Evolution of the SPX gene family and its role in the response mechanism to low phosphorus stress in self-rooted apple stock.

BACKGROUND: Phosphorus plays a key role in plant adaptation to adversity and plays a positive role in the yield and quality formation of apples. Genes of the SPX domain-containing family are widely involved in the regulation of phosphorus signalling networks. However, the mechanisms controlling phosphorus deficiency are not completely understood in self-rooted apple stock. RESULTS: In this study, 26 members of the apple SPX gene family were identified by genome-wide analysis, and further divided into four subfamilies (SPX, SPX-MFS, SPX-EXS, and SPX-RING) based on their structural features. The chromosome distribution and gene duplications of MdSPXs were also examined. The promoter regions of MdSPXs were enriched for multiple biotic/abiotic stresses, hormone responses and typical P1BS-related elements. Analysis of the expression levels of 26 MdSPXs showed that some members were remarkably induced when subjected to low phosphate (Pi) stress, and in particular MdSPX2, MdSPX3, and MdPHO1.5 exhibited an intense response to low Pi stress. MdSPX2 and MdSPX3 showed significantly divergent expression levels in low Pi sensitive and insensitive apple species. Protein interaction networks were predicted for 26 MdSPX proteins. The interaction of MdPHR1 with MdSPX2, MdSPX3, MdSPX4, and MdSPX6 was demonstrated by yeast two-hybrid assay, suggesting that these proteins might be involved in the Pi-signaling pathway by interacting with MdPHR1. CONCLUSION: This research improved the understanding of the apple SPX gene family and contribute to future biological studies of MdSPX genes in self-rooted apple stock.

Complete chloroplast genome sequences of the ornamental plant Prunus cistena and comparative and phylogenetic analyses with its closely related species.

BACKGROUND: Prunus cistena is an excellent color leaf configuration tree for urban landscaping in the world, which has purplish red leaves, light pink flowers, plant shape and high ornamental value. Genomic resources for P. cistena are scarce, and a clear phylogenetic and evolutionary history for this species has yet to be elucidated. Here, we sequenced and analyzed the complete chloroplast genome of P. cistena and compared it with related species of the genus Prunus based on the chloroplast genome. RESULTS: The complete chloroplast genome of P. cistena is a 157,935 bp long typical tetrad structure, with an overall GC content of 36.72% and higher GC content in the in the inverted repeats (IR) regions than in the large single-copy (LSC) and small single-copy (SSC) regions. It contains 130 genes, including 85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The ycf3 and clpP genes have two introns, with the longest intron in the trnK-UUU gene in the LSC region. Moreover, the genome has a total of 253SSRs, with the mononucleotide SSRs being the most abundant. The chloroplast sequences and gene arrangements of P. cistena are highly conserved, with the overall structure and gene order similar to other Prunus species. The atpE, ccsA, petA, rps8, and matK genes have undergone significant positive selection in Prunus species. P. cistena has a close evolutionary relationship with P. jamasakura. The coding and IR regions are more conserved than the noncoding regions, and the chloroplast DNA sequences are highly conserved throughout the genus Prunus. CONCLUSIONS: The current genomic datasets provide valuable information for further species identification, evolution, and phylogenetic research of the genus Prunus.

Multichromosomal mitochondrial genome of Punica granatum: comparative evolutionary analysis and gene transformation from chloroplast genomes.

BACKGROUND: Punica granatum is a fundamentally important fruit tree that has important economic, medicinal and ornamental properties. At present, there are few reports on the mitochondrial genome of pomegranate. Hence, in this study the P. granatum mitogenome was sequenced and assembled to further understanding of organization, variation, and evolution of mitogenomes of this tree species. RESULTS: The genome structure was multi-chromosomes with seven circular contigs, measuring 382,774 bp in length with a 45.91% GC content. It contained 74 genes, including 46 protein-coding genes, 25 tRNA genes, and three rRNA genes. There were 188 pairs of dispersed repeats with lengths of 30 or greater, primarily consisting of reverse complementary repeats. The mitogenome analysis identified 114SSRs and 466 RNA editing sites. Analyses of codon usage, nucleotide diversity and gene migration from chloroplast to mitochondrial were also conducted. The collinear and comparative analysis of mitochondrial structures between P. granatum and its proximal species indicated that P. granatum 'Taishanhong' was closely related to P. granatum 'Qingpitian' and Lagerstroemia indica. Phylogenetic examination based on the mitogenome also confirmed the evolutionary relationship. CONCLUSION: The results offered crucial information on the evolutionary biology of pomegranate and highlighted ways to promote the utilization of the species' germplasm.

PgMYB1 Positively Regulates Anthocyanin Accumulation by Activating PgGSTF6 in Pomegranate.

The peel color of pomegranates is an important exterior quality that determines market value. Anthocyanins are biosynthesized in the cytosol and then transported to the vacuole for storage. However, the molecular mechanism that determines the color variation between red and white pomegranates remains unclear. In this study, we identified an R2R3-MYB protein (PgMYB1) that interacts with the PgGSTF6 promoter and regulates its transcriptional expression, thus promoting the accumulation of anthocyanins in pomegranate. The expression of PgMYB1 and PgGSTF6 was positively correlated with the anthocyanin content in red and white pomegranates. Further investigation showed that the knockdown of PgMYB1 in red pomegranate 'Taishanhong' (TSH), by the virus-induced gene-silencing system, inhibited anthocyanin accumulation. Together, our results indicate that PgMYB1 controls the transport of anthocyanin via PgGSTF6 and thus promotes anthocyanin accumulation in red pomegranates. Our results have a certain reference value for further clarifying the regulation of anthocyanin synthesis and transport in pomegranate fruits.

Comprehensive analysis of cucumber RAV family genes and functional characterization of CsRAV1 in salt and ABA tolerance in cucumber.

The RAV (related to ABI3 and VP1) transcription factors are specific and exist in plants, which contain a B3 DNA binding domain and/or an APETALA2 (AP2) DNA binding domain. RAVs have been extensively studied in plants, and more and more evidences show that RAVs are involved in various aspects of plant growth and development, stress resistance and hormone signal transduction. However, the systematic analysis of RAV family in cucumber is rarely reported. In this study, eight CsRAV genes were identified in cucumber genome and we further comprehensively analyzed their protein physicochemical properties, conserved domains, gene structure and phylogenetic relationships. The synteny analysis and gene duplications of CsRAV genes were also analysed. Cis-element analysis revealed that the CsRAVs promoter contained several elements related to plant hormones and abiotic stress. Expression analysis showed that NaCl and ABA could significantly induce CsRAV genes expression. Subcellular localization revealed that all CsRAVs were localized in the nucleus. In addition, 35S:CsRAV1 transgenic Arabidopsis and cucumber seedlings enhanced NaCl and ABA tolerance, revealing CsRAV1 may be an important regulator of abiotic stress response. In conclusion, comprehensive analysis of CsRAVs would provide certain reference for understanding the evolution and function of the CsRAV genes.

MdFLP enhances drought tolerance by regulating MdNAC019 in self-rooted apple stocks.

Self-rooted apple stocks are widely used for the production of apples worldwide. However, self-rooted apple stocks are weak due to shallow roots and poor grounding, resulting in poor drought resistance. Therefore, it is essential to understand the molecular mechanisms to develop self-rooted apple stock cultivars with drought resistance. We reported that MdFLP, an R2R3-MYB transcription factor, directly binds to the promoter of MdNAC019, activating its transcription and consequently enhancing drought tolerance in self-rooted apple stocks. In addition, MdFLP indirectly activates the transcriptional expression of abiotic stress-related genes, namely, MdERF6 and MdZAT10. The plants overexpressing MdFLP displayed stronger drought tolerance, whereas MdFLP-RNAi plants showed weak drought tolerance compared with non-transgenic "Gala" plants, indicating that MdFLP regulates drought tolerance in self-rooted apple stocks. Altogether, we believe that our findings provide novel insights into the functions of MdFLP in the regulation of drought tolerance in self-rooted apple stocks.

Transcriptomics and metabolomics analyses identified key genes associated with sugar and acid metabolism in sweet and sour pomegranate cultivars during the developmental period.

Pomegranate (Punica granatum), an important fruit tree in the world, is rich in bioactive substances and has broad prospects for development. In this study, gene expression levels and the concentrations of metabolites involved in the metabolism of soluble sugars and organic acids were investigated in sweet and sour pomegranate cultivars at the S1 (July 25) stage, S2 (August 26) stage, and S3 (September 24) stage. The results showed that glucose, fructose, citric acid, and malic acid were predominantly present in pomegranate. The expression of invertase 2 (INV2), INV1, FRK2, FRK7, PFK2, PFK7, and HK1 was closely correlated with the fructose and glucose contents during different developmental stages, whereas the expression of sucrose synthase 3 (SUS3) and INV1 was negatively correlated with the sucrose content. The expression of MDH (c28468_g3) and WRKY42 (c20711_g1) genes were closely related to the content of sucrose, malic acid, citric acid, and succinic acid during different developmental stages. Gene expression and metabolite concentrations varied between the two cultivars. The results provide valuable information for gene discovery, marker-assisted selection, and investigation of metabolism mechanisms in pomegranate fruits.

A Multipopulation Dynamic Adaptive Coevolutionary Strategy for Large-Scale Complex Optimization Problems.

In this paper, a multipopulation dynamic adaptive coevolutionary strategy is proposed for large-scale optimization problems, which can dynamically and adaptively adjust the connection between population particles according to the optimization problem characteristics. Based on analysis of the network evolution characteristics of collaborative search between particles, a dynamic adaptive evolutionary network (DAEN) model with multiple interconnection couplings is established in this algorithm. In the model, the swarm type is divided according to the judgment threshold of particle types, and the dynamic evolution of collaborative topology in the evolutionary process is adaptively completed according to the coupling connection strength between different particle types, which enhances the algorithm's global and local searching capability and optimization accuracy. Based on that, the evolution rules of the particle swarm dynamic cooperative search network were established, the search algorithm was designed, and the adaptive coevolution between particles in different optimization environments was achieved. Simulation results revealed that the proposed algorithm exhibited a high optimization accuracy and converging rate for high-dimensional and large-scale complex optimization problems.

Stability study of the compatibility of three chemotherapeutic agents in a mixture solution of different kinds of vehicles in the EPOCH chemotherapy scheme.

BACKGROUND: To explore the stability of a mixture of three drugs including vindesine, etoposide, and epirubicin, assigned to infusion in an EPOCH chemotherapy regimen and provide a basis for clinical use. METHODS: After mixing the three chemotherapy drugs with 500 mL of 0.9% sodium chloride or 5% glucose injection, respectively, they were divided into four groups of test solution. According to the Pharmacopoeia of the people's Republic of China, 2020 Edition, injection fluid should be tested for content, osmolarity, insoluble microparticles and pH, as well as for sterility, bacterial endotoxin and pyrogen, etc. since this experiment focuses on the compatibility of the mixture of the three drugs, sterility and the detection of bacterial endotoxin and pyrogen, etc. were not performed. The test solutions were placed at room temperature, the content was determined using high-performance liquid chromatography, and the pH, osmolarity, and insoluble microparticle changes of the mixed solution were determined. Both imported and domestic epirubicin was used. RESULTS: The four groups of test solution have no significant changes in pH, osmolarity, and insoluble microparticles were observed within 48 h, with the contents changing by less than 5%. Compared with the other three groups, the imported epirubicin saline group achieved better results with significant differences in insoluble microparticle detection items of ≥10 and ≥25 µM (P<0.05). CONCLUSIONS: The stability of the three drugs in 500 mL 0.9% sodium chloride and 5% glucose injection at room temperature was good. Imported epirubicin had some advantages in the number of insoluble microparticles and its pH was more suitable when normal saline was used as a vehicle. To reduce irritation to blood vessels by infusion, it is recommended to choose imported epirubicin with 0.9% sodium chloride mixed deployment.

The first complete chloroplast genome of Hovenia acerba Lindl.

Hovenia acerba Lindl. is an important medicinal plant, for which complete chloroplast genome (Accession: MN782301) was sequenced, assembled and annotated. The genome size is 161,668 bp and the overall GC content is 36.69%, with large single-copy (LSC, 89,451bp) regions, small single-copy (SSC, 18,979 bp) regions, and two inverted repeat regions (IRs, 26,619 bp each). A total of 130 genes are successfully annotated, including 85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The phylogenetic relationships showed that H. acerba is closely related to the species of Ziziphus genus.

Characterization of the complete chloroplast genome of Malus spectabilis 'Guanghui'.

Malus spectabilis 'Guanghui' is an important ornamental plant, which complete chloroplast genome (Accession: MT501657) was sequenced, assembled and annotated. The genome size is 1601,230 bp and the overall GC content is 36.50%, with large single-copy (LSC, 89,310bp) regions, small single-copy (SSC, 19,196 bp) regions, and two inverted repeat regions (IRs, 23,632bp each). A total of 129 genes are successfully annotated, including 84 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The phylogenetic relationships showed that Malus spectabilis 'Guanghui' is closely related to the species of Malus sieversii.

The first complete chloroplast genome of Quercus coccinea (Scarlet Oak) and its phylogenetic position within Fagaceae.

Quercus coccinea is a native to North America, which grows well in acidic soil and resists cold to extreme low temperature -40 °C. The chloroplast (cp) genome of Q. coccinea, sequenced based on next-generation platform (NEOSAT), is 161,298 bp in size. The cp genome encodes 133 genes, including 88 protein-coding genes (PCGs), 8 rRNA genes, and 37 tRNA genes. Phylogenetic relationship analysis based on complete cp genome sequences exhibited that both of Q. coccinea and Q. rubra were phylogenetically closer to Q. aliena var. Acutiserrata.

The complete chloroplast genome of Quercus robur 'Fastigiata'.

The Quercus robur 'Fastigiata' is an important ornamental plant, in which the complete chloroplast genome (accession no. MN562095) was identified and sequenced. The genome size is 161,172 bp, with a large single-copy (LSC, 90,505 bp) region, a small single-copy (SSC, 18,997 bp) region, and two inverted repeat regions (IRs, 25,835 bp each). A total of 134 genes are successfully annotated, including 89 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The phylogenetic relationships inferred that Q. robur 'Fastigiata' is closely related to Quercus mongolica, Quercus wutaishanica, and Quercus dentata.

Characterization and phylogenetic analysis of the complete chloroplast genome of Cercis canadensis 'Forest Pansy'.

Cercis canadensis 'Forest Pansy' is a tree species with high ornamental value, which complete chloroplast (cp) genome was sequenced, assembled, and annotated. The genome size is 158,960 bp with a total GC content of 36.17%. The cp genome is made up of a large single-copy region (88,114 bp), a small single-copy region (19,590 bp), and two inverted repeat regions (25,628 bp each). It contains 128 genes, including 84 protein-coding genes, 36 tRNA genes, and 8 rRNA genes. Eighteen genes were duplicated in IRs. The maximum-likelihood (ML) phylogenetic analysis indicated that the Leguminosae species are grouped together, and C. canadensis 'Forest Pansy' is closely related to C. canadensis. The result would provide valuable information for genetic studies on Cercis genus.

Grapevine microRNAs responsive to exogenous gibberellin.

BACKGROUND: MicroRNAs (miRNAs), involving in various biological and metabolic processes, have been discovered and analyzed in quite a number of plants species, such as Arabidopsis, rice and other plants. However, there have been few reports about grapevine miRNAs in response to gibberelline (GA3). RESULTS: Solexa technology was used to sequence small RNA libraries constructed from grapevine berries treated with GA3 and the control. A total of 122 known and 90 novel grapevine miRNAs (Vvi-miRNAs) were identified. Totally, 137 ones were found to be clearly responsive to GA3, among which 58 were down-regulated, 51 were up-regulated, 21 could only be detected in the control, and seven were only detected in the treatment. Subsequently, we found that 28 of them were differentially regulated by GA3, with 12 conserved and 16 novel Vvi-miRNAs, based on the analysis of qRT-PCR essays. There existed some consistency in expression levels of GA3-responsive Vvi-miRNAs between high throughput sequencing and qRT-PCR essays. In addition, 117 target genes for 29 novel miRNAs were predicted. CONCLUSIONS: Deep sequencing of short RNAs from grapevine berries treated with GA3 and the control identified 137 GA3-responsive miRNAs, among which 28 exhibited different expression profiles of response to GA3 in the diverse developmental stages of grapevine berries. These identified Vvi-miRNAs might be involved in the grapevine berry development and response to environmental stresses.

Population genetic diversity in Chinese pomegranate (Punica granatum L.) cultivars revealed by fluorescent-AFLP markers.

Eighty-five pomegranate (Punica granatum L.) cultivars from six geographical populations located at Shandong, Anhui, Shaanxi, Henan, Yunnan, and Xinjiang Provinces were studied for its population genetic diversity by means of fluorescent-AFLP markers. The results indicated that 135-185 polymorphic loci were amplified by eight pairs of primers at species level. An average of 158.25 polymorphic loci was amplified for each primer combination. The polymorphism percentage ranged from 62.5% to 86.11%, and the average polymorphism percentage was 73.26%. This indicated that there was plentiful genetic diversity in pomegranate cultivars. The genetic diversity at the species level was higher than that at the population level. The order of the genetic diversity was Henan population > Xinjiang population > Shaanxi population > Anhui population > Shandong population > Yunnan population. Variance analysis showed that there was significant difference between populations in genetic diversity. The genetic differentiation coefficient between populations (G(ST)) was 0.2018, which indicated that gene differentiation was mainly within the population, and between populations, it accounted for 20.18% of the total variation. Gene flow (Nm) between the populations measured was 1.9027 based on the genetic differentiation coefficient between populations, indicating that there was mild gene flow between populations. The UPGMA cluster analysis showed that most accessions from the same population were clustered together, but there was partly gene exchange. All genetic parameters indicated that there was plentiful genetic diversity in pomegranate cultivars in China, of which Henan population was significantly higher than the other populations, and it had wide application foreground in pomegranate breeding in China.