CsAGA1 and CsAGA2 Mediate RFO Hydrolysis in Partially Distinct Manner in Cucumber Fruits

A Raffinose family oligosaccharides (RFOs) is one of the major translocated sugars in the vascular bundle of cucumber, but little RFOs can be detected in fruits. Alpha-galactosidases (α-Gals) catalyze the first catabolism step of RFOs. Six α-Gal genes exist in a cucumber genome, but their spatial functions in fruits remain unclear. Here, we found that RFOs were highly accumulated in vascular tissues. In phloem sap, the stachyose and raffinose content was gradually decreased, whereas the content of sucrose, glucose and fructose was increased from pedicel to fruit top. Three alkaline forms instead of acid forms of α-Gals were preferentially expressed in fruit vascular tissues and alkaline forms have stronger RFO-hydrolysing activity than acid forms. By inducible gene silencing of three alkaline forms of α-Gals, stachyose was highly accumulated in RNAi-CsAGA2 plants, while raffinose and stachyose were highly accumulated in RNAi-CsAGA1 plants. The content of sucrose, glucose and fructose was decreased in both RNAi-CsAGA1 and RNAi-CsAGA2 plants after β-estradiol treatment. In addition, the fresh- and dry-weight of fruits were significantly decreased in RNAi-CsAGA1 and RNAi-CsAGA2 plants. In cucurbitaceous plants, the non-sweet motif within the promoter of ClAGA2 is widely distributed in the promoter of its homologous genes. Taken together, we found RFOs hydrolysis occurred in the vascular tissues of fruits. CsAGA1 and CsAGA2 played key but partly distinct roles in the hydrolysis of RFOs.

Comparative Transcriptome Analysis of Leaves of Sour Jujube Seedlings under Salt Stress

Sour jujube ( Ziziphus jujuba Mill. var. spinosa ) is a native tree species in China. It is often used as the rootstock of jujube, its fruit can also be used as medicine. So it has high scientific and ecological value. Sour jujube is mostly planted in Northwest China and has extremely high salt tolerance. But the molecular mechanism of its salt tolerance is rarely studied. This study was carried out in the laboratory of Environment and Resources College of Dalian Nationalities University, two treatments were performed on the leaves of sour jujuba seedlings, the experimental group (H3): 300 mM Nacl for 3 h, and the control group (CK): sterile water for 3 h. A total of 47.02GB of valid data and 32730 annotated genes were obtained. Based on the analysis of gene expression in the comparison group, a total of 2295 genes with significant differential expression were obtained, of which 807 were up-regulated and 1488 were down-regulated. According to gene function annotation and enrichment analysis, we finally screened 148 genes. These genes were mainly involved in signal transduction of plant hormones (38), homeostasis of cell walls (27), secondary metabolism of organic matter (32), redox reactions (20) in leaves of sour jujuba seedlings under salt stress. It was also accompanied by some stress-related transcription factors (31). In addition, under salt stress, raffinose family oligosaccharides (RFO) metabolism of sour jujube seedlings was accelerated. Studying the molecular response of sour jujuba seedlings under salt stress provides a scientific basis for its cultivation in saline areas and is beneficial to further improve the salt tolerance of grafted jujube trees.

Identification and Expression Analysis of the Genes Involved in the Raffinose Family Oligosaccharides Pathway of Phaseolus vulgaris and Glycine max

Raffinose family oligosaccharides (RFO) play an important role in plants but are also considered to be antinutritional factors. A profound understanding of the galactinol and RFO biosynthetic gene families and the expression patterns of the individual genes is a prerequisite for the sustainable reduction of the RFO content in the seeds, without compromising normal plant development and functioning. In this paper, an overview of the annotation and genetic structure of all galactinol- and RFO biosynthesis genes is given for soybean and common bean. In common bean, three galactinol synthase genes, two raffinose synthase genes and one stachyose synthase gene were identified for the first time. To discover the expression patterns of these genes in different tissues, two expression atlases have been created through re-analysis of publicly available RNA-seq data. De novo expression analysis through an RNA-seq study during seed development of three varieties of common bean gave more insight into the expression patterns of these genes during the seed development. The results of the expression analysis suggest that different classes of galactinol- and RFO synthase genes have tissue-specific expression patterns in soybean and common bean. With the obtained knowledge, important galactinol- and RFO synthase genes that specifically play a key role in the accumulation of RFOs in the seeds are identified. These candidate genes may play a pivotal role in reducing the RFO content in the seeds of important legumes which could improve the nutritional quality of these beans and would solve the discomforts associated with their consumption.

Genome-wide association mapping of lentil (Lens culinaris Medikus) prebiotic carbohydrates toward improved human health and crop stress tolerance

Lentil, a cool-season food legume, is rich in protein and micronutrients with a range of prebiotic carbohydrates, such as raffinose-family oligosaccharides (RFOs), fructooligosaccharides (FOSs), sugar alcohols (SAs), and resistant starch (RS), which contribute to lentil's health benefits. Beneficial microorganisms ferment prebiotic carbohydrates in the colon, which impart health benefits to the consumer. In addition, these carbohydrates are vital to lentil plant health associated with carbon transport, storage, and abiotic stress tolerance. Thus, lentil prebiotic carbohydrates are a potential nutritional breeding target for increasing crop resilience to climate change with increased global nutritional security. This study phenotyped a total of 143 accessions for prebiotic carbohydrates. A genome-wide association study (GWAS) was then performed to identify associated variants and neighboring candidate genes. All carbohydrates analyzed had broad-sense heritability estimates (H2) ranging from 0.22 to 0.44, comparable to those reported in the literature. Concentration ranges corresponded to percent recommended daily allowances of 2–9% SAs, 7–31% RFOs, 51–111% RS, and 57–116% total prebiotic carbohydrates. Significant SNPs and associated genes were identified for numerous traits, including a galactosyltransferase (Lcu.2RBY.1g019390) known to aid in RFO synthesis. Further studies in multiple field locations are necessary. Yet, these findings suggest the potential for molecular-assisted breeding for prebiotic carbohydrates in lentil to support human health and crop resilience to increase global food security.

Bioprocessing of Barley and Lentil Grains to Obtain In Situ Synthesis of Exopolysaccharides and Composite Wheat Bread with Improved Texture and Health Properties

A comprehensive study into the potential of bioprocessing techniques (sprouting and sourdough fermentation) for improving the technological and nutritional properties of wheat breads produced using barley and lentil grains was undertaken. Dextran biosynthesis in situ during fermentation of native or sprouted barley flour (B or SB) alone or by mixing SB flour with native or sprouted lentil flour (SB-L or SB-SL) by Weissella paramesenteroides SLA5, Weissella confusa SLA4, Leuconostoc pseudomesenteroides DSM 20193 or Weissella confusa DSM 20194 was assessed. The acidification and the viscosity increase during 24 h of fermentation with and without 16% sucrose (on flour weight), to promote the dextran synthesis, were followed. After the selection of the fermentation parameters, the bioprocessing was carried out by using Leuconostoc pseudomesenteroides DSM 20193 (the best LAB dextran producer, up to 2.7% of flour weight) and a mixture of SB-SL (30:70% w/w) grains, enabling also the decrease in the raffinose family oligosaccharides. Then, the SB-SL sourdoughs containing dextran or control were mixed with the wheat flour (30% of the final dough) and leavened with baker’s yeast before baking. The use of dextran-containing sourdough allowed the production of bread with structural improvements, compared to the control sourdough bread. Compared to a baker’s yeast bread, it also markedly reduced the predicted glycemic index, increased the soluble (1.26% of dry matter) and total fibers (3.76% of dry matter) content, giving peculiar and appreciable sensory attributes.

Assessing Dynamic Changes of Taste-Related Primary Metabolism During Ripening of Durian Pulp Using Metabolomic and Transcriptomic Analyses

Durian is an economically important fruit of Southeast Asia. There is, however, a lack of in-depth information on the alteration of its metabolic networks during ripening. Here, we annotated 94 ripening-associated metabolites from the pulp of durian cv. Monthong fruit at unripe and ripe stages, using capillary electrophoresis- and gas chromatography- time-of-flight mass spectrometry, specifically focusing on taste-related metabolites. During ripening, sucrose content increased. Change in raffinose-family oligosaccharides are reported herein for the first time. The malate and succinate contents increased, while those of citrate, an abundant organic acid, were unchanged. Notably, most amino acids increased, including isoleucine, leucine, and valine, whereas aspartate decreased, and glutamate was unchanged. Furthermore, transcriptomic analysis was performed to analyze the dynamic changes in sugar metabolism, glycolysis, TCA cycle, and amino acid pathways to identify key candidate genes. Taken together, our results elucidate the fundamental taste-related metabolism of durian, which can be exploited to develop durian metabolic and genetic markers in the future.

Highest Defoliation Tolerance in Amaranthus cruentus Plants at Panicle Development Is Associated With Sugar Starvation Responses

Defoliation tolerance (DT) in Amaranthus cruentus is known to reach its apex at the panicle emergence (PE) phase and to decline to minimal levels at flowering (FL). In this study, defoliation-induced changes were recorded in the content of non-structural carbohydrates and raffinose family oligosaccharides (RFOs), and in the expression and/or activity of sugar starvation response-associated genes in plants defoliated at different vegetative and reproductive stages. This strategy identified sugar-starvation-related factors that explained the opposite DT observed at these key developmental stages. Peak DT at PE was associated with increased cytosolic invertase (CI) activity in all organs and with the extensive induction of various class II trehalose-phosphate synthase (TPS) genes. Contrariwise, least DT at FL coincided with a sharp depletion of starch reserves and with sucrose (Suc) accumulation, in leaves and stems, the latter of which was consistent with very low levels of CI and vacuolar invertase activities that were not further modified by defoliation. Increased Suc suggested growth-inhibiting conditions associated with altered cytosolic Suc-to-hexose ratios in plants defoliated at FL. Augmented cell wall invertase activity in leaves and roots, probably acting in a regulatory rather than hydrolytic role, was also associated with minimal DT observed at FL. The widespread contrast in gene expression patterns in panicles also matched the opposite DT observed at PE and FL. These results reinforce the concept that a localized sugar starvation response caused by C partitioning is crucial for DT in grain amaranth.