Identification, expression and protein interaction analysis of Aux/IAA and ARF gene family in Senna tora L / 药学学报

The early response of plant auxin gene family Aux/IAA (auxin/indole-3-acetic acid) and its interaction with auxin response factor (ARF) are important pattern to regulate plant growth and development. This work identified 28 StoIAA and 24 StoARF members based on the whole genome data of the medicinal plant Senna tora L., which were classified into 10 and 8 subfamilies, respectively. Phylogenetic tree and collinearity analysis showed that S. tora has close evolutionary relationship with the IAA and ARF homologous genes of Glycine max, Medicago truncatula, and the segment duplication events dominate the expansion of StoIAA and StoARF. Gene structure analysis showed that the vast majority of StoIAA and StoARF contain characteristic conserved domain. Transcriptome data showed that StoIAAs and StoARFs were expressed in leaves, roots and seeds, some members had tissue specific expression. The StoIAA and StoARF promoter region most contain functional elements related to stress response, growth and development, hormone induction and secondary metabolism. In addition, gene expression analysis showed that many StoIAAs and StoARFs can quickly respond to drought and salt stress and exhibited same expression patterns under both stress condition. The yeast two-hybrid experiment confirmed that StoARF8 and StoARF10 exhibit varying degrees of interaction with multiple StoIAA proteins, respectively. The above results provide a basis for further biological functional analysis of the Aux/IAA and ARF gene family of S. tora.

Ornamental plant Pachystachys lutea as a source of promising endophytes for plant growth and phytoprotective activity

Endophytes are growth-promoting agents capable of synthesizing phytohormones, uptaking nutrients, and controlling pathogens. There is a strong potential to exploit them in the agriculture field like biofertilizers and biocontrol agents. In this work, we aimed to evaluate endophytic fungi isolated from Pachystachys lutea for their potential to solubilize phosphate, synthesise indole acetic acid (IAA), antagonize phytopathogens, and promote plant growth under greenhouse conditions. The phosphate solubilization efficiency was assessed on Pikovskaya’s agar medium. For analysis of IAA production, mycelia plugs of endophytes were cultured in Potato Dextrose Broth medium supplemented with L-tryptophan, with Salkowski Reagent, and the absorbance of the culture was measured. The antagonism evaluation of strain Alternaria sp. PL75 against phytopathogens was performed using the paired-culture technique. The promotion of plant growth provided by Alternaria sp. PL75 was evaluated in tomato plants. All strains evaluated were able to solubilize phosphate; however, the strain Alternaria sp. PL75 was the most effective (4.29). Two strains, Nemania sp. PL27 and Alternaria sp. PL75, produced 1.86 and 1.73 & 956;g mL-1 of IAA, respectively. In the antagonism assay, the endophyte Alternaria sp. PL75 and its fungal extract showed the best results against the pathogen Moniliophthora perniciosa. The greenhouse experiment result showed the endophyte Alternaria sp. PL75 increased the plantlets emergency speed index and the percentage of germination from 60 to 81.63%. It was also observed a statistical significance in the shoot length of the treated plants with the endophyte suspension (55.38 cm) compared to the control (41.67 cm).

Screening and evaluation of PGPR strains having multiple PGP traits from hilly terrain

Plant growth-promoting rhizobacteria (PGPR) with multiple beneficial traits serve as potentially, ecofriendly,and cost-effective alternatives to chemical fertilizers and pesticides. They have both direct and indirectaffirmative impacts on overall plant growth and health. PGPRs are well known to directly improve the plantgrowth by phytohormone production and availability of minerals in soil. A total of nine soil samples were takenfrom near the rhizospheric zone of different crops and 56 rhizobacterial strains were isolated. Only 16 out of56 rhizobacterial strains were found positive for more than one beneficial trait that included solubilization ofphosphate, indole acetic acid (IAA), siderophore, ammonia and H2S production. Among all PGPR strains,RKM15 was observed having the highest phosphate solubilizing index (3.4), solubilized phosphorus (339mg L−1) and also siderophore unit (70.54 %). The maximum IAA production was observed by RKM25 strain(35.56 µg ml−1). The most promising RKM15 isolate was identified as Pantoea dispersa (MN629239) through16S rRNA gene sequencing technique. This characterized PGPR strain may be used for the development ofbiofertilizers to enhance crop productivity and improvement in soil fertility

Clinical characteristics of insulin autoimmune syndrome caused by methimazole in Chinese population / 中国临床药理学与治疗学

AIM: To explore the clinical features of insulin autoimmune syndrome (IAS) induced by methimazole in Chinese population. METHODS: The literature on IAS cases caused by methimazole in the Chinese population published before and after June 30, 2019 was collected for retrospective analysis.RESULTS:The age of onset of men was earlier than that of women, and the sex ratio was 1:2.31 in 95 patients.After taking methimazole 30 mg/d, IAS occurred most from one month to three months,characterized by neuropathic hypoglycemia as the first symptoms at night and early in the morning,blood sugar below 2 mmol/L,insulin concentration≥100 mU/L, IAA positive and no obvious abnormalities in pancreas imaging.The symptoms gradually relieved after symptomatic treatment and stopped taking methimazole. There was no significant difference in the time of hypoglycemia disappearance between the 54 patients who received hormone therapy and non-hormone therapy.CONCLUSION:Methimazole-induced IAS is a clinically rare autoimmune disease. It should be treated promptly occuring hypoglycemia or hyperglycemia during medication.The prognosis of IAS is generally good after proper treatment.

Effect of Rhustox on Micropropagation of Scoparia dulcis L. through Leaf Explants Culture

Aim: To investigate the effect of Rhus toxicodendron (30CH) along with different compositions of phytohormones (Auxin and Cytokinin) on the basis of growth and multiplication of explants under optimum temperature under in-vitro conditions. Study Design: To establish and design the standard protocol for the in-vitro propagation through leaf explant of Scoparia dulcis under stress of phytohormones and homeopathic medicine Rhus toxicodendron (30CH). Place and Duration of Study: The plant materials were procured from the Herbal Botanical Garden Patna Science College, Department of Botany, Patna University, Patna, Bihar. The experimental part was carried out in Plant Tissue Culture Laboratory, between December 2017 to August 2018 in Department of Botany P.U. Patna. Methodlogy: The sterilized leaf explants were inoculated into MS media fortified with different phytohormones (Auxin and Cytokinin) and Rhus tox(30CH) under aseptic environmental conditions for the growth and development of callus, embryoids etc. Result: The explants in MS medium supplemented with auxins phytohormones and Rhus tox(30CH) exhibited that IAA (0.10 to 2.0 mg/l) and BAP (0.10 to 2.5 mg/l) induces green and compact calli. Whereas at 0.30mg/l of IAA and 0.50 mg/l BAP induced brown friable calli. 2,4-D (1.5 mg/l) and Kinetin (1.5-6.5mg/l) concentrations induced brown and friable calli. Rhus tox(30CH) (100 µl/100 ml) enhances proliferation with 2,4-D and Kinetin (1.5/1.5 mg/l.). Conclusion: After 42 days of culture initiation and establishment the callus was 520.0±1.12 mg in the mixture of 2,4-D and Kinetin (1.5 mg/l) in Rhus tox free medium. Whereas weight of callus were found to be 1092±0.74 mg after 42 days in the same medium of 2, 4-D and Kinetin (1.5/5.5 mg/l) supplemented with Rhus tox (100 µl/100 ml). Hence, the investigation proponded that the Rhus tox (CH30) has increased the rate of callus development and plantlet regeneration.

Functional Attributes and Antagonism of Root-Associated Bacteria of Different Rice Cultivars from Acid Soil

Abstract: This study evaluated root endophyte bacteria and rhizobacteria in terms ofmultifaceted plant growth promotion (PGP) traits and antagonistic potential against majorfungal pathogens of rice (viz. Rhizoctonia solani, Bipolaris oryzae, Pyricularia oryzae,Ustilaginoidea virens and Sarocladium oryzae). Twenty bacterial isolates from each group(viz. endorhiza and rhizosphere) were isolated from different rice types of North EastIndia. Cultivated rice types were Upland rice (Mima, Kochi and Minil), Lowland HYV rice(Ranjit and Jaya) and Lowland scented rice (Keteki Joha and Kunkuni Joha). Thepopulation of rhizospheric bacteria was higher than the endophyte isolates. Thepopulation counts of endophyte bacteria were the highest in scented rice cultivar(Kunkuni Joha) and for rhizobacterial isolates; the population density was maximum inthe lowland HYV rice (Jaya). The endophytes were more efficient than the rhizobacteria interms of the activity of cellulase, pectinase, ACC-deaminase, production of IAA- likesubstances, solubilization of zinc and mineralization of organic phosphates. In contrast,the rhizobacterial isolates were more efficient in solubilization of inorganic phosphatesand antagonism against major rice fungal pathogens. Through 16S rDNA sequenceanalysis, the promising rhizobacterial isolates showing antagonism against all the fungalpathogens were identified as Brevibacillus reuszeri 12R, Lysinibacillus xylanticus 48R,Bacillus megaterium 58R and Serratia marcescens 79R. These results suggest that the rootbacterial endophytes and rhizobacteria characterized in this study could be successfullyused to promote plant growth and induce fungal resistance in rice plants and can be usedas bioinoculants for enhancing rice growth in the acid soil regions.

Characterization of indole-3-acetic acid in cambial zone tissue by ultra-performance liquid chromatography-tandem mass spectrometry

The developmental plasticity of plants according to changes in their growth conditions is mediated by signalingmolecules called hormones. The major classes of plant hormones are auxin, gibberellins, abscisic acid, andethylene. Among these, the most important is indole-3-acetic acid (IAA). Quantification of IAA in an extract ofapproximately 10 mg obtained from the cambial zone of Eucalyptus leaves was performed by ultra-performanceliquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC/ESI-MS/MS). Theleaves were extracted using acetone, and the extract was analyzed using a reverse-phase column (Acquity UPLCBEH C18, 2.1 mm × 100 mm, 1.7 µm) at a flow rate of 0.2 mL/min with gradient elution of an aqueous mobile phase(containing 0.1% formic acid) with methanol. This gradient elution provided an excellent performance in terms ofspecificity/selectivity, linearity, precision, and ruggedness/stability. In addition, the run time was short (6 min), withexcellent linearity (R2 > 0.99) in the range of 10–70 ng/mL. The structure of IAA was elucidated using UPLC/ESIMS/MS, operating and quantifying in multiple reaction monitoring (MRM) mode.

Mangrove endophyte promotes reforestation tree (Acacia polyphylla) growth

ABSTRACT Mangroves are ecosystems located in the transition zone between land and sea that serve as a potential source of biotechnological resources. Brazil's extensive coast contains one of the largest mangrove forests in the world (encompassing an area of 25,000 km2 along all the coast). Endophytic bacteria were isolated from the following three plant species: Rhizophora mangle, Laguncularia racemosa and Avicennia nitida. A large number of these isolates, 115 in total, were evaluated for their ability to fix nitrogen and solubilize phosphorous. Bacteria that tested positive for both of these tests were examined further to determine their level of indole acetic acid production. Two strains with high indole acetic acid production were selected for use as inoculants for reforestation trees, and then the growth of the plants was evaluated under field conditions. The bacterium Pseudomonas fluorescens (strain MCR1.10) had a low phosphorus solubilization index, while this index was higher in the other strain used, Enterobacter sp. (strain MCR1.48). We used the reforestation tree Acacia polyphylla. The results indicate that inoculation with the MCR1.48 endophyte increases Acacia polyphylla shoot dry mass, demonstrating that this strain effectively promotes the plant's growth and fitness, which can be used in the seedling production of this tree. Therefore, we successfully screened the biotechnological potential of endophyte isolates from mangrove, with a focus on plant growth promotion, and selected a strain able to provide limited nutrients and hormones for in plant growth.

Azotobacter chroococcum as a potentially useful bacterial biofertilizer for cotton (Gossypium hirsutum): Effect in reducing N fertilization / Azotobacter chroococcum como biofertilizante bacteriano potencialmente útil para el algodón (Gossypium hirsutum L.): efecto en la reducción de la fertilización nitrogenada

The aim of this research was to evaluate whether the application of two plant growth-promoting (rhizo)bacteria might reduce nitrogen fertilization doses in cotton. We used strains Azotobacter chroococcum AC1 and AC10 for their proven ability to promote seed germination and cotton growth. These microorganisms were characterized by their plant growth-promoting activities. Then, we conducted a glasshouse study to evaluate the plant growth promoting ability of these strains with reduced doses of urea fertilization in cotton. Results revealed that both strains are capable of fixing nitrogen, solubilizing phosphorus, synthesizing indole compounds and producing hydrolytic enzymes. After 12 weeks, the glasshouse experiment showed that cotton growth was positively influenced due to bacterial inoculation with respect to chemical fertilization. Notably, we observed that microbial inoculation further influenced plant biomass (p<0.05) than nitrogen content. Co-inoculation, interestingly, exhibited a greater beneficial effect on plant growth parameters compared to single inoculation. Moreover, similar results without significant statistical differences were observed among bacterial co-inoculation plus 50% urea and 100% fertilization. These findings suggest that coinoculation of A. chroococcum strains allow to reduce nitrogen fertilization doses up to 50% on cotton growth. Our results showed that inoculation with AC1 and AC10 represents a viable alternative to improve cotton growth while decreasing the N fertilizer dose and allows to alleviate the environmental deterioration related to N pollution.

El objetivo de esta investigación fue evaluar si la aplicación de 2 (rizo)bacterias promotoras del crecimiento vegetal podría reducir la dosis de fertilizante nitrogenado en el cultivo de algodón. Se usaron las cepas Azotobacter chroococcum AC1 y AC10 por su habilidad para promover la germinación de semillas y el crecimiento del algodonero. Estos microorganismos fueron caracterizados sobre la base de sus actividades de promoción del crecimiento vegetal. Luego se realizó un estudio de invernadero con plantas de algodón para evaluar la capacidad de promoción del crecimiento vegetal de dichas cepas con dosis reducidas de urea. Los resultados revelaron que ambas cepas son capaces de fijar nitrógeno, solubilizar fósforo, sintetizar compuestos indólicos y producir enzimas hidrolíticas. Después de 12 semanas, el experimento de invernadero permitió observar que el crecimiento del algodón fue influido positivamente por la inoculación bacteriana con respecto a la fertilización química. En particular, se evidenció que la inoculación microbiana impactó más en la biomasa vegetal (p<0,05) que en el contenido de nitrógeno. Curiosamente, la coinoculación exhibió un mayor efecto positivo sobre los parámetros de crecimiento en comparación con la inoculación simple. Además, se observaron resultados similares, sin diferencias estadísticamente significativas, entre la coinoculación bacteriana más del 50% de urea y el 100% de fertilización. Estos hallazgos indican que la coinoculación de las cepas de A. chroococcum AC1 y AC10 permitiría reducir las dosis de fertilización nitrogenada del cultivo de arroz en hasta el 50% y aliviar, de esta manera, el deterioro ambiental relacionado con la contaminación por N.

Assessment of Rice Associated Bacterial Ability to Enhance Rice Seed Germination and Rice Growth Promotion

ABSTRACT The application of beneficial bacteria has recently been used for sustainable agriculture. In current research, 71 bacterial isolates were obtained from rice plant and the rhizosphere soil of different paddy fields in Guilan province, Iran. After primitive investigation, 40 bacteria with typical predominant characteristics were selected. By PCR-RFLP of their 16S r-DNA gene, 8 Operational Taxonomic Units (OTUs) totally consisted of 33 isolates were obtained. From all of them, 8 isolates were selected for rice seed germination experiment, then, effective isolates were used for pot experiment to evaluate their ability for promoting rice growth. All of them were able to increase rice growth and yield, but in different potential. These tested isolates were identified as Alcaligenes faecalis (DEp8, O1R4), Pantoea ananatis (AEn1), Bacillus vietnamensis (MR5), Bacillus idriensis (MR2) and Stenotrophomonas maltophilia by partial sequencing of their 16S r-DNA gene. Among them, AEn1 and MR5 produced indole-3- acetic acid (IAA) in larger amounts than the other isolates and the isolates AEn1 and O1R4 were able to solubilize phosphate in higher amounts. According to the results obtained, it can be concluded that AEn1, O1R4 and MR5 can be considered as bacterial inoculants to use as alternatives for chemical fertilizers.

Diagnostic Value of Combined Detection of IAA, ICA and GADA in the Classification of Diabetes Mellitus / 现代生物医学进展

Objective:To study the diagnostic value of combined detection of IAA,ICA and GADA in the classification of diabetes mellitus.Methods:30 cases of patients with type 1 diabetes who were treated in our hospital from June 2015 to June 2016 were selected as A group,60 cases of patients with type 2 diabetes were selected as B group,50 cases of healthy people were selected as C group.The IAA,ICA and GADA of the three groups were detected by ELISA,and the positive rate of the three groups were compared.Results:The fasting glucose of A group was (10.12± 3.68) mmol/L,B group was (11.23± 3.26) mmol/L,A group and B group were significantly higher than that of C group (P＜0.05),but there was no significant difference between A group and B group (P＞0.05);the positive rates of GADA,ICA and IAA in A group and B group were significantly higher than those in C group (P＜0.05),and the positive rates of GADA,ICA and IAA in A group were significantly higher than those in B group (P＜0.05);the sensitivity and specificity of combined detection of IAA,ICA and GADA in type 2 and type 1 diabetes mellitus were significantly higher than that in the single test (P＜0.05).Conclusions:The combined detection of IAA,ICA and GADA has a high diagnostic value in the classification of diabetes mellitus,which is worth clinical application.

Soil bacteria showing a potential of chlorpyrifos degradation and plant growth enhancement

ABSTRACT Background: Since 1960s, the organophosphate pesticide chlorpyrifos has been widely used for the purpose of pest control. However, given its persistence and toxicity towards life forms, the elimination of chlorpyrifos from contaminated sites has become an urgent issue. For this process bioremediation is the method of choice. Results: Two bacterial strains, JCp4 and FCp1, exhibiting chlorpyrifos-degradation potential were isolated from pesticide contaminated agricultural fields. These isolates were able to degrade 84.4% and 78.6% of the initial concentration of chlorpyrifos (100 mg L-1) within a period of only 10 days. Based on 16S rRNA sequence analysis, these strains were identified as Achromobacter xylosoxidans (JCp4) and Ochrobactrum sp. (FCp1). These strains exhibited the ability to degrade chlorpyrifos in sterilized as well as non-sterilized soils, and were able to degrade 93-100% of the input concentration (200 mg kg-1) within 42 days. The rate of degradation in inoculated soils ranged from 4.40 to 4.76 mg-1 kg-1 d-1 with rate constants varying between 0.047 and 0.069 d-1. These strains also displayed substantial plant growth promoting traits such as phosphate solubilization, indole acetic acid production and ammonia production both in absence as well as in the presence of chlorpyrifos. However, presence of chlorpyrifos (100 and 200 mg L-1) was found to have a negative effect on indole acetic acid production and phosphate solubilization with percentage reduction values ranging between 2.65-10.6% and 4.5-17.6%, respectively. Plant growth experiment demonstrated that chlorpyrifos has a negative effect on plant growth and causes a decrease in parameters such as percentage germination, plant height and biomass. Inoculation of soil with chlorpyrifos-degrading strains was found to enhance plant growth significantly in terms of plant length and weight. Moreover, it was noted that these strains degraded chlorpyrifos at an increased rate (5.69 mg-1 kg-1 d-1) in planted soil. Conclusion The results of this study clearly demonstrate that the chlorpyrifos-degrading strains have the potential to develop into promising candidates for raising the productivity of crops in pesticide contaminated soils.

Isolation and identification by 16S rRNA sequence analysis of plant growth-promoting azospirilla from the rhizosphere of wheat

ABSTRACT The main objective of the present study was to isolate phytohormone-producing, phosphate-solubilizing strains of Azospirillum from wheat to be used as inoculants for plant growth promotion. Five Azospirillum strains were isolated from the rhizosphere of field-grown wheat (Triticum aestivum L.), and it was confirmed by BOX-polymerase chain reaction (PCR) that the isolates were different and not re-isolates of the same strain. Sequence analysis of the PCR-amplified 16S rRNA gene indicated that four isolates showed maximum similarity to Azospirillum brasilense and one isolate showed maximum similarity to Azospirillum zeae. This is the first report indicating the presence of an A. zeae like isolate in the wheat rhizosphere in Pakistan. The bacterial isolates were characterized for their plant growth-promoting traits, phosphate solubilization, and indole-3-acetic acid (IAA) production. None of the isolates showed phosphate solubilization activity in the commonly used Pikovskaya medium. However, all strains (except AzoK4) exhibited ability to solubilize tricalcium phosphate (TCP) in modified Pikovskaya medium in which sucrose was replaced by Na-malate, as well as in TCP-supplemented Luria-Bertani (LB) medium. Organic acids, such as acetic, citric, lactic, malic, and succinic acids, were detected in culture supernatants of the tested Azospirillum strains. All strains exhibited ability to produce IAA in the growth medium, except Azospirillum sp. AzoK1. Among the strains tested, the maximum IAA production (30.49 ± 1.04 mg L-1) and phosphate solubilization (105.50 ± 4.93 mg L-1) were shown by a pure culture of Azospirillum sp. AzoK2. In pot experiments, single-strain inocula of Azospirillum sp. AzoK1 and AzoK2 improved wheat plant growth.

PGP potential, abiotic stress tolerance and antifungal activity of Azotobacter strains isolated from paddy soils.

Azotobacter strains were isolated by serial dilution method and colonies were viscous, smooth, glistening, and brown to black colour on Jenson’s N-free agar. Morphological and biochemical tests showed characteristic features of Azotobacter. Further, molecular analyses revealed the presence of different Azotobacter species viz., A. armeniacus, A. chroococcum, A. salinestris, A. tropicalis and A. vinelandii. The isolates were tested for their ability of nitrogen fixation, indole acetic acid (IAA), gibberllic acid production and phosphate solubilization. Four isolates (GVT-1, GVT-2 KOP-11 and SND-4) were efficient in fixation of highest amount of N2 (29.21 μg NmL-1day-1), produced IAA (25.50 µg mL-1), gibberllic acid (17.25 μg 25 mL-1) and formed larger P solubilizing zone (13.4 mm). Some of the Azotobacter strains were produced siderophores, hydrogen cyanide and were positive for ammonia production with respect to antifungal activity of Azotobacter was tested with dual culture method and A. tropicalis inhibited the growth of Fusarium, Aspergillus and Alternaria species. Azotobacter isolates were tested against salt (0-10%), temperature (4-55ºC), pH (5.0-10) and insecticide chloropyrifos (0-3%) tolerance study. Among them, A. chroococcum was found tolerant to a maximum of 6% NaCl with a temperature of 35-45ºC and to a pH up to 8. All the 4 strains showed effective growth against 3% chloropyrifos concentration. The studies revealed that the Azotobacter strains not only produced plant growth promoting substances but are also tolerant to abiotic stresses such as temperature, pH and insecticides.

Indole acetic acid production by fluorescent Pseudomonas spp. from the rhizosphere of Plectranthus amboinicus (Lour.) Spreng. and their variation in extragenic repetitive DNA sequences.

Fluorescent Pseudomonas (FP) is a heterogenous group of growth promoting rhizobacteria that regulate plant growth by releasing secondary metabolic compounds viz., indole acetic acid (IAA), siderophores, ammonia and hydrogen cyanide. In the present study, IAA producing FPs from the rhizosphere of Plectranthus amboinicus were characterized morphologically, biochemically and at the molecular level. Molecular identification of the isolates were carried out using Pseudomonas specific primers. The effect of varying time (24, 48, 72 and 96 h), Trp concentrations (100, 200, 300, 400 and 500 µg.ml-1), temperature (10, 26, 37 and 50±2 °C) and pH (6, 7 and 8) on IAA production by 10 best isolates were studied. Results showed higher IAA production at 72 h incubation, at 300 µg.ml-1 Trp concentration, temperature 26±2 °C and pH 7. TLC with acidified ethyl acetate extract showed that the IAA produced has a similar Rf value to that of the standard IAA. Results of TLC were confirmed by HPLC analysis. Genetic diversity of the isolates was also studied using 40 RAPD and 4 Rep primers. Genetic diversity parameters such as dominance, Shannon index and Simpson index were calculated. Out of 40 RAPD primers tested, 9 (2 OP-D series and 7 OP-E series) were shortlisted for further analysis. Studies using RAPD, ERIC, BOX, REP and GTG5 primers revealed that isolates exhibit significant diversity in repetitive DNA sequences irrespective of the rhizosphere.

Study on relationship among INS genetic polymorphismsms and the occurrence of type 2 diabetes and serum IAA-Ab levels / 重庆医学

Objective To investigate the relationship between polymorphisms of gene promoter region INS 5′UTR single nu-cleotide and type 2 diabetes and serum IAA-Ab levels.Methods By Sequenom MassArray SNP genotyping detection technology, INS 3 pyomter regime single nucleotide polymorphisms (rs689,rs714641 77 and rs3842738)of 497 patients in Chongqing with type 2 diabetes cases(treatment group)and 500 cases(control group)were genotyped and analyzed.IAA-Ab levels in diabetes patients was detected.Theχ2 test statistic was used to analyze the treatment group and control groups.The genotype frequency distribution of IAA-Ab-positive and negative groups SNP was analyzed by non-conditional logistic regression,adjusted for sex,age impact,cal-culated the odds ratio (OR)and 95 % confidence interval(CI ).The polymorphic loci with type 2 diabetes susceptibility and serum GAD-Ab levels was evaluated.Results The genotype frequency distribution of rs689AA,TT and AT was 58.75%,28.77% and 12.47%,respectively.The control group are 50.40%,35.60% and 14.00% respectively.The difference was statistically significant (χ2 =3.923,P <0.05).Compared with the genotype of AA,TT genotype can decrease risky of diabetes,with OR values 0.35(95%CI :0.18-1.06).There was significant difference of AA,TT,AT genotypes between IAA-Ab negative and IAA-Ab positive pa-tients (P < 0.05 ).Conclusion INS polymorphisms might be related to the risky of type 2 diabetes and serum IAA-Ab level in chinses population.

Rapid propagation of Paris polyphylla var. chinensis / 中草药

Objective: The roots of Paris polyphylla var. chinensis were used as explants to establish a rapid propagation technique. Methods: The effects of hormone 6-BA, 2,4-D, NAA, IBA, KT, IAA, ceftriaxone, activated carbon, etc on callus induction, differentiation of adventitious bud, and formation of adventitious roots were studied to find out the best culture medium formulation by using MS as basic culture medium. Results: The best medium for callus induction of P. polyphylla var. chinensis was MS + 6-BA 1.0 mg/L + NAA 1.5 mg/L + 2,4-D 3.0 mg/L, and the best medium for differentiation of adventitious bud was MS + 6-BA 3.0 mg/L + IAA 0.3 mg/L + KT 1.0 mg/L + ceftriaxone 300 mg/L, while the best medium for formation of adventitious roots was 1/2 MS + IBA 0.5 mg/L + NAA 0.1 mg/L + activated carbon 0.5%. The tissue cultured seedling was moved to sterile water for 5 d when the first leaf emerged and four adventitious roots appeared, then the seedling was transplanted into the soft soil. The tissue cultured seedling would grow strongly with 100% survival rate. Conclusion: The adventitious bud is inducted and then the root from bare-root seedling is cultured on the basis of obtained callus. Finally the best conditions to foster the tissue cultured seedling of P. polyphylla var. chinensis have been screened.

Screening and optimization of low-cost medium for Pseudomonas putida Rs-198 culture using RSM

The plant growth-promoting rhizobacterial strain Pseudomonas putida Rs-198 was isolated from salinized soils from Xinjiang Province. We optimized the composition of the low-cost medium of P. putida Rs-198 based on its bacterial concentration, as well as its phosphate-dissolving and indole acetic acid (IAA)-producing capabilities using the response surface methodology (RSM), and a mathematical model was developed to show the effect of each medium component and its interactions on phosphate dissolution and IAA production. The model predicted a maximum phosphate concentration in medium containing 63.23 mg/L inorganic phosphate with 49.22 g/L corn flour, 14.63 g/L soybean meal, 2.03 g/L K2HPO4, 0.19 g/L MnSO4 and 5.00 g/L NaCl. The maximum IAA concentration (18.73 mg/L) was predicted in medium containing 52.41 g/L corn flour, 15.82 g/L soybean meal, 2.40 g/L K2HPO4, 0.17 g/L MnSO4 and 5.00 g/L NaCl. These predicted values were also verified through experiments, with a cell density of 10(13) cfu/mL, phosphate dissolution of 64.33 mg/L, and IAA concentration of 18.08 mg/L. The excellent correlation between predicted and measured values of each model justifies the validity of both the response models. The study aims to provide a basis for industrialized fermentation using P. putida Rs-198.

Effect of Different Carbon and Nitrogen Sources on Growth and Indole Acetic Acid Production by Rhizobium Species Isolated from Cluster Bean [Cyamopsis tetragonoloba (L.)].

Five Rhizobium strains (Cb1, Cb2, Cb3, Cb4 and Cb5) were isolated from root nodules of cluster bean [Cyamopsis tetragonoloba (L.)] on yeast extract mannitol agar (YMA) medium. All the five Rhizobium isolates have shown the Indole Acetic Acid( IAA) production in culture medium supplemented with L-tryptophan. The IAA content in culture supernatant was estimated by using the colorimetric method (13). All the five Rhizobium isolates produced maximum amount of IAA in medium supplemented with 2.5mg/ml Ltryptophan concentration. Production of IAA was maximum at 72h of incubation when bacteria reached the stationary phase. The cultural conditions were optimized for maximum IAA production by using different carbon and nitrogen sources as well as changing the incubation period. Glucose and L-asparagine were found to be the best carbon and nitrogen sources, respectively for maximum IAA production. The cell wall affecting agent, penicillin increased the IAA production up to 77.95% in Cb4, 59.52% in Cb5 and 37.84% in Cb3 isolates, over the control. Among the five isolates studied, isolate Cb4 showed better performance in IAA production.

Effects of extraction methods of phenolic compounds from Xanthium strumarium L. and their antioxidant activity / Efeito de métodos de extração de compostos fenólicos de Xanthium strumarium L. e suas atividades antioxidantes

Foram avaliados os efeitos de métodos de extração e de solventes sobre o rendimento de extrato, conteúdo de fenólicos totais, atividade antioxidante, e composição dos compostos fenólicos de Xanthium strumarium L. A atividade antioxidante foi determinada pelo método do radical 2,2-difenil-1-picrilhidrazila (DPPH), e a composição dos compostos fenólicos foi determinada por cromatografia líquida de alta eficiência e por espectrometria de massas com ionização por electrospray. Todos os resultados foram afetados pelo método de extração e, principalmente pelo solvente utilizado, sendo que os melhores resultados foram obtidos com o solvente metanol. Os extratos metanólico e etanólico apresentaram forte ação antioxidante e os principais compostos fenólicos encontrados nos extratos foram os ácidos ferúlico e clorogênico.

The effect of extraction methods and solvents on overall yield, total phenolic content, antioxidant activity, and the composition of the phenolic compounds in Xanthium strumarium extracts were studied. The antioxidant activity was determined by using 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), and the composition of the phenolic compounds was determined by HPLC-DAD and LC/MS. All results were affected by the extraction method, especially by the solvent used, and the best results were obtained with the methanol extract. The methanolic and ethanolic extracts exhibited strong antioxidant activity, and the chlorogenic and ferulic acids were the most abundant phenolic compounds in the extracts.