OsmiR319-OsPCF5 modulate resistance to brown planthopper in rice through association with MYB proteins.

BACKGROUND: The brown planthopper (BPH) is a kind of piercing-sucking insect specific to rice, with the damage tops the list of pathogens and insects in recent years. microRNAs (miRNAs) are pivotal regulators of plant-environment interactions, while the mechanism underlying their function against insects is largely unknown. RESULTS: Here, we confirmed that OsmiR319, an ancient and conserved miRNA, negatively regulated resistance to BPHs, with overexpression of OsmiR319 susceptible to BPH, while suppression of OsmiR319 resistant to BPH in comparison with wild type. Meanwhile, we identified several targets of OsmiR319 that may mediate BPH resistance. Among them, OsPCF5 was the most obviously induced by BPH feeding, and over expression of OsPCF5 was resistance to BPH. In addition, various biochemical assays verified that OsPCF5 interacted with several MYB proteins, such as OsMYB22, OsMYB30, and OsMYB30C.Genetically, we revealed that both OsMYB22 and OsMYB30C positively regulated BPH resistance. Genetic interaction analyses confirmed that OsMYB22 and OsMYB30C both function in the same genetic pathway with OsmiR319b to mediate BPH resistance. CONCLUSIONS: Altogether, we revealed that OsPCF5 regulates BPH resistance via association with several MYB proteins downstream of OsmiR319, these MYB proteins might function as regulators of BPH resistance through regulating the phenylpropane synthesis.

Mutation, food-grade expression, and characterization of a lactonase for zearalenone degradation.

Zearalenone (ZEN) is a mycotoxin that causes serious threats to human health. People are exposed to ZEN contamination externally and internally through many ways, while environmental-friendly strategies for efficient elimination of ZEN are urgently needed worldwide. Previous studies revealed that the lactonase Zhd101 from Clonostachys rosea can hydrolyze ZEN to low toxicity compounds. In this work, the enzyme Zhd101 was conducted with combinational mutations to enhance its application properties. The optimal mutant (V153H-V158F), named Zhd101.1, was selected and introduced into the food-grade recombinant yeast strain Kluyveromyces lactis GG799(pKLAC1-Zhd101.1), followed by induced expression and secretion into the supernatant. The enzymatic properties of this mutant were extensively examined, revealing a 1.1-fold increase in specific activity, as well as improved thermostability and pH stability, compared to the wild-type enzyme. The ZEN degradation tests and the reaction parameters optimization were carried out in both solutions and the ZEN-contaminated corns, using the fermentation supernatants of the food-grade yeast strain. Results showed that the degradation rates for ZEN by fermentation supernatants reached 96.9% under optimal reaction conditions and 74.6% in corn samples, respectively. These new results are a useful reference to zearalenone biodegradation technologies and indicated that the mutant enzyme Zhd101.1 has potential to be used in food and feed industries. KEY POINTS: • Mutated lactonase showed 1.1-fold activity, better pH stability than the wild type. • The strain K. lactis GG799(pKLAC1-Zhd101.1) and the mutant Zhd101.1 are food-grade. • ZEN degradation rates by supernatants reached 96.9% in solution and 74.6% in corns.

Integrated metabolomics and transcriptomics study of traditional herb Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao reveals global metabolic profile and novel phytochemical ingredients.

BACKGROUND: Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao is one of the most common herbs widely used in South and East Asia, to enhance people's health and reinforce vital energy. Despite its prevalence, however, the knowledge about phytochemical compositions and metabolite biosynthesis in Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao is very limited. RESULTS: An integrated metabolomics and transcriptomics analysis using state-of-the-art UPLC-Q-Orbitrap mass spectrometer and advanced bioinformatics pipeline were conducted to study global metabolic profiles and phytochemical ingredients/biosynthesis in Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao. A total of 5435 metabolites were detected, from which 2190 were annotated, representing an order of magnitude increase over previously known. Metabolic profiling of Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao tissues found contents and synthetic enzymes for phytochemicals were significantly higher in leaf and stem in general, whereas the contents of the main bioactive ingredients were significantly enriched in root, underlying the value of root in herbal remedies. Using integrated metabolomics and transcriptomics data, we illustrated the complete pathways of phenylpropanoid biosynthesis, flavonoid biosynthesis, and isoflavonoid biosynthesis, in which some were first reported in the herb. More importantly, we discovered novel flavonoid derivatives using informatics method for neutral loss scan, in addition to inferring their likely synthesis pathways in Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao. CONCLUSIONS: The current study represents the most comprehensive metabolomics and transcriptomics analysis on traditional herb Astragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao. We demonstrated our integrated metabolomics and transcriptomics approach offers great potentials in discovering novel metabolite structure and associated synthesis pathways. This study provides novel insights into the phytochemical ingredients, metabolite biosynthesis, and complex metabolic network in herbs, highlighting the rich natural resource and nutritional value of traditional herbal plants.

Stigma and Its Influence on Patients With Temporary Ostomy: A Cross-sectional Survey.

PURPOSE: The purpose of this study was to explore stigma and its influencing factors in Chinese patients with a temporary ostomy. DESIGN: Cross-sectional, descriptive. SUBJECTS AND SETTING: Convenience sampling was used to recruit 170 patients living with a temporary ostomy for at least 2 weeks. Participants were recruited from 3 general hospitals in Guangdong province, located in southeastern China. METHODS: Respondents completed a questionnaire that included ostomy-related sociodemographic questionnaire, the Social Impact Scale, and the Stoma Self-Efficacy Scale. Multivariate linear regression was used to identify factors influencing stigma. RESULTS: Of the 170 questionnaires distributed, 159 (93.5%) respondents who sufficiently completed responses were included in the final analysis. The average score of the Social Impact Scale was 61.63 (SD = 8.39), reflecting a moderate level of stigma, and 27 (17%) experienced high levels of stigma. Multivariate regression analysis revealed that stigma was associated with family relationships (B = 6.139, P = .000), age when ostomy was created (B = -4.038, P = .000), and self-efficacy (B = -0.080, P = .002). CONCLUSIONS: The level of stigma in Chinese patients with temporary ostomies is moderate. Multivariate regression analysis identified family relationship, age, and self-efficacy as the main factors that influenced stigma in patients with temporary ostomy.

Global transcriptome analysis of Huperzia serrata and identification of critical genes involved in the biosynthesis of huperzine A.

BACKGROUND: Huperzia serrata (H. serrata) is an economically important traditional Chinese herb with the notably medicinal value. As a representative member of the Lycopodiaceae family, the H. serrata produces various types of effectively bioactive lycopodium alkaloids, especially the huperzine A (HupA) which is a promising drug for Alzheimer's disease. Despite their medicinal importance, the public genomic and transcriptomic resources are very limited and the biosynthesis of HupA is largely unknown. Previous studies on comparison of 454-ESTs from H. serrata and Phlegmariurus carinatus predicted putative genes involved in lycopodium alkaloid biosynthesis, such as lysine decarboxylase like (LDC-like) protein and some CYP450s. However, these gene annotations were not carried out with further biochemical characterizations. To understand the biosynthesis of HupA and its regulation in H. serrata, a global transcriptome analysis on H. Serrata tissues was performed. RESULTS: In this study, we used the Illumina Highseq4000 platform to generate a substantial RNA sequencing dataset of H. serrata. A total of 40.1 Gb clean data was generated from four different tissues: root, stem, leaf, and sporangia and assembled into 181,141 unigenes. The total length, average length, N50 and GC content of unigenes were 219,520,611 bp, 1,211 bp, 2,488 bp and 42.51%, respectively. Among them, 105,516 unigenes (58.25%) were annotated by seven public databases (NR, NT, Swiss-Prot, KEGG, COG, Interpro, GO), and 54 GO terms and 3,391 transcription factors (TFs) were functionally classified, respectively. KEGG pathway analysis revealed that 72,230 unigenes were classified into 21 functional pathways. Three types of candidate enzymes, LDC, CAO and PKS, responsible for the biosynthesis of precursors of HupA were all identified in the transcripts. Four hundred and fifty-seven CYP450 genes in H. serrata were also analyzed and compared with tissue-specific gene expression. Moreover, two key classes of CYP450 genes BBE and SLS, with 23 members in total, for modification of the lycopodium alkaloid scaffold in the late two stages of biosynthesis of HupA were further evaluated. CONCLUSION: This study is the first report of global transcriptome analysis on all tissues of H. serrata, and critical genes involved in the biosynthesis of precursors and scaffold modifications of HupA were discovered and predicted. The transcriptome data from this work not only could provide an important resource for further investigating on metabolic pathways in H. serrata, but also shed light on synthetic biology study of HupA.

Determinants of self-efficacy and quality of life in patients with temporary enterostomy: a cross-sectional survey.

AIMS AND OBJECTIVES: To identify determinants of self-efficacy and quality of life in patients with temporary enterostomy. BACKGROUND: Anterior resection with temporary enterostomy is the preferred treatment for patients with rectal cancer, which may impair patients' quality of life. So far, most studies have focused on quality of life in patients with permanent enterostomy, but few studies have looked at that in those with temporary enterostomy. Self-efficacy may determine quality of life in these patients, but few studies have identified determinants of self-efficacy and quality of life. DESIGN: Multicentre, cross-sectional survey and regression analysis to identify determinants of self-efficacy and quality of life. METHODS: A convenience sample of patients undergoing temporary enterostomy at five hospitals in Guangdong Province (China) were surveyed at least four weeks after stoma surgery using validated Chinese versions of internationally recognised questionnaires, including a Stoma Self-Efficacy Scale and the City of Hope Quality of Life-Ostomy Questionnaire. Backward multiple regression analysis was performed to identify whether quality of life was determined by self-efficacy and other clinico-demographic characteristics. RESULTS: Of the 180 questionnaires distributed, 149 (82·8%) were returned, and 135 (75%) were used in the final analysis. Mean global quality of life was 5·40 ± 1·58, and mean global self-efficacy was 79·59 ± 20·21. Significant determinants of self-efficacy and quality of life were identified (ß = 0·62, p < 0·01). Quality of life was determined by type of enterostomy (ß = 0·18, p = 0·01) and payment method (ß = 0·14, p = 0·03). CONCLUSIONS: Quality of life may be determined by self-efficacy, type of enterostomy and payment method, after temporary enterostomy. RELEVANCE TO CLINICAL PRACTICE: Promoting stoma-related self-efficacy in patients with temporary enterostomy may improve their quality of life. Healthcare providers should focus on quality of life in those either with temporary loop ileostomy or entirely self-funded for medicine.

Self-efficacy and Associated Factors in Patients With Temporary Ostomies: A Cross-sectional Survey.

PURPOSE: To examine stoma self-efficacy (SE) and its association with health-related quality of life (HRQOL) and social support in patients with temporary ostomies. SUBJECTS AND SETTING: Convenience sampling was used to recruit 150 patients from 5 hospitals in Guangdong province, China, who had been living with a temporary ostomy for at least 1 month. DESIGN: Cross-sectional survey. METHODS: Respondents completed a questionnaire that included ostomy-related sociodemographic and clinical data, and Chinese language versions of several validated instruments, the Stoma Self-efficacy Scale (C-SSES), City of Hope-Quality of Life-Ostomy Questionnaire (C-COH-QOL-OQ), and Perceived Social Support Scale (C-PSSS). RESULTS: Of the 150 questionnaires distributed, 122 (81.3%) were returned, and 111 (74%) had sufficiently complete responses to be included in the final analysis. The average score from the C-SSES was 78.55 ± 14.72 (mean ± standard deviation) for total stoma SE; 85.6% of respondents showed low or moderate self-efficacy related to ostomy care. The scores from the C-SSES were 39.36 ± 7.72 for stoma care SE and 23.33 ± 6.69 for social SE. Stoma care SE was significantly associated with HRQOL domains, psychological well-being (B = 2.09, P < .01), social well-being (B = 1.16, P < .05), significant other support (B = 1.19, P < .01), and friend support (B = 0.72, P < .01). Social SE was associated with education level (B = 2.84, P < .01), HRQOL domains, psychological well-being (B = 1.88, P < .01), social well-being (B = 1.17, P < .01), and family support (B = 0.48, P < .01). CONCLUSIONS: Persons with temporary ostomies reported low or moderate levels of SE, suggesting the need to focus on HRQOL aspects of psychological and social well-being, as well as social support. We hypothesize that interaction with other ostomy patients, especially those with long-term enterostomy experience or those trained through ostomy organizations, may improve stoma SE.

An efficient in vitro organogenesis protocol for the endangered relic tree species Bretschneidera sinensis and genetic fidelity assessment using DNA markers.

Bretschneidera sinensis is a monotypic species of rare and tertiary relic trees mainly distributed in China. B. sinensis is a potentially valuable horticultural plant, which has significant ornamental and research value, and is a crucial tool for the study of phylogeography. The artificial cultivation of B. sinensis is of great scientific value and practical significance. In this study, we developed a direct organogenesis process of B. sinensis using mature zygotic embryos as initial materials. The highest sterile germination induction (54.5%) from the mature zygotic embryo was obtained in a Murashige and Skoog (MS) medium with 2.0 mg·L-1 6-benzylaminopurine (6-BA) and 0.2 mg·L-1 α-naphthaleneacetic acid (NAA). The highest percentage of shoot regeneration (90.37%) was attained using 1.0 mg·L-1 6-BA and 0.01 mg·L-1 NAA in the MS medium. The Woody Plant Medium (WPM) had the greatest adventitious shoot elongation rate of 93.33%. The most optimized rooting rate was 88.89% in a half-strength MS medium containing 2.0 mg·L-1 indole-3-butyric acid (IBA) and 1.0 mg·L-1 NAA. The genetic fidelity of in vitro regenerated plantlets was assessed using inter-simple sequence repeats and random amplified polymorphic DNA molecular markers, confirming the genetic uniformity and stability of regenerated B. sinensis plantlets. Our research presents an effective in vitro propagation system for B. sinensis, laying the groundwork for its germplasm conservation and large-scale production while maintaining high genetic integrity.

A Novel Strategy for Whole-Cell Biotransformation Enabling Simultaneous l-Phenyllactic Acid Production and Coenzyme Regeneration.

l-Phenyllactic acid (l-PLA) is a small molecular organic acid that exhibits a powerful capacity for inhibition against foodborne pathogens. In this work, we developed a new cost-effective and environmentally friendly process for the biosynthesis of l-PLA. This strategy designed a novel whole-cell biotransformation system employing two heterologous enzymes, namely, phenylalanine dehydrogenase (PheDH) and l-hydroxyisocaproate dehydrogenase (l-HicDH). The novelty of this strategy lies in the first-time utilization of these two enzymes, which not only enables cascade catalysis for the production of l-PLA but also facilitates the regeneration of the coenzymes (NAD+/NADH) using only two enzymes rather than introducing more heterologous enzymes to the system. Consequently, this strategy can effectively simplify the biosynthesis process of l-PLA and minimize production costs. The initial l-PLA yield using this process achieved 2.53 ± 0.07 g/L. Furthermore, through meticulous optimization of the parameters for inducible enzyme expression and l-PLA biosynthesis, the l-PLA yield was successfully increased to 4.68 ± 0.04 g/L with a yield rate of 64.54 ± 0.29%. Moreover, this novel strategy is versatile in the biosynthesis of other organic acids, which can be achieved by easily modulating the combinations of substrates and enzymes.

Salt stress downregulates 2-hydroxybutyrylation in Arabidopsis siliques.

Lysine 2-hydroxyisobutyrylation (Khib) is one of the newly discovered post-translational modifications (PTMs) through protein acylation. It has been reported to be widely distributed in both eukaryotes and prokaryotes, and plays an important role in chromatin conformation change, gene transcription, protein subcellular localization, protein-protein interaction, signal transduction, and cellular proliferation. In this study, the Khib modification proteome of siliques from A. thaliana under salt stress (Ss) and those in the control (Cs) were compared. The results showed that Khib modification was abundant in siliques. Totally 3810 normalized Khib sites on 1254 proteins were identified, and the Khib modification showed a downregulation trend dramatically: it was down-regulated at 282 sites on 205 proteins while was up-regulated at 96 sites on 78 proteins in Ss siliques (Data are available via ProteomeXchange with identifier PXD028116 and PXD026643). Among them, 13 proteins, including F4IVN6, Q9M1P5, and Q9LF33, had sites with the most significant regulation of Khib modification. Bioinformatics analysis suggested that the differentially Khib-regulated proteins mainly participated in glycolysis/gluconeogenesis and endocytosis. In particular, there were differentially117 Khib-regulated proteins that were mapped to the protein-protein interaction database. In the KEGG pathway enrichment analysis, Khib-modified proteins were enriched in several pathways related to energy metabolism, including gluconeogenesis pathway, pentose phosphate pathway, and pyruvate metabolism. Overall, our work reveals the first systematic analysis of Khib proteome in Arabidopsis siliques under salt stress, and sheds a light on the future studies on the regulatory mechanisms of Khib during the salt stress response of plants. SIGNIFICANCE: In this study, we found the Khib-modified proteins in silique under salt stress and described the enrichment of Khib-modified proteins involved in the biological processes and cellular localization. Proteins undergoing 2-hydroxyisobutylation were mainly involved in the gluconeogenesis pathway, pentose phosphate pathway, and pyruvate metabolism, suggesting that 2-hydroxyisobutylation affects the energy metabolic pathway, and thus the development of the plant. In addition, specific candidate proteins that may affect plant development under salt stress were selected. This study will provide a theoretical basis for revealing the function and mechanism of these proteins and their 2-hydroxyisobutyryl modifications during the development of silique under salt stress.

Relationship among self-concept clarity, social support, and psychological resilience in Chinese patients with an enterostomy: A cross-sectional study and structural equation model analysis.

PURPOSE: This study was to explore the relationship between self-concept clarity, social support, and psychological resilience in Chinese enterostomy patients. METHODS: A cross-sectional, descriptive study was conducted from October 2019 to April 2020 among enterostomy patients visiting the stoma clinic and inpatients with an enterostomy in general surgery of three tertiary grade A hospitals in Guangzhou, China. Patients (n=201) were required to complete paper questionnaires, including sociodemographic information, the 10-Item Resilience Scale Specific to Cancer, Self-concept Clarity Scale, and Social Support Rating Scale. Multiple linear regression analyses and structural equation modeling were conducted to explore the effect of self-concept clarity and social support on psychological resilience. RESULTS: Psychological resilience was at a median level among enterostomy patients, with a mean score of 38.21 (SD, 7.24). Education level (ß = 1.032, P = .004), own acceptance of stoma (ß = 2.445, P < .001), social support (ß = 206, P = .001) and self-concept clarity (ß = 0.285, P < .001) were factors related to psychological resilience, explaining 33.1% of the variance of psychological resilience. It was also shown that self-concept clarity exerted its direct positive effect on social support (ß = 0.098, SE = 0.025, BC 95%CI = 0.047/0.143), and it also had an indirect positive effect on psychological resilience through mediating social support (ß = 0.193, SE = 0.033, BC 95%CI = 0.121/0.245). CONCLUSIONS: This study suggests that self-concept clarity has an indirect positive effect on psychological resilience by influencing social support in enterostomy patients. Medical staff should strive to improve the self-concept clarity as a new means to enhance patients' psychological resilience, especially focusing on patients' social support, education level and own acceptance of enterostomy.

Effects of Evidence-Based Continuing Care Bundle on Health Outcomes in Rectal Cancer Patients With Temporary Stomas: A Multicenter Randomized Controlled Trial.

BACKGROUND: The patients with temporary stomas after anterior resection for rectal cancer may experience significant impact on their health outcomes, and hence continuing care is necessary and important for these patients. However, the effects of some single continuing care interventions remain unclear. Continuing care bundle may be an effective approach to address this uncertainty. OBJECTIVE: The aim of this study was to investigate the effects of an evidence-based continuing care bundle on selected health outcomes in patients with temporary stomas after anterior resection for rectal cancer. METHODS: This was a multicenter randomized controlled trial. A total of 124 patients with temporary stomas after anterior resection for rectal cancer were recruited from 4 general tertiary hospitals in Guangzhou, China, and were randomly assigned to a control group or an intervention group. Both groups received usual care, whereas the intervention group additionally received evidence-based continuing care bundle. Self-efficacy, quality of life, and stoma-related complications were collected at baseline and 4 and 12 weeks after surgery. Satisfaction and outcomes of stoma reversal were collected at the end of the observation. RESULTS: The intervention group had significantly improved the self-efficacy (F = 11.88, P = .001), quality of life (F = 17.99, P < .001) over time, satisfaction (t = 4.08, P < .001), and outcomes of stoma reversal (χ2 = 5.93, P = .015) and reduced the incidence of complications (P < .05). CONCLUSIONS: Evidence-based continuing care bundle can be an effective method to improve the health outcomes among these patients. IMPLICATION FOR PRACTICE: By using the evidence-based continuing care bundle, nurses can help these patients improve their health outcomes in stoma-specific nursing.

Efficient Tissue Culture Protocol for Magnolia lucida (Magnoliaceae) and Confirmation of Genetic Stability of the Regenerated Plants.

Magnolia lucida (Magnoliaceae) is classified as an endangered species by the International Union for Conservation of Nature. It has high commercial value owing to its attractive tree shape and flowers. We adopted an excellent genotype of M. lucida as the parent material and established a mini-cut orchard through grafting to provide trunk shoots explants over the long-term. Optimal sterilization was achieved using a combination of 75% ethanol for 30 s, one percent benzalkonium bromide for five minutes, and 0.1% mercuric chloride for five minutes. Modified Murashige and Skoog medium (ML) was the optimal medium for the growth of M. lucida. Addition of one mg/L of 6-benzyl adenine (BA) and 0.05 mg/L of α-naphthaleneacetic acid (NAA) to the medium increased the shoot induction rate to 95.56%, and the ML medium containing 0.4 mg/L BA and 0.04 mg/L NAA achieved the maximum multiplication rate (284.56%). Dark treatment for seven days, followed by continuous light treatment could better resolve the challenge of difficult rooting in M. lucida plants. Using random amplified polymorphic DNA and inter simple sequence repeat markers, we confirmed the genetic uniformity and stability of the regenerated plants. Our protocol should be helpful for the propagation and conservation of this endangered plant.

OsHLH61-OsbHLH96 influences rice defense to brown planthopper through regulating the pathogen-related genes.

BACKGROUND: In plants, basic helix-loop-helix (bHLH) proteins form the largest transcription factor (TF) family. Among them, HLH proteins are a small group of atypical members that lack the basic domain, and form dimers with bHLH proteins. Although bHLH proteins have been proved to play important roles in plant development and physiology, the function of HLH proteins is rarely studied, not to mention in plant biotic resistance. Brown planthopper (BPH) is a kind of rice-specific insect that causes devastating yield losses each year. RESULTS: In this study, we identified OsHLH61 gene that encodes HLH protein. OsHLH61 gene could be highly induced by BPH infestation. Furthermore, Methyl Jasmonic acid (Me-JA) and cis-12-oxo- phytodienoic acid (OPDA) induced expression of OsHLH61, while SA repressed it. We knocked down expression of OsHLH61 by RNA interference (RNAi), the transgenic plants were susceptible to BPH infestation. RNA-seq analysis revealed that some pathogen-related (PR) genes in the Salicylic acid (SA) signaling pathway that mediate plant immunity were obviously down-regulated in the OsHLH61 RNAi plants. Meanwhile, yeast two-hybrid assay and bimolecular luciferase complementation (BiLC) analysis identified bHLH096 to be an interacting factor of OsHLH61. Also, some PR genes were down-regulated in the OsbHLH96 over expressing lines. Expression of OsbHLH96 was inhibited. Besides, OsbHLH96 might interact with Jasmonate Zim-Domain3 (OsJAZ3). CONCLUSION: Altogether, we identified an OsHLH61-OsbHLH96 complex that might mediate defense to BPH through regulating PR genes. And OsHLH61-OsbHLH96 might be important in mediating SA and JA signaling crosstalk.

An efficient micropropagation protocol for an endangered ornamental tree species (Magnolia sirindhorniae Noot. & Chalermglin) and assessment of genetic uniformity through DNA markers.

Magnolia sirindhorniae Noot. & Chalermglin is an endangered species with high ornamental and commercial value that needs to be urgently protected and judiciously commercialized. In this study, a protocol for efficient regeneration of this species is standardized. The lateral buds of the M. sirindhorniae plant were used as an explant. Half-strength Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 6-benzyladenine (BA), 0.1 mg/L α-naphthaleneacetic acid (NAA), and 2.0 mg/L gibberellic acid (GA3) was found to be the optimal medium for shoot induction. The maximum shoot multiplication rate (310%) was obtained on Douglas-fir cotyledon revised medium (DCR) fortified with 0.2 mg/L BA, 0.01 mg/L NAA, and additives. The half-strength DCR medium supplemented with 0.5 mg/L NAA and 0.5 mg/L indole-3-butyric acid (IBA) supported the maximum rate (85.0%) of in vitro root induction. After a simple acclimatization process, the survival rate of plantlets in a substrate mixture of sterile perlite and peat soil (1:3; v/v) was 90.2%. DNA markers were used for assessment of genetic uniformity, confirming the genetic uniformity and stability of regenerated plants of M. sirindhorniae. Thus, the described protocol can safely be applied for large scale propagation of this imperative plant.

The Adsorption of Cd(II) on Manganese Oxide Investigated by Batch and Modeling Techniques.

Manganese (Mn) oxide is a ubiquitous metal oxide in sub-environments. The adsorption of Cd(II) on Mn oxide as function of adsorption time, pH, ionic strength, temperature, and initial Cd(II) concentration was investigated by batch techniques. The adsorption kinetics showed that the adsorption of Cd(II) on Mn oxide can be satisfactorily simulated by pseudo-second-order kinetic model with high correlation coefficients (R² > 0.999). The adsorption of Cd(II) on Mn oxide significantly decreased with increasing ionic strength at pH < 5.0, whereas Cd(II) adsorption was independent of ionic strength at pH > 6.0, which indicated that outer-sphere and inner-sphere surface complexation dominated the adsorption of Cd(II) on Mn oxide at pH < 5.0 and pH > 6.0, respectively. The maximum adsorption capacity of Mn oxide for Cd(II) calculated from Langmuir model was 104.17 mg/g at pH 6.0 and 298 K. The thermodynamic parameters showed that the adsorption of Cd(II) on Mn oxide was an endothermic and spontaneous process. According to the results of surface complexation modeling, the adsorption of Cd(II) on Mn oxide can be satisfactorily simulated by ion exchange sites (X2Cd) at low pH and inner-sphere surface complexation sites (SOCd⁺ and (SO)2CdOH- species) at high pH conditions. The finding presented herein plays an important role in understanding the fate and transport of heavy metals at the water-mineral interface.

OsMADS1 Represses microRNA172 in Elongation of Palea/Lemma Development in Rice.

Specification of floral organ identity is critical for the establishment of floral morphology and inflorescence architecture. Although multiple genes are involved in the regulation of floral organogenesis, our understanding of the underlying regulating network is still fragmentary. MADs-box genes are principle members in the ABCDE model that characterized floral organs. OsMADS1 specifies the determinacy of spikelet meristem and lemma/palea identity in rice. However, the pathway through which OsMADS1 regulates floral organs remains elusive; here, we identified the microRNA172 (miR172) family as possible regulators downstream of OsMADS1. Genetic study revealed that overexpression of each miR172 gene resulted in elongated lemma/palea and indeterminacy of the floret, which resemble the phenotype of osmads1 mutant. On the contrary, overexpression of each target APETALA2 (AP2) genes resulted in shortened palea/lemma. Expression level and specificity of miR172 was greatly influenced by OsMADS1, as revealed by Northern blot analysis and In situ hybridization. Genetically, AP2-3 and AP2-2 over expression rescued the elongation and inconsistent development of the lemma/palea in OsMADS1RNAi transgenic plants. Our results suggested that in rice, OsMADS1 and miR172s/AP2s formed a regulatory network involved in floral organ development, particularly the elongation of the lemma and the palea.