Different regions and environments have critical roles on yield, main quality and industrialization of an industrial purple-fleshed sweetpotato (Ipomoea batatas L. (Lam.)) "Xuzishu8".

Purple-fleshed sweetpotato (PFSP) (Ipomoea batatas (L.) Lam), whose flesh is purple to dark purple, is a special variety type of sweetpotato, which has the characteristics of food crop, industrial crop and medicinal crop. The storage root (SR) of PFSP is rich in anthocyanins, starch, protein, soluble sugar, mineral elements, polyphenol, dietary fiber and so on, which has balanced and comprehensive nutritional value. And in recent years, its unique nutritional elements are increasingly known for their health functions. At present, there is no article on the characteristics and quality analysis of industrial xz8 variety. To explore the influence of different environments on the processing quality of xz8, we selected nine regions (Xuzhou, Jiawang, Pizhou, Xinyi, Peixian, Sihong, Yanchen, Xiangyang and Tianshui) to measure its yield and quality changes. The data demonstrated that xz8 has a very consistent high yield performance. In Tianshui, the anthocyanins, protein and minerals contents were significantly higher and yield also above average. Moreover, the variety with the lowest starch content exhibited the best taste. On the basis of the above results, it suggested that quite practicable to promote xz8 cultivation and suitable for processing in these areas. Thus, our present findings improve our understanding of xz8 variety and provide the basis for the industrial production of PFSP with strong prospects for success.

RNA-Sequencing Analysis Revealed Genes Associated with Sweet Potato (Ipomoea batatas (L.) Lam.) Responses to Stem Rot during Different Infection Stages.

The sweet potato, which is an important tuber crop in China, is susceptible to a variety of pathogens and insect pests during cultivation and production. Stem rot is a common sweet potato disease that seriously affects tuber yield and quality. Unfortunately, there have been relatively few studies on the mechanism mediating the stem rot resistance of sweet potatoes. In this study, a transcriptome sequencing analysis was completed using Xushu 48 samples at different stages (T1, T2, and T3) of the stem rot infection. The T1 vs. T2, T1 vs. T3, and T2 vs. T3 comparisons detected 44,839, 81,436, and 61,932 differentially expressed genes (DEGs), respectively. The DEGs encoded proteins primarily involved in alanine, aspartate, and glutamate metabolism (ko00250), carbon fixation in photosynthetic organisms (ko00710), and amino sugar and nucleotide sugar metabolism (ko00520). Furthermore, some candidate genes induced by phytopathogen infections were identified, including gene-encoding receptor-like protein kinases (RLK5 and RLK7), an LRR receptor-like serine/threonine protein kinase (SERK1), and transcription factors (bHLH137, ERF9, MYB73, and NAC053). The results of this study provide genetic insights that are relevant to future explorations of sweet potato stem rot resistance, while also providing the theoretical basis for breeding sweet potato varieties that are resistant to stem rot and other diseases.

Comparative Analysis of Saccharification Characteristics of Different Type Sweetpotato Cultivars.

As an important characteristic crop in China, sweetpotato plays an important role in the intake and supplement of nutrients. The saccharification characteristics of sweetpotato determine the edible quality and processing type. Exploring the saccharification characteristics of sweetpotato is of great significance to the selection of processing materials and the formation mechanism of service quality, but there are few relevant studies. A comparison study of two high saccharification varieties (Y25 and Z13) and one low saccharification variety (X27) was conducted to analyze their storage roots physical and chemical properties. The results show that the dry matter content, starch, and amylose content of Y25 and Z13 were significantly different from those of X27. Furthermore, the total amylase activity was significantly higher than that of X27. On the other hand, the starch gelatinization temperature was significantly lower than that of X27. The starch reduction in Y25 and Z13 is four times more than that in X27, and the maltose content of Y25 and Z13 is more than two times that of X27. Finally, the scores of sensory evaluation and physiological sweetness were significantly higher than those of X27. The results provide a theoretical basis for understanding the saccharification characteristics of sweetpotato varieties and are of guiding significance for the selection of sweetpotato parents.

Comparative Analysis of the PYL Gene Family in Three Ipomoea Species and the Expression Profiling of IbPYL Genes during Abiotic Stress Response in Sweetpotato.

Abscisic acid (ABA), a critical phytohormone that regulates plant development and stress response, is sensed by the ABA receptors PYR/PYL/RCAR (PYLs). The PYL genes have been widely studied in multiple plant species, while a systematic analysis of PYL genes in the genus Ipomoea remains unperformed. Here, a total of 13, 14, and 14 PYLs were identified in Ipomoea batatas, Ipomoea trifida, and Ipomoea triloba, respectively. Fragment duplication was speculated to play prominent roles in Ipomoea PYL gene expansions. These Ipomoea PYLs were classified into three subfamilies via phylogenetic analysis, which was supported by exon-intron structures and conserved motif analyses. Additionally, the interspecies collinearity analysis further depicted a potential evolutionary relationship between them. Moreover, qRT-PCR analysis showed that multiple IbPYLs are highly and differentially responsive to abiotic stress treatments, suggesting their potential roles in sweetpotato stress responses. Taken together, these data provide valuable insights into the PYLs in the genus Ipomoea, which may be useful for their further functional analysis of their defense against environmental changes.

Identification and functional characterization of a flavonol synthase gene from sweet potato [Ipomoea batatas (L.) Lam.].

Flavonol synthase (FLS) is a key enzyme of the flavonoid biosynthetic pathway, which catalyzes the conversion of dihydroflavonols into flavonols. In this study, the FLS gene IbFLS1 was cloned and characterized from sweet potato. The resulting IbFLS1 protein showed a high similarity with other plant FLSs. The conserved amino acids (HxDxnH motifs) binding ferrous iron and residues (RxS motifs) binding 2-oxoglutarate were found in IbFLS1 at conserved positions, as in other FLSs, suggesting that IbFLS1 belongs to the 2-oxoglutarate-dependent dioxygenases (2-ODD) superfamily. qRT-PCR analysis showed an organ-specific pattern of expression of the IbFLS1 gene, which was predominantly expressed in young leaves. The recombinant IbFLS1 protein could catalyze the conversion of dihydrokaempferol and dihydroquercetin to kaempferol and quercetin, respectively. The results of subcellular localization studies indicated that IbFLS1 was found mainly in the nucleus and cytomembrane. Furthermore, silencing the IbFLS gene in sweet potato changed the color of the leaves to purple, substantially inhibiting the expression of IbFLS1 and upregulating the expression of genes involved in the downstream pathway of anthocyanin biosynthesis (i.e., DFR, ANS, and UFGT). The total anthocyanin content in the leaves of the transgenic plants was dramatically increased, whereas the total flavonol content was significantly reduced. Thus, we conclude that IbFLS1 is involved in the flavonol biosynthetic pathway and is a potential candidate gene of color modification in sweet potato.

A systematical genome-wide analysis and screening of WRKY transcription factor family engaged in abiotic stress response in sweetpotato.

BACKGROUND: WRKY transcription factors play pivotal roles in regulating plant multiple abiotic stress tolerance, however, a genome-wide systematical analysis of WRKY genes in sweetpotato is still missing. RESULTS: Herein, 84 putative IbWRKYs with WRKY element sequence variants were identified in sweetpotato reference genomes. Fragment duplications, rather than tandem duplications, were shown to play prominent roles in IbWRKY gene expansion. The collinearity analysis between IbWRKYs and the related orthologs from other plants further depicted evolutionary insights into IbWRKYs. Phylogenetic relationships displayed that IbWRKYs were divided into three main groups (I, II and III), with the support of the characteristics of exon-intron structures and conserved protein motifs. The IbWRKY genes, mainly from the group Ib, displayed remarkable and diverse expression profiles under multiple abiotic stress (NaCl, PEG6000, cold and heat) and hormone (ABA, ACC, JA and SA) treatments, which were determined by RNA-seq and qRT-PCR assays, suggesting their potential roles in mediating particular stress responses. Moreover, IbWRKY58L could interact with IbWRKY82 as revealed by yeast two-hybrid based on the protein interaction network screening. And abiotic stress-remarkably induced IbWRKY21L and IbWRKY51 were shown to be localized in the nucleus and had no transactivation activities. CONCLUSION: These results provide valuable insights into sweetpotato IbWRKYs and will lay a foundation for further exploring functions and possible regulatory mechanisms of IbWRKYs in abiotic stress tolerance.

A C2-Domain Abscisic Acid-Related Gene, IbCAR1, Positively Enhances Salt Tolerance in Sweet Potato (Ipomoea batatas (L.) Lam.).

Plant C2-domain abscisic acid-related (CAR) protein family plays an important role in plant growth, abiotic stress responses, and defense regulation. In this study, we cloned the IbCAR1 by homologous cloning method from the transcriptomic data of Xuzishu8, which is a sweet potato cultivar with dark-purple flesh. This gene was expressed in all tissues of sweet potato, with the highest expression level in leaf tissue, and it could be induced by NaCl and ABA. Subcellular localization analyses indicated that IbCAR1 was localized in the nucleus and plasma membrane. The PI staining experiment revealed the distinctive root cell membrane integrity of overexpressed transgenic lines upon salt stress. Salt stress significantly increased the contents of proline, ABA, and the activity of superoxide dismutase (SOD), whereas the content of malondialdehyde (MDA) was decreased in overexpressed lines. On the contrary, RNA interference plants showed sensitivity to salt stress. Overexpression of IbCAR1 in sweet potatoes could improve the salt tolerance of plants, while the RNAi of IbCAR1 significantly increased sensitivity to salt stress in sweet potatoes. Meanwhile, the genes involved in ABA biosynthesis, stress response, and reactive oxygen species (ROS)-scavenging system were upregulated in overexpressed lines under salt stress. Taken together, these results demonstrated that IbCAR1 plays a positive role in salt tolerance by relying on the ABA signal transduction pathway, activating the ROS-scavenging system in sweet potatoes.

Improving pre-harvest sprouting resistance in rice by editing OsABA8ox using CRISPR/Cas9.

KEY MESSAGE: Knock out OsABA8ox helps improve pre-harvest spouting resistance and do not affect rice yield. Pre-harvest sprouting(PHS) is a phenomenon that the seeds of crops germinate preharvest, which reduces the yield and quality of rice. Abscisic acid(ABA) is one of the phytohormones that promotes seed dormancy. ABA8' hydroxylase is the main enzyme that can catabolism ABA in plant. There are three genes that encode ABA8' hydroxylase in rice, named OsABA8ox1, OsABA8ox2 and OsABA8ox3. In this study, we use CRISPR/Cas9 gene editing technology to target these three genes in Ningjing6 and find that the knockout transgenic lines are all significantly strengthen in seed dormancy and have no effect on the yield. By a series of quantitative experiments, we consider that after knock out OsABA8ox, the high endogenous ABA level will influence the ABA signal which suppress the substantial and energy metabolism in the seeds, and finally led to higher dormancy.

Evaluation of Physiological Coping Strategies and Quality Substances in Purple SweetPotato under Different Salinity Levels.

Although salinity stress is one of the principal abiotic stresses affecting crop yield, a suitable concentration of NaCl has proven to be useful for increasing crop quality. This study used low salinity (34 mmol/L NaCl) and high salinity (85 mmol/L) to cultivate purple sweetpotato. Using transcriptomics and metabolomics to profile the pathway indicated that glycometabolism, secondary metabolite biosynthesis and the starch catabolic process were the significant pathways under the salinity stress. Further research showed that purple sweetpotato could regulate genes related to the regulation of the cellular Na+, K+, and other ions concentration in response to the low salinity tolerance, but loses this ability under high salinity. Meanwhile, under low salinity, the activity of antioxidant enzymes and their related gene expression are maintained at a high level. The low salinity influences the monosaccharide composition as well as the content and regulation of genes related to starch synthesis. Quality analysis showed that the low salinity could increase the starch content and influence the amylopectin biosynthesis. It suggested that low salinity promotes substance accumulation. High salinity could increase the anthocyanins biosynthesis and low salinity had a significant impact on phenolic acid and flavonol. Finally, the gene expression levels also prove the low salinity could change the composition and content level of the purple sweetpotato. This study showed that an appropriate concentration of NaCl can be used as an elicitor for application in purple sweetpotato planting.

Comparative Transcriptome Profiling Reveals the Genes Involved in Storage Root Expansion in Sweetpotato (Ipomoea batatas (L.) Lam.).

Sweetpotato (Ipomoea batatas (L.) Lam.) is recognized as one of the most important root crops in the world by the Food and Agriculture Organization of the United Nations. The yield of sweetpotato is closely correlated with the rate of storage root (SR) formation and expansion. At present, most of the studies on sweetpotato SR expansion are focused on the physiological mechanism. To explore the SR expansion mechanism of sweetpotato, we performed transcriptome sequencing of SR harvested at 60, 90, 120, and 150 days after planting (DAP) to analyze two sweetpotato lines, Xuzishu 8 and its crossing progenies named Xu 18-192, which were selected from an F1 segregation population of Xuzishu 8 and Meiguohong, in which SR expansion was delayed significantly. A total of 57,043 genes were produced using transcriptome sequencing, of which 1312 were differentially expressed genes (DEGs) in four SR growth periods of the sweetpotato lines. The combination of the KEGG and trend analysis revealed several key candidate genes involved in SR expansion. The SBEI gene involved in starch metabolism, and transcription factors ARF6, NF-YB3 and NF-YB10 were all significantly up-regulated during SR expansion. The data from this study provide insights into the complex mechanisms of SR formation and expansion in sweetpotato and identify new candidate genes for increasing the yield of sweetpotato.

Comparative Analysis of Anthocyanin Compositions and Starch Physiochemical Properties of Purple-Fleshed Sweetpotato "Xuzishu8" in Desert Regions of China.

The present study was undertaken to determine the scope of sweetpotato cultivation in arid regions of China. For this purpose, we investigated yield, anthocyanin compositions and physicochemical properties of starch in purple-fleshed sweetpotato (PFSP) "Xuzishu8" under humid (zi8-X) and arid (zi8-D) environments of China. The experiment was conducted in three replications in both environments during 2019 and 2020. The yield and anthocyanidins contents of PFSP were significantly higher in the arid conditions as compared to humid. Zi8-X and zi8-D both revealed the presence of three anthocyanidins, namely, cyanidin (Cy), peonidin (Pn), and pelargonidin (Pg). Cy and Pn accounted for 36.40 and 63.54% of the total anthocyanidins in zi8-X, while in zi8-D, they were found as 26.13 and 73.80%, respectively. The quantitative analysis of these anthocyanins was performed using HPLC-ESI-MS/MS which revealed eighteen anthocyanins such as nine Cy, eight Pn and one Pg. Out of which, eleven anthocyanins showed a significant difference under both conditions. Starch and amylopectin contents were found to be increased by 15.39 and 4.71%, respectively, while the amylose concentration was reduced by 15.54% under the arid environment. The diameter of the starch granule and the peak viscosity were significantly higher under arid as compared to humid conditions. On the basis of results of this study, it seems quite practicable to develop PFSP cultivation in desert regions.

Comparative transcriptomic and proteomic analysis reveals common molecular factors responsive to heat and drought stresses in sweetpotaoto (Ipomoea batatas).

Introduction: Crops are affected by various abiotic stresses, among which heat (HT) and drought (DR) stresses are the most common in summer. Many studies have been conducted on HT and DR, but relatively little is known about how drought and heat combination (DH) affects plants at molecular level. Methods: Here, we investigated the responses of sweetpotato to HT, DR, and DH stresses by RNA-seq and data-independent acquisition (DIA) technologies, using controlled experiments and the quantification of both gene and protein levels in paired samples. Results: Twelve cDNA libraries were created under HT, DR, and DH conditions and controls. We identified 536, 389, and 907 DEGs in response to HT, DR, and DH stresses, respectively. Of these, 147 genes were common and 447 were specifically associated with DH stress. Proteomic analysis identified 1609, 1168, and 1535 DEPs under HT, DR, and DH treatments, respectively, compared with the control, of which 656 were common and 358 were exclusive to DH stress. Further analysis revealed the DEGs/DEPs were associated with heat shock proteins, carbon metabolism, phenylalanine metabolism, starch and cellulose metabolism, and plant defense, amongst others. Correlation analysis identified 6465, 6607, and 6435 co-expressed genes and proteins under HT, DR, and DH stresses respectively. In addition, a combined analysis of the transcriptomic and proteomic data identified 59, 35, and 86 significantly co-expressed DEGs and DEPs under HT, DR, and DH stresses, respectively. Especially, top 5 up-regulated co-expressed DEGs and DEPs (At5g58770, C24B11.05, Os04g0679100, BACOVA_02659 and HSP70-5) and down-regulated co-expressed DEGs and DEPs (AN3, PMT2, TUBB5, FL and CYP98A3) were identified under DH stress. Discussion: This is the first study of differential genes and proteins in sweetpotato under DH stress, and it is hoped that the findings will assist in clarifying the molecular mechanisms involved in sweetpotato resistance to heat and drought stress.

The sweetpotato GIGANTEA gene promoter is co-regulated by phytohormones and abiotic stresses in Arabidopsis thaliana.

GIGANTEA (GI) is known to play significant roles in various molecular pathways. Nevertheless, the underlying mechanism of the transcriptional regulation of GI remains obscure in sweetpotato. In the present study, a 1518-bp promoter sequence was obtained from the Ipomoea batatas GIGANTEA (IbGI) gene, and several potential cis-elements responsive to light, phytohormones and abiotic stresses were identified by in silico analysis. In order to functionally validate the IbGI promoter, the 5' deletion analysis of the promoter was performed by cloning the full-length promoter (D0) and its four deletion fragments, D1 (1235 bp), D2 (896 bp), D3 (549 bp) and D4 (286 bp), upstream of the ß-glucuronidase (GUS) reporter gene. Then, these were stably transformed in Arabidopsis plants. All transgenic seedlings exhibited stable GUS activity in the condition of control, but with decreased activity in the condition of most treatments. Interestingly, merely D1 seedlings that contained an abscisic acid responsive cis-element (ABRE-element) had an extremely powerful GUS activity under the treatment of ABA, which implies that fragment spanning nucleotides of -1235 to -896 bp might be a crucial component for the responses of ABA. Eight different types of potential transcriptional regulators of IbGI were isolated by Y1H, including TGA2.2, SPLT1 and GADPH, suggesting the complex interaction mode of protein-DNA on the IbGI promoter. Taken together, these present results help to better understand the transcriptional regulation mechanism of the IbGI gene, and provides an insight into the IbGI promoter, which can be considered as an alternation for breeding transgenic plants.

Decreased grain size1, a C3HC4-type RING protein, influences grain size in rice (Oryza sativa L.).

KEY MESSAGE: We reported that DGS1 plays a positive role in regulating grain size in rice and was regulated by OsBZR1. Grain size is an important agronomic trait that contributes to grain yield. However, the underlying molecular mechanisms that determine final grain size are still largely unknown. We isolated a rice mutant showing reduced grain size in a 60Co-irradiated variety Nanjing 35 population. We named the mutant decreased grain size1 (dgs1). Map-based cloning and subsequent transgenic CRISPR and complementation assays indicated that a mutation had occurred in LOC_Os03g49900 and that the DGS1 allele regulated grain size. DGS1 encodes a protein with a 7-transmembrane domain and C3HC4 type RING domain. It was widely expressed, especially in young tissues. DGS1 is a membrane-located protein. OsBZR1 (BRASSINAZOLE-RESISTANT1), a core transcription activator of BR signaling, also plays a positive role in grain size. We provided preliminary evidence that OsBZR1 can bind to the DGS1 promoter to activate expression of DGS1.

High-Density Single Nucleotide Polymorphisms Genetic Map Construction and Quantitative Trait Locus Mapping of Color-Related Traits of Purple Sweet Potato [Ipomoea batatas (L.) Lam.].

Flesh color (FC), skin color (SC), and anthocyanin content (AC) are three important traits being used for commodity evaluation in purple-fleshed sweet potato. However, to date, only a few reports are available on the inheritance of these traits. In this study, we used a biparental mapping population of 274 F1 progeny generated from a cross between a dark purple-fleshed (Xuzishu8) and white-fleshed (Meiguohong) sweet potato variety for genetic analyses. Correlation analysis showed a significant positive correlation among AC, SC, and FC. Medium-to-high heritability was observed for these traits. We detected single nucleotide polymorphisms (SNPs) by specific length amplified fragment sequencing (SLAF-seq) with the average sequencing depth of 51.72 and 25.76 for parents and progeny, respectively. Then we constructed an integrated genetic map consisting of 15 linkage groups (LGS) of sweet potato spanning on 2,233.66 cm with an average map distance of 0.71 cm between adjacent markers. Based on the linkage map, ten major quantitative trait loci (QTLs) associated to FC, SC, and AC were identified on LG12 between 0 and 64.97 cm distance, such as one QTL for SC and FC, respectively, which explained 36.3 and 45.9% of phenotypic variation; eight QTLs for AC, which explained 10.5-28.5% of the variation. These major QTLs were highly consistent and co-localized on LG12. Positive correlation, high heritability, and co-localization of QTLs on the same LG group confirm the significance of this study to establish a marker-assisted breeding program for sweet potato improvement.

Copper-Catalyzed Oxidative Fragmentation of Alkynes with NFSI Provides Aryl Ketones.

A copper-catalyzed oxidative cleavage reaction of alkynes using NFSI and TBHP was described. Various terminal and internal alkyne substrates were employed to render quick access to aryl ketone products in moderate to good yields. NFSI not only functioned as N-centered radical precursors but also engaged in the aryl group migration. Mechanistic studies also suggested the important role of water in the title reactions.

Rice OsBT1 regulates seed dormancy through the glycometabolism pathway.

Seed dormancy and germination in rice (Oryza sativa L.) are complex and important agronomic traits that involve a number of physiological processes and energy. A mutant named h470 selected from a60Co-radiated indica cultivar N22 population had weakened dormancy that was insensitive to Gibberellin (GA) and Abscisic acid (ABA). The levels of GA4 and ABA were higher in h470 than in wild-type (WT) plants. The gene controlling seed dormancy in h470 was cloned by mut-map and transgenesis and confirmed to encode an ADP-glucose transporter protein. A 1 bp deletion in Os02g0202400 (OsBT1) caused the weaker seed dormancy in h470. Metabolomics analyses showed that most sugar components were higher in h470 seeds than the wild type. The mutation in h470 affected glycometabolism.

[A method based on wavelet package transform for denoising electromyographic signal].

The detection and analysis of electromyographic signal is of far reaching importance for clinical diagnosis as well as for convalescence medicine. Electromyographic signal is a kind of biological signal in the background full of noise, so it is somewhat difficult to detect and extract the electromyographic signal. In this paper is discussed how to use the method of wavelet package transform to denoise the electromyographic signal on the basis of its feature and its relationship with noise. The results of simulation and application in the Electromyographic Evoked Potential Measuring System independently developed by our laboratory illuminate that the method is effective for denoising electromyographic signal.