Low-concentration exogenous 3-indoleacetic acid improves fruit-setting rate of Marsdenia tenacissima by inhibiting the expression of embryo abortion-related genes.

Marsdenia tenacissima is a medicinal plant characterized by many flowers, few fruits, and a low fruit-setting rate. Exogenous auxins can improve the fruit-setting rate of plants; however, their impacts on M. tenacissima and regulatory mechanisms remain unclear. In this study, we conducted a field experiment to determine the fruit-setting rate, seed-setting rate, fruit size, and changes in transcriptional expression of related genes by spraying 10 and 50 mg·L-1 of 3-indoleacetic acid (IAA). The control plants were sprayed with distilled water. Our results indicated that the fruit-setting rate was 0.15 when treated with 10 mg·L-1 of IAA, which was 2.76-fold higher than that of the control. Compared with that of the control, the number of differentially expressed genes (DEGs) regulated by 10 mg·L-1 of IAA was 28.6-fold higher than that regulated by 50 mg·L-1 of IAA. These DEGs were closely related to hormone metabolism and fruit development. By transcriptome analysis, spraying 10 mg·L-1 of IAA increased the expressions of STP6, MYB17, and LAX3 and reduced those of CXE18, ILR1-like 3, and SAUR50; this possibly affected the ovule, embryo, and fruit development, thereby elevating the fruit-setting rate of M. tenacissima. Our results indicated that low IAA concentration increased the fruit-setting rate of M. tenacissima, providing theoretical and practical support for promoting the seed yield of M. tenacissima.

Attapulgite amendment favors the utilization of high cadmium-contaminated soil for Erigeron breviscapus cultivation.

A practical measure of soil pollution can effectively control the utilization of contaminated soil during the remediation process. In this study, Erigeron breviscapus was used as the experimental material. Soil polluted with high concentrations of cadmium (Cd) was used to study the effects of different doses of attapulgite (AP) (0, 10, 20, and 40 kg-1 for AP0, AP10, AP20, and AP40, respectively) on the yield and quality of E. breviscapus (as measured by scutellarin), as well as soil remediation. The results showed that the yield and scutellarin content of E. breviscapus decreased by 33.4% and 78.9%, respectively, in soil contaminated with high concentrations of Cd (AP0) compared with the control soil (without Cd added). Moreover, the yield increased by 48.0% and 10.6% in AP20 and AP40, respectively, compared with AP0, and the scutellarin content increased by a factor of 2.35-2.41 in AP10, AP20, and AP40. Compared with AP0, the soil Cd content decreased by 22.5-26.2% in AP10, AP20, and AP40 and the available Cd content and acid-extractable Cd fraction in the soil also decreased. The catalase, peroxidase, superoxide dismutase activities, chlorophyll, and Fe2+ content were increased in AP10, AP20, and AP40, leading to an increased yield and scutellarin content. Overall, AP20 had the best effect on the yield, quality of E. breviscapus, and soil remediation. This study provides a practical measure to consider for concurrent benefits of pollution remediation and utilization of Cd-contaminated soil.

Effects of polyethylene and biodegradable microplastics on photosynthesis, antioxidant defense systems, and arsenic accumulation in maize (Zea mays L.) seedlings grown in arsenic-contaminated soils.

Arsenic (As) and microplastic (MP) co-exposure is a major environmental problem in terrestrial ecosystems. Polyethylene and biodegradable plastics decompose into MP particles under microbial-mediated and weathering conditions. However, the effects of MP particles on physiological responses and As accumulation in maize have not been thoroughly explored. In this study, the effects of polyethylene microplastic particles (PEMPs) and biodegradable microplastic particles (BPMPs) on As accumulation, growth and physio-biochemical performance of maize seedlings (Zea mays L.) in As-contaminated soil were investigated. Our study showed that 10 % PE reduced As content in maize seedlings leaves (roots) by 41.19(34.53) µg kg-1, compared to the control. The 10 % BP reduced As content in maize seedlings leaves (roots) by 64.24 (57.27) µg kg-1. 10 % PE (10 % BP) reduced maize seedlings leaf area, total chlorophyll content and photosynthetic rate by 5.05 % (21.68 %), 44.98 % (57.12 %) and 65.29 % (77.89 %) and increased H2O2 content by 38.04 % (179.6 %), respectively. The antioxidant defense system of maize seedlings leaves was damaged by PEMPs and As co-exposure. Maize seedlings has adapted to stress by regulating antioxidant enzyme activity and the AsA-GSH cycle under BPMPs and As co-exposure. This study provides new insights into the effects of PEMPs and BPMPs on phytotoxicity and As accumulation in As-contaminated soils. Preliminarily data suggests that BPMPs may exhibit greater toxic effects on maize seedlings than PEMPs, which warrants further exploration.

Marsdenia tenacissima genome reveals calcium adaptation and tenacissoside biosynthesis.

Marsdenia tenacissima is a medicinal plant widely distributed in the calcium-rich karst regions of southwest China. However, the lack of a reference genome has hampered the implementation of molecular techniques in its breeding, pharmacology and domestication. We generated the chromosome-level genome assembly in Apocynaceae using combined SMRT sequencing and Hi-C. The genome length was 381.76 Mb, with 98.9% of it found on 11 chromosomes. The genome contained 222.63 Mb of repetitive sequences and 21 899 predicted gene models, with a contig N50 of 6.57 Mb. Phylogenetic analysis revealed that M. tenacissima diverged from Calotropis gigantea at least 13.43 million years ago. Comparative genomics showed that M. tenacissima underwent ancient shared whole-genome duplication. This event, together with tandem duplication, contributed to 70.71% of gene-family expansion. Both pseudogene analysis and selective pressure calculations suggested calcium-related adaptive evolution in the M. tenacissima genome. Calcium-induced differentially expressed genes (DEGs) were mainly enriched in cell-wall-related processes. Domains (e.g. Fasciclin and Amb_all) and cis-elements (e.g. MYB and MYC) frequently occurred in the coding and promoter regions of cell-wall DEGs, respectively, and the expression levels of these genes correlated significantly with those of calcium-signal-related transcription factors. Moreover, calcium addition increased tenacissoside I, G and H contents. The availability of this high-quality genome provides valuable genomic information for genetic breeding and molecular design, and lends insights into the calcium adaptation of M. tenacissima in karst areas.

Characteristics of heavy metal accumulation and risk assessment in understory Panax notoginseng planting system.

Yunnan Province is the main planting area of the precious Chinese herbal medicines (CHM) Panax notoginseng; however, it locates the geological area with high soil heavy metals in China. The frequent land replacement due to continuous cropping obstacles and excessive application of chemicals makes P. notoginseng prone to be contaminated by heavy metals under the farmland P. notoginseng (FPn) planting. To overcome farmland shortage, understory P. notoginseng (UPn) was developed as a new ecological planting model featured by no chemicals input. However, this newly developed planting system requires urgently the soil-plant heavy metal characteristics and risk assessment. This study aimed to evaluate the pollution status of eight heavy metals in the tillage layer (0-20 cm), subsoil layer (20-40 cm) and the plants of UPn in Lancang County, Yunnan Province. Pollution index (Pi) showed that the contamination degree of heavy metals in the tillage layer and subsoil layer was Cd > Pb > Ni > Cu > Zn > Cr > Hg > As and Pb > Cd > Cu > Ni > Cr > Hg > Zn > As, respectively. Potential ecological risk index (PERI) for the tillage layer and subsoil layer was slight and middle, respectively. The exceeding standard rate of Cd, As, Pb, Hg, Cu in the UPn roots was 5.33%, 5.33%, 13.33%, 26.67% and 1.33%, respectively, while only Cd and Hg in the UPn leaves exceeded the standard 10% and 14%, respectively. The enrichment abilities of Cd and Hg in the roots and leaves of UPn were the strongest, while that of Pb was the weakest. The Hazard index (HI) and target hazard quotient (THQ) of eight heavy metals in the roots and leaves of UPn were less than 1.Therefore, our results prove that Upn has no human health risk and provide a scientific basis for the safety evaluation and extension of UPn.

Cold Tolerance of ScCBL6 Is Associated with Tonoplast Transporters and Photosynthesis in Arabidopsis.

Plants that are adapted to harsh environments offer enormous opportunity to understand stress responses in ecological systems. Stipa capillacea is widely distributed in the frigid and arid region of the Tibetan Plateau, but its signal transduction system under cold stress has not been characterized. In this study, we isolated a cDNA encoding the signal transduction protein, ScCBL6, from S. capillacea, and evaluated its role in cold tolerance by ectopically expressing it in Arabidopsis. Full-length ScCBL6 encode 227 amino acids, and are clustered with CBL6 in Stipa purpurea and Oryza sativa in a phylogenetic analysis. Compared with tolerance in wild-type (WT) plants, ScCBL6-overexpressing plants (ScCBL6-OXP) were more tolerant to cold stress but not to drought stress, as confirmed by their high photosynthetic capacity (Fv/Fm) and survival rate under cold stress. We further compared their cold-responsive transcriptome profiles by RNA sequencing. In total, 3931 genes were differentially expressed by the introduction of ScCBL6. These gene products were involved in multiple processes such as the immune system, lipid catabolism, and secondary metabolism. A KEGG pathway analysis revealed that they were mainly enriched in plant hormone signal transduction and biomacromolecule metabolism. Proteins encoded by differentially expressed genes were predicted to be localized in chloroplasts, mitochondria, and vacuoles, suggesting that ScCBL6 exerts a wide range of functions. Based on its tonoplast subcellular location combined with integrated transcriptome and physiological analyses of ScCBL6-OXP, we inferred that ScCBL6 improves plant cold stress tolerance in Arabidopsis via the regulation of photosynthesis, redox status, and tonoplast metabolite transporters.

Gene fusion as an important mechanism to generate new genes in the genus Oryza.

BACKGROUND: Events of gene fusion have been reported in several organisms. However, the general role of gene fusion as part of new gene origination remains unknown. RESULTS: We conduct genome-wide interrogations of four Oryza genomes by designing and implementing novel pipelines to detect fusion genes. Based on the phylogeny of ten plant species, we detect 310 fusion genes across four Oryza species. The estimated rate of origination of fusion genes in the Oryza genus is as high as 63 fusion genes per species per million years, which is fixed at 16 fusion genes per species per million years and much higher than that in flies. By RNA sequencing analysis, we find more than 44% of the fusion genes are expressed and 90% of gene pairs show strong signals of purifying selection. Further analysis of CRISPR/Cas9 knockout lines indicates that newly formed fusion genes regulate phenotype traits including seed germination, shoot length and root length, suggesting the functional significance of these genes. CONCLUSIONS: We detect new fusion genes that may drive phenotype evolution in Oryza. This study provides novel insights into the genome evolution of Oryza.

Sulfur fertiliser enhancement of Erigeron breviscapus (Asteraceae) quality by improving plant physiological responses and reducing soil cadmium bioavailability.

Erigeron breviscapus (Vant.) Hand.-Mazz. is an important medicinal plant; however, its quality is severely diminished by cadmium (Cd) pollution. Sulfur fertilisation can improve the production and application of E. breviscapus. This study examined Cd stress alleviation in the soil-plant system and determined the plant growth response after the application of sulfur fertiliser. The soil Cd concentration used in the treatments was 100 g·kg-1, and the sulfur fertiliser application rates were 0.1, 0.2, and 0.3 g·kg-1. Using pot experiments, we explored the impacts of high, medium, and low amounts of sulfur fertiliser on Cd accumulation and the quality and activity of E. breviscapus. The results showed that the application of sulfur fertiliser promoted Cd transformation to residual Cd under oxidation conditions, reducing Cd accumulation in E. breviscapus. Throughout the growth period, the application of sulfur fertiliser increased the soluble protein content and antioxidant enzyme activity, which alleviated Cd toxicity. The net photosynthetic rate, transpiration rate, intercellular CO2 concentration, chlorophyll level, and leaf width increased significantly. The biomass content of E. breviscapus also increased. Sulfur fertiliser improves the quality of herbaceous medicinal plants by reducing Cd accumulation and increasing scutellarin, chlorogenic, isochlorogenic acid B, and isochlorogenic acid C contents. A reasonable application of sulfur fertiliser is essential for improving E. breviscapus quality. This study provides a new method to reduce the ecological risk of planting herbaceous medicinal plants in Cd-contaminated soil.

[Effects of intercropping and nitrogen application on soil microbial metabolic functional diversity in maize cropping soil.] / çŽ‰ç±³é—´ä½œå’Œæ–½æ°®å¯¹åœŸå£¤å¾®ç”Ÿç‰©ä»£è°¢åŠŸèƒ½å¤šæ ·æ€§çš„å½±å“.

Soil microorganism is an important indicator of soil health and plays a critical role in biogeochemical processes. We collected soil samples from a multi-year in-situ field experiment with two cropping modes (maize monocropping and maize-potato intercropping) subjected to four nitrogen (N) levels (N0, 0 kg·hm-2; N125, 125 kg·hm-2; N250, 250 kg·hm-2; N375, 375 kg·hm-2). By using the Biolog-ECO microplate method, soil microbial metabolic activity, diversity and utilization of six carbon groups were analyzed. The results showed that N application significantly increased the average well color development (AWCD) values, Simpson and Shannon indices, with the highest value at N250 in mono- and inter-cropped soils. Moreover, N application promoted the utilization of amino acids, amines and phenolic compounds, but decreased the utilization of polymers. Compared with monocropping at the same N level, intercropping improved the AWCD values, Simpson and Shannon indices, with a maximal improvement at N250. Intercropping and nitrogen application significantly affected the metabolic activities of the six carbon-source groups. In addition, intercropping improved the utilization of labile carbon sources, such as amino acids and carbohydrates. Results of the redundancy analysis and linear regression showed that intercropping and N application elevated AWCD values by increasing soil temperature, water content, and soil organic carbon content. Therefore, the changes in soil physicochemical properties after intercropping were the main reason for the enhancement of microbial metabolic activity under N application.

High quality genome of Erigeron breviscapus provides a reference for herbal plants in Asteraceae.

Erigeron breviscapus is an important medicinal plant in Compositae and the first species to realize the whole process from the decoding of the draft genome sequence to scutellarin biosynthesis in yeast. However, the previous low-quality genome assembly has hindered the optimization of candidate genes involved in scutellarin synthesis and the development of molecular-assisted breeding based on the genome. Here, the E. breviscapus genome was updated using PacBio RSII sequencing data and Hi-C data, and increased in size from 1.2 Gb to 1.43 Gb, with a scaffold N50 of 156.82 Mb and contig N50 of 140.95 kb, and a total of 43,514 protein-coding genes were obtained and oriented onto nine pseudo-chromosomes, thus becoming the third plant species assembled to chromosome level after sunflower and lettuce in Compositae. Fourteen genes with evidence for positive selection were identified and found to be related to leaf morphology, flowering and secondary metabolism. The number of genes in some gene families involved in flavonoid biosynthesis in E. breviscapus have been significantly expanded. In particular, additional candidate genes involved in scutellarin biosynthesis, such as flavonoid-7-O-glucuronosyltransferase genes (F7GATs) were identified using updated genome. In addition, three candidate genes encoding indole-3-pyruvate monooxygenase YUCCA2 (YUC2), serine carboxypeptidase-like 18 (SCPL18), and F-box protein (FBP), respectively, were identified to be probably related to leaf development and flowering by resequencing 99 individuals. These results provided a substantial genetic basis for improving agronomic and quality traits of E. breviscapus, and provided a platform for improving other draft genome assemblies to chromosome-level.

Transcriptomic response to cold of thermophilous medicinal plant Marsdenia tenacissima.

Extracts from Marsdenia tenacissima, involving tenacissoside H, I and G, have been used as remedies of cancer, inflammation and asthma. Low temperature serves as one of the main factors constrain the planting expansion and quality of M. tenacissima, but its functional mechanism has been known scarcely for the lack of genomic information and transcriptional profile. Here we investigated the transcriptomic responses of M. tenacissima under cold stress to gain insight into the molecular mechanism of low temperature sensitivity. Total RNAs were collected from samples obtained at 4-time points (after 0, 3, 6 and 48 h cold treatments with 4 °C, respectively), then used for library construction and sequenced on the Illumina Hiseq™ 4000 platform. Passing quality assessments, 500794 transcripts, and 206137 unigenes were de novo assembly out in Trinity v2.4.0, holding contig N50 of 2566 bp and unigene mean length of 754 bp. 44.20% of assembled unigenes were annotated to the well-known public protein database on a basis of sequence similarity. Using statistical comparison of the fragments per kilo base of transcript per million reads mapped (FPKM) values between conditions, 6082 group-specific differentially expressed genes (DEGs) were identified and considered as cold-responsive genes, which contained copious transcription factors and active secondary metabolism. Among them, 43 unigenes were constantly up-regulated expression along with cold time, which mainly implicated in the biosynthesis of secondary metabolites, carbon metabolism, RNA and DNA metabolism. Conversely, 21 unigenes involved in photosynthesis, cell wall, protein degradation, and transporters were downregulated continually with cold timescale. Experimentally, MtEF1α was chosen as the best housekeeping gene. Functional enrichments found that damaging of cold stress on M. tenacissima may be ascribed to inability of photosynthesis, ribsome processing, flavonoid biosynthesis and terpenoids degradation. Correlation analysis between cold induced transcription factors and tenacissoside biosynthesis-related genes indicated that 3ß-HSD significant positively correlated with bHLH51, and 4-MSO with NF-YB, GRAS3, Trihelix, FAR1, MYB60, MYBS1, bZIP43. Further promoter clone found MYB-binding site in the promoter of 4-MSO. In view of the reported cold tolerance of MYB60, it is recommended as a potential candidate suitable for future molecular design of exaptation cultivation with high bioactive constituents.

[Effects of intercropping on soil nitrification and nitrogen supply in potato field.] / 间作对马铃薯种植土壤硝化作用和硝态氮供应的影响.

Nitrate supply in soils is essential to meet nitrogen (N) demand of crops. However, how intercropping would affect soil nitrate supply and the underlying mechanisms remain unclear. Based on a field experiment of mono- and inter-cropped potato amended with four N application rates (N0, 0 kg·hm-2; N1, 62.5 kg·hm-2; N2, 125 kg·hm-2; N3, 187.5 kg·hm-2), we analyzed the differences in soil nitrate content and intensity, nitrification potential, and ammonia oxidation gene abundance. We further explored the mechanisms underlying the effects of intercropping on nitrate supply and N regulation. The results showed that both content and intensity of soil nitrate increased with N application rate. Under the same N level, nitrate content in intercropping soil was lower than that in monocropping soil. Nitrogen application increased soil nitrification potential, with stronger effects in monocropping than that of intercropping. The amoA gene abundance of AOB in soil was greater than that of AOA. The abundance of amoA gene for both AOA and AOB were increased firstly and then decreased with increasing N application rate when potato intercropped with maize. Under the same N application rate, AOA gene and AOB gene in all treatments (except N2) in intercropping were lower than those in the monocropping. The amoA gene abundance of soil AOB and AOA decreased and nitrification potential was weakened when potato intercropped with maize, resulting in a decrease of soil nitrate content and intensity. Therefore, intercropping led to a reduction of soil nitrate supply, which was related to the microbial process of soil N transformation. Much attention should be paid to soil N supply under the condition of potato and maize intercropping.

Comparative transcriptome and metabolome analyses provide new insights into the molecular mechanisms underlying taproot thickening in Panax notoginseng.

BACKGROUND: Taproot thickening is a complex biological process that is dependent on the coordinated expression of genes controlled by both environmental and developmental factors. Panax notoginseng is an important Chinese medicinal herb that is characterized by an enlarged taproot as the main organ of saponin accumulation. However, the molecular mechanisms of taproot enlargement are poorly understood. RESULTS: A total of 29,957 differentially expressed genes (DEGs) were identified during the thickening process in the taproots of P. notoginseng. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment revealed that DEGs associated with "plant hormone signal transduction," "starch and sucrose metabolism," and "phenylpropanoid biosynthesis" were predominantly enriched. Further analysis identified some critical genes (e.g., RNase-like major storage protein, DA1-related protein, and Starch branching enzyme I) and metabolites (e.g., sucrose, glucose, fructose, malate, and arginine) that potentially control taproot thickening. Several aspects including hormone crosstalk, transcriptional regulation, homeostatic regulation between sugar and starch, and cell wall metabolism, were identified as important for the thickening process in the taproot of P. notoginseng. CONCLUSION: The results provide a molecular regulatory network of taproot thickening in P. notoginseng and facilitate the further characterization of the genes responsible for taproot formation in root medicinal plants or crops.

Illumina-based transcriptomic analysis on recalcitrant seeds of Panax notoginseng for the dormancy release during the after-ripening process.

Panax notoginseng (Burk) F.H. Chen is an economically and medicinally important plant of the family Araliacease, with seed dormancy being a key factor limiting the extended cultivation of P. notoginseng. The seeds belong to the morphophysiological dormancy (MPD) group, and it has also been described as the recalcitrant seed. To date, the molecular mechanism of dormancy release in the recalcitrant seed of P. notoginseng is unknown. In the present study, the transcript profiles of seeds from different after-ripening stages (0, 20, 40 and 60 days) were investigated using Illumina Hiseq 2500 technology. 91 979 946 clean reads were generated, and 81 575 unigenes were annotated in at least one database. In addition, the differentially expressed genes (DEGs) were identified by the pairwise comparisons. We screened out 2483 DEGs by the three key groups of 20 days vs 0 d, 40 d vs 0 d and 60 d vs 0 d. The DEGs were analyzed by gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway annotation. Meanwhile, we obtained 78 DEGs related to seeds dormancy release at different after-ripening stages of P. notoginseng, of which 15 DEGs were associated with abscisic acid and gibberellin. 26 DEGs that encode late embryogenesis abundant protein and antioxidant enzyme were correlated with desiccation tolerance in seeds. In summary, the results obtained here showed that PECTINESTERASE-2-LIKE, GA-INSENSITIVE, ENT-KAURENE SYNTHASE, PROTEIN PHOSPHATASE 2C, GIBBERELLIN 2-BETA-DIOXYGENASE, SUPEROXIDE DISMUTASE, L-ASCORBATE PEROXIDASE, CATALASE, LATE EMBRYOGENESIS ABUNDANT PROTEIN DC3 and DEHYDRIN 9 were potentially involved in dormancy release and desiccation sensitivity of P. notoginseng seeds. The data might provide a basis for researches on MPD.

Unigene-based RNA-seq provides insights on drought stress responses in Marsdenia tenacissima.

Marsdenia tenacissima is a well-known anti-cancer medicinal plant used in traditional Chinese medicine, which often grows on the karst landform and the water conservation capacity of land is very poorly and drought occurrences frequently. We found M. tenacissima has strong drought resistance because of continuousdrought16 d, the leaves of M. tenacissima were fully curly and dying. But the leaves were fully almost recovering after re-watering 24h. The activity of SOD and POD were almost doubled under drought stress. The content of osmotic regulating substance proline and soluble sugar were three times than control group. But after re-watering, these indexes were declined rapidly. Three cDNA libraries of control, drought stress, and re-watering treatments were constructed. There were 43,129,228, 47,116,844, and 42,815,454 clean reads with Q20 values of 98.06, 98.04, and 97.88respectively.SRA accession number of raw data was PRJNA498187 on NCBI. A total of 8672, 6043, and 6537 differentially expressed genes (DEGs) were identified in control vs drought stress, control vs re-watering, and drought stress vs re-watering, respectively. In addition, 1039, 1016, and 980 transcription factors (TFs) were identified, respectively. Among them, 363, 267, and 299 TFs were identified as DEGs in drought stress, re-watering, and drought stress and re-watering, respectively. These differentially expressed TFs mainly belonged to the bHLH, bZIP, C2H2, ERF, MYB, MYB-related, and NAC families. A comparative analysis found that 1174 genes were up-regulated and 2344 were down-regulated under drought stress and this pattern was the opposite to that found after re-watering. Among the up-regulated genes, 64 genes were homologous to known functional genes that directly protect plants against drought stress. Furthermore, 44 protein kinases and 38 TFs with opposite expression patterns under drought stress and re-watering were identified, which are possibly candidate regulators for drought stress resistance in M. tenacissima. Our study is the first to characterize the M. tenacissima transcriptome in response to drought stress, and will serve as a useful resource for future studies on the functions of candidate protein kinases and TFs involved in M. tenacissima drought stress resistance.

Engineering yeast for the production of breviscapine by genomic analysis and synthetic biology approaches.

The flavonoid extract from Erigeron breviscapus, breviscapine, has increasingly been used to treat cardio- and cerebrovascular diseases in China for more than 30 years, and plant supply of E. breviscapus is becoming insufficient to satisfy the growing market demand. Here we report an alternative strategy for the supply of breviscapine by building a yeast cell factory using synthetic biology. We identify two key enzymes in the biosynthetic pathway (flavonoid-7-O-glucuronosyltransferase and flavone-6-hydroxylase) from E. breviscapus genome and engineer yeast to produce breviscapine from glucose. After metabolic engineering and optimization of fed-batch fermentation, scutellarin and apigenin-7-O-glucuronide, two major active ingredients of breviscapine, reach to 108 and 185 mg l-1, respectively. Our study not only introduces an alternative source of these valuable compounds, but also provides an example of integrating genomics and synthetic biology knowledge for metabolic engineering of natural compounds.

[A semimicroquality evaluation method on Panax notoginseng and its application in analysis of continuous cropping obstacles research samples].

Panax notoginseng is a commonly used traditional Chinese medicine with blood activating effect while has continuous cropping obstacle problem in planting process. In present study, a semimicroextraction method with water-saturated n-butanol on 0.1 g notoginseng sample was established with good repeatability (RSD<2.5%) and 9.6%-20.6% higher extraction efficiency of seven saponins than the conventional method. A total of 16 characteristic peaks were identified by LC-MS-IT-TOF, including eight 20(S)-protopanaxatriol (PPT) type saponins and eight 20(S)-protopanaxadiol (PPD) type saponins. The established method was utilized to evaluate the quality of notoginseng samples cultivated by manual intervened methods to overcome continuous cropping obstacles.As a result, HPLC fingerprint similarity, content of Fa and ratio of notoginsenoside K and notoginsenoside Fa (N-K/Fa) were found out to be as valuatable markers of the quality of samples in continuous cropping obstacle research, of which N-K/Fa could also be applied to the analysis of notoginseng samples with different growth years.Notoginseng samples with continuous cropping obstacle had HPLC fingerprint similarity lower than 0.87, in consistent with normal sample, and had significant lower content of notoginsenoside Fa and significant higher N-K/Fa (2.35-4.74) than normal group (0.45-1.33). All samples in the first group with manual intervention showed high similarity with normal group (>0.87), similar content of common peaks and N-K/Fa (0.42-2.06). The content of notoginsenoside K in the second group with manual intervention was higher than normal group. All samples except two displayed similarity higher than 0.87 and possessed content of 16 saponins close to normal group. The result showed that notoginseng samples with continuous cropping obstacle had lower quality than normal sample. And manual intervened methods could improve their quality in different levels.The method established in this study was simple, fast and accurate, and the markers may provide new guides for quality control in continuous cropping obstacle research of notoginseng.

[Effect of Different Nitrogen Rates on the Nitrification Potential and Abundance of Ammonia-oxidizer in Intercropping Maize Soils].

The objectives of this study were to explore the effects of different nitrogen rates on the nitrification potential in intercropping maize soils. The experiment included two plantation types (maize monocropping and intercropping maize and potato) and four nitrogen levels (N0: no fertilizer treatment, N1: 1/2 conventional fertilizer treatment, N2: conventional fertilizer treatment, N3: 3/2 conventional fertilizer treatment). Abundance of AOA and AOB was analyzed with real-time quantitative PCR (Q-PCR) and soil properties were also measured. Results of this study suggested that the PNF of soils, the abundance of AOA and AOB were increased with increasing N fertilization in conventional fertilizer range, and there was no significant difference between N2 and N3 treatments. The influences of intercropping on PNF, AOA and AOB were associated with N application rates and crop growth period, as well as the benefits of ammonia oxidizer and nitrification of intercropping on N1 level. Fertilization was obviously the main driving force of the PNF, correlation analysis indicated that water content was the main environmental factor affecting the nitrification potential. Nitrification potential in maize and potato intercropping soils exhibited significant positive correlations with AOA and AOB amoA gene abundance. Maize and potato intercropping could decrease the abundance of AOA and AOB, but it resulted in dominance of AOB among nitrification bacteria in these soils. In conclusion, N fertilizer and intercropping could affect both soil nitrogen internal cycling process and the abundance of AOA and AOB, which further affected soil environmental quality.

Photosynthesis, light energy partitioning, and photoprotection in the shade-demanding species Panax notoginseng under high and low level of growth irradiance.

Partitioning of light energy into several pathways and its relation to photosynthesis were examined in a shade-demanding species Panax notoginseng (Burkill) F.H.Chen ex C.Y.Wu & K.M.Feng grown along a light gradient. In fully light-induced leaves, the actual efficiency of PSII photochemistry (ΔF/Fm'), electron transport rate (ETR), non-photochemical quenching (NPQ) and photochemical quenching (qP) were lower in low-light-grown plants; this was also the case in fully dark-adapted leaves under a simulated sunfleck. In response to varied light intensity, high-light-grown plants showed greater quantum yields of light-dependent non-photochemical quenching (ΦNPQ) and PSII photochemistry (ΦPSII) and smaller quantum yields of fluorescence and constitutive thermal dissipation (Φf,d). Under the simulated sunfleck, high-light-grown plants showed greater ΦPSII and smaller Φf,d. There were positive relationships between net photosynthesis (Anet) and ΦNPQ+f,d and negative relationships between Anet and ΦPSII in fully light-induced leaves; negative correlations of Anet with ΦNPQ+f,d and positive correlations of Anet with ΦPSII were observed in fully dark-adapted leaves. In addition, more nitrogen was partitioned to light-harvesting components in low-light-grown plants, whereas leaf morphology and anatomy facilitate reducing light capture in high-light-grown plants. The pool of xanthophyll pigments and the de-epoxidation state was greater in high-light-grown plants. Antioxidant defence was elevated by increased growth irradiance. Overall, the evidences from P. notoginseng suggest that in high-light-grown shade-demanding plants irradiated by high light more electrons were consumed by non-net carboxylative processes that activate the component of NPQ, that low-light-grown plants correspondingly protect the photosynthetic apparatus against photodamage by reducing the efficiency of PSII photochemistry under high light illumination, and that during the photosynthetic induction, the ΔpH-dependent (qE) component of NPQ might dominate photoprotection, but the NPQ also depresses the enhancement of photosynthesis via competition for light energy.

[Wild resources survey of Marsdenia tenacissima in Honghe, Yunnan].

To ascertain current situation of wild Marsdenia tenacissima resources in Honghe, Yunnan province, the distribution, habitat characteristic and resources reserves of M. tenacissima were surveyed based on interviews and investigation. The results showed that M. tenacissima was found in 7 counties such as Jinping, Mengzi etc, and distributed mainly on the mountainsides from 800 m to 1 200 m. And distribution was affected by many factors, such as light, heat, topography, soil, and vegetation. M. tenacissima grew well in distribution areas. M. tenacissima had averagely a weight of 2.8 kg per plant. Resources reserve of M. tenacissima in Honghe was estimated to 1 300 tons by now but it reduced rapidly in resent years, the wild resources reserve may not meet demand of market. Resources protection and wildlife tending would be conducted to deal with increasing medication requirements.