ABSTRACT
Magnolia biondii Pamp is an important ornamental tree species widely grown and used as a rootstock in the propagation of different Magnolia varieties. In the current studies, anatomical, physiological and endogenous hormones were studied to check the effect of IBA 750 mg/L on the adventitious rooting and to provide theoretical and technical support for the propagation of Magnolia biondii Pamp through stem cuttings. Two thousand stem cuttings were prepared and divided into two groups i.e., IBA treated cuttings and water control. For the evaluation of antioxidant enzyme activities, and endogenous hormones levels, samples were collected on the day of planting and each 5th day and further steps were carried out in the laboratory according to the protocols and proper precautions. For the anatomical observations, samples were collected on the 13th, 15th, and 17th day for IBA treated cuttings while 21st, 23rd, and 25th day for control. Collected samples were preserved in the FAA solution and further observations were carried out in the laboratory. Anatomical observations showed that it took 13 days for the differentiation of root primordia to the appearance of young adventitious roots in IBA treated cuttings, while it took 21 days to develop primordia in the control. Antioxidant enzyme activities involved in ROS were significantly higher in the IBA treated cuttings compared to control. POD showed a peak on the 13th day before the emergence of roots in IBA treated cuttings while it showed a peak on the 21st day in the control. PPO showed a peak on the 21st day in the IBA treated cuttings while it showed a peak on the 29th day in the control. SOD showed a peak on the 17th day in IBA treated cuttings, while it showed a peak on the 25th day in the control. Exogenous application of IBA enhanced the endogenous IAA and GA3 levels compared to CK, while it reduced the levels of ABA continuously at the time of rooting and then increased gradually. Inclusively, our study suggests that IBA 750 mg/L is efficient for the rooting of Magnolia biondii Pamp cuttings, as it enhanced the process of antioxidant enzyme activities, endogenous hormones levels and reduced the time of root formation which is evident from the anatomical observations.
Magnolia biondii Pamp é uma importante espécie de árvore ornamental muito cultivada e utilizada como porta-enxerto na propagação de diferentes variedades de Magnolia. Nos estudos atuais, hormônios anatômicos, fisiológicos e endógenos foram estudados para verificar o efeito do AIB na dose de 750 mg / L no enraizamento adventício e fornecer suporte teórico e técnico para a propagação de M. biondii Pamp por meio de estacas. Duas mil estacas foram preparadas e divididas em dois grupos, ou seja, tratadas com AIB e controle de água. Para a avaliação das atividades das enzimas antioxidantes e dos níveis de hormônios endógenos, as amostras foram coletadas no dia do plantio e a cada 5 dias, enquanto as demais etapas foram realizadas em laboratório de acordo com os protocolos e os devidos cuidados. Para as observações anatômicas, as amostras foram coletadas no 13º, 15º e 17º dias para estacas tratadas com AIB e no 21º, 23º e 25º dias para o controle. As amostras coletadas foram preservadas em solução FAA, e outras observações foram realizadas em laboratório. Observações anatômicas mostraram a necessidade de 13 dias para a diferenciação dos primórdios radiculares até o aparecimento de raízes adventícias jovens em estacas tratadas com AIB e de 21 dias para o desenvolvimento dos primórdios no controle. As atividades das enzimas antioxidantes envolvidas nas ROS foram significativamente maiores nas estacas tratadas com AIB em comparação com o controle. A POD apresentou pico no 13º dia antes da emergência das raízes nas estacas tratadas com AIB, enquanto no 21º dia apresentou pico no controle. A PPO teve pico no 21º dia nas estacas tratadas com AIB e no 29º dia no controle. A SOD apresentou pico no 17º dia nas estacas tratadas com AIB e no 25º dia no controle. A aplicação exógena de AIB aumentou os níveis endógenos de IAA e GA3 em relação ao controle, enquanto reduziu os níveis de ABA continuamente no momento do enraizamento e, em seguida, aumentou gradativamente. Inclusive, nosso estudo sugere que o AIB na dose de 750 mg / L é eficiente para o enraizamento de estacas de M. biondii Pamp, visto que potencializou o processo de atividades de enzimas antioxidantes e os níveis de hormônios endógenos, além de reduzir o tempo de formação de raízes, o que fica evidente nas observações anatômicas.
Subject(s)
Magnolia/growth & development , HormonesABSTRACT
Abstract Magnolia biondii Pamp is an important ornamental tree species widely grown and used as a rootstock in the propagation of different Magnolia varieties. In the current studies, anatomical, physiological and endogenous hormones were studied to check the effect of IBA 750 mg/L on the adventitious rooting and to provide theoretical and technical support for the propagation of Magnolia biondii Pamp through stem cuttings. Two thousand stem cuttings were prepared and divided into two groups i.e., IBA treated cuttings and water control. For the evaluation of antioxidant enzyme activities, and endogenous hormones levels, samples were collected on the day of planting and each 5th day and further steps were carried out in the laboratory according to the protocols and proper precautions. For the anatomical observations, samples were collected on the 13th, 15th, and 17th day for IBA treated cuttings while 21st, 23rd, and 25th day for control. Collected samples were preserved in the FAA solution and further observations were carried out in the laboratory. Anatomical observations showed that it took 13 days for the differentiation of root primordia to the appearance of young adventitious roots in IBA treated cuttings, while it took 21 days to develop primordia in the control. Antioxidant enzyme activities involved in ROS were significantly higher in the IBA treated cuttings compared to control. POD showed a peak on the 13th day before the emergence of roots in IBA treated cuttings while it showed a peak on the 21st day in the control. PPO showed a peak on the 21st day in the IBA treated cuttings while it showed a peak on the 29th day in the control. SOD showed a peak on the 17th day in IBA treated cuttings, while it showed a peak on the 25th day in the control. Exogenous application of IBA enhanced the endogenous IAA and GA3 levels compared to CK, while it reduced the levels of ABA continuously at the time of rooting and then increased gradually. Inclusively, our study suggests that IBA 750 mg/L is efficient for the rooting of Magnolia biondii Pamp cuttings, as it enhanced the process of antioxidant enzyme activities, endogenous hormones levels and reduced the time of root formation which is evident from the anatomical observations.
Resumo Magnolia biondii Pamp é uma importante espécie de árvore ornamental muito cultivada e utilizada como porta-enxerto na propagação de diferentes variedades de Magnolia. Nos estudos atuais, hormônios anatômicos, fisiológicos e endógenos foram estudados para verificar o efeito do AIB na dose de 750 mg / L no enraizamento adventício e fornecer suporte teórico e técnico para a propagação de M. biondii Pamp por meio de estacas. Duas mil estacas foram preparadas e divididas em dois grupos, ou seja, tratadas com AIB e controle de água. Para a avaliação das atividades das enzimas antioxidantes e dos níveis de hormônios endógenos, as amostras foram coletadas no dia do plantio e a cada 5 dias, enquanto as demais etapas foram realizadas em laboratório de acordo com os protocolos e os devidos cuidados. Para as observações anatômicas, as amostras foram coletadas no 13º, 15º e 17º dias para estacas tratadas com AIB e no 21º, 23º e 25º dias para o controle. As amostras coletadas foram preservadas em solução FAA, e outras observações foram realizadas em laboratório. Observações anatômicas mostraram a necessidade de 13 dias para a diferenciação dos primórdios radiculares até o aparecimento de raízes adventícias jovens em estacas tratadas com AIB e de 21 dias para o desenvolvimento dos primórdios no controle. As atividades das enzimas antioxidantes envolvidas nas ROS foram significativamente maiores nas estacas tratadas com AIB em comparação com o controle. A POD apresentou pico no 13º dia antes da emergência das raízes nas estacas tratadas com AIB, enquanto no 21º dia apresentou pico no controle. A PPO teve pico no 21º dia nas estacas tratadas com AIB e no 29º dia no controle. A SOD apresentou pico no 17º dia nas estacas tratadas com AIB e no 25º dia no controle. A aplicação exógena de AIB aumentou os níveis endógenos de IAA e GA3 em relação ao controle, enquanto reduziu os níveis de ABA continuamente no momento do enraizamento e, em seguida, aumentou gradativamente. Inclusive, nosso estudo sugere que o AIB na dose de 750 mg / L é eficiente para o enraizamento de estacas de M. biondii Pamp, visto que potencializou o processo de atividades de enzimas antioxidantes e os níveis de hormônios endógenos, além de reduzir o tempo de formação de raízes, o que fica evidente nas observações anatômicas.
ABSTRACT
The study aimed to explore the effects of inoculation of Rhizophagus intraradices on the biomass, effective component content, and endogenous hormone content of Salvia miltiorrhiza through pot experiments. The number of leaves, plant height, dry weight of aboveground and underground parts, branch number, root number, root length, root diameter, and other biomass were mea-sured by weighing and counting methods. The content of salvianolic acid B, caffeic acid, rosmarinic acid, tanshinone Ⅰ, tanshinone Ⅱ_A, cryptotanshinone, and other effective components was determined by ultra-high performance liquid chromatography. The content of ABA and GA_3 was determined by triple quadrupole mass spectrometry. The correlations between biomass and effective components and between effective components and plant hormones ABA and GA_3 were analyzed. The results showed that R. intraradices significan-tly increased the aboveground dry weight, leaf number, and root number of S. miltiorrhiza by 0.24-0.65 times, respectively. The content of salvianolic acid B and rosmarinic acid in the aboveground part and the content of salvianolic acid B, caffeic acid, rosmarinic acid, tanshinone Ⅰ, and tanshinone Ⅱ_A in the underground part were significantly increased by 0.44-1.78 times, respectively. R. intraradices infection significantly increased the GA_3/ABA values of aboveground and underground parts by 3.82 and 76.47 times, respectively. The correlation analysis showed that caffeic acid, the effective component of the aboveground part, was significantly positively correlated with plant height, tanshinone Ⅱ_A, the effective component of the underground part, was significantly positively correlated with biomass root number, cryptotanshinone, and dry weight, while rosmarinic acid was significantly negatively correlated with dry weight. There were significant positive correlations between cryptotanshinone and ABA, tanshinone Ⅱ_A and ABA and GA_3, and caffeic acid and GA_3. In conclusion, R. intraradices can promote the accumulation of biomass and secondary metabolites of S. miltiorrhiza and regulate the balance between plant hormones ABA and GA_3, thereby promoting the growth of S. miltiorrhiza.
Subject(s)
Salvia miltiorrhiza/chemistry , Plant Growth Regulators/analysis , Plant Roots/chemistryABSTRACT
ObjectiveTo study the effects of foliar spraying of two kinds of compound rhizosphere growth-promoting agents on the growth and physiological characteristics of Angelicae Sinensis Radix （ASR）， as well as the pharmacodynamic components， in order to lay a foundation for providing functional microbial agents for ecological cultivation of ASR. MethodThe compound growth-promoting agents T1 （Pseudomonas CBS5， CBS7 and CBSB） and T2 （Bacillus 5C1， 5C5 and 5C7） with the concentration of 1×108 CFU·mL-1 were sprayed on the leaf surface of the field， and the sterile potato glucose broth medium was used as the control （CK）. The plant growth indexes of ASR were measured by conventional methods， the photosynthetic physiological indexes of ASR were measured by portable photosynthetic measurement system， the enzyme activities of plants and microorganisms were measured by kit method， and the endogenous hormone levels were analyzed by ultra-performance liquid chromatography tandem mass spectrometry. The contents of ferulic acid， senkyunolide I， coniferyl ferulate， senkyunolide A and Z-ligustilide were determined by high performance liquid chromatography. ResultCompared with CK， the two compound inoculants could promote the growth of ASR and increase the biomass， increase the leaf net photosynthetic rate， stomatal conductance， intercellular CO2 concentration， transpiration rate， increase catalase， peroxidase， superoxide dismutase， polyamine oxidase， diamine oxidase and polyphenol oxidase enzyme activities， increase endogenous jasmonic acid， cytokinin and gibberellin levels in plants， increase the contents of ferulic acid， senkyunolide A and Z-ligustilide， reduce the contents of malondialdehyde and abscisic acid， and reduce the incidence of root rot. ConclusionFoliar spraying of two kinds of rhizosphere compound growth-promoting agents can promote the growth， photosynthesis and stress resistance of ASR， and can improve the quality of ASR in different degrees. Comprehensive analysis shows that T1 treatment is better than T2 treatment in the growth-promoting and quality-enhancing of ASR.
ABSTRACT
Schisandra sphenanthera is dioecious and only the fruits of female plants can be used as medicine and food. It is of great significance for the cultivation and production of S. sphenanthera to explore the differences between male and female plants at the non-flowering stage and develop the identification markers at non-flowering or seedling stage. In this study, the transcriptome of male and female leaves of S. sphenanthera at the non-flowering stage was sequenced by Illumina high-throughput sequencing technology and analyzed based on bioinformatics. A total of 236 682 transcripts were assembled by Trinity software and 171 588 were chosen as unigenes. Finally, 1 525 differentially expressed genes(DEGs) were identified, with 458 up-regulated and 1 067 down-regulated in female lea-ves. The down-regulated genes mainly involve photosynthesis, photosynthesis-antenna protein, carbon fixation in photosynthetic or-ganisms, and other pathways. Real-time quantitative PCR(qPCR) identified two genes between male and female leaves and one of them was a HVA22-like gene related to floral organ development and abscisic acid(ABA). Enzyme linked immunosorbent assay(ELISA) was applied to determine the content of ABA, auxin, gibberellin, and zeatin riboside(ZR) in leaves of S. sphenanthera. The results showed that the content of ABA and ZR in male leaves was significantly higher than that in female leaves. The involvement of down-regulated genes in female leaves in the photosynthesis pathway and the significant differences in the content of endogenous hormones between male and female leaves lay a scientific basis for analyzing the factors affecting sex differentiation of S. sphenanthera.
Subject(s)
Abscisic Acid , Gene Expression Profiling , Gene Expression Regulation, Plant , Plant Leaves/genetics , RNA-Seq , Schisandra , TranscriptomeABSTRACT
Objective:To carry out germplasm resource evaluation and new variety breeding of <italic>Murraya paniculata</italic> and improve the germplasm quality， so as to ensure the demand， yield and quality of medicinal materials. Method:Following resource investigation and collection， 17 traits of 107 <italic>M. paniculata</italic> germplasm samples， like plant type， basal diameter， leaf shape， leaf length， and leaf width were determined and then subjected to principal component analysis and factor analysis for screening the principal component factors. Nine primary traits were selected as variables for further cluster analysis using Ward's method and Euclidean distance. According to the characteristics of medicinal parts， the core germplasms were screened out. Then the contents of auxin， zeatin， zeatin nucleoside， isopentenyl adenine， isopentenyl adenine riboside， dihydrozeatin， and dihydrozeatinriboside in the leaves were measured by ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS）， followed by their correlation analysis with agronomic trait. Result:The variation coefficients of petiole length， branching number， and basal diameter were large. The nine main factors could be classified into four categories， with a contribution rate of 72.822%. The cluster analysis with Ward's method and Euclidean distance showed that 107 germplasm samples were clustered into six clusters and 61 core germplasms were identified. Such traits as leaf length， leaf width， petiole length， leaf surface， and petiole color were found to play an important role in the classification of <italic>M. paniculata</italic> germplasms. The content of zeatin nucleoside exhibited significant positive correlations with leaf length （<italic>P</italic><0.01）， petiole length （<italic>P</italic><0.01）， and leaf width （<italic>P</italic><0.05）. Conclusion:These results have laid the foundation for further selection and breeding of <italic>M. paniculata</italic> new varieties.
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Objective To establish a convenient, reliable and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of two kinds of endogenous hormones (testosterone, cortisol) in the saliva of healthy human. Methods A positive ion mode was performed on the Agilent 6410A mass spectrometer. Carbamazepine was used as the internal standard. The saliva samples were centrifuged at 21 912.8×g for 10 min with OMGEA NANOSEP 10K ultrafiltration tubes, and chromatographic separation was achieved on an Agilent ZORBAX SB-C18 column with isocratic elution. The mobile phase was acetonitrile∶0.1% formic acid awter solution (60∶40); flow rate was 0.3 mL/min; injection volume was 10 μL; column temperature was 25 ℃; and the analysis period of each sample was 3 min. Results Testosterone and cortisol had good linear relationship with correlation coefficient (r) being both 0.990. Inter-and intra-day precision was both less than 15%. Conclusion This method can be used to determine the contents of testosterone and cortisol in saliva, which provides a reference for rapid and simple detection of hormone levels in vivo.
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Objective: To study the Methods: of endophytic fungi to break the dormancy of Acanthopanax senticosus seeds and the changes of endogenous hormones and enzymes. Methods: The seeds of A. senticosus were stimulated with endophytic fungi, then treated under thermophilic stratification. The content of endogenous hormones such as GA3, ABA, IAA, IBA, and SA, in the seeds of A. senticosus were tested by HPLC. And the changes of its in vivo enzymes (CAT, SOD, POD, and MDA) activity were tested. Results: Five strains of endophytic fungi from A. senticosus apparently promoted the germination of seeds. In the process of fluctuating temperature stratification, the content of GA3, IAA, IBA, and SA were increased, at the same time, the content of ABA was reduced. The activity of POD and MDA enzymes was significantly reduced, and the enzyme activities of CAT and SOD increased obviously. Conclusion: It is suggested that endophytes have significant effect on the content of seed hormones and enzymes, in addition endophytes could promote the initiation of seed germination.
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Objective: To investigate the effect of exogenous nitric oxide (NO) on breaking the dormancy of Acanthopanax senticosus seeds and the changes in endogenous hormones and enzymes,and provide a basis for breaking the dormancy as well as artificial cultivation of A. senticosus seeds.Method: Different concentrations (1,5,10,20 mmol·L-1) of sodium nitroprusside (NO donor) were used to treat the A. senticosus seeds, and then thermophilic stratification was conducted. The content changes of endogenous hormones such as gibberellin (GA3), abscisic acid (ABA),indolo acetic acid (IAA),indolo butyric acid (IBA) and salicylic acid (SA) at different stratification time (0, 30, 50, 80, 100,130 d) were tested by high performance liquid chromatography (HPLC). The activity change of its in vivo enzymes[catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and malondialdehyde (MDA)] were tested by enzyme-labeled instrument.Result: In the seed germination process of A. senticosus,the contents of GA3,IAA,IBA,and SA were increased gradually,while the content of ABA was reduced gradually. The enzyme activities of POD and MDA were significantly reduced,and the enzyme activities of CAT and SOD were increased obviously. Exogenous NO could increase the seed germination rate and shorten the seed germination time. The effect of 20 mmol·L-1 sodium nitroprusside showed the most obvious effect and 10 mmol·L-1 SNP showed the weakest effect in promoting seed germination,showing an obvious "V" shape for changes.Conclusion: Sodium nitroprusside could promote the seed germination effect of A. senticosus, probably by increasing the content of hormones and enzyme in the stage of seed germination and improving the contents of endogenous NO during germination.
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To reveal the effect of plant growth regulator GA₃ and DPC on the active components and its possible mechanism of Lonicera japonica. GA and DPC were applied at the stage of flower bud differentiation, and the content of active ingredients was measured by LC-MS-MS, the content of endogenous hormones were measured by ELISA, and the expression of key enzyme enes expression was determined by qRT-PCR. The level of endogenous hormone GA₃, IAA, ZR, DHZR and iPA in the GA treatment group was significantly improved, the expression of C4H1, C4H2, 4CL1 and HQT2 were also significantly increased. The content of chlorogenic acid, luteolin, luteoloside, isoquercetin and caffeic acid increased significantly. Spraying DPC did not affect or inhibit the accumulation of active components of L. japonica. Spraying GA can increase the content of endogenous GA₃, thus enhance the expression of C4H1, C4H2, 4CL1 and HQT2, and then increase the content of chlorogenic acid and luteolin.
Subject(s)
Chlorogenic Acid , Lonicera , Plant Growth Regulators , Tandem Mass SpectrometryABSTRACT
Objective In order to study the dynamic variation of endogenous hormones in Paris polyphylla var. yunnanensis inoculated by different foreign arbuscular mycorrhizal fungi species. Methods With sterilized soil as growth substrate, the fresh seeds of P. polyphylla var. yunnanensis were co-cultured with 28 arbuscular mycorrhizal (AM) fungi species under the condition of pot culture at room temperature. The rhizomes and fibrils of P. polyphylla var. yunnanensis were collected, and the contents of endogenous hormone zygotic nucleotides (ZR), gibberellin (GA), indole acetic acid (IAA) and abscisic acid (ABA) were determined by HPLC, then content and proportion of endogenous hormones were analyzed, respectively. Results The results showed that it could increase the mycorrhizal colonization rate and seedling rate of P. polyphylla var. yunnanensis inoculated by different foreign AM fungi species. The content change of ZR, GA, IAA and ABA appears different in the rhizomes and seedling fibrils of P. polyphylla var. yunnanensis after the inoculation of AM fungi species, but it has preferences. Compared with the control group (CK), the content of ZR and GA rose obviously in the rhizomes and seedling fibrils of P. polyphylla var. yunnanensis after the inoculation of AM fungi species, the content of ABA reduced and the content of IAA did not obvious change. The ABA/ZR proportion, ABA/GA proportion and ABA/IAA proportion reduced obviously in the rhizomes of P. polyphylla var. yunnanensis after the inoculation of AM fungi species but there was different change in seedling fibrils of P. polyphylla var. yunnanensis. Conclusion In conclusion, it is inferred that the AM fungi can promote the survival rate of the P. polyphylla var. yunnanensis seedlings, and the different AM fungi strains affect the content of endogenous hormones of P. polyphylla var. yunnanensis.
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To investigate the dynamic changes of endogenous hormones contents in Salvia miltiorrhiza after the inoculation of Glomus versiforme(GV).The contents of endogenous hormones of ABA, ZR, GA, IAA and MeJA by ELISA were measured. Infection rata of GV reached plateau of 90% at the 90th d of inoculation; fresh weight of overground part and leaf number were significantly higher in GV group, with 2.7 and 1.96 fold than that of control; contents of all endogenous hormones apart from ABA in over- and under-ground part were markedly lower (P<0.05) in GV group at the 75th and 90th d, respectively, with 63% to 75% and 45% to 81% of that in control, and were significantly higher (P<0.05) in both over- and under-ground part in GV group at the 105th d, with 1.4 to 1.7 fold higher than that of control; content of ABA in underground part increased significantly at 60th, 75th and 105th d. Arbuscular mycorrhizal fungi could promote the growth of S. miltiorrhiza, and affect the dynamic changes of endogenous hormones contents in different infection periods.
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Hormones are the main materials which regulate the plant growth and the secondary metabolites formation of plant. The effects of hormones on cell biomass and metabolites content and the application progress of honrmones in plant cell suspension culture is reviewed. It covers the influence of exogenous hormones’ categories, concentration and proportion on cell biomass and metabolites content in plant cell suspension culture, the developmental course of the endogenous hormones determination, the changes of endogenous hormones content and its effects on the cell biomass and metabolites content, the relation of interaction between exogenous hormones and endogenous hormones, the study on the effect of new hormone varieties on the culture.
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Objective To establish a regenerative system in vitro and determine the endogenous hormones of Zanthoxylum dissitum. Methods Lamina of Z. dissitum was used as the explant. Callus adventitious bud differentiation was carried out by culturing on MS with different hormones. At the same time,in the course of callus induction,four endogenous hormone of GA3,IAA,ABA,and ZR were determined by ELISA. Results The optimum medium for adventitious bud regeneration of Z. dissitum was MS+6-BA 0.5 mg/L+NAA 0.1 mg/L,and the highest induction rate could reach to 61%. During the process of the differentiation of explants callus,the contents of ZR and IAA were correspondingly higher than other endogenous hormones,the content of ABA was always kept in low level during this process,the content of GA3 was kept in the trend of upgrading during the earlier differentiation period. Conclusion It could be considered that ZR and IAA should be the critical factors in the bud induction. According to the results,a proper adding of GA3in the culture medium could improve the differentiation rate of the adventitious bud. On the other side,ABA might be the negative regulation factor.