Anthocyanin metabolism in Nelumbo: translational and post-translational regulation control transcription.

BACKGROUND: Lotus (Nelumbo Adans.) is used as an herbal medicine and the flowers are a source of natural flavonoids. 'Da Sajin', which was firstly found in the plateau area, is a natural mutant in flower color with red streamers dyeing around white petals. RESULTS: The LC-MS-MS results showed that eight anthocyanin compounds, including cyanidin 3-O-glucoside, cyanidin 3-O-galactoside, malvidin 3-O-galactoside, and malvidin 3-O-glucoside, were differentially enriched in red-pigmented tissues of the petals, whereas most of these metabolites were undetected in white tissues of the petals. Transcriptome profiling indicated that the relative high expression levels of structural genes, such as NnPAL, NnF3H, and NnANS, was inconsistent with the low anthocyanin concentration in white tissues. Members of the NnMYB and NnbHLH transcription factor families were presumed to play a role in the metabolic flux in the anthocyanin and proanthocyanidin biosynthetic pathway. The expression model of translational initiation factor, ribosomal proteins and SKP1-CUL1-F-box protein complex related genes suggested an important role for translational and post-translational network in anthocyanin biosynthesis. In addition, pathway analysis indicated that light reaction or photo destruction might be an important external cause for floral color determination in lotus. CONCLUSIONS: In this study, it is supposed that the natural lotus mutant 'Da Sajin' may have originated from a red-flowered ancestor. Partial loss of anthocyanin pigments in petals may result from metabolic disorder caused by light destruction. This disorder is mainly regulated at post translation and translation level, resulting in a non-inherited phenotype. These results contribute to an improved understanding of anthocyanin metabolism in lotus, and indicate that the translational and post-translational regulatory network determines the metabolic flux of anthocyanins and proanthocyanidins under specific environmental conditions.

Insights into Adaptive Regulation of the Leaf-Petiole System: Strategies for Survival of Water Lily Plants under Salt Stress.

The water lily (Nymphaea tetragona) is an ancient angiosperm that belongs to the Nymphaeaceae family. As a rooted floating-leaf plant, water lilies are generally cultivated in fresh water, therefore, little is known about their survival strategies under salt stress. Long-term salt stress causes morphological changes, such as the rapid regeneration of floating leaves and a significant decrease in leaf number and surface area. We demonstrate that salt stress induces toxicity soon after treatment, but plants can adapt by regenerating floating leaves that are photosynthetically active. Transcriptome profiling revealed that ion binding was one of the most-enriched GO terms in leaf-petiole systems under salt stress. Sodium-transporter-related genes were downregulated, whereas K+ transporter genes were both up- and downregulated. These results suggest that restricting intracellular Na+ importing while maintaining balanced K+ homeostasis is an adaptive strategy for tolerating long-term salt stress. ICP-MS analysis identified the petioles and leaves as Na-hyperaccumulators, with a maximum content of over 80 g kg-1 DW under salt stress. Mapping of the Na-hyperaccumulation trait onto the phylogenetic relationships revealed that water lily plants might have a long evolutionary history from ancient marine plants, or may have undergone historical ecological events from salt to fresh water. Ammonium transporter genes involved in nitrogen metabolism were downregulated, whereas NO3--related transporters were upregulated in both the leaves and petioles, suggesting a selective bias toward NO3- uptake under salt stress. The morphological changes we observed may be due to the reduced expression of genes related to auxin signal transduction. In conclusion, the floating leaves and submerged petioles of the water lily use a series of adaptive strategies to survive salt stress. These include the absorption and transport of ions and nutrients from the surrounding environments, and the ability to hyperaccumulate Na+. These adaptations may serve as the physiological basis for salt tolerance in water lily plants.

Natural variations in the promoter of Awn Length Inhibitor 1 (ALI-1) are associated with awn elongation and grain length in common wheat.

Wheat awn plays a vital role in photosynthesis, grain production, and drought tolerance. However, the systematic identification or cloning of genes controlling wheat awn development is seldom reported. Here, we conducted a genome-wide association study (GWAS) with 364 wheat accessions and identified 26 loci involved in awn length development, including previously characterized B1, B2, Hd, and several rice homologs. The dominant awn suppressor B1 was fine mapped to a 125-kb physical interval, and a C2 H2 zinc finger protein Awn Length Inhibitor 1 (ALI-1) was confirmed to be the underlying gene of the B1 locus through the functional complimentary test with native awnless allele. ALI-1 expresses predominantly in the developing spike of awnless individuals, transcriptionally suppressing downstream genes. ALI-1 reduces cytokinin content and simultaneously restrains cytokinin signal transduction, leading to a stagnation of cell proliferation and reduction of cell numbers during awn development. Polymorphisms of four single nucleotide polymorphisms (SNPs) located in ALI-1 promoter region are diagnostic for the B1/b1 genotypes, and these SNPs are associated with awn length (AL), grain length (GL) and thousand-grain weight (TGW). More importantly, ali-1 was observed to increase grain length in wheat, which is a valuable attribute of awn on grain weight, aside from photosynthesis. Therefore, ALI-1 pleiotropically regulates awn and grain development, providing an alternative for grain yield improvement and addressing future climate changes.

Characterization of the Mitochondrial Genome of a Wheat AL-Type Male Sterility Line and the Candidate CMS Gene.

Heterosis utilization is very important in hybrid seed production. An AL-type cytoplasmic male sterile (CMS) line has been used in wheat-hybrid seed production, but its sterility mechanism has not been explored. In the present study, we sequenced and verified the candidate CMS gene in the AL-type sterile line (AL18A) and its maintainer line (AL18B). In the late uni-nucleate stage, the tapetum cells of AL18A showed delayed programmed cell death (PCD) and termination of microspore at the bi-nucleate stage. As compared to AL18B, the AL18A line produced 100% aborted pollens. The mitochondrial genomes of AL18A and AL18B were sequenced using the next generation sequencing such as Hiseq and PacBio. It was found that the mitochondrial genome of AL18A had 99% similarity with that of Triticum timopheevii, AL18B was identical to that of Triticum aestivum cv. Chinese Yumai. Based on transmembrane structure prediction, 12 orfs were selected as candidate CMS genes, including a previously suggested orf256. Only the lines harboring orf279 showed sterility in the transgenic Arabidopsis system, indicating that orf279 is the CMS gene in the AL-type wheat CMS lines. These results provide a theoretical basis and data support to further analyze the mechanism of AL-type cytoplasmic male sterility in wheat.

Genetic Mapping of ms1s, a Recessive Gene for Male Sterility in Common Wheat.

The utilization of heterosis is an important way to improve wheat yield, and the production of wheat hybrid seeds mainly relies on male-sterile lines. Male sterility in line 15 Fan 03 derived from a cross of 72,180 and Xiaoyan 6 is controlled by a single recessive gene. The gene was mapped to the distal region of chromosome 4BS in a genetic interval of 1.4 cM and physical distance of 6.57 Mb between SSR markers Ms4BS42 and Ms4BS199 using an F2 population with 1205 individuals. Sterile individuals had a deletion of 4.57 Mb in the region presumed to carry the Ms1 locus. The allele for sterility was therefore named ms1s. Three CAPS markers were developed and verified from the region upstream of the deleted fragment and can be used for ms1s marker-assisted selection in wheat hybrid breeding. This work will enrich the utilization of male sterility genetic resources.

A wheat dominant dwarfing line with Rht12, which reduces stem cell length and affects gibberellic acid synthesis, is a 5AL terminal deletion line.

Dwarfing and semi-dwarfing are important agronomic traits that have great potential for the improvement of wheat yields. Rht12, a dominant gibberellic acid (GA)-responsive dwarfing gene from the gamma-ray-induced wheat mutant Karcagi 522M7K, is located in the long arm of chromosome 5A, which is closely linked with the locus Xwmc410. Rht12 is likely an ideal gene for GA biosynthesis and deactivation research in common wheat. However, information on the Rht12 locus and sequence is lacking. In this study, Rht12 significantly shortened stem cell length and decreased GA biosynthetic components. Using bulked segregant RNA-Seq, wheat 660k single nucleotide polymorphism chip detection, and newly developed simple sequence repeat markers, Rht12 was mapped to a 11.21-Mb region at the terminal end of chromosome 5AL, and was found to be closely linked with the Xw5ac207SSR marker with a 10.73-Mb fragment deletion in all of the homologous dwarfing plants. Transcriptome analyses of the remaining 483-kb region showed significantly higher expression of the TraesCS5A01G543100 gene encoding the GA metabolic enzyme GA 2-ß-dioxygenase in dwarfing plants than in high stalk plants, suggesting that Rht12 reduces plant height by activating TaGA2ox-A14. Taken together, our findings will promote cloning and functional studies of Rht12 in common wheat.

TaCKX gene family, at large, is associated with thousand-grain weight and plant height in common wheat.

KEY MESSAGE: We used SMRT sequencing and explored the haplotypes of TaCKX genes, linked with thousand-grain weight and plant height, and developed the functionally validated markers, which can be used in the marker-assisted breeding program. Cytokinin oxidase/dehydrogenase (CKX) enzymes catalyze the permanent degradation of cytokinins. Identification of the TaCKX alleles associated with yield traits and the development of functional markers is the first step in using these alleles in marker-assisted breeding program. To identify the alleles, we sequenced the genome fragments, containing TaCKX genes from 48 wheat genotypes, by PacBio® sequencing. Six out of 22 TaCKX genes were found polymorphic, forming 14 distinct haplotypes. Functional markers were developed and validated for all the polymorphic TaCKX genes. Four specific haplotypes, i.e., TaCKX2A_2, TaCKX4A_2, TaCKX5A_3, and TaCKX9A_2, were found significantly associated with high thousand-grain weight (TGW) and short plant height (PH) in Chinese wheat micro-core collection (MCC) and GWAS open population (GWAS-OP), whereas TaCKX1B_2 in GWAS-OP and TaCKX11A_3 in MCC were significantly associated with high TGW and short PH. The mean values of TGW and PH for cumulative favorable haplotypes from chromosome 3A, i.e., TaCKX2A_2, TaCKX4A_2, and TaCKX5A_3, were significantly higher as compared to the cumulative unfavored haplotypes, and the change was additive in manner. Frequency distribution analysis revealed that since the 1960s, the frequency of the favorable haplotypes and TGW has gradually increased in Chinese wheat cultivars. Expression profiling in the seed tissue excised at 2, 4, 6, and 8 days after anthesis depicted that the favorable haplotypes are significantly less expressive as compared to the unfavored haplotypes. We conclude that the functional markers developed in this study can be used to select the favorable haplotypes of TaCKX genes in wheat marker-assisted breeding programs.

The Safety of Consuming Water Dropwort Used to Purify Livestock Wastewater Considering Accumulated Antibiotics and Antibiotic Resistance Genes.

Research is lacking on the health risks of antibiotics and antibiotic resistance genes (ARGs) in water dropwort grown in livestock wastewater. Our results showed that antibiotics from livestock wastewater were absorbed and bioaccumulated by water dropwort. The concentration of antibiotics was higher in the roots than in the stems and leaves. The health-risk coefficients of antibiotics in water dropwort were below the threshold (<0.1), indicating that in this case study, the consumption of water dropwort used to purify livestock wastewater was safe for humans considering accumulated antibiotics. ARGs were closely correlated between livestock wastewater and water dropwort, with the results showing that all 13 ARGs detected in the livestock wastewater were also found in the water dropwort. Tetracycline resistance genes were more abundant than the other ARGs in both the livestock wastewater and water dropwort. The estimated daily intake of ARGs in water dropwort for humans ranged from 2.06 × 106 to 7.75 × 1012 copies g−1, suggesting the potential risk of intaking ARGs in water dropwort cannot be ignored. Although the safety of consuming water dropwort used to purify livestock wastewater, considering accumulated antibiotics and ARGs, was assessed in this study, more studies should be conducted to ensure we fully understand the health risks.

Phytoremediation performance of three traditional ornamental hydrophytes and the structure of their rhizosphere microorganism populations.

The use of phytoremediation technology in urban and rural landscapes can permit both aesthetic and water purification functions to be achieved sustainably. Here, the ability of three ornamental aquatic plant species (Lythrum salicaria L., Sagittaria trifolia L., and Typha orientalis C. Presl) to remove nutrients from simulated contaminated water over 35 days and the structure of their rhizosphere microorganism populations were evaluated to examine their potential to be used for landscape phytoremediation as well as determine the mechanism of nutrient removal. L. salicaria had the highest nutrient removal ability (86.91-96.96% removal efficiency of total nitrogen and 46.04-66.70% removal efficiency of total phosphorus). The population structure of rhizosphere microorganisms was mainly affected by plant species and not the nutrient level of the water body according to principal coordinates analysis and non-metric multi-dimensional scaling. Betaproteobacteriales and Chitinophagales were highly correlated with the content of nutrients in water according to redundancy analysis. The accumulation of the two orders by L. salicaria and higher biomass might explain the stronger removal ability of L. salicaria. The findings of this study indicate that these plants could enhance urban and rural water landscape design; our results also shed new light on the mechanism of phytoremediation by rhizosphere microorganisms.

Trends and Causes of Raw Water Quality Indicators in the Five Most Famous Lakes of Jiangsu Province, China.

Due to pollutants from industrial and agricultural activities, the lakes in China are faced with ecological and environmental problems. The five most famous lakes of Jiangsu Province, Taihu Lake, Gehu Lake, Gaobaoshaobo Lake, Hongze Lake, and Luoma Lake, have long-term fixed monitoring points for water body-related indicators. Over a five-year period, the monitoring showed that Gehu Lake had the highest average total nitrogen (TN) and total phosphorus (TP) concentrations among all lakes which were close to the Grade V critical value of the China's Environmental Quality Standards for Surface Water (CEQSW). The NH3-N concentrations in all lakes were Grade IV according to the China's Water Quality Standard for Drinking Water Sources (CWQSDWS) and Grade II according to the CEQSW. In addition, although TP concentrations in Taihu Lake did not exceed Grade V in the CEQSW, TP removal was the main factor controlling eutrophication. It was also found that the petroleum concentrations in all lakes were lower than the Grade I according to the CEQSW. Despite this relatively low petroleum pollution, the concentration of petroleum was negatively correlated with the phytoplankton densities in all lakes. This indicated that phytoplankton density was very sensitive to petroleum concentration. For heavy metals, the concentrations of Pb, Cu, As, and Cd in all lakes were significantly lower than Grade I (CEQSW) from 2013 to 2017. However, the accumulated heavy metals in sediments will remain an important pollution source affecting water quality and aquatic products in the future. The comprehensive pollution index analysis showed that the five lakes were often moderately polluted, indicating that the protection of lake resources in China should not be relaxed for a long time in the future.

A New Green Model for the Bioremediation and Resource Utilization of Livestock Wastewater.

The rapid growth of the livestock and poultry industries has resulted in the production of a large amount of wastewater, and the treatment of this wastewater requires sustainable and environmentally friendly approaches such as phytoremediation. A substrate-free floating wetland planted with water dropwort (Oenanthe javanica), a common vegetable in Southeast China, was constructed to purify a lagoon with anaerobically and aerobically treated swine wastewater in Suqian, China. The average removal rates of total nitrogen, ammonium nitrogen, nitrite nitrogen, and chemical oxygen demand were 79.96%, 95.04%, 86.14%, and 59.91%, respectively, after 40 days of treatment. A total of 98.18 g∙m-2 nitrogen and 19.84 g∙m-2 phosphorus were absorbed into plants per harvest through the rapid growth of water dropwort biomass, and the nitrogen accumulation ability was similar to that observed of other plants, such as water hyacinth. In addition, the edible part of water dropwort was shown to comply with the Chinese National Food Sanitation Standards and be safe for human consumption. Its low soluble sugar content also makes it a suitable addition to the daily diet. Overall, substrate-free floating constructed wetlands planted with water dropwort could be more widely used for livestock wastewater purification and could be integrated with plant-livestock production in China because of its high removal efficiency and recycling utilization of water dropwort biomass.

Abundant Allelochemicals and the Inhibitory Mechanism of the Phenolic Acids in Water Dropwort for the Control of Microcystis aeruginosa Blooms.

In recent years, with the frequent global occurrence of harmful algal blooms, the use of plant allelopathy to control algal blooms has attracted special and wide attention. This study validates the possibility of turning water dropwort into a biological resource to inhibit the growth of harmful Microcystis aeruginosa blooms via allelopathy. The results revealed that there were 33 types of allelopathic compounds in the water dropwort culture water, of which 15 were phenolic acids. Regarding water dropwort itself, 18 phenolic acids were discovered in all the organs of water dropwort via a targeted metabolomics analysis; they were found to be mainly synthesized in the leaves and then transported to the roots and then ultimately released into culture water where they inhibited M. aeruginosa growth. Next, three types of phenolic acids synthesized in water dropwort, i.e., benzoic, salicylic, and ferulic acids, were selected to clarify their inhibitory effects on the growth of M. aeruginosa and their mechanism(s) of action. It was found that the inhibitory effect of phenolic acids on the growth of M. aeruginosa increased with the increase of the exposure concentration, although the algae cells were more sensitive to benzoic acid than to salicylic and ferulic acids. Further study indicated that the inhibitory effects of the three phenolic acids on the growth of M. aeruginosa were largely due to the simultaneous action of reducing the number of cells, damaging the integrity of the cell membrane, inhibiting chlorophyll a (Chl-a) synthesis, decreasing the values of F0 and Fv/Fm, and increasing the activity of the antioxidant enzymes (SOD, POD, and CAT) of M. aeruginosa. Thus, the results of this study indicate that both culture water including the rich allelochemicals in water dropwort and biological algae inhibitors made from water dropwort could be used to control the growth of noxious algae in the future.

Characterization and phylogenetic analysis of the complete chloroplast genome of Ilex rotunda, traditional Chinese medicine plant.

Ilex rotunda is a traditional Chinese medicine plant. In this study, we characterized the complete chloroplast (cp) genome sequence of I. rotunda to investigate its phylogenetic relationship. The cp genome of I. rotunda was 157,743 bp in length with 37.62% overall GC content, including a large single-copy (LSC) region of 87,060bp, a small single-copy (SSC) region of 18,432 bp, which were separated by a pair of inverted repeats (IRS) of 26,121 bp. The cp genome contained 133 genes, including 88 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Phylogenetic analysis based on whole cp genome sequences showed that I. rotunda is closely related to I. pubescens and I. polyneura.

The complete chloroplast genome of Ilex 'Emily Bruner', Ilex cornuta 'Burfordii' × Ilex latifolia (Aquifoliaceae).

Ilex 'Emily Bruner' is an important economic plant with ornamental and ecological functions in southeastern China. In this study, we characterized the complete chloroplast (cp) genome sequence of 'Emily Bruner' to investigate its phylogenetic relationship. The entire cp genome of 'Emily Bruner' was 157,216 bp in length with 37.68% overall GC content, including a large single-copy (LSC) region of 86,721 bp and a small single-copy (SSC) region of 18,427 bp, which were separated by a pair of inverted repeats (IRs) of 52,068 bp. The cp genome contained 135 genes, including 90 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Phylogenetic analysis based on whole cp genome sequences showed that 'Emily Bruner' is closest to I. cornuta species.