession D97 (D97). BAd7-209 showed superior quality parameters than those of BAd23-1 and CN16. We found that the components of glutenins and gliadins in BAd7-209 and BAd23-1 were similar, whereas BAd7-209 had higher amount of glutenins and gliadins than those of BAd23-1. RNA sequencing analysis on developing grains of BAd7-209 and BAd23-1 as well as their parents revealed 382 differentially expressed genes (DEGs) between the high–grain protein content (GPC) (D97 + BAd7-209) and the low-GPC (CN16 + BAd23-1) groups. DEGs were mainly associated with transcriptional regulation of the storage protein genes, protein processing in endoplasmic reticulum, and protein export pathways. The upregulated gluten genes and transcription factors (e.g., NAC, MYB, and bZIP) may contribute to the high GPC in BAd7-209. Our results provide insights into the potential regulation pathways underlying wheat grain protein accumulation and contribute to make use of wild emmer for wheat quality improvement.

Transcriptional Changes in the Developing Rice Seeds Under Salt Stress Suggest Targets for Manipulating Seed Quality.

Global sea-level rise, the effect of climate change, poses a serious threat to rice production owing to saltwater intrusion and the accompanying increase in salt concentration. The reclaimed lands, comprising 22.1% of rice production in Korea, now face the crisis of global sea-level rise and a continuous increase in salt concentration. Here, we investigated the relationship between the decrease in seed quality and the transcriptional changes that occur in the developing rice seeds under salt stress. Compared to cultivation on normal land, the japonica rice cultivar, Samgwang, grown on reclaimed land showed a greatly increased accumulation of minerals, including sodium, magnesium, potassium, and sulfur, in seeds and a reduced yield, delayed heading, decreased thousand grain weight, and decreased palatability and amylose content. Samgwang showed phenotypical sensitivity to salt stress in the developing seeds. Using RNA-seq technology, we therefore carried out a comparative transcriptome analysis of the developing seeds grown on reclaimed and normal lands. In the biological process category, gene ontology enrichment analysis revealed that the upregulated genes were closely associated with the metabolism of biomolecules, including amino acids, carboxylic acid, lignin, trehalose, polysaccharide, and chitin, and to stress responses. MapMan analysis revealed the involvement of upregulated genes in the biosynthetic pathways of abscisic acid and melatonin and the relationship of trehalose, raffinose, and maltose with osmotic stress. Interestingly, many seed storage protein genes encoding glutelins and prolamins were upregulated in the developing seeds under salt stress, indicating the negative effect of the increase of storage proteins on palatability. Transcription factors upregulated in the developing seeds under salt stress included, in particular, bHLH, MYB, zinc finger, and heat shock factor, which could act as potential targets for the manipulation of seed quality under salt stress. Our study aims to develop a useful reference for elucidating the relationship between seed response mechanisms and decreased seed quality under salt stress, providing potential strategies for the improvement of seed quality under salt stress.

Genome-wide identification of seed storage protein gene regulators in wheat through coexpression analysis.

Only a few transcriptional regulators of seed storage protein (SSP) genes have been identified in common wheat (Triticum aestivum L.). Coexpression analysis could be an efficient approach to characterize novel transcriptional regulators at the genome-scale considering the correlated expression between transcriptional regulators and target genes. As the A genome donor of common wheat, Triticum urartu is more suitable for coexpression analysis than common wheat considering the diploid genome and single gene copy. In this work, the transcriptome dynamics in endosperm of T. urartu throughout grain filling were revealed by RNA-Seq analysis. In the coexpression analysis, a total of 71 transcription factors (TFs) from 23 families were found to be coexpressed with SSP genes. Among these TFs, TuNAC77 enhanced the transcription of SSP genes by binding to cis-elements distributed in promoters. The homolog of TuNAC77 in common wheat, TaNAC77, shared an identical function, and the total SSPs were reduced by about 24% in common wheat when TaNAC77 was knocked down. This is the first genome-wide identification of transcriptional regulators of SSP genes in wheat, and the newly characterized transcriptional regulators will undoubtedly expand our knowledge of the transcriptional regulation of SSP synthesis.

Detection of silkworm (Bombyx mori) and Lepidoptera DNA in feeding stuff by real-time PCR

Two real-time PCR methods were developed for the specific detection of silkworm (Bombyx mori) as an illegal ingredient in feed. The methods, a single- and a multi-copy approach, amplify small sequences of 91 and 77 basepairs (bp) size, enabling silkworm detection in severely processed products. High specificity was demonstrated for the single-copy method, with late signals (>Ct = 36) obtained only for some crustaceans. The probability of detection of the single-copy method, relying on a unique silkworm storage protein gene, was calculated to be 11 copies, respectively 10 pg native B. mori DNA. The multi-copy method, based on the classical cytochrome oxidase (COI) barcode region, revealed sensitivities as low as 0.1 pg B. mori DNA. In addition, a previously published sequencing primer pair for the order Lepidoptera (butterflies and moths) proved to be suited for rapid screening purposes. Although the multi-copy and Lepidoptera methods are less specific, unwanted cross-reactions can be screened out with an appropriate cut-off threshold. In conclusion, appropriate real-time methods are now available for the surveillance of an important by-product from the silk industry which is playing a significant role in the world markets.

Effects of storage temperature on carotenoids and amino acids in fresh Goji berry and the related regulation mechanism

<p indent=0mm>Fresh Goji berry has become more and more popular in recent years with consumers’ increasing demand for nutritional, healthy, and natural food. However, freshly harvested Goji berry is highly perishable, deteriorating and quickly losing its commercial value in <sc>1–2 d</sc> at room temperature due to its unique structural and biological characteristics (tender, hollow with thin pericarp and high water content). In order to optimize the storage conditions for fresh Goji berry and better retain its nutritional components, Goji berry was stored at 4 and −4°C for up to <sc>21 d.</sc> The changes in fruit appearance, the evolution of the contents of zeaxanthin, β-carotene, and zeaxanthin dipalmitate, and the expressions of genes related to the synthesis, storage, and degradation of carotenoids and those related to the metabolism of amino acids were investigated in the present study. The results showed that the contents of zeaxanthin, β-carotene, and zeaxanthin dipalmitate were relatively higher in Goji berry stored at 4°C for <sc>7 d</sc> or at –4°C for <sc>21 d.</sc> As the most characteristic carotenoid in Goji berry, zeaxanthin dipalmitate was 42.14% higher at –4°C than at 4°C at the end of storage. The carotenoid synthesis genes were expressed at higher levels, while the carotenoid storage protein genes, <italic>LbHSP21</italic> and <italic>LbOR2</italic>, were expressed at significantly lower levels at 4°C. At the same time, the carotenoid cleavage enzyme genes,<italic> LbNCED6</italic>, <italic>LbCCD1</italic>, and <italic>LbCCD4</italic>, were expressed at significantly higher levels in Goji berry stored at 4°C. In particular, the expression level of <italic>LbCCD4</italic> on day 7 was 70 and 30 times higher than that on day 0 at 4 and –4°C, respectively, which may explain the lower carotenoids contents at 4°C by the end of storage. The expressions of proline synthesis genes, <italic>LbP5CS</italic> and <italic>LbOAT</italic>, increased, while that of proline degradation gene <italic>LbPRODH</italic> declined during storage. The expression level of <italic>LbOAT</italic> was significantly higher, while that of <italic>LbProDH</italic> was significantly lower at 4 than at –4°C (<italic>P</italic>< 0.01), which might facilitate the accumulation of proline in Goji berry stored at 4°C. At the same time, the continuously higher expression of aspartate kinase gene <italic>LbAK</italic> was conducive to the synthesis and accumulation of asparagine. The expression of <italic>LbC4H1</italic> increased gradually during storage, which was consistent with the changing trend of phenylalanine content. In conclusion, the contents of carotenoids and amino acids in Goji berry changed dynamically during postharvest storage, which were not only regulated by the internal gene expression but also affected by the external environment such as temperature. The results of the present study showed that Goji berry stored at –4°C had higher contents of carotenoids, a lower content of proline, and therefore, a better nutritional quality with postponed senescence and deterioration. The significantly higher expression of <italic>LbCDD4</italic> and lower expressions of carotenoid storage protein genes may be the main reason for the lower carotenoids contents at 4°C compared with at –4°C. The presented study preliminarily elucidated the possible molecular mechanism of why –4°C storage is more conducive to the accumulation of carotenoids and the delay of senescence and deterioration in Goji berry. This study provides new insights for the preservation and circulation of fresh Goji berry in the supply chain.

Expression of a Seed Storage Protein AtSSA3 of &lt;i&gt;Arabidopsis thaliana&lt;/i&gt; in &lt;i&gt;Escherichia coli&lt;/i&gt;

Mature seeds are rich in storage proteins, which not only affect the quality of seeds, but also the energy source of seed germination and seedling growth. In this study, a gene encoding a seed storage protein SSA3 from Arabidopsis thaliana was isolated. The AtSSA3 gene was expressed in Escherichia coli BL21 (DE3). The fusion protein pET-32a(+)-AtSSA3 was induced under the optimized conditions of 1mM IPTG and 25°C for 4h. Western blot analysis the optimal conditions of protein expression. This study provides a theoretical and practical basis for the further study of seed storage protein genes in the future.

Mapping genomic regions controlling agronomic traits in spring wheat under conventional and organic managements

AbstractIdentification of quantitative trait loci (QTL) associated with important traits is one of the first steps towards deploying marker‐assisted selection, but the lack of stability and consistency in identifying QTL across environments and populations remains a limitation. We conducted this study to identify QTL associated with agronomic traits in wheat (Triticum aestivum L.) across management‐specific environments. A total of 204 wheat lines derived from the ‘Peace’ × ‘Carberry’ cross were evaluated in 2016 and 2017 under conventional and organic managements in Edmonton, Canada, and genotyped with diversity arrays technology markers. Using the least‐squares means for each management system, 53 QTL were identified for nine agronomic traits, 14 of which were consistently identified in both managements. The largest QTL we identified was associated with plant height (QPht.dms‐4B), which might be the plant height‐reducing gene Rht‐B1b from ‘Carberry’. It explained 54 and 49% of the phenotypic variation in conventional and organic management, respectively. The second largest QTL was associated with gluten strength (QSds.dms‐1A) in both managements. We identified consistent QTL across both organic and conventional managements, even though they were generally minor‐effect QTL. Twelve organic management‐specific QTL were found for grain yield, days to heading and maturity, plant height, test weight, and thousand‐kernel weight, but most explained relatively low amount of phenotypic variation. The QTL identified across managements, especially the gluten strength QTL (QSds.dms‐1A), may serve as useful markers in selection. The QSds.dms‐1A region needs to be investigated further to confirm whether it is the Gli‐1 storage protein gene.

High temperature inhibits the accumulation of storage materials by inducing alternative splicing of OsbZIP58 during filling stage in rice

High temperature (HT) has an adverse effect on rice grain filling by inhibiting the accumulation of storage materials. However, the regulatory mechanism of this inhibition remains unknown. Here, we report that Opaque2 like transcription factor OsbZIP58 is a key factor regulating storage material accumulation under HT. The OsbZIP58 gene promotes expression of many seed storage protein genes and starch synthesis genes while inhibits expression of some starch hydrolyzing α-amylase genes under HT. The loss of OsbZIP58 function leads to floury and shrunken endosperms and dramatically reduced storage materials in the seeds under HT. HT is found to affect alternative splicing of OsbZIP58, promoting the formation of the truncated OsbZIP58β protein form over the full-length OsbZIP58α protein form. The OsbZIP58β form has a lower transcriptional activity than the OsbZIP58α form, especially under HT condition. Interestingly, rice varieties with less heat sensitivity have reduced alternative splicing of OsbZIP58. Therefore, OsbZIP58 is a crucial gene in regulating storage material accumulation under HT and lower alternative splicing of OsbZIP58 may contribute to heat tolerance during grain filling.